EA037533B1 - 6,7,8,9-TETRAHYDRO-3H-PYRAZOLO[4,3-f]ISOQUINOLINE DERIVATIVES USEFUL IN THE TREATMENT OF CANCER - Google Patents

Abstract

The invention relates to compounds of Formula (ID)wherein R1, R14, Q and Y are as defined in the claims, to pharmaceutically acceptable salts thereof and to the use thereof in the prevention or treatment of cancer in a warm-blooded animal.

Description

The present description relates to certain indazole compounds and their pharmaceutically acceptable salts, which selectively inhibit the function of the estrogen receptor and have anti-cancer activity. The present disclosure also relates to the use of said indazole compounds and their pharmaceutically acceptable salts in methods of treating a human or animal body, for example, in the prevention or treatment of cancer. The present description also relates to the methods and intermediates involved in the preparation of these indazole compounds, and to pharmaceutical compositions containing them.

Estrogen receptor alpha (ERa, ESR1, NR3A) and estrogen receptor beta (ERe, ESR2, NR3b) are steroid hormone receptors that are members of a large family of nuclear receptors. Organized similarly to all nuclear receptors, ERa consists of six functional domains (named A to F) (Dahlman-Wright, et al., Pharmacol. Rev., 2006, 58: 773-781), and is classified as a ligand dependent transcription factor, because after binding to a specific ligand (female sex steroid hormone, which is 17b-estradiol (E2)), the complex binds to genomic sequences called estrogen receptor elements (ERE) and interacts with coregulators to modulate the transcription of target genes. The ERa gene is located at 6q25.1 and encodes the 595AA protein, and multiple isoforms can be generated by alternative splicing and translation initiation sites. In addition to the DNA-binding domain (domain C) and the ligand-binding domain (domain E), the receptor contains an N-terminal domain (A / B), a hinge domain (D) that connects domains C and E, and a C-terminal extension (domain F). While domains C and E from ERa and ERe are fairly conserved (96 and 55% amino acid sequence identity, respectively), conservatism of domains A / B, D, and F is low (less than 30% amino acid sequence identity). Both receptors are involved in the regulation and development of the female reproductive tract and, in addition, have functions in the central nervous system, cardiovascular system and in bone metabolism. Genomic ER action occurs in the cell nucleus when the receptor binds to the ERE directly (direct activation or classical pathway) or indirectly (indirect activation or nonclassical pathway). In the absence of a ligand, ERs bind to heat shock proteins, Hsp90 and Hsp70, and the associated chaperone mechanism stabilizes the ligand binding domain (LBD), making it accessible to the ligand. The ligand-attached ER is detached from the heat shock proteins, resulting in a conformational change in the receptor that allows dimerization, DNA binding, interaction with coactivators or corepressors, and modulation of target gene expression. In the nonclassical pathway, AP-1 and Sp-1 are alternative DNA regulatory sequences used by both isoforms of the receptor to modulate gene expression. In this example, ER does not interact with DNA directly, but through binding to other transcription factors associated with DNA, for example c-Jun or c-Fos (Kushner et al., Pure Applied Chemistry 2003, 75: 1757-1769). The exact mechanism by which ER affects gene transcription is poorly understood, but appears to be mediated by numerous nuclear factors that are recruited by a DNA-bound receptor. The recruitment of coregulators is mainly mediated by two protein surfaces, AF2 and AF1, which are located in the E-domain and A / B-domain, respectively. AF1 is regulated by growth factors, and its activity depends on the cell environment and promoter, while AF2 activity is completely dependent on ligand binding. Although the two domains can act independently, maximum ER transcriptional activity is achieved through synergistic interactions through 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 the rapid effects of ER in tissues following E2 administration at times considered too short for genomic action. It is still unclear whether the receptors responsible for the rapid action of estrogen are the same nuclear ER or other steroid receptors associated with the G-protein (Warner, et al., Steroids 2006 21: 91-95), however, a growing the number of pathways induced by E2, eg the MAPK / ERK pathway and activation of endothelial nitric oxide synthase, and the PI3K / Akt pathway. In addition to ligand-dependent pathways, ERa has been shown to have ligand-independent activity mediated by AF-1, which is associated with stimulation of MAPK by mediated growth factor signaling, such as insulin-like growth factor 1 (IGF-1) and epidermal growth factor ( EGF). AF-1 activity depends on Ser118 phosphorylation, and an example of cross-interactions between ER and growth factor-mediated signaling 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: 14911494).

It has been shown that a large number of structurally different compounds bind to ER. Some compounds, such as the endogenous E2 ligand, act as receptor agonists, while others competitively inhibit E2 binding and act as receptor antagonists. Such compounds can be divided into 2 classes depending on their functional effects. Selective estrogen receptor modulators (SERMs) such as tamoxifen are characterized by the ability to act as

- 1 037533 both agonists and antagonists of the receptor, depending on the environment of the cell and the promoter, as well as on the target ER isoform. For example, tamoxifen acts as an antagonist in breast tissue, but acts as a partial agonist in bone, cardiovascular system, and uterus. All SERMs appear to act as AF2 antagonists and gain their partial agonist properties via AF1. Members of the second group, of which the fulvestrant is an example, are classified as complete antagonists and are capable of blocking estrogen activity by completely inhibiting the AF1 and AF2 domains by inducing a unique change in the conformation of the ligand binding domain (LBD) upon binding to the compound, which leads to the complete elimination of the interaction between helix 12 and the rest of the LBD, blocking cofactor recruitment (Wakeling, et al., Cancer Res. 1991, 51: 3867-3873; Pike, et al., Structure, 2001, 9: 145-153).

In the presence of E2, intracellular ERa levels are reduced via the ubiquitin proteasome (Ub / 26S) pathway. Ligand-attached polyubiquitinylation of ERa is catalyzed by at least three enzymes; ubiquitin activated by the ubiquitin-activating enzyme E1 is conjugated by E2 to lysine residues via an isopeptide bond via E3, which is an ubiquitin ligase, and the polyubiquitinylated ERa is then directed to the proteasome for cleavage. Although the regulation of ER-dependent transcription and proteasome-mediated cleavage of the ER are associated (Lonard, et al., Mol. Cell, 2000, 5: 939-948), transcription itself is not required to cleave ERa, and the assembly of the transcription initiation complex is sufficient to to make ERa a target for cleavage by nuclear proteasomes. This E2-induced cleavage process is believed to be necessary to ensure its ability to rapidly activate transcription in response to the need for cell proliferation, differentiation and metabolism (Stenoien, et al., Mol. Cell Biol. 2001, 21: 4404-4412). Fulvestrant is also classified as selective estrogen receptor suppressor (SERD), a subclass of antagonists that can also induce rapid suppression of ERa along the 26S proteasome pathway. In contrast, SERMs such as tamoxifen can increase ERa levels, although the effect on transcription is similar. watched in the case of SERD.

Approximately 70% of breast cancers are characterized by the expression of ER and / or progesterone receptors, indicating that the growth of such tumor cells is hormone dependent. It is believed that in the case of other cancers such as ovarian and endometrial cancers, growth also depends on ERa-mediated signaling. In these patients, therapies can suppress ER-mediated signaling either by antagonizing ligand binding to the ER, such as tamoxifen, which is used to treat early and advanced ER-positive breast cancer in both pre- and post-menopausal regimens; either by antagonizing and suppressing the ERa, such as fulvestrant, which is used to treat breast cancer in women that has progressed despite therapy with tamoxifen or aromatase inhibitors; or by blocking estrogen synthesis, such as aromatase inhibitors, which are used to treat early and advanced ER-positive breast cancer. Although these treatments have had an extremely positive impact on breast cancer treatment, a significant number of patients with ER-expressing tumors show resistance to existing de novo ER-based therapies or develop resistance to these treatments over time. Several different mechanisms have been described to explain the emergence of tamoxifen resistance in primary treatment, which mainly involve switching tamoxifen from acting as an antagonist to acting as an agonist, or through the lower affinity of some cofactors that bind to the tamoxifen-ERa complex, which is shifted due to overexpression of these cofactors, or through the formation of secondary sites that facilitate the interaction of the tamoxifen-ERa complex with cofactors that do not normally bind to the complex. Therefore, resistance may result from the proliferation of cells expressing specific cofactors that stimulate the activity of the tamoxifen-ERa complex. It is also possible that other growth factor mediated signaling pathways directly activate the ER receptor or coactivators to stimulate cell proliferation independently of ligand mediated signaling.

More recently, mutations in ESR1 have been identified as a possible resistance mechanism in tumor samples from patients with metastatic ER-positive cancer and in patient-derived xenograft (PDX) models, with frequency values ranging from 17-25%. Such mutations are predominantly, but not exclusively, in the ligand-binding domain, resulting in mutant functional proteins; examples of amino acid substitutions include Ser463Pro, Val543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn, and Asp538Gly, with substitutions at amino acids 537 and 538 accounting for most of the substitutions currently described. Such mutations have not previously been found in genomes from primary breast cancer specimens described in the Cancer Genome Atlas database. Of 390 ER-positive primary breast cancer specimens, no mutation was found in

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ESR1 (Cancer Genome Atlas Network, 2012, Nature, 490: 61-70). The mutations in the ligand-binding domain are believed to have emerged as a resistance response to aromatase inhibitor endocrine therapies, since in the absence of estradiol such mutant receptors exhibit basal transcriptional activity. The crystal structure of the ER mutated at amino acids 537 and 538 demonstrates that both mutations promoted agonist binding ER conformation by shifting helix 12 to recruit the coactivator and thus mimic agonist-activated wild-type ER ... Published data have demonstrated that endocrine therapies such as tamoxifen and fulvestrant can still bind to the mutant ER and inhibit transcriptional activation to some extent, and that fulvestrant is capable of cleaving the Try537Ser receptor, but more high doses (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)). Therefore, it is possible that certain compounds of formula (ID) or their pharmaceutically acceptable salts (described hereinafter) will be capable of suppressing and antagonizing the 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 occurs, many of them are still characterized by dependence on the activity mediated by the ER, and removal of the receptor by the SERD-based mechanism is the best way to remove the ERa receptor from the cell. Fulvestrant is currently the only SERD approved for clinical use, however, despite its properties with respect to the mechanism of reaction, the pharmacological properties of the drug limit its effectiveness due to the current dose limitation of 500 mg per month, resulting in a renewal of the receptor is less than 50% in patient samples compared to complete receptor suppression observed in experiments with a breast cell line in vitro (Wardell, et al., Biochem. Pharm .. 2011, 82: 122-130).

Therefore, there is a need for new agents that target the ER that have the necessary pharmaceutical properties and a SERD mechanism of action to provide increased therapeutic benefit in early, metastatic cancer and acquired resistance regimens.

Compounds according to the present disclosure have been shown to have potent anti-tumor activity, being useful in suppressing uncontrolled cell proliferation that occurs in cancer. Compounds in accordance with the present description provide an anti-tumor effect, acting as a minimum as a SERD. For example, compounds according to the present disclosure can exhibit antitumor activity due to their ability to suppress estrogen receptor function in a number of different breast cancer cell lines, for example, MCF-7, CAMA-1, BT474 and / or MDA-MB- breast cancer cell lines. 134. Such compounds can be expected to be more useful as therapeutic agents, in particular for the treatment of cancer.

Compounds in accordance with the present disclosure may also exhibit advantageous physical properties (e.g., lower lipophilicity, higher water solubility, higher permeability, lower plasma protein binding and / or greater chemical stability), and / or favorable profiles toxicity (eg, reduced activity against hERG), and / or favorable metabolic or pharmacokinetic profiles compared to other known SERDs. Therefore, such compounds may be particularly useful as therapeutic agents, in particular for the treatment of cancer.

In accordance with one aspect of the present disclosure, there is provided a compound of formula (ID)

Me (ID), where Q is O or NH;

R ¹ is CH2F or CHF2;

R ^{14 is} selected from the group consisting of:

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Preferably ring Y is selected from the group consisting of:

ring Y is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

Preferably, when R ¹ is CH2F.

Preferably, when R ^{14 is} selected from the group consisting of:

A preferred compound of the invention is selected from the group consisting of:

N- (4 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline- 6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine;

6 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl ) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -N - (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine;

3 - ((6S, 8R) -6- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8, 9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropan-1-ol;

N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl ) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine;

(6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3f] | isoquinolin-6-yl) phenyl) -1 - (3-fluoropropyl) azetidin-3-amine;

5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -N - (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine;

N- (4 - ((6S, 8R) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4, 3f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine;

N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f ] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine;

(6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -6- (4- (1- (3-fluoropropyl) azetidin-3-yloxy) -2methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3, 6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol;

N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9- tetrahydro3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine;

5-fluoro-6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline-6- yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline -6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine;

N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6- yl) 3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine;

(6S, 8R) -7- (2,2-difluoroethyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl6,7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

3 - ((6S, 8R) -6- (2,6-difluoro-4- (1- (3-fluoropropyl) azetidin-3-ylamino) phenyl) -8-methyl-8,9-dihydro-3H- 4 037533 pyρazolo [4,3-f] isoquinoline-7 (6H) -yl) -2-fluoro-2-methylpρopan-1-ol;

6 - ((6S, 8R) -7- (2,2-difluoro-ethyl) -8-methyl-6,7,8,9-tetρahydρo-3H-pyρazolo [4,3-f] isoquinolin-6-yl) - N (1 - (3-fluoropropyl) azetidin-3-yl) pyridine-3-amine;

(6S, 8R) -7- (2,2-difluoroρethyl) -6- (5 - ((1- (3-fluoroρnρopyl) azetidin-3-yl) oxy) pyρidin-2-yl) -8-methyl6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

1- (3-fluoroρpρopyl) -N- (3-methoxy-4 - ((6S, 8R) -8-methyl-7- (2,2,2-tρfluoroρethyl) -6,7,8,9-tetρahydρo 3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine;

5-fluoroρ-N- (1- (3-fluoroρpρopyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-tρfluoroρethyl) -6,7, 8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine;

2,2-difluoroρ-3 - ((6S, 8R) -6- (3-fluoroρ-5 - ((1- (3-fluoroρpρopyl) azetidin-3-yl) amino) pyρidin-2-yl) -8- methyl3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol;

6 - ((6S, 8R) -7- (2,2-difluoro-ethyl) -8-methyl-6,7,8,9-tetρahydρo-3H-pyρazolo [4,3-f] isoquinolin-6-yl) - 5φτορ-Ν- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

2,2-difluoroρ-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoroρpρopyl) azetidin-3-yl) oxy) pyρidin-2-yl) -8-methyl-3, 6,8,9 tetρahydρo-7H-pyρazolo [4,3-f] isoquinolin-7-yl) pρopan-1-ol;

2,2-difluoroρ-3 - ((6S, 8R) -6- (3-fluoroρ-5 - ((1- (3-fluoroρpρopyl) azetidin-3-yl) oxy) pyρidin-2-yl) -8- methyl3,6,8,9-tetρahydρo-7H-pyρazolo [4,3-f] isoquinolin-7-yl) pρopan-1-ol and

2,2-difluoroρ-3 - ((6S, 8R) -6- (4 - ((1- (3-fluoroρpρopyl) azetidin-3-yl) amino) -2-methoxyphenyl) -8-methyl 3,6,8 , 9-tetρahydρo-7H-pyρazolo [4,3-f] isoquinolin-7-yl) pρopan-1-ol.

Preferably, when the compound of the invention is N- (1- (3-fluoroρnρopyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-tρfluoroρethyl) - 6,7,8,9-tetρahydρo-3H-pyρazolo [4,3f] isoquinolin-6-yl) pyρidin-3-amine:

ό

SSP '

ΗΝ Υ or a pharmaceutically acceptable salt thereof.

In particular, it is preferred when the compound of the invention is N- (1- (3-fluoroρpρopyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2- tρfluoroρethyl) -6,7,8,9-tetρahydρo-3Нpyρazolo [4,3-f] isoquinolin-6-yl) pyρidin-3-amine:

In accordance with another aspect of the present disclosure, the use of a compound of N- (1- (3-fluoroρpρopyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2, 2-tρfluoroρethyl) 6,7,8,9-tetρahydρo-3H-pyρazolo [4,3-f] isoquinolin-6-yl) pyρidin-3-amine, or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer in a warm-blooded animal. Preferably contemplated is the use of the above compound or a pharmaceutically acceptable salt thereof for the prevention or treatment of breast cancer or gynecological cancer.

This document also describes pharmaceutical compositions that contain a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable excipient.

This document also describes a compound of formula (ID), or a pharmaceutically acceptable salt thereof, for use as a medicament.

Also described herein are combinations of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, with another antineoplastic agent for use in the treatment of cancer.

Additional aspects of the present description will become apparent to a person skilled in the art upon reading this description.

In one embodiment, a compound of formula (ID) as defined above is provided.

In one embodiment, a pharmaceutically acceptable salt of a compound of formula (ID) is provided.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein Q is NH.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein Q is O.

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In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein R ¹ is CH2F.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein Q is O and R ¹ is CH2F.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein Q is NH and R ¹ is CH2F.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein R ^{14 is} selected from the group consisting of:

In one embodiment, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, wherein ring Y is selected from the group consisting of:

In one embodiment, a compound of formula (ID) is provided, wherein the compound is selected from the group consisting of:

N- (4 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

6 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3- f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

3 - ((6S, 8R) -6- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8, 9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropan-1-ol;

N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

(6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3-f] isoquinolin-6-yl) phenyl) -1 - (3-fluoropropyl) azetidine-3-amine and

5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -N - (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine;

or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID) is provided, wherein the compound is selected from the group consisting of:

N- (4 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

6 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3- f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

3 - ((6S, 8R) -6- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8, 9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropan-1-ol;

N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

(6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2methoxyphenyl) -8-methyl-6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline;

N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidine-3-amine;

5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine;

N- (4 - ((6S, 8R) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4, 3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

- 6 037533

N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3- £ ] isoquinolin-6-yl) phenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

(6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -6- (4- (1- (3-fluoropropyl) azetidin-3-yloxy) -2methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4.3 - ^] isoquinoline;

2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3, 6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol;

N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9- tetrahydro3H-pyrazolo [4.3- £] isoquinolin-6-yl) pyridin-3-amine;

5-fluoro-6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline- 6-yl) -N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline -6-yl) phenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

N- (4 - ((6S, 8R) -7- (2,2-difluoroetyl) -8-methyl-6,7,8,9-tetraαhydro-3H-nyrazolo [4,3-f] isoquinoline-6- ul) 3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine;

(6S, 8R) -7- (2,2-difluoroethyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl6,7, 8,9-terahydro-3H-pyrazolo [4,3-f] isoquinoline;

3 - ((6S, 8R) -6- (2,6-difluoro-4- (1- (3-fluoropropyl) azetidin-3-ylamino) phenyl) -8-methyl-8,9-dihydro-3Hpyrazolo [4 , 3- £] isoquinoline-7 (6H) -yl) -2-fluoro-2-methylpropan-1-ol;

6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) - N (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine and (6S, 8R) -7- (2,2-difluoroethyl) -6- (5 - ((1- (3-fluoropropyl ) azetidin-3-yl) oxy) pyridin-2-yl) -8-methyl6,7,8,9-terahydro-3H-pyrazolo [4,3-f] isoquinoline;

or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID) is provided, wherein the compound is selected from the group consisting of:

1- (3-fluoro-propyl) -N- (3-methoxy-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro- 3Hpyrazolo [4.3- £] isoquinolin-6-yl) phenyl) azetidin-3-amine;

5-fluoro-N- (1- (3-fluoro-propyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7, 8.9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridine-3-amine;

2,2-difluoro-3 - ((6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoro-propyl) azetidin-3-yl) amino) pyridin-2-yl) -8- methyl3,6,8,9-tetrahydro-7H-pyrazolo [4.3- £] isoquinolin-7-yl) propan-1-ol;

6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) - 5Φτορ-Ν- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine;

2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1 - (3-fluoropropyl) azetidin-3-yl) oxy) pyridin-2-yl) -8-methyl-3, 6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol;

2,2-difluoro-3 - ((6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoro-propyl) azetidin-3-yl) oxy) pyridin-2-yl) -8- methyl3,6,8,9-tetrahydro-7H-pyrazolo [4.3- £] isoquinolin-7-yl) propan-1-ol;

2,2-difluoro-3 - ((6S, 8R) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) -2-methoxyphenyl) -8-methyl 3,6,8 , 9-tetrahydro-7H-pyrazolo [4,3- £] isoquinolin-7-yl) propan-1-ol;

or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID) is provided which is N- (1- (3-fluoro-propyl) azetidin-3-yl) -6- (8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3Hpyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine, or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID) is provided which is N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2 , 2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine, or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID) is provided which is N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6R, 8R) -8-methyl-7- (2,2 , 2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine, or a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided, wherein the compound is selected from the examples herein. An additional feature is any of the embodiments described herein, provided that any of the specific examples are excluded in isolation. An additional feature is any of the embodiments described herein, with the proviso that any one or more compounds selected from the above list of exemplary compounds in accordance with the present disclosure are separately excluded.

Additionally, for the avoidance of ambiguity, the use in the formulas herein denotes a point of attachment between different groups.

Compounds of formula (ID) have two or more chiral centers, and it will be understood that such a compound of formula (ID) can be prepared, isolated and / or supplied in the presence, or in the absence, or in addition to one or more of other possible enantiomeric and / or diastereomeric isomers of the compound of formula (ID) in any relative proportion. Getting enantiomerically

- 7,037,533 enriched / enantiomerically pure and / or diastereomerically enriched / diastereomerically pure compounds can be carried out using standard organic chemistry techniques well known in the art, for example, by synthesis from enantiomerically enriched or enantiomerically pure starting materials, using the corresponding enantiomerically enriched or enantiomerically pure catalyst during synthesis and / or by separation of a racemic or partially enriched mixture of stereoisomers, for example, by means of chiral chromatography.

For pharmaceutical applications, it may be preferred to prepare a compound of formula (ID) or a pharmaceutically acceptable salt thereof without the presence of other stereoisomeric forms in large amounts.

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

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

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

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

In an additional embodiment, a composition is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, optionally together with one or more other stereoisomeric forms of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, wherein the compound of formula (ID) or a pharmaceutically acceptable salt thereof present in the composition in an enantiomeric excess (% ee) of> 90%.

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

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

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

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

In additional embodiments of the aforementioned composition,% ee and% de may have any combination of the values listed below.

% it is <%, and% de is> 80%.

% of it is <5% and% de is> 90%.

% of it is <5% and% de is> 95%.

% of it is <5% and% de is> 98%.

% is> 95% and% de is> 95%.

% is> 98% and% de is> 98%.

% is> 99% and% de is> 99%.

In an additional embodiment, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable excipient.

In one embodiment, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable excipient, optionally further comprising one or more other stereoisomeric forms of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, wherein the compound of formula (ID) or a pharmaceutically acceptable salt thereof are present in the composition in an enantiomeric excess (% ee) of> 90%.

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

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

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

In one embodiment, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable excipient, optionally further comprising one or more other stereoisomeric forms of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, wherein the compound formula (ID) or a pharmaceutically acceptable salt thereof are present in the composition in a diastereomeric excess (% de) of> 90%.

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

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

- 8 037533

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

In one embodiment, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable excipient, optionally further comprising one or more other stereoisomeric forms of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, wherein the compound formula (ID) or a pharmaceutically acceptable salt thereof is present in the composition in an enantiomeric excess (% ee) of> 0% and in a diastereomeric excess (% de) of> 90%.

In additional embodiments of the aforementioned pharmaceutical composition,% ee and% de may have any combination of the values listed below.

% is> 95% and% de is> 95%.

% is> 98% and% de is> 98%.

% is> 99% and% de is> 99%.

Compounds of formula (ID) and their pharmaceutically acceptable salts may be prepared, used or supplied in amorphous form, crystalline form or semi-crystalline form, and any specified compound of formula (ID) or a pharmaceutically acceptable salt thereof may be capable of forming more than one crystalline / polymorphic form , including hydrated forms (for example, hemihydrate, monohydrate, dihydrate, trihydrate or hydrate of another stoichiometric composition) and / or solvated forms. It should be understood that the present description covers all such solid forms of the compound of formula (ID) and its pharmaceutically acceptable salts.

In additional embodiments, the implementation provides a compound of formula (ID) obtainable using the methods described later in this document in the Examples section.

The present invention is intended to include all isotopes of atoms found in the compounds of the present invention. It should be understood that isotopes contemplate those atoms that have the same atomic number, but different mass numbers. For example, the isotopes of hydrogen include tritium and deuterium. Carbon isotopes provide ¹³ C and ¹⁴ C. Nitrogen isotopes provide ¹⁵ N.

A suitable pharmaceutically acceptable salt of a compound of formula (ID) is, for example, an acid addition salt. A suitable pharmaceutically acceptable salt of a compound of formula (ID) may be, for example, an acid addition salt of a compound of formula (ID), for example an acid addition salt formed with an inorganic or organic acid such as acetic acid, adipic acid, benzenesulfonic acid, benzoic acid, cinnamic acid, citric acid, D, L-lactic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, hydrochloric acid, L-tartaric acid, maleic acid, malic acid, malonic acid, methanesulfonic acid, napisylic acid, phosphoric acid, saccharin, succinic acid, sulfuric acid, p-toluenesulfonic acid, toluenesulfonic acid, or trifluoroacetic acid.

An additional suitable pharmaceutically acceptable salt of a compound of formula (ID) is, for example, a salt formed in a human or animal body after administration of a compound of formula (ID) to said human or animal body.

The compound of formula (ID), or a pharmaceutically acceptable salt thereof, can be prepared as a co-crystalline solid form. It should be understood that a pharmaceutically acceptable co-crystal of a compound of formula (ID), or pharmaceutically acceptable salts thereof, forms an aspect of the present disclosure.

Specific solid forms described herein have substantially the same X-ray powder diffraction patterns as the X-ray powder diffraction patterns shown in the figures, and are characterized by different 2-theta angle values shown in the tables incorporated herein. It will be understood that the values of the 2-theta angle of the powder X-ray diffraction pattern may vary slightly from one device to another or from one sample to another, and therefore the values given should not be taken as absolute.

It is known that it is possible to obtain a powder X-ray diffraction pattern that has one or more measurement errors depending on the measurement conditions (such as the equipment or device used). In particular, it is generally known that the intensity values in the X-ray powder diffraction pattern can fluctuate depending on the measurement conditions. Therefore, it should be understood that solid forms in accordance with the present disclosure are not limited to crystals that have X-ray powder diffraction patterns that are identical to the X-ray powder diffraction patterns shown in the Figures, and any crystals having X-ray powder diffraction patterns are substantially the same as shown in the Figures. are within the scope of the present description. An expert in the field of powder X-ray diffraction is able to conclude that the X-ray powder diffraction patterns are largely identical.

It will be appreciated by those skilled in the X-ray powder diffraction field that the relative intensity of the peaks can be influenced, for example, by grains larger than 30 μm and with a range of values.

- 9 037533 aspect ratios that can affect the analysis of samples. One of ordinary skill in the art will also understand that the position of the reflections can be influenced by the exact height at which the sample is in the diffractometer and the zero calibration of the diffractometer. The flatness of the sample surface can also have little effect. Therefore, the presented data of diffraction patterns should not be taken as absolute values. (Jenkins, R. & Snyder, RL 'Introduction to X-Ray Powder Diffractometry, John Wiley & Sons 1996; Bunn, CW (1948), Chemical Crystallography, Clarendon Press, London; Klug, HP & Alexander, LE (1974), X-Ray Diffraction Procedures).

Typically, the diffraction angle measurement error in the powder X-ray diffraction pattern is about ± 0.2 ° of the 2-theta angle, and this degree of measurement uncertainty should be taken into account when considering the X-ray powder diffraction patterns in the figures when interpreting the data contained in the tables included in this document. In addition, it should be understood that the intensity values can fluctuate depending on the experimental conditions and the method of obtaining the sample (preferred orientation).

In this description, it was initially found that the compound N- (1- (3-fluoro-propyl) azetidine-3yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridin-3-amine (hereinafter, compound X) is in the form of an amorphous solid. Useful crystalline polymorphic forms of the compound were subsequently prepared using the conditions described in the experimental section.

Therefore, in a further aspect of the present disclosure, polymorphic form A of compound X is provided. This polymorphic form can be characterized in that it provides at least one of the following 2θ angle values measured using CuKa radiation: 15.5, 18.6 and 24, 6 °.

Polymorphic Form A of Compound X is characterized in that it provides an X-ray powder diffraction pattern essentially as shown in FIG. one.

The ten powder X-ray diffraction peaks for a given polymorphic form [2-theta (2θ) angle, intensity (%)] are as follows: 21.1 (100%), 20.8 (54.3%), 14.6 (41, 9%), 18.6 (41.6%), 12.3 (38.9%), 15.5 (34.1%), 24.6 (31.3%), 15.8 (30, 6%), 13.4 (23.2%) and 19.0 ° (21.7%).

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately 15.5 ° 2-theta.

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately a 2-theta angle of 18.6 °.

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately a 2-theta angle of 24.6 °.

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least two characteristic peaks at approximately 2-theta angles of 15.5 ° and 18.6 °.

In accordance with the present disclosure, polymorphic form A of compound X is provided, which is characterized by an X-ray powder diffraction pattern with characteristic peaks at approximately 2-theta angle values of 21.1, 20.8, 14.6, 18.6, 12.3, 15, 5, 24.6, 15.8, 13.4 and 19.0 °.

In accordance with the present disclosure, polymorphic form A of compound X is provided which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. one.

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta angle of 15.5 ± 0.2 ° 2-theta.

In accordance with the present disclosure, polymorphic form A of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta angle value of 18.6 ± 0.2 ° 2-theta angle.

In accordance with the present disclosure, polymorphic form A of compound X is provided, which is characterized by an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta angles of 15.5 and 18.6 °, where these values may deviate by ± 0.2 ° angle 2-theta.

In accordance with the present description, polymorphic form A of compound X is provided, which is characterized by an X-ray powder diffraction pattern with specific peaks at 2theta angle values of 21.1, 20.8, 14.6, 18.6, 12.3, 15.5, 24 , 6, 15.8, 13.4 and 19.0 °, where the indicated values may deviate by ± 2 ° of the 2-theta angle.

In a further aspect of the present disclosure, polymorphic form E of compound X is provided. This polymorphic form can be characterized in that it provides at least

- 10 037533 one of the following 2θ angle values measured using CuKa radiation: 14.8 °, 16.2 ° and 17.9 °.

Polymorphic Form E of Compound X is characterized by providing an X-ray powder diffraction pattern substantially as shown in FIG. eight.

The ten powder X-ray diffraction peaks for a given polymorphic form [2-theta (2θ) angle, intensity (%)] are as follows: 17.9 ° (100%), 14.8 ° (67.1%), 20.9 ° (60.1%), 23.1 ° (55.4%), 16.2 ° (49.3%), 20.0 ° (35.6%), 18.2 ° (32.9%) , 12.3 ° (30.4%), 22.2 ° (19.0%) and 13.9 ° (18.9%).

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately a 2-theta angle of 17.9 °.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately a 2-theta angle of 14.8 °.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at approximately a 2-theta angle of 17.9 °.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least two characteristic peaks at approximately 2-theta angles of 17.9 ° and 14.8 °.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with characteristic peaks at approximately 2-theta angles of 17.9, 14.8, 20.9, 23.1, 16.2, 20, 0, 18.2, 12.3, 22.2 and 13.9 °.

In accordance with the present disclosure, there is provided a polymorphic form E of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. eight.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta angle of 17.9 ± 0.2 ° 2-theta.

In accordance with the present disclosure, polymorph E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta angle of 14.8 ± 0.2 ° 2-theta.

In accordance with the present disclosure, a polymorphic form E of compound X is provided which is characterized by an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta angles of 17.9 ° and 14.8 °, where these values may deviate by ± 0.2 ° angle 2-theta.

In accordance with the present description, polymorph E of compound X is provided, which is characterized by an X-ray powder diffraction pattern with characteristic peaks at 2-theta angle values of 17.9, 14.8, 20.9, 23.1, 16.2, 20.0 , 18.2, 12.3, 22.2 and 13.9 °, where the indicated values may deviate by ± 0.2 ° of the 2-theta angle.

It should be understood that a suitable pharmaceutically acceptable prodrug of a compound of formula (ID) also forms an aspect of the present invention. Accordingly, compounds according to the present disclosure can be administered in the form of a prodrug, which is a compound that is cleaved in the human or animal body to release the compound disclosed herein. A prodrug can be used to alter the physical properties and / or pharmacokinetic properties of a compound as described herein. A prodrug can be formed if a compound according to the present disclosure contains a suitable group or substituent to which a property-modifying group can be attached. Examples of prodrugs include in vivo cleavable ester or amide derivatives of the compound of formula (ID).

Accordingly, one aspect of the present disclosure includes those compounds of formula (ID) as defined herein above if formed by organic synthesis and if formed in a human or animal body by cleavage of a prodrug of a compound of formula (ID). Accordingly, the present disclosure includes those compounds of formula (ID) that are obtained by organic synthesis methods, as well as those compounds that are formed in the human or animal body by metabolism of the precursor compound, thus the compound of formula (ID) may be a compound obtained by synthesis, and a compound obtained by metabolism.

A suitable pharmaceutically acceptable prodrug of a compound of formula (ID) is one which, according to sound medical judgment, is suitable for administration to a human or animal body without undesirable pharmacological activities and without undue toxicity.

- 11 037533

The different forms of the prodrug are described, for example, in the following documents:

a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, etal.

(Academic Press, 1985);

b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985);

c) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 Design and Application of Pro-drugs, by H.

Bundgaard p. 113-191 (1991);

d) H. Bundgaard, Advanced Drug Delivery Reviews 8, 1-38 (1992);

e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988);

f) N. Kakeya, et al., Chern, Pharm. Bull. 32, 692 (1984);

g) T. Higuchi and V. Stella, Pro-Drugs as Novel Delivery Systems, A.C.S.

Symposium Series, Volume 14 and

h) E. Roche (editor), Bioreversible Carriers in Drug Design, Pergamon Press, 1987.

The effects of a compound of formula (ID) in vivo may be due in part to one or more metabolites that are generated in a human or animal body following administration of a compound of formula (ID). As indicated above in this document, the effects of the compound of formula (ID) in vivo can also be manifested through the metabolism of the precursor compound (prodrug).

For the avoidance of ambiguity, it should be understood that if in the present description a group is designated as defined above in this document or as defined in this document, then the specified group includes the first and widest definition, as well as all alternative definitions for this group without exception.

Another aspect of the present disclosure provides a method for preparing a compound of formula (ID), or a pharmaceutically acceptable salt thereof. A suitable method is illustrated by the following exemplary method variations. The required starting materials can be obtained using standard organic chemistry procedures. The preparation of such starting materials is described in conjunction with the following illustrative method variations and within the scope of the accompanying examples. Alternatively, the required starting materials can be obtained using procedures similar to those illustrated, which are within the ordinary skill of a person skilled in the art of organic chemistry.

Compounds of formula (ID) can be prepared, for example, by:

a) esterification of a suitable aryl or heteroaryl compound of formula (II) wherein L is, for example, halogen (such as iodine) or a trifluoromethanesulfonyl (triflate) group or boronic acid or ester with an alcohol of formula (III) with using a suitable metal catalyst (eg RockPhos 3rd generation pre-catalyst) in a suitable solvent (such as toluene or DME) in the presence of a suitable base (such as cesium carbonate) and at a suitable temperature (such as 90-120 ° C); removing the protecting group (PG) in formula (II), such as THP, using acidic conditions (such as anhydrous HCl in 1,4-dioxane) at a suitable temperature (such as 10-30 ° C):

b) amination of a suitable aryl or heteroaryl compound of formula (II), where L, for example, is a halogen (such as iodine) or trifluoromethylsulfonyloxy (triflate) group, with an amine of formula (IV) using a suitable metal catalyst (for example, BrettPhos or RuPhos and Pd ₂ (dba) ₃ ) in a suitable solvent (e.g. 1,4-dioxane) in the presence of a suitable base (e.g. cesium carbonate, sodium t-butoxide or LiHMDS) at a suitable temperature (such as 90-130 ° C) ; removing the protecting group (PG) such as THP using acidic conditions (such as anhydrous HCl in 1,4-dioxane) at a suitable temperature (such as 10-30 ° C):

- 12 037533

c) alkylating a suitable phenol or hydroxylheteroaryl compound of formula (V) with an alcohol of formula (III) via a Mitsunobu reaction using appropriate reagents (such as triphenylphosphine and diisopropyl (E) -diazen-1,2-dicarboxylate) in a suitable solvent (such as THF); removing a protecting group (PG) such as THP in formula (VII) using acidic conditions (such as anhydrous HCl in 1,4-dioxane) at a suitable temperature (such as 10-30 ° C):

d) alkylating a suitable aniline or heteroarylamine or phenol or hydroxylheteroaryl compound of formula (VI) with a compound of formula (VII) wherein LG is a leaving group (such as a halide or mesylate) using weak bases (e.g. DIPEA) in a suitable solvent (such as DMF or MeCN); removing the protecting group (PG) such as THP using acidic conditions (such as anhydrous HCl in 1,4-dioxane) at a suitable temperature (such as 10-30 ° C):

f) alkylating amines of formula (VIII) with a suitable alkylating group of formula (IX) (where LG can be a halide such as bromide, iodide or chloride, or can be another suitable leaving group such as mesylate) in a suitable solvent ( such as DMF) in the presence of a suitable base (such as DIPEA) at a suitable temperature (such as 10-30 ° C):

Compounds of formula (II) wherein R ^{6 is} not hydrogen can be prepared, for example, from compounds of formula (X) by oxidation with a suitable reagent (e.g. bis (trifluoroacetoxy) iodobenzene) and treatment with an organometallic reagent (e.g. bromomethyl magnesium, if R ⁶ is methyl) in a suitable solvent (eg THF) at low temperature (typically -80 to -60 ° C).

- 13 037533

Compounds of formula (X) can be prepared by, for example, reacting aniline of formula (XI) with reagents suitable for providing indazole formation, such as inorganic nitrite (such as sodium nitrite), in an organic acid (such as propionic acid) at low temperature (typically -20 to 0 ° C) or alternatively with an acid anhydride (such as acetic anhydride) in the presence of a suitable base (such as potassium acetate) together with an organic nitrite (such as isopentyl nitrite) optionally in the presence of a crown ether (such as 18-crown-6) in a suitable solvent (such as chloroform) at a suitable temperature (such as 70 ° C).

Compounds of formula (XI) can be prepared by reacting a compound of formula (XII) with a compound of formula (XIII) under conditions known in the art as suitable for Pictet-Spengler reactions, such as in the presence of an acid (such as acetic acid) and in a suitable solvent (eg toluene or water) and at a suitable temperature (such as 60-100 ° C).

Compounds of formula (XII) can be prepared by functional group interconversions known in the art, for example, by amination reactions of halides of formula (XIV) from aryl halides (such as bromide) using a protected amine (such as diphenylmethanimine) in the presence of a suitable catalyst and ligand (such as bis (dibenzylideneacetone) palladium (0) and rac-2,2'-bis- (diphenylphosphino) -1,1'-binaphthyl) in the presence of a suitable base (such as sodium t-butoxide) in a suitable solvent (such as toluene) at a suitable temperature (such as 80-100 ° C).

Compounds of formula (XIV) can be prepared using:

a) reacting a compound of formula (XV) with an aldehyde of formula (XVI) in a suitable solvent (eg THF) in the presence of a suitable reducing agent (such as sodium triacetoxyborohydride) and at a suitable temperature (such as 20-30 ° C);

b) (i) reacting a compound of formula (XV) with an acid of formula (XVII) under standard conditions to form an amide bond (e.g., 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)), then (ii) reducing the formed 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);

c) reacting a compound of formula (XV) with a compound of formula (XVIII), wherein LG is a suitable leaving group (e.g., a halogen atom (such as bromine or chlorine) or a triflate), in the presence of a suitable base (such as diisopropylethylamine) in a suitable solvent (eg DCM or dioxane) and at a suitable temperature (such as 20-85 ° C).

- 14 037533

Compounds of formula (XV) can be prepared by several methods known in the art, primarily as methods for synthesizing chiral amines.

a) Ring opening of sulfamidates of formula (XIX) according to the scheme shown below.

Stage 1. Alkylation, for example, n-butyllithium / THP / -78-0 ° C.

Stage 2. Removal of protective groups, for example anhydrous HCl in MeOH / DCM, rt.

b) Phase transfer alkylation in the presence of a chiral catalyst (such as (1S, 2S, 4S, 5R) -2 ((R) - (αlyloxy) (quinolin-4-yl) methyl) -1 - (anthracene-9-ylmethyl) - 5-vinylquinuclidine-1-th bromide) with subsequent processing of the functional group:

Bg

Stage 1

Stage 2 ^Bg

ΝΗ

R ^{7 R}

Step 1. Alkylation, eg chiral catalyst, toluene / KOH, 0 ° C.

Stage 2. Interconversion of the functional group.

Compounds of formula (XII) can be directly prepared using:

a) reacting a compound of formula (XX) with an aldehyde of formula (XVI) in a suitable solvent (eg THF) in the presence of a suitable reducing agent (such as sodium triacetoxyborohydride) and at a suitable temperature (such as 20-30 ° C);

b) (i) reacting a compound of formula (XX) with an acid of formula (XVII) under standard conditions to form an amide bond (e.g. 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)), then (ii) reducing the formed 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);

c) reacting a compound of formula (XX) with a compound of formula (XIII), wherein LG is a suitable leaving group (e.g., a halogen atom (such as bromine or chlorine) or a triflate), in the presence of a suitable base (such as diisopropylethylamine) in a suitable solvent (e.g. DCM or dioxane) and at a suitable temperature (such as 20-85 ° C):

Compounds of formula (XX) can be prepared using a sequence of reactions starting with protected 3-bromo-2-methyl-aniline as shown below.

Stage 1

Stage 2 Stage 3

R (XX)

H;

(xix)

Stage 1. Alkylation, for example n-butyllithium / THF / from -78 ° C to rt.

Stage 2. Removal of protecting groups intended for the amine, for example, anhydrous HCl in MeOH / DCM, etc.

Stage 3. Removal of protecting groups intended for aniline, for example heating under reflux in hydroxylamine.

- 15 037533

Compounds of formula (V) can be prepared by a Pikte-Spengler reaction cyclizing a compound of formula (XIII) containing a boronic acid ester as described above to give a compound of formula (XXI). A compound of formula (XXI) can be oxidized to a compound of formula (V) using a suitable oxidizing agent (such as hydrogen peroxide) in the presence of a suitable base (such as sodium hydroxide) in a suitable solvent (such as THF).

Compounds of formula (VI) wherein Q is NH can be prepared by a sequence of reactions comprising cyclizing a nitro-containing compound of formula (XIII) in a Pictet-Spengler reaction as described above to give a compound of formula (XXII). A compound of formula (XXII) can be reduced to a compound of formula (VI) using suitable nitro reduction conditions (such as hydrogenation) in the presence of a suitable catalyst (such as platinum dioxide) in a suitable solvent (such as methanol).

(XXII)

Compounds of formula (VIII) wherein Q is O can be prepared from aryl halides of formula (II) and tert-butyl 3-hydroxyazetidine-1-carboxylate using a suitable metal catalyst (such as RockPhos 3rd generation precatalyst) in a suitable solvent (such as toluene or DME) in the presence of a suitable base (such as cesium carbonate) at a suitable temperature (such as 90-120 ° C); The Boc protecting group can be subsequently removed using an acid (such as trifluoroacetic acid) in a suitable solvent (such as DCM). Compounds of formula (VIII) (Q is O) can also be prepared from a compound of formula (V) under conditions known in the art as suitable for Mitsunobu reactions using appropriate reagents (such as triphenylphosphine and diisopropyl- (E) -diazen -1,2 dicarboxylate) with tert-butyl 3-hydroxyazetidine-1-carboxylate in a suitable solvent (such as THF).

Compounds of formula (VIII) wherein Q is NH can be prepared from aryl halides of formula (II) and tert-butyl 3-aminoazetidine-1-carboxylate using a suitable metal catalyst (e.g. RuPhos or BrettPhos and Pd ₂ (dba) ₃ ) in a suitable solvent (eg 1,4-dioxane) in the presence of a suitable base (eg cesium carbonate, sodium t-butoxide or LiHMDS) at a suitable temperature (such as 90-130 ° C); the Boc protecting group can be subsequently removed using an acid (such as trifluoroacetic acid) in a suitable solvent (such as DCM).

Compounds of formula (III) can be prepared by:

a) carrying out an alkylation reaction between 3-hydroxyazetidine and compounds of formula (IX), where LG is, for example, halogen or another leaving group (such as a mesyl group), in the presence of a suitable base such as cesium carbonate in a suitable solvent such as acetonitrile, at a suitable temperature such as 120 ° C and in a suitable container such as a sealed tube;

b) carrying out a reductive amination reaction between 3-hydroxyazetidine and aldehyde or ketone compounds of formula (XXIII) in the presence of a suitable reducing reagent such as sodium triacetoxyborohydride in a suitable solvent such as DCM at a suitable temperature such as 10- 30 ° C.

y- R ¹ o = ^ R ¹² (XXIII)

Compounds of formula (IV) can be prepared by:

a) (i) carrying out an alkylation reaction between a compound of formula (XXIV), where PG is

- 16 037533 represents a protecting group, for example Boc, and compounds of formula (IX) where LG is, for example, halogen or another leaving group such as mesylate in the presence of a suitable base such as DIPEA in a suitable solvent such as 1,4-dioxane, at a suitable temperature such as 10-30 ° C; (ii) removing the protecting group under suitable conditions such as acidic conditions to remove Boc;

(XXIV)

b) (i) carrying out a reductive amination reaction between compounds of formula (XXIV) and aldehyde or ketone compounds of formula (XXIII) in the presence of a suitable reducing reagent such as sodium triacetoxyborohydride in a suitable solvent such as DCM at a suitable temperature such as 10-30 ° C; (ii) removing the protecting group under suitable conditions such as acidic conditions to remove Boc.

It should be understood that other variations of the process steps are also possible in the process variations described above.

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

A suitable protecting group for an amino group or an alkylamino group is, for example, an acyl group, for example an alkanoyl group such as an acetyl, alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or tert-butoxycarbonyl group, an arylmethoxycarbonyl group, for example a benzyloxycarbonyl, or an aroyl group, for example a benzoyl group. The deprotection conditions for the above-mentioned protecting groups inevitably vary depending on the choice of the protecting group. For example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group can be removed, for example, by hydrolysis using a suitable base such as an alkali metal hydroxide such as lithium or sodium hydroxide. Alternatively, an alkoxycarbonyl group such as a t-butoxycarbonyl group can be removed, for example, by treatment with a suitable acid such as hydrochloric, sulfuric, formic, phosphoric or trifluoroacetic acid, and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group can 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 the primary amino group is, for example, a phthaloyl group, which can 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 diarylalkylsilane, such as TBDMS or TBDPS. The deprotection conditions for the above protecting groups will inevitably vary depending on the choice of the protecting group. For example, an acyl group such as an alkanoyl or aroyl group can be removed, for example, by hydrolysis using a suitable base such as an alkali metal hydroxide, such as lithium or sodium hydroxide. Alternatively, an arylmethyl group such as a benzyl group can be removed, for example, by hydrogenation over a catalyst such as palladium on carbon.

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

Protecting groups can be removed at any convenient stage of the synthesis using conventional techniques well known in the field of chemistry.

Some of the intermediates defined in this document are new, and they are presented as additional features of the present description.

- 17 037533

In one embodiment, a compound of formula (XXV) or a salt thereof is provided:

L

(xxv) where L is bromine, chlorine, iodine, or trifluoromethanesulfonyl.

In an additional embodiment, L is bromine.

Biological analyzes

The following assays were used to measure the effects of compounds as described herein.

Binding analysis ERa.

The ability of compounds to bind to the isolated estrogen receptor alpha ligand binding domain (LBD ER-alpha (GST)) was evaluated in competitive assays using LanthaScreen ™ time resolved fluorescence resonance energy transfer (TR-FRET) endpoint detection. A suitable fluorophore (Fluormone ES2, ThermoFisher, product code P2645) and recombinant ligand binding domain of human estrogen receptor alpha, residues 307-554 (expressed and purified in situ) were used to measure compound binding by TR-FRET LanthaScreen ™ endpoint. The principle of the assay is that LBD ER-alpha (GST) is added to a fluorescent ligand to form a receptor-fluorophore complex. Terbium-labeled anti-GST antibodies (product code PV3551) are used to indirectly label the receptor by binding to its GST-tag, and competitive binding is monitored by the ability of the test compound to displace fluorescent ligand, resulting in a loss of TRFRET signal between the Tb-labeled anti-GST antibody and the tag. The assay was performed as follows with all reagents added using a BioRAPTR FRD microfluidic workstation from Beckman Coulter.

1. Using an acoustic dispenser, dispense 120 nL of test compound into black 384-well assay plates with a small well volume.

2. Prepare 1x LBD ER-alpha / Tb-labeled GST Ab in ES2 screening buffer and incubate for 15 min.

3. Dispense 6 μl of reagent containing 1x LBD AR / Tb-labeled anti-GST Ab into each well of the assay plate, then dispense 6 μl 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. Provide excitation at 337 nm and measure the fluorescence signal for each well at 490 and 520 nm using BMG PhERaSTAR.

Compound was dosed directly from a compound source microplate containing serially diluted compound (4 wells containing 10, 0.1, 1 mM, and 10 nM final compound, respectively) into an assay microplate using a Labcyte Echo 550. Echo 550 is is a liquid dispenser that uses acoustic technology to directly transfer compound solutions in DMSO from microplate to microplate, and the system can be programmed to transfer various small volumes, on the order of nL, compound from different wells of the original plate to obtain the required serial dilution of the compound in the assay plate, which is then refilled to normalize the DMSO concentration over the entire dilution range.

A total of 120 nL of compound with DMSO was added to each well, and the compounds were tested in a 12-point concentration response curve format over the final concentration range of the compound, including 10, 2.917, 1.042, 0.2083, 0.1, 0, respectively. , 0292, 0.0104, 0.002083, 0.001, 0.0002917, 0.0001042 and 0.00001 μM. TR-FRET dose response data obtained for each compound was exported to a suitable software package (such as Origin or Genedata) for curve fitting analysis. Competitive binding of ER-alpha was expressed as an IC ₅₀ value. It was determined by calculating the concentration of the compound that was necessary to reduce the binding of the tagged compound to LBD ER-alpha by 50%.

Analysis of ER suppression in MCF-7.

The ability of compounds to suppress the number of estrogen receptor (ER) was assessed in an immunofluorescence assay on cells using the MCF-7 cell line obtained from carcino

- 18 037533 we of the human breast ducts. MCF-7 cells were reconstituted directly from a cryoflacon (approximately 5x106 cells) in an assay medium (Dulbecco's Modified Eagle's Medium (DMEM) without phenol red; Sigma D5921) containing 2 mM L-glutamine and 5% (v / v) fetal charcoal / dextran treated calf whey. Cells were harvested once using a large gauge sterile needle syringe 18Gx1.5 "(1.2x40 mm) and the cell density was measured using a Coulter counter (Beckman). Cells were further diluted in assay media to a density of 3.75x104 cells / ml and added at 40 μl per well to clear bottom 384-well black plates (Costar, # 3712) treated for tissue culture using Thermo Scientific Matrix WellMate or Thermo Multidrop. After seeding the cells, the plates were incubated overnight at 37 ° C, 5% CO ₂ (Liconic carousel incubator). Test data was obtained using a LabCyte Echo ™ Model 555 Mixer Reformator, which is part of an automated production cell (Integrated Echo 2 production cell). Compound stock solutions (10 mM) of the test compounds were used to provide a 384 well dose plate for compound (Labcyte P-05525-CV1). 40 μl of each 10 mM compound stock solution was dispensed into the well of the first quadrant and then serial 1: 100 dilutions were made in DMSO using a Hydra II liquid dispenser (MATRIX UK) to obtain 40 μl of the diluted compound, respectively, in the wells of quadrant 2 (0 , 1 mM), 3 (1 μM) and 4 (0.01 μM). The addition of 40 μL of DMSO to the wells of row P of the original plate allowed DMSO to be normalized over the entire dose range. To dose control wells on the original compound plate, 40 μL of DMSO was added to row O1 and 40 μL of 100 μM fulvestrant in DMSO was added to row O3.

The Echo uses acoustic technology to perform direct microplate-to-microplate transfer of DMSO compound solutions to assay plates. The system can be programmed to multi-step transferring small volumes, down to 2.5 nL, between microplates and thus serially dilute the compound in the assay plate, which is then refilled to normalize the DMSO concentration throughout the dilution range. Compounds were dispensed into cell plates, with the original compound plate prepared as above to give 12 concentration points, with a dose range of 3 μM to 3 pM, each in duplicate, using 3-fold dilutions and one final 10 -fold dilution using the Integrated Echo 2 production cell. Control wells with maximum signal were dosed with DMSO to give a final concentration of 0.3% and control wells with minimum signal were dosed with fulvestrant to give a final concentration of 100 nM, respectively. The plates were additionally incubated for 18-22 h at 37 ° C, 5% CO ₂ , and then fixed by adding 20 μl of 11.1% (v / v) formaldehyde solution (in phosphate-buffered saline (PBS)) to obtain the final formaldehyde concentration 3.7% (v / v). Cells were fixed at room temperature for 20 min and then washed twice with 250 μl PBS / Proclin (PBS with biocidal preservative) using a BioTek plate washer, then 40 μl PBS / Proclin was added to all wells and the plates were stored at 4 ° C. The above fixation method was performed on an Integrated Echo 2 production cell. Immunostaining was performed using an AutoElisa automated production cell. PBS / Proclin was removed from all wells by evacuation and cells were permeabilized with 40 μl PBS containing 0.5% Tween ™ 20 (v / v) for 1 hour at room temperature. Plates were washed three times in 250 μl PBS / 0.05% (v / v) Tween 20 using Proclin (PBST with biocidal preservative) and then 20 μl 1: 1000 rabbit anti-ERa monoclonal antibody solution (SP1) (Thermofisher) was added in PBS / Tween ™ / 3% (w / v) bovine serum albumin. The plates were incubated overnight at 4 ° C (Liconic carousel incubator) and then washed three times in 250 μl PBS / 0.05% (v / v) Tween ™ 20 with Proclin (PBST). The plates were then incubated with goat anti-rabbit IgG with AlexaFluor 594 or goat anti-rabbit immunoglobulins with AlexaFluor 488 (Molecular Probes) at 20 μl / well with 1: 5000 Hoechst solution in PBS / Tween ™ / 3% (w / v .) bovine serum albumin for 1 hour at room temperature. The plates were then washed three times in 250 μl PBS / 0.05% (v / v) Tween ™ 20 with Proclin (PBST with biocidal preservative). 20 μl of PBS was added to each well and the plates were covered with black plate film and stored at 4 ° C. prior to reading. Plates were read using Cellomics Arrayscan, reading the fluorescence level at 594 nm (time point 24 hours) or 488 nm (time point 5 hours) to measure the ERa receptor level in each well. The mean total intensity was normalized to the number of cells to give the total intensity per cell. The data was exported to a suitable software package (such as Origin) for curve fit analysis. The level of suppression of the ERa receptor was expressed as an IC ₅₀ value and was determined by calculating the concentration of the compound that was necessary to obtain a 50% reduction in the mean of the maximum signal of the total intensity.

Were obtained the data shown in table. A (the data below may represent

- 19 037533 is the result of a single experiment or the average of two or more experiments).

Table A

Example IC50 value IC50 value for

to bind ER (nM) suppression ER (nM) ¹ one 3.7 0.10 2 2.8 3.1 3 6.5 0.64 four 6.9 0.34 five 2.2 0.077 6 8.8 0.35 7 8.4 0,4 eight 2.0 0.055 nine 360 > 300 10 eight 0.26 AND twenty 0.094 12 2.4 0.25 (83%) 13 3.5 oh and fourteen 6.6 > 300 fifteen 9.5 0.33 (84%) sixteen 6.9 0.54 17 2.4 0.16 eighteen 29 0.42 nineteen 2.6 0.063 twenty fifteen 0.36 21 2.1 0.098 22 5.7 0.062 23 fifteen 0.15 24 10 0.21 25 4.1 0.33 26 6,7 0.52 27 0.81 0.05 28 5.7 0.065 29 1.6 0.2

- 20 037533

thirty 1.8 0.28 31 13 0.54 32 2.1 0.096 33 1.4 ο, ι 34 1.9 0.22 35 1.5 0.17 36 1.8 0.12 37 7.2 4.1 38 0.69 0.19 39 0.94 0.048 40 1,3 ο, ι 41 2.4 0.17 42 3.8 0.28 43 4.1 0.25 44 9.6 0.91 45 1.6 oh h 46 3.6 0,4 47 1,2 0.41 48 1.5 0.078 49 1.5 0.075 fifty 2.7 0.47 51 2.8 0.68 52 AND 0.21 53 5.7 0.26 54 160 > 300 55 8.2 1.1 56 8.2 0.58 57 five 0.36 58 2.1 0.56 59 7.2 1.6 60 4.7 0.19

61 0.88 0.13 62 one 0.13 63 73 0.31 64 6.1 0.22 65 5.3 one 66 6.2 0.44 67 five 0.27 68 fifteen 0,4 69 8.9 0.55 70 4.3 0.31 71 2 0.17 72 2.9 0.51 73 3.4 0.77

- 21 037533 'Compounds that are active in the ER suppression assay exhibit suppression values> 90% in the assay, unless otherwise indicated, in which case the% suppression is shown in parentheses.

Western blot analysis.

The ability of compounds to suppress the estrogen receptor (ER) was assessed by Western blotting using human breast cancer cell lines (MCF-7 and CAMA-1). Cells were plated in 12-well tissue culture-treated plates at 0.5x10 ⁶ / well in RPMI without phenol red containing 2 mM L-glutamine and 5% (v / v) fetal charcoal-treated calf serum (F6765 , Sigma). The cells were incubated with compounds (100 nM) or a control medium (0.1% DMSO) for 48 h at 37 ° C, 5% CO ₂ , after which they were washed once with PBS and lysed with 80 μl lysis buffer (25 mM Tris / HCl, 3 mM EDTA, 3 mM EGTA, 50 mM NaF, 2 mM sodium orthovanadate, 0.27 M sucrose, 10 mM β-glycerophosphate, 5 mM sodium pyrophosphate, 0.5% TritonX- 100, pH6.8) on ice.

Cells were scraped, sonicated and centrifuged (Protein Assay Kit 500-0116, DC Bio-Rad) prior to protein assay, then samples were adjusted to a protein concentration of 1-2 mg / ml in lysis buffer containing 1x LDS sample buffer (NP0007, Invitrogen) and 1x NuPAGE Sample Recovery (NP0009, Invitrogen). The samples were boiled for 10 minutes at 95 ° C and then frozen at -20 ° C until the need for use.

10-20 μg protein was loaded onto 26-well Criterion gels (BioRad 345-0034). The gels were incubated at 125 V for 1 h 25 min in a rolling buffer (24 mM Tris Base Sigma, 192 mM glycine, 3.5 mM SDS prepared in distilled water). Then, at 30 V for 2 h in a blotting buffer (25 mM Tris, 192 mM glycine, 20% (v / v) methanol, pH 8.3, prepared in distilled water), the gels were transferred to a nitrocellulose membrane. The blotting membrane was stained with Ponceau S (P7170, Sigma) and excised according to the appropriate molecular weight markers.

The membranes were blocked for 1 hour at room temperature in 5% Marvel (w / v) in phosphate buffered saline containing 0.05% Tween ™ 20 (PBS / Tween). The blotting membranes were then incubated with anti-ERa rabbit monoclonal antibody (SP1) (Thermofisher) diluted 1: 1000 at 4 ° C. overnight (with gentle shaking) followed by several washes with PBS / Tween. Secondary antibody to rabbit immunoglobulins conjugated to HRP (7074, CST), diluted to a dilution of 1: 2000, was incubated for 2 h at room temperature (with gentle shaking) followed by several washes with PBS / Tween. All antibodies were prepared in 5% Marvel (w / v) in PBS / Tween.

Immunoblots were developed using Pierce WestDura chemiluminescence reagents (Thermo Scientific 34076) and developed / quantified on a G-box using Syngene software. The ERa receptor suppression value was normalized against the vehicle control (0% suppression) and the 100 nM fulvestrant control (100% suppression) run on the same gel.

Table B shows data obtained for selected examples (data below may represent the result of a single experiment or the average of two or more experiments).

- 22 037533

Table B

Example Western blotting for CAMA1% degr. ER versus Fv Western blotting for MCF7% deg. ER versus Fv one 92 91 2 95 93 3 102 90 four 61 70 five 109 99 6 84 89 7 90 90 eight 100 101 nine 25 -13 sixteen 105 104 17 96 92 eighteen 95 103 nineteen 94 92 21 101 99 25 96 98 26 97 97 27 108 110 28 110 95 29 106 93 thirty 98 96 31 95 103 32 89 89 33 102 98 34 82 85 40 120 98 42 79 92 46 102 105 48 94 93 49 99 98 53 46 81 56 52 61 59 102 104 64 89 86 69 98 95 70 100 102

Analysis using human hepatocytes.

The metabolic stability of compounds in human hepatocytes was assessed using the following protocol.

1. Prepare 10 mM stock solutions of compound and control compounds in an appropriate solvent (DMSO). The incubation medium (L-15 medium) is placed in a water bath at 37 ° C and heated for at least 15 minutes before use.

2. Add 80 μl of acetonitrile to each well of a 96-well deep well plate (block plate).

- 23 037533

3. In a new 96-well plate, dilute 10 mM test compounds and control compounds to 100 µM by combining 198 µl acetonitrile and 2 µl 10 mM stock solution.

4. Remove the vial containing cryopreserved (at temperatures less than -150 ° C) human hepatocytes (10 donor LiverPool ™ human hepatocytes obtained from Celsis IVT. Chicago, IL (Product # S01205)) from storage, making sure the vials remain cryogenic. temperatures until the thawing process begins. Thaw the cells as quickly as possible by placing the vial in a water bath at 37 ° C and gently shaking the vials. The vials should remain in the water bath until all ice crystals have dissolved and are no longer visible. After completion of defrosting, the vial is sprayed with 70% ethanol, the vial is transferred to a microbiological safety box.

5. Open the vial and pour the contents into a 50 ml conical tube containing thawing medium. Place a 50 ml conical tube in a centrifuge and centrifuge at 100 g for 10 minutes. After completion of centrifugation, the thawing medium is removed by evacuation and the hepatocytes are resuspended in a sufficient amount of incubation medium to obtain ~ 1.5x10 ⁶ cells / ml.

6. Using a Cellometer® Vision, cells are counted and the density of viable cells is determined. Cells with low viability (viability <80%) are unacceptable. Cells are diluted with incubation medium to a working cell density of 1.0x10 ⁶ viable cells / ml.

7. Transfer 247.5 μl of hepatocytes to each well of a 96-well cell culture plate. Place the plate on an Eppendorf Thermomixer Comfort plate shaker to heat the hepatocytes for 10 min.

8. Add 2.5 µL of 100 µM test compound or control compounds to the incubation well containing the cells, mix until a homogeneous suspension is achieved after 0.5 min, which, when reached, will define a time point of 0.5 min. After 0.5 min, 20 μl of the incubated mixture is transferred to the wells in the plate to block, followed by vortexing.

9. The plate is incubated on an Eppendorf Thermomixer Comfort plate shaker at 37 ° C at 900 rpm. After 5, 15, 30, 45, 60, 80, 100 and 120 min, the incubation system is stirred and at each time point samples of the incubated mixture with a volume of 20 μl is transferred to the wells in a separate blocking plate, followed by vortexing.

10. The blocking plates are centrifuged for 20 minutes at 4000 rpm, 4 different compounds are collected in one cassette and used for LC / MS / MS analysis.

All calculations were performed using Microsoft Excel. Peak areas were determined from extracted ion chromatograms. The in vitro intrinsic clearance (in vitro Cl _int , in l / min / 10 ⁶ cells) of the parent compound was determined by regression analysis of the Ln curve of the percent disappearance of the parent compound versus time. The in vitro intrinsic clearance (in vitro Clint, in L / min / 10 ⁶ cells) was determined from the slope value using the following equation, and for selected examples its values are shown in Table 1. FROM.

in vitro Clint = kV / Ν where V = incubation volume (0.25 ml); N = number of hepatocytes per well (0.25x10 ⁶ cells).

Physical properties.

logD.

The lipophilicity of a drug is an important physical property that can

- 24 037533 affect a variety of biological and metabolic properties of a compound, such as absorption, distribution, metabolism, excretion and toxicity profiles of the compound. The partition coefficient between 1-octanol and an aqueous buffer, LogDO / W, at pH 7.4 is the most widely used indicator of the lipophilicity of a compound. The current LogDO / W measurement method is based on the traditional shake flask technique, but with the modification of measuring ten compounds simultaneously in mixtures using UPLC with quantitative mass spectrometry (MS) as a method for measuring relative concentrations in octanol and water. The maximum capacity is 379 working connections (48 sets of 10 connections, including three QC connections) per experiment. Two quality control (QC) samples, medium LogD cyclobenzaprine and high LogD nicardipine, are used with all populations to ensure proper quality. An additional QC sample, caffeine, with a low LogD value, was used and randomly allocated to all cycles. The method has been thoroughly tested against previously used shake flask techniques.

Solubility.

In order for an oral compound to reach the site of action, and for an oral compound to be absorbed from the intestine, the compound must be in solution, and therefore compounds that have high intrinsic solubility may be more suitable for pharmaceutical use. The thermodynamic solubility of the test compound is measured under standard conditions. This is a shake flask method that uses 10 mM DMSO based solutions supplied from Compound Managements' liquid storage facility and has a high throughput. The dried compounds are equilibrated in aqueous phosphate buffer (pH 7.4) for 24 h at 25 ° C, then the part with the dissolved compound is separated from the residue. Solutions are analyzed and quantified using UPLC / MS / MS, and QC samples are included in each run to ensure assay quality. Human Plasma Protein Binding Human plasma protein binding is a key factor in controlling the amount of free (unbound) drug available for target binding and therefore plays an important role in the observed drug efficacy in vivo. Therefore, compounds that have a high free fraction (low plasma protein binding) may show increased efficacy relative to compounds with similar levels of activity and exposure. An automated equilibrium dialysis assay in human plasma is performed using a RED (Rapid Equilibrium Dialysis) device and sample processing. The analysis usually lasts from two to three days, including the issuance of results. After dialysis for 18 hours, samples of blood plasma and buffer are prepared for analysis by liquid chromatography and mass spectrometry. Samples are usually tested in one replicate and quantified by LC / MSMS using a 7-point calibration curve in blood plasma. The compounds are combined into plasma aggregates of no more than 10 compounds. Three reference compounds are used in each cycle: propranolol, metoprolol and warfarin. Warfarin was used as a control in each set, and propranolol and metoprolol were randomly placed in each cycle. The Excel macro for internal use is used to prepare files for the robotic station and mass spectrometer and is also used to calculate the content of the unbound fraction (fu%) in blood plasma.

Table D shows LogD, solubility and plasma protein binding data obtained for selected examples (data below may represent the result of a single experiment or the average of two or more experiments).

Table D

Example LogD pH 7.4 Binding to human blood plasma protein (% free) Solubility (μM) one 3.4 8.3 379

- 25 037533

2 2.6 28 217 3 2.5 26 > 941 four 2.7 21 991 five h, h 8.6 297 6 2.4 34 > 947 7 3.5 AND 470 eight 4.4 0.99 90 nine one 21 > 1000 10 2.8 23 634 AND 3.9 2 186 12 3.9 2.7 77 13 four 6.4 78 fourteen 2.6 sixteen 897 fifteen 2.5 34 821 sixteen 2 64 982 17 2.9 23 833 eighteen 2.6 34 910 nineteen 3.7 4.3 152 twenty 2.8 29 730 21 3.1 12 634 22 four 4.6 70 23 3.1 9.5 339 25 2.5 40 964 26 3 22 258 27 3.8 3.8 229 28 3.6 6 150 29 s sixteen 779 thirty s sixteen 754 31 2 44 > 942 32 3.5 6.5 692 33 3.6 5.4 466

- 26 037533

34 3.2 fifteen 471 37 2.6 25 961 38 3.1 13 794 39 3.7 6.8 257 40 3.5 6.4 479 41 4.1 2.2 120 42 2.4 n. and. 712 44 3.2 10 998 45 3.9 6.2 172 46 2.5 31 > 1000 47 2.6 n. and. > 1000 48 3.1 10 582 49 3.8 n. and. 163 fifty 2.1 n. and. 748 51 2.7 28 > 1000 52 3 nineteen 864 53 3.2 AND 691 54 1.4 57 > 1000 55 2.1 n. and. 987 56 3.6 n. and. 377 57 2.9 n. and. 615 58 2.4 n. and. 633 59 2.4 33 971 60 2.5 23 593 61 2.7 29 848 62 2.8 nineteen 784 63 3.2 10 870 64 3.2 fourteen 474 65 2.4 n. and. 790 66 2,3 n. and. 931 67 2.7 nineteen 914

68 2.4 37 561 69 2.9 37 > 1000 70 2.9 24 857 71 3.2 n. and. 666 72 3.3 fourteen 738 73 s 22 925

n. and. = not investigated.

HERG binding assay

Potassium channels encoded by hERG (human heart specific potassium channel gene) are important for normal electrical activity of the heart. Arrhythmia can result from blockage of hERG channels by a heterogeneous group of drugs. This side effect is a common cause of unsatisfactory results in preclinical drug safety trials [Sanguinetti et al., Nature., 2006, 440, 463-469.], And therefore minimizing hERG channel blocking activity can be a relevant property for drug candidate.

The purpose of the hERG binding assay is to evaluate the effects of test compounds on the voltage-gated potassium channel encoded by the human heart specific potassium channel gene (hERG) using a constitutively expressing CHO cell line on a Nanion Syncropatch 384PE automated voltage clamping system. Analysis was performed as follows, with all reagents being used at room temperature unless otherwise indicated.

Reagent preparation includes:

1) IC700 internal solution used for pouring into the bottom of the chip (in mM): 130 KF,

KCl, 1 MgCl ₂ , 10 EGTA and 10 HEPES (all from Sigma-Aldrich; pH 7.2-7.3 using 10 M KOH,

320 mosm) and with the addition of 25 μM escin;

2) external buffer and buffer for cells (in mM): 137 NaCl, 4 KCl, 10 HEPES, 10 D-glucose, 2 CaCl ₂ , 1 MgCl2 (pH 7.4, NaOH);

3) NMDG reference buffer used to establish a stable baseline before adding test compounds (in mM): 80 NaCl, 4 KCl, 2 CaCl ₂ , 1 MgCl ₂ , 60 NMDG Cl, 5 D-glucose monohydrate, 10 HEPES (pH 7 , 4 NaOH 298 mOsm);

4) a contact enhancer used to improve the ability of cells to form contact (in mM): 80 NaCl, 3 KCl, 10 CaCl ₂ , 10 HEPES, I MgCl ₂ (pH 7.4 NaOH).

Cell preparations.

1. When using cell culture, cells must be incubated at 30 ° C for about 4-6 days prior to use. On the day of analysis, cells are harvested using accutase and resuspended in 20 ml of cell buffer to a density of 0.8 to 1e6 ml cells.

2. If ready for analysis cryovials are used, quickly thaw two cryovials at 37 ° C and slowly pipette into 23 ml of external solution.

3. Before starting the analysis, all preparative. cells must be incubated for 15 min in a shaker-incubator for cells set at 10 ° C.

Preparation of compounds.

All compounds were dispensed by acoustic technology in four replicates using Labcyte Echo. A 10 mM stock solution was used to prepare 6 compound stock plates, each at a different concentration, for applying cumulative doses to the cells (0.03167, then 0.1, then 0.3167, 1, 3.167, 10 mM). 90 μl of reference buffer containing 600 nl of compound is added to each well of the original plates to give final compound concentrations of 0.1, 0.39, 1.2, 3.9, 12.5 and 39.6 μM, respectively.

HERG assay (all dispensing steps were performed on a Nanion syncropatch using a liquid dosing setting).

1. Add 40 μl of external buffer to 384 well 4-channel chips with medium resistance and pour the internal buffer into the bottom of the plate.

2. Dispense 20 µl of cells into each well of the chip, followed by 20 µl of contact enhancer.

3. Remove 40 µl of reagent from each well to the washing block, leaving a residual volume of 40 µl.

4. Dispense 40 µl of reference buffer with a step of removing 40 µl after 3 minutes, repeat this step.

5. Dispense 40 μl of compound from plate 1 (0.03167 mM), record in real time with 3 min exposure, then remove 40 μl. This step is repeated for another 5 further compound plates at increasing concentrations to generate a cumulative concentration-response curve for each well of the Syncropatch chip.

The hERG-mediated currents were generated using a voltage drop protocol consisting of intermittently maintaining the voltage at -80 mV, with a step of increasing to 60 mV for 500 ms, followed by a step of decreasing to -40 mV for 500 ms in 15 s increments. The current through the hERG was automatically measured based on the leakage subtraction recording using Nanion software by recording the peak of the hERG trace current at -40 mV in 15 s increments and using the last three such responses for each concentration to construct a concentration-effect curve.

The calculation of the results is carried out using the APC package as part of Genedata. For standard normalization of well data using neutral control well groups and inhibitor control well groups as a reference, the GeneData Assay Analyzer applies the following equation to normalize signal values to the desired signal range:

N (x) = CR + x - <cr> (SR - CR) <sr> - <cr>

where x is the original value of the measured signal from the well;

<cr> is the average of the measured signals from the wells on the plate, which are the central standard (neutral);

<sr> is the average of the measured signals from the wells on the plate representing the scale standard (with inhibitor);

CR is the required value for the central standard (neutral), normalized to the mean;

SR is the required value for the reference scale (with inhibitor), normalized to mean.

Table E shows hERG binding data for selected examples (data below may represent the result of a single experiment or the average of two or more experiments).

Table E

Example IC ₅₀ hERG (μM) Example IC ₅₀ hERG (μM) one 10 41 6.5 2 > 40 42 7.9 3 > 36 45 4.2 four 17 46 > 40 five 13 49 8.2 6 9.1 fifty 13 sixteen > 40 51 9.1 17 22 52 2 eighteen > 38 55 > 40 nineteen 5.4 57 sixteen 21 5.3 58 > 40 25 28 59 > 40 27 7.9 60 AND 28 7.8 61 > 40 29 27 62 > 40 thirty > 33 63 13 31 > 40 64 13 32 fourteen 65 31 33 fourteen 66 26 34 7,7 67 fourteen 37 twenty 68 > 40 38 eighteen 69 24 39 fourteen 70 24 40 4.5 73 12

Permeability.

In order to maximize the absorption of the compound for oral administration, the drug should have a sufficient level of transmembrane transfer, and also avoid excretion by P-glycoprotein. The most widely used system for predicting the absorption of a compound for oral administration is to determine the level of penetration of compounds through the monolayers of the human colon adenocarcinoma cell line Caco-2.

Bidirectional permeability of human Caco-2 from A to B and B to A.

To determine the bidirectional permeability (elimination and capture) of compounds in Caco-2 cells, an automated assay was used, which was performed for 2 h at pH 7.4. Samples were analyzed by LC / MS / MS to evaluate the apparent coefficients of permeability (Papp) of compounds in monolayers of Caco-2 cells, and the results are reported in units of x10 ^-6 cm / s.

The elimination rate (ER) can be determined using the following equation:

ER = Papp (BA) / Papp (AB), where P _app ( _BA ) denotes the apparent permeability coefficient for the direction from the basolateral to the apical side;

P _app ( _AB ) denotes the apparent permeability coefficient for the apical to basolateral direction.

P _app for the passive permeability of the human Caco-2 in the direction from A to B.

To determine the passive permeability of compounds in monolayers of Caco-2 cells, an automated analysis was used, which was carried out for 2 h at pH 6.5 on the apical side and pH 7.4

- 29 037533 on the basolateral side. The Caco-2 AB inhibition assay is performed using chemical inhibition in Caco-2 cells of three major efflux transporters: ABCB1 (P-gp), ABCG2 (BCRP), and ABCC2 (MRP2). Incubation of both the apical and basolateral sides is carried out using a mixture of inhibitors (50 μM quinidine, 20 μM sulfasalazine and 100 μM benzbromarone). Samples were analyzed by LC / MS / MS to evaluate the apparent permeability coefficients (P _app ) of compounds in monolayers of Caco-2 cells, and the results are reported in units of x10 ^-6 cm / s.

Table F shows permeability data obtained for selected examples (data below may represent the result of a single experiment or the average of two or more experiments).

Table F

Example Rarr for bidirectional permeability Saco-2 (x10 ' ⁶ cm / s) Elimination rate for bidirectional permeability Saso-2 Rarr for passive permeability Saco-2 (x10 ' ⁶ cm / s) one 0.9 9.4 n. and. 2 3.9 8.4 n. and. 3 2,3 9.2 n. and. 6 0.9 28 1.8 AND 1.6 1.4 n. and. sixteen 1.6 9,7 1.7 17 4.7 5.5 13.3 nineteen 4.2 2.6 n. and. 21 4.0 1.6 14.1 25 5.0 7.4 5.9 27 5.4 0.9 n. and. 28 1,2 3.8 24 29 12 0.9 24 thirty 2.7 7.8 n. and. 31 0.9 9.4 n. and. 32 3.4 1.5 n. and. 33 2.6 1.5 n. and. 47 eighteen 1.1 n. and. 60 0.7 34 n. and.

64 2.0 5.6 n. and.

n. and. = not investigated.

Anti-tumor efficacy in mice against xenograft representing parental human MCF7.

To determine the effect of Example 17 on the growth of MCF7 xenografts, the following study was performed. Before implantation, MCF7 cells (ATCC) were grown in vitro at exponential phase. Briefly, male SCID mice weighing 18 g or more (Envigo UK) were subcutaneously implanted with estrogen pellets (0.5 mg, 21 days release, from Innovative Research of America) on the back using a renewable anesthetic agent. One day later, mice were inoculated subcutaneously from the left flank with 5 million MCF7 cells obtained as 0.1 ml of cell suspension in a 1: 1 mixture of RPMI (Gibco, Life Technologies) and Matrigel (Corning). When tumors reached ~ 250 mm ³ , mice were randomly assigned to groups of 9 mice (12 mice for vehicle control) and drug treatment started. Compounds were prepared in vehicle (40% tetraethylene glycol (v / v), 7.5% captisol (w / v) in water for injection) and given orally in a volume of 10 ml / kg once a day for 21 days at a dose of 0.5 to 50 mg / kg. Tumors were measured twice weekly and tumor volume was calculated using an elliptical body formula (pi / 6 x width x width x length). Data represent the geometric mean tumor volume relative to tumor volume on the day of distribution in random order. Error bars represent 95% confidence intervals (Graphpad Prism). This study demonstrated that doses of 10 mg / kg and higher resulted in tumor regression (Fig. 12).

Antitumor efficacy in mice against human MCF7 xenograft with Y537S mutation in ESR1.

To determine the effect of Example 17 on the growth of xenografts derived from MCF7 cells genetically engineered to express Y537S ESR1, the following study was performed. MCF7 cells with Y537S in ESR1 are generated by genome editing and express only Y537S ESR1 (Ladd, et al., Oncotarget, 2016, 7: 54120-54136). Briefly, male SCID mice weighing 18 g or more (Envigo UK) were inoculated subcutaneously from the left flank using 5 million MCF7 cells with Y537S in ESR1, obtained as 0.1 ml of cell suspension in a 1: 1 RPMI mixture (Gibco, Life Technologies ) and matrigel (Coining). When tumors reached ~ 250 mm ³ , mice were randomly assigned to groups of 9 mice (12 mice for vehicle control) and drug treatment started. Compounds were prepared in vehicle (40% tetraethylene glycol (v / v), 7.5% captisol (w / v) in water for injection) and given orally at a volume of 10 ml / kg once a day for 22 days at a dose of 0.5 to 50 mg / kg. Tumors were measured twice weekly and tumor volume was calculated using an elliptical body formula (pi / 6x width width length). Data represent the geometric mean of tumor volume relative to tumor volume on the day of distribution in random order. Error bars represent 95% confidence intervals (Graphpad Prism). This study demonstrated that doses of 10 mg / kg and higher resulted in tumor regression (Fig. 13).

Antitumor efficacy in mice against human xenograft CTC174 obtained from a patient with breast cancer and mutation in ESR1.

To determine the effect of Example 17 on the growth of a patient-derived CTC174 xenograft with a mutation in ESR1, female NSG mice were implanted with CTC174 fragments in mammary adipose tissue. CTC174 was obtained from circulating tumor cells isolated from a patient with metastatic ER + breast cancer and shown to contain the D538G mutation in ESR1 at an allele frequency of 0.33 (Ladd, et al., Oncotarget. 2016, 7: 54120-54136 ). Briefly, female NOD / SCID mice after oophorectomy (Cg-Prkdcscid Il2rgtm1Wj1 / SzJ) (NSG) (6-7 weeks old - The Jackson Laboratory) under the influence of renewable anesthesia were implanted with a xenograft fragment of ~ 50 mm CTC174 ³ into the adipose tissue of the third mammary gland. When the tumors reached ~ 200 mm ^{3, the} mice were randomly assigned to groups of 10 mice and drug treatment was started. Compounds were prepared in vehicle (40% tetraethylene glycol (v / v), 7.5% captisol (w / v) in water for injection) and given orally at a volume of 10 ml / kg once a day for 32 days at a dose of 0.8 to 40 mg / kg. Tumors were measured twice weekly and tumor volume was calculated using an elliptical body formula (pi / 6 x width x width x length). Data represent the geometric mean of tumor volume relative to tumor volume on the day of distribution in random order. Error bars represent 95% confidence intervals (Graphpad Prism). This study demonstrated that doses of 10 mg / kg and above provided almost complete suppression of tumor growth (Fig. 14).

Efficacy of a combination of antitumor drugs in a mouse against a human xenograft CTC174 obtained from a patient with breast cancer and a mutation in ESR1.

To determine the effect of Example 17 on the growth of a CTC174 xenograft obtained from a patient with a mutation in ESR1, in combination with palbociclib, a CDK4 / 6 inhibitor, or vistusertib, an mTORC1 / 2 inhibitor (AZD2014), female NSG mice fragments of CTC174 were implanted into the adipose tissue of the mammary gland. CTC174 was obtained from circulating tumor cells isolated from a patient with metastatic ER + breast cancer and shown to contain the D538G mutation in ESR1 at an allele frequency of 0.33 (Ladd, et al., Oncotarget, 2016, 7: 54120-54136 ). Briefly, female NOD / SCID (Cg-Prkdcscid Il2rgtm1Wj1 / SzJ) (NSG) mice (6-7 weeks old - The Jackson Laboratory) under the influence of renewable anesthesia were implanted with a CTC174 xenograft fragment with a volume of ~ 30 mm ³ in adipose tissue of the third mammary gland. When the tumors reached ~ 500 mm ³ , the mice were randomly assigned to groups of 10 mice and started with drug treatment. Example 17 was prepared in a vehicle (40% tetraethylene glycol (v / v), 7.5% captisol (w / v) in water for injection) and was given orally at a volume of 10 ml / kg once a day in within 23 days at a dose of 10 mg / kg. Palbociclib and visusertib were prepared in carrier medium (1% polysorbate 80). Palbociclib was given orally at a volume of 10 ml / kg once daily for 23 days at a dose of 50 mg / kg. Visusertib was given orally at a volume of 10 ml / kg twice a day on a schedule 2 days after 5 for 23 days at a dose of 10 mg / kg. Carrier-treated group was dosed orally with 10 ml / kg

- 31 037533

40% tetraethylene glycol (v / v), 7.5% captisol (w / v) in water for injection once daily for 23 days. Tumors were measured twice weekly and tumor volume was calculated using an elliptical body formula (pi / 6x width x width x length). Data represent the geometric mean of tumor volume relative to tumor volume on the day of distribution in random order. Error bars represent 95% confidence intervals (Graphpad Prism). This study demonstrates that Example 17 in combination with either palbociclib (FIG. 15) or visusertib (AZD2014) (FIG. 16) provided a greater effect than either of the agents administered alone.

According to a further aspect of the present disclosure, there is provided a pharmaceutical composition that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in combination with a pharmaceutically acceptable excipient.

Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents, granulating agents and disintegrating agents, binders, lubricants, preservatives, and antioxidants. An additional suitable pharmaceutically acceptable excipient may be a chelating agent. Tablet formulations may be uncoated or coated to modify their disintegration and subsequent absorption of the active ingredient in the gastrointestinal tract, or to improve their stability and / or appearance, in any case using conventional coating agents and procedures well known. from the prior art.

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

Aqueous suspensions generally contain the active ingredient in finely divided form together with one or more suspending, dispersing or wetting agents. Aqueous suspensions may also contain one or more preservatives, antioxidants, colors, flavors and / or sweeteners.

Oily suspensions can be formulated by suspending the active ingredient in vegetable oil or mineral oil. Oily suspensions may also contain a thickening agent. Sweeteners such as those set forth above and flavoring agents can be added to provide an oral preparation with an attractive taste. Such compositions can be preserved by the addition of an antioxidant.

Dispersible powders and granules suitable for preparing an aqueous suspension by adding water generally contain the active ingredient together with a dispersing or wetting agent, a suspending agent and one or more preservatives. Additional auxiliaries such as sweeteners, flavors and colors may also be present.

Pharmaceutical compositions in accordance with the present disclosure can also be presented in the form of oil-in-water emulsions. The oily phase can be vegetable oil or mineral oil, or a mixture of any of these. Emulsions can also contain sweeteners, flavors and preservatives.

Syrups and elixirs can be formulated with sweeteners and can also contain an emollient, preservative, flavoring and / or coloring agent.

The pharmaceutical compositions can also be presented in the form of a sterile injectable aqueous or oily suspension, which can be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents as mentioned above. The sterile injectable may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent system.

Compositions to be administered by inhalation may be in the form of a conventional pressurized aerosol for dispensing the active ingredient in the form of an aerosol containing fine solid particles or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons can be used, and the aerosol device is conveniently arranged to dispense a metered amount of active ingredient. Dry powder inhalers may also be suitable.

For additional compilation information, the reader is referred to Chapter 25.2 of Volume 5 of the Comprehensive Medicinal Chemistry (Corwin Hansch; editorial board chairman), Pergamon Press 1990.

The amount of active ingredient, which is combined with one or more excipients to obtain a unit dosage form, will inevitably vary in

- 32 037533 depending on the recipient being treated and the particular route of administration. For example, oral administration to humans will generally require administration of, for example, 1 mg to 2 g of active agent (more suitably 100 mg to 2 g, such as 250 mg to 1.8 g, such as , from 500 mg to 1.8 g, in particular from 500 mg to 1.5 g, for convenience purposes from 500 mg to 1 g), formulated as a mixture with an appropriate and appropriate amount of excipients, which may vary from about 3 up to about 98% by weight of the total composition. It should be understood that in the event that a large dose is required, several dosage forms may be required, for example two or more tablets or capsules, with the dose of the active ingredient dispersed therebetween for convenience sake. Typically, unit dosage forms will contain from about 10 mg to 0.5 g of a compound as described herein, although a unit dosage form may contain up to 1 g. For convenience, a unit solid dosage form may contain from 1 mg to 300 mg of active ingredient. ...

The dosage size of the compounds as described herein for therapeutic or prophylactic purposes will naturally vary according to the nature and severity of the disease condition, the age and sex of the animal or patient, and the route of administration in accordance with well-known medical principles.

When the compounds are used according to the present disclosure for therapeutic or prophylactic purposes, they are usually administered so that the daily dose is in the range of, for example, 1 to 100 mg / kg body weight, optionally in divided doses. Typically, when using the parenteral route, lower doses will be administered. Thus, for example, for intravenous administration, a dose in the range of, for example, 1 to 25 mg / kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range of, for example, 1 to 25 mg / kg body weight will be used. However, oral administration is preferred, in particular in the form of tablets.

In one aspect of the present description, the compounds in accordance with the present description or their pharmaceutically acceptable salts are administered in the form of tablets containing from 10 to 100 mg of the compound in accordance with the present description (or a pharmaceutically acceptable salt thereof), where to achieve the required dose, depending on if necessary, one or more tablets are administered.

As indicated above, ERa signaling is known to induce tumor formation through one or more of the effects of mediating proliferation of cancer and other cells, mediating angiogenesis events, and mediating cancer cell motility, migration and invasiveness. The present inventors have found that the compounds according to the present disclosure have potent antitumor activity, which is believed to be achieved through antagonizing and suppressing ERa, which is involved in the signaling steps that mediate tumor cell proliferation and survival, as well as invasiveness. and the ability to migrate in metastatic tumor cells.

Accordingly, the compounds according to the present disclosure may be of value as antitumor agents, in particular as selective inhibitors of proliferation, survival, motility, dissemination and invasiveness of mammalian cancer cells, leading to suppression of tumor growth and survival and suppression of the growth of metastatic tumors. In particular, compounds according to the present disclosure may be of value as antiproliferative and antiinvasive agents in containing and / or treating a disease characterized by the presence of a solid tumor. In particular, the compounds according to the present disclosure may be useful in the prevention or treatment of those tumors that are sensitive to the suppression of ERa function, which are involved in the signaling steps involved in the proliferation and survival of tumor cells, as well as the ability to migrate and invasiveness. in metastatic tumor cells. In addition, compounds according to the present disclosure may be useful in the prevention or treatment of those tumors for which it is mediated solely or partially by ERa antagonism and suppression, i. E. the compounds can be used to obtain an ERa inhibitory effect in a warm-blooded animal in need of such treatment.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use as a medicament in a warm-blooded animal, such as a human.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in producing an antiproliferative effect in a warm blooded animal, such as a human.

In accordance with a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in a warm-blooded animal, such as a human, as an anti-invasive agent in containing and / or treating a disease characterized by the presence of solid tumor.

- 33 037533

In accordance with a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, to obtain an antiproliferative effect in a warm blooded animal, such as a human.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in obtaining an antiproliferative effect in a warm blooded animal, such as a human.

In accordance with a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in a warm-blooded animal, such as a human, as an anti-invasive agent in containment and / or treating a disease characterized by the presence of a solid tumor.

In accordance with a further aspect of the present disclosure, there is provided a method of obtaining an antiproliferative effect in a warm blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

In accordance with a further aspect of the present disclosure, there is provided a method of obtaining an anti-invasive effect by containing and / or treating a disease characterized by the presence of a solid tumor in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID ) or a pharmaceutically acceptable salt thereof, as defined above in this document.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the prevention or treatment of cancer in a warm-blooded animal, such as a human.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the prevention or treatment of cancer in a warm-blooded animal, such as a human.

According to a further aspect of the present disclosure, there is provided a method for preventing or treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the prevention or treatment of a disease characterized by the presence of a solid tumor in a warm-blooded animal, such as a human.

In accordance with a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the prevention or treatment of a disease characterized by the presence of a solid tumor in a warm-blooded animal such as a human ...

According to a further aspect of the present disclosure, there is provided a method for preventing or treating a disease characterized by the presence of a solid tumor in a warm-blooded animal, such as a human, in need of such treatment, comprising administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, defined above in this document.

In accordance with a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined herein, for use in the prevention or treatment of tumors that are sensitive to suppression of ERa function, which are involved in signaling steps leading to proliferation, survival, invasiveness and the ability to migrate tumor cells.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the prevention or treatment of tumors that are sensitive to suppression of ERa function that are involved in the stages of signal transduction that determine the proliferation, survival, invasiveness and the ability to migrate tumor cells.

In accordance with a further aspect of the present disclosure, there is provided a method for preventing or treating tumors that are sensitive to suppression of ERa function that are involved in the signaling steps involved in proliferation, survival, invasiveness and migration of tumor cells, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in providing an ERa inhibitory effect.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in providing an ERa inhibitory effect.

In accordance with a further aspect of the present disclosure, there is also provided a method of providing an ERa inhibitory effect, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in providing a selective inhibitory effect on ERa.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in providing a selective inhibitory effect on ERa.

In accordance with a further aspect of the present disclosure, there is also provided a method of providing a selective inhibitory effect on ERa, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

This document describes compounds that can bind to the ligand binding domain of ERa and are selective estrogen receptor disruptors. Biochemical and cellular assays have shown that compounds according to the present disclosure are active in binding to estrogen receptors and reduce cellular levels of ERa, and therefore may be useful in the treatment of estrogen-sensitive diseases or conditions (including diseases characterized by the emergence of resistance to endocrine therapy drugs), i.e. for use in the treatment of breast and gynecological cancers (including endometrium, ovarian and cervical cancers) and cancers expressing ERa proteins with mutations that may be de novo mutations or those resulting from treatment with more an early endocrine therapy drug such as an aromatase inhibitor.

In accordance with a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the treatment of breast cancer or gynecological cancers.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the treatment of breast, endometrial, ovarian, or cervical cancer.

In accordance with a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the treatment of breast cancer.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the treatment of breast cancer where the cancer cells have developed resistance to one or more other endocrine therapy drugs.

In accordance with a further aspect of the present disclosure, there is provided a method of treating breast or gynecological cancers, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

In accordance with a further aspect of the present disclosure, there is provided a method of treating breast, endometrial, ovarian, or cervical cancer, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

In accordance with a further aspect of the present disclosure, there is provided a method of treating breast cancer, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

In accordance with a further aspect of the present disclosure, there is provided a method of treating breast cancer, wherein the cancer cells have developed resistance to one or more of the other

35 037533 endocrine therapy, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the treatment of breast cancer or gynecological cancers.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the treatment of breast, endometrial, ovarian, or cervical cancer.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the treatment of breast cancer.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the treatment of breast cancer where the cancer cells have developed resistance to one or more other drugs endocrine therapy.

In one feature of the present disclosure, the cancer to be treated is breast cancer. In an additional aspect of this trait, breast cancer is estrogen receptor positive (ER positive). In one embodiment of this aspect, a compound of formula (ID) is administered together with another anti-cancer agent, such as an anti-hormonal agent as defined herein.

According to a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, for use in the treatment of ER-positive breast cancer.

According to a further aspect of the present disclosure, there is provided a method of treating ER-positive breast cancer, which comprises administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, as defined hereinabove, in the manufacture of a medicament for use in the treatment of ER-positive breast cancer.

As indicated above in this document, the in vivo effects of the compound of formula (ID) may in part be due to one or more metabolites that are formed in the human or animal body after administration of the compound of formula (ID).

The present disclosure therefore also contemplates a method for suppressing ER-α function in a patient, comprising administering to the patient an amount of a compound of formula (ID) or a pharmaceutically acceptable salt thereof effective to suppress ER-α function in the patient.

The present disclosure therefore also contemplates a method for suppressing ER-α function in a patient, comprising administering to the patient an amount of a compound of formula (ID) or a pharmaceutically acceptable salt thereof effective to suppress ER-α function in the patient.

The anticancer treatment as defined herein can be used as monotherapy or can include, in addition to using compounds as described herein, conventional surgery or radiation therapy or chemotherapy. Such chemotherapy may involve the use of one or more of the following categories of antineoplastic agents:

(i) other antiproliferative / antineoplastic drugs and combinations thereof used in medical oncology, such as alkylating agents (for example, cisplatin, oxaliplatin, carboplatin, cyclophosphamide, chlormethine, melphalan, chlorambucil, busulfan, temozolomide and nitrosoureas); antimetabolites (eg gemcitabine and antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside and hydroxyurea); antineoplastic antibiotics (eg, anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin); antimitotic agents (eg vinca alkaloids such as vincristine, vinblastine, vindesine and vinorelbine, as well as taxoids such as taxol and taxotere and polo kinase inhibitors) and topoisomerase inhibitors (eg epipodophyllotoxins such as etoposide and tenena amsacrine, topotecan, and camptothecin);

(ii) antihormonal agents such as antiestrogens (eg tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, and iodoxifene), progestogens (eg megestrol acetate), in-

- 36 037533 aromatase inhibitors (for example, such as anastrozole, letrozole, vorazole and exemestane);

(iii) inhibitors of growth factor function and their downstream signaling pathways: Ab modulators of any growth factor or growth factor receptor targets reviewed by Stern et al are included. Critical Reviews in Oncology / Haematology, 2005, 54, p. 11-29); also includes small molecule inhibitors of such targets, for example, kinase inhibitors - examples include trastuzumab, which is an anti-erbB2 antibody [Herceptin ™], panitumumab, which is an antibody to EGFR, cetuximab, which is an antibody to EGFR [erbitux, C225 inhibitors], and including inhibitors of the erbB family of receptors, such as inhibitors of tyrosine kinases belonging to the epidermal growth factor receptor (EGFR / erbBl) family, such as gefitinib or erlotinib, inhibitors of erB2 tyrosine kinase such as lapatinib, and mixed inhibitors of erb1 / 2, ; similar strategies are available for other classes of growth factors and their receptors, for example inhibitors of the hepatocyte growth factor family or their receptors, including c-met and ron; inhibitors of insulin and the family of insulin growth factors or their receptors (IGFR, IR), inhibitors of the family of growth factors of platelets or their receptors (PDGFR) and inhibitors of signaling mediated by other receptor tyrosine kinases such as c-kit, AnLK and CSF-1R; also included modulators targeting signaling proteins in the PI3 kinase signaling pathway, e.g. inhibitors of PI3 kinase isoforms such as PI3K-a / e / y and ser / thr kinases such as AKT, mTOR (such as AZD2014) , PDK, SGK, PI4K, or PIP5K; also included are inhibitors of serine-threonine kinases not listed above, for example, raf inhibitors such as vemurafenib, MEK inhibitors such as selumetinib (AZD6244), Abl inhibitors such as imatinib or nilotinib, Btk inhibitors such as ibrutinib, such as phytarafenib inhibitors , aurorakinase inhibitors (eg AZD1152), inhibitors of other ser / thr kinases such as JAK species, STAT species and IRAK4, and cyclin dependent kinase inhibitors, such as CDK1, CDK7, CDK9 and CDK4 / 6 inhibitors such as palbociclib;

iv) modulators of DNA damage signaling pathways, eg PARP inhibitors (eg olaparib), ATR inhibitors or ATM inhibitors;

v) modulators of signaling pathways of apoptosis and cell death, such as modulators of the Bcl family (eg, ABT-263 / navitoclax, ABT-199);

(vi) anti-angiogenic agents, such as agents that inhibit the effects of vascular endothelial growth factor [for example, bevacizumab, which is an antibody to vascular endothelial cell growth factor (Avastin ™), and, for example, a VEGF receptor tyrosine kinase inhibitor such as sorafenib, axitinib, pazopanib, sunitinib and vandetanib, as well as compounds acting through other mechanisms (eg, linomide, inhibitors of integrin ave3 function, and angiostatin)];

(vii) vascular damaging agents such as combretastatin A4;

(viii) anti-invasive agents, for example, c-Src family kinase inhibitors such as dasatinib (J. Med. Chem., 2004, 47, 6658-6661) and bosutinib (SKI-606), as well as metalloproteinase inhibitors such as marimastat, inhibitors of urokinase-type plasminogen activator receptor function or anti-heparanase antibodies;

(ix) immunotherapeutic approaches, including, for example, ex vivo and in vivo approaches to increase the immunogenicity of tumor cells in a patient, such as transfection with cytokines such as interleukin-2, interleukin-4, or granulocyte macrophage colony stimulating factor, approaches to reduce T -cellular tolerance, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumor cell lines, and approaches using anti-idiotypic antibodies. Specific examples include monoclonal antibodies that target PD-1 (eg, BMS-936558) or CTLA4 (eg, ipilimumab and tremelimumab);

(x) types of antisense or RNAi-based therapies, for example, directed to the listed targets;

(xi) gene therapy approaches, including, for example, approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene-mediated enzyme prodrug therapy) approaches such as cytosine deaminase, or thymidine kinase approaches , and approaches to increasing patient tolerance to chemotherapy or radiation therapy, such as multidrug resistance gene therapy.

Accordingly, in one embodiment, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and an additional anticancer agent for the joint treatment of cancer.

In accordance with this aspect of the present disclosure, there is provided a combination suitable for use in the treatment of cancer comprising a compound of formula (ID) or a pharmaceutically acceptable salt thereof and another anticancer agent, in particular any of the antineoplastic agents listed in items (i) to (xi) above. In particular, the use of the antineoplastic agents listed in items (i) to (xi) above is the standard of care for the particular type of cancer to be treated; a person skilled in the art will understand the meaning of the term standard of care.

Therefore, in a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with another antineoplastic agent, in particular an antineoplastic agent selected from those recited in items (i) (xi) above herein.

In a further aspect of the present disclosure, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided in combination with another antineoplastic agent, in particular an antineoplastic agent selected from those listed in (i) above.

In a further aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and any of the antineoplastic agents listed in paragraph (i) above.

In a further aspect of the present disclosure, a combination suitable for use in the treatment of cancer is provided, comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a taxoid such as, for example, Taxol or Taxotere, preferably Taxotere.

In a further aspect of the present disclosure, a compound of formula (ID), or a pharmaceutically acceptable salt thereof, is provided in combination with another antineoplastic agent, in particular an antineoplastic agent selected from those listed in item (ii) above herein.

In a further aspect of the present disclosure, a combination suitable for use in the treatment of cancer is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and any of the anti-hormonal agents listed in paragraph (ii) above, for example, any of the antiestrogens listed in paragraph ( ii) above, or, for example, from the aromatase inhibitors listed in item (ii) above.

In a further aspect of the present disclosure, a combination suitable for use in the treatment of cancer is provided, comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and an mTOR inhibitor such as AZD2014.

In a further aspect of the present disclosure, a combination suitable for use in the treatment of cancer is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a PI3Kα inhibitor, such as compound 1- (4- (5- (5-amino-6- ( 5-tert-butyl-1,3,4-oxadiazol-2yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4-triazol-3-yl) piperidin-1-yl) -3- hydroxypropan-1-one; or a pharmaceutically acceptable salt thereof.

In a further aspect of the present disclosure, a combination suitable for use in the treatment of cancer is provided, comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a CDK4 / 6 inhibitor such as palbociclib.

In one aspect, the above combination of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, with an antineoplastic agent listed in (ii) above, or an mTOR inhibitor (such as AZD2014), or a PI3K-a inhibitor (such as compound 1- (4- (5- (5-amino-6- (5-tert-butyl-1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4-triazole-3- yl) piperidin-1-yl) -3hydroxypropan-1-one), or a CDK4 / 6 inhibitor (such as palbociclib) is suitable for use in the treatment of breast or gynecological cancers such as breast, endometrial, ovarian or cervical cancer, in particular breast cancer such as ER positive breast cancer.

Whenever the term combination is used herein, it should be understood to refer to simultaneous, separate or sequential administration. In one aspect of the present disclosure, the combination refers to simultaneous administration. In another aspect of the present disclosure, the combination refers to separate administration. In a further aspect of the present disclosure, the combination refers to sequential administration. If the administration is sequential or separate, then the delay in the administration of the second component should not be such that the beneficial effect of the combination is lost. If a combination of two or more components is administered separately or sequentially, it will be appreciated that the dosage regimen for each component may be different from and independent of the dosage regimens of the other components. For convenience, compounds as described herein are dosed once a day.

In accordance with a further aspect of the present disclosure, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent selected from those listed in items (i) to (xi) above herein, in combination with a pharmaceutically acceptable excipient.

In accordance with a further aspect of the present disclosure, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with any of the anti-hormonal agents listed in paragraph (ii) above, for example any of the antiestrogens listed in item (ii) above, or, for example, of the aromatase inhibitors listed in item (ii) above, in combination with a pharmaceutically acceptable excipient.

In a further aspect of the present disclosure, a pharmaceutical composition is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and an mTOR inhibitor such as AZD2014, in combination with a pharmaceutically acceptable excipient.

In an additional aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a PI3Kα inhibitor, such as compound 1- (4- (5- (5-amino-6- (5-tert-butyl- 1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H1,2,4-triazol-3-yl) piperidin-1-yl) -3-hydroxypropan-1-one, in combination with a pharmaceutically acceptable excipient.

In an additional aspect of the present disclosure, a pharmaceutical composition is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a CDK4 / 6 inhibitor (such as palbociclib) in combination with a pharmaceutically acceptable excipient.

In accordance with a further aspect of the present disclosure, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent selected from those listed in items (i) to (xi) above herein, in combination with a pharmaceutically acceptable excipient for use in the treatment of cancer.

According to a further aspect of the present disclosure, a pharmaceutical composition is provided that comprises a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with any of the anti-hormonal agents listed in paragraph (ii) above, for example, any of the antiestrogens listed in paragraph (ii) above, or, for example, of the aromatase inhibitors listed in item (ii) above, in combination with a pharmaceutically acceptable excipient for use in the treatment of cancer.

In a further aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and an mTOR inhibitor, such as AZD2014, in combination with a pharmaceutically acceptable excipient for use in the treatment of cancer.

In an additional aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a PI3Kα inhibitor, such as compound 1- (4- (5- (5-amino-6- (5-tert-butyl- 1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H1,2,4-triazol-3-yl) piperidin-1-yl) -3-hydroxypropan-1-one, in combination with a pharmaceutically acceptable excipient for use in the treatment of cancer.

In a further aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, and a CDK4 / 6 inhibitor (such as palbociclib) in combination with a pharmaceutically acceptable excipient for use in the treatment of cancer.

In one aspect, the aforementioned pharmaceutical compositions based on a compound of formula (ID), or a pharmaceutically acceptable salt thereof, with an antitumor agent listed in item (ii) above, or an mTOR inhibitor (such as AZD2014), or a PI3K-a inhibitor (such as compound 1- (4- (5- (5-amino-6- (5-tert-butyl-1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4triazole -3-yl) piperidin-1-yl) -3-hydroxypropan-1-one), or a CDK4 / 6 inhibitor (such as palbociclib) are suitable for use in the treatment of breast cancer or gynecological cancers such as breast cancer , endometrial, ovarian, or cervical cancer, in particular breast cancer such as ER-positive breast cancer.

In accordance with another feature of the present description, there is provided the use of a compound of formula (ID) or a pharmaceutically acceptable salt thereof in combination with an antineoplastic agent selected from those listed in items (i) to (xi) above in this document, in the manufacture of a medicament intended for use in the treatment of cancer in a warm-blooded animal such as humans.

According to a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with any of the anti-hormonal agents listed in (ii) above, for example, any of the antiestrogens listed in (ii) above, or, for example, from the aromatase inhibitors listed in item (ii) above, in the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal such as a human.

In a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an mTOR inhibitor such as AZD2014 in the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal, such as a human.

- 39 037533

In a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a PI3Kα inhibitor such as compound 1- (4- (5- (5-amino-6- (5-tert-butyl-1 , 3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-III-1,2,4-triazol-3yl) piperidin-1-yl) -3-hydroxypropan-1-one, c the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal, such as a human.

In a further aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a CDK4 / 6 inhibitor (such as palbociclib) in the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal, such as a human.

In one aspect, the aforementioned uses of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent listed in (ii) above, or an mTOR inhibitor (such as AZD2014), or a PI3K-a inhibitor (such as Compound 1- (4- (5- (5-amino-6- (5-tert-butyl-1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4 -triazol-3yl) piperidin-1-yl) -3-hydroxypropan-1-one), or a CDK4 / 6 inhibitor (such as palbociclib) are suitable for use in the manufacture of a medicament for the treatment of breast or gynecological cancers such as breast, endometrial, ovarian or cervical cancer, in particular breast cancer such as ER positive breast cancer.

Therefore, an additional feature of the present disclosure provides a method for treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent selected from items (i) - (xi) above in this document.

In accordance with a further aspect of the present disclosure, there is provided a method of treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with any of the anti-hormonal agents listed in item (ii) above, for example, any of the antiestrogens listed in item (ii) above, or, for example, from the aromatase inhibitors listed in item (ii) above.

In a further aspect of the present disclosure, there is provided a method of treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an mTOR inhibitor such as AZD2014.

In a further aspect of the present disclosure, there is provided a method of treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a PI3Kα inhibitor such as Compound 1 - (4- (5- (5-amino-6- (5-tert-butyl1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4- triazol-3-yl) niperidin-1-yl) -3-hydroxynropan-1-one.

In a further aspect of the present disclosure, there is provided a method of treating cancer in a warm-blooded animal, such as a human, in need of such treatment, which comprises administering to said animal an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with a CDK4 / 6 inhibitor (such as palbociclib ).

In one aspect, the above combinations, pharmaceutical compositions, uses and methods for treating cancer are methods for treating breast or gynecological cancers such as breast, endometrial, ovarian or cervical cancers, in particular breast cancers such as ER- positive breast cancer.

In accordance with a further aspect of the present disclosure, a kit is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent selected from those recited in items (i) to (xi) above herein.

According to a further aspect of the present disclosure, a kit is provided comprising a compound of formula (ID), or a pharmaceutically acceptable salt thereof, in combination with an antineoplastic agent selected from those listed in (i) or (ii) above herein.

In accordance with a further aspect of the present disclosure, a kit is provided comprising:

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

b) an antineoplastic agent selected from those listed in items (i) - (xi) above in this document, in the form of a second unit dosage form; and

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

- 40 037533

In accordance with a further aspect of the present disclosure, a kit is provided comprising:

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

b) an antineoplastic agent selected from those listed in items (i) - (ii) above in this document, in the form of a second unit dosage form;

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

In accordance with a further aspect of the present disclosure, a kit is provided comprising:

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

b) an antineoplastic agent selected from the antineoplastic agents listed in item (ii) above, an mTOR inhibitor (such as AZD2014), a PI3Kα inhibitor such as compound 1- (4- (5- (5amino-6- (5-tert -butyl-1,3,4-oxadiazol-2-yl) pyrazin-2-yl) -1-ethyl-1H-1,2,4-triazol-3-yl) piperidin-1yl) -3-hydroxypropane-1 -one and a CDK4 / 6 inhibitor such as palbociclib in a second unit dosage form;

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

The combination therapy described above can be carried out in addition to standard treatment, usually carried out in accordance with his usual schedule of administration.

Although the compounds of formula (ID) are generally of value as therapeutic agents for use in warm-blooded animals (including humans), they are also useful in all cases where it is necessary to suppress ERa function. Thus, they are applicable as pharmacological standards for use in the development of new biological tests and in the search for new pharmacological agents.

Personalized medical care.

Another aspect of the present disclosure is based on the identification of the relationship between the status of the gene encoding ERa and the potential response to treatment with a compound of formula (ID). In particular, ERa gene status may indicate that the patient is less likely to respond to existing hormone therapy (such as aromatase inhibitors), at least in part, because some ERa mutations are thought to arise as resistance mechanisms to existing types of treatment. SERD, in particular SERD, which can be administered orally in potentially large doses without undue inconvenience, can therefore be advantageously used to treat patients with ERa mutations who may be resistant to other therapies. Thus, it provides opportunities, methods and tools for selecting patients for treatment with a compound of formula (ID), in particular cancer patients. This description relates to tools and methods for selecting patients (including personalized medicine). The selection is based on whether the tumor cells to be treated contain the wild-type or mutant ERa gene. The ERa gene status can therefore be used as a biomarker to indicate that the choice of SERD treatment may be appropriate. For the avoidance of doubt, it is believed that the compounds of formula (ID) described herein exhibit similar activity against the wild-type ERa gene and the mutant ERa gene, at least the ERa gene with mutations identified at the filing date of this application.

There is a clear need for biomarkers that will expand the category or select patients whose tumors will respond to SERD treatment, such as a compound of formula (ID). Patient selection biomarkers that identify patients most likely to respond to one drug over others are ideal for cancer treatment because they reduce the incidence of unnecessary treatment for patients with non-responsive tumors and the potential side effects of such drugs.

A biomarker can be described as a characteristic parameter that can be objectively measured and assessed as an indicator of normal biological processes, pathogenic processes, or pharmacological responses to therapeutic intervention. A biomarker is any identifiable and measurable indicator associated with a specific condition or disease for which there is a correlation between the presence or level of a biomarker and some aspect of the condition or disease (including the presence, level or change in level, type, stage of the condition or disease, predisposition to it or the ability to respond to a drug used to treat a condition or disease). Correlation can be qualitative, quantitative, or both qualitative and quantitative. Typically, a biomarker is a compound, a fragment of a compound, or a group of compounds. Such compounds can be any compound found or produced in the body, including proteins (and peptides), nucleic acids, and other compounds.

Biomarkers can have predictive potential and, as such, can be used for

- 41 037533 predicting or determining the presence, level, type or stage of specific conditions or diseases (including the presence or level of specific microorganisms or toxins), predisposition (including genetic predisposition) to specific conditions or diseases, or response to specific treatments ( including types of drug treatment). It is believed that biomarkers will play an increasingly important role in future drug discovery and development by improving the effectiveness of research and development programs. Biomarkers can be used as diagnostic tools, indicators of disease progression, indicators of treatment, and predictors of clinical outcome. For example, various biomarker research projects are attempting to identify markers for certain cancers and certain cardiovascular and immunological diseases. The development of new validated biomarkers is believed to result in both significant reductions in health care and drug development costs and significant improvements in the treatment of a wide range of diseases and conditions.

A biomarker may be required for optimal planning of clinical trials and to obtain the most complete information from these trials. The marker can be measurable in mimic and tumor tissues. Ideally, these markers will also correlate with efficacy and thus ultimately can be used for patient selection.

Thus, the technical problem underlying this aspect of the present disclosure is to define a method for stratifying patients by treatment with a compound of formula (ID). This technical problem is solved by providing for the embodiments described in this document in the claims and / or in the description.

Tumors that contain wild-type ERa are believed to be susceptible to treatment with a compound of formula (ID), for example, as a first-line treatment. Tumors may also respond to treatment with a compound of formula (ID) as second-line, third-line or follow-up therapy, and this can be used in particular if the tumors contain a mutant ERa and can thus be resistant to existing therapies, such like AI. Treatment of resistant tumors may require a higher dose of a compound of formula (ID) than tumors expressing the wild-type receptor.

Provided herein is a method for determining the sensitivity of cells to a compound of formula (ID). This method includes determining the status of the ERa gene in these cells. A cell is defined as responsive to a compound of formula (ID) if it inhibits an increase in the number of cells in a cell growth assay (by inhibiting cell proliferation and / or by increasing the rate of cell death). The methods described herein are useful for predicting which cells are more likely to respond to a compound of formula (ID) by inhibiting growth.

The tumor characterizing sample can be an actual isolated tumor sample, or it can be a sample that has been further processed, for example, a sample of PCR amplified nucleic acid from a tumor sample.

Definitions.

In this section on Personalized Care, the following is accepted.

An allele refers to a specific form of a genetic locus that differs from other forms in its specific nucleotide or amino acid sequence.

Amplification reactions are nucleic acid reactions that result in the specific amplification of target nucleic acids among non-target nucleic acids. Polymerase chain reaction (PCR) is a well-known amplification reaction.

As used herein, cancer refers to neoplastic growth resulting from the transformation of cells into a neoplastic phenotype. Genetic mutation is often involved in this cell transformation.

A gene is a segment of DNA that contains all the information necessary for the regulated biosynthesis of an RNA product, including a promoter, exons, introns, and other expression control sequence elements that can be located within the 5'- or 3'-flanking sites (but not in the transcribed parts of the gene).

Gene status refers to whether a gene is wild-type or not (i.e., mutant).

A label refers to a composition capable of producing a detectable signal indicative of the presence of a target polynucleotide in a sample to be analyzed. Suitable labels include radioactive isotopes, nucleotide chromophores, enzymes, substrates, fluorescent molecules, chemiluminescent moieties, magnetic particles, bioluminescent moieties, and the like. Thus, a label is any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical methods.

A nonsynonymous change refers to a change (variation) in the coding sequence of a gene or overlapping region that results in a different (altered) polypeptide sequence. These changes may or may not affect protein function and include missense variants (where one amino acid is substituted for another), nonsense variants (where the result is polypeptide truncation due to premature stop codon formation), and insertion-deletion options.

Synonymous change refers to a change (variation) in the coding sequence of a gene that does not affect the sequence of the encoded polypeptide. These changes can indirectly affect protein function (for example, by altering gene expression), but in the absence of evidence to the contrary, they are generally considered harmless.

Nucleic acid refers to single-stranded or double-stranded DNA and RNA molecules, including naturally occurring nucleic acids found in nature and / or modified artificial nucleic acids having modified backbones or nucleic bases, as is known in the art.

A primer refers to a single-stranded DNA oligonucleotide sequence capable of acting as an initiation point for synthesizing a primer extension product that is complementary to the nucleic acid strand to be copied. The length and sequence of the primer must be such that it can prime the synthesis of extension products. A typical primer is at least about 7 nucleotides in length and is substantially complementary to the target sequence, but slightly longer primers are preferred. Typically, primers are about 15-26 nucleotides in length, but longer or shorter primers can also be used.

A polymorphic site is a position at a locus at which at least two alternative sequences are found in a population.

Polymorphism refers to a sequence change observed in an individual at a polymorphic site. The types of polymorphism include nucleotide substitutions, insertions, deletions, and microsatellites and may, but need not be, cause detectable differences in gene expression or protein function. In the absence of evidence of an effect on protein expression or function, common polymorphism types, including nonsynonymous variants, are generally considered to be included in wild-type gene sequencing.

A catalog of human polymorphism types and related annotations, including validation, frequency of observation, and disease association, is maintained by the NCBI (dbSNP: http://www.ncbi.nlm.nih.gov/projects/SNP/). It should be noted that the term polymorphism when used in relation to gene sequences should not be confused with the term polymorphism when used in relation to the solid-state form of a compound, i.e. crystalline or amorphous nature of the compound. A person skilled in the art will understand the intended meaning from its context.

Probe refers to sequence-specific single-stranded oligonucleotides that have a sequence exactly complementary to the target allele sequence to be detected.

Response is determined by measurements made in accordance with the criteria for evaluating the response of solid tumors (RECIST), which classifies patients into two main groups: those showing a partial response or stabilization of the disease, and those showing signs of disease progression.

Stringent hybridization conditions refer to overnight incubation at 42 ° C in a solution containing 50% formamide, 5x SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10 % dextran sulfate and 20 pg / ml denatured hydrodynamically shear dehydrated DNA from salmon milk, followed by washing the filters in 0.1x SSC at approximately 65 ° C.

Survival encompasses overall survival and progression-free survival in a patient.

Overall survival (OS) is defined as the time from initiation of drug administration to death for any reason. Progression-free survival (PFS) is defined as the time from initiation of drug administration to the first manifestation of disease progression or death from any cause.

In accordance with one aspect of the present disclosure, there is provided a method of selecting patients for treatment with a compound of formula (ID), the method comprising obtaining from a patient a sample that contains tumor cells;

determining whether the ERa gene in the tumor cell-containing sample obtained from the patient is wild-type or mutant; and selecting a patient for treatment with a compound of formula (ID) based on this.

The method may or may not include the step of actually taking a sample from the patient. Thus, in accordance with one aspect of the present disclosure, there is provided a method for selecting a patient for treatment with a compound of formula (ID), the method comprising determining whether the ERa gene in a sample containing tumor cells previously isolated from a patient is a gene wild type or mutant; and selecting a patient for treatment with a compound of formula (ID) based on this.

In one embodiment, the patient is selected for treatment with a compound of formula (ID) if the tumor cell DNA contains the mutant ERa gene. In other embodiments, a patient whose tumor cell DNA contains a wild-type ERa gene is selected for treatment with a compound of formula (ID).

For the purposes of this disclosure, wild-type gene status is intended to indicate normal or appropriate gene expression and normal function of the encoded protein. In contrast, mutant status is intended to indicate the expression of a protein with altered function in accordance with the known roles of mutant ERa genes in cancer (as described herein). Any number of genetic or epigenetic changes, including but not limited to mutation, amplification, deletion, genomic rearrangement, or changes in the methylation profile, can result in mutant status. However, if such changes, in spite of everything, lead to the corresponding expression of a normal protein or a functionally equivalent variant, then the status of the gene is considered to be wild-type status. Examples of variants that usually do not cause the occurrence of a functional mutant status of a gene include synonymous coding variants and types of common polymorphisms (synonymous or nonsynonymous). As discussed below, the status of a gene can be assessed by functional analysis, or it can be inferred from the nature of the identified deviations from the reference sequence.

In certain embodiments, wild-type or mutant ERa gene status is determined by the presence or absence of nonsynonymous nucleic acid changes in the genes. The observed nonsynonymous changes, corresponding to known types of common polymorphism without annotated functional effects, do not contribute to the emergence of the mutant status of the gene.

Other changes in the ERa gene that indicate mutant status include splice site changes that reduce recognition of the intron-exon boundary during pre-mRNA-to-mRNA conversion. This can lead to exon skipping or normal incorporation of the intron sequence into the spliced mRNA (intron retention or the use of hidden splicing boundaries). In turn, this can lead to the production of an aberrant protein with insertions and / or deletions relative to the normal protein. Thus, in other embodiments, a gene is mutated if a variant is present that alters the sequence of splice site recognition at the intron-exon interface.

In the case of ESR1, reference sequences are available for the gene (GenBank accession: NG_008493), mRNA (GenBank accession number: NM_000125) and protein (GenBank accession number: NP_000116 or Swiss-Prot accession: P03372). One skilled in the art will be able to determine the status of the ESR1 gene, i. E. whether a particular ESR1 gene is a wild-type gene or a mutant gene, based on a comparison of the DNA or protein sequence with the wild-type one.

It will be apparent that the gene and mRNA sequences disclosed for the ERa gene are exemplary sequences. In normal individuals, there are two copies of each gene, maternal and paternal copies, which are likely to have some sequence differences; moreover, there are numerous allelic variants of a given gene sequence in the population. Other sequences that are considered wild-type include those that have one or more synonymous changes in the nucleic acid sequence (and these changes do not change the sequence of the encoded protein), nonsynonymous types of common polymorphism (for example, types of polymorphism in germ-line cells) that change protein sequence, but not affecting protein function, and changes in intron sequences not affecting the splicing site.

To determine the status of the ERa gene, there are many techniques available to the person skilled in the art. Gene status can be determined by determining the nucleic acid sequence. This can be done by direct sequencing of the full-length gene or by analyzing specific sites in the gene, for example, sites that are frequently mutated.

Samples.

The patient sample to be tested for gene status can be any sample containing tumor tissue or tumor cells obtained or obtained from an individual. For convenience, the test sample is a blood sample, oral swab, biopsy, or other biological fluid or tissue obtained from an individual. Specific examples include circulating tumor cells, circulating DNA in plasma or serum, cells isolated from ascitic fluid from patients suffering from ovarian cancer, pulmonary sputum from patients with lung tumors, fine-needle punctate obtained from a patient suffering from breast cancer, urine , peripheral blood, scraping of cells, hair follicle, a skin sample obtained by pinch biopsy, or a sample of buccal epithelium.

It will be understood that the test sample may equally be a nucleic acid sequence corresponding to the sequence in the test sample, which means that all or part of a region in a nucleic acid sample can be first amplified prior to analysis using any convenient technique, for example, polymerase chain reaction (PCR). The nucleic acid can be genomic DNA or fractionated or total cellular RNA. In specific embodiments, the RNA is total cellular RNA and is used directly as a template for labeling the first strand to DNA using random primers or poly-A containing primers. The nucleic acid or protein in the test sample can be extracted from the sample according to standard techniques (see Green & Sambrook, Eds., Molecular Cloning: A Laboratory Manual, (2012, 4th ed., Volume 1-3, ISBN 9781936113422) , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY).

The diagnostic methods of the present disclosure can be performed using a sample previously obtained from an individual or patient. Such samples can be preserved by freezing or fixing and embedded in formalin / paraffin or other media. Alternatively, a fresh sample containing tumor cells can be obtained and used.

The methods in accordance with the present description can be applied using cells from any tumor. Suitable tumors for treatment with a compound of formula (ID) have been described above herein.

Methods for detecting nucleic acids.

As used herein, the detection of mutant ERa nucleic acids can be used to guide drug treatment. Since mutations in these genes occur at the DNA level, the methods described herein can be based on detecting mutations or variations in genomic DNA as well as transcripts and proteins themselves. It may be desirable to confirm mutations in genomic DNA by analyzing transcripts and / or polypeptides to ensure that the identified mutation is in fact expressed in the subject.

One skilled in the art will appreciate that there are a large number of analytical procedures that can be used to detect the presence or absence of nucleotide variants at one or more positions in a gene. Typically, detection of allelic variation requires a mutation discrimination technique, optionally an amplification reaction (such as based on polymerase chain reaction) and optionally a signal generating system. There are a large number of mutation detection techniques available in the art and can be used in combination with a signal generating system, many of which are available in the art. A review of the many methods for detecting allelic variation is given in Nollau et al., Clin. Chem. 1997, 43, 1114-1120; Anderson SM. Expert Rev Mol Diagn .. 2011, 1L 635-642; Meyerson M. et al., Nat Rev Genet., 2010, JJ, 685-696; and in standard guidelines such as Laboratory Protocols for Mutation Detection, ed. U. Landegren, Oxford University Press, 1996, and PCR, 2nd ed., Newton & Graham, BIOS Scientific Publishers Limited, 1997.

As indicated above, the determination of the presence or absence of a particular variation or multiple variations in the ERa gene in a patient suffering from cancer can be performed in various ways. Such tests are usually performed using DNA or RNA collected from biological samples, for example, biopsies of tissues, urine, feces, sputum, blood, cells, tissue scrapings, breast punctates, or other cellular materials, and can be performed in a variety of ways, including without limitations of PNR, hybridization with allele-specific probes, enzymatic detection of mutations, chemical cleavage of mismatch sites, mass spectrometry or DNA sequencing, including mini-sequencing.

Suitable mutation detection techniques include Amplification System for Mutation Identification (ARMS ™), Amplification System for Linear Extension Mutations (ALEX ™), Competitive Oligonucleotide Priming System (COPS), Taqman, molecular beacons, restriction fragment length polymorphism (RFLP) ), as well as PNR with the introduction of restriction sites and fluorescence resonance energy transfer (FRET).

In specific embodiments, the method used to determine the nucleotide (nucleotides) in the biomarker gene is selected from allele-specific amplification (allele-specific PCR), such as amplification system for mutation identification (ARMS), sequencing, allelic discriminant analysis, hybridization, restriction length polymorphism fragments (RFLP) or analysis by the method of ligation of oligonucleotide probes (OLA).

In certain embodiments, hybridization to allele-specific probes can be performed using (1) allele-specific oligonucleotides bound to a solid phase (e.g. glass, silicon, nylon membranes) in a labeled probe solution, such as in many DNA applications -chips; or (2) a linked sample (often cloned DNA or DNA amplified with PCR) in solution with labeled oligonucleotides (or allele

- 45 037533 specific, or short to ensure sequencing by hybridization). A set of variations can be included in diagnostic tests, often on a solid support, allowing more than one variation to be detected at a time. Such hybridization probes are well known in the art (see, for example, Green & Sambrook, Eds., Molecular Cloning: A Laboratory Manual, (2012, 4th ed., Volume 1-3, ISBN 9781936113422), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York) and may span two or more sites with variations.

Thus, in one embodiment, detecting the presence or absence of at least one mutation comprises contacting an ERa nucleic acid containing the putative mutation site with at least one nucleic acid probe. Under selective hybridization conditions, the probe preferably hybridizes to a nucleic acid sequence containing a site of variation and containing complementary nucleotide bases at a site of variation. Hybridization can be detected with a detectable label using labels known to a person skilled in the art. Such labels include, but are not limited to, radioactive, fluorescent, staining, and enzymatic labels.

In another embodiment, detecting the presence or absence of at least one mutation comprises contacting an ERa nucleic acid containing the putative mutation site with at least one nucleic acid primer. Under selective hybridization conditions, the primer preferably hybridizes to a nucleic acid sequence containing a site of variation and containing complementary nucleotide bases at a site of variation.

Oligonucleotides used as primers for specific amplification may contain a nucleotide base complementary to the mutation in question at the center of the molecule (such that amplification is dependent on differential hybridization; see, for example, Gibbs, et al., 1989. Nucl. Acids Res., 17, 2437-248) or at the very edge of the 3'-end of one primer, where, under appropriate conditions, mismatch of bases can prevent or reduce polymerase-mediated chain elongation (see, for example, Prossner, 1993, Tibtech. 11 238) ...

In yet another embodiment, detecting the presence or absence of at least one mutation comprises sequencing at least one nucleic acid sequence and comparing the resulting sequence to a known wild-type nucleic acid sequence.

Alternatively, the presence or absence of at least one mutation provides for the determination of the sequence of at least one nucleic acid using mass spectrometry.

In one embodiment, detecting the presence or absence of at least one variation in a nucleic acid involves performing a polymerase chain reaction (PCR). The target nucleic acid sequence containing the intended variation is amplified, and the nucleotide sequence of the amplified nucleic acid is determined. Determining the nucleotide sequence of the amplified nucleic acid involves sequencing at least one segment of the nucleic acid. Alternatively, the amplification products can be analyzed by any method suitable for separating amplification products according to their size, including automatic and manual gel electrophoresis and the like.

Mutations in genomic nucleic acid are predominantly detected using techniques based on changes in the mobility of amplified nucleic acid fragments. For example, Chen et al., Anal Biochem 1996, 239, 61-9, describe the detection of single base mutations by competitive mobility analysis. In addition, assay systems based on the method of Marcelino et al., BioTechniques 1999, 26, 1134-1148 are commercially available.

In a specific example, capillary heteroduplex (hybridization) analysis based on changes in the mobility of duplex nucleic acids in capillary systems as a result of base mismatch can be used to detect mutations.

To obtain nucleic acids from samples for analysis, amplification of nucleic acids is usually required. Many amplification methods rely on an enzymatic chain reaction (such as polymerase chain reaction, ligase chain reaction, or self-sustaining sequence replication) or on the replication of all or part of the vector into which it has been cloned. Preferably, the amplification in accordance with the present disclosure is exponential amplification, an example of which is the polymerase chain reaction.

The literature describes many methods for amplifying targets and signals, for example, general reviews of these methods are given in Landegren, U., et al., Science, 1988, 242, 229-237 and Lewis, R., Genetic Engineering News, 1990, 10. 54-55. Such amplification methods can be used in the methods according to the description provided by the present inventors, and they include polymerase chain reaction (PCR), in situ PCR, ligase amplification reaction (LAR), hybridization with ligase, Qe bacteriophage replicase, transcriptional mediated amplification system (TAS), gen- 46 037533 nominal amplification with transcript sequencing (GAWTS), nucleic acid sequence-based amplification (NASBA), and in situ hybridization. Primers suitable for use in various amplification techniques can be prepared according to methods known in the art.

Polymerase chain reaction (PCR) is a nucleic acid amplification technique described, inter alia, in US Pat. Nos. 4,683,195 and 4,683,202. PCR consists of repeated cycles of primer extension reactions caused by DNA polymerase. The target DNA is heat denatured and two oligonucleotides located at both ends of the target sequence on opposite strands of the DNA to be amplified hybridize. These oligonucleotides become primers for use with DNA polymerase. DNA is copied by extending the primer to produce a second copy of both strands. By repeating a cycle of heat denaturation, hybridization and primer extension, the target DNA can be amplified 1 million or more times in approximately 2-4 hours. PCR is a molecular biology tool that must be used in conjunction with detection techniques to determine the amplification results. The advantage of PCR is that it increases sensitivity by amplifying the number of target DNA by 1 million to 1 billion times over about 4 hours. PCR can be used to amplify any known nucleic acid for diagnostic purposes (Mok et al., Gvnaecologic Oncology. 1994 , 52: 247-252).

Allele-specific amplification techniques such as the Amplification System for Mutation Identification (ARMS ™) (Newton et al., Nucleic Acids Res., 1989, 17, 2503-2516) can also be used to detect single base mutations. Under appropriate conditions for PCR amplification, for the preferred amplification of perfectly matching alleles (Newton et al., 1989, above), a base pair mismatch occurring at the 3'-end of the primer is sufficient to distinguish closely related molecules. The amplification system, which uses the primers described above, is based on the fact that oligonucleotides with mismatched residues at the 3 'end will not function as primers for PCR under appropriate conditions. This amplification system allows genotyping solely by examining reaction mixtures after agarose gel electrophoresis.

Analysis of amplification products can be performed using any method capable of separating amplification products according to their size, including automatic and manual gel electrophoresis, mass spectrometry, and the like.

Methods for isolating, amplifying, and analyzing nucleic acid are common to those of skill in the art, and examples of protocols can be found, for example, in Green & Sambrook, Eds., Molecular Cloning: A Laboratory Manual, (2012, 4th ed., Vol. 1-3, ISBN 9781936113422), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) A particularly useful source of protocols for methods used in PCR amplification is PCR (Basics: From Background to Bench), MJ McPherson, S.G. Mailer, R. Beynon, C. Howe, Springer Verlag; 1st ed. (October 15, 2000), ISBN: 0387916008.

Also provided herein are predictive and diagnostic kits containing degenerate primers for amplifying a target nucleic acid sequence from the ERa gene and instructions including an amplification protocol and analysis of the results. Alternatively, the kit may also contain buffers, enzymes, and containers for performing amplification and analyzing amplification products. The kit can also be a component of a screening or diagnostic kit containing other tools such as DNA microarrays or other supports. Preferably, the kit also provides one or more templates with controls such as nucleic acids isolated from normal tissue samples and / or rows of samples representing different variations in reference genes.

In one embodiment, the kit includes two or more pairs of primers, each pair being suitable for amplifying a separate region of a reference gene (ERa) (each region is a site of possible variation), thereby providing a kit for assay in a biological sample for expression of multiple variations in a gene in one reaction or in several parallel reactions.

The primers in the kits can be labeled, eg, fluorescently labeled, to aid in the detection of amplification products and subsequent analysis of variations in the nucleic acid. The kit can also be designed to detect more than one variation at a time. Therefore, the combination kit will contain primers suitable for amplifying different segments of the reference gene. Primers can be labeled differentially, for example using different fluorescent labels, to differentiate between different variations.

In another aspect, provided herein is a method of treating a patient suffering from cancer, comprising assigning the ERa gene in the patient's tumor cells the status of a mutant gene or a wild-type gene and, if the ERa gene is mutant, administering to the patient an effective amount of the compound of formula (ID) ...

As used herein, the terms effective and efficacy encompass both pharmacological efficacy and physiological safety. Pharmacological efficacy refers to the ability of a treatment to provide a desired biological effect in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects at the cellular, organ and / or body level (often referred to as side effects) resulting from the administration of a therapeutic agent. Less effective means that the treatment results in a therapeutically less significant level of pharmacological efficacy and / or a therapeutically higher level of adverse physiological effects.

In accordance with another aspect of the present disclosure, there is provided the use of a compound of formula (ID), or a pharmaceutically acceptable salt thereof, for treating a patient suffering from cancer whose tumor cells have been identified as containing a mutant ERa gene.

In accordance with another aspect of the present disclosure, there is provided a compound of formula (ID), or a pharmaceutically acceptable salt thereof, for the treatment of cancers in which tumor cells have been identified as carrying the mutant ERa gene.

In accordance with another aspect of the present disclosure, there is provided a method of treating cancers in which tumor cells are identified as carrying the mutant ERa gene, comprising administering an effective amount of a compound of formula (ID), or a pharmaceutically acceptable salt thereof.

In still several additional embodiments, the implementation of the present description relates to a pharmaceutical composition containing a compound of formula (ID), intended for use in the prevention and treatment of cancer with tumor cells identified as carrying the mutant ERa gene.

For all of the above aspects, detectable / detectable mutant forms of ERa are those at any position throughout the gene.

For all of the above aspects, when used as an example of a tumor such as breast cancer, specific identified / identified ERa mutants are those with mutations at Ser463Pro, Val543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn and Asp538Gly.

Description of graphic materials

FIG. 1 shows the X-ray powder diffraction pattern for Form A N- (1- (3-fluoropropyl) azetidin-3-yl) 6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine-3-amine.

FIG. 2 shows the DSC / TGA thermogram for Form A N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine.

FIG. 3 shows the X-ray powder diffraction pattern for Form B N- (1- (3-fluoropropyl) azetidin-3-yl) 6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine-3-amine.

FIG. 4 shows the X-ray powder diffraction pattern for form C N- (1- (3-fluoropropyl) azetidin-3-yl) 6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine-3-amine.

FIG. 5 shows the DSC / TGA thermogram for Form C N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine.

FIG. 6 shows the X-ray powder diffraction pattern for Form D N- (1- (3-fluoropropyl) azetidin-3-yl) 6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine-3-amine.

FIG. 7 shows the DSC / TGA thermogram for Form D N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine.

FIG. 8 shows the X-ray powder diffraction pattern for Form E N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine.

FIG. 9 shows the DSC / TGA thermogram for Form E N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine.

FIG. 10 shows the X-ray powder diffraction pattern for Form F N- (1- (3-fluoropropyl) azetidin-3-yl) 6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine-3-amine.

FIG. 11 shows the X-ray powder diffraction pattern for Form G N- (1- (3-fluoropropyl) azetidin-3-yl)

- 48 037533

6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) pyridine -3 -amine.

FIG. 12 shows the results of a study on a mouse model of the antitumor efficacy of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine in relation to the xenograft representing the original human MCF7.

FIG. 13 shows the results of a study on a mouse model of the antitumor efficacy of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine in relation to the human xenograft, which is MCF7 with the Y537S mutation in ESR1.

FIG. 14 shows the results of a study on a mouse model of the antitumor efficacy of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine in relation to a human xenograft, which is CTC174, obtained from a patient with breast cancer and mutation in ESR1 ...

FIG. 15 shows the results of a mouse model study of the antitumor efficacy of a combination of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl ) -6,7,8,9tetrahydro-3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridine-3-amine and palbociclib in relation to a human xenograft, which is CTC174, obtained from a patient with breast cancer and mutation in ESR1.

FIG. 16 shows the results of a study on a mouse model of the antitumor efficacy of a combination of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl ) -6,7,8,9tetrahydro-3H-pyrazolo [4,3- £] isoquinolin-6-yl) pyridin-3-amine and visusertib (AZD2014) in relation to a human xenograft, which is CTC174, obtained from a patient with cancer mammary gland and mutation in ESR1.

Examples of

The compounds described herein are further illustrated in the following examples. These examples are given by way of illustration and not limitation. In general, the following is envisaged.

(i) Operations were carried out at ambient temperature, i. e. in the range of 17-25 ° C and in an atmosphere of an inert gas such as nitrogen, unless otherwise indicated.

(ii) Evaporation was carried out by rotary evaporation or using Genevac equipment or a Biotage v10 in vacuo evaporator, and product isolation procedures were performed after removal of residual solids by filtration.

(iii) Purification by flash chromatography was performed on a Teledyne Isco CombiFlash® Rf automated system or Teledyne Isco CombiFlash® Companion® using pre-packed RediSep Rf Gold ™ columns (20-40 μm, spherical particles), GraceResolv ™ cartridges (Davisil silica gel ®) or Silicycle cartridges (40-63 μm).

(iv) Preparative chromatography was performed on a Gilson preparative HPLC with UV collection system or by supercritical liquid chromatography performed on a Waters Prep 100 SFC-MS device with MS and UV collection systems or a Thar MultiGram III SFC with UV collection system. collection system.

(v) Chiral preparative chromatography was performed on a Gilson UV collection system (233 injector / fraction collector, 333 and 334 pumps, 155 UV detector) or on a Varian Prep Star device (2x SD1 pumps, 325 UV detector, 701 collectors fractions) with a pump working with a Gilson 305 injector.

(vi) Yield values, if any, were not necessarily the maximum attainable.

(vii) Typically, the structures of the final products of formula I were confirmed by nuclear magnetic resonance (NMR) spectroscopy; NMR chemical shift values were measured on a delt-scale [proton magnetic resonance spectra were determined using a Bruker Avance 500 (500 MHz) or Bruker Avance 400 (400 MHz)]; measurements were carried out at ambient temperature, unless otherwise indicated; 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 doublets; dt, doublet of triplets; bs, wide signal.

(viii) Typically, the final products of Formula I were also characterized by liquid chromatography mass spectroscopy (LCMS or UPLC); UPLC was performed using a Waters UPLC equipped with a Waters SQ mass spectrometer (column temperature 40, UV = 220-300 nm, mass spectrometry = ESI with positive / negative charge switching) at a flow rate of 1 ml / min, using a solvent system from 97% A + 3% B up to 3% A up to 97% B in 1.50 min (total run time with equilibration to baseline, etc. 1.70 min.), Where A = 0.1% formic acid in water (for working with acid) or 0.1% ammonia in water (for working with a base); B = acetonitrile. For acid analysis, the column used was Waters Acquity HSS T3, 1.8

- 49 037533 µm, 2.1x50 mm, for the main analysis the column used was a Waters Acquity BEN 1.7 µm, 2.1x50 mm; LCMS was performed using a Waters Alliance HT (2795) system equipped with a Waters ZQ ESCi mass spectrometer and a Phenomenex Gemini-NX column (50x2.1 mm, 5 μm) at a flow rate of 1.1 ml / min from 95% A to 95% B in 4 min with a hold of 0.5 min. The modifier was kept constant at 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 the method is acidic or basic.

(ix) Ion exchange purification was typically performed using an SCX-2 cartridge (Biotage, propyl sulfonic acid functionalized silica. Manufactured using a trifunctional silane. No end group blocking).

(x) The purity of the intermediate was assessed by thin layer chromatography, mass spectrometric, HPLC (high performance liquid chromatography) and / or NMR analysis.

(xi) The following abbreviations were used:

Ason acetic acid aq. water Vos tert-butoxycarbonyl n-BuLi n-butyllithium tBuOH tert-butanol Brettphos 2- (dicyclohexylphosphino) 3,6-dimethoxy- 2 ', 4', 6 '-triisopropyl-1,1' -biphenyl CDC1 ₃ deutero-chloroform CHC1 ₃ chloroform Conc. concentrated DCM dichloromethane DIPEA diisopropylethylamine DMAP dimethylaminopyridine DMSO dimethyl sulfoxide EtOH ethanol EtOAc ethyl acetate HATU hexafluorophosphate 1-

- 50 037533

[bis (dimethylamino) methylene] -1 // - 1,2,3triazolo [4,5 -th] pyridinium-3-oxide HC1 hydrochloric acid HPLC high performance liquid chromatography K ₂ CO ₃ potassium carbonate Meon methanol MgSO ₄ magnesium sulfate NaH sodium hydride NaHCO ₃ bicarbonate of soda Na ₂ SO ₄ sodium sulfate Pd ₂ (dba) ₃ tris (dibeisylideneacetone) dipalladium (0) Rac-BINAP (±) -2,2'-Bis (diphenylphosphino) -1, GBinaphthalene K. T. room temperature prekatalizator ^3rd [(2-Di-tert-butylphosphino-3-methoxy-6- generation RockPhos methyl-2 ', 46' -triisopropyl-1,1 'biphenyl) -2- (2- aminobifenyl)] palladium (P) methanesulfonate Ruphos 2-Dicyclohexylphosphino-2 ', 6' diisopropoxy-1,1 '-biphenyl saturate saturated SFC supercritical liquid chromatography sol. solution TBAF tetra-n-butylammonium fluoride TEA triethylamine THF tetrahydrofuran THP tetrahydropyran TFA trifluoroacetic acid Xantphos 4,5-bis (diphenylphosphino) -9,9-dimethyl

Example 1.

N- (4 - ((6S, 8R) -7- (2,2-Difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline- 6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine

[(2-Dicyclohexylphosphino-3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'biphenyl)] palladium (11) methanesulfonate (BrettPhos Pd G3) (6.80 mg, 7.50 μmol) was added to a suspension of (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl ) -8-methyl-3- (tetrahydro-2H-pyran-2yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (85 mg, 0.15 mmol), 1 - (3-fluoropropyl) azetidine-3amine (29.7 mg, 0.23 mmol) and sodium tert-butoxide (28.8 mg, 0.30 mmol) in degassed 1,4-dioxane (1.5 ml) and the reaction mixture was heated to 90 ° C for 3 hours. After cooling, the reaction mixture

- 51 037533 was diluted with DCM and washed with water. The aqueous layer was extracted with DCM, then the combined organics were dried and evaporated. The crude residue was dissolved in DCM (2 ml) and TFA (0.7 ml) was added. The reaction mixture was stirred at room temperature for 1 hour, then it was diluted with DCM and made basic by the addition of saturated NaHCO ₃ solution. The layers were separated and the aqueous layer was extracted with DCM. The combined organics were dried and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in EtOAc. Pure fractions were evaporated to dryness to give N- (4 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine (39.0 mg, 49%) as a beige solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.09 (3H, d), 1.83 (2H, ddd), 2.65-2.86 (4H, m), 2.96-3, 08 (1H, m), 3.11 (1H, dd), 3.22 (2H, d), 3.38 (3H, s), 3.51-3.63 (2H, m), 3.73 -3.82 (1H, m), 3.82 (3H, s), 3.86 (2H, q), 4.15-4.28 (1H, m), 4.43 (1H, t), 4.53 (1H, t), 5.37 (1H, s), 5.88 (1H, dd), 6.14 (1H, d), 6.46 (1H, d), 6.80 (1H , d), 7.13 (1H, d), 8.04 (1H, d);

mass / charge: ES + [M + H] + = 532.

(6S, 8R) -6- (4-Bromo-2-methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline was prepared as follows.

Preparation of 2,2-difluoro-3- (trityloxy) propan-1-ol

I ^Ph _vPh HO ^^^^ CT ^ Ph

F F

2,2-Difluoropropane-1,3-diol (2.50 g, 22.3 mmol) was dissolved in DCM (61.7 ml) and THF (15.4 ml). DIPEA (3.93 mL, 22.3 mmol) was added followed by the addition of (chloromethanetriyl) tribenzene (6.22 g, 22.3 mmol) and finally DMAP (0.288 g, 2.23 mmol). The reaction mixture was heated to 40 ° C for 2 h. After cooling, the reaction mixture was washed with 1N. HCl solution, then dried and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 25% EtOAc in heptane. The pure fractions were evaporated to dryness to give 2,2-difluoro-3- (trityloxy) propan-1-ol (4.43 g, 56%) as a colorless solid.

1H NMR (500 MHz, CDCl3) 3.42 (2H, t), 3.92 (2H, t), 7.23-7.3 (4H, m), 7.3-7.39 (6H, m ), 7.39-7.49 (6H, m).

Obtaining ((2,2-difluoro-3-methoxypropoxy) methanetriyl) tribenzene ^Pt ! Ph

Ph

F F

Sodium hydride (0.562 g, 14.0 mmol) was added to a solution of 2,2-difluoro-3- (trityloxy) propan-1-ol (4.15 g, 11.7 mmol) in THF (46 ml) and the reaction mixture stirred for 1 h then iodomethane (0.802 ml, 12.9 mmol) in THF (5 ml) was added. The reaction mixture was stirred for an additional 1 h. The reaction mixture was quenched with water and brine, then extracted with EtOAc. The organic layer was dried and evaporated to give ((2,2-difluoro-3methoxypropoxy) methanetriyl) tribenzene (4.18 g, 97%) as a pale yellow oil.

1H NMR (500 MHz, CDCl ₃ ) 3.36 (2H, t), 3.40 (3H, s), 3.77 (2H, t), 7.15-7.28 (3H, m), 7 , 28-7.38 (6H, m), 7.39-7.47 (6H, m).

Preparation of 2,2-difluoro-3-methoxypropyl trifluoromethanesulfonate o L ο ^ ο% ^F

Trifluoromethanesulfonic anhydride (1.918 ml, 11.40 mmol) was added to a solution of ((2,2-difluoro-3-methoxypropoxy) methanetriyl) tribenzene (4.00 g, 10.9 mmol) in DCM (39.6 ml). The reaction mixture was stirred for 30 min, then triethylsilane (1.934 ml, 11.94 mmol) was added and the reaction mixture was stirred for an additional 30 min. The reaction mixture was evaporated and the triflate was used directly in the next step.

Obtaining (R) -1- (3-bromo-2-methylphenyl) propan-2-amine

n-BuLi (26.1 ml, 41.8 mmol) was added dropwise to a solution of 1,3-dibromo-2-methylbenzene (9.95 g, 39.8 mmol) in THF (100 ml) at -78 ° C. ... After stirring for 30 min, (R) -tert-butyl-4-methyl-1,2,3-oxathiazolidine-3-carboxylate-2,2-dioxide (10.39 g, 43.8 mmol) was added in portions and the reaction the mixture was stirred for an additional 30 minutes before allowing it to warm to 0 ° C over 30 minutes. Added 1N. citric acid and the mixture was stirred for

52,037533 min before it was extracted with EtOAc (x2). The combined organic phases were evaporated. The residue was stirred in 4M HCl in dioxane (69.7 mL, 278.7 mmol) at room temperature for 1 hour, then the volatiles were evaporated. The residue was suspended in diethyl ether and extracted with water (x2). The combined aqueous phases were podslushivaet by adding 2N. Na ₂ CO ₃ , then extracted with DCM (x3). The combined organic phases were dried over MgSO ₄ and concentrated to give (R) -1- (3-bromo-2-methylphenyl) propan-2-amine (7.55 g, 83%) as a yellow oil.

'and NMR (400 MHz, CDCl3, 27 ° С) 1.13 (3H, d), 1.43 (2H, s), 2.40 (3H, s), 2.61 (1H, dd), 2 , 77 (1H, dd), 3.14 (1H, dq), 6.97 (1H, t), 7.08 (1H, d), 7.43 (1H, d);

mass / charge (ES ⁺ ) [M + H] ⁺ = 228.

Preparation of (R) -N- (1- (3-bromo-2-methylphenyl) propan-2-yl) -2,2-difluoro-3-methoxypropan-1-amine

2,2-Difluoro-3-methoxypropyl trifluoromethanesulfonate (3.03 g, 11.73 mmol) (crude, from the previous step) was added to a solution of (R) -1- (3-bromo-2-methylphenyl) propan-2-amine (2.327 g, 10.2 mmol) and DIPEA (2.82 ml, 16.32 mmol) in 1,4-dioxane (34.3 ml) and the reaction mixture was heated to 80 ° C overnight. After cooling, the volatiles were evaporated, then the residue was taken up in DCM and washed with brine. The organic phase was dried and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 100% EtOAc in heptane. Pure fractions were evaporated to dryness to give (R) -N- (1- (3-bromo-2-methylphenyl) propan-2-yl) -2,2-difluoro-3-methoxypropan-1-amine (2.330 g, 68%) as a pale yellow oil.

1H NMR (500 MHz, CDCl ₃ ) 1.06 (3H, d), 2.40 (3H, s), 2.63 (1H, dd), 2.82 (1H, dd), 2.86-2 , 94 (1H, m), 2.95-3.11 (2H, m), 3.38 (3H, s), 3.51-3.63 (2H, m), 6.94-7 (1H , m), 7.07 (1H, dd), 7.4-7.51 (1H, m);

mass / charge: ES ⁺ [M + H] ⁺ = 336.

Preparation of (R) -3- (2 - ((2,2-difluoro-3-methoxypropyl) amino) propyl) -2-methylaniline

Pd ₂ (dba) ₃ (0.184 g, 0.20 mmol) and Rac-BINAP (0.250 g, 0.40 mmol) were added to a suspension of (R) -N- (1 (3-bromo-2-methylphenyl) propane- 2-yl) -2,2-difluoro-3-methoxypropan-1-amine (2.25 g, 6.69 mmol), benzophenone imine (1.334 g, 7.36 mmol) and sodium tert-butoxide (0.965 g, 10 , 0 mmol) in degassed toluene (28.5 ml) and the reaction mixture was heated to 90 ° C for 3 h. After cooling, a significant amount of toluene was evaporated, then the residue was dissolved in DCM and washed with water. The aqueous phase was extracted with DCM, then the organics were evaporated to a volume of ~ 50 ml. Added 2N. HCl solution (50 ml) and the biphasic mixture were vigorously stirred for 30 min. The layers were separated, then the aqueous layer was extracted with DCM. The organic phase was again extracted with 1N. HCl. The combined aqueous phases were podslushivaet by adding solid K ₂ CO ₃ , then extracted with DCM (x3) and the combined extracts in DCM were dried and evaporated to give (R) -3- (2 - ((2,2-difluoro-3-methoxypropyl ) amino) propyl) -2-methylaniline (1.780 g, 98%) as a light brown oil.

1H NMR (500 MHz, CDCl ₃ ) 1.06 (3H, d), 2.11 (3H, s), 2.59 (1H, dd), 2.75 (1H, dd), 2.86-2 , 94 (1H, m), 2.94-3.12 (2H, m), 3.38 (3H, s), 3.53-3.63 (4H, m), 6.5-6.68 (2H, m), 6.86-7.01 (1H, m);

mass / charge: ES ⁺ [M + H] ⁺ 273.

Preparation of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2,2-difluoro-3-methoxypropyl) -3,5-dimethyl

1,2,3,4-tetrahydroisoquinoline-6-amine

(R) -3- (2 - ((2,2-Difluoro-3-methoxypropyl) amino) propyl) -2-methylaniline (490 mg, 1.80 mmol) and 4-bromo-2-methoxybenzaldehyde (813 mg, 3.78 mmol) was heated in acetic acid (8.8 ml) and water (0.162 ml, 9.00 mmol) to 75 ° C overnight. After cooling, the acetic acid was evaporated in vacuo, then the residue was dissolved in EtOAc (20 ml) and 2N. HCl solution (20 ml). The biphasic mixture was stirred for 30 min, then the layers were separated. The organic phase was extracted with water, then the aqueous phase was podslushivaet by adding 2N. NaOH solution and extracted with DCM (x2). The combined DCM layers were dried and evaporated, then the crude pro 53 037533 duct was purified by flash chromatography on silica, elution gradient from 0 to

50% EtOAc in heptane. Pure fractions were evaporated to dryness to give (1S, 3R) -1- (4-6pom2-methoxyphenyl) -2- (2,2-difluoro-3-methoxypropyl) -3,5-dimethyl-1,2,3, 4-tetrahydroisoquinoline-6-amine (384 mg, 46%) as a beige solid.

1H NMR (500 MHz, CDCR 27 ° С) 1.03 (3H, d), 2.07 (3H, s), 2.42 (1H, dd), 2.60-2.76 (2H, m) , 2.99 (1H, ddd), 3.34 (1H, d), 3.36 (3H, s), 3.52 (2H, s), 3.54-3.66 (1H, m), 3.76 (1H, ddd), 3.87 (3H, s), 5.28 (1H, s), 6.47 (2H, s), 6.59 (1H, d), 6.88 (1H , dd), 7.01 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 469.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4 , 3-f] isoquinoline

Sodium nitrite (59.8 mg, 0.87 mmol) was added in water (0.2 ml) to a cooled solution of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2.2- difluoro-3-methoxypropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (370 mg, 0.79 mmol) in propionic acid (2628 μl) / water (526 μl) at - 10 ° C. The reaction mixture was stirred for 1 h, then ice-cold EtOAc (20 ml) was added. The reaction mixture was quenched by the addition of cold saturated NaHCO ₃ solution and stirred for 15 min before being allowed to warm to room temperature. The layers were separated and the aqueous layer was extracted with EtOAc. The combined organics were dried and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane. Pure fractions were evaporated to dryness to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8, 9-tetrahydro3H-pyrazolo [4,3-f] isoquinoline (252 mg, 67%) as a beige solid.

¹ H NMR (500 MHz, CDCR 27 ° C) 1.10 (3H, d), 2.72 (1H, td), 2.85 (1H, dd), 3.08 (1H, ddd), 3, 16 (1H, dd), 3.36 (3H, s), 3.48 - 3.66 (2H, m), 3.65- 3.81 (1H, m), 3.90 (3H, s) , 5.44 (1H, s), 6.59 (1H, d), 6.77 (1H, d), 6.89 (1H, dd), 7.05 (1H, d), 7.19 ( 1H, dd), 8.07 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 480.

Preparation of (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline Br j0 jCU ^ ⁰

3,4-Dihydro-2H-pyran (0.064 ml, 0.70 mmol) was added to a solution of (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (240 mg, 0.50 mmol) and p-toluenesulfonic acid hydrate (9.51 mg, 0.05 mmol) in DCM (2.5 ml) and the reaction mixture was heated to 40 ° C for 1 h. After cooling, the reaction mixture was diluted with DCM and washed with saturated NaHCO ₃ solution. The organic phase was dried and evaporated. The crude residue was passed through a silica plug (EtOAc / heptane, 1: 1, as eluent) and the filtrate was evaporated to give (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7- (2,2-difluoro- 3-methoxypropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9 tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (262 mg, 93%) in as a pale yellow solid as a 5: 1 ratio of THP regioisomers (each THP regioisomer is a diastereoisomeric mixture).

Mass / charge: ES ⁺ [M + H] ⁺ = 564.

Preparation of tert-butyl (1 - (3-fluoropropyl) azetidin-3-yl) carbamate

Potassium carbonate (1.605 g, 11.61 mmol), tert-butylazetidin-3-yl carbamate (1.0 g, 5.81 mmol) and

- 54 037533

1-fluoro-3-iodopropane (1.146 g, 6.10 mmol) was suspended in acetonitrile (11.61 ml) and sealed in a microwave tube. The reaction mixture was heated to 95 ° C for 15 minutes in a microwave reactor. The reaction mixture was cooled to room temperature, diluted with EtOAc (100 ml) and extracted from saturated NaHCO ₃ (50 ml). The organic phase was dried over MgSO ₄ , filtered and evaporated to give tert-butyl (1- (3fluoropropyl) azetidin-3-yl) carbamate (1.248 g, 93%) as a yellow solid.

¹ H NMR (500 MHz, CDCl3, 22 ° C) 1.44 (9H, s), 1.68-1.80 (2H, m), 2.56 (2H, t), 2.88 (2H, s), 3.66 (2H, t), 4.31 (1H, d), 4.43 (1H, t), 4.53 (1H, t), 4.90 (1H, s).

Obtaining 1- (3-fluoropropyl) azetidine-3-amine nh ₂

TFA (2.69 ml) was added to a solution of tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate (1.248 g, 5.37 mmol) in DCM (8.06 ml) and the reaction mixture was stirred at room temperature for 1 hour. Volatiles were evaporated and purified by ion exchange chromatography using an SCX column. The desired product was eluted from the column using 1M NH ₃ / MeOH and the pure fractions were evaporated to dryness to give 1- (3-fluoropropyl) azetidine-3-amine (0.697 g, 98%) as a colorless oil.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.66-1.83 (2H, m), 1.96 (3H, s), 2.54 (2H, t), 2.67 (2H, td ), 3.58-3.70 (2H, m), 4.43 (1H, t), 4.52 (1H, t).

Example 2.

6 - ((6S, 8R) -7- (2,2-Difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

[(2-Dicyclohexylphosphino-3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'biphenyl)] palladium (P) methanesulfonate (BrettPhos Pd G3) (6.83 mg, 8.00 μmol) and sodium tert-butoxide (48.0 mg, 0.50 mmol) were added to a degassed solution of (6S, 8R) -6- (5-bromopyridine 2-ul) -7- (2,2difluoro-3-methoxypropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4 , 3f] isoquinoline (107 mg, 0.20 mmol) and 1- (3-fluoropropyl) azetidine-3-amine (52.9 mg, 0.40 mmol) in 1,4-dioxane (1.6 ml) and the reaction mixture was heated to 90 ° C for 5 h. After cooling, the reaction mixture was diluted with DCM and washed with water. The organic phase was evaporated, then dissolved in DCM (2 ml) before TFA (1 ml) was added. The mixture was stirred at room temperature for 1 h, then diluted with DCM and washed with saturated NaHCO ₃ solution. The layers were separated and the aqueous layer was extracted with DCM. The combined organics were dried and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in EtOAc. Pure fractions were evaporated to dryness to give 6 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4.3 -f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3yl) pyridin-3-amine (83 mg, 83%) as a beige solid.

1H NMR (500 MHz, CDCl ₃ ) 1.12 (3H, d), 1.79 (2H, ddd), 2.71 (2H, t), 2.75-2.90 (2H, m), 3 , 06-3.23 (3H, m), 3.39 (3H, s), 3.51- 3.66 (2H, m), 3.66-3.76 (1H, m), 3.76 -3.84 (2H, m), 4.14 (1H, s), 4.44 (2H, t), 4.53 (1H, t), 5.06 (1H, s), 6.80 ( 1H, dd), 6.88 (1H, d), 7.13 (2H, dd), 7.84 (1H, d), 7.99 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 503.

(6S, 8R) -6- (5-Bromopyridin-2-yl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-3- (tetrahydro-2H-pyran2-yl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline was prepared as follows.

Preparation of (1S, 3R) -1- (5-bromopyrudin-2-yl) -2- (2,2-difluoro-3-methoxypropyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine

Br

5-Bromopicolinaldehyde (1172 mg, 6.30 mmol) was added to a solution of (R) -3- (2 - ((2,2-difluoro-3

- 55 037533 methoxypropyl) amino) propyl) -2-methylaniline (817 mg, 3.00 mmol) in acetic acid (14.7 ml) and water (270 μl, 15.0 mmol) and the reaction mixture was heated to 80 ° C within 2 hours. After cooling, the volatiles were evaporated in vacuo. The residue was dissolved in DCM and washed with saturated NaHCO ₃ solution. The organic matter was evaporated to a volume of ~ 20 ml and 2N. HCl solution (20 ml). The biphasic mixture was stirred for 15 minutes, then separated. The organic matter was extracted with water, then the aqueous phase was again extracted with DCM. The aqueous phase was then podslushivaet by adding solid K ₂ CO ₃ , then extracted with DCM. The organic extracts were dried and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane. Pure fractions were evaporated to dryness to give (1S, 3R) -1- (5-bromopyridin-2-yl) -2- (2,2-difluoro-3methoxypropyl) -3,5-dimethyl-1,2,3, 4-tetrahydroisoquinoline-6-amine (810 mg, 61%) as a beige solid.

¹ H NMR (500 MHz, CDCl ₃ ) 1.07 (3H, d), 2.05 (3H, s), 2.49 (1H, d), 2.75 (2H, dd), 3.04- 3.17 (1H, m), 3.30-3.36 (1H, m), 3.37 (3H, s), 3.58-3.74 (2H, m), 4.96 (1H, s), 6.51 (1H, d), 6.60 (1H, d), 7.22 (1H, d), 7.68 (1H, dd), 8.55 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 440.

Obtaining (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4 , 3-f] isoquinoline

Sodium nitrite (133 mg, 1.93 mmol) was added in water (0.5 ml) to a cooled solution of (1 S, 3R) -1 - (5-bromopyridin-2-yl) -2- (2,2-difluoro -3-methoxypropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (771 mg, 1.75 mmol) in propionic acid (5833 μl) / water (1167 μl) at -15 ° C. The reaction mixture was stirred for 30 min, then EtOAc (50 ml) was added, which was cooled in dry ice. The reaction mixture was quenched by the addition of 2N. Na ₂ CO ₃ until the evolution of gas bubbles stops, then the layers were separated. The aqueous layer was extracted with EtOAc, then the organic was dried and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane. Pure fractions were evaporated to dryness to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoro-3methoxypropyl) -8-methyl-6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (460 mg, 58%) as a pale yellow solid.

¹ H NMR (500 MHz, CDCl ₃ ) 1.14 (3H, d), 2.81 (1H, dd), 2.88 (1H, dd), 3.10-3.26 (2H, m), 3.38 (3H, s), 3.46-3.55 (1H, m), 3.58-3.76 (2H, m), 5.13 (1H, s), 6.94 (1H, d), 7.23 (1H, dd), 7.29 (1H, d), 7.72 (1H, dd), 8.05 (1H, d), 8.57 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 451.

Preparation of (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3] isoquinoline

3,4-Dihydro-2H-pyran (0.114 ml, 1.25 mmol) was added to a solution of (6S, 8R) -6- (5-bromopyridine-2yl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (450 mg, 1.00 mmol) and PTSA hydrate (37.9 mg, 0.20 mmol) in DCM (5 ml) and the reaction mixture was heated to 45 ° C for 3 h. After cooling, the reaction mixture was diluted with DCM and washed with saturated NaHCO ₃ solution. The organic phase was dried and evaporated, then the crude material was passed through a plug of silica (EtOAc / heptane 1: 1). The filtrate was evaporated to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl ) -6,7,8,9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinoline (510 mg, 9%) as an orange solid (~ 6.5: 1 ratio of THP regioisomers).

Mass / charge: ES ⁺ [M + H] ⁺ = 535.

- 56 037533

Example 3. 6 - ((6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3] isoquinolin-6-yl ) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

[(2-Dicyclohexylphosphino-3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'biphenyl)] palladium (11) methanesulfonate (BrettPhos Pd G3) (12.11 mg, 0.01 mmol) and sodium tert-butoxide (85 mg, 0.89 mmol) were added to a degassed solution of (6S, 8R) -6- (5-bromopyridine-2- yl) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline ( 177 mg, 0.35 mmol) and 1- (3-fluoropropyl) azetidin-3-amine (94 mg, 0.71 mmol) in 1,4 dioxane (2835 μL) and the reaction mixture was heated to 90 ° С for 5 h After cooling, the reaction mixture was diluted with DCM and washed with water. The organic phase was evaporated, then dissolved in DCM (2 ml) before TFA (1 ml) was added. The mixture was stirred at room temperature for 1 h, then diluted with DCM and washed with saturated NaHCO ₃ solution. The layers were separated and the aqueous phase was extracted with DCM. The combined organics were dried and evaporated to give the crude product. The crude product was purified by preparative LCMS (Waters SunFire column, 5 μm silica, 19 mm in diameter, 100 mm in length) using mixtures of water (containing 1% NH ₃ ) and MeCN with decreasing polarity as eluents. Fractions containing the required compounds were evaporated to dryness to obtain 6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrαzolo [4, 3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine (17.0 mg, 10%) as a beige solid.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.36-0.46 (1H, m), 0.49-0.59 (1H, m), 0.88-1.07 (2H, m), 1.09 (3H, d), 1.69-1.75 (1H, m), 1.75-1.81 (1H, m), 2.59 (2H, t), 2.70 (1H, dd), 2.86-2.96 (3H, m), 3.01-3.11 (1H, m), 3.373.44 (1H, m), 3.72 (2H, q), 3.78-3.88 (1H, m), 3.95 (1H, d), 4.04-4.13 (1H, m), 4.44 (1H, t), 4.53 (1H, t), 4.95 (1H, s), 6.75 (1H, dd), 6.91 (1H, d), 7.12-7.17 (2H, m), 7.86 (1H, d ), 8.05 (1H, d), 10.04 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 467.

(6S, 8R) -6- (5-Bromopyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline was prepared as follows.

Obtaining 1- (3-bromo-2-methylphenyl) -2,5-dimethyl-1H-pyrrole

A mixture of 3-bromo-2-methylaniline (40 g, 215 mmol), hexane-2,5-dione (25.3 ml, 215 mmol) and n-toluenesulfonic acid monohydrate (0.409 g, 2.15 mmol) in toluene ( 300 ml) was heated under reflux for 2 h in a flask equipped with a condenser and a Dean-Stark trap. The mixture was then cooled to room temperature and washed sequentially with saturated aqueous sodium bicarbonate solution, aqueous HCl (1N) and saturated aqueous sodium chloride solution. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude 1- (3-bromo-2-methylphenyl) -2,5-dimethyl-1H-pyrrole (57.9 g, 102%) as a pale yellow oil.

1H NMR (300 MHz, DMSO-d6, 27 ° С) 1.8 (6H, s), 1.9 (3H, s), 5.8 (2H, s), 7.2-7.4 (2H , m), 7.7 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 264.

Preparation of tert-butyl (R) - (1 - (3- (2,5-dimethyl-1 H-pyrrol-1-yl) -2-methylphenyl) propan-2yl) carbamate

n-Butyllithium in hexane (2.5 M; 89 ml, 221 mmol) was added over 15 min to a solution of crude 1- (3-bromo-2-methylphenyl) -2,5-dimethyl-1H-pyrrole (55.7 g, 211 mmol) in THF (400 ml) at -78 ° C. After 30 min, tert-butyl (R) -4-methyl-1,2,3-oxathiazolidine-3-carboxylate-2,2-dioxide (50 g, 211 mmol) was added. The resulting mixture was stirred at -78 ° C for 15 minutes and allowed to warm to room temperature over 2 hours. Aqueous citric acid solution (1N; 250 ml) was added and stirring was continued for 30 minutes. The mixture was extracted with hexanes

- 57 037533 and the combined organic layers were washed with saturated aqueous sodium carbonate solution, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting brown solid, crude tert-butyl (R) - (1- (3- (2,5-dimethyl-1H-pyrrol-1-yl) -2methylphenyl) propan-2-yl) carbamate, was used in the next step without additional cleaning.

Mass / charge: ES + [M + H] ⁺ = 343.

Preparation of (R) -1- (3- (2,5-dimethyl-1H-pyrrol-1-yl) -2-methylphenyl) propan-2-amine

Hydrochloric acid in dioxane (4 M; 100 ml, 400 mmol) was added to a suspension of crude tert-6utyl (R) - (1- (3- (2,5-dimethyl-1H-pyrrol-1-yl) -2-methylphenyl ) propan-2-yl) carbamate in MeOH (200 ml) and DCM (50 ml). The resulting red solution was stirred at room temperature for 4 hours and then concentrated under reduced pressure. The resulting brown solid was used in the next step without further purification.

Mass / charge: ES ⁺ [M + H] ⁺ = 243.

Obtaining (R) -3- (2-aminopropyl) -2-methylaniline

A mixture of crude (R) -1- (3- (2,5-dimethyl-1H-pyrrol-1-yl) -2-methylphenyl) propan-2-amine dihydrochloride, an aqueous solution of hydroxylamine (50 wt%; 107 ml , 1.74 mol) and hydroxylamine hydrochloride (97 g, 1.39 mol) in ethanol (400 ml) was heated under reflux conditions. After 18 h, the reaction mixture was cooled to 0 ° C., basified with aqueous sodium hydroxide solution (50 wt%; 153 g, 1.92 mol) and extracted with DCM. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by SFC (DEAP column from Princeton Chromatography, 100 mm long, 30 mm diameter, 5 μm, column temperature 40 ° C, column pressure 100 bar, flow rate 100 mg / ml) eluting with 25% methanol containing 0.2% NH4OH in CO ₂ to give (R) -3- (2aminopropyl) -2-methylaniline (24 g, 84%) as a light amber solid.

1H NMR (500 MHz, DMSO, 27 ° C) 1.05 (3H, d), 1.99 (3H, s), 2.55 (1H, dd), 2.93 (1H, dd), 3, 11- 3.25 (1H, m), 4.77 (2H, s), 6.35 (1H, dd), 6.52 (1H, dd), 6.81 (1H, t).

No NH ₂ protons were observed at alkyl, mass / charge: ES ⁺ [M + H] ⁺ = 165.

Preparation of (R) -N- (1- (3-amino-2-methylphenyl) propan-2-yl) -1-fluorocyclopropane-1-carboxamide

(R) -3- (2-Aminopropyl) -2-methylaniline (1.70 g, 10.4 mmol) was dissolved in DMF (29.9 ml) and treated with 1-fluorocyclopropane-1-carboxylic acid (1, 00 g, 9.61 mmol), HATU (4.02 g, 10.6 mmol) and TEA (2.68 ml, 19.22 mmol). The reaction mixture was stirred at room temperature for 3 h and then quenched with water and extracted with EtOAc. The organic layer was washed with a saturated sodium chloride aqueous solution, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was dried in vacuo overnight to remove residual DMF. The residue was then adsorbed on silica and purified by flash column chromatography on silica, elution gradient from 0 to 80% ethyl acetate in hexanes, to give (R) -N- (1- (3-amino-2-methylphenyl) propane -2yl) -1-fluorocyclopropane-1-carboxamide (1.47 g, 61.1%) as a light yellow solid.

¹ H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.01-1.28 (7H, m), 2.02 (3H, s), 2.54-2.62 (1H, m) , 2.83 (1H, dd), 3.89-4.16 (1H, m), 4.68 (2H, s), 6.38 (1H, d), 6.49 (1H, d), 6.78 (1H, t); 8.14 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ 251.

Preparation of (R) -3- (2 - (((1-fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline

Borane-tetrahydrofuran complex in THF (1 M; 35.2 ml, 35.2 mmol) was added to a solution of (R) -N- (1- (3-amino-2-methylphenyl) propan-2-yl) -1- fluorocyclopropane-1-carboxamide (1.47 g, 5.87 mmol) in THF (13.7 ml) at room temperature under nitrogen atmosphere. The reaction mixture was then heated at 65 ° C for 6 hours. The reaction mixture was cooled to 0 ° C and carefully quenched with MeOH (evolution of gas). The solution was then concentrated under reduced pressure and stored in a freezer

58 037533 over 18 hours. The residue was dissolved in MeOH (6 ml) and heated at 65 ° C for 3 hours. The solution was cooled to room temperature and then concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 16% methanol in DCM, to give (R) -3- (2 - (((1-fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (1.14 g, 82%) as a colorless oil.

1H NMR (300 MHz, DMSO-d6, 27 ° C) 0.54-0.70 (2H, m), 0.79-1.02 (5H, m), 1.64 (1H, br. S. ), 1.99 (3H, s), 2.36 (1H, dd), 2.69-2.97 (4H, m), 4.68 (2H, s), 6.36 (1H, d) , 6.49 (1H, d); 6.79 (1H, t);

mass / charge: ES ⁺ [M + H] ⁺ = 237.

Preparation of (1 S, 3R) -1 - (5-bromopyridin-2-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine

5-Bromopicolinaldehyde (507 mg, 2.72 mmol) was added to a stirred solution of (R) -3- (2 - (((1fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (322 mg, 1.36 mmol) in acetic acid (7028 μl) and water (143 μl). The resulting mixture was heated to 90 ° C and stirred at this temperature for 5 h. The mixture was concentrated and dissolved in EtOAc (50 ml) and saturated NaHCO ₃ (25 ml). The aqueous layer was extracted with EtOAc (2x50 ml). The combined organics were concentrated under reduced pressure. The residue was dissolved in DCM (20 ml) and 1 M HCl (20 ml). The mixture was vigorously stirred for 30 minutes. The layers were separated and the aqueous layer was washed with DCM (20 ml). The aqueous layer was podslushivaet to pH> 10 with solid NaOH. The solution was extracted with DCM (3x25 ml). The combined organics were dried (phase separator cartridge) and concentrated to give the crude product. The crude product was purified by flash chromatography on silica, elution gradient 0 to 60% EtOAc in heptane. Pure fractions were evaporated to dryness to give (1S, 3R) -1- (5-bromopyridin-2-yl) -2 ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4- tetrahydroisoquinoline-6-amine (217 mg, 39%) as a yellow gum.

1H NMR (500 MHz, CDC13, 27 ° C) 0.33-0.47 (1H, m), 0.49-0.6 (1H, m), 0.89-1.01 (2H, m) , 1.04 (3H, d), 2.06 (3H, s), 2.55- 2.64 (2H, m), 2.92-3.04 (2H, m), 3.51 (2H , s), 3.65-3.72 (1H, m), 4.88 (1H, s), 6.45 (1H, d), 6.58 (1H, d), 7.31 (1H, dd), 7.66 (1H, dd), 8.54 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 404.

Obtaining (6S, 8R) -6- (5-bromopyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3- f] isoquinoline

(1 S, 3R) -1 - (5-Bromopyridin-2-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine ( 295 mg, 0.73 mmol) in propionic acid (8291 μl) was cooled to -20 ° C. Sodium nitrite (50.3 mg, 0.73 mmol) in water (829 μl) was added dropwise over 2-3 minutes. The reaction mixture was stirred at -20 ° C for 45 minutes. The reaction mixture was diluted with ice cold EtOAc (30 ml). The reaction mixture was poured into saturated NaHCO ₃ (40 ml). The mixture was stirred vigorously for 5 minutes. The layers were separated and the organic layer was washed with saturated NaHCO ₃ (2x30 ml). The combined aqueous layers were extracted again with EtOAc (2x30 ml). The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated to give the crude product as a brown oil. The crude material was purified by flash column chromatography on silica eluting with 0-45% EtOAc in heptane to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4.3- £] isoquinoline (107 mg, 35%) as an orange solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.35-0.44 (1H, m), 0.51-0.63 (1H, m), 0.92-1.07 (2H, m ), 1.09 (2H, d), 2.69 (1H, dd), 2.96 (1H, dd), 3.01-3.10 (1H, m), 3.33-3.49 ( 2H, m), 3.78-3.89 (1H, m), 5.03 (1H, s), 6.95 (1H, d), 7.19 (1H, d), 7.34-7 , 39 (1H, m), 7.68 (1H, dd), 8.07 (1H, d), 8.58 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 415 and 417.

Obtaining (6S, 8R) -6- (5-bromopyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro2H-pyran-2-yl) -6.7, 8,9-tetrahydro-3H-pyrazolo [4,3] isoquinoline

- 59 037533

3,4-Dihydro-2H-pyran (0.029 ml, 0.32 mmol) was added to a solution of (6S, 8R) -6- (5-brompyridin-2yl) -7 - ((1-fluorocyclopropyl) methyl) -8- methyl 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (107 mg, 0.26 mmol) and 4-methylbenzenesulfonic acid hydrate (9.80 mg, 0.05 mmol) in DCM (2 ml) and the reaction mixture was heated to 45 ° C for 3 h. Additional 3,4-dihydro-2Hpyran (0.235 ml, 2.58 mmol) and 4-methylbenzenesulfonic acid hydrate (49.0 mg, 0 , 26 mmol) and the mixture was heated at 45 ° C for 16 h. The reaction mixture was diluted with DCM and washed with saturated NaHCO ₃ solution. The organic phase was dried and evaporated, then the crude material was passed through a plug of silica (EtOAc / heptane 1: 1). The filtrate was evaporated to give (6S, 8R) -6- (5-brompyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline as a brown oil, which was used without further purification.

Mass / charge: ES + [M + H] ⁺ = 499.

Example 4.

N- (4 - ((6S, 8R) -7- (2-Fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,31] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine (diastereoisomeric mixture)

1- (3-Fluoropropyl) azetidin-3-amine (50 mg, 0.37 mmol), mixture (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 (2-fluoro-3-methoxy -2-methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline and (6S, 8R) - 6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) 8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8 , 9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline (140 mg, 0.25 mmol), cesium carbonate (163 mg, 0.50 mmol) and [(2-dicyclohexylphosphino-3,6-dimethoxy- 2 ', 4', 6'triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'-biphenyl)] palladium (P) methanesulfonate (BrettPhos Pd G3) (23 mg, 0, 02 mmol) was suspended in 1,4-dioxane (2 ml) and sealed in a microwave tube. The reaction mixture was heated to 100 ° C for 4 hours under the influence of microwave radiation. The reaction mixture was diluted with DCM (25 ml) and washed with water (25 ml). The organic layer was evaporated. The residue was dissolved in DCM (3 ml) and TFA (1 ml). The reaction mixture was stirred at room temperature for 1 hour and then partitioned between DCM and 2M NaOH (20 ml each). The organic phase was evaporated and the crude product was purified by preparative HPLC (Waters CSH C18 OBD column, 5 μm silica, 30 mm in diameter, 100 mm in length) using mixtures of water (containing 1% NH ₃ ) and MeCN with decreasing polarity as eluents to obtain N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9 tetrahydro-3H-pyrazolo [4,3 -f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine (26.0 mg, 20%) as a resin, which is a mixture of diastereoisomers.

1H NMR (500 MHz, DMSO-d6, 27 ° С) 0.94-0.99 (3H, m), 1.16-1.24 (3H, m), 1.64 (2H, dq), 2 , 39-2.47 (3H, m), 2.70 (2H, t), 2.74-2.84 (2H, m), 3.03- 3.21 (2H, m), 3.24 (3H, d), 3.48 (2H, dd), 3.58-3.64 (2H, m), 3.79 (3H, d), 3.88 -3.94 (1H, m), 4.44 (2H, dt), 5.17 (1H, d), 5.88 (1H, td), 5.97 (1H, dd), 6.16 (1H, d), 6.32-6 , 38 (1H, m), 6.63 (1H, t), 7.16 (1H, dd), 8.02 1H, s), 12.91 (1H, s);

¹⁹ F NMR (471 MHz, DMSO-d ₆ , 27 ° C) -218.19, -149.21, -148.22;

mass / charge: ES ⁺ [M + H] ⁺ = 528.

A mixture of (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl3- (tetrahydro-2H-pyran-2-yl ) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7- (2-fluoro-3-methoxy -2-methylpropyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9 tetrahydro-2H-pyrazolo [4,3-f] isoquinoline as follows.

Preparation of a diastereoisomeric mixture of N - ((R) -1- (3-bromo-2-methylphenyl) propan-2-yl) -2-fluoro-3methoxy-2-methylpropan-1-amine

- 60 037533

Trifluoromethanesulfonic acid anhydride (2.148 ml, 12.79 mmol) was added to ((2-fluoro-3methoxy-2-methylpropoxy) methanetriyl) tribenzene (4.44 g, 12.18 mmol) dissolved in dichloromethane (50 ml) and the reaction mixture was stirred at room temperature for 30 minutes. Then triethylsilane (2.140 ml, 13.40 mmol) was added and the mixture was stirred for an additional 30 minutes. The reaction mixture was concentrated to give the crude racemic 2-fluoro-3-methoxy-2methylpropyl trifluoromethanesulfonate, which was used directly in the next step without further purification.

DIPEA (5.36 ml, 30.68 mmol) was added to (R) -1- (3-bromo-2-methylphenyl) propan-2-amine (2.80 g, 12.3 mmol) and 2-fluoro- 3-methoxy-2-methylpropyltrifluoromethanesulfonate (3.12 g, 12.3 mmol) in 1,4-dioxane (60 ml) at room temperature under nitrogen [exothermic]. The resulting mixture was stirred at 85 ° C for 24 hours. The reaction mixture was allowed to cool and the reaction mixture was evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 60% EtOAc in heptane to give N - ((R) -1- (3-6pom-2methylphenyl) propan-2-yl) -2- fluoro-3-methoxy-2-methylpropan-1-amine (2.480 g, 73%) as an oil, which is a mixture of diastereoisomers.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.02-1.07 (3H, m), 1.31 (1.5H, d), 1.32 (1.5H, d), 2 , 41 (3H, s), 2.572.66 (1H, m), 2.68-2.90 (4H, m), 3.34 (1.5H, d), 3.35 (1.5H, d ), 3.38-3.46 (2H, m), 6.96 (1H, t), 7.07 (1H, d), 7.42 (1H, d);

NH not observed ¹⁹ F NMR (471 MHz, CDCl ₃ , 27 ° C) -157.94, -157.53;

mass / charge: ES ⁺ [M + H] ⁺ = 332/334.

Preparation of a diastereoisomeric mixture of 3 - ((R) -2 - ((2-fluoro-3-methoxy-2-methylpropyl) amino) propyl) 2-methylaniline

tris- (Dibenzylideneacetone) dialladium (0) (0.205 g, 0.22 mmol) and (±) -2,2'-bis- (diphenylphosphino) 1,1'-binaphthalene (0.279 g, 0.45 mmol) were added to suspensions of a diastereoisomeric mixture of N - ((R) -1- (3-bromo-2-methylphenyl) propan-2-yl) -2-fluoro-3-methoxy-2-methylpropan-1-amine (2.48 g, 7.46 mmol), benzophenone imine (1.487 g, 8.21 mmol) and sodium tert-butoxide (1.076 g, 11.20 mmol) in degassed toluene (30 ml) and the reaction mixture was heated to 90 ° C for 3 h. After cooling, toluene evaporated. The residue was dissolved in DCM (250 ml) and washed with water (250 ml). The aqueous layer was extracted with DCM (100 ml) and the combined organics were concentrated to approximately 50 ml. 2M HCl solution (50 ml) was added and the biphasic mixture was stirred vigorously for 30 min. The layers were separated and the aqueous phase was extracted with DCM. The aqueous phase was podslushivaet with 2 M aqueous NaOH solution. The resulting was extracted with DCM (2 x 250 ml) and the combined extracts in DCM were evaporated to give 3 - ((R) -2 - ((2-fluoro-3-methoxy-2methylpropyl) amino) propyl) -2-methylaniline (2 , 00 g, 100%) in the form of an oil, which is a mixture of diastereoisomers.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.04 (3H, d), 1.31 (1.5H, d), 1.32 (1.5H, d), 2.11 (3H, s), 2.53-2.60 (1H, m), 2.70-2.89 (4H, m), 3.35 (2H, d), 3.36 (1.5H, d), 3 , 43 (1.5H, dd), 3.58 (2H, s), 6.57 (1H, d), 6.61 (1H, d), 6.94 (1H, t), 7.58 ( 1H, s);

¹⁹ F NMR (471 MHz, CDCl ₃ , 27 ° C) -157.61, -157.30.

mass / charge: ES ⁺ [M + H] ⁺ = 269.

Preparation of a diastereoisomeric mixture (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2-fluoro-3-methoxy-2methylpropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline -6-amine

Bg

Diastereoisomeric mixture of 3 - ((2R) -2 - ((2-fluoro-3-methoxy-2-methylpropyl) amino) propyl) -2methylaniline (2.00 g, 7.45 mmol) and 4-bromo-2-methoxybenzaldehyde (3.37 g, 15.6 mmol) was heated in acetic acid (30 ml) and water (0.671 ml, 37.3 mmol) to 70 ° C overnight. After cooling, the acetic acid was evaporated. The residue was dissolved in EtOAc (40 ml) and 2N. HCl solution (40 ml). The biphasic mixture was stirred for 30 min, then the layers were separated. The organic phase was extracted with water, then the combined aqueous phases were podslushivaet by the addition of 2N. NaOH solution and extracted with DCM (2x200 ml). The combined DCM layers were evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane to first give (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2-fluoro- 61 037533

3-methoxy-2-methylpropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (0.354 g, 10%) as one diastereoisomer.

¹ H NMR (500 MHz, DMSO-d ₆ , 27 ° С) 0.91 (3H, d), 1.22 (3H, d), 1.94 (3H, s), 2.26-2.40 (2H, m), 2.65-2.78 (2H, m), 3.16-3.25 (4H, m), 3.42 (1H, dd), 3.85 (3H, s), 4.61 (2H, s), 5.10 (1H, s), 6.24 (1H, d), 6.37 (1H, d),

6.61 (1H, d), 6.95 (1H, dd), 7.15 (1H, d).

One proton is partially blocked by water.

¹⁹ F NMR (471 MHz, DMSO-d _f "27 ° C) -149.65;

mass / charge: ES + [M + H] ⁺ = 465/467.

Then, fractions were obtained containing a mixture of diastereoisomers (1S, 3R) -1- (4-6pom-2methoxyphenyl) -2- (2-fluoro-3-methoxy-2-methylpropyl) -3,5-dimethyl-1,2,3 , 4-tetrahydroisoquinoline-6-amine (1.700 g, 49%).

¹⁹ F NMR (471 MHz, DMSO-d6, 27 ° C) -149.65, -148.78;

mass / charge: ES ⁺ [M + H] ⁺ = 465/467.

Then, fractions were obtained containing the second diastereoisomer (1S, 3R) -1- (4-bromo-2-methoxyphenyl) 2- (2-fluoro-3-methoxy-2-methylpropyl) -3,5-dimethyl-1,2, 3,4-tetrahydroisoquinoline-6-amine (0.274 g, 8%) as one diastereoisomer.

¹ H NMR (500 MHz, DMSO-d6, 27 ° C) 0.91 (3H, d), 1.16 (3H, d), 1.93 (3H, s), 2.26-2.4 ( 2H, m), 2.63-2.74 (2H, m), 3.23 (3H, s), 3.35 (1H, dd), 3.47 (1H, dd), 3.85 (3H , s), 4.60 (2H, s), 5.09 (1H, s), 6.24 (1H, d), 6.37 (1H, d), 6.61 (1H, d), 6 , 93 (1H, dd), 7.15 (1H, d).

One proton is blocked by water.

¹⁹ F NMR (471 MHz, DMSO-d6, 27 ° C) -148.78;

mass / charge: ES ⁺ [M + H] ⁺ = 465/467.

Everything is in the form of resins.

Preparation of a diastereoisomeric mixture (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H -pyrazolo [4,3-f] isoquinoline

A solution of sodium nitrite (0.277 g, 4.02 mmol) in water (1.0 ml) was added to a solution of the diastereoisomeric mixture (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2-fluoro- 3-methoxy-2-methylpropyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinolin-6-amine (1.70 g, 3.65 mmol) in propionic acid (15 ml) at -10 ° C. The reaction mixture was stirred for 1 h, then ice-cold EtOAc (20 ml) was added. The reaction mixture was quenched by the addition of an aqueous solution of NaHCO ₃ (30 ml) and stirred for 15 min before being allowed to warm to room temperature. The organic phase was washed with aqueous NaHCO ₃ (30 ml) and brine (20 ml), dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2fluoro-3- methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.950 g, 55%) as a brown solid, a mixture of diastereoisomers.

L NMR (500 MHz, DMSO-d6, 27 ° C) 0.95-0.97 (3H, m), 1.14-1.23 (3H, m), 2.31-2.40 (1H, m), 2.78-2.88 (2H, m), 3.11-3.21 (2.5H, s), 3.23 (1.5H, s), 3.32-3.38 ( 1H, m), 3.40-3.52 (2H, m), 3.88 (1.5H, s), 3.89 (1.5H, s), 5.28 (1H, s), 6 , 59 -6.69 (2H, m), 6.92-6.96 (1H, m), 7.16-7.22 (2H, m), 8.05 (1H, s), 12.96 (1H, s);

¹⁹ F NMR (471 MHz, DMSO-d _f "27 ° C) -149.65, -148.68;

mass / charge: ES ⁺ [M + H] ⁺ = 476/478.

Preparation of a mixture (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8methyl-3- (tetrahydro-2H-pyran-2-yl ) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (4bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy -2-methylpropyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) 6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline

4-methylbenzenesulfonic acid hydrate (0.038 g, 0.20 mmol) was added to a solution of the diastereoisomeric mixture (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2- methylpropyl) -8-methyl

- 62 037533

6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.950 g, 1.99 mmol) and 3,4-dihydro-2H-pyran (0.546 ml, 5.98 mmol) in DCM (30 ml) and the mixture was heated at 40 ° C overnight. Additional 3,4-dihydro-2H-pyran (0.546 ml, 5.98 mmol) and 4-methylbenzenesulfonic acid hydrate (0.038 g, 0.20 mmol) were added. Heating was continued for 7 hours. Additional portions of 3,4-dihydro-2H-pyran (0.546 ml, 5.98 mmol) and 4-methylbenzenesulfonic acid hydrate (0.038 g, 0.20 mmol) were added and the reaction mixture was heated overnight ... The reaction mixture was diluted with DCM (50 ml) and washed with saturated aqueous NaHCO ₃ solution (50 ml).

The organic phase was evaporated to a dark brown oil and the crude product was purified by flash chromatography on silica, elution gradient 0 to 35% EtOAc in heptane, to give (6S, 8R) -6- (4-bromo-2 -methoxyphenyl) -7- (2-fluoro-3-methoxy-2methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3-f] isoquinoline and (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-2- (tetrahydro -2Hpyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline (0.960 g, 86%) as a foam, which is a mixture of diastereoisomers.

¹⁹ F NMR (471 MHz, DMSO-d6, 27 ° C) -149.79, -149.74, -149.69, -149.65, -148.77, -148.73, -148.69, 148.66;

mass / charge: ES ⁺ [M + H] ⁺ = 560/562.

Example 5.

3 - ((6S, 8R) -6- (2,6-Difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8, 9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropan-1-ol

TBAF in THF (1 M; 160 μL, 0.16 mmol) was added to a solution of N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8 -methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3-fluoropropyl) azetidin-3-amine ( 80 mg, 0.10 mmol) in THF (0.5 ml) at room temperature. After 4 h, the reaction mixture was concentrated under reduced pressure and the resulting residue was purified by flash chromatography on silica, elution gradient 30% to 90% (10% MeOH in DCM containing 1% NH4OH) in DCM. The product fractions were concentrated under reduced pressure and the resulting residue was purified again using similar conditions. The product fractions were again concentrated under reduced pressure and the resulting residue was purified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5 μm silica, 19 mm in diameter, 100 mm in length) using water mixtures (containing 0.2% ammonium hydroxide ) and MeCN with decreasing polarity as eluents (from 40 to 70% over 7 min.) to give 3 - ((6S, 8R) -6- (2,6-difluoro-4 - ((1- (3- fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropane- 1-ol (17 mg, 31%) as a white solid.

Ή NMR (500 MHz, DMSO-d6, 27 ° С) 1.02 (3H, d), 1.59-1.69 (2H, m), 2.44 (2H, t), 2.56-2 , 67 (1H, m), 2.72 (2H, t), 2.88 (1H, dd), 3.02-3.13 (1H, m), 3.17 (1H, dd), 3, 49-3.56 (1H, m), 3.57-3.69 (3H, m), 3.91 (1H, tdt), 4.44 (2H, dt), 5.08 (1H, s) , 5.25 (1H, br. S), 6.07 (2H, d), 6.64 (1H, d), 6.69 (1H, d), 7.20 (1H, d), 8, 04 (1H, s), 12.95 (1H, br. S).

H alone was not observed, mass / charge: ES ⁺ [M + H] ⁺ = 524.

N- (4 - ((6S, 8R) -7- (3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro3H-pyrazolo [4 , 3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1 - (3-fluoropropyl) azetidine-3-amine was prepared as follows.

Preparation of 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-ol

NaH in mineral oil (60 wt%; 343 mg, 8.58 mmol) was added in one portion to a stirred solution of 2,2-difluoropropane-1,3-diol (874 mg, 7.80 mmol) in THF (32 ml) at 0 ° C. Provided

- 63 037533 heating the reaction mixture to room temperature and stirring at room temperature for 2 hours. The reaction mixture was again heated to 0 ° C. and tert-butyldiphenylchlorosilane (2.0 ml, 7.8 mmol) was added dropwise via a syringe. The reaction mixture was allowed to warm to room temperature over 1 h and then quenched with water and extracted with EtOAc. The organic layer was dried with Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with isocratic 5% ethyl acetate in hexanes to give 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-ol (1.94, 71%) as colorless oil.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° С) δ ppm 1.03-1.14 (9H, s), 3.87-3.93 (4H, m), 7.37-7.44 (6H, m), 7.64-7.66 (4H, m).

Preparation of 3 - ((tert-butyldiphenylsilyl) oxy) -2., 2-difluoropropyltrifluoromethanesulfonate

A solution of 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-ol (1.94 g, 5.55 mmol) and 2,6-dimethylpyridine (1.94 ml, 16.6 mmol) in DCM (18 ml) was cooled to -10 ° C (salt / ice bath). Trifluoromethanesulfonic anhydride (1.88 mL, 11.1 mmol) was slowly added dropwise over 10 minutes. The reaction mixture was maintained under these conditions for 2 hours. The reaction mixture was then washed with water, aqueous HCl (1N; 100 ml) and saturated aqueous sodium bicarbonate solution. The organic layer was dried over MgSO ₄ , filtered and concentrated under reduced pressure to give 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl trifluoromethanesulfonate (2.68 g, 100%) as a red oil.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° С) 1.03-1.14 (9H, s), 3.90 (2H, t), 4.76 (2H, t), 7.39- 7.56 (6H, m), 7.59-7.75 (4H, m).

Preparation of 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyltrifluoromethanesulfonate

3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyltrifluoromethanesulfonate (1.92 g, 3.98 mmol) was added to a solution of (R) -3- (2-aminopropyl) -2-methylaniline (0.784 g, 4 , 77 mmol) and DIPEA (1.031 ml, 5.97 mmol) in 1,4-dioxane (15 ml). The reaction mixture was heated at 85 ° C for 18 h. After cooling, the reaction mixture was diluted with DCM and washed with water. The aqueous layer was extracted with DCM and the combined organic layers were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, gradient elution from 0 to 4% methanolic ammonia in DCM. Pure fractions were concentrated to dryness to obtain (R) -3- (2 - ((3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) amino) propyl) -2-methylaniline (1.97 g, 100% ) as a yellow oil.

¹ H NMR (300 MHz, METHANOL ^, 27 ° C) δ ppm 0.97-1.12 (12H, m), 2.10 (3H, s), 2.53-2.63 (1H, m) , 2.74-2.84 (1H, m), 2.86-2.99 (1H, m), 3.00-3.19 (2H, m), 3.80 (2H, td), 6 , 53 (1H, d), 6.63 (1H, d), 6.86 (1H, t), 7.38-7.50 (6H, m), 7.64-7.72 (4H, m );

mass / charge: ES ⁺ [M + H] ⁺ = 497.

Obtaining (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2difluoropropyl) -3,5-dimethyl-1,2, 3,4-tetrahydroisoquinoline-6-amine

4-Bromo-2,6-difluorobenzaldehyde (1.55 g, 7.02 mmol) was added to a solution of (R) -3- (2 - ((3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) amino ) propyl) -2-methylaniline (1.74 g, 3.51 mmol) in acetic acid (17 ml) and water (0.32 ml, 18 mmol) and the reaction mixture was heated at 80 ° C overnight. The reaction mixture was concentrated under reduced pressure and the residue was partitioned between

- 64 037533

EtOAc and saturated aqueous NaHCO ₃ solution. The organic layer was concentrated under reduced pressure and purified by flash silica chromatography, gradient elution from 20 to 80% ethyl acetate in hexanes to give (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2 - (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (1.33 g, 54.5%) as solid matter.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° С) δ ppm 1.04-1.11 (12H, m), 2.25-2.38 (3H, m), 2.53-2.65 (1H, m), 2.73 (1H, q), 2.86-3.03 (1H, m), 3.15-3.38 (1H, m), 3.52-3.71 (2H , m), 3.85-4.01 (1H, m), 5.31 (1H, d), 6.58-6.64 (1H, m), 6.67-6.73 (1H, m ), 6.88-6.95 (2H, m), 7.18-7.24 (2H, m), 7.37-7.50 (6H, m), 7.60-7.70 (4H , m);

mass / charge: ES ⁺ [M + H] ⁺ = 699.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2.2difluoropropyl) -8-methyl-6,7,8, 9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinoline

Br

Sodium nitrite (0.130 g, 1.89 mmol) in water (0.900 ml) was added dropwise to a solution of (1S, 3R) -1- (4brom-2,6-difluorophenyl) -2- (3 - ((tert-butyldiphenylsilyl ) oxy) -2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine (1.32 g, 1.89 mmol) in propionic acid (9 ml) at -8 ° C (salt / ice bath) and stirred under these conditions for 20 minutes. The reaction mixture was diluted with ethyl acetate (20 ml, pre-cooled to -10 ° C) and quenched by the slow addition of saturated aqueous NaHCO ₃ solution (30 ml, pre-cooled to -10 ° C) over 15 min at 0 ° C. The mixture was allowed to warm to room temperature and maintained under such conditions for 18 h. The organic layer was separated and the aqueous layer was extracted with EtOAc. The combined organic layers were washed with saturated aqueous NaHCO ₃ and water, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 15 to 50% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 (( tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrαzolo [4,3f] isoquinoline (0.520 g, 38.8%) as light brown solid.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° C) δ ppm 1.07 (9H, s), 1.12 (3H, d), 2.72-2.92 (2H, m), 3.213, 39 (2H, m), 3.51-3.66 (1H, m), 3.67-3.79 (1H, m), 3.86-4.00 (1H, m), 5.37 ( 1H, s), 6.81 (1H, d), 6.87-6.98 (2H, m), 7.25-7.29 (1H, m), 7.35-7.50 (6H, m), 7.59-7.70 (4H, m), 8.14 (1H, s).

NH indole was not observed, mass / charge: ES ⁺ [M + H] ⁺ = 710.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2difluoropropyl) -8-methyl-3- (tetrahydro-2H -pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrαzolo [4,3f] isoquinoline

Br

3,4-Dihydro-2H-pyran (95 μL, 1.04 mmol) was added to a solution of (6S, 8R) -6- (4-6pom-2,6 difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) hydroxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrαzolo [4,3-f] isoquinoline (492 mg, 0.69 mmol) and para-toluenesulfonic acid monohydrate (13 mg, 0.07 mmol) in DCM (3.5 ml) and the reaction mixture was heated under reflux for 6 h. After cooling, the reaction mixture was washed with saturated aqueous NaHCO3 solution, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure ... The residue was purified by flash chromatography on silica eluting with 10 to 30% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - (( tert-butyldiphenylsilyl) oxy) 2,2-difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (474 mg, 86%) as a light brown gummy solid.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° С) 1.06 (9H, s), 1.19-1.25 (3H, m), 1.74-1.84 (3H, m), 2.092.20 (2H, m), 2.49-2.67 (1H, m), 2.84-2.96 (2H, m), 3.27-3.45 (2H, m), 3, 61-3.87 (3H, m), 4.04 (2H, d), 5.52 (1H, s), 5.66-5.72 (1H, m), 6.78 (1H, d) , 6.94 (2H, d), 7.32 (1H, d), 7.36-7.50 (6H, m), 7.60-7.68 (4H, m), 8.04

- 65 037533 (1H, s).

Preparation of tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8methyl-3- (tetrahydro-2H-nyran- 2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5 difluorophenyl) amino) azetidine-1-carboxylate

Degassed mixture (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) 2,2-difluoropropyl) -8-methyl-3- (tetrahydro -2H-nyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (0.474 g, 0.600 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (0 , 15 g, 0.89 mmol), Xantphos (0.069 g, 0.12 mmol), Pd ₂ (dba) ₃ (0.06 g, 0.06 mmol) and cesium carbonate (0.389 g, 1.19 mmol) in dioxane (3 ml) was heated at 115 ° C for 4 h. Then the reaction mixture was concentrated under reduced pressure and the resulting residue was adsorbed on silica gel and purified by flash chromatography on silica, elution gradient from 5 to 50% ethyl acetate in hexanes, with obtaining tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-3 (tetrahydro-2H-nyran -2-yl) -6,7,8,9-tetrahydro-3H-n-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5 difluorophenyl) amino) azetidine-1-carboxylate (0.377 g, 71, 3%) as a solid.

¹ H NMR (300 MHz, CHLOROFORM-d, 27 ° С) 1.06 (9H, s), 1.12 (3H, d), 1.46 (9H, s), 1.65-1.84 ( 3H, m), 2.07-2.23 (2H, m), 2.58 (1H, d), 2.75-2.91 (2H, m), 3.16-3.34 (2H, m), 3.52-3.78 (5H, m), 3.91-4.07 (3H, m), 4.19-4.29 (2H, m), 5.23-5.34 ( 1H, m), 5.67 (1H, dd), 5.81 (2H, d), 6.81 (1H, d), 7.25-7.29 (1H, m), 7.35-7 , 48 (6H, m), 7.65 (4H, ddd), 8.02 (1H, s).

NH aniline was not observed, weight / charge: ES ⁺ [M + H] ⁺ = 886.

Obtaining N- (4 - ((6S, 8R) -7- (3- (tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4 , 3-f] isoquinolin-6-yl) -3,5-difluorophenyl) azetidin-3-amine

TFA (328 μl, 4.25 mmol) was slowly added to a solution of tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) - 8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) amino ) azetidine-1-carboxylate (377 mg, 0.43 mmol) in DCM (2 ml) at 0 ° C. The reaction mixture was allowed to warm to room temperature and stirred under such conditions for 1 hour. The reaction mixture was then concentrated under reduced pressure and the resulting residue was partitioned between ethyl acetate and saturated aqueous NaHCO ₃ solution. The organic layer was dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 30 to 70% (10% methanol in DCM containing 1% ammonium hydroxide) in DCM to give N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3, 5-difluorophenyl) azetidin-3-amine (166 mg, 55.6%) as a pale solid.

1H NMR (300 MHz, CHLOROFORM-d, 27 ° С) 0.96-1.20 (12H, m), 2.62-3.03 (2H, m), 3.10-3.42 (2H, m), 3.42-3.63 (3H, m), 3.63-3.80 (2H, m), 3.81-4.08 (3H, m), 4.15-4.28 ( 1H, m), 4.38 (1H, d), 5.23 (1H, s), 5.83 (2H, m), 6.80 (1H, m), 7.15 (1H, d), 7.32-7.53 (6H, m), 7.61-7.81 (4H, m), 8.06 (1H, s).

NH indazole was not observed, mass / charge: ES ⁺ [M + H] ⁺ = 702.

Obtaining N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [ 4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3-fluoropropyl) azetidin-3-amine

- 66 037533

1-Fluoro-3-iodopropane (40 mg, 0.21 mmol) was added to a solution of N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) - 8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) azetidin-3-amine (144 mg, 0.21 mmol ) and DIPEA (107 μl, 0.62 mmol) in DMF (1 ml) at ambient temperature. The reaction mixture was stirred at room temperature for 3 h and then diluted with EtOAc (40 ml) and washed with saturated aqueous sodium chloride solution (3x20 ml). The organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient 15% to 30% (10% methanol in DCM containing 1% ammonium hydroxide) in DCM to give N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) - 3,5-difluorophenyl) -1- (3-fluoropropyl) azetidin-3-amine (87 mg, 56%) as a yellow solid.

Mass / charge: ES + [M + H] ⁺ = 762.

Example 6.

N- (4 - ((6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,31] isoquinolin-6-yl ) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine F

Q

NH n /] N --- H F kJ '- /

1-Fluoro-3-iodine propane (40.9 μL, 0.39 mmol) was added to a solution of N- (4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3methoxyphenyl) azetidine-3-amine (169 mg, 0.39 mmol) and DIPEA (203 μL, 1.16 mmol ) in NMP (1.7 ml) at room temperature. After 18 hours, the reaction mixture was concentrated under reduced pressure. The resulting residue was purified by reverse phase (C 18) flash chromatography eluting with mixtures of water (containing 0.2% ammonium hydroxide) and decreasing polarity MeCN as eluants. The product fractions were combined and lyophilized to give N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4 , 3-f] isoquinolin6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine (75 mg, 39%) as a clear residue.

1H NMR (500 MHz, DMSO-d6, 27 ° C) 0.39-0.46 (1H, m), 0.47-0.54 (1H, m), 0.77-0.85 (2H, m), 0.91 (3H, d), 1.51-1.64 (2H, m), 2.38 (2H, t), 2.54 (1H, dd), 2.63 (2H, q ), 2.75-2.90 (2H, m), 3.14 (1H, dd), 3.52-3.64 (3H, m), 3.73 (3H, s), 3.79- 3.88 (1H, m), 4.38 (2H, dt), 5.10 (1H, s), 5.82-5.87 (1H, m), 5.91 (1H, d), 6 , 09 (1H, d), 6.49 (1H, d), 6.57 (1H, d), 7.07 (1H, d), 7.94 (1H, s), 12.83 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 496.

N- (4 - ((6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,31] isoquinolin-6-yl ) -3-methoxyphenyl) azetidin-3-amine was prepared as follows.

Preparation of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine

- 67 037533

(R) -3- (2 - (((1-Fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (451 mg, 1.91 mmol) and 4-bromo-2-methoxybenzaldehyde (410 mg, 1.91 mmol) in a mixture of water (172 mg, 9.54 mmol) and acetic acid (7.5 ml) was heated at 80 ° C for 6 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was treated with an aqueous solution of HCl (1N .; 18 ml). The mixture was stirred at 80 ° C for 18 hours. Then the reaction mixture was cooled and saturated aqueous NaHCO ₃ solution was added. The mixture was extracted with DCM and the organic layer was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 5% to 35% ethyl acetate in hexanes, to give (1S, 3R) -1- (4-6pom2-methoxyphenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (591 mg, 71.5%) as a resinous film.

¹ H NMR (300 MHz, DMSO-de, 27 ° C) 0.42-0.61 (2H, m), 0.81-0.91 (2H, m), 0.93 (3H, d), 1.93 (3H, s), 2.43-2.48 (2H, m), 2.72-2.89 (2H, m), 3.48-3.57 (1H, m), 3, 87 (3H, s), 4.57 (2H, s), 5.12 (1H, s), 6.24 (1H, d), 6.35 (1H, d), 6.82 (1H, d ), 6.96 (1H, d), 7.16 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 433.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3- f] isoquinoline

Acetic acid (0.392 ml, 6.84 mmol) was added to (1s, 3R) -1- (4-bromo-2-methoxyphenyl) -2 - ((1fluorocyclopropyl) methyl) -3,5-dimethyl-1,2, 3,4-tetrahydroisoquinoline-6-amine (0.593 g, 1.37 mmol) in CHCl ₃ (5 ml) and the resulting solution was cooled to 0 ° C. Isopentyl nitrite (0.321 g, 2.74 mmol) in CHCl ₃ (1 ml) was added dropwise to the reaction mixture and the reaction mixture was stirred at 0 ° C for an additional 2 h. Then a saturated aqueous solution of NaHCO ₃ (1 g in 20 ml of water). The reaction mixture was stirred at 0 ° C for 10 min and then the layers were separated. The organic layer was purified by flash column chromatography eluted from 5% to 35% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) - 8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.299 g, 49.2%) as a light brown solid.

Ή NMR (300 MHz, DMSO-d6, 27 ° C) δ ppm 0.39-0.65 (2H, m), 0.77-0.96 (2H, m), 1.00 (3H, d) , 2.52-2.63 (1H, m), 2.87-3.03 (2H, m), 3.21-3.28 (1H, m), 3.71 (1H, d), 3 , 92 (3H, s), 5.31 (1H, s), 6.65 (1H, d), 6.86-6.92 (1H, m), 6.97 (1H, m), 7, 18 (1H, d), 7.23 (1H, d), 8.05 (1H, s), 12.94 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 444.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline Br m3

I ° \

X .— /

3,4-Dihydro-2H-pyran (176 μL, 1.93 mmol) was added to a solution of (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8- methyl 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (286 mg, 0.64 mmol) and p-toluenesulfonic acid monohydrate (12 mg, 0.060 mmol) in DCM (2, 5 ml). The reaction mixture was exposed to microwave conditions (100 ° C, 300 watts) for 6 hours. On cooling, the reaction mixture was washed with saturated aqueous NaHCO ₃ solution, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 10 to 30% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3- 68 037533

1] isoquinoline (299 mg, 88%) as a light brown gummy solid.

Mass / charge: ES + [M + H] ⁺ = 528.

Preparation of tert-butyl-3 - ((4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine-1carboxylate <7

NH

I ° d = \ \ ^{4 N} - \ / ^F (6S, 8R) -6- (4-Bromo-2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro- 2Нpyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (278 mg, 0.53 mmol), tert-butyl 3aminoazetidine-1-carboxylate (136 mg , 0.79 mmol), cesium carbonate (343 mg, 1.05 mmol) and BrettPhos 3rd generation precatalyst (24 mg, 0.030 mmol) in dioxane (2.5 ml) were exposed to microwave irradiation (100 ° C, 300 W) for 6 h. Then the reaction mixture was concentrated under reduced pressure and the resulting residue was adsorbed on silica gel before purification by flash chromatography on silica, elution gradient from 20 to 60% ethyl acetate in hexanes to give tert-butyl-3 - ((4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8,9-tetrahydro-3H -pyrαzolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine-1carboxylate (296 mg, 91%);

mass / charge: ES ⁺ [M + H] ⁺ = 620.

Preparation of N- (4- (6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3 -f] isoquinolin-6-yl ) -3-methoxyphenyl) azetidine-3-amine n

NH \ / r \

Ν ^ /> - /

HCl dioxane (4 M; 1.2 ml, 4.78 mmol) was added dropwise to a solution of tert-butyl-3 - ((4 - ((6S, 8R) -7 ((1-fluorocyclopropyl) methyl) -8- methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4.3 £ | isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine- 1-carboxylate (296 mg, 0.48 mmol) in MeOH (3.5 ml). After stirring at room temperature overnight, the reaction mixture was concentrated under reduced pressure and the resulting residue was dissolved in MeOH (2 ml). This solution was then loaded onto an SCX-2 ion exchange cartridge that had been pretreated with methanol. The cartridge was washed with methanol and then 7N. ammonia in methanol. The product fractions were concentrated under reduced pressure to give N- (4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4, 3- £ | isoquinolin-6-yl) -3-methoxyphenyl) azetidin-3-amine (181 mg, 87%) as a pale solid.

Mass / charge: ES ⁺ [M + H] ⁺ = 436.

Example 7 (6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl -6,7,8,9-tetrahydro-3H-pyrazolo [4,3- £ | isoquinoline

- 69 037533

1-Fluoro-3-iodopropane (5.79 μL, 0.05 mmol) was added to a solution of trifluoroacetic acid salt and (6S, 8R) -6- (4- (azetidin-3-yloxy) -2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.030 g, 0.050 mmol) and DIPEA (0.029 ml, 0.16 mmol ) in NMP (0.52 ml) at room temperature. After 3 h, the reaction mixture was diluted with EtOAc (40 ml) and washed with saturated aqueous sodium chloride solution (3x20 ml). The organic layer was dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient 15 to 30% (MeOH in DCM containing 1% ammonium hydroxide) in DCM to give (6S, 8R) -7 ((1-fluorocyclopropyl) methyl) - 6- (4 - ((1 - (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (5 mg, 18%) as a white solid.

¹ H NMR (500 MHz, Methanol-d4, 26 ° C) 0.45-0.56 (2H, m), 0.85-1.03 (2H, m), 1.12 (3H, d), 1.67-1.81 (2H, m), 2.50-2.62 (1H, m), 2.61-2.68 (2H, m), 2.92 (1H, dd), 3, 08-3.21 (3H, m), 3.25-3.28 (1H, m), 3.70-3.84 (3H, m), 3.86-3.91 (3H, m), 4.45 (2H, dt), 4.73-4.82 (1H, m), 5.49 (1H, br. S), 6.19 (1H, d), 6.50 (1H, br. s), 6.72 (1H, d), 6.79 (1H, d), 7.18 (1H, d), 8.05 (1H, s).

NH indazole was not observed, mass / charge: ES ⁺ [M + H] ⁺ = 497.

Trifluoroacetic acid salt and N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline -6-yl) -3-methoxyphenyl) azetidin-3-amine was prepared as follows.

Preparation of (6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (2-methoxy-4- (4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl) phenyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline

A mixture of (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro2H-pyran-2-yl) -6,7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (369 mg, 0.70 mmol; prepared according to example 6), bis- (pinacolato) dibor (266 mg, 1.05 mmol), [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium (P) (51 mg, 0.070 mmol) and potassium acetate (233 mg, 2.37 mmol) in dioxane (3.50 ml) were degassed and purged with nitrogen. Then the reaction mixture was heated to 80 ° C. After stirring under these conditions for 18 h, the reaction mixture was cooled to room temperature, filtered through a pad of Celite® and washed with EtOAc (100 ml). The filtrate was washed with water (100 ml), the aqueous layer was extracted with EtOAc (2x70 ml) and the combined organic layers were washed with water (3x70 ml) and saturated aqueous sodium chloride solution (50 ml) before being dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 25% ethyl acetate in hexane to give (6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -6- (2-methoxy-4- (4 , 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) -8-methyl3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro- 3H-pyrazolo [4,3-f] isoquinoline (186 mg, 46.3%) as a yellow solid.

Mass / charge: ES ⁺ [M + H] ⁺ = 576.

Preparation 4 - ((6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 6,7,8,9-tetrahydro-3H- pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenol one

An aqueous solution of hydrogen peroxide (33%; 75 μL, 0.74 mmol) was added dropwise to a stirred solution of (6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (2-methoxy-4- (4 , 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro- 3H-pyrazolo [4,3f] isoquinoline (186 mg, 0.32 mmol) and an aqueous solution of sodium hydroxide (1 M; 350 μl, 0.32 mmol) in THF (2.5 ml) at 5 ° C under air ... The resulting mixture was stirred at 5 ° C for 5 minutes. Then the reaction mixture was diluted with water (50 ml) and DCM (100 ml). The layers were separated and the aqueous layer was extracted with DCM (2x50 ml). The combined organic layers were dried over sulfate

- 70 037533 sodium, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 20 to 50% ethyl acetate in hexanes to give 4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenol (94 mg, 62.5%) in the form of a yellow solid.

Mass / charge: ES + [M + H] ⁺ = 466.

Preparation of tert-butyl-3- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1carboxylate

Diethyl (E) -diazen-1,2-dicarboxylate (88 mg, 0.20 mmol) was added to a solution of 4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- ( tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -3-methoxyphenol (94 mg, 0.20 mmol), tert α-butyl 3-hydroxyazetidine-1carboxylate (35 mg, 0.20 mmol), triphenylphosphine (53 mg, 0.20 mmol) and toluene (2.03 ml). Then the reaction mixture was heated to 110 ° C. After 15 h, the reaction mixture was cooled, concentrated under reduced pressure and purified by flash chromatography on silica, elution gradient from 0 to 30% ethyl acetate in hexanes to give tert-butyl-3- (4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline-6- yl) -3-methoxyphenoxy) azetidine-1-carboxylate (95 mg, 75%);

mass / charge: ES ⁺ [M + H] ⁺ = 621.

Preparation of trifluoroacetic acid salt and tert-butyl-3- (4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate

HCl in dioxane (4N; 383 μL, 1.53 mmol) was added dropwise to a solution of tert-butyl-3- (4 - ((6S, 8R) 7 - ((1-fluorocyclopropyl) methyl) -8-methyl- 3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate (95 mg, 0.15 mmol) in MeOH (1 ml) and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by reverse phase flash chromatography using mixtures of water (containing 0, 01% TFA) and decreasing polarity MeCN as eluents (20 to 80%) to give (6S, 8R) -6- (4 (azetidin-3-yloxy) -2-methoxyphenyl) -7 - ((1- fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (30 mg, 36%) as a TFA salt.

Ή NMR (300 MHz, METHANOL-64, 27 ° С) 0.86-1.09 (2H, m), 1.26-1.52 (3H, m), 1.59 (3H, d), 3 , 22-3.31 (1H, m), 3.36-3.47 (1H, m), 3.66 (1H, dd), 4.03 (3H, s), 4.07-4.27 (4H, m), 4.52-4.62 (2H, m), 5.14-5.27 (1H, m), 6.33 (1H, dd), 6.57-6.63 (1H , m), 6.68 (1H, d), 6.75 (1H, s), 6.93 (1H, d), 7.50 (1H, d), 8.23 (1H, s).

Indazole NH and additional H were not observed, mass / charge: ES ⁺ [M + H] ⁺ = 437.

Example 8.

N- [3,5-Difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine

- 71 037533

1-Fluoro-3-iodopropane (5.4 μl, 0.050 mmol) and diisopropylethylamine (0.013 ml, 0.070 mmol) were added to a stirred solution of N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl -7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (22 mg, 0.050 mmol ) in NMP (0.4 ml) at ambient temperature. After 16 h, the crude reaction mixture was purified directly by preparative reverse phase HPLC (Waters XB ridge C18 column, mm diameter, 100 mm long, 5 μm silica), elution gradient from 40 to 70% MeCN in water containing 0 , 2% ammonium hydroxide as a modifier. The product fractions were concentrated to dryness to obtain N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidine-3-amine (9 mg, 36%) as a pale yellow solid.

1H NMR (500 MHz, CD ₂ Cl ₂ , 27 ° C) 1.11 (3H, d), 1.69-1.78 (3H, m), 2.51-2.59 (2H, m), 2.81-2.94 (3H, m), 2.95-3.03 (1H, m), 3.16-3.30 (1H, m), 3.59-3.70 (3H, m ), 3.94-4.05 (1H, m), 4.41 (1H, t), 4.45 (1H, d), 4.51 (1H, t), 5.27 (1H, s) , 6.02 (2H, d), 6.83 (1H, d), 7.21 (1H, d), 8.03 (1H, s), 10.47 (1H, br. S);

mass / charge: ES ⁺ [M + H] ⁺ = 512.

N- (3,5-Difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3f] isoquinolin-6-yl) phenyl) azetidin-3-amine was prepared as follows.

Preparation of 2,2,2-trifluoroethyl trifluoromethanesulfonate o F3C-S-0 o ^ cf ₃

Trifluoromethanesulfonic anhydride (3.14 ml, 18.6 mmol) was added dropwise via syringe over 5 min to a stirred solution of 2,2,2-trifluoroethan-1-ol (1.23 ml, 16.9 mmol) and 2,6-dimethylpyridine (2.36 ml, 20.3 mmol) in DCM (50 ml) at -10 ° C. After 2 h, the reaction mixture was washed sequentially with aqueous HCl (1N; 2x30 ml) and saturated aqueous NaHCO ₃ (20 ml). Then, the organic layer was dried over MgSO ₄ , filtered and concentrated under reduced pressure to give 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.92 g, 23%) as a red oil.

1H NMR (300 MHz, CDCl ₃ , 27 ° C) 4.69 (2H, q).

Preparation of (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline

P ί

Hohr ^ K ^^ N ^ CFs ² I Η

2,2,2-Trifluoroethyl trifluoromethanesulfonate (1.3 g, 2.8 mmol) was added to a stirred solution of (R) -3- (2-aminopropyl) -2-methylaniline (0.460 g, 2.80 mmol; prepared according to example 3) and diisopropylethylamine (0.636 ml, 3.64 mmol) in 1,4-dioxane (10 ml). The reaction mixture was heated at 65 ° C for 15 h and then cooled and concentrated under reduced pressure. The resulting residue was dissolved in EtOAc (50 ml) and washed with a mixture of saturated aqueous NaHCO ₃ solution and saturated aqueous sodium chloride solution. The aqueous layer was extracted with EtOAc (20 ml) and the combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was dissolved in MeOH, adsorbed on diatomaceous earth under reduced pressure and purified by flash chromatography on silica, elution gradient from 1% to 6% MeOH in DCM. The product fractions were concentrated to dryness to obtain (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (0.30 g, 44%) as a pale orange resin.

1H NMR (300 MHz, CDCl ₃ , 27 ° C) 1.07 (3H, d), 2.09 (3H, s), 2.63 (1H, dd), 2.76 (1H, dd), 2 , 92-3.05 (1H, m), 3.15 (2H, q), 6.58 (2H, d), 6.85-7.00 (1H, m).

No signals were observed for the three NH groups, mass / charge: ES ⁺ [M + H] ⁺ = 247.

Obtaining (1 S, 3R) -1 - (4-bromo-2,6-difluorophenyl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1,2,3,4tetrahydroisoquinoline-6- amine

- 72 037533 (R) -2-Methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (303 mg, 1.23 mmol) and 4-bromo-2,6 difluorobenzaldehyde (544 mg, 2.46 mmol) was heated in a mixture of acetic acid (5 ml) and water (0.111 ml) at 90 ° C for 2 h. The reaction mixture was slowly cooled to ambient temperature and stirred overnight. The reaction mixture was then concentrated under reduced pressure and the resulting residue was dissolved in EtOAc (30 ml) and washed with saturated aqueous NaHCO ₃ solution (2x20 ml). The combined aqueous layers were extracted with EtOAc (2x20 ml) and the combined organic layers were washed with saturated aqueous sodium chloride solution and concentrated to minimum volume (~ 20 ml). Aqueous HCl solution (1N; 20 ml) was added and the biphasic mixture was stirred vigorously for 2 h. The layers were separated and the aqueous layer was extracted with EtOAc (20 ml). The aqueous layer was podslushivaet by adding solid sodium carbonate (to pH ~ 8, measured using a litmus strip to determine the pH) and then was extracted with DCM (3x20 ml). The combined DCM extracts were dried over MgSO ₄ , filtered and concentrated under reduced pressure to give 40 mg of the crude product. At the same time, the combined EtOAc extracts were washed with saturated aqueous sodium chloride and concentrated to dryness. The resulting residue was dissolved in THF (10 ml) and stirred with polystyrene-coupled tosylhydrazide (1.03 g, 2.76 mmol) for 14 h. The mixture was filtered and the resin was washed sequentially with THF (10 ml) and MeOH (3x10 ml ). The filtrate was combined with approximately 40 mg of the previously obtained crude product and concentrated under reduced pressure. This final residue was purified by flash chromatography on silica, elution gradient 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness to give (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -3,5-dimethyl-2- (2,2,2trifluoroethyl) -1,2,3 , 4-tetrahydroisoquinoline-6-amine (230 mg, 41%) as a pale yellow solid.

1H NMR (300 MHz, CDCl ₃ , 27 ° C) 1.04 (3H, d), 2.42 (3H, s), 2.58 (1H, dd), 2.74-2.87 (1H, m), 3.02-3.28 (2H, m), 3.46-3.59 (1H, m), 5.27 (1H, s), 6.66 (1H, d), 7.02 (2H, d), 7.32 (1H, d), 9.95-10.73 (2H, br. S);

mass / charge: ES ⁺ [M + H] ⁺ = 449.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4 , 3-f] isoquinoline

Br

A solution of sodium nitrite (35 mg, 0.51 mmol) in water (0.20 ml) was added dropwise over 1 min to a stirred solution of (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) - 3,5-dimethyl-2- (2,2,2trifluoroethyl) -1,2,3,4-tetrahydroisoquinolin-6-amine (230 mg, 0.51 mmol) in propionic acid (2 ml) at about -20 ° C (ice bath-NaCl). After 15 min, the reaction mixture was diluted with ice cold EtOAc (15 ml). The biphasic mixture was vigorously stirred under these conditions and neutralized by the slow addition of solid Na ₂ CO ₃ (to alkaline pH measured using a litmus strip). The cooling bath was removed. The phases were separated and the organic layer was washed with saturated aqueous NaHCO ₃ solution (2x15 ml), saturated aqueous sodium chloride solution (15 ml), dried over MgSO 4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 5% to 35% EtOAc in hexanes. The product fractions were concentrated to dryness to give (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8, 9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinoline (104 mg, 45%) as a pale orange film.

1H NMR (300 MHz, CDCl3, 27 ° C) 1.15 (3H, d), 2.89-3.04 (2H, m), 3.20-3.36 (1H, m), 3.50 (1H, dd), 3.613.75 (1H, m), 5.41 (1H, s), 6.80 (1H, d), 7.01-7.11 (2H, m), 7.23 ( 1H, d), 8.10 (1H, s), 10.24 (1H, br. S);

mass / charge: ES ⁺ [M + H] ⁺ = 460.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2trifluoroethyl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

Br

3,4-Dihydro-2H-pyran (0.103 ml, 1.13 mmol) and p-toluenesulfonic acid monohydrate (2 mg, 0.01 mmol) were added to a stirred solution of (6S, 8R) -6- (4-bromo- 2,6-difluorophenyl) -8-methyl-7

- 73 037533 (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (104 mg, 0.23 mmol) in DCM (1.5 ml ). The reaction mixture was maintained under reflux for 1 hour. After cooling, the reaction mixture was diluted with DCM, washed with saturated aqueous NaHCO3, dried over MgSO4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness to obtain (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -7- (2, 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (117 mg, 95%) as a pale yellow solid.

Mass / charge: ES ⁺ [M + H] ⁺ = 544.

Preparation of tert-butyl-3 - ((3,5-difluoro-4 - ((6S, 8R) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2trifluoroethyl ) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) amino) azetidine-1carboxylate

The vial was loaded with a magnetic anchor, (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -7- (2,2, 2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (55 mg, 0.10 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (26 mg , 0.15 mmol), Pd ₂ dba ₃ (9 mg, 0.01 mmol), Xantphos (12 mg, 0.02 mmol) and cesium carbonate (99 mg, 0.30 mmol). The vial was sealed and evacuated / filled with nitrogen (3x) before adding degassed 1,4-dioxane (1 ml) via syringe. The mixture was stirred at ambient temperature for 2 min, then placed in a heating block preheated to 90 ° C. After 22 h, allow the mixture to cool to room temperature and then dilute with EtOAc. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 45% EtOAc in hexanes. The product fractions were concentrated to dryness to obtain tert-butyl-3 - ((3,5 difluoro-4 - ((6S, 8R) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) amino) azetidine-1-carboxylate (46 mg, 72%) as a pale yellow solid.

¹ H NMR (300 MHz, CDCl3, 27 ° C) 1.08 (3H, d), 1.42 (9H, s), 1.55-1.81 (4H, m), 1.97-2, 23 (2H, m), 2.452.63 (1H, m), 2.80-3.02 (2H, m), 3.09-3.27 (1H, m), 3.34-3.47 ( 1H, m), 3.53-3.65 (1H, m), 3.66-3.75 (3H, m), 3.92-4.02 (1H, m), 4.19-4, 32 (3H, m), 5.25 (1H, s), 5.58-5.67 (1H, m), 5.93 (2H, dd), 6.81 (1H, d), 7.22 -7.27 (1H, m), 7.98 (1H, s).

Partial multiplet overlap at 7.22-7.27 ppm with chloroform, mass / charge: ES ⁺ [M + H] ⁺ = 636.

Receiving

N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8., 9-tetrahydro-3H pyrazolo [4, 3-f] isoquinolin-6-yl) phenyl) azetidine-3-amine

tert-Butyl-3 - ((3,5-difluoro-4 - ((6S, 8R) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2- trifluoroethyl) 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) amino) azetidine-1-carboxylate (46 mg, 0.08 mmol) was dissolved in formic acid (0.50 ml, 13 mmol) and the stirred solution was heated to 30 ° C. After 27 hours, the reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in 5% IPA / DCM and neutralized with saturated aqueous NaHCO ₃ solution. The phases were separated and the aqueous layer was extracted with 5% IPA / DCM (2x4 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure to give crude N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (38 mg, 111%), contaminated with a small amount of starting material, from which it was partially removed protection, in the form of a pale yellow film.

- 74 037533

Mass / charge: ES + [M + H] ⁺ = 452.

Example 9.

5- (6S, 8R) -7 - ((1-Fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo- [4,3f] isoquinolin-6-yl) -N - (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine

DMF (1 ml) and DIPEA (0.022 ml, 0.13 mmol) were sequentially added to the flask, which was loaded with N- (azetidin-3-yl) -5 - ((6S, 8R) -7 - ((1- fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyridin-2-amine (22 mg, 0.050 mmol). Then 1-fluoro-3-iodopropane (9 mg, 0.05 mmol) in DMF (0.1 ml) was added. After 2 h, the reaction mixture was diluted with saturated aqueous sodium chloride solution and the mixture was extracted into EtOAc (3x). The combined organic layers were washed with water and dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude film was purified by flash chromatography on silica, elution gradient from 2 to 10% MeOH in DCM, to give 5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine (9.0 mg, 36%) as a dry film.

1H NMR (500 MHz, CHLOROFORM-d, 27 ° С) δ ppm 0.46-0.66 (2H, m), 0.85-1.05 (2H, m), 1.09 (3H, d) , 1.64-1.78 (2H, m), 2.50-2.61 (3H, m), 2.83 (1H, dd), 2.87-2.96 (2H, m), 3 , 04-3.21 (2H, m), 3.56-3.69 (2H, m), 3.70-3.76 (1H, m), 3.85 (3H, s), 4.30 -4.37 (1H, m), 4.45 (2H, dt), 4.93 (1H, br d), 5.32 (1H, s), 5.82 (1H, s), 6.76 (1H, d), 7.00 (1H, d), 7.32 (1H, s), 7.99 (1H, s), 11.00 (1H, br s);

mass / charge: ES ⁺ [M + H] ⁺ = 497

N- (Azetidin-3-yl) -5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3- f] isoquinolin-6-yl) -4-methoxypyridin-2-amine was prepared as described below.

Obtaining (1 S, 3R) -1 - (6-bromo-4-methoxypyridin-3-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline-6 -amine

6-Bromo-4-methoxynicotinaldehyde (1.46 g, 6.77 mmol) was added to a solution of (R) -3- (2 - (((1fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (0.800 g, 3 , 39 mmol) in AcOH (27 ml) and water (0.305 g, 16.9 mmol) and the reaction mixture was heated at 85 ° C for 18 h. After cooling, the reaction mixture was concentrated under reduced pressure and the resulting residue was diluted with ethyl acetate and podslushivaet with using a saturated aqueous solution of sodium bicarbonate. The organic layer was combined with aqueous HCl (1N) and the biphasic mixture was stirred at room temperature for 30 minutes. The organic layer was washed with an aqueous HCl solution (1N), then the combined aqueous layers were extracted with ethyl acetate. Then the aqueous layer was podslushivaet by adding solid K ₂ CO ₃ and was extracted with ethyl acetate (2x). The combined ethyl acetate extracts from the aqueous layer, which was podslushivaet, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 10 to 100% ethyl acetate in hexanes, to give (1S, 3R) -1- (6-bromo-4-methoxypyridin-3-yl) -2 - ((1- fluorocyclopropyl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline-6-amine (0.450 g, 30.6%) as gum.

1H NMR (400 MHz, DMSO-d ₆ , 27 ° C) δ ppm 0.47-0.64 (2H, m), 0.87-0.98 (5H, m), 1.95 (3H, s ), 2.42-2.48 (1H, m), 2.79 (1H, dd), 2.87-2.99 (1H, m), 3.41-3.50 (1H, m), 3.92-3.97 (3H, m), 4.66 (2H, s), 5.11 (1H, s), 6.30 (1H, d), 6.39 (1H, d), 7 , 26 (1H, s), 7.64 (1H, s).

One hydrogen is cut off by DMSO.

Mass / charge: ES ⁺ [M + H] ⁺ = 434.

Obtaining (6S, 8R) -6- (6-bromo-4-methoxypyridin-3-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3-f] isoquinoline

- 75 037533

Br

A solution of sodium nitrite (0.045 g, 0.65 mmol) in water (0.75 ml) was added dropwise to a solution of (1S, 3R) -1- (6-bromo-4-methoxypyridin-3-yl) -2- ( (1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinolin-6-amine (0.270 g, 0.62 mmol) in propionic acid (3.0 ml) at -15 ° C (bath with salt / ice) and the reaction mixture was stirred under these conditions for 1 h. Ice-cold EtOAc (10 ml) was added followed by the addition of portions of saturated aqueous NaHCO ₃ solution (15 ml). The layers were separated and the organic layer was washed with saturated aqueous NaHCO ₃ solution (2x). The combined aqueous layers (pH 8) were extracted with EtOAc and then all the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 20 to 60% ethyl acetate in hexanes to give (6S, 8R) -6- (6-bromo-4-methoxypyridin-3-yl) -7 - (( 1 fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinoline (0.11 g, 41%) as gum.

1H NMR (400 MHz, DMSO-d6, 27 ° C) 0.47-0.64 (2H, m), 0.87-0.97 (2H, m), 0.97-1.01 (3H, m), 2.51-2.63 (1H, m), 2.89 (1H, dd), 2.94-3.06 (1H, m), 3.23 (1H, br dd), 3, 54-3.67 (1H, m), 3.97 (3H, s), 5.28 (1H, s), 6.68 (1H, d), 7.21 (1H, d), 7.30 (1H, s), 7.68 (1H, s), 8.06 (1H, s), 12.98 (1H, br s);

mass / charge: ES ⁺ [M + H] ⁺ = 445.

Preparation of (6S, SR) -6- (6-bromo-4-methoxy-3-pyridyl) -7 - [(1-fluorocyclopropyl) methyl] -8-methyl-3 [(2R) -tetrahydropyran-2-yl] -8,9-dihydro-6H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (6-6pom-4methoxypyridin-3-yl) -7 - ((1-fluorocyclopropyl) methyl) - 8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9tetrahydro-2H-pyrazolo [4,3-f] isoquinoline

Br Br

DCM (4 ml) was added to the flask, which was charged with (6S, 8R) -6- (6-bromo-4-methoxypyridine-3yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (150 mg, 0.34 mmol) and 4-methylbenzenesulfonic acid hydrate (71 mg, 0.37 mmol). 3,4-Dihydro-2H-pyran (43 mg, 0.51 mmol) was added to the stirred reaction mixture and the reaction mixture was stirred at rt overnight. The reaction mixture was washed with a saturated aqueous sodium hydrogen carbonate solution, and the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude brown gum. This was purified by flash chromatography, elution gradient from 5% to 40% EtOAc in hexanes, to give a mixture of (6S, 8R) -6- (6-bromo-4-methoxy-3-pyridyl) -7 - [(1-fluorocyclopropyl ) methyl] -8-methyl-3 - [(2R) -tetrahydropyran-2-yl] -8,9-dihydro-6H-pyrαzolo [4,3-f] isoquinoline and (6S, 8R) -6- (6 -bromo-4-methoxypyridin-3-yl) -7 ((1-fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro- 2H-pyrαzolo [4,3f] isoquinoline (151 mg, 85%) as gum.

Ή NMR (400 MHz, CHLOROFORM-d, 27 ° С) 0.45-0.61 (2H, m), 0.96-1.06 (2H, m), 1.09 (3H, d), 1.461 , 91 (3H, m), 2.03-2.20 (2H, m), 2.47-2.63 (2H, m), 2.88 (1H, ddd), 3.14 (1H, dd ), 3.20-3.31 (1H, m), 3.64-3.79 (2H, m), 3.96 (3H, d), 3.98-4.06 (1H, m), 5.40 (1H, d), 5.64-5.71 (1H, m), 6.78 (1H, d), 6.99 (1H, d), 7.25-7.33 (1H, m), 7.76 (1H, d), 8.02 (1H, d).

Preparation of tert-butyl-3 - ((5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyridin-2yl) amino) azetidine-1-carboxylate and tert-butyl-3- (5 - ((6S, 8R ) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-2 (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyridin2-ylamino) azetidine-1-carboxylate

- 76 037533

Dioxane (2.7 ml) was added to the flask, which was charged with a mixture of (6S, 8R) -6- (6-6pom-4methoxypyridin-3-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl -3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9 tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (6-bromo-4 -methoxypyridin-3-yl) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4 , 3f] isoquinoline (140 mg, 0.26 mmol), cesium carbonate (172 mg, 0.53 mmol) and tert-butyl 3-aminoazetidine 1-carboxylate (68 mg, 0.40 mmol) and evacuated from the reaction flask and filled it with nitrogen (3x). BrettPhos 3rd generation pre-catalyst (24 mg, 0.030 mmol) was added and the flask was evacuated again and filled with nitrogen (3x). The reaction mixture was heated at 110 ° C for 4 hours. The reaction mixture was cooled to room temperature, filtered and concentrated under reduced pressure. The resulting resin was purified by flash chromatography on silica, elution gradient from 30 to 100% ethyl acetate in hexanes, to give tert-6utyl-3 - ((5 - ((6S, 8R) -7 ((1-fluorocyclopopyl) methyl ) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -4-methoxypyridin-2 -yl) amino) azetidine-1-carboxylate and tert-butyl-3- (5 - ((6S, 8R) 7 - ((1-fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2H-pyran- 2-yl) -6,7,8,9-tetrahydro-2Hpyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyridin-2-ylamino) azetidine-1-carboxylate (53 mg, 32%) in the form of a dry film.

Ή NMR (400 MHz, CHLOROFORM-d, 27 ° С) 0.50-0.68 (2H, m), 0.96-1.09 (2H, m), 1.12 (3H, d), 1.421 , 47 (9H, m), 1.60-1.88 (3H, m), 2.07-2.21 (2H, m), 2.50-2.67 (2H, m), 2.85 (1H, ddd), 3.10-3.23 (2H, m), 3.633.79 (4H, m), 3.87-3.91 (3H, m), 3.99-4.07 (1H , m), 4.24-4.33 (2H, m), 4.42-4.52 (1H, m), 5.36 (1H, d), 5.645.73 (1H, m), 5, 83 (1H, d), 6.85 (1H, d), 7.26-7.32 (2H, m), 7.40 (1H, d), 8.00-8.03 (1H, m) ;

mass / charge: ES ⁺ [M + H] ⁺ = 621.

Preparation of N- (azetidin-3-yl) -5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9 tetrahydro-3H-pyrazolo [4,3 -f] isoquinolin-6-yl) -4-methoxypyridin-2-amine

Methanol (0.5 ml) was added to the flask, which was loaded with tert-butyl-3 - ((5 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro- 2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyrid in-2-yl) amino) azetidine-1 β-carboxylate (0.047 g, 0.080 mmol). HCl in dioxane (4 M; 0.5 ml, 2 mmol) was added and stirring was continued for 2 h. The reaction mixture was then concentrated under reduced pressure and the resulting residue was purified using an SCX-2 cartridge eluting with 3N. ammonia in methanol to give crude N- (azetidin-3-yl) -5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro- 3H-pyrazolo [4,3-f] isoquinolin-6-yl) -4-methoxypyridin-2-amine (25 mg, 76%) as gum. The product was used in the next step without further purification.

Mass / charge: ES ⁺ [M + H] ⁺ = 437.

Example 10.

N- (4 - ((6S, 8R) -7 - ((3- (Fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4, 3f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine

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Added 1- (3-fluoropropyl) azetidin-3-amine (47.3 mg, 0.36 mmol) in 1,4-dioxane (1.4 ml) into a sealed microwave vial containing (6S, 8R) -6- (4-bromo-2-methoxyphenyl) 7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6, 7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (100 mg, 0.18 mmol), sodium tert-butoxide (34.4 mg, 0.36 mmol) and [(2-d icyclohexylphosphino- 3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'-biphenyl)] palladium (P) methanesulfonate (BrettPhos Pd G3 ) (8.10 mg, 8.95 μmol). The vial was degassed by bubbling nitrogen for 10 minutes, then heated to 90 ° C for 1 hour. After cooling, the reaction mixture was diluted with EtOAc and washed with water. The aqueous phase was extracted with EtOAc, then the combined organics were evaporated. The crude residue was dissolved in DCM (2 ml) and TFA (1 ml) was added. The mixture was stirred for 1 hour, then evaporated. The residue was dissolved in DCM and washed with saturated aqueous NaHCO ₃ solution. The aqueous layer was extracted with DCM, then the combined organics were dried over Na ₂ SO ₄ and evaporated. The crude product was purified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5 μm silica, 19 mm in diameter, 100 mm in length) using mixtures of water (containing 0.1% NH ₃ ) and MeCN with decreasing polarity as eluents. The fractions containing the desired compound were evaporated to dryness to give N- (4 - ((6S, 8R) -7 - ((3 (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3methoxyphenyl) -1- (3-fluoropropyl) azetidine-3-amine (59.0 mg, 63%) as a beige solid substances.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.07 (3H, d), 1.68-1.87 (2H, m), 2.62 (2H, t), 2.75 (1H, d), 2.83 (1H, dd), 2.87-2.96 (3H, m), 3.10 (1H, dd), 3.27 (1H, dt), 3.74 (2H, q ), 3.84 (3H, s), 4.04-4.14 (2H, m), 4.43 (4H, td), 4.53 (1H, t), 4.58 (1H, d) , 4.78 (2H, td), 5.11 (1H, s), 5.90 (1H, dd), 6.12 (1H, d), 6.48 (1H, d), 6.80 ( 1H, d), 7.15 (1H, d), 8.05 (1H, d);

mass / charge: ES + [M + H] + = 526.

N- (4 - ((6S, 8R) -7 - ((3- (Fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4, 3f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine was prepared as follows.

Obtaining (3- (fluoromethyl) oxetan-3-yl) methanol

Cesium fluoride (5.03 g, 33.14 mmol) was added to a flask containing (3- (bromomethyl) oxetane-3yl) methanol (2.00 g, 11.05 mmol) in ethylene glycol (6 mL) and the reaction mixture was heated to 150 ° C for 2 h. After cooling, the reaction mixture was diluted with EtOAc (30 ml) and water (30 ml). The aqueous layer was extracted with diethyl ether (3x30 ml) and ethyl acetate (3x30 ml). The combined organics were dried over MgSO ₄ , filtered and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 100% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give (3- (fluoromethyl) oxetan-3-yl) methanol (0.734 g, 55%) as a colorless liquid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 3.92 (2H, s), 4.52 (4H, t), 4.59-4.89 (2H, m). Preparation of (3- (fluoromethyl) oxetan-3-yl) methyl trifluoromethanesulfonate

To a solution of (3- (fluoromethyl) oxetan-3-yl) methanol (1.00 g, 8.32 mmol) in DCM (30.7 ml) was added trifluoromethanesulfonic anhydride (1.47 ml, 8.74 mmol) with followed by the addition of 2,6-dimethylpyridine (1.12 ml, 9.57 mmol) and the reaction mixture was stirred at 0 ° C for 1 h. The reaction mixture was washed with water and 1N. citric acid solution, then dried over Na ₂ SO ₄ , filtered and evaporated to give (3- (fluoromethyl) oxetan-3-yl) methyl trifluoromethanesulfonate (1.98 g, 94%) as a red oil, which was used directly without further purification ...

Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 4.40-4.75 (8H, m).

Preparation of (R) -3- (2 - (((3- (fluoromethyl) oxetan-3-yl) methyl) amino) propyl) -2-methylaniline (3- (Fluoromethyl) oxetan-3-yl) methyl trifluoromethanesulfonate (1, 92 g, 7.61 mmol) was added to a solution of (R) -3- (2-aminopropyl) -2-methylaniline (1.00 g, 6.09 mmol) and DIPEA (1.58 mL, 9.13 mmol) in 1,4-dioxane (18.7 ml) and the reaction mixture was heated to 70 ° C for 5 h. After cooling, the reaction mixture was diluted with EtOAc and washed with water. The aqueous phase was extracted with EtOAc, then the combined organics were dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 10% MeOH in EtOAc. The pure fractions were evaporated to dryness to give (R) -3- (2 - (((3- (fluoromethyl) oxetan-3-yl) methyl) amino) propyl) -2-methylaniline (0.673 g, 42 %) in the form of a pale yellow resin.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.11 (3H, d), 2.04 (3H, s), 2.59-2.71 (1H, m), 2.76-3.10 (4H, m), 4.344.52 (4H, m), 4.53-4.68 (2H, m), 6.58 (1H, d), 6.60 (1H, d), 6.95 ( 1H, t);

mass / charge: ES ⁺ [M + H] ⁺ = 267.

Preparation of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline -6-amine

Br

4-Bromo-2-methoxybenzaldehyde (645 mg, 3.00 mmol) was added to a solution of (R) -3- (2 - (((3 (fluoromethyl) oxetan-3-yl) methyl) amino) propyl) -2- methylaniline (400 mg, 1.5 mmol) in acetic acid (7.4 ml) and water (135 μl, 7.50 mmol). The reaction mixture was heated to 80 ° C for 4 hours. After cooling, the acetic acid was evaporated, then the residue was dissolved in DCM and washed with saturated aqueous NaHCO ₃ solution. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated. The residue was dissolved in methanol (5 ml), then hydroxylamine hydrochloride (313 mg, 4.50 mmol) and potassium acetate (588 mg, 6.00 mmol) were added and the reaction mixture was stirred at room temperature for 30 min. The volatiles were evaporated, then the residue was taken up in DCM and water. The layers were separated, then the aqueous layer was extracted with DCM. The combined organics were dried over Na ₂ SO ₄ , filtered and evaporated, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give (1S, 3R) -1- (4-6pom-2methoxyphenyl) -2 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -3,5-dimethyl -1,2,3,4-tetrahydroisoquinoline-6amine (302 mg, 43%) as a beige solid.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.01 (3H, d), 2.07 (3H, s), 2.45 (1H, dd), 2.61-2.73 (1H, m ), 2.86 (1H, d), 3.01-3.14 (1H, m), 3.54 (2H, s), 3.87 (3H, s), 4.41-4.48 ( 2H, m), 4.51 (1H, d), 4.53-4.60 (2H, m), 4.67 (1H, d), 4.73-4.81 (1H, m), 5 , 00 (1H, s), 6.47 (1H, s), 6.47 (1H, s), 6.61 (1H, d), 6.88 (1H, dd), 7.01 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 463.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl6,7,8,9-tetrahydro-3H -pyrαzolo [4,3-f] isoquinoline

Br

Sodium nitrite (39.1 mg, 0.57 mmol) in water (0.5 ml) was added to a cooled solution of (1S, 3R) -1 (4-bromo-2-methoxyphenyl) -2 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (250 mg, 0.54 mmol) in propionic acid (2.2 ml) at -15 ° C (dry ice / acetone bath). After stirring for 30 min, ice-cold toluene (15 ml) was added and the reaction mixture was stirred at 0 ° C for 15 min. and then warmed to room temperature over 1 h. Water (15 ml) was added and the layers were separated. The aqueous layer was extracted with EtOAc (2x15 ml), then the combined organics were washed with saturated aqueous sodium chloride solution, dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (188 mg, 74%) as a beige solid.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.07 (3H, d), 2.70 (1H, d), 2.87 (1H, dd), 2.97 (1H, d), 3, 11-3.18 (1H, m), 3.18-3.27 (1H, m), 3.90 (3H, s), 4.42-4.52 (3H, m), 4.59 ( 1H, d), 4.63-4.87 (2H, m), 5.19 (1H, s), 6.63 (1H, d), 6.76 (1H, d), 6.88 (1H , dd), 7.05 (1H, d), 7.19 (1H, d), 8.09 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 474.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2 -yl) -6,7,8,9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinoline

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3,4-Dihydro-2H-pyran (67.3 μL, 0.74 mmol) was added to a solution of (6S, 8R) -6- (4-6pom-2methoxyphenyl) -7 - ((3- (fluoromethyl) oxetane- 3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (175 mg, 0.37 mmol) and 4-methylbenzenesulfonic acid hydrate (14.0 mg , 0.07 mmol) in DCM (3.6 ml) and the reaction mixture was stirred at 40 ° C for 2 h. After cooling, the reaction mixture was diluted with DCM and washed with saturated aqueous NaHCO ₃ solution, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was passed through a silica plug eluting with EtOAc / heptane (1: 1). The filtrate was evaporated to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-3- (tetrahydro2H-pyran -2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (196 mg, 95%) as a beige solid, which was used directly in the next step.

Mass / charge: ES + [M + H] ⁺ = 558.

Example 11.

N- (3,5-Difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f ] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine

4n. HCl in dioxane (0.25 ml, 0.99 mmol) was added to a solution of N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidine-3- amine (58 mg, 0.10 mmol) in methanol (0.30 ml) and the reaction mixture was stirred at room temperature for 2 h. The volatiles were evaporated and the residue was suspended in saturated aqueous NaHCO ₃ solution (30 ml) and extracted with DCM (x2). The organic phase was dried over MgSO ₄ , filtered and evaporated. The crude residue was purified by chromatography on silica and eluted using a gradient of 0 to 10% 1M NH ₃ / MeOH in DCM. The fractions containing the product were evaporated to dryness to give N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8, 9 tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine (45.4 mg, 91%) as a white solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.47 (2H, dtd), 0.95 (2H, dt), 1.08 (3H, d), 1.68-1.85 (2H, m), 2.59 (2H, s), 2.69 (1H, dd), 2.91 (3H, dq), 3.14 (1H, dd), 3.43 (1H, dd), 3, 69 (2H, q), 3.81 (1H, dd), 4.01 (1H, q), 4.28 (1H, d), 4.43 (1H, t), 4.53 (1H, t ), 5.19 (1H, s), 5.96 (2H, d), 6.82 (1H, d), 7.15 (1H, d), 8.06 (1H, d), 10.36 (1H, s);

mass / charge (ES ⁺ ) [M + H] ⁺ = 502.

N- (3,5-Difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3 -f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine was prepared as follows.

Preparation of (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline-6-amine

Bg

1LJL IF (R) -3- (2 - (((1-Fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (0.35 g, 1.48 mmol) and 4-bromo2,6-difluorobenzaldehyde (0, 33 g, 1.48 mmol) in a mixture of H ₂ O (0.13 ml, 7.40 mmol) and acetic acid (5.79 ml) was heated at 80 ° C for 7 h. The solvent was evaporated and the residue was treated with HCl (1N, 10 ml) and stirred at room temperature for 1 hour. Solid Na ₂ CO _{3 was} added to the reaction mixture until the pH of the reaction mixture was> 8. The mixture was diluted with H ₂ O (40 ml) and was extracted twice with EtOAc (2x50 ml). The organic phase was dried over MgSO ₄ , filtered, concentrated and purified by chromatography on silica (10 to 70% EtOAc in heptanes) to give (1S, 3R) -1- (4-bromo-2,6- difluorophenyl) -2 - ((1fluorocyclopropyl) methyl) -3,5-imethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (0.285 g, 44%) as a pale yellow gum.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.44 (2H, dq), 0.91-0.95 (1H, m), 0.95-0.99 (1H, m), 1, 02 (3H, d), 2.06 (3H, s), 2.50-2.65 (2H, m), 3.00-3.15 (2H, m), 3.50 (2H, s) , 3.69 (1H, d), 5.17 (1H, s), 6.42 (2H, s), 6.97 (1H, d), 6.99 (1H, d);

mass / charge (ES ⁺ ) [M + H] ⁺ = 439.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3Н-pyrazolo [4, 3-f] isoquinoline (1S, 3R) -1- (4-Bromo-2,6-difluorophenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline- 6-amine (0.280 g, 0.64 mmol) in propionic acid (2.66 ml) was cooled to -17 ° C (dry ice / acetone bath). Sodium nitrite (0.044 g, 0.64 mmol) in water (0.53 ml) was added dropwise and the reaction mixture was stirred at -17 ° C for 30 min. The reaction mixture was diluted with ice-cold toluene (15 ml) and stirred at 4 ° C for 15 min. The mixture was then stirred and warmed to room temperature over 45 minutes. The reaction mixture was washed with water (2x15 ml), the combined aqueous phases were washed with EtOAc (2x10 ml), the combined organics were washed with saturated aqueous sodium chloride solution (1x20 ml), dried (MgSO ₄ ), filtered and the filtrate was evaporated to orange brown oil. The crude material was purified by flash chromatography on silica, gradient elution 0-40% EtOAc in heptane to give (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1-fluorocyclopropyl ) methyl) -8-methyl6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.216 g, 75%) as an off-white solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.46 (2H, dddd), 0.91-1.04 (2H, m), 1.07 (3H, d), 2.66 (1H, dd), 2.95 (1H, dd), 3.15 (1H, dd), 3.46 (1H, dd), 3.83 (1H, dd), 5.32 (1H, s), 6, 76 (1H, d), 7.00 (2H, d), 7.18 (1H, d), 8.09 (1H, d), 10.52 (1H, s);

mass / charge (ES ⁺ ) [M + H] ⁺ = 450.

Obtaining (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (4-6pom-2,6difluorophenyl) -7 - ((1-fluorocyclopropyl) methyl) - 8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3-f] isoquinoline

(6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1 fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro- 3H-pyrαzolo [4,3-f] isoquinoline (216 mg,

0.48 mmol), 4-methylbenzenesulfonic acid hydrate (9.1 mg, 0.05 mmol), 3,4-dihydro-2H-pyran (0.07 ml, 0.72 mmol) and DCM (2 ml). The mixture was heated in a microwave oven at 80 ° C for 20 minutes. An additional amount (0.035 ml, 0.36 mmol) of 3,4-dihydro-2H-pyran was added and the reaction mixture was heated to 85 ° C for an additional 15 min. The reaction mixture was diluted with DCM and washed with saturated aqueous NaHCO ₃ , then the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give two regioisomeric THP-protected products, which are (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1fluorocyclopropyl) methyl) -8-methyl -3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline and (6S, 8R) -6- (4-bromo-2, 6-difluorophenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-2 (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3 -f] isoquinoline (237 mg, 92%), as a pale yellow gum (data shown for the major isomer).

- 81 037533

Ή NMR (500 MHz, CDC13, 27 ° С) 0.35-0.55 (2H, m), 0.96 (2H, ddd), 1.05 (3H, dd), 1.58-1.84 (3H, m),

1.97-2.32 (3H, m), 2.64 (1H, dd), 2.77 (1H, dd), 3.14 (1H, ddd), 3.29-3.54 (1H, m), 3.62-3.91 (2H, m), 3.96-4.25 (1H, m), 5.26 (1H, d), 5.65 (1H, ddd), 6.64 (1H, d), 7.00 (2H, d), 7.26-7.45 (1H, m), 8.13 (1H, dd);

mass / charge (ES +) [M + H] ⁺ = 534.

Preparation of N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2Hpyran-2-yl) -6.7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) αzetidin-3 amine

[(2-Dicyclohexylphosphino-3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'biphenyl)] palladium (P) methanesulfonate (BrettPhos Pd G3) (5.0 mg, 5.54 μmol) was added to a suspension of (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (4- bromo-2,6-difluorophenyl) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3f] isoquinoline (37 mg, 0.07 mmol), 1- (3-fluoropropyl) azetidine-3-amine (22.9 mg, 0.17 mmol) and sodium tert-butoxide (13.3 mg, 0.14 mmol) in degassed 1,4-dioxane (0.58 ml) and the reaction mixture was heated to 95 ° C for 1.5 h in a microwave oven. The reaction mixture was diluted with EtOAc, filtered through celite and evaporated to dryness. The crude product was purified by flash chromatography on silica, gradient elution 0 to 10% 1M NH ₃ / MeOH in DCM. The pure fractions were evaporated to dryness to give N- (3,5-difluoro-4 ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-2- (tetrahydro-2H-pyran-2 -yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) αzetidin-3-amine (18.0 mg, 44 %) in the form of a colorless dry film.

1H NMR (500 MHz, CDCl3, 27 ° C) 0.38-0.54 (2H, m), 0.94 (2H, ddd), 1.06 (3H, dd), 1.67-1.82 (4H, m), 2.04-2.25 (4H, m), 2.59 (2H, t), 2.63-2.77 (2H, m), 2.89 (2H, dd), 3.14 (1H, s), 3.29-3.37 (1H, m), 3.70 (2H, q), 3.75-3.81 (2H, m), 4.00 (1H, q), 4.12-4.17 (1H, m), 4.22 (1H, d), 4.43 (1H, t), 4.52 (1H, t), 5.10 (1H, s ), 5.555.74 (1H, m), 5.96 (2H, d), 6.71 (1H, d), 7.28-7.44 (1H, m), 8.10 (1H, dd) ;

mass / charge (ES +) [M + H] ⁺ = 586.

Examples 12 and 13.

Preparation of individual diastereoisomers (6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -6- (4- (1- (3fluoropropyl) azetidin-3-yloxy) -2-methoxyphenyl) -8- methyl 6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline

Λ JO '' (UM

ADA / ^F

HN γ HN J

Into the flask, which was loaded with (6S, 8R) -6- (4- (azetidin-3-yloxy) -2-methoxyphenyl) -7- (2-fluoro-3methoxy-2-methylpropyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (32 mg, 0.07 mmol), DMF (0.66 ml) was added followed by the addition of DIPEA (23.9 μl, 0, 14 mmol). To the stirred reaction mixture, 1-fluoro-3-iodopropane (13.5 mg, 0.07 mmol) was added dropwise and the reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness and the residue was purified by flash chromatography on silica, elution gradient from 0 to 10% 1 M NH ₃ / MeOH in DCM. The fractions containing the product were evaporated to dryness to give the product as a diastereoisomeric mixture. Diastereoisomers were separated by chiral preparative SFC (Phenomonex Lux C4 column, 5 μm silica, 30 mm diameter, 250 mm long) using an isocratic mixture of 40% MeOH + 0.1% NH ₃ in CO ₂ with decreasing concentration as eluent to obtain the first eluted isomer (8.0 mg, 22%) and the second eluted isomer (8.0 mg, 22%) as pale yellow dry films.

Isomer 1: 1H NMR (500 MHz, CDO3, 27 ° C) 1.06 (3H, d), 1.28 (3H, d), 1.72-1.83 (2H, m), 2.47 ( 1H, dd), 2.63 (2H, t), 2.8-2.95 (2H, m), 3.03-3.11 (2H, m), 3.19 (2H, s), 3 , 33 (3H, s), 3.56 (1H, dd), 3.67 (1H, d), 3.79 (2H, q), 3.86 (3H, s), 4.44 (1H, t), 4.53 (1H, t), 4.74 (1H, t), 5.32 (1H, s), 6.10 (1H, dd), 6.41 (1H, d), 6, 69 (1H,

- 82 037533

d), 6.76 (1H, d), 7.13 (1H, d), 8.05 (1H, d), 10.20 (1H, s);

mass / charge (ES ⁺ ) [M + H] ⁺ = 529;

Isomer 2: ¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.06 (3H, d), 1.25 (3H, d), 1.70-1.82 (2H, m), 2, 45-2.56 (1H, m), 2.63 (2H, t), 2.78-2.91 (2H, m), 3.02-3.11 (2H, m), 3.18 ( 1H, d), 3.36 (3H, s), 3.42 (1H, dd), 3.47-3.55 (1H, m), 3.62 (1H, d), 3.74-3 , 82 (2H, m), 3.86 (3H, s), 4.43 (1H, t), 4.53 (1H, t), 4.73 (1H, t), 5.36 (1H, s), 6.08 (1H, dd), 6.41 (1H, d), 6.67 (1H, d), 6.79 (1H, d), 7.15 (1H, d), 8, 06 (1H, d), 10.20 (1H, s);

mass / charge (ES ⁺ ) [M + H] ⁺ = 529.

(6S, 8R) -6- (4- (Azetidin-3-yloxy) -2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl 6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinoline was prepared as follows.

Preparation of a diastereoisomeric mixture (6S, 8R) -6- (4- (azetidin-3-yloxy) -2-methoxyphenyl) -7- (2fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

tert-Butyl 3-hydroxyazetidine-1-carboxylate (102 mg, 0.59 mmol), (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2-fluoro-3-methoxy- 2-methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3- £] isoquinoline (110 mg, 0.20 mmol ), tert-butyl 3-hydroxyazetidine-1-carboxylate (102 mg, 0.59 mmol), 3rd generation Pd RockPhos (16.6 mg, 0.02 mmol) and cesium carbonate (128 mg, 0.39 mmol) was suspended in toluene (1.26 ml) and sealed in a microwave tube. The reaction mixture was heated to 95 ° C for 20 h. The reaction mixture was diluted with EtOAc (25 ml) and filtered through celite. The crude product was purified by flash chromatography on silica with a gradient elution of 0-100% EtOAc in heptane. The fractions containing the contaminated product were evaporated to dryness and dissolved in DCM (1.3 ml) to which TFA (378 μl, 4.91 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 2 hours, then the volatiles were removed in vacuo. The residue was washed with saturated aqueous NaHCO ₃ solution (30 ml) and was extracted with DCM (2x30 ml). The combined organic phase was dried over MgSO ₄ , filtered and evaporated. The crude product was purified by flash chromatography on silica, gradient elution 0 to 40% 1M NH ₃ / MeOH in DCM. Pure fractions were evaporated to dryness to give (6S, 8R) -6- (4 (azetidin-3-yloxy) -2-methoxyphenyl) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl -6,7,8,9-tetrahydro3H-pyrazolo [4.3- £] isoquinoline (35.0 mg, 38%) as a white solid.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.06 (3H, dd), 1.24-1.35 (3H, m), 2.49 (1H, dt), 2.76-2.96 (2H, m), 3.08-3.27 (2H, m), 3.34 (3H, d), 3.41 (1H, s), 3.53-3.69 (2H, m), 3.69-3.93 (7H, m), 4.81-5.08 (1H, m), 5.34 (1H, d), 6.02-6.12 (1H, m), 6, 40 (1H, t), 6.68 (1H, s), 6.77 (1H, dd), 7.13 (1H, d), 8.05 (1H, t);

mass / charge (eS ⁺ ) [M + H] ⁺ = 469.

Examples 14 and 15.

Preparation of individual diastereoisomers N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3 -f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine

1- (3-Fluoropropyl) azetidin-3-amine (175 mg, 1.32 mmol), (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2fluoro-3-methoxy-2 -methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (370 mg, 0.66 mmol) and sodium tert-butoxide (127 mg, 1.32 mmol) were suspended in 1,4-dioxane (8 ml). The mixture was degassed and [(2-dicyclohexylphosphino-3,6dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'-biphenyl)] palladium was added (P) methanesulfonate (Brett Phos G3) (60 mg, 0.07 mmol). The reaction mixture was heated to 100 ° C for 3 h. The reaction mixture was diluted with DCM (100 ml) and washed with water (100 ml). The organic layer was evaporated. This method was repeated using 114 mg (0.3 mmol) of aryl bromide. The combined crude products from these reactions were suspended in hydrochloric acid (2M, 5 ml) and stirred at room temperature for 2 hours.

- 83 037533 was added with water (10 ml) and washed with EtOAc (10 ml). The aqueous phase was podslushivaet using

M aqueous sodium hydroxide solution and extracted with DCM (2x15 ml). The combined organics were evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in DCM. The fractions were evaporated to dryness to give the crude product as a mixture of diastereoisomers, which were separated by chiral preparative SFC (Phenomonex Lux C4 column, 5 μm silica, 30 mm in diameter, 250 mm in length) using an isocratic mixture of 40% MeOH + 0 , 1% NH ₃ in CO ₂ with decreasing concentration as eluent to give the first eluted isomer (42 mg, 19%) and the second eluted isomer (29 mg, 13%).

Isomer 1: 1H NMR (500 MHz, DMSO, 27 ° C) 0.96 (3H, d), 1.20 (3H, d), 2.36-2.44 (1H, m), 2.65- 2.85 (3H, m), 2.92 (4H, q), 3.05-3.22 (3H, m), 3.23 (3H, s), 3.24-3.27 (2H, m), 3.46-3.59 (2H, m), 3.78 (3H, s), 4.49 (2H, dd), 5.16 (1H, s), 5.35 (1H, t ), 5.95 (1H, dd), 6.23 (1H, d), 6.36 (1H, d), 6.63 (1H, d), 7.16 (1H, d), 8.02 (1H, s), 12.90 (1H, s);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -219.77, -149.11 mass / charge: ES ⁺ [M + H] ⁺ = 528.

Isomer 2: 1H NMR (500 MHz, DMSO, 27 ° C) 0.97 (3H, d), 1.19 (3H, d), 2.68-2.81 (3H, m), 2.88- 2.95 (3H, m), 3.05-3.11 (1H, m), 3.21-3.28 (5H, m), 3.32-3.39 (1H, m), 3, 43-3.56 (2H, m), 3.79 (3H, s), 4.49 (2H, dd), 5.17 (1H, s), 5.34 (1H, t), 5.93 (1H, dd), 6.23 (1H, d), 6.35 (1H, d), 6.64 (1H, d), 7.16 (1H, d), 8.02 (1H, s) , 12.90 (1H, s);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -219.76, -148.20;

mass / charge: ES ⁺ [M + H] ⁺ = 528.

Example 16.

Preparation of 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8methyl-3,6 , 8,9-tetrahydro-7Н-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol

Hydrochloric acid (2.0 M, 0.5 ml) was added to 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -3,6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propane -1-olu (0.132 g, 0.23 mmol) and the solution was stirred at room temperature for 3 hours. The reaction mixture was purified by ion exchange chromatography using an SCX column. The desired product was eluted from the column using 1 M NH ₃ / MeOH to give 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol (0.084 g, 75% ) as a white solid.

1H NMR (500 MHz, DMSO, 27 ° C) 1.04 (3H, d), 1.20-1.28 (2H, m) 1.59-1.71 (2H, m), 2.60- 2.70 (1H, m), 2.73-2.84 (3H, m), 2.99 (1H, dd), 3.08-3.18 (1H, m), 3.41 (1H, d), 3.61-3.72 (4H, m), 3.93 (1H, d), 4.44 (2H, dt), 4.94 (1H, s), 5.42 (1H, t ), 6.21 (1H, d), 6.78-6.82 (2H, m), 6.90 (1H, d), 7.21 (1H, d), 7.74 (1H, d) , 8.03 (1H, s); 12.94 (1H, s);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -218.24, -108.31;

mass / charge: ES ⁺ [M + H] ⁺ = 489.

2,2-Difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3 ( tetrahydro-2H-pyran-2-yl) -3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol was prepared as follows.

Obtaining (1S, 3R) -1- (5-bromopyridin-2-yl) -2- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -3,5-dimethyl-1,2, 3,4-tetrahydroisoquinoline-6-amine

(R) -3- (2 - ((3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyl) amino) propyl) -2-methylaniline (1.20 g, 2.42 mmol) and 5- bromopicolinaldehyde (0.944 g, 5.07 mmol) was heated in acetic acid (12 ml) and water (0.22 ml, 12.08 mmol) to 70 ° C for 30 min. The reaction mixture was evaporated and the residue was redissolved in ethanol (10 ml). Potassium acetate (0.594 g, 6.05 mmol) and hydrochloride hydrochloride were added

- 84 037533 xylamine (0.252 g, 3.63 mmol). The reaction mixture was stirred at room temperature for 30 min, then evaporated and the residue was partitioned between DCM (30 ml) and saturated aqueous sodium bicarbonate solution (30 ml). The organic phase was evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane to give (1S, 3R) -1- (5-bromopyridin-2-yl) -2- ( 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (1.090 g, 68%) as a colorless gum.

1H NMR (500 MHz, DMSO, 27 ° C) 0.96 (9H, s), 0.98 (3H, d), 1.94 (3H, s), 2.40 (1H, dd), 2, 57 (1H, dd), 2.66-2.78 (1H, m), 3.11-3.22 (2H, m), 3.84-3.93 (1H, m), 3.95- 4.05 (1H, m), 4.69 (2H, s), 4.91 (1H, s), 6.39 (1H, d), 6.42 (1H, d), 6.99 (1H , d), 7.39-7.48 (6H, m), 7.59 (4H, ddd), 7.80 (1H, dd), 8.51 (1H, dd);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -109.84 (d), -108.39 (d);

mass / charge: ES ⁺ [M + H] ⁺ = 664/666.

Preparation of (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

Br

Sodium nitrite (119 mg, 1.72 mmol) in water (1.0 ml) was added to an ice-cold solution of (1S, 3R) -1- (5-bromopyridin-2-yl) -2- (3 - ((tert -butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline-6-amine (1.09 g, 1.64 mmol) and p-toluenesulfonic acid hydrate (0, 94 g, 4.92 mmol) in acetonitrile (6 ml). The reaction mixture was stirred for 15 minutes and then warmed to room temperature over 45 minutes. After cooling back to 4 ° C, tetrabutylammonium acetate (2.54 g, 8.20 mmol) in ice cold MeCN (40 mL) was added and the reaction mixture was stirred for 15 min before warming to room temperature for an additional 30 min, EtOAc ( 120 ml) followed by the addition of 2 M NaOH (100 ml). The organic phase was washed with saturated aqueous sodium chloride solution, dried (Na ₂ SO ₄ ) and evaporated. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (3 - ((tert -butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.330 g, 30%) as light brown foam.

¹ H NMR (500 MHz, DMSO, 27 ° C) 0.97 (9H, s), 1.06 (3H, d), 2.75-2.91 (2H, m), 2.99-3, 06 (1H, m), 3.343.41 (2H, m), 3.88-4.01 (2H, m), 5.12 (1H, s), 6.83 (1H, d), 7.12 (1H, d), 7.26 (1H, d), 7.39-7.45 (4H, m), 7.467.50 (2H, m), 7.56-7.62 (4H, m), 7.85 (1H, dd), 8.07-8.09 (1H, m), 8.54 (1H, dd), 13.02 (1H, s);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -110.01 (d), -108.72 (d);

mass / charge: ES ⁺ [M + H] ⁺ = 675/677.

Preparation of a diastereoisomeric mixture (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-3- (tetrahydro -2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3 -f] isoquinoline

Br

To a solution of (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.330 g, 0.49 mmol) and 3,4-dihydro2H-pyran (0.13 ml, 1.47 mmol) in DCM (5 ml) p-toluenesulfonic acid hydrate (93 mg, 0.49 mmol) was added and the reaction mixture was heated at 40 ° C for 1 h. The reaction mixture was diluted with DCM (20 ml) and washed with saturated aqueous sodium bicarbonate solution (20 ml ). The organic phase was evaporated to a dark brown oil and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane to give (6S, 8R) -6- (5-bromopyridin-2-yl ) -7- (3 - ((tert-butyldiphenylsilyl) oxy) 2,2-difluoropropyl) -8-methyl-3 - ((S) -tetrahydro-2H-pyran-2-yl) -6,7,8, 9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (0.330 g, 89%) as gum.

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -110.03 (d), -108.83 (d), -108.79 (d);

mass / charge: ES ⁺ [M + H] ⁺ 759/761.

Preparation of a diastereoisomeric mixture 6 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2,2difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-1] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

- 85 037533

1- (3-Fluoropropyl) azetidin-3-amine (0.115 g, 0.87 mmol), (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (3 ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.330 g , 0.43 mmol), dicyclohexyl (2 ', 4', 6'-triisopropyl-3,6dimethoxy [1,1'-biphenyl] -2-yl) phosphane (0.023 g, 0.04 mmol) and tert-butoxide sodium (0.125 g, 1.30 mmol) was suspended in 1,4-dioxane (4 ml). The mixture was degassed and [(2-dicyclohexylphosphino-3,6dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'-biphenyl)] palladium was added (P) methanesulfonate (BrettPhos G3) (0.039 g, 0.04 mmol). The reaction mixture was heated to 100 ° C for 3 h. The reaction mixture was diluted with DCM (50 ml) and washed with water (50 ml). The organic layer was evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in DCM, to give 6 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy ) -2,2difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) - N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine (0.264 g, 75%) as a yellow gum as a mixture of diastereoisomers.

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -218.19, -110.13, -109.44, -108.6 -107.8;

mass / charge: ES + [M + H] ⁺ 811.

Preparation of a diastereoisomeric mixture of 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3yl) amino) pyridin-2-yl) -8-methyl-3 , 6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol

Tetrabutylammonium fluoride (1.0 M in THF, 0.5 ml, 0.50 mmol) was added to a solution of 6 - ((6S, 8R) -7- (3 ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl ) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine (0.264 g, 0.33 mmol) in THF (2 ml). The reaction mixture was stirred at room temperature for 4 h, the reaction mixture was evaporated and the residue was partitioned between DCM (20 ml) and water (20 ml). The organic phase was evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in DCM, to give 2,2-difluoro-3 - ((6S, 8R) 6- (5 - (( 1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 3,6,8,9-tetrahydro-7H -pyrazolo [4,3-1] isoquinolin-7-yl) propan-1-ol (0.132 g, 71%) as a colorless dry film in the form of a mixture of diastereoisomers.

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -218.19, -108.42, -108.39;

mass / charge: ES ⁺ [M + H] ⁺ = 573.

Example 17.

Preparation of N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro -3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine

1- (3-Fluoropropyl) azetidin-3-amine (50 mg, 0.38 mmol), (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-3 (tetrahydro-2H- pyran-2-yl) -7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-G] isoquinoline (0.100 g, 0.20 mmol), cesium carbonate (128 mg, 0.39 mmol) and [(2-dicyclohexylphosphino-3,6-dimethoxy2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1 , 1'-biphenyl)] palladium (II) methanesulfonate (Brett Phos G3) (23 mg, 0.02 mmol) was suspended in 1,4-dioxane (2 ml) and sealed in a microwave tube. The reaction mixture was heated to 100 ° C for 4 hours under the influence of microwave radiation. The reaction mixture was diluted with DCM (20 ml) and washed with water (20 ml). The organic layer was evaporated and the crude product was dissolved in DCM (3 ml) and TFA (0.3 ml). Reak

86 037533 The reaction mixture was stirred at room temperature for 1 h, then the reaction mixture was partitioned between DCM (20 ml) and 2 M NaOH (20 ml). The organic phase was evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 20% MeOH in DCM. The fractions containing the product were evaporated to dryness to give N- (1 (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl ) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (9.0 mg, 10%) as a colorless dry film.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 1.70-1.82 (2H, m), 2.62 (2H, t), 2.85 (1H, dd), 2.93-3.06 (3H, m), 3.20-3.31 (2H, m), 3.59 (1H, td), 3.73 (2H, dt), 4.10 (2H, dd), 4.48 (2H, dt), 5.03 (1H, s), 6.80 (1H, dd), 6.89 (1H, d), 7.15 (1H, d) , 7.21 (1H, d), 7.84 (1H, dd), 8.01 (1H, d);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) -224.69, -75.55;

mass / charge: ES + [M + H] + = 477.

(6S, 8R) -6- (5-Bromopyridin-2-yl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2-trifluoroethyl) 6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline was prepared as follows.

Preparation of (1S, 3R) -1- (5-bromopyridin-2-yl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1,2,3,4-tetrahydroisoquinolin-6-amine

Bg

(R) -2-Methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (0.290 g, 1.18 mmol) and 5-bromopicolinaldehyde (0.460 g, 2.47 mmol) heated in acetic acid (6 ml) and water (0.1 ml) to 70 ° C for 30 min. The reaction mixture was evaporated and the residue was dissolved in ethanol (10 ml). Potassium acetate (0.290 g, 2.95 mmol) and hydroxylamine hydrochloride (0.123 g, 1.77 mmol) were added and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was evaporated and the residue was partitioned between DCM (30 ml) and saturated aqueous sodium bicarbonate solution (30 ml). The organic phase was evaporated and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane to give (1S, 3R) -1- (5-bromopyridin-2-yl) -3,5-dimethyl -2- (2,2,2-trifluoroethyl) -1,2,3,4-tetrahydroisoquinoline-6-amine (0.264 g, 54%) as a colorless dry film.

1H NMR (500 MHz, DMSO, 27 ° C) 1.03 (3H, d), 1.94 (3H, s), 2.45 (1H, dd), 2.67 (1H, dd), 2, 90 (1H, dd), 3.22-3.29 (1H, m), 3.49 (1H, dd), 4.69 (2H, s), 4.85 (1H, s), 6.40 -6.46 (2H, m), 7.21 (1H, d), 7.95 (1H, dd), 8.56 (1H, dd);

¹⁹ F NMR (471 MHz, DMSO, 27 ° C) 69.83;

mass / charge: ES ⁺ [M + H] ⁺ = 414/416.

Obtaining (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2trifluoroethyl) -6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline Br

Sodium nitrite (0.048 g, 0.70 mmol) in water (0.2 ml) was added to a cooled solution of (1S, 3R) -1 (5-bromopyridin-2-yl) -3,5-dimethyl-2- (2 , 2,2-trifluoroethyl) -1,2,3,4-tetrahydroisoquinolin-6-amine (0.264 g, 0.64 mmol) in propionic acid (2.5 ml) at -10 ° C. The reaction mixture was stirred for 1 h, then ice-cold EtOAc (10 ml) was added. The reaction mixture was quenched by the addition of an aqueous saturated sodium bicarbonate solution (15 ml) and stirred for 15 min before being allowed to warm to room temperature. The organic phase was dried (Na ₂ SO ₄ ) and evaporated to give crude (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) 6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.272 g, 0.64 mmol) as a brown gum. The material was dissolved in DCM (10 ml) and p-toluenesulfonic acid hydrate (0.012 g, 0.06 mmol) was added and the mixture was heated at 40 ° C for 2 h. The reaction mixture was diluted with DCM (20 ml) and washed with aqueous NaOH solution (2 M, 30 ml). The organic phase was evaporated to a dark brown oil and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 40% EtOAc in heptane to give (6S, 8R) -6- (5-bromopyridin-2-yl) - 8-methyl-3- (tetrahydro-2H-pyran-2-yl) -7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline ( 0.142 g, 44%).

Mass / charge: ES ⁺ [M + n] ⁺ = 509/511.

- 87 037533

Alternative method # 1 for example 17.

Preparation of N- [1- (3-fluoropropyl) azetidin-3-yl] -6-G (6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9tetrahydropyrazolo [4,3-f] isoquinolin-6-yl1pyridin-3-amine

Trifluoroacetic acid (1.30 L, 17.04 mol) was added to a stirred solution of tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate (0.50 kg, 2.04 mol) in DCM (1.50 l) at 0 ° C for 0.5 h. The mixture was heated to 20 ° C and stirred at 20 ° C for 2.5 h. The mixture was heated to 30 ° C and stirring was continued for 1 h. The mixture was concentrated at reduced pressure and the residue was concentrated under reduced pressure from toluene (3x3.00 L). The resulting residue was dissolved in 1,4-dioxane (3.00 L) and treated with a 30% solution of sodium 2-methylbutan-2-olate in Me-THF (5.50 L, 13.63 mol) over 45 minutes, ensuring that the temperature does not exceed 29 ° C. A solution of 90% w / w (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H was added -pyrazolo [4,3-f] isoquinoline (0.81 kg, 1.70 mol) in 1,4-dioxane (1.50 L) and BrettPhos Pd G3 (0.077 kg, 0.085 mol) and the atmosphere was replaced with nitrogen using cycles of vacuum and nitrogen (3x). The mixture was heated to 55 ° C and stirred at 55 ° C for 2.5 h. The mixture was cooled to 21 ° C and an 8% solution of NaHCO ₃ in water (7.16 L, 6.82 mol) was added followed by the addition of isopropyl acetate (4, 50 l). The mixture was stirred for 2-3 minutes and the layers were separated. The organic layer was washed with 8% NaHCO ₃ in water (7.16 L, 6.82 mol), the mixture was stirred for 5 min and then the layers were separated. The combined aqueous layers were extracted with isopropyl acetate (4.50 L). The combined organic layers were treated with Silicycle (SiliaMetS-Thiol, 400 g, 1.26 mmol / g, 6 eq. With respect to Pd) and the mixture was stirred for 16 hours. The solids were removed by filtration through celite while washing the filter cake with isopropyl acetate. The filtrate was concentrated under reduced pressure (bath temperature 40 ° C). The residue was dissolved in isopropyl acetate (5.00 L) and treated with additional Silicycle (SiliaMetS-Thiol, 250 g, 1.26 mmol / g, 3.7 eq. Based on Pd). The resulting mixture was stirred at 21 ° C for 16 hours. The solids were removed by filtration through celite, rinsing the filter cake with isopropyl acetate (2.50 L). The filtrate was concentrated under reduced pressure (bath temperature 40 ° C) to give a brown foam. The crude product was purified by preparative SFC (Lux C4 column, 50 mm in diameter, 250 mm in length) using a gradient elution with EtOH / DEA 100 / 0.5 in CO ₂ , mobile phase at 140 bar at a flow rate of 400 ml / min at 40 ° C) to obtain a solid, which is a light brown foam. EtOAc (2.00 L) was added and the solution was concentrated under reduced pressure. EtOAc (1.90 L) and heptane (1.90 L) were added and the mixture was seeded at 25 ° C. The mixture was maintained at 25 ° C for 1 h. To the stirred mixture was added heptane (6.50 L) over 2 h. The resulting mixture was stirred at 22 ° C for 18 h. The resulting solid was collected by filtration while washing the filter cake with EtOAc / heptane 1: 5 (2.50 L). The solid was sucked off under nitrogen for 30 min, then dried under vacuum at 40 ° C for 9 days to give N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) - 8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine (626 g, 77 %) as an off-white solid.

¹ H NMR (600 MHz, DMSO-d6, 27 ° C) 1.07 (3H d) 1.63 (2H, dquin), 2.45 (2H, t), 2.72 (2H, br t), 2.83 (1H, br dd), 2.91-3.00 (1H, m), 3.06 (1H, br dd) 3.42-3.55 (2H, m), 3.58-3 , 65 (2H, m), 3.92 (1H, dquin) 4.43 (2H, dt) 4.92 (1H, s), 6.22 (1H, d), 6.79 (1H, d) 6.82 (1H, dd), 6.96 (1H, d), 7.21 (1H, d), 7.73 (1H, d), 8.04 (1H, s), 12.96 (1H , s);

mass / charge: ES + [M + H] + = 477.

Procedures used to prepare starting materials tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate and (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-7 - (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline are described below.

Preparation of tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate

The reaction was carried out in duplicate and the resultant was combined for processing. 1-Fluoro-3-iodopropane (189 g, 1.00 mol) was added to a stirred mixture of tert-butyl-N- (azetidine-3yl) carbamate; hydrochloride (200 g, 0.96 mol) and K ₂ CO ₃ (331 g, 2.40 mol) in THF (1.40 L) and water

- 88 037533 (0.50 mol) at 20 ° C. The reaction mixture was heated to 70 ° C and stirred at 70 ° C for 1.5 h. The two reaction mixtures were combined and added to water (2.5 L) and EtOAc (2.0 L). The mixture was stirred at 21 ° C for 3 minutes. The layers were separated and the organic layer was washed with water (2x2.0 L) and saturated ammonium chloride solution (2.0 L). The organic layer was concentrated under reduced pressure to obtain a crude product. Toluene (400 ml) was added and the mixture was heated to 60 ° C. Heptane (1.6 L) was added and the mixture was allowed to cool to 21 ° C and stirred for 15 minutes. The solid was collected by filtration and aspirated to give tert-butyl (1- (3fluoropropyl) azetidin-3-yl) carbamate (230 g, 52%) as a white solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.44 (9H, s), 1.68-1.80 (2H, m), 2.56 (2H, t), 2.87 (2H, s), 3.66 (2H, t), 4.29 (1H, s), 4.43 (1H, t), 4.52 (1H, t), 4.86 (1H, s).

Obtaining 4-bromo-1-tetrahydropyran-2-ylindazole

To a suspension of 4-bromo-III-indazole (1.50 kg, 7.60 mol) and 3,4-dihydro-2H-pyran (0.96 kg, 11.40 mol) in DCM (2.40 L) was added 4-methylbenzenesulfonic acid hydrate (11.60 g, 0.06 mol) at 20 ° C. under nitrogen. The resulting slurry was stirred at 20 ° C-29 ° C (slight exothermic effect) for 110 minutes. The resulting solution was washed with saturated aqueous NaHCO ₃ solution (2.00 L) and the organic layer was evaporated under reduced pressure. Hot (70 ° C) heptane (2.50 L) was added and the mixture was evaporated under reduced pressure to give a crystalline solid. Purification was carried out by recrystallization from hot (70 ° C) heptane (7.00 L) while allowing the solution to slowly cool to 34 ° C. The resulting solid was collected by filtration and washed with cold heptane to give 4-bromo-1-tetrahydropyran-2ylindazole (1.87 kg, 87%) as a crystalline solid.

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.54-1.66 (2H, m), 1.67-1.85 (1H, m), 1.94-2.09 (2H , m), 2.31-2.45 (1H, m), 3.70-3.81 (1H, m), 3.83-3.97 (1H, m), 5.88 (1H, dd ), 7.32-7.39 (1H, m), 7.40-7.44 (1H, m), 7.79 (1H, dt), 8.09 (1H, d);

mass / charge: ES- [M-H] ^- 282.

Preparation of tert-butyl-N - [(1R) -1-methyl-2- (1-tetrahydropyran-2-ylindazol-4-yl) ethyl] carbamate

A solution of 4-bromo-1- (tetrahydro-2H-pyran-2-yl) -III-indazole (0.70 kg, 2.49 mol) in THF (3.50 L) was gradually cooled in steps of 10 ° C. , up to -70 ° C. Added 2.5 M butyllithium in hexanes (1.10 L, 2.74 mol) over 70 minutes, keeping the internal temperature below -60 ° C. The resulting mixture was stirred at -72 ° C for 1 h. 2.3 M hexyllithium in hexanes (0.054 L, 0.12 mol) was added followed by the addition of a solution of tert-butyl (R) -4-methyl-1,2,3 α-oxathiazolidine-3-carboxylate 2,2-dioxide (652 g, 2.74 mol) in THF (2.80 L) over 2 hours, ensuring that the internal temperature does not exceed -58 ° C. The resulting mixture was stirred at -60 ° C for 16 hours. The mixture was slowly warmed to -10 ° C over 1.5 hours. Water (2.10 L) was added carefully. The layers were separated, and the organic layer was washed with saturated brine (1.40 L), and the organic layer was concentrated under reduced pressure to give an oil. IPA (1.50 L) was added and the mixture was concentrated under reduced pressure to give tert-butyl ((2R) -1- (1- (tetrahydro-2H-pyran-2yl) -1H-indazol-4-yl) propan-2 -yl) carbamate (1.27 kg,> 100%) as a brown oil. The resulting product was taken without additional purification.

1H NMR (500 MHz, CDCl3, 27 ° C) 1.08 (3H, dd), 1.43 (9H, s), 1.63-1.69 (1H, m), 1.73-1.82 (2H, m), 2.07 (1H, dd), 2.17 (1H, dd), 2.53-2.64 (1H, m), 2.95 (1H, s), 3.21 ( 1H, dt), 3.70-3.80 (1H, m), 3.97-4.12 (2H, m), 4.33-4.49 (1H, m), 5.71 (1H, ddd), 6.96 (1H, dd), 7.31 (1H, ddd), 7.46 (1H, d), 8.13 (1H, s);

mass / charge: ES ⁺ [M +] 359

Obtaining (2R) -1- (1 H-indazol-4-yl) propan-2-amine dihydrochloride

12 M HCl (1.00 L, 12.0 mol) was added over 20 min to a stirred solution of tert-butyl ((2R) -1- (1- (tetrahydro-2H-pyran-2-yl) -1H-indazol-4 -yl) propan-2-yl) carbamate (1.27 kg, 1.76 mol) in IPA (1.50 L), keeping the internal temperature below 30 ° C. The resulting mixture was heated to 40 ° C and stirred at 40 ° C for 16 h. The temperature was raised to 55 ° C and an additional portion of 12 M HCl (0.20 L, 2.40 mol) was added and stirring was continued for 7 h MTBE (2.50 L) was added and the mixture was allowed to stir at 25 ° C for 16 h. The resulting solid was collected by filtration and the filter cake was washed with 10% IPA in

- 89 037533

MTBE (2.00 l). The resulting solid was dried under reduced pressure at 40 ° C for 72 hours to obtain (2R) -1- (1H-indazol-4-yl) propan-2-amine; dihydrochloride (0.39 kg, 89%) as an off-white solid.

1H NMR (500 MHz, DMSO, 27 ° C) 1.12 (3H, d), 2.96 (1H, dd), 3.39 (1H, dd), 3.45- 3.55 (1H, m ), 6.94 (1H, d), 7.28 (1H, dd), 7.43 (1H, d), 8.23 (3H, s), 8.28 (1H, d);

mass / charge: eS ⁺ [M +] 175.

Preparation of (2R) -1- (1 H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine

A solution of 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.73 kg, 2.94 mol) in MeCN (2.00 L) was added to a stirred mixture of (R) -1- (1H-indazol-4-yl) propan-2- amine dihydrochloride (0.77 kg, 2.94 mol) and K ₂ CO ₃ (1.26 kg, 9.13 mol) in MeCN (8.00 L) at 25 ° C under nitrogen atmosphere. The resulting mixture was heated to 60 ° C and stirred at 60 ° C for 18 h. MeCN (5.00 L) was added and the solids were removed by filtration. The filtrate was concentrated under reduced pressure and the resulting solid was dissolved in EtOAc (10.00 L), washed with water (4.0 L) and diluted with saturated aqueous sodium chloride solution (4.8 L). The organic layer was concentrated under reduced pressure to give (2R) -1- (1H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (0.73 kg, 96%).

1H NMR (500 MHz, CDCl3, 27 ° C) 1.12 (3H, d), 2.96 (1H, dd), 3.08 (1H, dd), 3.14-3.33 (3H, m ), 6.98 (1H, d), 7.32 (1H, dd), 7.37-7.41 (1H, m), 8.13 (1H, d), 10.86 (1H, s) ;

mass / charge: ES ⁺ [M + H] ⁺ = 257.

Obtaining (6S, 8R) -6- (5-bromo-2-pyridyl) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9-tetrahydropyrazolo [4,3-f ] isoquinoline

Trifluoroacetic acid (0.66 L, 8.66 mol) was slowly added to a stirred mixture of 5-bromopicolinaldehyde (0.55 kg, 2.89 mol) and (R) -1- (1H-indazol-4-yl) -N - (2,2,2-trifluoroethyl) propane-2amine (0.83 kg, 2.89 mol) in toluene (7.30 L) at 21 ° C. The resulting solution was heated to 80 ° C and stirred at 80 ° C for 18 h. The temperature was raised to 90 ° C and heating was continued for 4 h. The mixture was cooled to 80 ° C and an additional portion of 5-bromopicolinaldehyde (0.015 kg, 0.081 mol) and the solution was stirred at 80 ° C for 16 hours. The mixture was cooled to 21 ° C and saturated aqueous NaHCO ₃ solution (6.60 L) was added over 20 minutes. EtOAc (5.00 L) was added and the layers were separated. The organic layer was washed with saturated aqueous NaHCO ₃ solution (3.30 L) and concentrated under reduced pressure. Ethanol was added and the mixture was concentrated under reduced pressure to give the crude product as a crumbly solid. cis-Isomer removed by preparative SFC (SuperSep 1 column (packed with CelluCoat ™ 10 μm), 50 mm in diameter, 250 mm in length), elution with 28% EtOH in CO ₂ , mobile phase at 140 bar at 450 ml / min flow rate at 30 ° C) to give (6S, 8R) -6- (5-bromo-2-pyridyl) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9 α-tetrahydropyrazolo [4,3-f] isoquinoline (0.94 kg, 77%) as a pale yellow solid.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.16 (3H, d), 2.89 (1H, dd), 2.99 (1H, dq), 3.25-3.34 (2H , m), 3.55 (1H, td), 5.10 (1H, s), 6.93 (1H, d), 7.22 (1H, d), 7.40 (1H, d), 7 , 75 (1H, dd), 8.05 (1H, d), 8.56 (1H, dd), 10.24 (1H, s);

mass / charge: ES + [M + H] + = 427.

Alternative Method # 2 for Example 17.

Receiving

N- (3-fluoropropyl) azetidin-3-yl] -6 - [(6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9 tetrahydropyrazolo [4, 3- £] isoquinolin-6-yl1pyridin-3-amine

1- (3-Fluoropropyl) azetidin-3-amine (6.31 g,

37.98 mmol) was added to a solution of (6S, 8R) -6- (5- 90 037533 bromopyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (15.47 g, 34.92 mmol) in 1,4-dioxane (175 ml). The resulting solution was degassed under vacuum for 5 min, then filled with nitrogen (x2). Sodium tert-butoxide (13.43 g, 139.69 mmol) was added followed by the 3rd generation BrettPhos precatalyst (0.95 g, 1.05 mmol). The resulting mixture was heated at 55 ° C for 18 h. The mixture was poured into EtOAc and water. Brine was added and the layers were separated. The organic layer was washed with saturated aqueous sodium chloride solution (x3) and the combined aqueous layers were extracted with EtOAc.

The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 50% methanol in ethyl acetate. Fractions containing the desired product were combined, concentrated under reduced pressure and then re-purified by flash chromatography on silica, elution gradient 0 to 40% EtOAc in hexanes, to give a red-orange solid. This solid was dissolved in 10% MeOH in DCM, filtered, and the filter cake washed with DCM. The combined filtrates were concentrated under reduced pressure to give N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine (15.28 g, 92%) as a light orange foam solid. trans-cis-Isomers were separated by preparative SFC ((S, S) Whelk-O1 column, 30 mm diameter, 250 mm long) eluting with 30% (0.2% NH4OH in MeOH) in CO ₂ , mobile phase at 100 bar at a flow rate of 20 ml / min at 40 ° C). Fractions containing the trans isomer were concentrated under reduced pressure. The resulting residue was further purified by flash chromatography on silica, elution gradient from 0 to 30% MeOH in EtOAc. The fractions containing the desired product were concentrated under reduced pressure, then concentrated from MeCN (x2) to give N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl- 7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (10.76 g, 74%) as a pale yellow foam / solid.

1H NMR (600 MHz, DMSO-d ₆ , 27 ° C) 1.07 (3H d) 1.63 (2H, dquin), 2.45 (2H, t), 2.72 (2H, br t), 2.83 (1H, br dd), 2.91-3.00 (1H, m), 3.06 (1H, br dd) 3.42-3.55 (2H, m), 3.58-3 , 65 (2H, m), 3.92 (1H, dquin) 4.43 (2H, dt) 4.92 (1H, s), 6.22 (1H, d), 6.79 (1H, d) 6.82 (1H, dd), 6.96 (1H, d), 7.21 (1H, d), 7.73 (1H, d), 8.04 (1H, s), 12.96 (1H , s);

mass / charge: ES + (M + H) +477.

The SFC fractions containing the cis isomer were concentrated under reduced pressure and the resulting residue was further purified by flash chromatography on silica, elution gradient from 0 to 30% MeOH in EtOAc. The fractions containing the desired product were concentrated under reduced pressure, and the resulting residue was filtered to give N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6R, 8R) -8-methyl-7- (2 , 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (1.13 g, 8%) as a solid , which is a light yellow foam.

1H NMR (300 MHz, DMSO-d6, 27 ° C) 1.28 (3H, d), 1.55-1.75 (2H, m), 2.42-2.49 (2H, m), 2 , 76 (2H, br t), 2.80-2.92 (1H, m), 3.07-3.18 (1H, m), 3.18-3.28 (1H, m), 3, 38-3.54 (2H, m), 3.59-3.70 (2H, m), 3.95 (1H, sxt), 4.45 (2H, dt), 5.11 (1H, s) , 6.21 (1H, d), 6.72 (1H, d), 6.82 (1H, dd), 7.02 (1H, d), 7.19 (1H, d), 7.78 ( 1H, d), 8.06 (1H, s), 12.94 (1H, s);

mass / charge: ES ⁺ (M + H) +477.

Procedures used to obtain starting material, which is (6S, 8R) -6 (5-bromopyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinoline are described below.

Preparation of 2,2,2-trifluoro-N - [(1R) -2- (1H-indazol-4-yl) -1-methylethyl] acetamide

TEA (26.0 ml, 186.5 mmol) was added via a syringe to a stirred slurry of (R) -1- (1Hindazol-4-yl) propan-2-amine dihydrochloride (15.29 g, 61.61 mmol) and ethyl 2,2,2-trifluoroacetate (8.09 ml, 67.8 mmol) in MeOH (150 ml) at room temperature. The resulting solution was stirred at room temperature for 18 hours, then concentrated under reduced pressure. The resulting solid was then dissolved in DCM (40 ml) and the residual triethylamine hydrochloride was precipitated by the addition of Et ₂ O (300 ml). The slurry was stirred for 5 minutes, then filtered, and the filtrate was concentrated under reduced pressure to give a solid. The solid was dissolved in EtOAc and saturated aqueous sodium chloride solution. The layers were separated and the organic layer was washed with saturated saturated aqueous sodium chloride solution, dried over MgSO ₄ , filtered and concentrated to give (R) -N- (1- (1H-indazol-4-yl) propan-2-yl) -2, 2,2trifluoroacetamide (16.51 g, 99%) as an off-white solid.

- 91 037533

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.19 (3H, d), 3.03 (1H, dd), 3.13 (1H, dd), 4.13-4.30 ( 1H, m), 6.89 (1H, d), 7.25 (1H, dd), 7.38 (1H, d), 8.18 (1H, t), 9.35 (1H, br d) , 13.01 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ 272.

Preparation of (2R) -1- (1H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine

1 M borane-THF complex in THF (365 ml, 365.0 mmol) was rapidly added dropwise via cannula to a stirred solution of (R) -N- (1- (1H-indazol-4-yl) propan-2-yl) -2,2,2-trifluoroacetamide (16.5 g, 60.8 mmol) in THF (100 mL) at room temperature (gas evolution; slight exotherm observed during addition). The reaction mixture was then heated at 60 ° C for 15.5 h, then allowed to cool to room temperature. The reaction mixture was placed in a water bath at ambient temperature and MeOH (80 ml) was slowly added dropwise. The reaction mixture was concentrated under reduced pressure, and the resulting residue was dissolved in MeOH (80 ml) and concentrated under reduced pressure. The resulting residue was dissolved in MeOH (250 ml) and placed in a water bath at ambient temperature. A slurry of 10% palladium-carbon (6.47 g, 6.08 mmol) in MeOH (90 ml) was added, rinsing the flask with MeOH (10 ml) (slight evolution of gas was noted). The mixture was stirred at room temperature for approximately 3 minutes, then heated at 65 ° C for 2 hours. After cooling to room temperature, the mixture was filtered through celite and concentrated under reduced pressure. The pale yellow residue was dissolved in DCM and filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient 10% to 60% EtOAc in hexanes. The fractions containing the desired product were concentrated under reduced pressure to give (R) -1- (1H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (14.21 g, 86%) as a pale yellow oil.

1H NMR (300 MHz, DMSO-d6, 27 ° C) 0.91 (3H, d), 2.17-2.30 (1H, m), 2.71 (1H, dd), 2.98-3 , 17 (2H, m), 3.22-3.38 (2H, m), 6.89 (1H, d), 7.24 (1H, dd), 7.35 (1H, d), 8, 12 (1H, t), 12.97 (1H, br s);

mass / charge: ES ⁺ [M + H] ⁺ 258.

Obtaining (6S, 8R) -6- (5-bromo-2-pyridyl) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9-tetrahydropyrazolo [4,3-f ] isoquinoline

Br

5-Bromopicolinaldehyde (9.80 g, 52.7 mmol) was added to a stirred solution of (R) -1- (1Hindazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (14 , 05 g, 51.66 mmol) in toluene (246 mL) under nitrogen. Then added trifluoroacetic acid (12.30 ml) and the reaction mixture was heated at 90 ° C for 4.5 h. Allowed to cool the mixture to room temperature and added EtOAc (200 ml). The mixture was cooled to 5 ° C and the mixture was podslushivaet by the slow addition of saturated aqueous NaHCO ₃ solution. The resulting layers were separated and the aqueous layer was extracted with EtOAc (2x80 ml). The combined organic extracts were washed with saturated aqueous NaHCO ₃ solution, saturated aqueous sodium chloride solution, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 5% to 60% EtOAc in hexanes. The fractions containing the desired product were concentrated under reduced pressure, then dried in vacuo at 50 ° C for 3 h to give (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-7- ( 2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (19.29 g, 88%) as a pale orange solid. 1H NMR analysis revealed 10: 1 trans / cis isomers.

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.10 (3H, d), 2.83-2.94 (1H, m), 2.95-3.14 (2H, m), 3.34-3.47 (1H, m), 3.49-3.72 (1H, m), 5.10 (1H, s), 6.88 (1H, d), 7.25 (1H, d), 7.33 (1H, d), 7.99 (1H, dd), 8.06 (1H, d), 8.56 (1H, d), 13.00 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ 425.

Alternative Methods for Intermediates Used in the Synthesis of Example 17.

Preparation of N - [(1R) -2- (3-amino-2-methylphenyl) -1-methylethyl] -2,2,2-trifluoroacetamide

A solution of (R) -3- (2-aminopropyl) -2-methylaniline (0.51 g, 3.13 mmol), DIPEA (0.55 ml, 3.13 mmol) and

- 92 037533 ethyl 2,2,2-trifluoroacetate (0.47 ml, 3.91 mmol) in MeOH (9.5 ml) was stirred at room temperature under nitrogen atmosphere for 16 hours. The reaction mixture was concentrated under reduced pressure. DCM and water were added and the layers were separated. The aqueous layer was extracted with DCM (3x50 ml) and the combined organic layers were dried (Na ₂ SO ₄ ) and filtered. The crude product was purified by flash chromatography on silica, elution gradient 0 to 100% EtOAc in hexane. The pure fractions were evaporated to dryness to give N - [(1R) -2- (3-amino-2methylphenyl) -1-methylethyl] -2,2,2-trifluoroacetamide (680 mg, 83%).

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.06-1.14 (m, 3H), 2.01 (s, 3H), 2.54-2.71 (m, 1H), 2.72-2.84 (m, 1H), 4.04-4.12 (m, 1H), 4.60-4.77 (m, 2H), 6.24-6.37 (m, 1H ), 6.43-6.52 (m, 1H), 6.72-6.84 (m, 1H), 9.19-9.32 (m, 1H);

mass / charge: ES ⁺ [M + H] ⁺ = 261.

Preparation of 2-methyl-3 - [(2R) -2- (2,2,2-trifluoroethylamino) propyl] aniline

1.0 M borane in THF, in THF (19.4 mL, 19.37 mmol) was added to a stirred solution of (R) -N- (1 (3-amino-2-methylphenyl) propan-2-yl) -2 , 2,2-trifluoroacetamide (840 mg, 3.23 mmol) in THF (10 ml). The reaction mixture was heated at 65 ° C under nitrogen for 6 h. The mixture was allowed to cool to room temperature and MeOH was added carefully. The resulting mixture was stirred at room temperature for 30 minutes and then concentrated under reduced pressure. The resulting residue was dissolved in MeOH and heated to 65 ° C for 6 hours. The mixture was allowed to cool to room temperature and then concentrated under reduced pressure to give the crude product, which was purified by flash chromatography on silica, elution gradient from 0 to 100% EtOAc in hexane. The product fractions were evaporated to dryness to give 2-methyl-3 - [(2R) -2- (2,2,2-trifluoroethylamino) propyl] aniline (665 mg, 84%).

1H NMR (300 MHz, DMSO-d6, 27 ° C) 0.89 (s, 3H), 1.98 (s, 3H), 2.08-2.22 (m, 1H), 2.25-2 , 42 (m, 1H), 2.69-2.87 (m, 2H), 3.15-3.29 (m, 2H), 4.61-4.75 (m, 2H), 6, 30-6.40 (m, 1H), 6.43-6.54 (m, 1H), 6.71-6.83 (m, 1H);

mass / charge: ES ⁺ [M + H] ⁺ = 247.

Preparation of (2R) -1- (3-bromo-2-methylphenyl) -N- (2,2,2-trifluoroethyl) propan-2-amine

DIPEA (2.99 ml, 17.10 mmol) was added to (R) -1- (3-bromo-2-methylphenyl) propan-2-amine (1.30 g, 5.70 mmol) and 2.2, 2-trifluoroethyl trifluoromethanesulfonate (0.23 M in DCM, 29.7 ml, 6.84 mmol) in 1,4-dioxane (25 ml) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 85 ° C for 16 hours. The reaction mixture was concentrated under reduced pressure and the crude product was purified by flash chromatography on silica, elution gradient from 0 to 50% EtOAc in heptane. The fractions containing the product were concentrated under reduced pressure to give (R) -1- (3-bromo-2-methylphenyl) -N- (2,2,2-trifluoroethyl) propan-2-amine (0.56 g, 32 %) in the form of oil.

¹ H NMR (500 MHz, DMSO, 27 ° C) 0.91 (3H, d), 2.26 (1H, q), 2.34 (3H, s), 2.78-2.86 (1H, m), 2.89 (1H, dd), 3.19-3.28 (2H, m), 7.01-7.06 (1H, m), 7.15 (1H, dd), 7.43 (1H, dd);

mass / charge: ES + [M + H] ⁺ = 310/312.

Preparation of 2-methyl-3 - [(2R) -2- (2,2,2-trifluoroethylamino) propyl] aniline

tris- (Dibenzylideneacetone) dipalladium (0) (0.050 g, 0.05 mmol) and (±) -2,2'-bis- (diphenylphosphino) 1,1'-binaphthalene (0.067 g, 0.11 mmol) were added to suspensions of (R) -1- (3-bromo-2-methylphenyl) -N- (2,2,2trifluoroethyl) propan-2-amine (0.56 g, 1.81 mmol), benzophenone imine (0.33 ml, 1.99 mmol) and sodium tert-butoxide (0.26 g, 2.71 mmol) in degassed toluene (7 ml) and the reaction mixture was heated to 90 ° C for 16 h. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was dissolved in DCM (50 ml) and washed with water (50 ml). The aqueous layer was extracted with DCM (25 ml) and the combined organics were concentrated to ~ 25 ml. 2M HCl solution (50 ml) was added and the biphasic mixture was stirred vigorously for 30 min. The layers were separated and the aqueous layer was washed with DCM. The aqueous phase was podslushivaet by adding 2 M aqueous NaOH solution. The resulting mixture was extracted with DCM (2x100 ml) and the combined organic extracts were concentrated under reduced pressure to give

- 93 037533 (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (0.31 g, 70%) as an oil.

¹ H NMR (500 MHz, DMSO, 27 ° C) 0.90 (3H, d), 1.98 (3H, s), 2.12-2.18 (1H, m), 2.30-2, 36 (1H, m), 2.742.82 (2H, m), 3.17-3.29 (2H, m), 4.69 (2H, s), 6.35 (1H, dd), 6.48 (1H, dd) 6.78 (1H, t);

ES ⁺ [M + H] ⁺ = 247.

Preparation of (1S, 3R) -1- (5-bromo-2-pyridyl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -3,4-dihydro-1Nisoquinolin-6-amine

The salt of trifluoromethanesulfonic acid and ytterbium (III) (0.17 g, 0.27 mmol) was added to a stirred solution of (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (1.35 g, 5.49 mmol), 5-bromopicolinaldehyde (1.02 g, 5.49 mmol) and water (0.49 ml, 27.43 mmol) in MeCN (21.5 ml). The resulting solution was stirred at 80 ° C for 1 h. The mixture was concentrated under reduced pressure and the residue was loaded dry onto silica gel and purified by flash chromatography on silica, elution gradient from 10% to 30% EtOAc in hexane. The fractions containing the desired product were concentrated to dryness to give (1S, 3R) -1- (5-bromopyridin-2-yl) -3,5-dimethyl2- (2,2,2-trifluoroethyl) -1.2, 3,4-tetrahydroisoquinoline-6-amine (2.22 g, 98%) as a yellowish gum.

¹ H NMR (300 MHz, DMSO-d6, 27 ° C) 0.88 (3H, d), 1.91 (3H, s), 2.37-2.46 (1H, m), 2.60- 2.76 (1H, m), 2.76-3.00 (1H, m), 3.15-3.28 (1H, m), 3.34-3.63 (1H, m), 4, 65 (2H, s), 4.85 (1H, s), 6.19-6.52 (2H, m), 7.107.31 (1H, m), 7.97 (1H, dd), 8.62 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 414.

Obtaining (6S, 8R) -6- (5-bromo-2-pyridyl) -8-methyl-7- (2,2,2-trifluoroethyl) -3,6,8,9-tetrahydropyrazolo [4,3-f ] isoquinoline

Br

V = N

HN \ / N— \ F

VF (1S, 3R) -1- (5-Bromopyridin-2-yl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1,2,3,4-tetrahydroisoquinolin6-amine (2 , 21 g, 5.33 mmol) in propionic acid (14.0 ml) was cooled to -20 ° C. A solution of sodium nitrite (0.40 g, 5.86 mmol) in water (2.8 ml) was added dropwise over 5 minutes while maintaining the bath temperature between -20 and -15 ° C. The resulting mixture was stirred at -20 ° C for 45 minutes. Toluene (80 ml), cooled to -15 ° C, was poured into the reaction mixture and the reaction mixture was stirred for 15 min. A solution of sodium carbonate (12.0 g, 113 mmol) in water (125 ml) was added slowly and the mixture was allowed to warm to room temperature. Added EtOAc (100 ml) and the layers were separated. The aqueous layer was extracted with CHCl ₃ / IPA (3: 1) (100 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the crude product, which was purified by flash chromatography on silica, elution gradient 15% to 50% EtOAc in hexane. The fractions containing the product were evaporated to dryness to give (6S, 8R) -6- (5-bromopyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8 , 9tetrahydro-3H-pyrazolo [4.3- £] isoquinoline (1.63 g, 72%) as a brown solid.

Ή NMR (300 MHz, DMSO-d6, 27 ° С) 1.10 (3H, d), 2.83-2.96 (1H, m), 2.96-3.14 (2H, m), 3 , 35-3.48 (1H, m), 3.49-3.73 (1H, m), 5.11 (1H, s), 6.87 (1H, d), 7.27 (1H, d ), 7.35 (1H, d), 7.99 (1H, dd), 8.09 (1H, s), 8.538.63 (1H, m), 12.96 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 425.

Preparation of tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate P О pN ^^ F

Η

Suspension of tert-butylazetidin-3-ylcarbamate (10.00 g, 58.06 mmol), 1-fluoro-3-iodopropane (11.13 g, 59.22 mmol) and potassium carbonate (16.05 g, 116.13 mmol) in MeCN (200 ml) was stirred at room temperature for 2 days. The solids were removed by filtration and the filtrate was concentrated under reduced pressure. The residue was taken up in water and extracted with DCM. The combined organic extracts were dried over N ₂ aSO ₄ , filtered and concentrated under reduced pressure to give tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate (13.40 g, 99%) as a pale yellow solid matter.

Ή NMR (300 MHz, CDCl ₃ , 27 ° C) 1.45 (9H, s), 1.61-1.92 (2H, m), 2.51-2.67 (2H, m), 2, 85 (2H, br s), 3.57-3.72 (2H, m), 4.30 (1H, br d), 4.36-4.63 (2H, m), 4.89 (1H, brd).

- 94 037533

Alternative preparation of tert-butyl (1 - (3-fluoropropyl) azetidin-3-yl) carbamate

Added trifluoromethanesulfonic anhydride (2.31 ml, 13.75 mmol) followed by the addition of 2,6-dimethylpyridine (1.74 ml, 15.00 mmol) to a solution of 3-fluoropropan-1-ol (0.94 ml, 12 , 5 mmol) in DCM (47 ml) at 0 ° C and the reaction mixture was stirred at 0 ° C for 1 h. The reaction mixture was washed with 1N. HCl, then the organic layer was dried and carefully evaporated. The residue was then added to a separate vial containing a suspension of tert-butylazetidine-3yl carbamate (1.94 g, 11.25 mmol) and DIPEA (3.24 ml, 18.75 mmol) in dioxane / DCM (1: 1, 40 ml ) and the reaction mixture was stirred at room temperature (exothermic effect with the addition of triflate). The reaction mixture was diluted with DCM and washed with NH4Cl solution. The organic layer was dried and evaporated under reduced pressure to give the crude product. The crude product was purified by flash chromatography on silica, gradient elution 0 to 10% 1 M NHs / MeOH in DCM. Pure fractions were evaporated to dryness to give tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate (2.82 g, 97%) as a purple oil. ¹ H NMR as above.

Obtaining 1 - (3-fluoropropyl) azetidine-3-amine

Trifluoroacetic acid (39.8 ml, 517 mmol) was added dropwise over 30 min to a solution of tert-butyl (1- (3-fluoropropyl) azetidin-3-yl) carbamate (20.0 g, 86 mmol) in DCM ( 67.8 ml) which was immersed in a water bath. After 18 hours, the reaction mixture was concentrated under reduced pressure to an orange oil. This oil was dissolved in 10% methanol in DCM (125 ml) and potassium carbonate (71.4 g, 517 mmol) was added with vigorous stirring. After 15 minutes. added another 30 g of potassium carbonate. Stirring became slower and celite (~ 15 g) was added. Stirring was continued for another 15 minutes and the mixture was filtered, washing with 10% methanol in DCM. The filtrate was concentrated under reduced pressure on a rotary evaporator (pressure: 100 mbar, water bath temperature: 30 ° C) and the resulting orange oil was purified by flash chromatography on silica (detection λ = 210 nm), elution gradient 0 to 15% (2 M ammonia in methanol) in DCM to give 1- (3-fluoropropyl) azetidin-3-amine (12.8 g) contaminated with 28 wt% methanol, based on NMR integration, as a yellow oil. This material could be directly applied, stored in a freezer for later use, or further purified by vacuum distillation according to the following example. 1- (3-Fluoropropyl) azetidin-3-amine (14.8 g) as a light yellow oil was distilled using a molecular distillation apparatus under cooling conditions with a water jacket under vacuum (vacuum pump). The flask containing the amine was immersed in an oil bath and the temperature of the bath was gradually raised to 140 ° C over a period of 30 minutes. Distillation took place at a bath temperature of 110-135 ° C and an approximate top temperature of 70 ° C to give 1- (3-fluoropropyl) azetidine-3-amine (12.4 g, 84%) as a clear colorless oil. ... This oil was stored in the freezer until used.

1H NMR (300 MHz, DMSO-d ₆ , 27 ° С) 1.52-1.70 (2H, m), 1.75 (2H, br s), 2.38 (2H, t), 2.43 -2.48 (2H, m), 3.27-3.39 (1H, m), 3.42-3.48 (2H, m), 4.43 (2H, dt);

mass / charge: ES + [M + H] + = 133.

Example 17A. Preparation of Form A (anhydrous form) N- (1- (3-fluoro-propyl) azetidin-3-yl) -6 (6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine

Method 1.

4.0 mg amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6.7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was dissolved in 100 μl EtOAc to give a clear solution. Approximately 200 μl of heptane was slowly added to the solution and a cloudy suspension formed. The slurry was stirred at ambient conditions for 3 days. Form A crystalline material was obtained.

Method 2.

1.60 g of amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6.7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was suspended in 15 ml of acetone / H2O (10: 1). A slurry formed and then a gel formed. 10 mg form A N- (1- (3-fluoropronyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7, 8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was seeded after 2 hours and the suspension was stirred at ambient temperature for 1 day while the gel turned into solid precipitation. The solid sediments on the wall and bottom were manually transferred into the solution and a homogeneous suspension was obtained after stirring for 1 hour. The solid was collected by filtration.

- 95 037533 and washed twice with 5.0 ml of a mixture of acetone / H 2 O (1:10), followed by several rinsing with H 2 O. 1.45 g of a white powder (90% yield) was obtained as Form A after drying in vacuo for 4 h.

Method 3.

501 mg of amorphous material representing N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine was dissolved in 1.0 ml EtOAc to give a brown solution. 1.0 ml of heptane was slowly added to the solution. At the end of the addition, a solid began to form, but then dissolved. 2-5 mg form A N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was added to the clear brown solution and a solid began to precipitate. The suspension was stirred at ambient temperature for 1 day. The solid was collected by filtration and dried under air. Received 345 mg of an off-white powder (yield: ~ 69%) as Form A.

Form A from Method 2 was analyzed by XRPD and the results are shown in Table II. 1 and shown in FIG. one.

Table 1

XRPD peaks for Form A

Angle (2ftt0.2 °) Intensity (%) 21.1 100.0 20.8 54.3 14.6 41.9 18.6 41.6 12.3 38.9 15.5 34.1 24.6 31.3 15.8 30.6 13.4 23.2 19.0 21,7

Form A was analyzed using thermal analysis techniques. DSC analysis indicated that Form A had a melting point with an onset at 132 ° C and a peak at 137 ° C. TGA results indicated that Form A had a weight loss of about 0.2% when heated from about 25 ° C to about 100 ° C. An exemplary Form A DSC / TGA thermogram is shown in FIG. 2.

Example 17B. Preparation of Form B N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (TBME solvate).

100 mg N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8, 9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6-ul) pyridin-3-amine (amorphous material) was partially dissolved in 1.0 ml of TBME. The slurry was stirred at ambient conditions whereby additional solid precipitated in the slurry. The suspension was stirred for 2 h.

Form B N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8, 9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6-yl) pyridin-3-amine was identified when the slurry was directly weighed into a sample holder for XRPD characterization. Form B was converted to Form C after the solid was filtered and dried at ambient conditions.

Form B was analyzed by XRPD, and the results are shown in table. 2 and shown in FIG. 3.

table 2

XRPD peaks for Form B

Angle (20 ± O, 2 °) Intensity (%) 6.0 100.0 17.8 67.0 18.4 34.9 16.3 30.3 21.5 29.2 12.3 27.0 17.2 24.8 13.4 23.3 18.1 21.6 15.6 20.6

- 96 037533

Example 17C. Preparation of form C N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (solvate with

TVME).

200 mg of amorphous material representing N- (1- (3-fluoropropyl) azetugin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyrugin3-amine was almost completely dissolved in 1 ml of TBME. The suspension was stirred at ambient conditions whereby additional solid precipitated after 10 minutes. The slurry was stirred for 2 hours and then evaporated at ambient conditions. Form C was obtained after the resulting solid was dried under ambient conditions.

Form C was analyzed by XRPD, and the results are shown in table. 3 and shown in FIG. four.

Table 3

XRPD peaks for Form C

Angle (20 ± O, 2 °) Intensity (%) 6.8 100.0 18.6 64.1 13.9 62.5 18.0 61.6 17.2 53.4 17.4 52.4 20.5 47.3 19.2 40.0 20,7 37.4 13.7 36.3

Form C was analyzed using thermal analysis techniques. The results of DSC analysis indicated that Form C was characterized by an endothermic desolvation phenomenon with an onset at 75 ° C and a peak at 84 ° C. TGA results indicated that Form C had a weight loss of about 7.1% when heated from about 25 ° C to about 150 ° C. An exemplary DSC / TGA Form C thermogram is shown in FIG. five.

Example 17D. Preparation of Form D N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9 -tetragygro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyrigin-3-amine (solvate with CPME).

150 mg of amorphous material representing N- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine was stirred in 2.0 ml of CPME (cyclopentyl methyl ether). After stirring for 1 day, the solid was isolated by filtration and dried under air. Received ~ 50 mg of a white powder, which is form D.

Form D was analyzed by XRPD, and the results are shown in table. 4 and shown in FIG. 6.

Table 4

XRPD peaks for Form D

Angle (20 ± O, 2 °) Intensity (%) 6.2 100.0 18.3 55.0 18.9 24.3 16.1 21.8 16.8 18.4 22.2 17.9 14.1 17.1 19.9 13.5 22.5 13.5 23.9 13.4

Form D was analyzed using thermal analysis techniques. The results of DSC analysis indicated that Form D was characterized by an endothermic desolvation phenomenon with an onset at 76 ° C and a peak at 81 ° C followed by another endothermic phenomenon with an onset at 128 ° C and a peak at 133 ° C. TGA results indicated that Form D had a weight loss of about 6.8% when heated from about 25 ° C to about 150 ° C. An exemplary DSC / TGA form D thermogram is shown in FIG. 7.

- 97 037533

Example 17E. Preparation of Form E (anhydrous form) H- (1- (3-fluoropropyl) azetidin-3-yl) -6 ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin3-amine

Method 1.

100 mg amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was added to 2.0 ml of heptane to form a slurry. Added 0.40 ml EtOAc. After stirring for 2 h, an additional 0.10 mL of EtOAc was added to the slurry and the slurry was stirred at ambient conditions for 18 h, while a new crystalline form, Form E, was identified by XRPD. 75 mg of a white solid form E was obtained after the solid was isolated and dried at ambient conditions (yield: 75%).

Method 2.

1.002 g of amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was dissolved in 5.0 ml EtOAc to give a clear light brown solution. 5.0 ml of heptane was added and the solution remained clear. Added 10 mg of Form E N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2.2.2trifluoroethyl) -6.7.8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine as a seed and the suspension was stirred at ambient temperature. A solid began to precipitate at ambient conditions and a wet precipitate formed after 1 h. 5.0 ml EtOAc was slowly added to the slurry and the resulting slurry was stirred at ambient temperature for 1 h. Another 10 ml heptane was added, more solid began to precipitate ... After stirring at ambient conditions for 2 h, the slurry was heated to 60 ° C and stirred at 60 ° C for 2 h. The slurry was cooled to ambient temperature and stirred for 18 h. Off-white solid, Form E , was isolated by filtration and washed 3 times with a mixed solvent EtOAc / heptane 1: 4. 0.875 g of a pale white powder was collected after drying under air (yield: 87%).

Form E from Method 1 was analyzed by XRPD and the results are shown in Table II. 5 and shown in FIG. eight.

Table 5

XRPD peaks for Form E

Angle (20 ± O, 2 °) Intensity (%) 17.9 100.0 14.8 67.1 20.9 60.1 23.1 55.4 16.2 49.3 20.0 35.6 18.2 32.9 12.3 30.4 22.2 19.0 13.9 18.9

Form E was analyzed using thermal analysis techniques. DSC analysis indicated that Form E had a melting point with an onset at 126 ° C and a peak at 133 ° C. TGA results indicated that Form E had a weight loss of about 0.8% when heated from about 25 ° C to about 100 ° C. An exemplary Form E DSC / TGA thermogram is shown in FIG. nine.

Example 17F. Preparation of Form F N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (heptane solvate).

mg amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was dissolved in 200 μl of acetone. 1.0 ml of heptane was slowly added to the solution. The solution started to turn cloudy and then a slurry formed. The slurry was stirred at ambient temperature after adding 1.0 ml of heptane. Needle-shaped particles were observed in suspension. The slurry was heated to 60 ° C and stirred for 2 hours and then cooled to ambient temperature. Needle-like particles were formed.

The resulting Form F slurry was analyzed by XRPD and the results are shown in FIG. 10. Form F crystallinity decreased during XRPD characterization and a second XRPD cycle indicated amorphous material after drying in the sample holder.

- 98 037533

Example 17F. Preparation of Form G N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9 -tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (methylpentanone solvate).

100 mg amorphous N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine was suspended in 200 μl of methylpentanone and 2-5 mg of form A N- (1- (3-fluoropropyl) azetidine- 3-yl) -6 - ((6S, 8R) -8-methyl-7 (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline -6-yl) pyridin-3-amine, a precipitate formed after stirring at ambient conditions for 10 minutes. Added 200 μl of methylpentanone, and a suspension formed. The slurry was stirred at ambient conditions for 18 hours. XRPD results indicated that a crystalline form, Form G N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8- methyl 7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine. Form G was converted to a partially amorphous material after isolation and drying at ambient conditions.

Form G was analyzed by XRPD, and the results are shown in table. 6 and shown in FIG. 10.

Table 6

XRPD peaks for Form G

Angle (2ftt0.2 °) Intensity (%) 6.2 100.0 18.1 95.9 16.6 38.8 18.7 37.5 21.9 36.5 15.9 33.9 22.4 28.1 19.7 26.4 14.0 24.6 12.6 23.9

Example 18.

5-Fluoro-6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4,3

f] isoquinolin-6-yl) -N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

[(2-Dicyclohexylphosphino-3,6-dimethoxy-2 ', 4', 6'-triisopropyl-1,1'-biphenyl) -2- (2'-amino-1,1'biphenyl)] palladium (P) methanesulfonate (Brett Phos G3) (48.6 mg, 0.05 mmol) was added to a degassed solution of 1- (3-fluoropropyl) azetidine-3-amine (95 mg, 0.72 mmol), (6S, 8R) -6 - (5-bromo-3-fluoropyridine-2yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline ( 155 mg, 0.36 mmol) and sodium tert-butoxide (206 mg, 2.15 mmol) in 1,4-dioxane (3 ml) under nitrogen. The resulting solution was stirred at 90 ° C for 2 h. The reaction mixture was diluted with EtOAc (50 ml) and washed sequentially with water (50 ml) and saturated aqueous sodium chloride solution (50 ml). The organic layer was dried with MgSO ₄ , filtered and evaporated to give the crude product, which was purified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5 μm silica, 50 mm in diameter, 100 mm in length) using water mixtures ( containing 0.1% NH ₃ ) and MeCN with decreasing polarity as eluents. The fractions containing the desired compound were evaporated to dryness to give 5-fluoro-6 ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H -pyrazolo [4,3-f] isoquinolin-6-yl) N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine (39.0 mg, 22%) as a white solid ...

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.48-0.61 (2H, m), 0.92-1.09 (2H, m), 1.13 (3H, d), 1, 68-1.83 (2H, m), 2.56-2.62 (2H, m), 2.67 (1H, dd), 2.86 (1H, dd), 2.90-2.95 ( 2H, m), 3.17 (1H, dd), 3.27 (1H, dd), 3.70 (2H, q), 3.87 (1H, td), 4.04 (1H, q), 4.21 (1H, d), 4.43 (1H, t), 4.53 (1H, t), 5.39 (1H, s), 6.50 (1H, dd), 6.79 (1H , d), 7.12 (1H, d), 7.68 (1H, d), 8.03 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 485.

5-Fluoro-6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline-6- yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine was prepared as follows.

- 99 037533

Obtaining 1 - (5-bromo-3-fluoropyridin-2-yl) -N - ((1S, 3R) -1 - (5-bromo-3-fluoropyridin-2-yl) -2 - ((1 fluorocyclopropyl) methyl ) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl) methanimine

Bg | ^G ό .¾¾

5-Bromo-3-fluoropicolinaldehyde (1.09 g, 5.33 mmol) was added to (R) -3- (2 - (((1fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (630 mg, 2, 67 mmol) and water (0.240 ml, 13.33 mmol) in acetic acid (12 ml). The resulting solution was stirred at 70 ° C for 2 h. The reaction mixture was concentrated and diluted with EtOAc (50 ml) and washed sequentially with saturated aqueous NaHCO ₃ solution (50 ml). The organic layer was dried with MgSO ₄ , filtered and evaporated to give 1- (5-bromo-3-fluoropyridin-2-yl) -N - ((1S, 3R) -1 (5-bromo-3-fluoropyridine-2- yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl) methanimine as crude product (1.53 g, 2.51 mmol) ...

Mass / charge: ES + [M + H] ⁺ = 607.

Obtaining (1S, 3R) -1- (5-bromo-3-fluoropyridin-2-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinoline-6- amine

Bg

I F

Hydroxylamine hydrochloride (0.174 g, 2.51 mmol) was added to 1- (5-bromo-3-fluoropyridin-2-yl) -N ((1S, 3R) -1- (5-bromo-3-fluoropyridine-2- yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinolin-6-yl) methanimine (1.53 g, 2.51 mmol) and potassium acetate (0, 62 g, 6.27 mmol) in MeOH (12 ml). The resulting solution was stirred at 20 ° C for 5 h. The reaction mixture was concentrated and diluted with DCM (75 ml) and washed sequentially with NaOH (2 M, 75 ml) and saturated aqueous sodium chloride solution (50 ml). The organic layer was dried with MgSO ₄ , filtered and evaporated to give the crude product, which was purified by flash chromatography on silica, elution gradient 0 to 20% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give (1S, 3R) -1- (5-6pom-3fluoropyridin-2-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1, 2,3,4-tetrahydroisoquinoline-6-amine (0.672 g, 63%) as a colorless gum.

1H NMR (500 MHz, CDCl3, 27 ° C) 0.45-0.61 (2H, m), 0.92-1.06 (2H, m), 1.07 (3H, d), 2.07 (3H, s), 2.51 (1H, dd), 2.59 (1H, dd), 2.85 (1H, dd), 3.13 (1H, dd), 3.52 (2H, s) , 3.64-3.75 (1H, m), 5.35 (1H, s), 6.48 (2H, s), 7.52 (1H, dd), 8.36 (1H, dd);

mass / charge: ES ⁺ [M + H] ⁺ = 422/424.

Obtaining (6S, 8R) -6- (5-bromo-3-fluoropyridin-2-yl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [ 4,3-1] isoquinoline

(1S, 3R) -1- (5-Bromo-3-fluoropyridin-2-yl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine (0.672 g, 1.59 mmol) in propionic acid (6.63 ml) was cooled to -17 ° C (dry ice / acetone). Sodium nitrite (0.110 g, 1.59 mmol) in water (1.33 ml) was added dropwise and the reaction mixture was stirred at -17 ° C for 30 min. The reaction mixture was diluted with ice-cold toluene (30 ml), stirred at 0 ° C for 15 minutes, then at room temperature for 45 minutes. The reaction mixture was washed with water (2x25 ml), the combined aqueous phases were washed with EtOAc (25 ml), the combined organics were washed with saturated aqueous sodium chloride solution (50 ml), dried (MgSO ₄ ), filtered and the filtrate was evaporated to an orange-brown oil. The crude material was purified by flash chromatography on silica, gradient elution 0-50% EtOAc in heptane to give (6S, 8R) -6- (5-6pom-3fluoropyridin-2-yl) -7 - ((1-fluorocyclopropyl ) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-Aisoquinoline (0.310 g, 45%) as an orange solid.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.50-0.59 (2H, m), 1.03 (2H, dd), 1.13 (3H, d), 2.72 (1H , dd), 2.91 (1H,

- 100 037533 dd), 3.16 (1H, dd), 3.27 (1H, dd), 3.77-3.90 (1H, m), 5.50 (1H, s), 6.80 ( 1H, d), 7.20 (1H, dd), 7.56 (1H, dd),

8.07 (1H, d), 8.36 (1H, dd);

mass / charge: ES + [M + H] ⁺ = 433.

Example 19.

Preparation of N- (4 - ((6S, 8R) -7- (2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl ) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine

DMF (2 ml) and DIPEA (0.074 ml, 0.42 mmol) were added sequentially to the flask, which was loaded with N- (4 - ((6S, 8R) -7- (2,2-difluoropropyl) -8-methyl -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) -3-methoxyphenyl) azetidine-3-amine (75 mg, 0.17 mmol). Then 1-fluoro-3-iodopropane (31.9 mg, 0.17 mmol) in DMF (0.1 ml) was added and stirring was continued for 2 h. The reaction was stopped, the reaction mixture was diluted with saturated aqueous sodium chloride solution and the compound was extracted in EtOAc (x3). The combined extracts were washed with water and dried over sodium sulfate, filtered and concentrated under reduced pressure to give a film. This material was purified by flash chromatography on silica eluting 2% to 10% (methanol containing 1% ammonium hydroxide) in DCM to give N- (4 - ((6S, 8R) -7- (2,2-difluoropropyl ) -8-methyl6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidine-3amine (43 mg, 51%).

1H NMR (400 MHz, DMSO-d6, 27 ° С) 1.00 (3H, d), 1.45-1.70 (5H, m), 2.44 (2H, t), 2.55-2 , 66 (1H, m), 2.66-2.72 (2H, m), 2.79 (1H, dd), 2.87-3.01 (1H, m), 3.10 (1H, br dd), 3.38-3.53 (1H, m), 3.56-3.67 (2H, m), 3.77 (3H, s), 3.85-3.97 (1H, m) , 4.43 (2H, dt), 5.21 (1H, s), 5.88 (1H, dd), 6.00 (1H, d), 6.16 (1H, d), 6.35 ( 1H, d), 6.64 (1H, d), 7.17 (1H, d), 8.02 (1H, s), 12.92 (1H, d);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 502.

N- (4 - ((6S, 8R) -7- (2,2-Difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6yl) -3-methoxyphenyl) azetidin-3-amine starting material was prepared according to the procedures below.

Preparation of 2,2-difluoropropyl trifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (3.29 ml, 19.5 mmol) was added dropwise to a solution of 2,2-difluoropropan-1-ol (1.7 g, 18 mmol) in DCM (40 ml) at -10 ° C (bath with salt / ice). Then 2,6-dimethylpyridine (2.5 ml, 21 mmol) was added and the reaction mixture was stirred for 1 hour under these conditions. The reaction mixture was then washed with water (x2) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure (vacuum ~ 200 mbar) to give 2,2-difluoropropyl trifluoromethanesulfonate (2.1 g, 52%) as a red oil ...

1H NMR (400 MHz, CHLOROFORM-d, 27 ° C) 4.48 (2H, t), 1.73 (3H, t).

Preparation of (R) -3- (2 - ((2,2-difluoropropyl) amino) propyl) -2-methylaniline

2,2-Difluoropropyl trifluoromethanesulfonate (1.68 g, 7.37 mmol) was added to a stirred solution of (R) -3- (2-aminopropyl) -2-methylaniline (1.1 g, 6.7 mmol) and DIPEA (1 , 52 ml, 8.71 mmol) in 1,4-dioxane (20 ml). The reaction mixture was heated at 65 ° C for 3 hours before being cooled to room temperature and then concentrated under reduced pressure. The resulting residue was dissolved in EtOAc (30 ml) and washed with saturated aqueous NaHCO ₃ solution. The aqueous layer was extracted with EtOAc (20 ml) and the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting red oil was purified by flash chromatography on silica, elution gradient from 30 to 90% ethyl acetate in hexanes, to give (R) -3- (2 - ((2,2-difluoropropyl) amino) propyl) -2-methylaniline (1.02 g, 63%) as gum.

1H NMR (500 MHz, DMSO-d ₆ , 27 ° C) 0.90 (3H, d), 1.55 (3H, t), 1.75 (1H, br s), 1.97 (3H, s ), 2.29-2.38 (1H, m), 2.69-2.76 (2H, m), 2.86 (2H, br t), 4.69 (2H, s), 6.34 (1H, d), 6.47 (1H, d), 6.77 (1H, t);

mass / charge: (eS ⁺ ) [M + H] ⁺ = 439.

- 101 037533

Preparation of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine

Br

4-Bromo-2-methoxybenzaldehyde (1.07 g, 4.95 mmol) was added to a solution of (R) -3- (2 - ((2,2 difluoropropyl) amino) propyl) -2-methylaniline (0.600 g, 2, 48 mmol) in AcOH (12 ml) and water (0.223 g, 12.4 mmol) and the reaction mixture was heated at 80 ° C for 18 h. After cooling, the volatiles were concentrated under reduced pressure and the resulting residue was dissolved in EtOAc. The solution was neutralized by washing with saturated aqueous NaHCO ₃ solution. The organic layer was combined with aqueous HCl (1N) and the biphasic mixture was stirred at room temperature for 30 minutes. Then the layers were separated and the organic layer was washed with aqueous HCl (1N). The combined aqueous layers were extracted with EtOAc and then made basic by adding solid K ₂ CO ₃ . Then the organic layer was extracted with EtOAc (x2) and the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 10% to 60% ethyl acetate in hexanes, to give (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2 (2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (0.576 g, 53%) as gum.

1H NMR (400 MHz, DMSO-d ₆ , 27 ° C) 0.95 (3H, d), 1.54 (3H, t), 1.93 (3H, s), 2.27-2.43 ( 2H, m), 2.57-2.74 (1H, m), 2.76-2.98 (1H, m), 3.19-3.26 (1H, m), 3.84 (3H, s), 4.63 (2H, s), 5.12 (1H, s), 6.26 (1H, d), 6.38 (1H, d), 6.58 (1H, d), 6, 94 (1H, dd), 7.16 (1H, d);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 439.

N - ((1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2,2-difluoropropyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinolin-6-yl ) acetamide (0.081 g, 7%) as a gum.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro3H-pyrazolo [4,3-f ] isoquinoline

Br

Sodium nitrite (0.049 g, 0.72 mmol) as a solution in water (0.750 ml) was added dropwise to a cooled solution of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -2- (2.2 -difluoropropyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine (0.30 g, 0.68 mmol) in propionic acid (2.5 ml) at -15 ° C and the reaction mixture was stirred within 1 hour under such conditions. Ice-cold EtOAc (10 ml) was added followed by portionwise addition of saturated aqueous NaHCO ₃ (10 ml). The layers were separated and the organic layer was washed with saturated aqueous NaHCO ₃ solution. The combined aqueous layers were extracted with EtOAc and all organic layers were combined, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 20 to 70% ethyl acetate in hexanes, to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoropropyl) - 8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (0.23 g, 75%) as gum.

1H NMR (500 MHz, DMSO-d ₆ , 27 ° С) 1.01 (3H, d), 1.53 (3H, t), 2.84 (1H, dd), 2.96-3.09 ( 1H, m), 3.14 (1H, br dd), 3.36-3.48 (1H, m), 3.89 (3H, s), 5.32 (1H, s), 6.64 ( 2H, apparent t), 6.94 (1H, dd), 7.16-7.27 (2H, m), 8.06 (1H, s), 12.99 (1H, br s). 1H blocked by DMSO.

Mass / charge: (ES ⁺ ) [M + H] ⁺ = 450.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoropropyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 (2,2-difluoropropyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4,3f] isoquinoline

- 102 037533

DCM (7 ml) was added to the flask, which was charged with (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7 (2,2-difluoropropyl) -8-methyl-6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.3 g, 0.67 mmol) and 4-methylbenzenesulfonic acid hydrate (0.025 g, 0.13 mmol). 3,4-dihydro-2H-pyran (0.084 g,

1.0 mmol) and the reaction mixture was stirred at room temperature for 18 hours.

the mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure. The resulting residue was purified by flash chromatography eluting with 5-40% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoropropyl) -8- methyl 3- (tetrahydro-2H-pyran2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (175 mg, 49%) as gum.

¹ H NMR (400 MHz, CHLOROFORM-d, 27 ° С) 0.99-1.13 (3H, br s), 1.48-1.88 (6H, m), 2.01-2.08 ( 1H, m), 2.08-2.18 (1H, m), 2.48-2.70 (2H, m), 2.82 (1H, dt), 2.88-3.04 (1H, m), 3.20 (1H, br d), 3.55-3.67 (1H, m), 3.633.73 (1H, m), 3.88 (3H, s), 3.94-3, 98 (1H, m), 5.42 (1H, br s), 5.63 (1H, dt), 6.64 (1H, dd), 6.73 (1H, dd), 6.86 (1H, dd), 7.03 (1H, t), 7.25-7.29 (1H, m), 7.98 (1H, d);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 534.

Also (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoropropyl) -8-methyl-2- (tetrahydro2H-pyran-2-yl) -6,7, 8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline (68 mg, 19%) as gum.

¹ H NMR (400 MHz, CHLOROFORM-d, 27 ° С) 1.02-1.07 (3H, m), 1.42-1.53 (3H, m), 1.59-1.81 (3H, m), 2.03-2.09 (1H, m), 2.18-2.25 (2H, m), 2.59 (1H, qd), 2.69 (1H, dd), 2.86 -2.98 (1H, m), 3.02-3.14 (1H, m), 3.463.57 (1H, m), 3.72-3.82 (1H, m), 3.87-3 , 90 (3H, m), 4.11-4.16 (1H, m), 5.31 (1H, s), 5.61-5.68 (1H, m), 6.60 (1H, dd ), 6.72 (1H, dd), 6.88 (1H, dt), 7.01-7.05 (1H, m), 7.36 (1H, d), 8.09 (1H, s) ;

mass / charge: (ES ⁺ ) [M + H] ⁺ = 534.

Preparation of tert-butyl-3 - ((4 - ((6S, 8R) -7- (2,2-difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine-1-carboxylate

Dioxane (3.5 ml) was added to a flask, which was charged with CS ₂ CO ₃ (213 mg, 0.65 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (85 mg, 0.49 mmol) and (6S , 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (175 mg, 0.33 mmol). The reaction flask was evacuated and refilled with nitrogen (x3). BrettPhos 3rd generation precatalyst (30 mg, 0.03 mmol) was added and the flask was evacuated again and refilled with nitrogen (x3). The reaction mixture was heated at 100 ° C for 4 hours. The reaction mixture was cooled to room temperature, filtered and concentrated under reduced pressure. The resulting crude resin was purified by flash chromatography on silica eluting with 15-70% ethyl acetate in hexanes to give tert-butyl 3 - ((4 - ((6S, 8R) -7 (2,2-difluoropropyl) - 8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine -1-carboxylate (152 mg, 74%) as a film.

1H NMR (400 MHz, CHLOROFORM-d, 27 ° С) 1.05 (3H, d), 1.42 (9H, s), 1.44-1.57 (3H, m), 1.57-1 , 63 (1H, m), 1.66-1.77 (2H, m), 2.04-2.08 (1H, m), 2.09-2.18 (1H, m), 2.47 -2.69 (2H, m), 2.79 (1H, dt), 2.84-2.99 (1H, m), 3.16 (1H, br dd), 3.48-3.61 ( 1H, m), 3.69 (3H, dt), 3.83 (3H, s), 3.94-4.04 (2H, m), 4.15 (1H, br s), 4.20- 4.29 (2H, m), 5.32 (1H, s), 5.63 (1H, dt), 5.86 (1H, dd), 6.07 (1H, t), 6.55 (1H , dd), 6.78 (1H, dd), 7.20-7.23 (1H, m), 7.98 (1H, d);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 626.

Preparation of N- (4 - ((6S, 8R) -7- (2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl ) -3-methoxyphenyl) azetidine-3-amine

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Methanol (1 ml) and then HCl in dioxane (4 M; 1 ml, 4 mmol) were sequentially added to the flask, into which tert-6utyl-3 - ((4 - ((6S, 8R) -7- (2 , 2-difluoropropyl) -8-methyl-3- (tetrahydro-2H-pyran2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3 α-methoxyphenyl) amino) azetidine-1carboxylate (140 mg, 0.22 mmol). After 2 hours, the reaction mixture was concentrated under reduced pressure and the resulting residue was purified using an SCX-2 cartridge that had been pretreated with methanol. The compound was eluted first with methanol and then with ammonia in methanol (3N). Product fractions were concentrated under reduced pressure to give N- (4 - ((6S, SR) -7- (2,2 difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4.3 -f] isoquinolin-6-yl) -3-methoxyphenyl) azetidin-3-amine (91 mg, 92%) as gum. The product was used in the next step without further purification.

¹ H NMR (400 MHz, DMSO-d6, 27 ° C) 1.00 (3H, d), 1.52 (3H, t), 2.55-2.59 (1H, m), 2.78 ( 1H, dd), 2.852.99 (1H, m), 3.09 (1H, dd), 3.39-3.50 (1H, m), 3.55-3.66 (2H, m), 3 , 77 (3H, s), 4.03-4.14 (1H, m), 5.21 (1H, s),

5.85 (1H, dd), 6.05 (1H, br d), 6.14 (1H, d), 6.34 (1H, d), 6.63 (1H, d), 7.17 ( 1H, d), 8.02 (1H, s), 12.93 (1H, br s). 3H was not observed, mass / charge: (ES ⁺ ) [M + H] ⁺ = 442.

Example 20.

Preparation of N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidine-3-amine

DMF (2 ml) and DIPEA (0.050 ml, 0.28 mmol) were sequentially added to the flask, which was loaded with N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl ) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (0.051 g, 0.11 mmol). 1-Fluoro-3-iodopropane (0.021 g, 0.11 mmol) was added as a solution in DMF (0.2 ml) and after 2 hours the reaction mixture was diluted with saturated aqueous sodium hydrogen carbonate solution. The mixture was extracted with ethyl acetate (x3) and the combined organic layers were washed with water, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 2 to 10% (methanol containing 1% ammonium hydroxide) in DCM. Product fractions were combined, concentrated under reduced pressure and the resulting residue was purified by flash silica chromatography using the above conditions to give N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine (0.023 d, 40%).

¹ H NMR (CHLOROFORM-d, 27 ° C) 0.50 (2H, br d), 0.88-1.05 (2H, m), 1.12 (3H, br d), 1.45 (3H , t), 1.751.87 (2H, m), 2.59-2.73 (3H, m), 2.87-3.04 (3H, m), 3.04-3.15 (1H, m ), 3.41 (1H, br dd), 3.73-3.90 (3H, m), 3.93-4.18 (4H, m), 4.52 (2H, dt), 5.37 (1H, br s), 5.99 (1H, dd), 6.13 (1H, br d), 6.80-6.89 (2H, m), 7.14 (1H, d), 8, 06-8.08 (1H, m); 10.02 (1H, br s);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 510.

The procedures used to obtain N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4, 3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine are described below.

Preparation of (1S, 3R) -1- (4-bromo-2-ethoxyphenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4tetrahydroisoquinoline-6-amine

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4-Bromo-2-ethoxybenzaldehyde (0.775 g, 3.39 mmol) was added to a solution of (R) -3- (2 - (((1fluorocyclopropyl) methyl) amino) propyl) -2-methylaniline (0.400 g, 1.69 mmol) in AcOH (9 ml) and water (0.152 g, 8.46 mmol). The reaction mixture was heated at 80 ° C for 18 h. After cooling, the reaction mixture was concentrated under reduced pressure and then the residue was dissolved in EtOAc and washed with saturated aqueous NaHCO ₃ solution. The layers were separated and the organic phase was combined with aqueous hydrochloric acid (1N). After stirring the biphasic mixture for 30 min, the layers were separated. The organic layer was washed with aqueous HCl (1N) and the combined aqueous layers were extracted with EtOAc (x2). The combined aqueous layers were then podslushivaet by adding solid K ₂ CO ₃ and was extracted with EtOAc (x2). The organic extracts were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 10% to 60% ethyl acetate in hexanes, to give (1S, 3R) -1- (4-6pom-2ethoxyphenyl) -2 - ((1-fluorocyclopropyl) methyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (0.48 g, 63%) as a solid.

¹ H NMR (400 MHz, DMSO-d6, 27 ° С) 0.40-0.59 (2H, m), 0.83-0.93 (2H, m), 0.94 (3H, d), 1 , 35 (3H, t), 1.93-1.96 (3H, m), 2.45-2.53 (2H, m), 2.80-2.93 (2H, m), 3.57 (1H, br d), 4.14 (2H, q), 4.58 (2H, s), 5.15 (1H, s), 6.26 (1H, d), 6.35 (1H, d ), 6.86 (1H, d), 6.95 (1H, dd), 7.15 (1H, d);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 447.

Obtaining (6S, 8R) -6- (4-bromo-2-ethoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9 tetrahydro-3H-pyrazolo [4,3 -f] isoquinoline

Sodium nitrite (0.039 g, 0.56 mmol) as a solution in water (0.750 ml) was added dropwise to a cooled solution of (1S, 3R) -1- (4-bromo-2-ethoxyphenyl) -2 - ((1- fluorocyclopropyl) methyl) -3,5-dimethyl1,2,3,4-tetrahydroisoquinolin-6-amine (0.240 g, 0.54 mmol) in propionic acid (3.0 ml) at -15 ° C and the reaction mixture was maintained at under these conditions for 1 h. Then ice-cold EtOAc (10 ml) was added followed by portionwise addition of saturated aqueous NaHCO ₃ solution (15 ml). Once the addition was complete and gas evolution ceased, the layers were separated. The organic layer was washed with saturated aqueous NaHCO ₃ (x2) and the combined aqueous layers were extracted with EtOAc (x2). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography, elution gradient from 20 to 60% ethyl acetate in hexanes, to give (6S, 8R) -6- (4bromo-2-ethoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl- 6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (0.145 g, 59%) as gum.

¹ H NMR (500 MHz, DMSO-d6, 27 ° С) 0.45 (1H, br s), 0.57 (1H, br s), 0.83-0.98 (2H, m), 1.00 (3H, d), 1.37 (3H, t), 2.54-2.63 (1H, m), 2.87-3.00 (2H, m), 3.27-3.32 (1H , m), 3.74 (1H, br d), 4.13-4.23 (2H, m), 5.32 (1H, s), 6.67 (1H, d), 6.94 (1H , s), 6.96-6.99 (1H, m), 7.19 (1H, d), 7.22 (1H, d), 8.06 (1H, s), 12.96 (1H, s);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 458.

Obtaining (6S, 8R) -6- (4-bromo-2-ethoxyphenyl) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

Br

DCM (3 ml) and 3,4-dihydro-2H-pyran (35.8 mg, 0.43 mmol) were added to a flask containing (6S, 8R) -6- (4-bromo-2-ethoxyphenyl ) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro3H-pyrazolo [4,3-f] isoquinoline (130 mg, 0.28 mmol) and 4-methylbenzenesulfonic acid hydrate acid

- 105 037533 (10.79 mg, 0.06 mmol). The reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting brown gum was purified by flash chromatography on silica, elution gradient from 5% to 40% ethyl acetate in hexanes, to give (6S, 8R) -6- (4-bromo-2-ethoxyphenyl) 7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (147 mg, 96%) as dry films.

¹ H NMR (500 MHz, CHLOROFORM-d, 27 ° C) 0.42-0.51 (2H, m), 0.94-1.03 (2H, m), 1.07-1.12 (3H, m), 1.49 (3H, t), 1.61-1.68 (1H, m), 1.73-1.79 (2H, m), 2.02-2.10 (1H, m) , 2.12-2.20 (1H, m), 2.53-2.63 (2H, m), 2.93 (1H, dt), 3.10 (1H, ddd), 3.38-3 , 46 (1H, m), 3.65-3.77 (1H, m), 3.83-3.92 (1H, m), 4.00-4.07 (1H, m), 4.124.19 (2H, m), 5.43 (1H, s), 5.66 (1H, ddd), 6.81 (1H, d), 6.89-6.93 (1H, m), 6.95- 7.00 (1H, m), 7.03-7.06 (1H, m), 7.25 (1H, dd), 8.03 (1H, s);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 542.

Preparation of tert-butyl-3 - ((3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) amino) azetidine-1-carboxylate

Dioxane (2.7 ml) was added to a flask containing (6S, 8R) -6- (4-bromo-2-ethoxyphenyl) 7 - ((1-fluorocyclopropyl) methyl) -8-methyl-3- ( tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (140 mg, 0.26 mmol), Cs ₂ CO ₃ (168 mg, 0, 52 mmol) and tert-butyl 3aminoazetidine-1-carboxylate (67 mg, 0.39 mmol). The reaction flask was evacuated and filled with N ₂ (x3). BrettPhos 3rd generation precatalyst (23 mg, 0.030 mmol) was added and the flask was evacuated and filled with nitrogen (x3). The reaction mixture was heated at 90 ° C for 1 h and then at 90 ° C for 44 h. The reaction mixture was cooled to room temperature, filtered and concentrated under reduced pressure. The resulting resin was purified by flash chromatography, elution gradient from 30 to 100% ethyl acetate in hexanes, to give tert-butyl 3 - ((3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8 -methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) amino) azetidine-1-carboxylate (86 mg, 53%) as gum.

¹ H NMR (500 MHz, CHLOROFORM-d, 27 ° С) 0.39-0.49 (2H, m), 0.87-0.95 (2H, m), 1.06 (3H, d), 1 , 38-1.45 (12H, m), 1.57-1.62 (1H, m), 1.68-1.74 (2H, m), 1.98-2.06 (1H, m) , 2.07-2.15 (1H, m), 2.49-2.63 (2H, m), 2.85 (1H, dt), 3.05 (1H, br t), 3.29- 3.35 (1H, m), 3.63-3.73 (3H, m), 3.75-3.83 (1H, m), 3.92 (1H, br d), 3.99 (1H , br d), 4.05 (2H, q), 4.14 (1H, br dd), 4.18-4.26 (2H, m), 5.33 (1H, s), 5.58- 5.63 (1H, m), 5.90 (1H, br d), 6.02-6.06 (1H, m), 6.76 (1H, dd), 6.80 (1H, d), 7.19 (1H, d), 7.97 (1H, s);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 634.

Preparation of N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine

Methanol (0.5 ml) was added to the flask, which was charged with tert-butyl-3 - ((3-ethoxy-4 - ((6S, 8R) -7 ((1-fluorocyclopropyl) methyl) -8-methyl- 3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) phenyl) amino) azetidine-1-carboxylate (0.080 g , 0.13 mmol). Hydrochloric acid in dioxane (4M; 0.5 ml, 2 mmol) was added and stirring was continued for 2 hours. The reaction mixture was then concentrated under reduced pressure and the resulting residue was purified using an SCX-2 cartridge pretreated with methanol, eluting first with methanol and then with a solution of ammonia in methanol (3N) to obtain N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1fluorocyclopropyl) methyl) -8-methyl-6,7,8,9 -tetrahydro-3H-pyrαzolo [4,3-f] isoquinolin-6-yl) phenyl) azetidine-3-amine (0.057 g, 99%) as a solid.

- 106 037533

1H NMR (500 MHz, CHLOROFORM-d, 27 ° С) 0.44-0.55 (2H, m), 0.90-1.00 (2H, m), 1.12 (3H, d), 1, 45 (3H,

t), 2.61-2.71 (1H, m), 2.92 (1H, br dd), 3.10 (1H, br dd), 3.40 (1H, br dd), 3.51- 3.58 (2H, m), 3.83-3.90 (1H, m),

3.91-4.02 (2H, m), 4.03- 4.16 (3H, m), 4.36 (1H, sxt), 5.38 (1H, s), 5.98 (1H, dd), 6.12 (1H, br d), 6.77-6.89 (2H,

m), 7.13 (1H, d), 8.07 (1H, s), 10.09 (1H, br s). One H was not observed, and it appears to be overlapped by a peak of water, mass / charge: (ES +) [M + H] ⁺ = 450.

Example 21.

Preparation of N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl ) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine

Hydrochloric acid in dioxane (4 M; 1.23 ml, 5.2 mmol) was added dropwise to a stirred solution of tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) - 8-methyl-2- (tetrahydro-2H-pyran-2-yl) 6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenylamino) azetidine- 1-carboxylate (316 mg, 0.52 mmol) in MeOH (5 ml). The mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure and the resulting residue was dissolved in MeOH. An excess of macroporous tetraalkylammonium carbonate resin (Aldrich; 18-50 mesh; loading 2.5-3.5 mmol / g N) was added and the mixture was stirred at room temperature for 5 minutes. The mixture was filtered and the filtrate was dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8, 9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) azetidin-3-amine gum (220 mg). This material was directly used in the next step without further purification, mass / charge: (ES +) [M + H] ⁺ = 428. A mixture of N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) - 8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) azetidin-3-amine (220 mg, 0.51 mmol), 1- fluoro-3-iodopropane (106 mg, 0.57 mmol) and DIPEA (0.270 ml, 1.54 mmol) in DMF (5 ml) were stirred at room temperature for 18 h. The reaction mixture was diluted with DCM and washed with saturated aqueous solution of ammonium chloride. The aqueous layer was extracted with DCM and the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (Xbridge C18 column, 19x150 mm; 5 μm; flow rate: 20 ml / min.), Eluting with 40-80% acetonitrile in water containing 0.2% ammonium hydroxide. The product fractions were concentrated under reduced pressure and the resulting residue was purified by preparative SFC (2-ethylpyridine column, 19x150 mm, 5 μm; flow rate: 75 ml / min; column temperature: 40 ° C; outlet pressure: 100 bar) eluting with 15% methanol containing 0.2% ammonium hydroxide in carbon dioxide to give N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7 , 8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine (82 mg, 33% in two steps) in the form of a white solid.

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.03 (3H, d), 1.55-1.74 (2H, m), 2.41-2.47 (2H, m), 2.53-2.83 (4H, m), 2.86-3.04 (1H, m), 3.10 (1H, br dd), 3.32-3.43 (1H, m), 3 , 58-3.66 (2H, m), 3.82 (3H, s), 3.86-3.98 (1H, m), 4.45 (2H, dt), 5.21 (1H, s ), 5.89 (1H, t), 5.90 (1H, dd), 6.01 (1H, d), 6.19 (1H, d), 6.35 (1H, d), 6.67 (1H, d), 7.19 (1H, d), 8.03 (1H, s), 12.92-12.96 (1H, m);

mass / charge: ES ⁺ [M + H] ⁺ = 488.

tert-Butyl-3 - ((4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) amino) azetidine-1-carboxylate starting material was prepared according to the following procedures.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2Hpyran-2-yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline and (6S, 8R) -6-th-bromo-2-methoxyphenyl) -7 (2,2-difluoroethyl) -8-methyl-2- ( tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4,3f] isoquinoline

- 107 037533

(6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3-f] isoquinoline (986 mg, 2.26 mmol), 4-methylbenzenesulfonic acid hydrate (43 mg, 0.23 mmol), 3,4-dihydro-2H-pyran (0.31 ml, 3.4 mmol) and DCM. The reaction mixture was heated at 80 ° C under microwave conditions (300 W) for 20 min. The reaction mixture was then cooled and diluted with DCM before washing with saturated aqueous sodium hydrogen carbonate solution. The layers were separated and the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 70% EtOAc in hexanes, to give the more rapidly eluting (6S, 8R) -6- (4bromo-2-methoxyphenyl) -7- (2,2- difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (457 mg, 39%) as orange solid matter.

1H NMR (300 MHz, CHLOROFORM-d, 27 ° C) 1.04-1.22 (3H, m), 1.59-1.86 (3H, m), 2.04-2.23 (2H, m), 2.53-3.31 (5H, m), 3.40-3.57 (1H, m), 3.68-3.79 (1H, m), 3.97 (3H, s) , 4.00-4.09 (1H, m), 5.41 (1H, br s), 5.69 (1H, br s), 5.66-5.73 (1H, m), 6.67 (1H, dd), 6.80 (1H, dd), 6.92 (1H, dd), 7.06-7.12 (1H, m), 7.29-7.37 (1H, m), 8.03 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 612.

The more slowly eluted (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2difluoroethyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) -6 , 7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline (627 mg, 57%) as a beige powder.

1H NMR (300 MHz, CHLOROFORM-d, 27 ° C) 1.12 (3H, br d), 1.64-1.87 (3H, m), 2.03-2.17 (1H, m), 2.182.29 (2H, m), 2.61-2.83 (2H, m), 2.89-3.15 (2H, m), 3.43 (1H, br s), 3.75-3 , 86 (1H, m), 3.95 (3H, s), 4.13-4.20 (1H, m), 5.32 (1H, s), 5.79 (1H, br t), 5 , 66-5.73 (1H, m), 6.63-6.77 (2H, m), 6.93 (1H, d), 7.09 (1H, d), 7.43 (1H, d ), 8.14 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 612.

Preparation of tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-2- (tetrahydro-2H-pyran-2-yl) 6,7,8, 9-tetrahydro-2H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenylamino) azetidine-1-carboxylate

(6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2 difluoroethyl) -8-methyl-2- (tetrahydro-2H-nyran-2-yl) - 6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinoline (310 mg, 0.60 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (154 mg, 0.89 mmol ), CS ₂ CO ₃ (388 mg, 1.19 mmol) and BrettPhos 3rd generation precatalyst (54.0 mg, 0.06 mmol). The vial was degassed and filled with nitrogen (x2). The vial was degassed again and refilled with 1,4-dioxane (6 ml) and nitrogen. The vial was degassed again and refilled with nitrogen. The mixture was heated under microwave conditions (300 W, 110 ° C) for 3.5 hours. The reaction mixture was diluted with DCM and washed with saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 20 to 60% EtOAc in hexanes, to give tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) - 8methyl-2- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-2H-pyrazolo [4,3-f] isoquinolin-6-yl) -3methoxyphenylamino) azetidine-1-carboxylate (322 mg, 88%) as an off-white solid.

1H NMR (300 MHz, CHLOROFORM-d, 27 ° C) 1.08 (3H, br d), 1.39-1.43 (9H, m), 1.58-1.81 (3H, m), 2.06 (1H, br dd), 2.13-2.25 (2H, m), 2.58-2.71 (1H, m), 2.82-3.11 (2H, m), 3 , 34-3.50 (1H, m), 3.66-3.81 (3H, m), 3.86 (3H, s), 3.90-4.01 (1H, m), 4.08 -4.20 (3H, m), 4.19-4.29 (2H, m), 5.22 (1H, s), 5.76 (1H, t), 5.64 (1H, t), 5.88 (1H, br d), 6.09 (1H, d), 6.59 (1H, br t), 6.67 (1H, d), 7.37 (1H, br d), 8, 08 (1H, s);

- 108 037533 mass / charge: ES ⁺ [M + H] ⁺ = 612.

Example 22.

Obtaining (6S, 8R) -7- (2,2-difluoroethyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

HCl in dioxane (4 M; 1.02 ml, 4.08 mmol) was added dropwise to a stirred solution of tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl -3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate ( 250 mg, 0.41 mmol) in MeOH (4 ml). The mixture was stirred at room temperature for 18 hours. The reaction mixture was then concentrated under reduced pressure and the resulting residue was dissolved with MeOH. An excess of macroporous tetraalkylammonium carbonate resin (Aldrich; 18-50 mesh; loading 2.5-3.5 mmol / g N) was added and the mixture was stirred at room temperature for 5 min before being dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude (6S, 8R) -6- (4- (azetidin-3-yloxy) -2-methoxyphenyl) -7- (2,2-difluoroethyl) -8-methyl 6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (179 mg), which was used directly in the next step without further purification, mass / charge: ES + [M + H] ⁺ = 429. Mixture (6S, 8R) -6 - (4 (azetidin-3-yloxy) -2-methoxyphenyl) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline ( 178 mg, 0.42 mmol), 1-fluoro-3-iodopropane (94 mg, 0.50 mmol) and DIPEA (0.218 ml, 1.25 mmol) in DMF (4 ml) were stirred at room temperature for 18 h Then the reaction mixture was diluted with DCM and washed with saturated aqueous ammonium chloride solution. The layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by preparative SFC (2-ethylpyridine column; 19 mm x 150 mm; 5 μm; flow rate: 75 ml / min; column temperature: 40 ° C; outlet pressure: 100 bar) eluting with 15 % (methanol containing 0.2% ammonium hydroxide) in carbon dioxide to give (6S, 8R) -7- (2,2-difluoroethyl) -6- (4 - ((1- (3-fluoropropyl) azetidine-3 -yl) oxy) -2-methoxyphenyl) 8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (70 mg, 35% over two steps) as a white solid ...

1H NMR (300 MHz, DMSO-de, 27 ° C) 1.02 (3H, d), 1.61-1.84 (2H, m), 2.55-2.84 (3H, m), 2 , 90-3.13 (2H, m), 3.18-3.43 (4H, m), 3.87 (3H, s), 3.89-4.00 (2H, m), 4.45 (2H, dt), 4.81 (1H, quin), 5.28 (1H, s), 5.74-6.16 (1H, m), 6.19 (1H, dd), 6.49- 6.54 (2H, m), 6.66 (1H, d), 7.21 (1H, d), 8.04 (1H, s), 12.98 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 489.

Procedures used to prepare tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6, 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate starting material are described below.

Preparation of tert-butyl-3- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate

(6S, 8R) -6- (4-bromo-2-methoxyphenyl) -7- (2,2 difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (259 mg, 0.50 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (259 mg, 1.49 mmol ), cesium carbonate (324 mg, 1.00 mmol) and RockPhos 3rd generation pre-catalyst (42 mg, 0.050 mmol). The vial was evacuated and filled with nitrogen (x2). The vial was then evacuated and filled with toluene (3 ml). The vial was evacuated and filled with nitrogen. The mixture was heated at 110 ° C. for 4 hours. The reaction mixture was diluted with DCM and washed with saturated aqueous solution of sodium hydroxide 109 037533. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting from 0 to 60% EtOAc in hexanes to give tert-butyl-3- (4 ((6S, 8R) -7- (2,2-difluoroethyl) -8- methyl 3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -3-methoxyphenoxy) azetidine-1-carboxylate (220 mg, 72%) as a white foam solid.

1H NMR (400MHz, DMSO-d6, 100 ° C) 1.03-1.09 (3H, m), 1.40 (9H, s), 1.56-1.64 (2H, m), 1, 67-1.84 (1H, m), 1.93-2.01 (1H, m), 2.02-2.12 (1H, m), 2.34-2.46 (1H, m), 2.59-2.74 (1H, m), 2.84 (1H, dt), 2.93-3.06 (1H, m), 3.13-3.23 (1H, m), 3, 40-3.50 (1H, m), 3.65-3.74 (1H, m), 3.76-3.84 (2H, m), 3.84-3.91 (4H, m), 4.21-4.28 (2H, m), 4.97 (1H, tt), 5.33 (1H, s), 5.67-6.00 (1H, m), 5.74 (1H, dd), 6.23 (1H, ddd), 6.54 (1H, d), 6.66 (1H, t), 6.73 (1H, d), 7.35 (1H, d), 8, 04 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 613.

Examples 23 and 24.

Preparation of individual diastereoisomers 3 - ((6S, 8R) -6- (2,6-difluoro-4- (1- (3-fluoropropyl) azetidine-3ylamino) phenyl) -8-methyl-8,9-dihydro-3H- pyrazolo [4,3-f] isoquinoline-7 (6H) -yl) -2-fluoro-2-methylpropane-1ol

Tetrabutylammonium fluoride in THF (1 M; 0.114 ml, 0.11 mmol) was added dropwise using a syringe to a stirred solution of N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3fluoropropyl ) azetidine-3-amine (82 mg, 0.08 mmol) in THF (0.75 ml). After 2.5 h, additional tetrabutylammonium fluoride in THF (1 M; 0.08 mL, 0.08 mmol) was added and the reaction mixture was stirred for an additional 18 h. The reaction mixture was diluted with DCM and washed sequentially with saturated aqueous NaHCO ₃ and saturated aqueous sodium chloride solution, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica with a gradient elution of 0.5-10% MeOH in DCM. The product fractions were concentrated under reduced pressure and the resulting residue was further purified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5 μm silica, 19 mm in diameter, 150 mm in length) using an elution gradient of 45 to 85% acetonitrile in ( water containing 0.2% NH4OH). Product fractions were concentrated under reduced pressure to give the first eluted isomer 1 (2.7 mg, 7%) and the second eluted isomer (2.1 mg, 5%) 3 - ((6S, 8R) -6- (2.6 -difluoro-4- (1- (3-fluoropropyl) azetidin3-ylamino) phenyl) -8-methyl-8,9-dihydro-3H-pyrazolo [4,3-f] isoquinoline-7 (6H) -yl) - 2-fluoro-2-methylpropan1-ol in the form of pale yellow films.

Isomer 1: ¹ H NMR (500 MHz, CD ₂ Cl ₂ , 27 ° C) 1.13 (3H, d), 1.17 (3H, d), 1.66-1.78 (3H, m), 2.54 (2H, t), 2.69 (1H, dd), 2.81-2.94 (3H, m), 3.00 (1H, dd), 3.31 (1H, dd), 3 , 51-3.59 (1H, m), 3.60-3.71 (4H, m), 3.95-4.03 (1H, m), 4.38 (1H, br s), 4, 46 (2H, dt), 5.26 (1H, s), 6.01 (2H, br d), 6.84 (1H, d), 7.21 (1H, d), 8.03 (1H, s), 10.19 (1H, br s);

mass / charge: ES ⁺ [M + H] ⁺ = 520.

Isomer 2: 1H NMR (500 MHz, CD ₂ Cl ₂ , 27 ° C) 1.05 (3H, d), 1.09 (3H, d), 1.65-1.80 (2H, m), 2 , 54 (2H, t), 2.62 (1H, dd), 2.83-2.88 (2H, m), 2.91-3.00 (1H, m), 3.13-3.24 (1H, m), 3.33-3.46 (2H, m), 3.48-3.58 (1H, m), 3.66 (2H, q), 3.96-4.04 (2H , m), 4.40 (1H, dt), 4.41-4.53 (2H, m), 4.64 (1H, br d), 5.03 (1H, s), 6.06 (2H , br d), 6.76 (1H, d), 7.19 (1H, d), 8.03 (1H, s), 10.19 (1H, br s);

mass / charge: ES ⁺ [M + H] ⁺ 520.

Procedures used to prepare N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2fluoro-2-methylpropyl) -8-methyl-6,7,8,9- tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3-fluoropropyl) azetidin-3-amine starting material are described below.

Obtaining dimethyl 2-fluoro-2-methylpropanedioate o o

Sodium hydride (60% dispersion in mineral oil; 2.93 g, 73.3 mmol) was added to a solution of dimethyl 2-fluoromalonate (10.0 g, 66.6 mmol) in THF (218 mL) with vigorous stirring. After 30 min, iodomethane (4.56 ml, 73.3 mmol) was added. The reaction mixture was stirred for an additional 3 hours. The reaction mixture was quenched with water and then extracted with EtOAc (4x100 ml).

- 110 037533

The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give dimethyl 2-fluoro-2-methylpropanedioate (8.4 g, 77%) as an orange oil.

¹ H NMR (300 MHz, DMSO-d6, 27 ° C) 1.72 (3H, d), 3.77 (6H, s).

Obtaining 2-fluoro-2-methylpropane-1,3-diol

Lithium aluminum hydride (4.50 g, 112.6 mmol) was added in portions to a stirred solution of dimethyl 2-fluoro-2-methylpropanedioate (8.40 g, 51.2 mmol) in THF (205 ml) at 0 ° C. The reaction mixture was warmed to room temperature and stirred under these conditions for 1 hour. The reaction mixture was then cooled to 0 ° C and carefully quenched by sequential dropwise addition of water (5.85 ml), an aqueous solution containing 15 wt.% NaOH (5.85 ml), and water (18 ml). The resulting gelatinous suspension was rapidly stirred for 1 hour. The precipitate was removed by filtration and the filtrate was concentrated under reduced pressure. The resulting residue was dissolved in CHCl ₃ / IPA (3: 1), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 2-fluoro-2-methylpropane-1,3-diol (3.28 g, 46 %) as an orange oil. This oil was used in the next step without further purification.

¹ H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.18 (3H, d), 3.42 (4H, dd), 4.80 (2H, t).

Preparation of 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropan-1-ol

Sodium hydride (60% dispersion in mineral oil; 1.1 g, 27.5 mmol) was added to a stirred solution of 2-fluoro-2-methylpropane-1,3-diol (2.7 g, 25 mmol) in THF (93 ml) at 0 ° C and the reaction mixture was stirred for 1 h. Tert-butylchlorodiphenylsilane (6.5 ml, 25 mmol) was added and the reaction mixture was stirred for an additional 1 h. The reaction mixture was quenched with water, the layers were separated and the aqueous layer was extracted with using EtOAc (2x100 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and adsorbed onto silica gel under reduced pressure. Purification by flash chromatography on silica, gradient elution from 0 to 50% EtOAc in hexanes, afforded 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropan-1-ol (4.4 g , 51%) as a transparent resin.

1H NMR (300 MHz, DMSO-d6, 27 ° C) 1.00 (9H, s), 1.27 (3H, d), 3.48 (1H, dd), 3.53 (1H, dd), 3.66 (1H, d), 3.73 (1H, br s), 4.93 (1H, t), 7.40-7.48 (6H, m), 7.59-7.66 (4H , m);

mass / charge: ES ⁺ [M + H] ⁺ 347.

Preparation of 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyltrifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (1.3 ml, 7.6 mmol) was added dropwise to a stirred solution of 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropan-1-ol (2.2 g, 6, 35 mmol) and 2,6-dimethylpyridine (1.28 ml, 7.6 mmol) in DCM (22 ml) at -10 ° C (salt / ice bath). The reaction mixture was maintained under these conditions for 1.5 hours. The reaction mixture was then diluted with DCM (100 ml) and washed sequentially with aqueous HCl (1N), saturated aqueous NaHCO3 and saturated aqueous NaCl. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl trifluoromethanesulfonate (3.2 g) as a red oil. This oil was used in the next step without further purification.

Preparation of a diastereoisomeric mixture of 3 - ((2R) -2 - ((3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2methylpropyl) amino) propyl) -2-methylaniline

- ((tert-Butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyltrifluoromethanesulfonate (3.04 g,

6.35 mmol) was added to a solution of (R) -3- (2-aminopropyl) -2-methylaniline (1.04 g, 6.35 mmol) and diisopropylethylamine (1.65 ml, 9.53 mmol) in 1, 4-dioxane (24 ml). The reaction mixture was heated at 85 ° C for 18 h. After cooling, the reaction mixture was diluted with DCM (250 ml) and washed with water. The aqueous layer was extracted with DCM (2x100 ml) and the combined organics were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to

- 111 037533

25% MeOH in DCM. The product fractions were concentrated under reduced pressure to give 3 ((2R) -2 - ((3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) amino) propyl) -2-methylaniline (2.10 d, 95%) as a resin.

1H NMR (300 MHz, DMSO-d6, 27 ° C) 0.90 (3H, dd), 1.01 (9H, d), 1.29 (3H, dd), 1.41 (1H, br s) , 1.97 (3H, d), 2.29-2.43 (1H, m), 2.65-2.91 (4H, m), 3.62-3.82 (2H, m), 4 , 67 (2H, s), 6.28-6.36 (1H, m), 6.47 (1H, dd), 6.69-6.81 (1H, m), 7.40-7.52 (6H, m), 7.61-7.69 (4H, m);

mass / charge: ES ⁺ [M + H] ⁺ = 494.

Obtaining a diastereoisomeric mixture (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -3,5-dimethyl- 1,2,3,4-tetrahydroisoquinoline-6-amine Br

Δ ^H 2 ^N ---

4-Bromo-2,6-difluorobenzaldehyde (628 mg, 2.84 mmol) was added to a stirred solution 3 - ((2R) -2 - ((3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2- methylpropyl) amino) propyl) -2-methylaniline (700 mg, 1.42 mmol) and water (0.128 ml, 7.10 mmol) in a mixture of acetic acid (10 ml) and toluene (4 ml). The reaction mixture was heated at 90 ° C for 18 hours before heating under reflux for an additional 4 hours. The reaction mixture was allowed to cool and concentrated under reduced pressure. The resulting residue was dissolved in DCM and washed with saturated aqueous NaHCO ₃ solution. The aqueous phase was extracted with DCM (20 ml) and the combined organic layers were concentrated under reduced pressure. The residue was dissolved in MeOH / DCM (5: 1, 18 ml) and then hydroxylamine hydrochloride (148 mg, 2.13 mmol) and sodium acetate (233 mg, 2.84 mmol) were added. The mixture was stirred at 35 ° C for 5 min and then concentrated under reduced pressure. The resulting residue was dissolved in EtOAc and washed sequentially with saturated aqueous NaHCO ₃ and saturated aqueous NaCl before being dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 35% EtOAc in hexanes, to give (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (3 - ((tert -butyl diphenylsilyl) oxy) -2-fluoro-2-methylpropyl) 3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (339 mg, 34%) as a pale yellow film.

¹ H NMR (500 MHz, CDCl3, 27 ° С) 1.05 (9H, d), 1.19 (3H, dd), 2.08 (3H, s), 2.35 (1H, dd), 2 , 47-2.67 (2H, m), 2.84-3.01 (1H, m), 3.02-3.14 (1H, m), 3.22 (1H, dd), 3.40 -3.63 (3H, m), 3.66-3.79 (1H, m), 3.92 (1H, dd), 5.15 (1H, d), 6.38-6.47 (2H , m), 6.83 (1H, d), 6.93-7.00 (1H, m), 7.38-7.49 (6H, m), 7.59-7.71 (4H, m ), 8.69 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ 695.

Preparation of a diastereoisomeric mixture (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-6, 7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline

Sodium nitrite (33.6 mg, 0.49 mmol) as a solution in water (0.400 ml) was added dropwise to a cooled solution of (1S, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2methylpropyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine (339 mg, 0.49 mmol) in propionic acid (4 ml) at -20 ° C and the reaction mixture was stirred for 30 min under these conditions. Ice-cold EtOAc (20 ml) was added followed by portionwise addition of saturated aqueous NaHCO ₃ (5 ml). The biphasic mixture was vigorously stirred and neutralized by the slow addition of solid Na ₂ CO ₃ . The phases were separated and the organic layers were washed with saturated aqueous NaHCO ₃ (2x30 ml) and saturated aqueous NaCl (30 ml) before being dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 40% EtOAc in hexanes, to give (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3- ( (tert-butyldiphenylsilyl) oxy) -2fluoro-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (206 mg, 60%) as pale -yellow solid.

1H NMR (500 MHz, CDCl3, 27 ° С) 0.98 (4.5H, s), 1.00 (1.5H, d), 1.02 (1.5H, d), 1.04 (4 , 5H, s), 1.15 (1.5H, d), 1.23 (1.5H, d), 2.30-2.47 (0.5H, m), 2.60 (0.5H , dd), 2.81 (0.5H, dd), 2.89 (0.5H, dd), 3.02 (0.5H, dd), 3.063.21 (1H, m), 3.22- 3.33 (0.5H, m), 3.37-3.51 (1H, m), 3.51-3.60 (0.5H, m), 3.61-3.72 (0.5H , m), 3.73-3.83

- 112 037533 (0.5H, m), 3.89 (0.5H, dd), 5.22 (0.5H, s), 5.27 (0.5H, s), 6.70 (1H, dd), 6.81 (1H, br d), 6.91-7.00 (1H, m), 7.14 (1H, t), 7.33-7.47 (6H, m), 7, 55-7.67 (4H, m), 8.07 (1H, br d). NH indazole was not observed, mass / charge: ES + [M + H] ⁺ = 706.

Preparation of a diastereoisomeric mixture (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9 tetrahydro-3H-pyrazolo [4,3-f] isoquinoline Br

3,4-Dihydro-2H-pyran (0.129 ml, 1.41 mmol) and p-toluenesulfonic acid monohydrate (2.7 mg, 0.01 mmol) were added to a stirred solution of (6S, 8R) -6- (4- bromo-2,6-difluorophenyl) -7- (3 ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrαzolo [4 , 3f] isoquinoline (200 mg, 0.28 mmol) in DCM (2 ml). The reaction mixture was heated at 45 ° C for 21 hours. The reaction mixture was allowed to cool and then it was diluted with DCM, washed with saturated aqueous NaHCO ₃ solution, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 5% to 30% EtOAc in hexanes, to give (6S, 8R) -6- (4bromo-2,6-difluorophenyl) -7- (3 - ((tert -butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3- f] isoquinoline (176 mg, 79%) as a colorless film.

Mass / charge: ES ⁺ [M + H] ⁺ = 790.

Preparation of a diastereoisomeric mixture of tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-3- ( tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) amino) azetidine-1- carboxylate

The vial was loaded with a magnetic anchor, (6S, 8R) -6- (4-bromo-2,6-difluorophenyl) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl -3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (126 mg, 0.16 mmol), tert-butyl-3- aminoazetidine-1carboxylate (41 mg, 0.24 mmol), Pd ₂ dba ₃ (9.3 mg, 0.01 mmol), 4,5-bis- (diphenylphosphino) -9.9 dimethylxanthene (Xantphos) (16 mg, 0 , 02 mmol) and cesium carbonate (156 mg, 0.48 mmol). The vial was sealed, evacuated and filled with nitrogen (3x) before adding 1,4-dioxane (1 ml) using a syringe. The mixture was stirred at ambient temperature for 2 min and then heated at 90 ° C for 16 h. The mixture was allowed to cool and then diluted with EtOAc, filtered through diatomaceous earth and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 40% EtOAc in hexanes, to give tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert- butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3- f] isoquinolin-6-yl) -3,5-difluorophenyl) amino) azetidine-1-carboxylate (95 mg, 68%) as a pale yellow solid.

Mass / charge: ES ⁺ [M + H] ⁺ = 883.

Obtaining a diastereoisomeric mixture of N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro- 3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) azetidin-3-amine

- 113 037533

Formic acid (0.5 ml, 13.04 mmol) was added to tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2- methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3, 5-difluorophenyl) amino) azetidine-1-carboxylate (53 mg, 0.06 mmol) and the resulting solution was stirred at ambient temperature for 18 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was dissolved in THF (0.5 ml ) and treated with an aqueous solution of NaOH (5N; 0.081 ml, 2.40 mmol). The mixture was stirred at 30 ° C for 20 h and then maintained at 40 ° C for 4 h. Additional aqueous NaOH solution (5N; 0.081 mL, 2.40 mmol) was added and the reaction mixture was stirred at 40 ° C for 46 h before heating at 55 ° C for 5 h. Allowed to cool the reaction mixture to ambient temperature. At the same time, HCl in dioxane (4N; 0.38 ml, 1.5 mmol) was added to a separate vial to a stirred solution of tert-butyl-3 - ((4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4 , 3-f] isoquinolin-6-yl) -3,5-difluorophenyl) amino) azetidine-1-carboxylate (89 mg, 0.10 mmol) in MeOH (0.5 ml). The reaction mixture was stirred at room temperature for 5 hours and then concentrated under reduced pressure. The resulting residue was combined with the previous reaction mixture (as a mixture in aqueous NaOH (5N) and THF). The new mixture was diluted with DCM (5 ml), the phases were separated and the aqueous phase was extracted with DCM (2x5 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure to give N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) azetidin-3-amine (177 mg) as pale yellow solid which was used in the next step without further purification.

Mass / charge: ES + [M + H] ⁺ = 698.

Obtaining a diastereoisomeric mixture of N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro- 3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3fluoropropyl) azetidin-3-amine

1-Fluoro-3-iodopropane (0.026 ml, 0.16 mmol) was added to a stirred solution of N- (4 - ((6S, 8R) 7- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2 -methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) azetidin-3-amine (115 mg, 0.16 mmol) and diisopropylethylamine (0.058 ml, 0.33 mmol) in NMP (0.75 ml). The reaction mixture was stirred for 16 h at room temperature before being diluted with EtOAc and washed with saturated aqueous NaHCO ₃ solution. The layers were separated and the aqueous layer was extracted with EtOAc. The combined organic layers were washed with saturated aqueous NaCl solution (2x5 ml), dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 1 to 20% MeOH in DCM, to give N- (4 - ((6S, 8R) -7- (3 - ((tert-butyldiphenylsilyl) oxy) - 2-fluoro-2-methylpropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3,5-difluorophenyl) -1- (3- fluoropropyl) azetidine-3-amine (82 mg, 66%) as a pale yellow film.

Mass / charge: ES ⁺ [M + H] ⁺ = 758.

Example 25.

Preparation of 6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -N - (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

HCl in dioxane (4N; 0.86 ml, 3.4 mmol) was added dropwise to a solution of tert-butyl-3 - ((6 - ((6S, 8R) 7- (2,2-difluoroethyl) -8- methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) pyridin-3-yl) amino) azetidine- 1-carboxylate (0.20 g, 0.34 mmol) in MeOH (2.5 ml) and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure to give N- (azetidin-3-yl ) -6 - ((6S, 8R) -7- (2,2-difluoroethyl) - 114 037533

8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyrudin-3-amine hydrochloride (200 mg) as a solid.

Mass / charge: (ES +), [M + H] ⁺ = 399.

DMF (0.5 ml) and 1-fluoro-3-iodopropane (0.036 ml, 0.34 mmol) were added sequentially at ambient temperature. An excess of diisopropylethylamine (1.2 ml, 6.80 mmol) was then added dropwise. The reaction mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure. The resulting residue was purified by a C18 reverse phase flash chromatography column, elution gradient from 20% to 75% acetonitrile in (water containing 0.2% NH4OH) to give 6 - ((6S, 8R) -7- (2, 2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine (45 mg, 29%) as a solid.

Ή NMR (500 MHz, DMSO-d6, 27 ° С) 1.04 (3H, d), 1.60-1.75 (2H, m), 2.51-2.70 (3H, m), 2 , 79-2.96 (3H, m), 3.03 (1H, dd), 3.13 (1H, br dd), 3.41-3.53 (1H, m), 3.74 (2H, br s), 3.98 (1H, br d), 4.45 (2H, dt), 4.86 (1H, s), 5.83 (1H, tt), 6.26 (1H, d), 6.74 (1H, d), 6.81 (1H, dd), 6.96 (1H, d), 7.18 (1H, d), 7.76 (1H, d), 8.04 (1H , s), 12.95 (1H, s);

mass / charge: (ES ⁺ ) [M + H] ⁺ = 459.

Procedures used to prepare tert-butyl-3 - ((6 - ((6S, SR) -7- (2,2-difluoroethyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3yl) amino) azetidine-1-carboxylate starting material are described below.

Preparation of (1 S, 3R) -1 - (5-bromopyridin-2-yl) -2- (2,2-difluoroethyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine

Bg

5-Bromopicolinaldehyde (2.74 g, 14.7 mmol) was added to a solution of (R) -3- (2 - ((2,2 difluoroethyl) amino) propyl) -2-methylaniline (1.6 g, 7.0 mmol ) in acetic acid (34.4 ml) and water (0.631 ml, 35.0 mmol). The reaction mixture was heated at 80 ° C for 3 h and then concentrated under reduced pressure. The resulting residue was dissolved in MeOH (40 ml) and sodium acetate (1.15 g, 14.0 mmol) and hydroxylamine hydrochloride (0.730 g, 10.5 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure. The residue was dissolved in water, neutralized with saturated aqueous NaHCO ₃ and then extracted with EtOAc. The combined organic layers were concentrated under reduced pressure and the resulting residue was purified by flash silica chromatography, elution gradient from 0 to 90% EtOAc in hexanes. The product fractions were concentrated under reduced pressure and the resulting residue was further purified by flash chromatography on silica, elution gradient from 0 to 90% EtOAc in hexanes, to give (1S, 3R) -1- (5-bromopyridin-2-yl) -2 - (2,2-difluoroethyl) -3,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-amine (1.40 g, 49%) as gum.

¹ H NMR (300 MHz, DMSO-d6, 27 ° C) 0.99 (3H, d), 1.93 (3H, s), 2.38-2.47 (1H, m), 2.51- 2.62 (1H, m), 2.72 (1H, dd), 2.92-3.14 (1H, m), 3.23-3.29 (1H, m), 4.65 (2H, s), 4.79 (1H, s), 5.98 (1H, tt), 6.39 (2H, s), 7.22 (1H, d), 7.91 (1H, dd), 8, 55 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 396.

Obtaining (6S, 8R) -6- (5-bromopyrudin-2-yl) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f ] isoquinoline

Br

Acetic acid (541 mg, 9.01 mmol) was added to a stirred solution of (1S, 3R) -1- (5-bromopyridin-2-yl) -2- (2,2-d ifluoroethyl) -3,5-dimethyl-1, 2,3,4-tetrahydroisoquinolin-6-amine (714 mg, 1.80 mmol) in CHCl ₃ (8 ml). The reaction mixture was cooled to 0 ° C and a solution of isopentyl nitrite (422 mg, 3.60 mmol) in CHCl ₃ (1 ml) was added dropwise. The reaction mixture was stirred at 0 ° C for 2 h and then quenched by the slow addition of a solution of NaHCO3 (1.5 g, 18 mmol) in water (20 ml). The phases were separated and the organic layer was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 5% to 35% EtOAc in hexanes, to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoroethyl ) -8-methyl-6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (438 mg, 60%) as a light brown solid.

- 115 037533

1H NMR (300 MHz, CDC13, 27 ° C) 1.13 (3H, d), 2.68-2.85 (1H, m), 2.91 (1H, dd), 2.98-3.17 (1H, m), 3.34 (1H, dd), 3.47-3.63 (1H, m), 5.05 (1H, s), 5.63 (1H, tt), 6.87 ( 1H, d), 7.19 (1H, d), 7.28 (1H, d), 7.71 (1H, dd),

8.04 (1H, d), 8.57 (1H, dd). NH indazole was not observed, mass / charge: ES ⁺ [M + H] ⁺ = 405.

Obtaining (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2yl) -6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

Br А Г ^ ° - r ^N

Ν ^ / \ - (/ g f

3,4-Dihydro-2H-pyran (0.294 ml, 3.23 mmol) was added to a solution of (6S, 8R) -6- (5-bromopyridine-2yl) -7- (2,2-difluoroethyl) -8-methyl -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (438 mg, 1.08 mmol) and p-toluenesulfonic acid monohydrate (21 mg, 0.11 mmol) in DCM (4 ml). The reaction mixture was heated at 100 ° C for 6 h under microwave conditions (300 W). The reaction mixture was then allowed to cool and then washed with saturated aqueous NaHCO ₃ , dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 10 to 30% EtOAc in hexanes to give (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2 difluoroethyl) - 8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (441 mg, 83%) as light brown gummy solid.

Mass / charge: ES ⁺ [M + H] ⁺ = 491.

Preparation of tert-butyl-3 - ((6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 6,7,8 , 9-tetrahydro-3H-pyrαzolo [4,3-f] isoquinolin-6-yl) pyridin-3-yl) amino) azetidine-1-carboxylate

The vial was loaded with a magnetic anchor, (6S, 8R) -6- (5-bromopyridin-2-yl) -7- (2,2-difluoroethyl) -8methyl-3- (tetrahydro-2H-pyran-2-yl) - 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (0.22 g, 0.44 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (0.151 g, 0, 88 mmol), cesium carbonate (0.29 g, 0.88 mmol) and BrettPhos 3rd generation pre-catalyst (0.040 g, 0.040 mmol). The vial was sealed, evacuated and filled with nitrogen. 1,4-dioxane (4 ml) was added and the vial was evacuated again and filled with nitrogen. The reaction mixture was stirred at 110 ° C for 13 hours and then concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 10 to 30% EtOAc in hexanes, to give tert-butyl-3 - ((6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8 -methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-yl) amino ) α-zetidine-1-carboxylate (0.20 g, 78%) as a beige solid.

1H NMR (300 MHz, CD3OD, 27 ° C) 1.10 (3H, dd), 1.43 (9H, s), 1.56-1.72 (2H, m), 1.72-1.87 (1H, m), 1.92-1.99 (1H, m), 2.03-2.17 (1H, m), 2.38-2.53 (1H, m), 2.62-2 , 82 (1H, m), 2.88-3.14 (2H, m), 3.33-3.44 (1H, m), 3.49-3.60 (1H, m), 3.67 -3.83 (3H, m), 3.92-4.02 (1H, m), 4.17-4.33 (3H, m), 4.91 (1H, s), 5.34-5 , 57 (1H, m), 5.69-5.77 (1H, m), 6.79 (1H, d), 6.89 (1H, dd), 7.05 (1H, dd), 7, 34 (1H, d), 7.78-7.81 (1H, m), 8.07 (1H, s), no aniline NH observed, w / charge: ES ⁺ [M + H] ⁺ = 583.

Example 26.

Preparation of (6S, 8R) -7- (2,2-difluoroethyl) -6- (3 - ((1- (3-fluoropropyl) αzetidin-3-yl) oxy) pyridin-2-yl) 8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

HCl in dioxane (4N; 0.97 ml, 3.9 mmol) was added dropwise to a solution of tert-butyl-3 - ((6 - ((6S, 8R) 7- (2,2-difluoroethyl) -8- methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyranazolo [4,3

- 116 037533

f] isoquinolin-6-yl) pyridin-3-yl) oxy) azetidine-1-carboxylate (0.227 g, 0.39 mmol) in MeOH (3 ml) and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture concentrated under reduced pressure to give (6S, 8R) -6- (5- (azetidin-3-yloxy) pyridin-2-yl) 7- (2,2-difluoroethyl) -8-methyl-6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline hydrochloride (0.235 g; HCl equivalents not determined) as a solid which was used without further purification.

Mass / charge: (ES +), [M + H] ⁺ = 400.

1-Fluoro-3-iodopropane (0.041 ml, 0.34 mmol) was added to a stirred solution of (6S, 8R) -6- (5 (azetidin-3-yloxy) pyrudin-2-yl) -7- (2.2 -difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline hydrochloride (235 mg) in DMF (0.6 ml) at ambient temperature. An excess of diisopropylethylamine (1.36 mL, 7.80 mmol) was added dropwise. The reaction mixture was stirred for 18 hours at room temperature and then concentrated under reduced pressure. The resulting residue was purified using a C18 reverse phase flash chromatography column eluted from 20% to 75% acetonitrile in (water containing 0.2% NH4OH). The product fractions were combined and lyophilized to give (6S, 8R) -7- (2,2-difluoroethyl) -6- (5 - ((1- (3-fluoropropyl) azetidin-3yl) oxy) pyridin-2-yl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (53 mg, 30%) as a solid.

¹ H NMR (500 MHz, DMSO-d6, 27 ° C) 1.05 (3H, d), 1.60-1.77 (2H, m), 2.54-2.70 (3H, m), 2.85 (1H, dd), 2.97-3.22 (4H, m), 3.39-3.50 (1H, m), 3.87 (2H, br s), 4.45 (2H , dt), 4.80-4.92 (1H, m), 5.00 (1H, s), 5.93 (1H, tt), 6.78 (1H, d), 7.19-7, 26 (3H, m), 8.05 (1H, s), 8.06 (1H, d), 12.97 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 460.

Procedures used to prepare tert-butyl-3 - ((6 - ((6,6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-3 (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3yl) oxy) azetidine-1-carboxylate starting material are described below.

Preparation of tert-butyl-3 - ((6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl) 6,7,8 , 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-yl) oxy) azetidine-1-carboxylate n ' ^Boc tert-Butyl 3-hydroxyazetidine-1-carboxylate (227 mg, 1.31 mmol), (6S, 8R) -6- (5-bromopyridin-2yl) -7- (2,2-difluoroethyl) -8-methyl-3- (tetrahydro-2H-pyran-2-yl ) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (215 mg, 0.44 mmol), ^{RockPhos 3rd} generation precatalyst (37.1 mg, 0.04 mmol) and cesium carbonate (285 mg, 0.88 mmol) was suspended in toluene (4 ml) in a sealed microwave vial. The reaction mixture was heated at 110 ° C for 1 h under microwave conditions (300 W). The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by flash chromatography on silica eluting with 20 to 60% EtOAc in hexanes to give tert-butyl 3 - ((6 - ((6S, 8R) -7- (2,2-difluoroethyl) 8-methyl-3- (tetrahydro-2H-pyran-2-yl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl ) pyridin3-yl) oxy) azetidine-1-carboxylate (227 mg, 89%) as a solid.

Ή NMR (300 MHz, CD3OD, 27 ° С) 1.11 (3H, dd), 1.41-1.43 (9H, m), 1.61-1.66 (1H, m), 1.66 -1.86 (2H, m), 2.04-2.17 (1H, m), 2.36-2.54 (1H, m), 2.59-2.79 (1H, m), 2 , 89-3.02 (1H, m), 3.03-3.17 (1H, m), 3.31-3.43 (1H, m), 3.48-3.60 (1H, m) , 3.70 (2H, dd), 3.88-3.95 (2H, m), 4.09-4.13 (1H, m), 4.27-4.39 (2H, m), 4 , 99-5.08 (2H, m), 5.64 (1H, tt), 5.73 (1H, dt), 6.81 (1H, dd), 7.20 (1H, dd), 7, 27 (1H, dd), 7.36 (1H, d), 8.04-8.10 (2H, m);

mass / charge: ES ⁺ [M + H] ⁺ = 584.

Example 27.

1- (3-Fluoropropyl) -N- (4 - ((6R, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [ 4,3] isoquinolin-6-yl) phenyl) azetidin-3-amine sodium tert-butoxide (2.118 g, 22.06 mmol) and BrettPhos G3 (0.166 g, 0.18 mmol) were added to a degassed solution of 1- (3 -fluoropropyl) azetidin-3-amine (0.632 g, 4.78 mmol) and (6R, 8R) -6- (4

- 117 037533 bromophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (1.56 g, 3 , 68 mmol) in 1,4-dioxane (18.4 ml) and the reaction mixture was heated to 90 ° C and stirred overnight. After cooling, the reaction mixture was diluted with EtOAc (20 ml) and water (20 ml) and the layers were separated. The aqueous layer was extracted with EtOAc (20 ml), then the combined organics were dried and evaporated. Purification was carried out by HPLC (Waters CSH C18 OBD column, 5 μm silica, 30 mm in diameter, 100 mm in length) using mixtures of water (containing 0.1% NH ₃ and MeCN as eluents) with decreasing polarity to obtain a resin ... This was taken up in methanol (5 ml), then water (95 ml) was added and the mixture was suspended overnight at room temperature. The resulting solid was collected by filtration, washed with 5% methanol in water and dried in a vacuum oven at 50 ° C overnight to give 1- (3-fluoropropyl) -N- (4 - ((6R, 8R) -8 -methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (0.935 g, 54%) as a colorless solid.

¹ H NMR (500 MHz, CDCl3, 27 ° C) 1.13 (3H, d), 1.68-1.83 (2H, m), 2.59 (2H, t), 2.77 (1H, dd), 2.86 (2H, t), 2.89-2.99 (1H, m), 3.04 (1H, dd), 3.15-3.26 (1H, m), 3.37 -3.48 (1H, m), 3.71-3.76 (2H, m), 3.93 (1H, d), 4.10 (1H, q), 4.43 (1H, t), 4.53 (1H, t), 4.99 (1H, s), 6.41-6.46 (2H, m), 6.97 (1H, d), 7.00 (2H, d), 7 , 26 (1H, s), 8.06 (1H, d), 10.10 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 476.

(6R, 8R) -6- (4-Bromophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline, used as a starting material was synthesized as follows.

(6R, 8R) -6- (4-Bromophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline

Trifluoroacetic acid (1.85 ml) was added to a solution of (R) -1- (1H-indazol-4-yl) -N- (2,2,2trifluoroethyl) propan-2-amine (2.0 g, 7.77 mmol) and 4-bromobenzaldehyde (7.19 g, 38.9 mmol) in toluene (37 ml) and the resulting mixture was stirred at 90 ° C. for 30 hours. The reaction mixture was cooled and partitioned between DCM (100 ml) and saturated aqueous sodium bicarbonate solution (50 ml). The layers were separated and the organic layer was concentrated in vacuo. Purification was carried out by column chromatography on silica gel eluting with 0-50% ethyl acetate in heptane to give (6R, 8R) -6- (4-bromophenyl) -8-methyl-7- (2,2,2-trifluoroethyl) - 6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f] isoquinoline (1.560 g, 47%) as a foam and in a 6: 1 isomer ratio.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 2.80 (1H, dd), 2.94 (1H, dd), 3.05 (1H, dd), 3.25 (1H, dd), 3.33 (1H, ddd), 5.04 (1H, s), 6.95 (1H, d), 7.08-7.12 (2H, m), 7 , 28-7.33 (1H, m), 7.37-7.40 (2H, m), 8.08 (1H, d), 10.12 (1H, s);

mass / charge: ES ^- [M-H] ^- = 422.

Example 28.

Preparation of 1- (3-fluoropropyl) -N- (3-methoxy-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H -pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine

BrettPhos 3rd generation precatalyst (10 mg, 0.01 mmol) and sodium tert-butoxide (0.127 g, 1.32 mmol) were added in one portion to a degassed solution of 1- (3-fluoropropyl) azetidine-3-amine (0.033 g , 0.25 mmol) and (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H -pyrazolo [4,3-f] isoquinoline (0.10 g, 0.22 mmol) in 1,4-dioxane (1.10 ml). The orange mixture was then immersed in an oil bath that had been preheated to 50 ° C. After 5 min, the reaction mixture was cooled to room temperature. In a separate flask, sodium tert-butoxide (1.81 g, 18.8 mmol) and the 3rd generation BrettPhos precatalyst (0.17 g, 0.19 mmol) were added to the degassed solution of (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (1.71 g , 3.76 mmol) and 1- (3-fluoropropyl) azetidin-3-amine

- 118 037533 (0.597 g, 4.52 mmol) in 1,4-dioxane (18.8 ml). The light orange mixture was immersed in an oil bath that had been preheated to 50 ° C. After 5 min, the reaction mixture was cooled to room temperature. Once cooled, the reaction mixtures were combined, diluted with ethyl acetate and washed sequentially with water (x2) and saturated aqueous sodium chloride solution. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting orange oil was purified by flash chromatography on silica eluting with 0-10% methanol in DCM to give 1- (3-fluoropropyl) -N- (3-methoxy-4 - ((6S, 8R) 8- methyl 7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3 _: /] isoquinolin-6-yl) phenyl) azetidine-3amine (1.84 g , 92%) as a solid and a mixture of trans and cis isomers ~ 84: 16, based on the UV HPLC profile. This material was isolated using preparative SFC (column: Chiralpak AD, 21.2x250 mm, 5 μm; 75 ml / min.) Eluting with 20% (methanol containing 0.2% NH ₄ OH) in CO ₂ to give 1- (3-fluoropropyl) -N- (3-methoxy-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H- pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (1.01 g) as a light orange solid and its corresponding second eluting peak. This material was further purified by flash chromatography on silica, elution gradient from 0 to 30% methanol in ethyl acetate, to give 1- (3-fluoropropyl) -N- (3-methoxy-4 - ((6S, 8R) -8 -methyl-7 (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine (0.954 g ) as a white solid. This material was ultimately purified by preparative HPLC (column: Xbridge C18, 30x100 mm, 5 μm, 40 ml / min) eluting with 40 to 70% acetonitrile in (water containing 0.2% NH4OH). The product fractions were combined, washed with ethyl acetate (x3) and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure to give 1- (3-fluoropropyl) -N- (3-methoxy -4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl ) phenyl) azetidine-3-amine (551 mg, 27%) as an off-white foam solid.

¹ H NMR (500 MHz, DMSO-d6, 27 ° C) 1.04 (3H, d), 1.63 (2H, dtt), 2.44 (2H, t), 2.70 (2H, dd) , 2.81 (1H, dd), 2.84-2.94 (1H, m), 3.09 (1H, dd), 3.34 (1H, br s), 3.44 (1H, dqd) , 3.61 (2H, dd), 3.77 (3H, s), 3.90 (1H, m), 4.43 (2H, dt), 5.28 (1H, s), 5.87 ( 1H, dd), 6.02 (1H, d), 6.16 (1H, d), 6.32 (1H, d), 6.65 (1H, d), 7.18 (1H, d), 8.02 (1H, s); 12.95 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 506.

Procedures used to prepare (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline starting material are described below.

Preparation of (R) -N- (1- (3-amino-2-methylphenyl) propan-2-yl) -2,2,2-trifluoroacetamide

Ethyl 2,2,2-trifluoroacetate (3.75 ml, 31.5 mmol) was added to a dark amber-red solution of (R) -3- (2-aminopropyl) -2-methylaniline (5.17 g, 31, 5 mmol) and triethylamine (4.83 ml, 34.6 mmol) in MeOH (70.1 ml). After 15 minutes, the reaction mixture was concentrated to a dark amber oil. The oil was dissolved in ethyl acetate, washed with water and the aqueous layer was extracted with ethyl acetate (x2). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting oil was purified by flash chromatography on silica, elution gradient from 0 to 40% ethyl acetate in hexanes, to give (R) -N- (1- (3-amino-2methylphenyl) propan-2-yl) -2.2 , 2-trifluoroacetamide (6.47 g, 79%) as a yellow-orange oil which crystallized to a light orange solid upon standing.

¹ H NMR (300 MHz, DMSO-06, 27 ° С) 1.11 (3H, d), 2.01 (3H, s), 2.63 (1H, dd), 2.78 (1H, dd) , 3.93-4.05 (1H, m), 4.71 (2H, s), 6.36 (1H, dd), 6.49 (1H, dd), 6.78 (1H, t), 9.25 (1H, br d);

mass / charge: ES ⁺ [M + H] ⁺ = 261.

Preparation of (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline

Borane-tetrahydrofuran complex in THF (1 M; 149 ml, 149 mmol) was added using a syringe (6x25 ml) to a solution of (R) -N- (1- (3-amino-2-methylphenyl) propan-2-yl) -2,2,2-trifluoroacetamide (6.47 g, 24.9 mmol) in tetrahydrofuran (81 ml) at 0 ° C. The ice bath was removed and gas evolution was observed upon heating. As soon as further gas evolution ceased to be visually observed (~ 20 min), the reaction mixture was heated to 65 ° C. After 4 h, the reaction mixture was cooled and maintained at room temperature for 18 h. The reaction mixture was then cooled to 0 ° C and quenched with

- 119 037533 dropwise addition of methanol (22 ml). The reaction mixture was warmed to room temperature and after gas evolution ceased, the reaction mixture was heated to 65 ° C. Then, after 14 hours, the reaction mixture was cooled to room temperature and stirred under such conditions for 3.5 days before being concentrated under reduced pressure. The resulting oil was purified by flash chromatography on silica, elution gradient from 0 to 70% ethyl acetate in hexanes, to give (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (5.62 g, 92%) as a light yellow oil.

'H NMR (300 MHz, DMSO-d6, 27 ° C) 0.90 (3H, d), 1.98 (3H, s), 2.09-2.20 (1H, m), 2.25- 2.40 (1H, m), 2.72-2.84 (2H, m), 3.17-3.29 (2H, m), 4.69 (2H, s), 6.36 (1H, dd), 6.49 (1H, dd), 6.78 (1H, t);

mass / charge: ES ⁺ [M + H] + = 247

Preparation of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1,2,3,4tetrahydroisoquinoline-6-amine

Br

F

4-Bromo-2-methoxybenzaldehyde (1.83 g, 8.53 mmol) and (R) -2-methyl-3- (2 - ((2,2,2-trifluoroethyl) amino) propyl) aniline (2, 00 g, 8.12 mmol) was added to a solution of water (0.7 ml, 41 mmol) and acetic acid (40 ml). The resulting light yellow solution was immersed in an oil bath which had been preheated to 65 ° C. and maintained under these conditions for 18 hours. The resulting dark amber solution was then concentrated under reduced pressure (water bath: 55 ° C.). The resulting residue was diluted with ethyl acetate and washed with a saturated aqueous sodium hydrogen carbonate solution until the pH of the aqueous layer was confirmed to be 8 using a pH litmus strip.

The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was dissolved in methanol (20 ml) and hydroxylamine hydrochloride (0.152 g, 2.19 mmol) and excess potassium carbonate was added. After 5 minutes, the mixture was diluted with ethyl acetate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 50% ethyl acetate in hexanes, to give (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1,2,3,4tetrahydroisoquinoline-6-amine as a dark amber oil with a trans to cis isomer ratio of 83:17 based on NMR integration. The oil was re-concentrated from DCM and dried in vacuo to a light amber foam solid (2.55 g, 71%). A small amount of this material (150 mg) was isolated using preparative SFC (column: (S, S) Whelk-Ol, 21.2x250 mm, 5 μm; 75 ml / min) eluting with 15% (methanol containing 0, 2% NH ₄ OH) in CO ₂ to give the more slowly eluted (1S, 3R) -1- (4-bromo-2methoxyphenyl) -3,5-dimethyl-2- (2,2,2-trifluoroethyl) -1, 2,3,4-tetrahydroisoquinolin-6-amine (101 mg) and more rapidly eluted (1R, 3R) -1- (4-bromo-2-methoxyphenyl) -3,5-dimethyl-2- (2,2, 2-trifluoroethyl) -1,2,3,4tetrahydroisoquinoline-6-amine (18 mg) as pale yellow foam solids.

(1S, 3R): 'H NMR (300 MHz, DMSO-d6, 27 ° C) 1.00 (3H, d), 1.95 (3H, s), 2.43 (1H, dd), 2, 64-2.85 (2H, m), 3.19-3.43 (2H, m), 3.86 (3H, s), 4.67 (2H, s), 5.18 (1H, s) , 6.29 (1H, d), 6.41 (1H, d), 6.60 (1H, d), 6.96 (1H, dd), 7.18 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 443.

(1R, 3R): 'H NMR (300 MHz, DMSO-d6, 27 ° C) 1.19 (3H, d), 1.94 (3H, s), 2.53-2.60 (1H, m ), 2.78 (1H, br dd), 2.98-3.12 (1H, br m), 3.16-3.36 (2H, m), 3.84 (3H, s), 4, 61 (2H, s), 5.31 (1H, s), 6.20 (1H, d), 6.33 (1H, d), 7.03-7.13 (2H, m), 7.19 (1H, d);

mass / charge: ES ⁺ [M + H] ⁺ 443.

Obtaining (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3 -f] isoquinoline

A solution of sodium nitrite (0.413 g, 5.98 mmol) in water (3.83 ml) was added dropwise over 5 min to a stirred solution of (1S, 3R) -1- (4-bromo-2-methoxyphenyl) -3, 5-dimethyl-2- (2,2,2trifluoroethyl) -1,2,3,4-tetrahydroisoquinoline-6-amine (2.55 g, 5.75 mmol; mixture of trans and cis isomers 83:17) in propionic acid (19.2 ml) maintained at -15 ° C using a cooling bath,

- 120 037533 consisting of ice, solid sodium chloride and a saturated aqueous solution of sodium chloride. After 20 min, the reaction mixture was diluted with toluene (100 ml), which was precooled to -70 ° C. The resulting light yellow mixture was vigorously stirred and the cooling bath was removed after 5 minutes. Upon reaching room temperature, the red reaction mixture was maintained under these conditions for 1.5 hours and then washed with water (x2). The combined aqueous layers were extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to approximately 20% original volume. This mixture was diluted with EtOAc (20 ml) and washed with saturated aqueous sodium bicarbonate solution. Then solid potassium carbonate was added until gas evolution ceased and the mixture was tested for basicity using a pH litmus strip. The layers were separated and the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting oil was purified by flash chromatography on silica eluting from 0 to 70% ethyl acetate in hexanes to give (6S, 8R) -6- (4-bromo-2-methoxyphenyl) -8-methyl7- (2.2 , 2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (1.96 g, 75%) as a light orange foam solid and a mixture of trans - and cis-isomers ~ 9: 1, based on integration at NMR.

1H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.06 (3H, d), 2.80-2.98 (2H, m), 3.15 (1H, br dd), 3.33 -3.53 (2H, m), 3.90 (3H, s), 5.40 (1H, s), 6.63 (1H, d), 6.67 (1H, d), 6.96 ( 1H, dd), 7.20-7.27 (2H, m), 8.08 (1H, s), 13.00 (1H, s);

mass / charge: ES + [M + H] + = 454.

Example 29.

Preparation of 2-fluoro-N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) 6,7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridine-3-amine

Sodium tert-butoxide (1.08 g, 11.3 mmol) and the 3rd generation BrettPhos precatalyst (0.16 g, 0.18 mmol) were added to the degassed solution of (6S, 8R) -6- (5-bromo- 6-fluoropyridin-2-yl) -8-methyl-7 (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (2.00 g , 4.51 mmol) and 1- (3-fluoropropyl) azetidin-3-amine (0.90 g, 5.41 mmol) in 1,4-dioxane (22.6 ml) at room temperature. The red-orange mixture was immersed in an oil bath preheated to 44 ° C. After 22 minutes, the orange mixture was removed from the heat source and poured into ethyl acetate and saturated aqueous sodium chloride solution and the layers were separated. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 30% methanol in ethyl acetate, to give 2-fluoro-N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S , 8R) -8-methyl-7 (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (1.58 g) as a light yellow foam solid and a mixture of trans and cis isomers in a ratio of approximately 84:16 based on NMR integration. This material was isolated using preparative SFC (column: Lux Cellulose-4, 21.2x250 mm, 5 μm; 70 ml / min) eluting with 35% (methanol containing 0.2% NH4OH) in CO ₂ to give a solid, which is a light orange foam. This solid was re-purified by flash chromatography on silica, elution gradient from 0 to 30% methanol in ethyl acetate. The product fractions were concentrated under reduced pressure. The resulting residue was taken up in acetonitrile, filtered, and concentrated under reduced pressure to give 2-fluoro-no. (1- (3fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2, 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4.3- £] isoquinolin-6-yl) pyridin-3-amine (1.03 g, 46%) as a solid, which is a light yellow foam.

¹ H NMR (600 MHz, DMSO-d6, 27 ° C) 1.10 (3H, d), 1.64 (2H, dtt), 2.48 (2H, t), 2.83 (1H, dd) , 2.86 (2H, br d), 2.97 (1H, br dq), 3.02 (1H, dd), 3.45 (1H, dqd), 3.54 (1H, dq), 3, 63 (2H, br s), 3.96 (1H, dquin), 4.44 (2H, dt), 4.91 (1H, s), 6.13 (1H, br d), 6.87 (1H , d), 6.92-7.01 (2H, m), 7.25 (1H, d), 8.06 (1H, s), 13.00 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 495.

Procedures used to prepare (6S, 8R) -6- (5-bromo-6-fluoropyridin-2-yl) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro The -3H-pyrazolo [4,3-f] isoquinoline starting material is described below.

Preparation of tert-butyl (R) - (1- (1H-indazol-4-yl) propan-2-yl) carbamate

- 121 037533

n-Butyllithium in hexane (2.5 M; 96 ml, 241 mmol) was added to a solution of 4-bromo-1H-indazole (24.8 g, 126 mmol) in THF (200 ml) at -78 ° C over 20 min and the mixture was stirred at -78 ° C for 7 hours. Tert-butyl (R) -4-methyl-1,2,3-oxathiazolidine-3-carboxylate-2,2-dioxide (26 g, 110 mmol) was added and the resulting mixture was stirred at -78 ° C for 15 minutes. The cooling bath was removed and the mixture was stirred under these conditions for 18 hours. Aqueous citric acid solution (1N; 130 ml) was added and stirring was continued for 30 minutes. The mixture was extracted with hexanes, and the organic layer was washed with saturated aqueous sodium carbonate solution, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica eluting with 0-60% EtOAc in hexanes to give tert-butyl (R) - (1- (1H-indazol-4-yl) propan-2-yl) carbamate ( 17.3 g, 57%) as a white solid.

Ή NMR (300 MHz, DMSO-d6, 27 ° С) 1.02 (3H, br d), 1.34 (9H, s), 2.82 (1H, br dd), 3.09 (1H, br dd), 3.82 (1H, dt), 6.82 (1H, br d), 6.88 (1H, d), 7.24 (1H, dd), 7.35 (1H, br d), 8.18 (1H, s); 12.97 (1H, s);

mass / charge: ES + [M + H] + = 276.

Preparation of (R) -1- (1H-indazol-4-yl) propan-2-amine dihydrochloride

HCl dioxane (4 M, 100 ml, 400 mmol) was added to a suspension of tert-butyl (R) - (1- (1H-indazol-4yl) propan-2-yl) carbamate (17.3 g, 62.7 mmol) in DCM (200 ml) at room temperature for 10 min. The resulting slurry was stirred overnight and then concentrated under reduced pressure to give (R) -1- (1H-indazol-4-yl) propan-2-amine (15.9 g, 102%) as a white solid.

¹ H NMR (300 MHz, DMSO-d6, 27 ° C) 1.14 (3H, d), 2.97 (1H, dd), 3.40 (1H, dd), 3.45-3.62 ( 1H, m), 6.96 (1H, d), 7.29 (1H, dd), 7.45 (1H, d), 7.97-8.27 (3H, br s), 8.29 ( 1H, d);

mass / charge: ES ⁺ [M + H] ⁺ = 176.

Preparation of (R) -1- (1 H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine

Potassium carbonate (4.75 g, 34.3 mmol) was added to a stirred suspension of (R) -1- (1Hindazol-4-yl) propan-2-amine dihydrochloride salt (2.13 g, 8.58 mmol ), in acetonitrile (25 ml), then 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.191 g, 9.44 mmol) in DCM (12.6 ml) was added dropwise. The mixture was stirred at room temperature for 1.5 days. An additional amount of 2,2,2-trifluoroethyl trifluoromethanesulfonate (398 mg) in DCM (0.3 ml) was then added. The reaction mixture was heated to 60 ° C and after 3 h the reaction mixture was cooled to room temperature and an additional portion of 2,2,2-trifluoroethyl trifluoromethanesulfonate (398 mg) was added as a solution in DCM (0.3 ml). After 18 hours, the reaction mixture was concentrated under reduced pressure to reduce volume and then diluted with DCM. The mixture was 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 chromatography on silica, elution gradient from 0 to 60% ethyl acetate in hexanes, to give (R) -1- (1H-indazol-4-yl) -N- (2,2,2trifluoroethyl) propane -2-amine (2.06 g, 93%) as gum.

Mass / charge: ES ⁺ [M + H] ⁺ = 257.

Preparation of (6S, 8R) -6- (5-bromo-6-fluoropyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

Bg

Trifluoroacetic acid (1.7 ml) was added to a solution of (R) -1- (1H-indazol-4-yl) -N- (2,2,2trifluoroethyl) propan-2-amine (1.83 g, 7.11 mmol) and 5-bromo-6-fluoropicolinaldehyde (1.45 g, 7.11 mmol) in toluene (33.8 ml). The reaction mixture was heated at 90 ° C for 24 h and then concentrated to

- 122 037533 decrease in volume. The mixture was then diluted with dichloromethane and made basic with saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica, elution gradient from 0 to 50% ethyl acetate in hexanes, to give (6S, 8R) -6- (5-bromo-6-fluoropyridin-2-yl) -8-methyl- 7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro3H-pyrazolo [4,3-f] isoquinoline (2.01 g, 64%) as a white solid.

¹ H NMR (300 MHz, DMSO-d6, 27 ° С) 1.11 (3H, d), 2.89 (1H, br dd), 2.94-3.09 (2H, m), 3.31 -3.42 (1H, m), 3.52-3.71 (1H, m), 5.07 (1H, s), 6.96 (1H, d), 7.24-7.36 (2H , m), 8.06 (1H, d), 8.23 (1H, dd), 13.02 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 443.

Example 30.

5-Fluoro-N- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro -3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine

1- (3-Fluoropropyl) azetidin-3-amine (435 mg, 3.29 mmol), (6S, 8R) -6- (5-bromo-3-fluoropyridin-2-yl) -8methyl-7- (2 , 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (730 mg, 1.65 mmol) and sodium tert-butoxide (950 mg, 9.88 mmol) was suspended in 1,4-dioxane (18.3 ml). The mixture was degassed and Brettphos 3G pre-catalyst (149 mg, 0.16 mmol) was added. The reaction mixture was heated to 80 ° C for 1 h. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (10 ml) and washed with water (10 ml). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative LCMS (Waters XSelect CSH C18 column, 5 µm silica, 50 mm in diameter, 100 mm in length) using mixtures of water (containing 1% NH ₃ ) and MeCN with decreasing polarity as eluents. The sample was dissolved in MeOH and separated by SFC using the following chromatographic conditions: column: Phenomonex Lux C1, 30x250 mm, 5 μm, mobile phase: 30% MeOH + 0.1% NH ₃ /70% scCO ₂ to give 5-fluoro -N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro -3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine (396 mg, 49%) as a yellow solid.

Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 1.15 (3H, d), 1.69-1.81 (2H, m), 2.60 (2H, t), 2.77 (1H, dd), 2.92-3.03 (3H, m), 3.14-3.31 (2H, m), 3.65-3.79 (3H, m), 4.04 (1H, q) , 4.43 (2H, t), 4.53 (1H, t), 5.35 (1H, s), 6.54 (1H, dd), 6.76 (1H, d), 7.00 ( 1H, d), 7.66 (1H, d), 7.93 (1H, d), 11.06 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 495.

(6S, 8R) -6- (5-Bromo-3-fluoropyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [ 4,3-f] isoquinoline used as a starting material was synthesized as follows.

(6S, 8R) -6- (5-Bromo-3-fluoropyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [ 4,3-f] isoquinoline

Bg

F— y = N hn-A // ' ^N ~ X nVvJ, \ ^f ·> F ^F

Trifluoroacetic acid (2.13 ml) was added to a solution of (R) -1- (1H-indazol-4-yl) -N- (2,2,2trifluoroethyl) propan-2-amine (1.15 g, 4.47 mmol) and 5-bromo-3-fluoropicolinaldehyde (912 mg, 4.47 mmol) in toluene (42.6 ml) and the resulting mixture was stirred at 100 ° C for 30 min. The reaction mixture was evaporated and the residue was partitioned between DCM (20 ml) and 2M NaOH (20 ml). The layers were separated and the organic phase was concentrated under reduced pressure. The crude product was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give (6S, 8R) -6- (5-6pom-3fluoropyridin-2-yl) -8-methyl-7- (2,2,2-trifluoroethyl) -6.7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (1.15 g, 58%) as a yellow solid.

Ή NMR (500 MHz, CDCl ₃ , 27 ° С) 1.16 (3H, d), 2.86 (1H, dd), 3.01 (1H, dq), 3.19-3.35 (2H, m), 3.68-3.78 (1H, m), 5.42 (1H, s), 6.80 (1H, d), 7.20 (1H, d), 7.59 (1H, dd ), 8.05 (1H, d), 8.27-8.5 (1H, m);

mass / charge: ES ⁺ [M + H] ⁺ 443.

Example 31.

- 123 037533

2,2-Difluoro-3- (6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl3 , 6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol

Tetrabutylammonium fluoride (1.0 M in THF, 0.65 ml, 0.65 mmol) was added to a solution of 6 - ((6S, 8R) -7 (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl ) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinolin-6-yl) -5-fluoro-N- (1- (3-fluoropropyl) azetidin-3-yl ) pyridin-3-amine (320 mg, 0.43 mmol) in THF (2.66 ml) at room temperature and stirred for 64 h. The reaction mixture was evaporated and then dissolved in DMSO and the crude product was purified by flash chromatography with reversed phase (Puriflash, 220 g, C18, 30 µm column) using mixtures of water (containing 1% NH ₃ ) and MeCN with decreasing polarity as eluents. The fractions containing the desired compound were evaporated to dryness to give the crude product (143 mg). The crude product was purified by flash chromatography on silica, elution gradient 0 to 20% MeOH in EtOAc. The fractions containing the product were evaporated to dryness to give 2,2-difluoro-3 - ((6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol (118 mg, 54%) in the form of a mixture of isomers. The material was re-purified by preparative HPLC (Waters XSelect CSH C18 column, 5 μm silica, 30 mm in diameter, 100 mm in length) using mixtures of water (containing 0.1% formic acid) and MeCN with decreasing polarity as eluents. Fractions containing the desired compound were combined and purified by ion exchange chromatography using an SCX-2 column. The desired product was eluted from the column using 1 M NH ₃ / MeOH and the fractions containing the product were evaporated to dryness to give 2,2-difluoro3 - ((6S, 8R) -6- (3-fluoro-5 - ((1 - (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3,6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propane -1-ol (62.1 mg, 29%) as a colorless solid.

¹ H NMR (500 MHz, DMSO, 27 ° C) 1.03 (3H, dd), 1.64 (2H, dq), 2.44 (2H, t), 2.60-2.66 (1H, m), 2.79 (3H, dd), 2.97 (1H, dd), 3.04-3.15 (1H, m), 3.50-3.73 (5H, m), 3.89 -3.97 (1H, m), 4.39 (1H, t), 4.48 (1H, t), 5.21 (1H, s), 5.26 (1H, t), 6.58 ( 1H, d), 6.65-6.71 (2H, m), 7.19 (1H, d), 7.55 (1H, dd), 8.03 (1H, s), 12.94 (1H , s);

mass / charge: ES ⁺ [M + H] ⁺ = 507.

6 - ((6S, 8R) -7- (3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3- f] isoquinolin-6-yl) -5-fluoro-N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine as starting material was synthesized as follows.

(R) -N- (1- (1 H-Indazol-4-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1 amine

3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyl trifluoromethanesulfonate (2.59 g, 5.36 mmol) was added to a solution of (R) -1- (1H-indazol-4-yl) propan-2-amine (0.94 g, 5.36 mmol) and DIPEA (1.39 ml, 8.05 mmol) in 1,4-dioxane (38.9 ml) and the reaction mixture was stirred at 50 ° C for 18 h. the mixture was evaporated, then the residue was diluted with EtOAc and washed with water and the aqueous layer was further extracted with EtOAc. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and evaporated to dryness. The crude product was purified by flash chromatography on silica, elution gradient 0 to 50% EtOAc in heptane. Pure fractions were evaporated to dryness to give (R) -N- (1- (1H-indazol-4-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2difluoropropane-1- amine (1.515 g, 52%) as a colorless gum.

1H NMR (500 MHz, DMSO, 27 ° C) 0.92 (3H, d), 0.97 (9H, s), 1.78-1.86 (1H, m), 2.73 (1H, dd ), 2.98-3.14 (4H, m), 3.83 (2H, td), 6.85 (1H, d), 7.20 (1H, dd), 7.34 (1H, d) , 7.41-7.50 (6H, m), 7.58-7.64 (4H, m), 8.08 (1H, s), 12.98 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 508.

(6R, 8R) -6- (5-Bromo-3-fluoropyridin-2-yl) -7- (3 - ((- butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8methyl-6,7,8, 9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline

- 124 037533

Trifluoroacetic acid (319 μL) was added to a solution of (R) -N- (1- (1H-indazol-4-yl) propan-2yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropane-1 -amine (681 mg, 1.34 mmol) and 5-bromo-3fluoropicolinaldehyde (287 mg, 1.41 mmol) in toluene (6.39 ml) and the resulting mixture was stirred at 110 ° C for 1 h. mixture to room temperature, it was evaporated to dryness and dissolved in DMSO. The crude product was purified by reverse phase flash chromatography (100g Redisep Rf C18 column) using mixtures of water (containing 0.1% formic acid) and MeCN (60-100%) with decreasing polarity as eluents. Fractions containing the desired compound were combined and isolated by ion exchange chromatography using an SCX-2 column. The desired product was eluted from the column using 1 M NH ₃ / MeOH and the pure fractions were evaporated to dryness to give (6S, 8R) -6- (5-bromo-3-fluoropyridin-2-yl) -7- (3- ( (tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (763 mg, 82%) as off-white solid matter.

1H NMR (500 MHz, DMSO, 27 ° C) 0.99 (9H, s), 1.04 (3H, d), 2.74-2.89 (2H, m), 2.97-3.04 (1H, m), 3.31 (1H, s), 3.54-3.62 (1H, m), 3.78 (1H, q), 3.92-4.02 (1H, m), 5.36 (1H, s), 6.72 (1H, d), 7.23 (1H, d), 7.41-7.49 (6H, m), 7.57-7.61 (4H, m), 8.04 (1H, dd), 8.09 (1H, s), 8.38 (1H, d), 13.01 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 693.

6 - ((6S, 8R)) - 7- (3 - ((tert-Butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8-methyl-6,7,8,9-tetrahydro3H-pyrazolo [4.3 -f] isoquinolin-6-yl) -5-fluoro-N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

1- (3-Fluoropropyl) azetidin-3-amine (114 mg,

0.86 mmol), (6S, 8R) -6- (5-bromo-3-fluoropyridin-2-yl) -7 (3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl) -8- methyl 6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline (300 mg, 0.43 mmol) and sodium tert-butoxide (249 mg, 2.59 mmol) were suspended in degassed 1,4 β-dioxane (4.81 ml). Brettphos 3G pre-catalyst (39.2 mg, 0.04 mmol) was added and the mixture was evacuated and purged with nitrogen (x2) and then the reaction mixture was heated to 80 ° C for 45 min. Allow the reaction mixture to cool to room temperature, dilute it with EtOAc and wash with water. The aqueous layer was further extracted with EtOAc and the combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and evaporated to dryness, and used directly without further purification.

Mass / charge: ES + [M + H] ⁺ = 745.

Example 32.

6 - ((6S, 8R) -1-Fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,9,8-tetrahydro-3H-pyrazolo [4,3-f] isoquine -6yl) -N- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

Sodium tert-butoxide (116 mg, 1.20 mmol) and BrettPhos G3 (9.09 mg, 10.04 μmol) were added to a degassed solution of (6S, 8R) -6- (5-bromopyridin-2-yl) -1 -fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3- N-isoquinoline (89 mg, 0.20 mmol) and 1- ( 3-fluoropropyl) azetidin-3-amine (39.8 mg, 0.30 mmol) in 1,4-dioxane (1.00 ml) and the reaction mixture was heated to 90 ° C and stirred for 6 h. After cooling, the reaction the mixture was diluted with EtOAc and water and the layers were separated. The aqueous layer was extracted with EtOAc, then the combined organics were dried over sodium sulfate and evaporated. Purification was carried out by HPLC (Waters CSH C18 OBD column, 5 μm silica, 30 mm in diameter, 100 mm in length) using mixtures of water (containing 0.1% NH ₃ and MeCN as eluents) with decreasing polarity to give the product (42.0 mg, 42%) as a film.

- 125 037533

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.13 (3H, d), 1.69-1.84 (2H, m), 2.61 (2H, td), 2.76 (1H, dd), 2.94 (3H, ddd), 3.16-3.28 (2H, m), 3.45-3.54 (1H, m), 3.67-3.76 (2H, m) , 4.09 (1H, dt), 4.24 (1H, d), 4.43 (1H, td), 4.53 (1H, td), 5.02 (1H, s), 6.76 ( 1H, dd), 6.81 (1H, d), 6.89 (1H, dd), 7.39 (1H, d), 7.81 (1H, d), 11.10 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 495.

(6S, 8R) -6- (5-Bromopyridin-2-yl) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [ 4,3-f] isoquinoline used as a starting material was synthesized as follows.

4-Bromo-3-fluoro-1H-indazole

Selectfluor (49.40 g, 139.6 mmol) was added to a solution of 4-bromo-1H-indazole (25.0 g, 127 mmol) in DMF (254 ml) and the reaction mixture was heated to 70 ° C overnight. After cooling, the reaction mixture was poured into water. The precipitated solid was filtered and dried, then the crude product was purified by flash chromatography on silica, elution gradient from 0 to 30% EtOAc in heptane. Fractions containing product were evaporated to dryness to give 4-bromo-3-fluoro-1H-indazole (4.20 g, 15%) as a pale yellow solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 7.24-7.28 (1H, m), 7.32-7.36 (2H, m), 9.20 (1H, s);

mass / charge: ES ^- [M-H] ^- = 213.

tert-Butyl (R) - (1- (3-fluoro-1H-indazol-4-yl) propan-2-yl) carbamate

n-Butyllithium (1.6 M, 51.9 ml, 83.01 mmol) was added to 4-bromo-3-fluoro-1H-indazole (8.50 g, 39.5 mmol) in THF (124 ml) at -78 ° C and the reaction mixture was stirred for 15 min, heated to -50 ° C for 15 min, then cooled again to -78 ° C. Added tert-butyl (R) -4-methyl-1,2,3oxathiazolidine-3-carboxylate-2,2-dioxide (10.32 g, 43.48 mmol) in THF (20 ml) and the reaction mixture was stirred for 15 min before allowing it to warm to -10 ° C for 30 min. Added 1N. citric acid (200 ml) and the mixture was stirred for 15 min before extraction with EtOAc (x2). The combined organics were dried over magnesium sulfate and evaporated. The crude product was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in heptane. The fractions containing the product were evaporated to dryness to give tert-butyl (R) - (1- (3-fluoro-1H-indazol-4-yl) propan-2-yl) carbamate (5.93 g, 51%) as a colorless solid.

Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 1.17 (3H, d), 1.35 (9H, s), 3.09 (2H, d), 4.00-4.09 (1H, m), 4.41-4.49 (1H, m), 6.97 (1H, d), 7.22 (1H, dd), 7.33 (1H, dd), 9.36 (1H, s );

mass / charge: ES ^- [MH] ^- = 292.

(R) -1- (3-Fluoro-1H-indazol-4-yl) propan-2-amine

4n. hydrochloric acid in dioxane (23.86 ml, 95.45 mmol) was added to tert-butyl (R) - (1 (3-fluoro-1H-indazol-4-yl) propan-2-yl) carbamate (5.60 g, 19.1 mmol) in MeOH (23.9 ml) and the reaction mixture was stirred at room temperature for 2 h. The crude mixture was concentrated, then suspended in EtOAc (100 ml) and washed with saturated aqueous sodium bicarbonate solution (50 ml). The aqueous layer was extracted with EtOAc (x5), then the combined organics were dried over sodium sulfate and evaporated to give (R) -1- (3-fluoro-1Hindazol-4-yl) propan-2-amine (3.10 g, 84%) as a yellow oil that solidified on standing.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.19 (3H, d), 2.86 (1H, dd), 3.09 (1H, dd), 3.33 (1H, dddd), 6.96 (1H, d), 7.22 (1H, dd), 7.33 (1H, dd), 9.93 (1H, s);

mass / charge: ES + [M + H] ⁺ = 194.

(R) -1- (3-Fluoro-1H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine

0.1 M solution of 2,2,2-trifluoroethyl trifluoromethanesulfonate in DCM (25.9 ml, 2.59 mmol) was added to (R) -1- (3-fluoro-1H-indazol-4-yl) propan-2- amine (0.4 g, 2.07 mmol) and DIPEA (0.541 ml, 3.11 mmol) in 1,4-dioxane (20 ml) and the resulting mixture was stirred at 75 ° C overnight. The reaction mixture was concentrated in vacuo and partitioned between ethyl acetate (25 ml) and saturated aqueous solution

- 126 037533 sodium bicarbonate (25 ml). The layers were separated and the aqueous layer was extracted with ethyl acetate (25 ml). The combined organics were washed with saturated aqueous sodium chloride solution (25 ml), dried over sodium sulfate, filtered and concentrated in vacuo. The resulting resin was taken up in methanol and applied to a pre-wetted (methanol) SCX-2 cartridge. The cartridge was washed with methanol and eluted with 1 M ammonia in methanol. The eluate was concentrated in vacuo to give (R) -1- (3-fluoro-1H-indazol-4-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (0.455 g, 80%) in the form of a brown resin.

¹ H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.11 (3H, d), 2.92 (1H, dd), 3.07-3.22 (4H, m), 6.96 (1H , d), 7.25 (1H, dd), 7.34 (1H, dd), 9.50 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 276.

(6S, 8R) -6- (5-Bromopyridin-2-yl) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [ 4,3-f] isoquinoline

Br

F

Trifluoroacetic acid (87 μL) was added to a solution of (R) -1- (3-fluoro-1H-indazol-4-yl) -N- (2,2,2trifluoroethyl) propan-2-amine (100 mg, 0.36 mmol) and 5-bromopicolinaldehyde (67.6 mg, 0.36 mmol) in toluene (0.87 ml) and the resulting mixture was stirred at 90 ° C for 1 h. The reaction mixture was cooled to room temperature and partitioned between DCM ( 5 ml) and saturated aqueous sodium bicarbonate solution (5 ml). The layers were separated and the organic layer was concentrated in vacuo. Purification was carried out by column chromatography on silica gel eluting with 0-50% ethyl acetate in heptane to give (6S, 8R) -6- (5-bromopyridin-2-yl) -1-fluoro-8-methyl-7 (2, 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (89 mg, 55%) as a white solid.

¹ H NMR (500 MHz, CDCl3, 27 ° C) 1.16 (3H, d), 2.92-3.00 (2H, m), 3.253.36 (2H, m), 3.46-3, 52 (1H, m), 5.07 (1H, s), 6.98 (1H, d), 7.09 (1H, dd), 7.43 (1H, d), 7.77 (1H, dd ), 8.55 (1H, dd), 9.09 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 443.

Example 33 5-Fluoro-6 - ((6S, 8R) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3Hpyrazolo [4.3 -f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine

Sodium F tert-butoxide (3.12 g, 32.52 mmol) and BrettPhos G3 (0.245 g, 0.27 mmol) were added to a degassed solution of (6S, 8R) -6- (5-bromo-3-fluoropyridine-2 -yl) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (2.50 g, 5.42 mmol) and 1- (3-fluoropropyl) azetidine-3amine (1.075 g, 8.13 mmol) in 1,4-dioxane (27.1 ml) and the reaction mixture was heated to 60 ° C and stirred for 2 h. After cooling, the reaction mixture was diluted with EtOAc and water and the layers were separated. The aqueous layer was extracted with EtOAc, then the combined organics were dried over sodium sulfate and evaporated. Purification was carried out first by column chromatography on silica gel eluting with 0-100% (10% methanol in ethyl acetate) in heptane, then using a reverse phase Interchim column (0.1% NH ₃ and MeCN as eluents) to afford the product as a mixture of isomers. The products were separated by SFC; the sample was dissolved in MeOH and separated using the following SFC conditions: column: Phenomonex C4, 30x250 mm, 5 μm, mobile phase: 25% MeOH + 0.1% NH ₃ , flow rate: 100 ml / min., BPR 120 bar, column temperature: 40 ° C, to give 5-fluoro-6 - ((6S, 8R) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) 6,7,8,9-tetrahydro -3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridine-3amine (1.780 g, 64%) as a foam.

¹ H NMR (500 MHz, CDCl3, 27 ° C) 1.15 (3H, d), 1.67-1.81 (2H, m), 2.59 (2H, t), 2.81-3, 03 (4H, m), 3.173.34 (2H, m), 3.70 (3H, q), 4.01-4.08 (1H, m), 4.20 (1H, d), 4.43 (1H, t), 4.53 (1H, t), 5.32 (1H, s), 6.54 (1H, dd), 6.81 (1H, d), 6.95 (1H, d) , 7.64-7.68 (1H, m), 9.46 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 513.

(6S, 8R) -6- (5-Bromo-3-fluoropyridin-2-yl) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9- tetrahydro3H-pyrazolo [4,3-f] isoquinoline used as a starting material was synthesized as follows.

(6S, 8R) -6- (5-Bromo-3-fluoropyridin-2-yl) -1-fluoro-8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9- tetrahydro- 127 037533

3H-pyrαzolo [4,3-f] isoquinoline

Br

F

Trifluoroacetic acid (2.16 ml) was added to a solution of (R) -1- (3-fluoro-1H-indαzol-4-yl) -N- (2,2,2trifluoroethyl) propan-2-amine (2.50 g , 9.08 mmol) and 5-bromo-3-fluoropicolinaldehyde (1.85 g, 9.08 mmol) in toluene (43.3 ml) and the resulting mixture was stirred at 90 ° C for 90 min. Allow the reaction mixture to cool and partition it between DCM (50 ml) and saturated aqueous sodium bicarbonate solution (50 ml). The layers were separated and the organic layer was concentrated in vacuo. Purification was carried out by column chromatography on silica gel eluting with 0-50% ethyl acetate in heptane to give (6S, 8R) -6- (5-bromo-3-fluoropyridin-2-yl) -1-fluoro-8-methyl- 7- (2,2,2trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline (2.90 g, 69%) as a white solid.

1H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.15 (3H, d), 2.88-3.04 (2H, m), 3.24-3.35 (2H, m), 3, 63-3.71 (1H, m), 5.39 (1H, s), 6.83 (1H, d), 7.08 (1H, dd), 7.60 (1H, dd), 8.36 (1H, dd), 9.13 (1H, s);

mass / charge: ES ⁺ [M + H] ⁺ = 461.

Examples 34-73 (table below) were prepared using synthetic methods analogous to those described above.

No. example Structure Name 1NYMR LCMS [M + H] 34 ό T F ηχν Λ <(1- (3Fluoropropyl) azetidin-3-yl) -Hmethyl-6 ((65.87?) - 8methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [4, 31] isoquinoline-6yl) pyr idin-3 amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.15 (3H, d), 1.65 - 1.86 (2H, br), 2.59 (2H, t), 2.74 - 2, 84 (1H, br), 2.85 (3H, s), 2.89 3.09 (3H, br), 3.163.41 (2H, br), 3.59 (1H, td), 3.64 - 3.80 (2H, br), 4.10 (1H, q), 4.44 (1H, t), 4.53 (1H, t), 5.06 (1H, s), 6.89 (1H , d), 6.99 (1H, dd), 7.11 (1H, d), 7.30 (1H, d), 7.96-7.98 (2H, m), 10.78 (1H, s). 491 35 ό Yd F jCO% ^F hn γ L <(1- (3Fluoropropyl) azetidin-3-yl) -6 ((6 ^, 87 ^) - 6deutero-8methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [4 , 31] isoquinoline-6yl) pyridin-3 amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.14 (3H, d), 1.76 (2H, dq), 2.62 (2H, t), 2.85 (1H, dd), 2 , 93 - 2.98 (2H, br), 2.99 - 3.04 (1H, br), 3.21-3.31 (2H, br), 3.55 - 3.63 (1H, br) , 3.73 (2H, s), 4.11 (2H, s), 4.44 (1H, t), 4.53 (1H, t), 6.80 (1H, dd), 6.90 ( 1H, d), 7.16 (1H, d), 7.21 (1H, d), 7.84 (1H, d), 8.01 (1H, s) 478 36 HN - ^^ ό Y F WCV HN y * L <(1- (3Fluoropropyl) azetidin-3-yl) -6 ((65.87?) - 1deutero-8methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [4, 31] isoquinoline-6yl) pyr idin-3 amine Ή NMR (500 MHz, CDC1 _3, 27 ° C) 1,14 (ZN, d), 1,71 - 1,81 (2H, br), 2.62 (2H, t), 2,86 (1H, dd), 2.93 3.04 (3H, br), 3.20 3.32 (2H, br), 3.59 (1H, d), 3.73 (2H, s), 4.11 (2H , s), 4.43 (1H, t), 4.53 (1H, t), 5.03 (1H, s), 6.80 (1H, d), 6.90 (1H, d), 7 , 18 (2H, dd), 7.84 (1H, s) 478

- 128 037533

37 HN-Y I ^f £ XO ^{F h} C / 5- (1- (3Fluoropropyl) azetidin-3-yl) -5methoxy-6 ((65,8R) -8methyl-7- (2,2,2trifluoroethyl) - 6,7,8,9tetrahydro-3Npyrazolo [4,3- 1G] isoquinoline-6yl) pyridin-3 amine ^X H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.08 (3H, d), 1.52 - 1.78 (2H, br), 2.46 (2H, t), 2.63 - 2.84 (3H, t), 2.87 - 3.07 (2H, br), 3.34 - 3.50 (1H, br), 3.58 - 3.69 (2H, br), 3 , 80 (3H, s), 3.83 4.07 (2H, br), 4.45 (2H, dt), 5.42 (1H, s), 6.24 (1H, d), 6.54 (1H, d), 6.66 (1H, d), 7.17 (1H, d), 7.26 (1H, d), 8.03 (1H, s), 12.90 (1H, s) ... 507 38 HNό T F zCOC ^ F 5 - ((65.87?) - 1Fluoro-8-methyl-7 (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl ) azet idin-3 yl) pyrazine-2amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.16 (3H, d), 1.68 - 1.85 (2H, br), 2.63 (2H, t), 2.83 (1H , dd), 2.93 2.98 (1H, t), 2.99 3.07 (2H, t), 3.21 (1H, dd), 3.24 - 3.35 (1H, t), 3.49 - 3.54 (1H, t), 3.72 (2H, t), 4.44 (1H, t), 4.47 (1H, d), 4.53 (1H, t), 5 , 05 (1H, s), 5.37 (1H, d), 6.89 (1H, d), 6.96 (1H, dd), 7.75 (1H, d), 8.17 (1H, d) 10.63 (1H, s). 496 39 I z '\ _ ii / = \ -n ~ y // 2-Fluoro-6 ((65.87?) - 1-fluoro8-methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - ( 1- (3fluoropropyl) azet idin-3 yl) pyridin-3 amine Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 1.69 - 1.86 (2H, t), 2.63 (2H, t), 2.83 (1H, dd), 2.91 3.08 (3H, t), 3.133.38 (2H, t), 3.52 (1H, ddd), 3.77 (2H, q), 4.11 (1H, q) , 4.31 4.39 (1H, t), 4.44 (1H, t), 4.53 (1H, t), 4.95 (1H, s), 6.85 (1H, dd), 6 , 89 - 6.98 (2H, t), 7.23 (1H, d), 10.41 (1H, s) 513

- 129 037533

40 HN ^ Su Τ F N = 1 6-Fluoro-L- (1- (3fluoropropyl) azet idin-3-yl) -5 ((65.87 () - 8methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Npyrazolo [ 4.31G] isoquinoline-6yl) pyridine-2amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 1.83 (2H, dp), 2.76 (2H, s), 2.81 2.91 (1H, br), 2.90 3.03 (1H, br), 3.113.32 (4H, br), 3.53 (IH, dt), 3.84 (2H, d), 4.46 (2H, q) , 4.55 (1H, 1), 5.20 (IH, s), 5.98 6.10 (1H, br), 6.83 (IH, d), 6.99 - 7.08 (1H, b), 7.23 (IH, d), 8.06 (IH, d), 10.48 (IH, s). 495 41 I 10 = 4 сО ΠΊ 75- (2-Fluoro-4 ((67 (, 87 () - 8methyl-7- (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Hpyrazolo [4,31G] isoquinoline-6yl) phenyl) -1 - (3fluoropropyl) azet idin-3-amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 1.69 - 1.83 (2H, t), 2.59 (2H, t), 2.77 (IH , dd), 2.84 2.89 (2H, t), 2.89 2.98 (1H, t), 3.03 (IH, dd), 3.21 (1H, dq), 3.38 ( IH, ddd), 3.71 - 3.80 (2H, t), 4.04-4.16 (2H, t), 4.44 (IH, t), 4.53 (1H, t), 4 , 97 (IH, s), 6.41 (IH, t), 6.76 (IH, dd), 6.90 (IH, dd), 6.97 (IH, d), 7.28 (IH, d), 8.06 (IH, d), 10.20 (IH, s). 494 42 τ ϊθ = \ СО # - (1- (3Fluoropropyl) azetidin-3-yl) -5 ((65.87 () - 8methyl-7- (2,2,2trifluoroethyl) - 6,7,8,9- tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) pyrazine-2amine ^X H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.11 (3H, d), 1.49-1.80 (2H, t), 2.42 - 2.48 (2H, t) , 2.76 - 2.93 (3H, t), 2.93 - 3.21 (2H, t), 3.42 - 3.69 (4H, t), 4.23 - 4.34 (1H, t), 4.45 (2H, dt), 4.99 (IH, s), 6.85 (1H, d), 7.25 (IH, d), 7.42 (IH, d), 7, 78 (IH, d), 7.82 (IH, d), 8.06 (1H, s), 12.98 (IH, s). 478

- 130 037533

43 ' ^X N' ^X ^ ό 6 - ((65.87?) - 7- (2,2Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidine- 3-yl) -Nmethylpyridin3-amine Ή NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.06 (3H, d), 1.56 - 1.78 (2H, t), 2.42 - 2.48 (2H, t), 2.55 - 2.70 (1H, t), 2.81 (3H, s), 2.82 - 2.95 (3H, br), 2.96 - 3.23 (2H, t), 3, 39 - 3.54 (1H, t), 3.57 - 3.69 (2H, t), 4.05 (1H, quin), 4.45 (2H, dt), 4.93 (1H, s) , 5.88 (1H, tt), 6.77 (1H, d), 7.07 (2H, d), 7.21 (1H, d), 7.93 (1H, t), 8.05 ( 1H, s), 12.95 (1H, s). 473 44 HN · '^ ό ΧΟΎ X 6 - ((65.8H) -7- (2,2Difluoroethyl) 6,8-dimethyl 6,7,8,9tetrahydro-3Hpyrazolo [4.31] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidine- 3 yl) pyr idin-3 amine Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 1.20 (3H, d), 1.68-1.81 (3H, t), 1.87 (3H, s), 2.59 (2H, t), 2.89 (2H, d), 3.01 (2H, dd), 3.32 (1H, dd), 3.54 - 3.62 (1H, t), 3.68 - 3.75 (2H, t), 3.93 (1H, d), 4.07 (1H, q), 4.43 (1H, t), 4.53 (1H, t), 5.25 5.62 (1H , br), 6.68 (1H, dd), 7.01 (1H, d), 7.08 (1H, d), 7.20 (1H, d), 7.83 (1H, d), 8 , 06 (1H, s), 10.01 (1H, s). 473 45 nyg ^ ό 777 £ CCf hn γ ⁴ X 75- (4 - ((65.85) -7 (2,2Difluoroethyl) 6,8-dimethyl6,7,8,9tetrahydro-3Npyrazolo [4.317] isoquinoline-6yl) -3methoxyphenyl) 1- (3fluoropropyl) azetidin-3 -amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.14 (3H, d), 1.68 - 1.82 (2H, br), 1.85 (3H, s), 2.58 (2H , t), 2.78 2.99 (5H, br), 3.17 (1H, dd), 3.44 (3H, s), 3.49 - 3.60 (1H, br), 3.66 - 3.78 (2H, br), 3.89 (1H, d), 4.03 4.14 (1H, br), 4.48 (2H, dt), 5.26 - 5.54 (1H, br), 5.97 (1H, dd), 6.03 (1H, d), 6.82 502

- 131 037533

(1H, d), 7.00 (1H, d), 7.17 (1H, d), 8.06 (1H, d), 9.95 (1H, s). 46 Τ X 6 - ((65.85) -7- (2,2 Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) -5-fluoro - # - (1 (zfluoropropyl) azet idin-3 yl) pyr idin-3 amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.14 (3H, d), 1.66-1.82 (2H, t), 2.59 (2H, t), 2.72 - 2, 80 (1H, t), 2.85 (1H, dd), 2.90 (2H, d), 2.98-3.12 (1H, t), 3.24 (1H, dd), 3.49 (1H, d), 3.64 - 3.73 (2H, t), 3.98 4.08 (1H, t), 4.16 (1H, d), 4.43 (1H, t), 4 , 53 (1H, t), 5.28 (1H, s), 5.58 - 5.89 (1H, t), 6.52 (1H, dd), 6.82 (1H, d), 7, 19 (1H, d), 7.64-7.69 (1H, t), 8.05 (1H, s), 10.04 (1H, s). 477 47 ΗΝ - ^^ 0 / £ SP 'ην γ 6 - ((65.85) -7- (2,2 Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) -2-fluoro - # - (1 (3fluoropropyl) azet idin-3 yl) pyr idin-3 amine Ή NMR (500 MHz, CDCl ₃ , 27 ° C) 1.12 (3H, d), 1.67-1.81 (2H, t), 2.59 (2H, t), 2.79 (1H, dddd), 2.86 2.93 (2H, br), 3.04 (1H, ddd), 3.33 (1H, dd), 3.53 - 3.62 (1H, br), 3.69 - 3.79 (2H, br), 4.06 (1H, q), 4.21 (1H, d), 4.44 (1H, t), 4.53 (1H, t), 4.89 (1H , s), 5.48 - 5.76 (1H, br), 6.75 (1H, dd), 6.93 (1H, d), 6.98 (1H, d), 7.19 (1H, d), 8.05 (1H, s), 9.99 (1H, s). 477 48 I ^ 4} # - (4 - ((65.85) -7 (2.2- Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31] isoquinoline-6yl) phenyl) -1- (3- Ή NMR (500 MHz, DMSO-d ₆ , 27 ° C) 0.96 (3H, d), 1.52 - 1.64 (2H, br), 2.49-2.61 (1H, br), 2.63 - 2.78 (3H, br), 2.88 - 3.02 (2H, t), 3.26-3.31 (1H, t), 3.53 - 3.64 458

- 132 037533

fluoropropyl) azet idin-3-amine (2H, t), 3.80 - 3.87 (1H, t), 4.38 (2H, dt), 4.74 (1H, s), 5.76 (1H, tt), 5.88 - 5.93 (1H, br), 6.33 (2H, d), 6.70 (1H, d), 6.80 (2H, d), 7.17 (1H, d), 7.98 (1H , s), 12.91 (1H, s). (two peaks corresponding to a hydrogen atom were not observed). 49 HN ' ^X ^ A w A- (4 - ((65.85) -7 (2.2- Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) -3,5d ifluorophenyl) -1 (3fluoropropyl) azet idin-3-amine Ή NMR (500 MHz, DMSO-d ₆ , 27 ° C) 0.95 (3H, d), 1.58 - 1.70 (2H, t), 2.50 - 2.70 (3H, t), 2.79 (1H, br dd), 2.86 - 2.97 (1H, m), 2.95-3.12 (2H, m), 3.09 (1H, br dd), 3.33 - 3.45 (1H, m), 3.71 - 3.88 (2H, m), 3.88-4.01 (1H, m), 4.38 (2H, dt), 5.03 (1H, s), 5.70 (1H, tt), 6.03 (2H, br d), 6.63 (1H, d), 6.69 (1H, br d), 7.13 (1H, d), 7.96 (1H, s), 12.89 (1H, s) 494 fifty A I \ - / \ __ / T 5 - ((65.85) -7- (2,2Difluoroethyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidin-3 yl) pyrazine-2amine ¹ NMR (500 MHz, DMSO-d ₆ , 27 ° C) 1.06 (3H, d), 1.56 - 1.70 (2H, m), 2.42 - 2.47 (2H, m), 2 , 57 - 2.69 (1H, m), 2.75 - 2.87 (3H, m), 2.99-3.13 (2H, m), 3.43-3.51 (1H, m) , 3.54 - 3.62 (2H, m), 4.29 (1H, sxt), 4.43 (2H, dt), 4.94 (1H, s), 5.97 (1H, tt), 6.79 (1H, d), 7.22 (1H, d), 7.41 (1H, d), 7.77 (1H, d), 7.84 (1H, d), 8.04 (1H , s), 12.97 (1H, s) 460

- 133 037533

51 ό JXCf HN y pF 6 - ((65.85) -7- (2,2Difluoroethyl) -8 (difluoromethyl) 6,7,8,9tetrahydro-3Npyrazolo [4,3Cisoquinolin-bil) - # - (1- (3fluoropropyl) azetidin-3 yl) pyridine-3amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.68 1.86 (2H, t), 2.63 (2H, dd), 2.98 (2H, dd), 3.00 - 3, 07 (1H, t), 3.20 (1H, dd), 3.28 - 3.44 (2H, t), 3.70 (3H, q), 4.10 (1H, dh), 4.43 (2H, t), 4.52 (lH, t), 5.19 (1H, s), 5.43 (1H, tdd), 5.81 (1H, td), 6.77 (1H, dd) , 6.84 (1H, d), 7.09 (2H, t), 7.86 (1H, d), 8.00-8.02 (1H, m). 495 52 JT F A- (4 - ((65.8D) -7 ((1- Fluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4,3Cisoquinolin-bil) -3methoxyphenyl) 1- (3fluoropropyl) -Mmethylazetidin3-amine NMR (500 MHz, CDC1 ₃ , 27 ° C) 0.46 0.62 (2H, m), 0.85 1.03 (2H, m), 1.10 (3H, d), 1.66 - 1.89 (2H, m), 2.56-2.62 (2H, m), 2.64 (1H, d), 2.80 (3H, s), 2.89 (1H, dd), 2 , 98 (2H, dt), 3.09 (1H, dd), 3.33 (1H, dd), 3.73 (2H, q), 3.80 (1H, q), 3.87 (3H, s), 4.08 (1H, p), 4.44 (1H, t), 4.53 (1H, t), 5.37 (1H, s), 6.13 (1H, dd), 6, 26 (1H, d), 6.79 (2H, t), 7.07 (1H, d), 8.05 (1H, s), 10.53 (1H, s). 510 53 ko T \ - / \ _ / ABOUT T L '- (2-Fluoro-4 ((65,877) -7 - ((1fluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4,3Cisoquinolin-bil) -5methoxyphenyl) 1- (3fluoropropyl) azet idin-3-amine Lamr (500 MHz, CDC1 ₃ , 27 ° C) 0.46 0.52 (2H, m), 0.92 1.01 (2H, m), 1.08 (3H, d), 1.71 - 1 , 83 (2H, m), 2.55-2.64 (3H, m), 2.84-2.93 (3H, m), 3.02-3.11 (1H, m), 3.35 - 3.42 (1H, m), 3.77 - 3.89 (6H, m), 4.03 - 4.09 (1H, m), 4.12-4.18 (1H, m), 4 , 50 (2H, dt), 514

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5.30 (1H, s), 6.13 (1H, d), 6.65 (1H, d), 6.80 (1H, d), 7.12 (1H, d), 8.05 (1H , s), 9.97 (1H, s). 54 HN - ^^ ό T F HN y * 5 - ((65.87?) - 7 - ((lFluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azeet idin-3 yl) pyridine-2amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 0.47 0.61 (2H, br), 1.04 (2H, d), 1.09 (3H, d), 1.66-1, 81 (2H, br), 2.59 (2H, t), 2.72 (1H, dd), 2.84 (1H, dd), 2.92 (2H, td), 3.01 (1H, dd ), 3.14 (1H, dd), 3.51-3.61 (1H, br), 3.68-3.83 (2H, t), 4.39 (1H, q), 4.43 ( 1H, t), 4.52 (1H, t), 4.92 (1H, d), 4.96 (1H, s), 6.29 (1H, d), 6.88 (1H, d), 7.17 (1H, d), 7.40 (1H, dd), 7.82 (1H, d), 8.04 (1H, d), 11.07 (1H, s). 467 55 I ^L 4J ^> / 'Π 5 - ((65.87?) - 7 - ((1Fluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azet idin-3 yl) pyrazine-2amine ^X H NMR (300 MHz, DMSO-d ₆ , 27 ° C) 0.47 - 0.63 (1H, t), 0.67 0.80 (W, t), 0.81 1.01 (2H, t), 1.04 (3H, d), 1.49 - 1.78 (2H, t), 2.45 (2H, t), 2.57 - 2.94 (4H, t), 2.96 - 3.23 (2H, t), 3.47 - 3.71 (3H, t), 4.22 - 4.34 (1H, t), 4.44 (2H, dt), 4.92 (1H , s), 6.82 (1H, d), 7.22 (Br, d), 7.35 (1H, d), 7.78 (1H, d), 7.82 (1H, d), 8 05 (1H, d), 12.94 (1H, br d). 468

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56 ::. X \ - / \ _ / about MM / 'Ml X Α- (4 - ((6Χ, 87?) - 7 ((1- Fluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) -3methoxyphenyl) 1- (3,3,3trifluoropropyl) azetidine-3-amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 0.48 0.55 (2H, br), 0.90 0.98 (2H, br), 1.09 (3H, d), 2.10- 2.21 (2H, br), 2.57 - 2.72 (3H, br), 2.85 - 2.92 (3H, br), 3.07 (1H, dd), 3.34 (1H, dd), 3.71 - 3.82 (3H, br), 3.86 (3H, s), 4.07 4.14 (1H, br), 5.33 (1H, s), 5.96 ( 1H, dd), 6.11 (1H, d), 6.75 (1H, d), 6.81 (1H, d), 7.11 (1H, d), 8.05 (1H, d), 10.00 (1H, s). 532 57 MM z ' ^z X 6 - ((6 ^, 87 ^) - 7 - ((1Fluorocyclopropyl) methyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31] isoquinoline-6yl) - # - (1- (3fluoropropyl) azet idin-3-yl) -Nmethylpyridin3-amine Ή NMR (300 MHz, DMSO-d ₆ , 27 ° C) 0.40 - 0.57 (1H, br), 0.61 0.77 (1H, br), 0.78 1.00 (2H, br ), 1.02 (3H, d), 1.57 - 1.75 (2H, br), 2.42 - 2.48 (2H, br), 2.66 (1H, dd), 2.80 ( 3N, s), 2.83 - 2.93 (3N, br), 3.00 (1H, dd), 3.19 (1H, br dd), 3.57 - 3.71 (3N, m), 4.04 (1H, quin), 4.45 (2H, dt), 4.88 (1H, s), 6.80 (1H, d), 7.01 -7.10 (2H, m), 7 , 19 (1H, d), 7.93 (1H, d), 8.04 (1H, s), 12.93 (1H, s). 481 58 ώ ην γ ^ν ' 2,2-Difluoro-3 ((6X, 87?) - 6- (5 - ((1 (zfluoropropyl) azet idin-3yl) oxy) pyridin -2-yl) -8-methyl 3,6,8,9 tetrahydro- 7Hpyrazolo [4,3f] ISOCHINOLINE-7- ^X NMR (500 MHz, DMSO, 27 ° C) 1.06 (3H, d), 1.65 (2H, dq), 2.61-2.71 (1H, m), 2.82 (1H, dd ), 2.94 (2H, dd), 3.00 (1H, dd), 3.11 -3.21 (1H, m), 3.36 - 3.42 (1H, m), 3.64 - 3.74 (4H, m), 4.39 (1H, t), 4.48 (1H, t), 4.80 (1H, p), 490

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sludge) propan-1-ol 5.04 (1H, s), 5.39 (1H, t), 6.84 (1H, d), 7.177.26 (3H, t), 8.02 8.07 (2H, t), 12, 97 (1H, S). 59 □: z ' ^z \ __ IL / = 4 GL Ul G' I / Π 2,2-Difluoro-3 ((65.85) -6- (3fluoro-5 - ((1- (3fluoropropyl) azet idin-3 yl) oxy) pyridin -2-yl) -8-methyl 3,6,8 , 9tetrahydro-7Hpyrazolo [4.31G] isoquinoline-7yl) propan-1-ol Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.17 (3H, d), 1.71 - 1.82 (2H, t), 2.65 (2H, t), 2.79 - 2, 95 (3H, t), 3.11 -3.18 (2H, t), 3.19- 3.28 (1H, t), 3.33 (1H, dd), 3.68 (1H, td) , 3.84 (4H, ddt), 4.44 (1H, t), 4.53 (1H, t), 4.79 (1H, p), 5.39 (1H, s), 6.75 ( 1H, d), 6.86 (1H, dd), 7.20 (1H, d), 7.94 (1H, d), 8.06 (1H, d). 508 60 A xxO ^he 2,2-Difluoro-3 ((65.85) -6- (4 - ((1 (zfluoropropyl) azet idin-3 yl) amino) -2methoxyphenyl) 8-methyl-3,6,8,9tetrahydro-7Hpyrazolo [ 4.31G] isoquinoline-7yl) propan-1-ol Ή NMR (500 MHz, CDC1 _3, 27 ° C) 1,13 (ZN, d), 1,39 (1H, h), 1,68 - 1,83 (2H, t), 2.59 (2H, 1), 2.82 2.99 (4H, br), 3.13 (1H, dt), 3.24 (1H, dd), 3.30 - 3.39 (1H, br), 3.56 - 3.86 (8H, br), 4.09 (1H, p), 4.44 (1H, t), 4.53 (1H, t), 5.28 (1H, s), 5.98 (1H , dd), 6.11 (B, d), 6.59 (1H, d), 6.76 (1H, d), 7.16 (1H, d), 8.04 (1H, d). 518 61 ηνΛ ό SHT mM F 2,2-Difluoro-3 ((65.85) -1 -fluoro6- (5 - ((1- (3fluoropropyl) azet idin-3 yl) amino) pyrid in-2-yl) -8methyl-3,6, 8.9 tetrahydro-7Hpyrazolo [4,3f] ISOCHINOLINE-7- ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.15 (3H, d), 1.66-1.88 (2H, br), 2.62 (2H, t), 2.88 (1H , dd), 2.92 3.02 (3H, br), 3.14 (1H, dd), 3.22 (1H, td), 3.37 (1H, ddd), 3.67 - 3.73 (2H, br), 3.76 (1H, td), 3.97 4.15 (2H, br), 4.31 507

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sludge) propan-1-ol (1H, d), 4.44 (1H, t), 4.53 (lH, t), 5.11 (1H, s), 6.72 (1H, dd), 6.79 (1H, d) , 6.87 (1H, d), 7.01 (1H, d), 7.93 (1H, d), 10.19 (1H, s). 62 I o \ 2,2-Difluoro-3 ((65.87?) - 1-fluoro6- (6-fluoro-5 - ((1 (zfluoropropyl) azet idin-3 yl) amino) pyrid in-2-yl) -8methyl- 3,6,8,9tetrahydro-7Hpyrazolo [4.31G] isoquinoline-7yl) propan-1-ol LNMR (500 MHz, CDC1 ₃ , 27 ° C) 1.17 (3H, d), 1.68 - 1.84 (2H, m), 2.62 (2H, t), 2.88 (1H, dd ), 2.92 2.97 (3H, m), 3.08 3.16 (1H, m), 3.163.27 (1H, m), 3.35 3.44 (1H, m), 3.71 3.78 (2H, m), 3.83 (1H, td), 3.93-4.14 (2H, m), 4.38 (1H, d), 4.44 (1H, t), 4 , 53 (1H, t), 5.03 (1H, s), 6.59 6.79 (2H, m), 6.92 (1H, d), 7.09 (1H, dd), 10.06 (1H, s). 525 63 ό HN y 6 - ((65.87?) - 7- (2Fluoro-2methylpropyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidine- 3 yl) pyr idin-3 amine LNMR (500 MHz, DMSO-d ₆ , 27 ° C) 0.94 (3H, d), 1.13 - 1.27 (6H, m), 1.51 - 1.67 (2H, m), 2 , 23 - 2.34 (1H, m), 2.61-2.77 (4H, m), 2.97 (1H, br dd), 3.40 - 3.48 (1H, m), 3, 50 - 3.65 (2H, m), 3.81-3.93 (1H, m), 4.38 (2H, dt), 4.80 (1H, s), 6.11 (1H, br d ), 6.69 - 6.78 (2H, m), 6.95 (1H, d), 7.12 (1H, d), 7.66 (1H, d), 7.96 (1H, s) , 12.86 (1H, s). (two peaks corresponding to a hydrogen atom were not observed). 469

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d), 6.42 (1H, d), 6.65 (1H, d), 7.16 (1H, d), 8.02 (1H, s), 12.93 (1H, s) 69 HN0 „yes 5- (1- (3Fluoropropyl) azetidin-3-yl) -6 ((65.85) -7isobutyl-8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4,3- 1G] isoquinoline-6yl) pyridin-3 amine Ή NMR (300 MHz, DMSO-d ₆ , 27 ° C) 0.73 (ЗН, d), 0.81 (ЗН, d), 0.97 (ЗН, d), 1.51 1.81 (ЗН , t), 1.99 (1H, dd), 2.34 (1H, dd), 2.42 - 2.48 (2H, br), 2.74 (2H, td), 2.85 (1H, dd), 3.09 - 3.22 (1H, br), 3.37-3.54 (1H, t), 3.56-3.71 (2H, t), 3.93 (1H, sxt) , 4.45 (2H, dt), 4.69 (1H, s), 6.12 (1H, d), 6.72 - 6.84 (2H, t), 6.94 (1H, d), 7.16 (1H, d), 7.75 (1H, d), 8.03 (1H, s), 12.90 (1H, s). 451 70 t ν / = \ As? BUT/ 6 - ((65.85) -7- (2,2Difluoropropyl) 8-methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidine- 3 yl) pyr idin-3 amine Ή NMR (300 MHz, DMSO-d ₆ , 27 ° C) 1.07 (3H, d), 1.58 (3H, br t), 1.80-2.05 (2H, m), 2.53 - 2.72 (1H, br), 2.86 (1H, br dd), 2.98 - 3.27 (3H, m), 3.43 3.54 (1H, br), 3.87 (1H , br s), 4.06 (1H, br d), 4.22 - 4.50 (3H, m), 4.52 - 4.65 (2H, m), 4.99 (1H, br s) , 6.56 (1H, br s), 6.80 (1H, d), 6.86 - 7.20 (2H, m), 7.24 (1H, br d), 7.73 - 7.88 (1H, m), 8.08 (1H, s), 9.54 10.04 (1H, m), 12.99 (1H, br s) 473

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64 ο / τ N- (4 - ((6S, 8R) -7 (2-Φτορ-2methylpropyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4,3 ^ isoquinolinyl) -3methoxyphenyl) 1- (3fluoropropyl) azet idin-3-amine ^X H NMR (500 MHz, CDCl ₃ , 27 ° C) 1.05 (3H, d), 1.26 (6H, dd), 1.74 (2H, ddd), 2.37 (1H, dd), 2.54 - 2.64 (2H, br), 2.71-2.9 (4H, br), 3.26 (1H, d), 3.73 (3H, d), 3.84 (3H, s), 3.93 (1H, s), 4.10 (1H, s), 4.43 (1H, t), 4.53 (1H, t), 5.30 (1H, s), 5, 94 (1H, dd), 6.11 (1H, d), 6.67 (1H, d), 6.80 (1H, d), 7.11 (1H, d), 8.04 (1H, d ), 10.06 (1H, S). 498 65 ΗΝ - ^^ ό GOST Isomer Α 6 - ((65.87?) - 7- (2Fluoropropyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4,3P] isoquinoline-6yl) -A- (1- (3fluoropropyl) azetidin-3 yl ) pir idin-3 amine ^X H NMR (500 MHz, DMSO-d ₆ , 27 ° C) 1.00 (3H, d), 1.22 (3H, dd), 1.59 - 1.70 (2H, br), 2.65 - 2.86 (4H, br), 3.14 (1H, br dd), 3.48 (1H, sxt), 3.59 - 3.73 (2H, br m), 3.94 (1H, sxt ), 4.44 (2H, dt), 4.60 (1H, br dsxt), 4.79 (1H, s), 6.19 (1H, br d), 6.74 (1H, d), 6 , 78 (1H, dd), 6.93 (1H, d), 7.16 (1H, d), 7.74 (1H, d), 8.03 (1H, s), 12.93 (1H, s). (The multiplet corresponding to three hydrogen atoms is overlapped by DMSO). 455 66 ΗΝ '' ^ ό Ν = ^ Isomer Β 6 - ((65.87?) - 7- (2Fluoropropyl) -8methyl-6,7,8,9tetrahydro-3Hpyrazolo [4,3P] isoquinoline-6yl) -A- (1- (3fluoropropyl) azetidin-3yl) pir idin-3- ^X H NMR (500 MHz, DMSO-d ₆ , 27 ° C) 1.00 (3H, d), 1.13 (3H, dd), 1.56 - 1.70 (2H, br), 2.28 -2.41 (1H, br), 2.45 (2H, t), 2.72 (2H, q), 2.77 - 2.87 (2H, br), 3.13 (1H, br dd) , 3.46 (1H, sxt), 3.56 3.66 (2H, m), 3.92 455

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71 ζ ΐ) = \ Π 5-Φτορ-Λ- (Ι- (3fluoropropyl) azet idin-3-yl) -6 ((65.85) -8methyl-7- (2,2,3trifluoropropyl) 6,7,8,9tetrahydro-3Npyrazolo [4 , 31Г] isoquinoline-6yl) pyridin-3 amine Ή NMR (500 MHz, DMSO, 27 ° C) 1.04 (3H, d), 1.64 (2H, dq), 2.44 (2H, t), 2.66 2.84 (4H, t) , 2.97 (1H, dd), 3.15-3.26 (1H, t), 3.56 - 3.64 (3H, t), 3.94 (1H, h), 4.39 (1H , t), 4.48 (1H, t), 4.56 - 4.82 (2H, t), 5.19 (1H, s), 6.61 (1H, d), 6.66 - 6, 73 (2H, t), 7.20 (1H, d), 7.55 (1H, dd), 8.04 (1H, s), 12.95 (1H, s). 509 72 ζ _ | ΐ / = \ Π (5) -6- (8.8Dimethyl-7 (2,2,2trifluoroethyl) 6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) - # - (1- (3fluoropropyl) azetidin-3 yl) pir idin-3 amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.06 (3H, s), 1.41 (3H, s), 1.66-1.83 (2H, t), 2.61 (2H, t), 2.93 (3H, dt), 3.13 -3.36 (2H, t), 3.36 - 3.56 (1H, t), 3.71 (2H, q), 4.10 (2H, d), 4.44 (1H, t), 4.53 (1H, t), 4.98 (1H, s), 6.72 (1H, dd), 6.77 (1H, d) , 6.98 (1H, d), 7.11 (1H, d), 7.83 (1H, d), 8.04 (1H, d), 10.72 (1H, s). 491 73 ζ V) = \ ί ' Π (5) -6- (7- (2,2Difluoroethyl) 8,8-dimethyl6,7,8,9tetrahydro-3Hpyrazolo [4.31G] isoquinoline-6yl) -H- (1- (3fluoropropyl) azetidin-3 yl ) pir idin-3 amine Ή NMR (500 MHz, CDC1 ₃ , 27 ° C) 1.04 (3H, s), 1.41 (3H, s), 2.85 - 2.95 (6H, br), 3.21 (2H, t), 3.21 3.35 (2H, br), 3.52 (2H, t), 4.01 (2H, q), 4.44 (1H, t), 4.55 (1H, t) , 4.82 (1H, dt), 4.96 (1H, s), 6.72 (1H, dd), 7.07 (1H, dd), 7.09 7.13 (2H, br), 8 04 (1H, s), 8.20 (1H, s), 10.65 (1H, s). 473

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amine (1H, sxt), 4.44 (2H, dt), 4.55 - 4.72 (1H, br), 4.77 (1H, s), 6.18 (1H, d), 6.73 ( 1H, d), 6.79 (1H, dd), 6.93 (1H, d), 7.16 (1H, d), 7.75 (1H, d), 8.02 (1H, s), 12.92 (1H, s) 67 HN ^^ Isomer A 1- (3- Fluoropropyl) -L '(4 - ((65.87?) - 7- (2fluoropropyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) -3methoxyphenyl) azetidine-3- amine ^X H NMR (400 MHz, DMSO-d ₆ , 27 ° C) 1.01 (3H, d), 1.29 (3H, dd), 1.56 - 1.76 (2H, t), 2.43 - 2.49 (1H, t), 2.57 - 2.66 (1H, t), 2.69 - 2.89 (3H, t), 3.06 (1H, br dd), 3.35 - 3.42 (1H, m), 3.63 3.74 (2H, m), 3.81 (3H, s), 3.89 - 3.99 (1H, m), 4.45 (2H, dt ), 4.70-4.91 (1H, m), 5.17 (1H, s), 5.88 (1H, dd), 6.00 (1H, br d), 6.18 (1H, d ), 6.32 (1H, d), 6.69 (1H, d), 7.19 (1H, d), 8.03 (1H, s), 12.93 (1H, s). (The multiplet corresponding to two hydrogen atoms is overlapped by DMSO). 484 68 LL Z / ^Ш Τ \ - / \ __ / CO / τ 1- (3- Fluoropropyl) -A (4 - ((65.87?) - 7- (2fluoropropyl) -8methyl-6,7,8,9tetrahydro-3Npyrazolo [4.31G] isoquinoline-6yl) -3methoxyphenyl) azetidine-3-amine Ή NMR (400 MHz, DMSO-d ₆ , 27 ° C) 1.00 (3H, d), 1.14 (3H, dd), 1.57 - 1.73 (2H, t), 2.20 - 2.33 (1H, t), 2.42 - 2.48 (2H, t), 2.68 - 2.83 (4H, t), 3.11 (1H, br dd), 3.37 - 3 , 47 (1H, m), 3.59 3.66 (2H, m), 3.81 (3H, s), 3.92 (1H, sxt), 4.45 (2H, dt), 4.63 4.84 (1H, m), 5.14 (1H, s), 5.91 (1H, dd), 5.98 (1H, d), 6.18 (1H, 484

The foregoing description of illustrative embodiments is intended only to familiarize other specialists in the art with the description created by the authors of the present invention, its principles and its practical application, so that others in the art can easily adapt and apply this description in its many forms as they may be best suited to the needs of a particular application. This description and its specific examples illustrating the embodiments of this description are for illustrative purposes only. Therefore, this description is not limited to the illustrative embodiments described herein, and may be modified in various ways. In addition, it should be understood that various features of this specification, which for clarity are described in the context of separate embodiments, can also be combined to form a single embodiment. Conversely, various features of this specification, which for brevity are described in the context of one embodiment, can also be combined to form sub-combinations thereof.

Claims (11)

CLAIM 1. Compound of formula (ID) or a pharmaceutically acceptable salt thereof. where Q represents O or NH; R ¹ is CH2F or CHF2; R ^{6 7 * * * * * * 14 is} selected from the group consisting of: ring Y is selected from the group consisting of: 2. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to claim 1, wherein Q is NH. 3. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to claim 1, wherein Q is O. 4. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to any one of the preceding claims, wherein R ¹ is CH ₂ F. 5. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to any one of the preceding claims, wherein R ^{14 is} selected from the group consisting of: 6. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to any one of the preceding claims, wherein ring Y is selected from the group consisting of: 7. A compound of formula (ID), or a pharmaceutically acceptable salt thereof, according to claim 1, wherein the compound is selected from the group consisting of: N- (4 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine; 6 - ((6S, 8R) -7- (2,2-difluoro-3-methoxypropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine; 6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridine-3-amine; N- (4 - ((6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3- f] isoquinolin-6-yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine; 3 - ((6S, 8R) -6- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -8-methyl-3,6,8, 9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) -2,2-difluoropropan-1-ol; N- (4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6 -yl) -3-methoxyphenyl) -1 - (3-fluoropropyl) azetidine-3-amine; (6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2methoxyphenyl) -8-methyl-6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline; N- (3,5-difluoro-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4, 3-f] -143 037533 isoquinolin-6-yl) phenyl) -1 - (3-fluoropropyl) azetidine-3-amine; 5 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) -4-methoxypyridin-2-amine; N- (4 - ((6S, 8R) -7 - ((3- (fluoromethyl) oxetan-3-yl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Н-pyrazolo [4, 3-f] isoquinolin-6-yl) -3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine; N- (3,5-difluoro-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3Hpyrazolo [4,3-f ] isoquinolin-6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine; (6S, 8R) -7- (2-fluoro-3-methoxy-2-methylpropyl) -6- (4- (1- (3-fluoropropyl) azetidin-3-yloxy) -2methoxyphenyl) -8-methyl-6 , 7,8,9-tetrahydro-3H-pyrazolo [4.3-1 ^: ] isoquinoline; 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8-methyl-3, 6,8,9tetrahydro-7H-pyrazolo [4,3-1 ^: ] isoquinolin-7-yl) propan-1-ol; N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9- tetrahydro3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine; 5-fluoro-6 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline- 6-yl) -N- (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine; N- (3-ethoxy-4 - ((6S, 8R) -7 - ((1-fluorocyclopropyl) methyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3f] isoquinoline -6-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine; N- (4 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline-6- yl) 3-methoxyphenyl) -1- (3-fluoropropyl) azetidin-3-amine; (6S, 8R) -7- (2,2-difluoroethyl) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) -2-methoxyphenyl) -8-methyl6,7, 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline; 3 - ((6S, 8R) -6- (2,6-difluoro-4- (1- (3-fluoropropyl) azetidin-3-ylamino) phenyl) -8-methyl-8,9-dihydro-3Hpyrazolo [4 , 3-f] isoquinoline-7 (6H) -yl) -2-fluoro-2-methylpropan-1-ol; 6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) - N (1- (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine; (6S, 8R) -7- (2,2-difluoroethyl) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) pyridin-2-yl) -8-methyl6,7 , 8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinoline; 1- (3-fluoropropyl) -N- (3-methoxy-4 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7,8,9-tetrahydro- 3Hpyrazolo [4,3-f] isoquinolin-6-yl) phenyl) azetidin-3-amine; 5-fluoro-N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2,2,2-trifluoroethyl) -6,7, 8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine; 2,2-difluoro-3 - ((6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) pyridin-2-yl) -8- methyl3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol; 6 - ((6S, 8R) -7- (2,2-difluoroethyl) -8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) - 5fluoro-N- (1 - (3-fluoropropyl) azetidin-3-yl) pyridin-3-amine; 2,2-difluoro-3 - ((6S, 8R) -6- (5 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) pyridin-2-yl) -8-methyl-3, 6,8,9tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol; 2,2-difluoro-3 - ((6S, 8R) -6- (3-fluoro-5 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) pyridin-2-yl) -8- methyl3,6,8,9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol and 2,2-difluoro-3 - ((6S, 8R) -6- (4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) -2-methoxyphenyl) -8-methyl 3,6,8 , 9-tetrahydro-7H-pyrazolo [4,3-f] isoquinolin-7-yl) propan-1-ol. 8. A compound of formula (ID) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl -7- (2,2,2-trifluoroethyl) -6,7,8,9tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine: or a pharmaceutically acceptable salt thereof. 9. A compound of formula (ID) according to claim 1, wherein the compound is N- (1- (3-fluoropropyl) azetidin-3-yl) -6 - ((6S, 8R) -8-methyl-7- (2 , 2,2-trifluoroethyl) -6,7,8,9-tetrahydro-3H-pyrazolo [4,3-f] isoquinolin-6-yl) pyridin-3-amine: - 144 037533 10. The use of a compound or a pharmaceutically acceptable salt thereof according to claim 8 for the prevention or treatment of cancer in a warm blooded animal. 11. Use according to claim 10, wherein the cancer is breast cancer or gynecological cancer.