EA038835B1 - TETRAHYDRO-PYRIDO[3,4-b]INDOLE ESTROGEN RECEPTOR MODULATORS AND USES THEREOF - Google Patents

Abstract

Described herein are tetrahydro-pyrido[3,4-b]indol-1-yl compounds with estrogen receptor modulation activity or function having the Formula I structureand stereoisomers, tautomers or pharmaceutically acceptable salts thereof, and with the substituents and structural features described herein. Also described are pharmaceutical compositions and medicaments that include the Formula I compounds, as well as methods of using such estrogen receptor modulators, alone and in combination with other therapeutic agents, for treating diseases or conditions that are mediated or dependent upon estrogen receptors.

Description

Scope of invention

This document describes compounds, including pharmaceutically acceptable salts thereof, pharmaceutical compositions containing such compounds, and methods of treating such compounds for diseases or conditions that are sensitive to estrogen, dependent on estrogen receptors or mediated by estrogen receptors, such as breast cancer, lung cancer, ovarian cancer, endometrial cancer, prostate cancer, or uterine cancer.

State of the art

The estrogen receptor (ER) is a ligand-activated transcriptional regulatory protein that mediates the induction of various biological effects through its interaction with endogenous estrogens. Endogenous estrogens include β-estradiol and estrone. ER was found to have two isoforms: ER-α (alpha) and ER-β (beta). Estrogens and estrogen receptors are implicated in a number of diseases or conditions, such as breast cancer, lung cancer, ovarian cancer, colon cancer, prostate cancer, endometrial cancer, uterine cancer, and other diseases or conditions. There is a need for new ER-α targeting agents that have activity against metastatic disease and acquired resistance.

The essence of the invention

This invention relates generally to compounds of tetrahydropyrido [3,4-b] indol-1-yl with modulating activity or function of an estrogen receptor having the structure of formula I

as well as their stereoisomers or pharmaceutically acceptable salts, and with the substituents and structural features described herein.

One aspect of this invention is a pharmaceutical composition having estrogen receptor modulator activity, consisting of a compound of Formula I, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

One aspect of the present invention is a method of treating an ER-associated disease or disorder in a patient, such as breast cancer, lung cancer, ovarian cancer, endometrial cancer, prostate cancer or uterine cancer, comprising administering to the patient a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

In the following, reference will be made in detail to certain embodiments of the invention, examples of which are illustrated in the accompanying structures and claims. While the invention will be described in conjunction with the recited embodiments, it should be understood that the invention is not limited to these embodiments. On the contrary, this invention covers all alternatives, modifications and equivalents that may be included within the scope of this invention as defined by the claims. A person skilled in the art will appreciate numerous methods and materials similar or equivalent to those described herein that can be used in the practice of this invention. This invention is in no way limited to the described methods and materials. In the event that one or more of the included literature, patents and similar materials differs from or contradicts this application, including, but not limited to, certain terms, the use of terms, described techniques or the like, the control is this application. Unless otherwise indicated, all technical and scientific terms used in this document have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. While methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned in this document are incorporated by reference in their entirety. The nomenclature used in this application is based on the IUPAC systematic nomenclature, unless otherwise indicated.

Definitions.

When referring to the number of substituents, the term one or more refers to the range from one substituent to the maximum possible number of substitutions, i. E. from replacing one hydrogen to replacing all hydrogen atoms with substituents. The term “substituent” means an atom or group of atoms that replaces a hydrogen atom on the parent molecule. The term substituted means that the specified group bears one or more substituents. If any group may carry more than one substituent, and

- 1038835 provides a variety of possible substituents, these substituents are independently selected and do not have to be the same. The term unsubstituted means that the specified group bears no substituents. The term optionally substituted means that said group is unsubstituted or substituted with one or more substituents independently selected from the group of possible substituents. When indicating the number of substituents, the term one or more means the range from one substituent to the maximum possible number of substitutions, i. E. from replacing one hydrogen to replacing all hydrogen atoms with substituents.

The term alkyl, as used herein, refers to a saturated linear or branched monovalent hydrocarbon radical of one to twelve carbon atoms (C ₁ -C ₁₂ ), where the alkyl radical may optionally be independently substituted with one or more substituents, described below. In another embodiment, the alkyl radical contains from one to eight carbon atoms (C ₁ -C ₈ ) or from one to six carbon atoms (C ₁ -C ₆ ). Examples of alkyl groups include, but are not limited to, methyl (Me, —CH ₃ ), ethyl (Et, —CH ₂ CH ₃ ), 1-propyl (n-Pr, npropyl, —CH ₂ CH ₂ CH ₃ ), 2 -propyl (i-Pr, i-propyl, -CH (Ch ₃ ) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ Ch ₂ CH ₂ CH ₃ ), 2methyl-1-propyl (i -Bu, i-butyl, -CH ₂ CH (CH ₃ ) ₂ ), 2-butyl (s-Bu, sec-butyl, -CH (CH ₃ ) CH ₂ CH ₃ ), 2-methyl-2propyl (t- Bu, tert-butyl, -C (CH ₃ ) ₃ ), 1-pentyl (n-pentyl, CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH (CH ₃ ) CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH (Ch ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C (CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH ( CH ₃ ) CH (CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH (CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH (CH ₃ ) CH ₂ CH ₃ ), 1hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH (CH3) CH2CH2CH2CH3), 3-hexyl (-CH (CH2CH3) (CH2CH2CH3)), 2-methyl-2-pentyl (-C (CH3) 2CH2CH2CH3 ), 3-methyl-2-pentyl (-CH (CH3) CH (CH3) CH2CH3), 4-methyl-2-pentyl (-CH (CH ₃ ) CH ₂ CH (CH ₃ ) ₂ ), 3-methyl- 3-pentyl (-C (CH ₃ ) (CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH (CH2CH3) CH (CH3) 2), 2,3-dime tyl-2-butyl (-C (CH3) 2CH (CH3) 2), 3,3-dimethyl-2-butyl (-CH (CH3) C (CH ₃ ) ₃ , 1-heptyl, 1-octyl, etc.) P.

The term alkyldiyl, as used herein, refers to a saturated linear or branched bivalent hydrocarbon radical of about one to about twelve carbon atoms (C ₁ -C ₁₂ ) in length, where the alkyldiyl radical may optionally be independently substituted with one or more substituents described below. In another embodiment, the alkyldiyl radical contains from one to eight carbon atoms (C ₁ -C ₈ ) or from one to six carbon atoms (C ₁ -C ₆ ). Examples of alkyldiyl groups include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), and the like. An alkyldiyl group can also be called an alkylene group.

The term alkenyl refers to a linear or branched monovalent hydrocarbon radical with a length of two to eight carbon atoms (C ₂ -C ₈ ) containing at least one unsaturated, i. E. a carbon-carbon double sp ² bond, where an alkenyl radical may optionally be independently substituted with one or more of the substituents described herein and includes radicals having cis and trans orientations, or alternatively E- and Z '' - orientations. Examples include, but are not limited to, ethylene or vinyl (-CH = CH ₂ ), allyl (-CH ₂ CH = CH ₂ ), and the like.

The terms alkenylene or alkenyldiyl refer to a linear or branched divalent hydrocarbon radical of two to eight carbon atoms in length (C ₂ -C ₈ ) containing at least one unsaturated, i. E. a carbon-carbon, sp ² double bond, where the alkenylene radical may optionally be independently substituted with one or more substituents described herein and includes radicals having cis and trans orientations or, alternatively, E - and Z '' - orientations. Examples include, but are not limited to, ethyleneylene or vinylene (-CH = CH-), allyl (-Ch ₂ CH = CH-), and the like.

The term alkynyl refers to a linear or branched monovalent hydrocarbon radical of two to eight carbon atoms in length (C2-C8) containing at least one unsaturated, i. E. a carbon-carbon, sp-triple bond, where an alkynyl radical may optionally be independently substituted with one or more of the substituents described herein. Examples include, but are not limited to, ethynyl ((—C ^ CH), propynyl (propargyl, —CH ₂ C ^ CH), and the like.

The terms alkynylene or alkynyldiyl refer to a linear or branched divalent hydrocarbon radical of two to eight carbon atoms in length (C ₂ -C ₈ ) containing at least one unsaturated, i. E. carbon-carbon, triple sp-bond, where the alkynylene radical may optionally be independently substituted with one or more of the substituents described herein. Examples include, but are not limited to, ethynylene (—C = C—), propynylene (propargylene, —CH ₂ C═C—), and the like.

The terms carbocycle, carbocyclyl, carbocyclic ring and cycloalkyl refer to a monovalent non-aromatic, saturated or partially unsaturated ring containing 3 to 12 carbon atoms (C ₃ -C ₁₂ ) as a monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycles containing from 7 to 12 atoms can be organized, for example, in the form of a bicyclo [4.5] -, [5.5] -, [5.6] - or [6.6] -system, and bicyclic carbocycles,

- 2,038835 containing 9 or 10 ring atoms, can be organized in the form of bicyclo [5,6] - or [6,6] systems or in the form of bridging systems such as bicyclo [2,2,1] heptane, bicyclo [2 , 2.2] octane; and bicyclo [3.2.2] nonane. Spirocarbocyclic moieties are also included within the scope of this definition. Examples of spirocarbocyclic moieties include [2,2] pentanyl, [2,3] hexanyl, and [2,4] heptanyl. Examples of monocyclic carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1- cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl and the like. Carbocyclic groups may optionally be independently substituted with one or more of the substituents described herein.

The term carbocyclyldiyl refers to a divalent non-aromatic, saturated or partially unsaturated ring containing 3 to 12 carbon atoms (C ₃ -C ₁₂ ) as a monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring.

The term aryl means a monovalent aromatic hydrocarbon radical of 620 carbon atoms (C ₆ -C ₂₀ ) obtained by removing one hydrogen atom from one carbon atom of the parent aromatic ring system. Certain aryl groups are represented in illustrative structures as Ar. Aryl includes bicyclic radicals containing an aromatic ring fused to a saturated, partially unsaturated ring or aromatic carbocyclic ring. Typical aryl groups include, but are not limited to, radicals derived from benzene (phenyl), substituted benzenes, naphthalene, anthracene, biphenyl, indenyl, indanyl, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, etc. P. Aryl groups may optionally be independently substituted with one or more of the substituents described herein.

The terms arylene or aryldiyl mean a bivalent aromatic hydrocarbon radical of 6 to 20 carbon atoms (C ₆ -C ₂₀ ) obtained by removing two hydrogen atoms from two carbon atoms of the parent aromatic ring system. Certain aryldiyl groups are represented in illustrative structures as Ar. Aryldiyl includes bicyclic radicals containing an aromatic ring fused to a saturated, partially unsaturated ring or aromatic carbocyclic ring. Typical aryldiyl groups include, but are not limited to, radicals derived from benzene (phenyldiyl), substituted benzenes, naphthalene, anthracene, biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, etc. P. Aryldiyl groups are also referred to as arylenes and may optionally be substituted with one or more of the substituents described herein.

The terms heterocycle, heterocyclyl, and heterocyclic ring are used interchangeably herein and refer to a saturated or partially unsaturated (i.e., containing one or more double and / or triple bonds within the ring) carbocyclic radical containing from 3 to 20 ring atoms, in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and sulfur; the remaining ring atoms are C, in which one or more ring atoms are optionally independently substituted with one or more substituents described below. The heterocycle can be a monocycle containing 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P and S), or a bicycle containing 7 to 10 ring members ( 4 to 9 carbon atoms and 1 to 6 heteroatoms selected from N, O, P and S), for example bicyclo [4.5] -, [5.5] -, [5.6] or [6.6] -system. Heterocycles are described in Paquette Leo A. Principles of Modern Heterocyclic Chemistry (W.A. Benjamin, New York, 1968), in particular Ch. 1, 3, 4, 6, 7 and 9; The Chemistry of Heterocyclic Compounds, A series of Monographs (John Wiley & Sons, New York, 1950 to the present), in particular in Vol. 13, 14, 16, 19 and 28; and J. Am. Chem. Soc. (1960) 82: 5566. The term heterocyclyl also includes radicals where heterocyclic radicals are fused to a saturated, partially unsaturated ring or aromatic carbocyclic or heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, morpholin-4-yl, piperidin-1-yl, piperazinyl, piperazin-4-yl-2-one, piperazin-4-yl-3-one, pyrrolidin-1-yl, thiomorpholin-4-yl, S-dioxothiomorpholin-4-yl, azocan-1-yl, azetidin-1-yl, octahydropyrido [1,2-a] pyrazin-2-yl, [1,4] diazepan-1-yl , pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinil, azetidinyl, thietynyl -pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanil, 1,3-dioxolanyl, pyrazolinyl, dithianil, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl, 3,0abycyloxyl, 3,1 -azabicyclo [4.1.0] heptanyl, azabicyclo [2.2.2] hexanyl, 3H-indolyl-quinolizinyl and n-pyridylureas. Spiroheterocyclic moieties are also included within the scope of this definition. Examples of spiroheterocyclic moieties include azaspiro [2.5] octanyl and azaspiro [2.4] heptanyl. Examples of a heterocyclic group in which two ring atoms are substituted with oxo (= O) moieties are pyrimidinonyl and 1,1dioxothiomorpholinyl. Heterocyclic groups can possibly be independently substituted with one

- 3 038835 or more than one of the substituents described in this document.

The term heterocyclyldiyl refers to a divalent, saturated, or partially unsaturated (i.e., having one or more double and / or triple bonds in the ring) carbocyclic radical of 3-20 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and sulfur, with the remaining ring atoms being C, where one or more of the ring atoms is optionally independently substituted with one or more of the described substituents.

The term heteroaryl refers to a monovalent aromatic radical containing 5-, 6 or 7-membered rings and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms containing one or more heteroatoms independently selected from nitrogen, oxygen and sulfur atoms. Examples of heteroaryl groups are pyridinyl (including, for example, 2-hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4-hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, oxazyl, thiazolyl , isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl , quinazolinyl, quinoxalinyl, naphthyridinyl and furopyridinyl. Heteroaryl groups may optionally be independently substituted with one or more of the substituents described herein.

The term heteroaryldiyl refers to a divalent aromatic radical containing 5-, 6-, or 7-membered rings and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms containing one or more heteroatoms, independently chosen from nitrogen, oxygen and sulfur atoms.

Heterocyclic or heteroaryl groups can be linked through a carbon atom (carbon-linked) or through a nitrogen atom (nitrogen-linked), where possible. By way of example and without limitation, carbon-linked heterocycles or heteroaryls are linked at position 2, 3, 4, 5, or 6 of pyridine, at position 3, 4, 5, or 6 of pyridazine, at position 2, 4, 5, or 6 of pyrimidine, at position 2, 3, 5 or 6 pyrazine, at position 2, 3, 4 or 5 of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, at position 2, 4 or 5 of oxazole, imidazole or thiazole, at position 3, 4 or 5 isoxazole, pyrazole or isothiazole, in position 2 or 3 of aziridine, in position 2, 3 or 4 of azetidine, in position 2, 3, 4, 5, 6, 7 or 8 of quinoline or in position 1, 3, 4, 5, 6 , 7 or 8 isoquinoline.

By way of example and without limitation, nitrogen-linked heterocycles or heteroaryls are linked at position 1 of aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2- pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, III-indazole, at position 2 of isoindole or isoindoline, at position 4 of morpholine and at position 9 of carbazole or β-carboline.

The terms treat and treat refer to therapeutic treatment, the purpose of which is to slow down (reduce) an unwanted physiological change or disorder, such as the development or spread of arthritis or cancer. For the purposes of this invention, significant or desirable clinical results include, but are not limited to, amelioration of symptoms, a decrease in the severity of the disease, a stable (i.e., no worsening) state of the disease, a delay or slowdown in the progression of the disease, an improvement or temporary relief of the disease state, and remission ( both partial and general), both detectable and undetectable. Treatment can also mean an increase in survival over the estimated survival rate without treatment. Individuals in need of treatment include individuals with the condition or disorder.

The phrase therapeutically effective amount means an amount of a compound of this invention that (i) treats a particular disease, condition or disorder, (ii) reduces, alleviates, or eliminates one or more symptoms of a particular disease, condition or disorder, or (iii) prevents or delays the development of one or more symptoms of a particular disease, condition, or disorder described herein. In the case of cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce the size of the tumor; inhibit (ie, to some extent slow down and preferably stop) the infiltration of cancer cells of peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or to some extent relieve one or more of the symptoms associated with cancer. The drug can prevent the growth and / or kill existing cancer cells to some extent, it can be cytostatic and / or cytotoxic. In cancer treatment, efficacy can be determined, for example, by assessing the time to disease progression (TTP) and / or by determining the response rate (RR).

The term cancer refers to or describes a physiological condition in mammals that is typically characterized by uncontrolled cell growth. The tumor contains one or more

- 4 038835 one cancer cell. Examples of cancers include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia (leukemia) or lymphoid cancer. More specific examples of such cancers include squamous cell carcinoma (eg, epithelial squamous cell carcinoma), lung cancer including small cell lung cancer, non-small cell lung cancer (NSCLC), adenocarcinoma of the lung and squamous cell lung carcinoma, peritoneal cancer, hepatocellular carcinoma , stomach or gastrointestinal tract cancer, including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, carcinoma of the uterus or endometrium, carcinoma of the salivary glands, kidney or renal cancer, prostate cancer, vaginal cancer, thyroid cancer, liver carcinoma, carcinoma of the anal canal, carcinoma of the penis, and head and neck cancer.

Hematologic malignancies are types of cancer that affect the blood, bone marrow, and lymph nodes. Because all three of these tissues are closely linked through the immune system, a disease that affects one of these three tissues often affects others as well: although lymphoma is a disease of the lymph nodes, it often spreads to the bone marrow, affecting the blood. Hematologic malignancies are malignant neoplasms (cancers) and are usually treated by specialists in hematology and / or oncology. In some centers, hematology / oncology is one division of internal medicine, while in others they are considered separate divisions (there are also surgeons-oncologists and radiation oncologists). Not all hematologic disorders are malignant (cancerous); these other blood conditions may also be monitored by a hematologist. Hematologic malignancies can arise from either of two main blood cell lines: myeloid and lymphoid cell lines. The myeloid cell line usually produces granulocytes, erythrocytes, platelets, macrophages, and mast cells; the lymphoid cell line produces B-, T-, NK- and plasma cells. Lymphomas, lymphocytic leukemias, and myelomas originate from the lymphoid lineage, while acute and chronic myelogenous leukemias, myelodysplastic syndromes, and myeloproliferative diseases are of myeloid origin. Leukemias include acute lymphoblastic leukemia (ALL, acute lymphoblastic leukemia, ALL), acute myelogenous leukemia (AML, acute myelogenous leukemia, AML), chronic lymphocytic leukemia (CLL, chronic lymphocytic leukemia, myelogenous , LSU), acute monocytic leukemia (AMOL) and small lymphocytic lymphoma (SLL). Lymphomas include Hodgkin's lymphoma (all four subtypes) and non-Hodgkin's lymphoma (NHL, Non-Hodgkin's lymphoma, NHL, all subtypes).

A chemotherapeutic agent is a chemical compound used to treat cancer, regardless of the mechanism of action. Classes of chemotherapeutic agents include, but are not limited to, alkylating agents, antimetabolites, mitotic spindle blocking plant alkaloids, cytotoxic / antitumor antibiotics, topoisomerase inhibitors, antibodies, photosensitizers, and kinase inhibitors. Chemotherapeutic agents include compounds used in targeted therapy and conventional chemotherapy. Examples of chemotherapeutic agents include ibrutinib (IMBRUVICA ™, APCI-32765, Pharmacyclics Inc./Janssen Biotech Inc .; CAS per. No. 936563-96-1, US 7514444), idalisib (ZYDELIG®, CAL-101, GS 1101, GS- 1101, Gilead Sciences Inc .; CAS per. No. 1146702-54-6), erlotinib (TARCEVA®, Genentech / OSI Pharm.), Docetaxel (TAXOTERE®, Sanofi-Aventis), 5-FU (fluorouracil, 5-fluorouracil, CAS per. No. 51-21-8), gemcitabine (GEMZAR®, Lilly), PD-0325901 (CAS No. 391210-10-9, Pfizer), cisplatin (Platinol®, (SP-4-2) diaminedichloroplatin (P) , cis-diamine, dichloroplatinum (P), CAS # 15663-27-1), carboplatin (CAS # 41575-94-4), paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ), trastuzumab ( HERCEPTIN®, Genentech), temozolomide (4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo [4,3,0] -nona 2,7,9-triene-9-carboxamide, CAS No. 85622-93-1, TEMODAR®, TEMODAL®, Schering Plow), tamoxifen ((Z) -2- [4- (1,2-diphenylbut-1-enyl) phenoxy] -N, N-dimethylethanamine, NOLVADEX®, ISTUBAL®, VALODEX®) and doxorubicin (ADRIAMYCIN®, CAS No. 232 14-92-8), Akti-1/2, HPPD and rapamycin.

Chemotherapeutic agents include inhibitors of B-cell receptor targets such as BTK inhibitors Bcl-2 and JAK.

Other examples of chemotherapeutic agents include the following: oxaliplatin (ELOXATIN®, Sanofi), bortezomib (VELCADE®, Millennium Pharm.), Sutent (SUNITINIB®, SU11248, Pfizer), letrozole (FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novart ), XL-518 (Mek inhibitor, Exelixis, WO 2007/044515), ARRY-886 (Mek inhibitor, AZD6244, Array BioPharma, Astra Zeneca), SF-1126 (PI3K inhibitor, Semafore Pharmaceuticals), BEZ-235 (PI3K inhibitor , Novartis), XL-147 (PI3K inhibitor, Exelixis), PTK787 / ZK 222584 (Novartis), fulvestrant (FASLODEX®, AstraZeneca), leucovorin (folic acid), rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib ( , GSK572016, Glaxo Smith Kline), lonafarnib (SARASAR ™, SCH 66336, Schering Plow), sorafenib (NEXAVAR®, BAY43-9006, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), irinotecan (CAMPTOSAR®, CPT-11

- 5 038835

Pfizer), Tipifarnib (ZARNESTRA ™, Johnson & Johnson), ABRAXANE ™ (Cremophor Free), Albumin-Engineered Paclitaxil Nanoparticles (American Pharmaceutical Partners, Schaumberg, II), Vandetanib (rINN, ZD6474, ZACTIMA®, AstraZenecil) , AG1478, AG1571 (SU 5271; Sugen), temzirolimus (TORISEL®, Wyeth), pazopanib (GlaxoSmithKline), canfosfamide (TELCYTA®, Telik), thiotepa and cyclophosphamide (CYTOXAN®, NEOSAR®); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); camptothecin (including the synthetic analogue topotecan); bryostatin; callistatin; CC-1065 (including its synthetic analogs adoselesin, carzelesin and bisellesin); cryptophycins (in particular, cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pankratistatin; sarcodictin; spongystatin; nitrogen mustards such as chlorambucil, chloronaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicin, fenesterin, prednimustine, trophosphamide, uraciliprit; nitrosoureas such as carmustine, chlorzotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as enedyne antibiotics (eg, calicheamicin, calicheamicin gamma II, calicheamicin omega II (Angew Chem. Intl. Ed. Engl. (1994) 33: 183-186); dynemycin, dynemycin A; bisphosphonates such as clodronate; esperamicin ; as well as the chromophore neocarzinostatin and related chromoprotein chromophores of enedyne antibiotics), aclacinomisins, actinomycin, anthramycin, azaserin, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycin-5-dactin-oxycin-5 , morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxidoxorubicin, epirubicin, ezorubicin, idarubicin, nemorubicin, marcellomycin, mitomycins, such as mitomycin C, nogalamycinamyrcinoma, olivycin , streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacytidine, 6-azauridine, karmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostan, testolactone; antiadrenal drugs such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid; aceglatone, aldophosphamide glycoside; aminolevulinic acid; enilluracil; amsacrine; bestrabucil; bisanthrene; edatraxate; defopamine; demecolcine; diaziquon; elfornithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguason; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pyirubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; polysaccharide complex PSK® (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizophyran; spirogermanium; tenuazonic acid; triaziquon; 2,2 ', 2-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidin); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (Ara-C); cyclophosphamide; thiotepa; 6-thioguanine; meracptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP16); ifosfamide; mitoxantrone; vincristine; Vinorelbine (NAVELBINE®); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capetacibin (XELODA®, Roche); ibandronate; CPT-11; RFS 2000 topoisomerase inhibitor; difluoromethylornithine (DFMO); retinoids such as retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above.

Also, the definition of a chemotherapeutic agent includes: (i) antihormonal agents that regulate or inhibit the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX®; tamoxifen citrate ), raloxifene, droloxifene, 4-hydroxy tamoxifen, trioxifene, keoxifen, LY117018, onapristone and FARESTON® (toremifene citrate), and selective estrogen receptor modulators (SERD) such as fulvestrant®, AASLODEX (ii) aromatase inhibitors that inhibit the aromatase enzyme that regulates the production of estrogens in the adrenal glands, such as, for example, 4 (5) -imidazoles, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestan, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole; Novartis) and ARIMIDEX® (anastrozole; AstraZeneca); (iii) antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; as well as troxacitabine (1,3-dioxolane nucleosine cytosine analog); (iv) protein kinase inhibitors such as MEK inhibitors such as cobimetinib (WO 2007/044515); (v) lipid kinase inhibitors such as tazelizib (GDC-0032, Genentech Inc.); (vi) antisense oligonucleotides, in particular those that inhibit the expression of genes in signaling pathways involved in aberrant cell proliferation, for example PKC-α, Ralf and H-Ras, such as oblimersen (GENASENSE®, Genta Inc.); (vii) ribozymes such

6 038835 as inhibitors of VEGF expression (eg ANGIOZYME®) and inhibitors of HER2 expression; (viii) vaccines such as gene therapy vaccines such as ALLOVECTIN®, LEUVECTIN® and

VAXID®; rIL-2 PROLEUKIN®; topoisomerase 1 inhibitors such as LURTOTECAN®; rmRH ABARELIX®; (ix) anti-angiogenic agents such as bevacizumab (AVASTIN®, Genentech); and pharmaceutically acceptable salts, acids and derivatives of any of the above.

Also, the definition of a chemotherapeutic agent includes therapeutic antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech / Biogen Idec), pertuzumab (PERJETA ™, 2C4, Genentech), trastuzumab (HERCEPTIN®, Genentech), trastuzumab emtansinA Inc. (KADC BEXXAR, Corixia).

A metabolite is a product formed during the metabolism of the specified compound or its salt in the body. Compound metabolites can be determined using standard techniques known in the art, and their activities determined using assays such as those described herein. Such products can be formed, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic degradation, and the like. of the introduced compound. Accordingly, this invention includes metabolites of the compounds of this invention, including compounds obtained by a process comprising contacting a compound of Formula I of this invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

The term package insert refers to instructions that are usually included in commercial packages of therapeutic products and contain information about indications, use, dosage, administration, contraindications and / or warnings regarding the use of such therapeutic products.

The term chiral refers to molecules that have the property of non-superimposing mirror images of partners, while the term achiral refers to molecules whose mirror images can be superimposed on images of their partners.

The term stereoisomers refers to compounds that have the same chemical structure, but differ in the arrangement of atoms or groups in space.

The term diastereomers refers to stereoisomers with two or more chiral centers, whose molecules are not mirror images of each other. Diastereomers have various physical properties, such as melting points, boiling points, spectral properties, and reactivity. Mixtures of diastereomers can be separated using high-resolution analytical procedures such as electrophoresis and chromatography.

The term enantiomers refers to two stereoisomers of a compound that are not mirror images that could be superimposed on each other.

The stereochemical definitions and conventions used in this invention generally correspond to S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers and therefore can exist in various stereoisomeric forms. It is understood that all stereoisomeric forms of the compounds of this invention, including but not limited to diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of this invention. Many organic compounds exist in optically active forms, i.e. they have the ability to rotate the plane of plane-polarized light. When describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of a molecule relative to its chiral center (s). The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane-polarized light by a compound, with (-) or l denoting that the compound is levorotatory. The connection with the prefix (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. A particular stereoisomer can also be called an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur in the absence of stereoselectivity or stereospecificity in a chemical reaction or process. The terms racemic mixture and racemate refer to an equimolar mixture of two enantiomeric molecules with no optical activity. Enantiomers can be isolated from a racemic mixture using chiral separation techniques such as supercritical liquid chromatography (SLC). The interpretation of the configuration at the chiral centers in the separated enantiomers can be preliminary, and it is shown in the structures of table. 1 for illustrative purposes, and finally the stereochemistry is established, for example, using X-ray crystallographic data.

The term tautomer or tautomeric form refers to structural isomers of different energies that are interconvertible across a low energy barrier. For example, proton

7,038835 tautomers (also known as prototropic tautomers) include interconversion via proton migration, eg keto-enol and imine-enamine isomerization. Valence tautomers involve interconversions by reorganizing some of the bonding electrons.

The term pharmaceutically acceptable salts refers to salts that are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid addition and base addition salts. The phrase “pharmaceutically acceptable” means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients contained in the composition and / or with the mammal to be treated.

The term pharmaceutically acceptable acid addition salt means such pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid mesylate, ethanesulfonic acid, p-toluenesulfonic acid and salicylic acid.

The term pharmaceutically acceptable base addition salt means such pharmaceutically acceptable salts formed with an organic or inorganic base. Examples of suitable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese and aluminum salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine resins, 2 -diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpamines.

The term solvate refers to an association or complex of one or more solvent molecules and a compound of this invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO (dimethyl sulfoxide), ethyl acetate (EtOAc), acetic acid (AcOH), and ethanolamine.

The term EC ₅₀ is half the maximum effective concentration and refers to the plasma concentration of a particular compound required to obtain 50% of the maximum in vivo of a particular effect.

The term Ki is an inhibition constant and denotes the absolute binding affinity of a particular inhibitor to a receptor. It is measured by competitive binding assays and is equal to the concentration at which a particular inhibitor occupies 50% of the receptors if there is no competing ligand (eg, a radiolabelled ligand). Ki values can be converted logarithmically to pKi values (-log Ki), where higher values indicate exponential increases in activity.

The term IC ₅₀ is half the maximum inhibitory concentration and refers to the concentration of a particular compound required to obtain 50% inhibition of an in vitro biological process. IC ₅₀ values can be converted logarithmically to pIC ₅₀ values (-log IC ₅₀ ), where higher values indicate exponential increases in activity. The IC50 value is not an absolute value, but depends on the experimental conditions, for example, the concentrations used, and can be converted to an absolute inhibition constant (Ki) using the Cheng-Prusov equation (Biochem. Pharmacol. (1973) 22: 3099). Other percent inhibition parameters such as IC ₇₀ , IC ₉₀ , etc. can be calculated.

The terms compound of this invention, compounds of this invention and compounds of Formula I include compounds of Formulas I and their stereoisomers, geometric isomers, tautomers, solvates, metabolites and pharmaceutically acceptable salts and prodrugs.

Any formula or structure set forth herein, including compounds of Formula I, is also used to represent hydrates, solvates, and polymorphs of such compounds, as well as mixtures thereof.

Any formula or structure recited herein, including compounds of Formula I, is also used to represent unlabeled forms as well as isotopically labeled forms of compounds. Isotopically labeled compounds have the structures depicted by the formulas set forth herein, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be included in

- 8 038835 compounds of this invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as, but not limited to, ² H (deuterium, D), ³ H (tritium), ⁿ C, ¹³ C , ¹⁴ C, ¹⁵ N, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I. Various isotopically labeled compounds of this invention, for example, those in which radioactive isotopes are incorporated such as ³ H, ¹³ C and ¹⁴ C. Such isotopically labeled compounds can be used in metabolic studies, kinetic response studies, detection or imaging techniques such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT ), including the analysis of the distribution of the drug or substrate in tissues, or in the radioactive treatment of patients. Deuterium-labeled or substituted therapeutic compounds of this invention may have improved drug metabolism and pharmacokinetics (DMPK) Absorption, Distribution, Metabolism, and Excretion (ADME) properties. Substitution with heavier isotopes such as deuterium, due to greater metabolic stability, may provide certain therapeutic benefits, such as increased in vivo half-life or reduced dosage requirements. ^{The 18} P-labeled compound can be used in PET or SPECT studies. Isotopically labeled compounds of this invention and prodrugs can generally be prepared by performing the procedures outlined in the schemes or in the examples and using the formulations described below, replacing the readily available isotopically labeled reagent with an unlabeled reagent. In addition, substitution with heavier isotopes, in particular deuterium (i.e., ² H or D), due to greater metabolic stability, can provide certain therapeutic benefits, for example, an increase in in vivo half-life or a decrease in dosage requirements or an improvement in the therapeutic index. It is understood that deuterium in this context is considered as a substituent on the compound of formula (I). The concentration of such a heavier isotope, in particular deuterium, can be determined using the isotopic enrichment factor. In the compounds of this invention, any atom that is not specifically designated as a particular isotope is intended to represent any stable isotope of that atom. Unless otherwise indicated, when a position is specifically defined as H or hydrogen, it should be understood that hydrogen is at that position in its natural isotopic composition. Accordingly, in the compound of the present invention, any atom specifically designated deuterium (D) is deuterium.

Estrogen receptor.

Estrogen receptor alpha (ER-α, NR3A1) and estrogen receptor beta (ER-β; NR3A2) are steroid hormone receptors that are members of the large nuclear receptor superfamily. Nuclear receptors share a common modular structure that, at a minimum, includes a DNA binding domain (DBD) and a ligand binding domain (LBD). Steroid hormone receptors are soluble intracellular proteins that act as ligand-regulated transcription factors. Vertebrates contain five closely related steroid hormone receptors (estrogen receptor, androgen receptor, progesterone receptor, glucocorticoid receptor, mineralocorticoid receptor) that regulate a wide range of reproductive, metabolic and developmental activities. ER activities are controlled by the binding of endogenous estrogens, including 17β-estradiol and estrone.

The ER-α gene is located at 6q25.1 and encodes a 595 amino acid protein. The ER-β gene is located on chromosome 14q23.3 and produces a 530 amino acid protein. However, due to the sites of alternative splicing and translation initiation, each of these genes can produce many isoforms. In addition to the DNA-binding domain (called the C-domain) and the ligand-binding domain (E-domain), these receptors contain an N-terminal (A / B) domain, a hinge (D) domain that links the C and E domains, the C-terminal elongation (F-domain) (Gronemeyer and Laudet; Protein Profile 2: 1173-1308, 1995). Although the C and E domains of ER-α and ER-β are fairly conserved (95% and 55% amino acid identity, respectively), the retention of domains A / B, D, and F is low (less than 30% amino acid identity). Both receptors are involved in the regulation and development of the female reproductive tract, and also play different roles in the metabolism of the central nervous system, cardiovascular system and bones.

The ligand-binding pocket of steroid hormone receptors is deeply embedded in the ligand-binding domain. After binding, the ligand becomes part of the hydrophobic core of this domain. Consequently, most steroid hormone receptors are unstable in the absence of the hormone and need assistance from chaperones such as Hsp90 to maintain hormone-binding competence. Interaction with Hsp90 also controls nuclear translocation of these receptors. Ligand binding stabilizes the receptor and initiates sequential conformational changes that release chaperones, alter interactions between different receptor domains, and remodel interaction surfaces with proteins, which allows these receptors to translocate into the nucleus, bind DNA, and participate in interactions with chromatin remodeling complexes and the transcriptional apparatus. Although the ER can interact with Hsp90, it is

- 9 038835 no interaction is required for hormone binding. and depending on the cellular context, ano-ER can be either cytoplasmic or nuclear. Biophysical studies have shown that DNA binding, rather than ligand binding, promotes receptor stability (Greenfield et al., Biochemistry 40: 6646-6652, 2001).

The ER can interact with DNA either directly by binding to a specific motif in the DNA sequence called the estrogen response element (ERE) (classical pathway), or indirectly through protein-protein interactions (non-classical pathway) (Welboren et al., Endocrinerelated Cancer 16: 1073- 1089, 2009). In a non-classical pathway, ER has been shown to bind to other transcription factors, including SP-1, AP-1, and NF-kB. These interactions appear to play an important role in the ER's ability to regulate cell proliferation and differentiation.

Both types of ER-DNA interactions can lead to gene activation or repression, depending on transcriptional co-regulators, which are recruited by the corresponding ERERE complex (Klinge, Steroid 65: 227-251, 2000). The attraction of coregulators is mainly mediated by two protein interaction surfaces, AF2 and AF1. AF2 is located in the E-domain of the ER, and its conformation is directly regulated by the ligand (Brzozowski et al., (1997) Nature 389: 753-758). Full agonists seem to promote the recruitment of coactivators, while weak agonists and antagonists facilitate the binding of corepressors. Protein regulation by AF1 is less understood but can be controlled by serine phosphorylation (Ward and Weigel, (2009) Biofactors 35: 528-536). One of the phosphorylation sites involved (S118) appears to control ER transcriptional activity in the presence of antagonists such as tamoxifen, which plays an important role in the treatment of breast cancer. While full agonists appear to block ER in a specific conformation, weak agonists seek to maintain the ER in equilibrium between different conformations, allowing cell-dependent differences in coregulator repertoires to modulate ER activity in a cell-dependent manner (Tamrazi et al., Mol Endocrinol 17: 2593-2602, 2003). ER-DNA interactions are dynamic and include, but are not limited to, degradation of the ER by the proteasome (Reid et al., Mol. Cell. 11: 695-707, 2003). Degradation of the ER by ligands provides an attractive strategy for treating diseases or conditions that are estrogen-sensitive and / or resistant to available anti-hormonal drugs. ER signaling is critical for the development and maintenance of the female reproductive organs, including the mammary gland, for ovulation and endometrial thickening. ER signaling also plays a role in the development of bone mass, lipid metabolism, cancer, etc. Approximately 70% of breast cancers express ER-a (ER-positive) and are estrogen-dependent for growth and survival. It is believed that the growth and survival of other cancers, such as, for example, ovarian and endometrial cancers, are also dependent on ER-α signaling. The ER-α antagonist tamoxifen has been used to treat early and advanced ER-a-positive breast cancer in pre- and post-menopausal women. Fulvestrant (FASLODEX®, AstraZeneca), a steroid-based ER antagonist, is used to treat breast cancer in women that progresses despite tamoxifen therapy (Howell A. (2006) Endocr Relat Cancer; 13: 689-706; US 6774122; US 7456160; US 8329680; US 8466139). Steroidal and non-steroidal aromatase inhibitors are also used in humans to treat cancer. In some embodiments, steroidal and non-steroidal aromatase inhibitors block the production of estrogen from androstenedione and testosterone in postmenopausal women, thereby blocking ER-dependent cancer growth. In addition to these antihormonal agents, progressive ER positive breast cancer is in some cases treated with a number of other chemotherapeutic agents, such as, for example, anthracilins, platinum compounds, taxanes. In some cases, ER-positive breast cancers that carry a genetic amplification of the ERB-B / HER2 tyrosine kinase receptor are treated with the monoclonal antibody trastuzumab (Herceptin®, Genentech Inc.) or the nixomolecular pan-ERB-B inhibitor lapatinib (TYKERB®, Glline .). Despite this battery of antihormonal, chemotherapy and small molecule drugs, and antibody-based targeting therapies, many women with ER-a-positive breast cancer develop progressive metastatic disease and need new treatment approaches. It is important to note that most ER-positive tumors that progress on existing antihormonal as well as other therapies appear to remain ER-a dependent for growth and survival. Thus, there is a need for new ER-a-targeted agents that are active in the setting of metastatic disease and acquired resistance. One aspect described herein are compounds that are selective estrogen receptor modulators (SERMs). In specific embodiments, the SERMs described herein are selective estrogen receptor degrader (SERD) agents. In some embodiments, in cellular assays, the compounds described herein result in a decrease in stable ER-a levels (i.e., ER degradation) and can be used to treat estrogen-sensitive diseases or conditions and / or diseases or conditions that have developed resistance to anti-hormonal therapy.

- 10 038835

Most breast cancer patients are treated with agents that either block estrogen synthesis (e.g., aromatase inhibitors; aromatase inhibitor, AI) or counteract the effects of estradiol through competitive ER binding (e.g. tamoxifen) (Puhalla S. et al. Mol. Oncol. 2012; 6 (2): 222-236). Despite the well-documented therapeutic benefits of these agents at various stages of the disease, many ER + breast cancers recur and eventually patients die. Recently, next-generation genome-wide sequencing and targeted sequencing have identified mutations in the ESR1 gene (estrogen receptor α gene) in 20% of tumors from patients with advanced breast cancer that has progressed on endocrine therapy, mainly on aromatase inhibitors (Li S., et al. Cell Rep. (2013); 4 (6): 1116-1130; Merenbakh Lamin K., et al. Cancer Res. (2013); 73 (23): 6856-6864; Robinson DR, et al. Nat. Genet (2013); 45 (12): 14461451; Toy W., et al. Nat. Genet. (2013); 45 (12): 1439-1445; Jeselsohn R., et al. Clin. Cancer Res. (2014 ); 20: 1757-1767). These mutations in the ligand-binding domain (LBD) give high basal activity of the apo-receptor, making it ligand-independent and thus active under low estradiol conditions. There is a need for a therapy targeting ER signaling with sustained activity in the setting of progressive disease after AI or tamoxifen treatment, including a subset of patients carrying ESRI mutant tumors.

In some embodiments, the compounds of Formula I described herein are used in methods of treating hormone-resistant estrogen (ER) -positive breast cancer in a patient characterized by a mutation in the ESR1 gene, said methods comprising administering a therapeutically effective amount of a compound of Formula I In some embodiments, a mutation in the ESR1 gene results in an ER polypeptide having an amino acid substitution at a position selected from amino acid positions 6, 118, 269, 311, 341, 350, 380, 392, 394, 433, 463, 503, 534, 535, 536, 537, 538 and 555 in SEQ ID NO: 2. In some embodiments, said mutation results in an ER polypeptide having an amino acid substitution selected from H6Y, S118P, R269C, T311M, S341L, A350E, E380Q, V392I, R394H , S433P, S463P, R503W, V534E, P535H, L536R, L536P, L536Q, Y537N, Y537C, Y537S, D538G and R555C. In some embodiments, the patient has two or more mutations in the ESR1 gene.

Given the central role of ER-α in the development and progression of breast cancer, the compounds described herein can be used to treat breast cancer alone or in combination with other agents that can modulate other critical pathways in breast cancer , including but not limited to those agents that target IGF1R, EGFR (Epidermal Growth Factor Receptor), CDK 4/6, erB-B2 and 3, PI3K / AKT / mTOR axis, HSP90, PARP (poly (ADP- ribose) -polymerase) or histone deacetylase.

Given the central role of ER-α in the development and progression of breast cancer, the compounds of formula I described herein can be used to treat breast cancer, either alone or in combination with another agent used to treat breast cancer, including , but not limited to, aromatase inhibitors, anthracyclines, platinum, alkylating agents, nitrogen mustards, taxanes. Illustrative agents used to treat breast cancer include, but are not limited to, PI3K inhibitors such as tasselizib (GDC-0032, Genentech Inc.), paclitaxel, anastrozole, exemestane, cyclophosphamide, epirubicin, fulvestrant, letrozole (FEMARA®, Novartis, Corp.), gemcitabine, trastuzumab, pegfilgrastim, filgrastim, tamoxifen, docetaxel, toremifene, vinorelbine, capecitabine (XELODA®, Roche), ixabepilone, and others described herein.

ER-related diseases or conditions include ER-α dysfunction associated with cancer (bone cancer, breast cancer, lung cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian and uterine cancer), central nervous system defects (alcoholism, migraine ), defects of the cardiovascular system (aortic aneurysm, risk of myocardial infarction, aortic valve sclerosis, cardiovascular disease, coronary artery disease, hypertension), defects of the hematological system (deep vein thrombosis), immune and inflammatory diseases (Graves' disease, arthritis, multiple sclerosis, cirrhosis), susceptibility to infection (hepatitis B, chronic liver disease), metabolic defects (bone density, cholestasis, hypospadias, obesity, osteoarthritis, osteopenia, osteoporosis), neurological defects (Alzheimer's disease, Parkinson's disease, migraine, dizziness ), mental defects (anorexia nervosa, attention deficit hyperactivity disorder (eng. attention deficit hyperactivity disorder, ADHD), dementia, major depressive disorder, psychosis) and reproductive defects (age of menarche, endometriosis, infertility).

In some embodiments, the compounds disclosed herein are used in the treatment of an estrogen receptor dependent or estrogen receptor mediated disease or condition in a mammal.

In some embodiments, the compounds described herein are used to treat cancer in a mammal. In some embodiments, the cancer is breast cancer, ovarian cancer,

- 11 038835 endometrial cancer, prostate cancer or uterine cancer. In some embodiments, the cancer is breast, lung, ovarian, endometrial, prostate, or uterine cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is hormone dependent cancer. In some embodiments, the cancer is estrogen receptor dependent cancer. In some embodiments, the cancer is estrogen sensitive cancer. In some embodiments, the cancer is resistant to anti-hormonal treatment. In some embodiments, the cancer is estrogen-sensitive cancer or estrogen receptor-dependent cancer that is resistant to anti-hormonal treatment. In some embodiments, the cancer is hormone sensitive or hormone receptor dependent cancer that is resistant to antihormonal treatment. In some embodiments, the anti-hormonal treatment comprises treatment with at least one agent selected from tamoxifen, fulvestrant, steroid aromatase inhibitors, and non-steroidal aromatase inhibitors.

In some embodiments, the compounds described herein are used to treat hormone receptor positive metastatic breast cancer in postmenopausal women with disease progression following antiestrogen therapy.

In some embodiments, the compounds described herein are used to treat a hormone-dependent benign or malignant breast or reproductive tract disease in a mammal. In some embodiments, the benign or malignant disease is breast cancer.

In some embodiments, the compound used in any of the methods described herein is an estrogen receptor degrading agent; an estrogen receptor antagonist; has minimal or negligible estrogen receptor agonist activity or a combination thereof.

In some embodiments, the methods of treating the compounds described herein include a treatment regimen that includes administering radiation therapy to a mammal.

In some embodiments, methods of treating the compounds described herein comprise administering the compound before or after surgery.

In some embodiments, methods of treating the compounds described herein comprise administering to the mammal at least one additional anti-cancer agent.

In some embodiments, the compounds described herein are used to treat cancer in a mammal where the mammal has not previously received chemotherapy.

In some embodiments, the compounds described herein are used in the treatment of cancer in a mammal. In some embodiments, the compounds described herein are used to treat cancer in a mammal, wherein the mammal is treated for cancer with at least one anti-cancer agent. In one embodiment, the cancer is hormone-resistant cancer.

In some embodiments, the compounds described herein are used in the treatment or prevention of diseases or conditions of the uterus in mammals. In some embodiments, the disease or condition of the uterus is a leiomyoma, uterine leiomyoma, endometrial hyperplasia, or endometriosis. In some embodiments, the disease or condition of the uterus is a cancer or condition of the uterus. In some other embodiments, the disease or condition of the uterus is a non-cancerous disease or condition of the uterus.

In some embodiments, the compounds described herein are used to treat endometriosis in mammals.

In some embodiments, the compounds described herein are used in the treatment of leiomyoma in a mammal. In some embodiments, the leiomyoma is uterine leiomyoma, esophageal leiomyoma, cutaneous leiomyoma, or small intestinal leiomyoma. In some embodiments, the compounds described herein are used to treat fibroids in a mammal. In some embodiments, the compounds described herein are used in the treatment of uterine fibroids in a mammal.

Another embodiment of this invention relates to a compound described herein for use as a therapeutically active substance.

Another embodiment of this invention relates to a compound described herein for use in the treatment of an ER-associated disease or disorder.

Another embodiment of this invention relates to the use of a compound described herein for the treatment of an ER-associated disease or disorder.

Another embodiment of this invention relates to the use of a compound described herein for the manufacture of a medicament that can be used to treat an ER-associated disease or disorder.

Tetrahydropyrido [3,4-b] indol-1-yl compounds

This invention provides tetrahydropyrido [3,4-b] indol-1-yl compounds of formula I, including formulas Ia-Ie, and pharmaceutical compositions thereof that can potentially be used for

- 12 038835 for the treatment of diseases, conditions and / or disorders modulated by the estrogen receptor α (ERa).

Compounds of formula I have the following structure:

pz \ / R ⁴

and its stereoisomers or pharmaceutically acceptable salts, where

Y ¹ represents CR ^b or N;

Y ² represents - (CH ₂ ) -;

Y ³ represents NR ^a or C (R ^b ) ₂ ;

where one of Y ¹ or Y ³ represents N or NR ^a ;

R ^{a is} independently H, C ₁ -C ₆ alkyl, C ₂ -C ₈ alkenyl, propargyl, C ₃ -C ₆ cycloalkyl or C ₃ -C ₆ heterocyclyl optionally substituted with one or more groups independently selected from F, Cl, Br, I, CN, OH, OCH3 and SO2CH3;

R ^b independently represents H, C ₁ -C ₆ -alkyl, C ₂ -C ₈ -alkenyl, propargyl, - (C ₁ -C ₆ -alkyldiyl) (C ₃ -C ₆ -cycloalkyl), C ₃ -C ₆ -cycloalkyl and C ₃ -C ₆ -heterocyclyl optionally substituted with one or more groups independently selected from F, Cl, Br, I, CN, —CH ₂ F, —CHF ₂ , —CF ₃ , —CH ₂ CF ₃ , —CH ₂ CHF ₂ , —CH2CH2F, OH, OCH3 and SO2CH3;

R ^c represents H;

Z ¹ represents a bond;

Cy is phenyldiyl or 5-6 membered heteroaryldiyl;

Z ² represents O or O- (C ₁ -C ₆ -alkyldiyl);

R ¹ and R ² independently represent H, -CH3, -CH2CH3, -CH (CH3) 2, -CH2CH (CH3) 2, -CH2OH, -CH2OCH3, -CH2CH2OH, -C (CH3) 2OH, -CH (OH ) CH (CH3) 2, -C (CH3) 2CH2OH, -CH2CH2SO2CH3, -CH ₂ OP (O) (OH) ₂ , -ch ₂ f, -chf ₂ , -ch ₂ nh ₂ , -ch ₂ nhso ₂ ch ₃ , —CH ₂ NHCH ₃ , —CH ₂ N (CH ₃ ) ₂ , —cf ₃ , —ch ₂ cf ₃ , —CH2CHF2, —CH (CH3) CN, —C (CH3) 2CN, or —CH2CN;

R ⁵ is H or CgC9 -alkyl, optionally substituted with one or more of the following substituents: halogen, CN, OR ^a , N (R ^a ) 2, C _g C ₉ -alkyl, C ₃ -C ₉ -cycloalkyl , C ₃ -C ₉ heterocycle, C6-C9 aryl, C (O) R ^b , C (O) NR ^a , SO2R ^a or SO2NR ^a ;

R ⁶ is F, Cl, Br, I, —CN, or —CH ₃ ; and m is 0 or 1;

where alkyldiyl, phenyldiyl and heteroaryldiyl are optionally substituted with one or more groups independently selected from F, Cl, Br, I, -CN, -CH3, -CH2CH3, -CH (CH3) 2, -CH2CH (CH3) 2 , -CH2OH, -CH2OCH3, -CH2CH2OH, -C (CH3) 2OH, -CH (OH) CH (CH3) 2, -C (CH3) 2CH2OH, -CH2CH2SO2CH3, -CH2OP (O) (OH) 2, -CH2F , -CHF2, -CF3, -CH2CF3, -CH2CHF2, -CH2CH2F, -CH (CH3) CN, -C (CH3) 2CN, -CH2CN, -CH2NH2, -CH2NHSO2CH3, -CH2NHCH3, -CH2N (CH3) 2, - CO2H, -COCH3, -CO2CH3, -CO2C (CH3) 3, -COCH (OH) CH3, -CONH2, -CONHCH3, -CON (CH3) 2, -C (CH3) 2CONH2, -NH2, -NHCH3, -N (CH3) 2, -NHCOCH3, -N (CH ₃ ) COCH ₃ , -NHS (O) ₂ CH ₃ , -N (CH ₃ ) C (CH ₃ ) ₂ CONH ₂ , -N (CH ₃ ) CH ₂ CH ₂ S (O) ₂ CH ₃ , -NO ₂ , = O, -OH, -OCH3, -OCH2CH3, -OCH2CH2OCH3, -OCH2CH2OH, -OCH2CH2N (CH3) 2, -OP (O) (OH) 2, -S (O) 2N (CH3) 2, -SCH ₃ , -S (O) ₂ CH ₃ , -S (O) ₃ H, cyclopropyl, cyclopropylamide, cyclobutyl, oxetanyl, azetidinyl, (1methylazetidin-3-yl) oxy, N -methyl-N-oxetan-3-ylamino, azetidin-1-ylmethyl, benzyloxyphenyl, pyrrolidin-1-yl, pyrrolidin-1-yl-methanone, piperazin-1- sludge, morpholinomethyl, morpholinomethanone and morpholino.

Compounds of formulas Ia-e have the following structures:

- 13 038835

where R ⁸ represents H or -CH3.

Illustrative embodiments of compounds of Formula I include those in which Y ¹ is CR ^b and Y ³ is NR ^a .

Illustrative embodiments of compounds of Formula I include those in which Y ¹ is N and Y ³ is C (R ^b ) 2.

Illustrative embodiments of compounds of Formula I include those in which Cy is phenyldiyl which is substituted with one or more F.

Illustrative embodiments of compounds of Formula I include those in which R ³ is H and R 4 is —CH 3.

Illustrative embodiments of compounds of Formula I include those in which R ⁵ is C ₁ -C ₆ fluoroalkyl.

Illustrative embodiments of compounds of Formula I include those in which m is 0.

Biological assessment

The relative activities of compounds of formula I as inhibitors of enzymatic activity (or other biological activity) can be determined by determining the concentrations at which each compound inhibits activity to a predetermined degree and then comparing these results. As a rule, it is preferable to determine the concentration that

- Paradise 14 038835 inhibits activity in biochemical analysis by 50%, i.e. determination of 50% inhibitory concentration, or IC ₅₀ . Determination of IC ₅₀ values can be accomplished using standard techniques known in the art. Typically, IC ₅₀ can be determined by measuring the activity of a given enzyme in the presence of a range of concentrations of the inhibitor of interest. The experimentally obtained values of enzyme activity are then plotted against the inhibitor concentrations used. The concentration of inhibitor that gives 50% enzymatic activity (compared to activity in the absence of any inhibitor) is taken as the IC ₅₀ value. Likewise, other inhibitory concentrations can be determined using appropriate activity determinations. For example, in some situations it may be desirable to determine 90% inhibitory concentration, i. E. IC ₉₀ etc.

Cell proliferation, cytotoxicity and cell viability under the influence of compounds of formula I can be measured using the CellTiterGlo® Luminescent Cell Viability Assay (Promega Corp.). This CellTiter-Glo® Luminescent Cell Viability Assay is a homogeneous method for determining the number of viable cells in culture by quantifying the presence of ATP, an indicator of metabolically active cells. The CellTiter-Glo® assay is designed for use in multi-well formats, making it ideal for automated highthroughput screening (HTS), cell proliferation and cytotoxicity assays. The homogeneous assay procedure involves the addition of a single reagent (CellTiter-Glo® Reagent) directly to cells cultured in medium supplemented with serum. Cell washing, media removal, and multiple pipetting steps are not required. The system detects a total of 15 cells / well in 384 well format 10 min after reagent addition and mixing.

All illustrative compounds of formula I in table. 1 and 2 were prepared and characterized by LCMS [M + H] + (liquid chromatography with mass spectroscopy) with detection of the precursor ion. All illustrative compounds of formula I in table. 1 and 2 were tested for binding to ERa (estrogen receptor α) and for biological activity in accordance with the assays, protocols and procedures described in examples 901-907. ER-α MCF7 HCS S _inf values (%) in table. 1 was measured using a high-precision analysis of degradation based on fluorescence imaging ERa in breast cancer cells (Breast Cancer Cell ERa High Content Fluorescence Imaging Degradation Assay) described in example 901. The values of ER-α MCF7 HCS EC ₅₀ (μM) in table. 1 and 2 were measured using the in vitro cell proliferation assays described in examples 902 and 903. The rat uterus gross weight assays described in examples 906 and 907 quickly determine the antagonistic activity of compounds in tissue responsive to ER (immature rat uterus), when competing with the native ER ligand estradiol, i.e. in antagonist mode (Ashby, J .; et al. (1997) Regulatory toxicology and pharmacology: RTP, 25 (3): 226-31). Illustrative compounds of formula I in table. 1 and 2 have the following structures, corresponding names (ChemBioDraw, version 12.0.2, and CambridgeSoft Corp., Cambridge, MA, USA) and biological activity. If more than one name is associated with a compound of Formula I or an intermediate, the chemical structure defines the compound.

- 15 038835

Table 1

No. Structure Name ERalpha MCF7 HCS ec ₅₀ (μM) ERalpha MCF7 HCS s _inf (%) 101 w ZI (1R, 3R) -1- (2,6 difluoro-4 - ((1- (3fluoropropyl) azeti din-3- yl) oxy) phenyl) -2 (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.000049 -one hundred 102 _f \ -F JN— 'ΓχΜ, / ^H f4 / / \ \ = ς α-ν y— _h F (1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 2- (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.000014 -98.9% 103 o n) —Ζ ^F Μ O— \ / \ V — Ν> - \ F 1 - ((1R, 3R) -1- (2.6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1Hpyrido [3,4 - B] indol-2 (9H) -yl) 2-methylpropan-1one 0.00011 -91.8%

- 16 038835

104 F Г nA ΓχΟ ° O ^ N г F H; - / ^F ^ / \ F Z \ J v ^ n> - 1 - ((1 R, 3R) -1 - (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1Hpyrido [3, 4- B] indol-2 (9H) -yl) 2-fluoro-2methylpropan-1-one 0.000043 -95.4% 105 / \ ^F О'-i / --Х F ^F O Of О (1R, 3R) -1 - (4- (2- (3 (difluoromethyl) azetidin-1yl) ethoxy) -2,6difluorophenyl) -2 (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro -1Hpyrido [3.4b] indole 0.000027 -94% 106 ... F About Ν Ο O / O. / h 05 ^ Ov. Cl (1R, 3R) -1 - (4 - ((1- (3chloropropyl) azetidin-3-yl) oxy) -2.6difluorophenyl) -2 (2-fluoro-2methylpropyl) -3methyl-2,3,4, 9tetrahydro-1Hpyrido [3.4b] indole 0.000142 -90.4% 107 50 O 'ZI (1R, 3R) -1- (2,6difluoro-4 - ((1propylazetidin-3yl) oxy) phenyl) -2 (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b ] indole 0.000024 -97.6% 108 ... F A nO 0 / 0.0 zrA ^Hf 4 = J Ο-l Λ ~ Ί / ⁿ ^ o ^ ^f (1R, 3R) -1- (2,6 difluoro-4 - ((5) -2 ((R) -3 (fluoromethyl) pyrrolidin-1yl) propoxy) phenyl) -2- (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.00011 -79.3%

- 17 038835

109 _χ F jy N— ⁷ ГД ^H FO O— \ / --- 1 (1R, 3R) -1- (2,6 difluoro-4- (2 - ((I) 3- (fluoromethyl) pyrro lidine-1- yl) ethoxy) phenyl) 2- (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.000050 -86.5% 110 111 __f yF N— ⁷ (XV4 V ²⁴ N / TA H (/ N O— \ A__ VN> - \ VF (1 R, ЗR) -2- (2-φτop2-methylpropyl) -1 (6- (2- (3- (fluoromethyl) azeti din-1- yl) ethoxy) pyridin-3-yl) -3-methyl- 2,3,4,9- tetrahydro-1Hpyrido [3.4b] indole 0.000898 -96.8 112 \ F 14- ⁷ CHAIR / ^v %, ° Ά-ν (> - ¹ (1S, ЗR) -2- (2-φτop2-methylpropyl) -1 (3-fluoro-5- (2- (3 (fluoromethyl) azeti din-1- yl) ethoxy) pyridin -2-yl) -3-methyl 2,3,4,9- tetrahydro-1Hpyrido [3.4b] indole 0.000092 -93.9 113 si— (Tv, r ^N Xa ^N ChE r ^ F Sq / X / NJ (1R, 3R) -2- (cyclobutylmethyl) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 3-methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.00618 -94.8

- 18 038835

114 r ° Ν— ⁷ (Гн / ίτΑ ^f O гУ (1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-2 - ((3methyloxetane-3yl) methyl) -2 , 3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.000715 -97.8 115 NV Cm.j ^ N 'r- \ ^H CHE ^ F (1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-2 (oxetane-3ylmethyl) -2,3,4 , 9tetrahydro-1Hpyrido [3.4b] indole 0.0231 -95.5 116 I— \ C ГУч / x ~ ' ^N W. ^H 4J r ^ F (1R, 3R) -1- (2,6 difluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-2 (oxetan-3-yl) 2,3,4, 9tetrahydro-1Hpyrido [3.4b] indole 0.0414 -85 117 o— ¹ (1R, 3R) -1- (2,6d and fluoro-4- (2 (pyrrolidin-1yl) ethoxy) phenyl) 2- (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [ 3.4b] indole 0.000171 -71.7 118 (TUch / ^F X_ (λΌ (1R, 3R) -1- (2,6difluoro-4- (2 (piperidin-1yl) ethoxy) phenyl) 2- (2-fluoro-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3, 4b] indole 0.000488 -72.2

- 19 038835

119 * \ ^F n— Г iPM 7 OH ^{H f} a / (1 - (2- (3,5-difluoro4 - ((1 R, 3R) -2- (2φτορ-2methylpropyl) -3methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indol-1yl) phenoxy ) ethyl) and zetidine-3yl) methanol 0.00198 -71.3 120 IZ ν '^ z ^^ Ο Ή I Ή v (1 R, ЗR) -2- (2-φτop2-methylpropyl) -1 (2-fluoro-4- (2- (3 (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 3-methyl-2,3,4,9tetrahydro-1Hpyrido [3,4b] indole 0.000056 2 -96.3 121 _ [o ΐΟΤζ- ^ ^f n / == 4 ^F V-D ^ O- ^/ F 1 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1Hpyrido [3,4b ] indol-2 (9H) yl) ethanone 0.008 -91 122 / 0 (TH _F ^{0H Hp} ^ О- / xX-A 1 - ((1R, 3R) -1- (2.6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1H-pyrido [3,4- B] indol-2 (9H) -yl) 2-hydroxy-2methylpropan-1-one 0.0014 -97.8 123 _f <OH N— (ΓΗ, ^F % = < ^H V_2Z ^ N Y— \ O- ⁷ F (R) -3 - ((1R, 3R) -1 (2,6-difluoro-4- (2 (3- (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1H-pyrido [3,4- B] indol-2 (9H) -yl) 2-methylpropan-1ol 0.00104 -96.1

- 20 038835

124 OH Cgm ° _F Y ^ -N g / = J ^F V / Λ \ / Ν \ __. ο-χ \ F ((1 R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-3,4-dihydro-1Hpyrido [3,4b] indole-2 (9Н) nn) ((1s, 3S) -3hydroxycyclobut yl) methanone 0.135 -75 125 shua and UA ^F Μ Ο-U ^F 1 - (1- (2,6-difluoro4- (2- (3- (fluoromethyl) azeti din-1- yl) ethoxy) phenyl) 3,3-dimethyl-3,4-dihydro-1H-pyrido [3,4- B] indol-2 (9H) -yl) 2-methylpropan-1one 0.0744 -90 126 W iz TI j I 11 <p About 1Ό (1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azeti din-1yl) ethoxy) phenyl) 3-methyl-2 (methylsulfonyl) -2,3,4,9tetrahydro -1Hpyrido [3.4b] indole 0.000469 -94.4

table 2

No. Structure Name ERalpha MCF7 HCS ec ₅₀ (μM) LCMS [M + H] ⁺ 127 About ^zz I Jr-A about 1 - [(1R, 3R) -1- [2,6d and fluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4,9tetrahydropyrido [3 , 4b] indol-2-yl] -2methylpropan-2-ol 0.000641 502.6

- 21 038835

128 .0 F ^ T-J F (1R, 3R) -1- [4- [2- [3- (difluoromethyl) azetidin1-yl] ethoxy] -2,6-difluorophenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000064 522.3 129 _Γ \ -F ΓΧ><Ζ / GA ^nr CHE ο CI— (1R, 3R) -1- [4- [1- (3- chporpropyl) azetidine-3yl] oxy-2,6-difluorophenyl] -2- (2-fluoro-2methyl-propyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.00017 522.2 130 _Λ D-F Comp J Ν ^ -4 ^H F ^ O Ο (1Р, ЗР) -1- [2,6-difluoro-4 (1-propylazetidin-3yl) oxy-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000050 4 486.3 131 ^ΗΟ \ CrtZ / ^N “^ ^Η -Ο. F '- (y — Ν 2— / ο— / ((18, ЗР) -1- (2,6-difluoro4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2- (2-fluoro-2-methylpropyl) 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indol-3yl) methanol 0.000146 520.3

- 22 038835

132 _F \ -F Ν— ' O,. Vnh (1 R, 3R) -1 - [4- (azetidin3-yloxy) -2,6-difluorophenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.00031 444.2 133 0 0 --N F— ' (1Р, ЗР) -2- (2-fluoro-2methylpropyl) -1 - (2fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.000056 2 486.3 134 O IZ I jyvl (1 H, ZR) -2-cyclobutyl-1 [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000542 484.3 135 F CWZ / ^ ^H / 1 ^F \ / / \ ^F 'x / - ^Z o-E (Sh, 35) -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetidin-1 yl] ethoxy] phenyl] -3- (fluoromethyl) -2- (2-fluoro-2-methyl-propyl) -1,3,4,9 tetrahydropyrido [3,4b] indole 0.000097 3 522.3 136 > i— ° / \ ^f 4_I VFL / ^ N i F Η ί— / ^Ρ -θ О F (1K, ZR) -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2 - [(3fluoroxetane-3- yl) methyl] -3-methyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.000114 518.3

- 23 038835

137 O \ P κν ° 1 ^ζχ cyclohexyl ((1 R, 3R) -1 (2,6-d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9H) -yl) methanone 0.028 540.4 138 suU Ν ί 0 ⁿ Μ ^F ~ Q 1 - [(1R, 3R) -1- [2.6- d and fluoro p-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indol-2-yl] -2.2- dimethylpropan-1-one 0.000491 514.3 139 \ C ⁿ ~~ v ^ Ν ί О ^H K- ^ ^F cyclopropyl ((1 R, 3R) -1 (2,6-d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9H) -yl) methanone 0.00248 498.2 140 __χ ~~~ Ρ \ F \ -p N 3 F H ^F ^ QO (1Р, ЗР) -1- [2,6-difluoro-4 [2- (3-methylazetidin-1 yl) ethoxy] phenyl] -2- (2fluoro-2-methyl-propyl) 3-methyl-1,3 , 4.9 tetrahydropyrido [3.4b] indole 0.00013 486.4 141 __ __P \ F \ P N- ~ y ^ N 'F H> == y ^F p) v-Cr (1Р, ЗР) -1- (2,6-difluoro-4 (2 - ((R) -3methylpyrrolidin-1yl) ethoxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4 ,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000151 500.2 142 __ __A \ F N i F ^f O * X-PO (1Р, ЗР) -1- [2,6-difluoro-4 [(28) -2-pyrrolide in-1 ylpropoxy] phenyl] -2- (2fluoro-2-methyl-propyl) 3-methyl-1,3 , 4.9 tetrahydropyrido [3.4b] indole 0.000090 500.3

- 24 038835

143 ____ Λ \ F C ⁿ - ^np b < ^F N— ¹ o— / ^/ (1I, ZK) -1- [2,6-difluoro-4- [3- [3- (fluoromethyl) azetid yn-1 yl] propoxy] phenyl] -2- (2-fluoro-2-methylpropyl) -3-methyl-1,3,4,9tetrahydropyrido [3,4- B] indole 0.000078 518.4 144 y - f \ F N i _P H XV ^F ^ O F (1K, ZI) -1- [2,6-difluoro-4- [(E) -3- [3- (fluoromethyl) azetid in-1 yl] prop-1-eny1] phenyl] -2- (2-fluoro-2-methyl- propyl) -3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000144 500.1 145 y - f \ F N i F H ^F ~ w - F ΗΝ-Αλ z— / M- (3,5-difluoro-4- ((1K, ZI) -2- (2-fluoro-2methylpropyl) -3-methyl- 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indol-1 yl) phenyl) -1- (3- fluoropropyl) azetidine-3amine 0.000096 503.3 146 F f-l-F N 'F H y __ / °> F (1R, 3R) -2 - [(3,3- difluorocyclobutyl) methyl] -1- [2,6-difluoro-4- [2- [3 (fluoromethyl) azetidin-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.01 534.3 147 N i FH ^F <3 ° A F (1R, ЗR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2,2dimethylpropyl) -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.000698 500.3

- 25 038835

148 Ν - 0 ^{Η F} cyclobutyl - [(1 R, 3R) -1 [2,6-difluoro-4- [2- [3 (fluoromethyl) azetidine-1yl] ethoxy] phenyl] -3- methyl 1,3,4,9tetrahydropyrido [3,4b] indol-2-yl] methanone 0.00147 512.3 149 sshV N -, 0 ^H F ^ r ^F cyclopentyl - [(1 R, 3R) -1 [2,6-difluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl 1,3,4,9tetrahydropyrido [3,4b] indol-2-yl] methanone 0.00216 526.4 150 __. __ £ \ F N i F ^f ~ Q (1Н, ЗИ) -1- [2,6-difluoro-4 [2 - [(3S) -3- methylpyrrolidine-1yl] ethoxy] phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000483 500.2 151 __ __X \ FN ^ Z N 'F H ^F ^ Q <° ^ ^N O (1H, 3I) -1- [2,6-difluoro-4 [(2H) -2-pyrrolidin-1 ylpropoxy] phenyl] -2- (2fluoro-2-methyl-propyl) 3-methyl-1,3, 4.9 tetrahydropyrido [3.4b] indole 0.000156 500.1 152 __ __ £ \ F \ - £ N> F H / ΓΪ p-U (III, ZI) -1- [2,6-difluoro-4 [(1-propylazetide in-3yl) methoxy] phenyl] -2- (2fluoro-2-methyl-propyl) 3-methyl-1,3,4 , 9tetrahydropyrido [3.4b] indole 0.000362 500.3

- 26 038835

153 __> F dDDm-yO N i F Η ^F ° ~~ l F (1I, 3N) -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2 - [(1 fluorocyclobutyl) methyl] 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000221 516.3 154 > F / - \ \ / C ⁿ - H Λ— / ^F ^ O o f (S) -3 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro -1 Npyrido [3,4-b] indol 2 (9H) -yl) -2-fluoro-2methylpropan-1-ol 0.000033 3 520.3 155 F OH r thA — J Г iJVi.j ^Η ^ θ О N X (2R) -3 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2-fluoro-2methylpropan-1-ol 0.000079 9 520.3 156 _Λ \ d h 0 N F ^ (1R, 3R) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000335 468.3

- 27 038835

157 О ιζ] sТ 2-cyclopropyl-1 [(1R, ЗR) -1- [2,6-difτop- 4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] ethanone 0.005 512.4 158 \ Y v ^N - ⁰ н г f ^F ~ w ^ 0-Α ^ ^Ν ζΛ- ~ / ^Ρ 2-cyclobutyl-1 - [(1 R, 3R) 1- [2,6-difluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4 , 9tetrahydropyrido [3,4b] indol-2-yl] ethanone 0.01 526.4 159 ΟίΑ Ν 1 0 H g- F h ° Ά ^ ^Ν ζΛ ~ - / ^Ρ 1 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetidin-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2,2 difluoro-propan-1-one 0.000147 522.3 160 ____ Λ \ F) Τ Ν ~ - / ^ Ν i. F Η ζ == - Ζ ^F ^ Q η ^υ - V ^ \ F (1R, ZR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) -3methyl-azetidin-1 yl] ethoxy] phenyl] -2- (2fluoro-2-methylpropyl) 3 -methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000285 518.4 161 Ο ^ιζ A Ad 1 (1R, 3R) -1- [2,6-difτop-4 (1-methylazetidine-3yl) oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4- B] indole 0.000041 3 458.3

- 28 038835

162 oyu n ^ 7 == ^ F ^ / \ V / (1R, 3R) -1- [4- (1- ethylazetidin-3-yl) oxy2,6-difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000052 eight 472.2 163 SNL ⁴ N i F n y = ~ z ^F ~ O o_ / A .— / (III, 3I) -1- [2,6-difluoro-4 (1-pentylazetidin-3yl) oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000457 513.9 164 / \ F A / H 7 = ( F ^ / \ V / (1R, 3R) -1- [4- [1- (cyclopropylmethyl) azet idin-3-yl] oxy-2,6difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000216 498.4 165 __Z \ F ^N b 7 h 7 = (f— \ νγ / k (1R, 3R) -1- [4- [1- (cyclopentylmethyl) azet idin-3-yl] oxy-2,6difluoro-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.002 526.5 166 / ~ - ^ \ ^f \ / Al n ^ - '> 7 n 7 = (F ^ Z \ V / (1H, 3H) -1- [2,6-difluoro-4 [1- (2- fluoroethyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000175 490.4 167 f \ F N 7, f ^H F ^ M (1H, 3H) -1- [2,6-difluoro-4 (1-prop-2-ynylazetidin3-yl) oxy-phenyl] -2- (2fluoro-2-methyl-propyl) 3-methyl-1,3 , 4.9 tetrahydropyrido [3.4b] indole 0.000083 482.3

- 29 038835

168 f \ F N к / н / = O FV \ ° ^ _Ν γ (L, 3 ^ -1- [2,6-difluoro-4 (1-isopropylazetidin-3yl) oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000122 486.4 169 ___A \ F \ to-y h 7 == ( F ^ Z \ (1K, ZI) -1- [2,6-difluoro-4 (1-isobutylazetidin-3yl) oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000409 400.3 170 χ \ / - \ F ^ / N 9 F H Тт-У Ο ^ λ o ° o tert-butyl-3- [3,5- difluoro-4 - [(1K, 3I) -2- (2-fluoro-2-methyl-propyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole-1- yl] phenoxy] azetidine-1 carboxylate 0.001 544.3 171 v / l / Op H ^F '^ C /' o-Y-X-Y (1K, ZI) -1- (2,6-difluoro-4- (2- (3- (fluoromethyl) azetid yn-1 yl) ethoxy) phenyl) -3- ethyl 2- (2-fluoro-2methylpropyl) -2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000471 518.3 172 __ 5 v / of? ^N ^^ ^F and O / ^{F f} ~ Y) bv ^ ^N 0o- / ^F (1I, 38) -1- (2,6-difluoro-4- (2- (3- (fluoromethyl) azetid yn-1 yl) ethoxy) phenyl) -3- ethyl 2- (2-fluoro-2methylpropyl) -2,3,4,9tetra hydro-1 Npyrido [3,4-b] indole 0.017 518.3

- 30 038835

173 N i F H ^F ^ F (1I, ZK) -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-2 - [(1- methylcyclobutyl) methyl ] -1,3,4,9- tetrahydropyrido [3.4b] indole 0.001 512.3 174 _ / - / V N '- F H / ^F V? ° A F (Ш, ЗР) -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-2- (2,2,2trifluoroethyl) -1,3,4,9tetrahydropyrido [3,4b] indole 0.000285 512.2 175 / { ^F \ / ^F Ox / - / ^f N ^ 'VF H Y — Y ^F v3 ° T F (18.3 ^ -1- [2,6-difluoro-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-2- (2,2,2trifluoroethyl) -1,3,4,9tetrahydropyrido [3,4b] indole 0.014 512.2 176 О TZ. Д ^ Ж ζ 0 ^ 0 I I (1R, 3R) -1- [4- [1- (3,3dimethoxypropyl) azeti din-3-yl] oxy-2,6difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl- 1,3,4,9tetrahydropyrido [3,4b] indole 0.000031 546.3 177 F \ —F N-Z - ό o— ^ n— F (1 R, ЗR) -1- [2-φτop-4- [1 (3- phtopropyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methyl-propyl) 3-methyl-1,3,4,9 tetrahydropyrido [3,4b] indole 0.000164 486.3

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178 \ —F Л N— 'OYO ^Hf -O 1- [2,6-difluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2fluoro-2-methylpropyl) 1,3,4,9- tetrahydropyrido [3.4b] indole 0.0005 490.2 179 Λ 0 J \ r ^s * (th ° θ о Ν (1S, 3R) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-2- methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.013 472.2 180 Л О / - \ С N ^ A Сч ° Q About N (1 R, 3R) -1 - (4- (2- (3- (fluoromethyl) azetid yn-1 yl) ethoxy) phenyl) -3- methyl-2- (methylsulfonyl) - 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.000222 472.2 181 Suchu the t N o ⁿ ί F 1 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [ 3,4-b] indol2 (9H) -yl) -3-hydroxy-2methylpropan-1-one 0.004 516.2 182 __> I— NH \ Y d ^N T \ N i 0 HF ^f ~ o 'o. ^' Y-Y azetidin-3-yl - [(1 R, 3R) 1- [2,6-difluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4 , 9tetrahydropyrido [3,4b] indol-2-yl] methanone more than 0.1 513.3

- 32 038835

183 chHl ν g ^{n f} ~ O ((1I, ZK) -1- (2,6-difluoro4- (2- (3- (fluoromethyl) azetid yn-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9H) -yl) (2- fluorocyclopropyl) methane he 0.004 516.3 184 __ __Λ \ F Ν / Ν 1 F Η ζΑΑ ^f O * 4> Ο (1I, ZK) -1- [2,6-difluoro-4 [(2S) -2 - [(3R) -3- methylpyrrolidine-1yl] propoxy] phenyl] -2 (2-fluoro-2-methylpropyl) -3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000494 514.4 185 oH ⁰ ng f ^F ~ w [(1Р, ЗР) -1- [2,6-difluoro4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] phenylmethanone 0.014 534.2 186 Ν b Η £ F ° ~ Λ ^Vn ^ ^F (1R, 3R) -2- (cyclopropylmethyl) -1 [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid yn-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.002 484.4 187 f \ F f N ~ - / ' ^N > / ^F (1R, 3R) -1- [4- [1- (2cyclopropylethyl) azetid in-3-yl] oxy-2,6difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl-1, 3,4,9tetrahydropyrido [3,4b] indole 0.001 512.4

- 33 038835

188 (1R, 3R) -1- [4- (1- allylazetidin-3yl) oxy-2,6-difluorophenyl] -2- (2-fluoro-2methyl-propyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000128 484.3 189 f \ F Г / N - 7 ~ ^{H f} ^ 4 ⁼ XV / (1R, 3R) -1- [4- [1- (cyclobutylmethyl) azeti din-3-yl] oxy-2,6difluoro-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.001 512.3 190 \ ^F Г N g., ^F h 7 = / F— / \ V / (1Р, ЗР) -1- [2,6-difluoro-4 (1-isopentylazetidin-3yl) oxy-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.002 514.3 191 O IZ I z ^^ (1Р, ЗР) -1- (2,6-difluoro-4 ((1- (2- methylbutyl) azetidine-3yl) oxy) phenyl) -2- (2- fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.000267 514.3 192 f \ F 7 ^{,, F H} f-JM νγ ° χ \ ^ (1Р, ЗР) -1- (2,6-difluoro-4 ((1- (pentane-2- yl) azetidine-3yl) oxy) phenyl) -2- (2-fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.001 514.3 193 ^zx 0 (1R, 3R) -1- [4- (1- cyclobutylazetidin-3yl) oxy-2,6-difluorophenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000484 498.3

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194 O ΞΕΖ J oil (1I, ZK) -1- [2,6-difluoro-4- [1- (oxetane-3- yl) azetidin-3-yl] oxyphenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000077 500.2 195 M \ F J N — 7 (Thj (1R, 3R) -1- [4- (1- cyclopropylazetidine-3yl) oxy-2,6-difluorophenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000165 484.3 196 f \ F \ / V A- '' η \ f H Ad F-ν \ V / - <F (1R, ЗR) -1- [2,6-diφτop-4 [1- (3- fluoropropyl) azetidine-3yl] sulfanyl-phenyl] -2 (2-fluoro-2-methylpropyl) -3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000271 520.3 197 N i F H 5 = / ^F ^ O F (1R, ЗR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2isobutyl-3-methyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.001 486.2 198 N i F H 5 == / ^F ^ Q F (1R, ЗR) -1- (2,6-diφτop-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-2- (1) -2phenylpropyl) -2,3,4,9tetrahydro-1 Npyrido [3,4-b] indole 0.002 548.1

- 35 038835

199 N i FH ^F ~ w F (1I, ZK) -1- (2,6-difluoro-4- (2- (3- (fluoromethyl) azetid yn-1 yl) ethoxy) phenyl) -3methyl-2 - ((8) -2- phenylpropyl) -2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.00232 548.3 200 d ^N - / N i F H аУ Q Ο -— Ά (1I, ZK) -1- [2,6-difluoro-4 (1-propylazetidin-3yl) oxy-phenyl] -2isobutyl-3-methyl 1,3,4,9 tetrahydropyrido [3,4b] indole 0.000408 468.3 201 OTUL N - H / <, О 0 N—। F ^ (1K, 3I) -2- (2-fluoro-2methyl-propyl) -1 - [4- [1 - (3fluoropropyl) azetidine-3yl] oxyphenyl] -3-methyl1,3,4,9tetrahydropyrido [3,4b] indole 0.000315 202 £ 0 η /. t \ u (Tuch / ^nr LE O. F (1K, ZI) -1- [2,6-difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl2-methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.0001 508.2

- 36 038835

203 F ° H i F \ Ζ ^ νΛ / Ο-U - / [1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (3fluorocyclobutyl) methano n 0.004 530.4 204 ____P \ F \ PCN ^ / ^ N i F H ^F ~ w F (1R, 3R) -1- [4 - [(2S) -2- [3 (difluoromethyl) azetidin1-yl] propoxy] -2,6difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3- methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000371 536.4 205 __. / -— P \ F N b F H ^F ^ Q n ^N ^ F ^F (1Р, ЗР) -1- [2,6-difluoro-4 [2- [3-fluoro-3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000375 522.1 206 N ^{Ί 1} Г / Λ> - / о n г and г f ^F 4j 4 - [(1R, 3R) -1- [2.6- d and fluoro p-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -4-oxobutanenitrile 0.031 511.3 207 cn5% Ν; n g F h ° ~ l (1R, 3R) -2- (cyclohexylmethyl) -1 [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.013 526.4

- 37 038835

208 f \ F ^H 4 ^ 0 ° (1I, ZK) -1- [2,6-difluoro-4 [1- [2- (oxetane-3yl) ethyl] azetidine-3yl] oxy-phenyl] -2- (2fluoro-2-methyl-propyl) 3 -methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000066 528.3 209 N i F H Dd d (1R, 3R) -1- [4- [1- (cyclohexylmethyl) azet idin-3-yl] oxy-2,6difluoro-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.008 540.3 210 \ F __ / N— ⁷ 0 < ^N F ^ (1R, 3R) -1- [2-xnop-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2-fluoro-2-methyl-propyl) 3-methyl-1 , 3,4,9tetrahydropyrido [3,4b] indole 0.0001 503.2 211 ... F Τ 'N—' ^ ¹ F (1R, 3R) -1- [2-xnop-4- [1 (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.0002 503.2 212 f OH \ 7 V ^N —Z ' ⁰ h g f ^F ~ w 1 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [ 3,4-b] indole- 2 (9H) -yl) -3- hydroxybutane-1-one 0.001 538.2

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213 ° —1 / v / XY ° ^ZI О [(1I, ZK) -1- [2,6-difluoro4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (oxetan3-yl) methanone 0.004 514.2 214 __> 1 — s d ^N ~ d O H + F ^F ^ Q o ^ ⁿ Q ^ ^F [(1K, ZI) -1- [2,6-difluoro4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (thietan-3yl) methanone 0.004 530.2 215 Ιζ ^ ί ^ j -P ? ' (R) -1 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4 , 9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -3-fluoro-2methylpropan-1-one 0.000335 518.3 216 \ A d ^N —X N i n g f 4 o (1R, 3R) -2- (cyclopentylmethyl) -1 [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.001 512.4 217 qA + GH ^F ο-Ά ^ z— / Λ ^ ^Ν -> Ζ (1R, 3R) -1- [4- [1 - [(4,4difluorocyclohexyl) methyl] azetidin-3-yl] oxy2,6-difluoro-phenyl] -2- (2fluoro-2-methyl-propyl) 3 -methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.002 576.3

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218 O ^vz 1 o ^ nc o z} _{> d} \ (S) -1 - ((1 R, 3R) -1 - (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3, 4.9 tetrahydro-2Hpyrido [3,4-b] indol-2yl) -3-fluoro-2methylpropan-1-one 0.000402 518.3 219 o IZ] / Γ '* z \ o ^ Chok J / - ^Χ -ηΟ ^ \ / o-l ((1R, ЗR) -1- (2,6-dφτop4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-L] indol-2yl) (oxetane-2yl) methanone 0.035 514.2 220 Oh NH Seyu / Y1EL O— \ / k '-' ^N / - \ Yu F (1R, ЗR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.007 430.2 221 No.h ° - Q about N 2 ^ Top-1 - [(1R, 3R) -1- [4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4- B] indol-2-yl] -2methylpropan-1-one 0.18 482.3 222 Y P ° A X j ^ γ ^ ΖΙ 1 - [(1R, 3R) -1- [2.6- difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2-fluoro-2methylpropan-1-one 0.0004 518.2

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223 Ν ι О ^н ί F ^F ~ O ο-> Ά-Χ ^Ρ 1 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2- (dimethylamino) ethanone 0.038 515.2 224 N i F H?: __ / ^Ρ Ο о ~ - ^ А X F (1H, 3H) -1- [2,6-difluoro-4 [1 - [(1- fluorocyclopropyl) methyl] azetidin-3-yl] oxyphenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000316 516.2 225 n ^z r o ^H ZF X o - ^ A F [(1H, 3H) -1- [2,6-difluoro- 4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (1fluorocyclobutyl) methano n 0.001 530.1 226 n ^z g about n b F ^F h o ^ / A F [(1H, 3H) -1- [2,6-difluoro- 4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (1methylcyclopropyl) methanone 0.002 512.1

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227 / - ζ ΛΊ F N i F H Ay O- ~ / \ F (1I, ZK) -1- [2,6-difluoro-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2 - [[1 - (fluoromethyl) cyclopropyl] methyl] -3-methyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.00017 516.1 228 OvW ^Hf -Q F [1 - [[(1R, 3R) -1- [2.6- difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4- B] indol-2yl] methyl] cyclopropyl] methanol 0.001 514.2 229 / \ ^F / nA CyW! ^{H f} O 0 N X 2-φτop-1 - [(1S, ЗR) -1- [2fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4,9tetrahydropyrido [3,4b] indol-2-yl] -2methylpropan-1-one 0.025 500.2 230 IZ jij O ^ Ai ... / —C> / z— ' 2-fluoro-1 - [(1 R, 3R) -1- [2fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9tetrahydropyrido [3,4b] indol-2-yl] -2methylpropan-1-one 0.000068 500.2

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231 Lo, --- \ fTVC ° HA ^{Η f} Ό Ο Ν (18, ZI) -1- [2-fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-2- methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.024 490.2 232 ιζ ίι \ -θ Y Λ, -οί ^ ο ^ζ \ / - \ ^ζ - (1K, ZI) -1- [2-fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-2- methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.000192 490.2 233 y - \ \ ^ ^0Η Ν> F Η Ζ— / ^F ^ O F Ο ^ Α, - / \ / ^Ν - ^Ζ 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3 , 4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000133 520.3 234 ^F \ / ^ OH Ν ί F η 7r- ~ / ^F v3 θ ^ Ν ^ ⁷ 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3 , 4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.00018 520.3

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235 ri QVu ’ θ O Ν 2-fluoro-1 - [(18, ZI) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4- B] indol-2-yl] -2methylpropan-1-one 0.024 482.3 236 Af JN - ⁷ CHAIR / (1I) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2fluoro-2-methylpropyl) - 1,3,4,9- tetrahydropyrido [3.4b] indole 0.000182 490.2 237 vf ^ X_7 N— 'GTU </ ^N gl ^h —O (18) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2fluoro-2-methylpropyl) - 1,3,4,9- tetrahydropyrido [3.4b] indole 0.046 490.2 238 O iz 1 AA \ oAJi J? [(1K, ZI) -1- [2,6-difluoro4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] - (1fluorocyclopropyl) methane one 0.000244 516.1 239 r— \ Ck ^ X Y N— 'XXXY -y O— ^ ^N - \ F (1R, 3R) -6-xnop-1- (2.6- difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.00156 538.1 240 / - \ / ^F JN— 'ΧΥνο У γ · Ά (1Р, ЗР) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -7fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1H- 0.000261 522.2

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pyrido [3,4-b] indole 241 Μ-Τ '' Ν i F ^Η У– / ^F <y ° Λ (1I, ZK) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.000381 522.3 242 / \ ^F ΧΧ / Τ Αγ Ο— F (1I, ZK) -1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl- 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.00542 522.3 243 4 / A M ^ Y \ x ^ == H Y - / ^ he N i F H l - ~ / ^F <7 ° h ^v t \ ^ ^f (1 - (((1R, 3R) -1- (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro -2Нpyrido [3,4-b] indol-2yl) methyl) cyclopropyl) methanol 0.002 514.2 244 .— \ / ~ ^F Ck Y ^N - 'YYHj ^H ^ -1 \ ^F V? O— \ F (1S, 3S) -6-xnop-1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3 , 4.9- tetra hydro-1 Npyrido [3,4-b] indole 0.063 538.1 245 to F n Y- / ^F v) ° y '^ ⁿ 0 ^ ^{/ F} (18.38) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.035 522.3

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246 0TU </ N 7 = / H _P / \ ^F v7 0 - F (1I, ZK) -1- [2,6-difluoro-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl2 - [(3-methyloxetane-3yl) methyl] -1,3,4,9 tetrahydropyrido [3,4b] indole 0.000358 536.1 247 F γ — 0 Af i TUCH / N / = < ^H c— / \ p \ ^N / - O— ' (1K, ZI) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -5fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.00434 522.2 248 / - \ / “ ^{F N} ~ ilY ^^ - j • 4 ° T ^and ^ u F (1K, ZI) -1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -7-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.000518 522.2 249 F> Lf N— ' L SPE.r 0— ^ N— \ F (1K, ZI) -1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -5-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl- 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.000511 522.2 250 __ __Z \ F \ ^ = / T N-Y N i H (1Р, ЗИ) -2- (2-fluoro-2methyl-propyl) -1 - [4- [2 [(3R) -3- (fluoromethyl) pyrrolidine -1 -yl] ethoxy] phenyl] -3methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.000275 482.2 251 I / - (/ ^F LTVA / ^H \ p?, —N 2-- 'O—' (18.38) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -5fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.021 522.2

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252 .— \ / ^-F У N— '> = < ^f- O 0— ^ N— \ F (18.38) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -7-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl- 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.05 522.3 253 F -_ζ in-F ^ \ ^ Y N ~ 'ίΓΜ / N> = < ^nr LZ O- ^ N- F (1S, 38) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -5-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl- 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.035 522.2 254 ° OtCy ^ 'T N' F H / r - / ^F V / ^ ⁿ O ^ \ F 2 - [(1R, 3R) -1- [2.6- d and fluoro p-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -M, M- dimethyl acetamide 0.04 515.2 255 > F УХ / У \ LZ ° ^H N 'F H АУ ^F ~ y ° Ά 3 - ((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro- 2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000010 520.2 256 Сч 1 ^s uz о i - F (1Р, ЗР) -2- (2-fluoro-2- methylpropyl) -1 - [4- [1 - (3fluoropropyl) azetidine-3yl] oxy-2-methylphenyl] 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.00169

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257 O (1I) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -2- (2fluoro-2-methylpropyl) - 3,3-dimethyl-4,9-dihydro-1 H-pyrido [3,4b] indole 0.00109 518.3 258 \ \ ^F ί TUCH / '^ X / - ^Ν / - o—' (8) -1- (2,6-difluoro-4- (2 (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2- (2-fluoro-2-methylpropyl) 3,3-dimethyl-2,3,4,9tetra hydro-1 Npyrido [3,4-b] indole 0.031 518.3 259 .— \ / F y \ 7 Ν— 'C X / UCh / ^f f% = <I .F H _p _ / \ r — Λ vX p 0 — P ^N ~' (1R, 3R) -1- [4- [1 - [(3,3difluorocyclobutyl) methyl] azetidin-3-yl] oxy2,6-difluoro-phenyl] -2- (2fluoro-2-methyl-propyl) 3 -methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000444 538.3 260 FH ^F ^ O ° A ^ CHUU (1Р, ЗР) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -8fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.000881 522.3 261 C TUCH / T Hr / \ F ^F ν / 0— ^ N— \ F (1Р, ЗР) -1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -8-fluoro2- (2-fluoro-2- methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.00028 522.3 262 <h - \ FF FH ^F V / SD (18.38) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -8fluoro-2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.026 522.3

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263 , —Ζ \ h11OH app ^F V? Ο- ^ Ν— F (S) -1 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3,4-b] indol-2yl) propan-2-ol 0.000359 488.3 264 οϋ Α 0— ^ Ν— \ F (R) -1 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3,4-b] indol-2yl) propan-2-ol 0.000969 488.3 265 τ <0 \ - l ^ΙΖ 1 Vz 7 ^1/1 (1R, 3R) -1- [4- [2- [3 (chloromethyl) azetidin-1 yl] ethoxy] -2,6-difluorophenyl] -2- (2-fluoro-2methyl-propyl) -3-methyl1, 3.4.9- tetrahydropyrido [3.4b] indole 0.0017 521.2 266 ___ __Λ \ F Ν i Q-c F (1R, 3R) -1- [3-xnop-4- [1 (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.01 503.2

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267 __ \ ^F N i H QO ^ A F (1 R, ЗR) -1- [3-φτop-4- [1- (3- phtoppropyl) azetidine-3yl] oxy-phenyl] -2- (2fluoro-2-methyl-propyl) - 3-methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.000744 486.3 268 / T ^H j_ / ^OH Ν i FH z— / ^F V7 o / F (2R) -3 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] propane-1,2diol 0.008 504.2 269 N 'F H / A / ^F ~ w O ^ A D * A. NA (1R, ЗR) -1- [2,6-difτop-4 [1 - [[(1S, 2R) -2- fluorocyclopropyl] methyl] azetidin-3-yl] oxyphenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.001 516.3 270 F 7n- / EXT / N GA ^Hf 'O \ -n / Xn _f (1R, ЗR) -1- (2,6-diφτop-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2 - ((8) 3-fluoro-2-methylpropyl) 3-methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000459 504.3

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271 OH 05 / ρ ^nr ~ O O- \ F (S) -3 - ((1 R, 3R) -1 - (2,6d and fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9- tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropane-1,2diol 0.005 518.3 272 [S = \ / - / F N i F H o ^ xx Λ ^ Ν-q ^ F (1R, ЗR) -1- [2,6-diφτop-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2- (3fluoro-2,2-dimethylpropyl) -3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000685 518.3 273 O IZ] / S ^V z ^, oLC ' ^v / X X / I (R) -2-φτop-3 - ((1R, ЗR) -1 (2-fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.00028 502.2 274 >. F z ^ \ / - ζ N i H As, ^f -q 0 ^ V _N ^ ^F (S) -2-φτop-3 - ((1R, ЗR) -1 (2-fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000253 524.1

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275 L * ^ζζ Ο (Ρ) -2-φτορ-3 - ((1Ρ, 3Ρ) -1 (4- (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000166 484.2 276 LL X Я Ό '^ ° ν' (8) -2-fluoro-3 - ((1H, 3H) -1 (4- (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000309 484.2 277 ν vaK ^ ⁿ ^ ^f Ν ί F Η / 55-Ζ Ο ^ χΛ F (1I, 3N) -1- [2,6-difluoro-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl2- (2,2,2-trifluoroethyl) - 1,3,4,9- tetrahydropyrido [3.4b] indole 0.000090 512.2 278 ΟΗ ο4. ^J H 0 ^ -Ν F (2R) -3 - [(1R, 3R) -1- [2,6 difluoro-4- [2- [3- (fluoromethyl) azetidine-1yl] ethoxy] phenyl] -3methyl-1,3,4,9tetrahydropyrido [3,4b] indol-2-yl] -2methylpropane-1,2-diol 0.00031 518.3

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279 O ZCZ O k / O ^ \ 1 G (1I, ZK) -2- (2-fluoro-2methylpropyl) -1 - (3fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-2,3,4,9tetra hydro-1 Npyrido [3,4-b] indole 0.00016 486.3 280 about IZ uk. GT k / SG] 1 Ή (18.3 ^ -2- (2-fluoro-2methylpropyl) -1 - (3fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.006 486.3 281 о tz / WV ^ * О Го θ '] Ή 1 Ή (1I, ZK) -1- (2,3-difluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000183 504.3 282 Ν 9 F Η L = Y Ac 0-Ο \ (1I, 3K) -1- (2,6-difluoro-4 ((1 - (((1S, 2S) -2- fluorocyclopropyl) methyl) azetidine-3yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.001 516.3 283 __ Г— / ° MAr ⁿ ^ Ν 'F Η ЛЮ Ό OO 1 - [(1R, 3R) -1- [2.6- difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] propan-2one 0.001 486.3 284 χχ ^F V? Ο— F 3 - [(1R, 3R) -1- [2.6- difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2,2 dimethylpropan-1-ol 0.000382 516.3

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285 O IZ J dGy (1I, ZK) -1- [2,6-difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2ethylsulfonyl-3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.002 522.1 286 c ^F \ L / ^0H N i F H 7 ^ / Άγ 3 - [(1R, 3R) -1- [2,6d and fluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4,9tetrahydropyrido [3 , 4b] indol-2-yl] -2,2 difluoro-propan-1 -ol 0.000097 eight 524.1 287 δ to z w / \\\ T [* ZT ^ O 3 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indol-2-yl] -2.2- dimethylpropan-1-ol 0.001 516.3 288 ... F OH __Λ h, Я __ / A 'n—' 05-4 / _F JH, ^f HN-A ^ (R) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [ 3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000052 eight 519.3 289 F OH iTV </ ^F ^ th FO hn-A) ⁿ ^ (S) -3 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidin-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000199 519.3

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290 ... F OH __Λ χχ ^p De and ΗΝ-φ ^1- ' (R) -2 ^ Top-3 - ((1R, 3R) -1 (2-fluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000497 501.3 291 F ОН _f - / Q> 7 ΗΝ — У ^м— (8) -2-fluoro-3 - ((1H, ZC) -1 (2-fluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.00013 501.3 292 ... F ОН __Λ h, 1 __ / T j HN — У ^{м- /} (H) -2-fluoro-3 - ((1I, ZTs) -1 (4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3methyl-1,3,4,9- tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.002 483.3 293 F OH ¹ b y T-Og (S) -2-φτop-3 - ((1R, ЗR) -1 (4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.00015 483.3 294 o _{/ Ζ} Ί nD ^, 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3 , 4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 520.3 295 __> o U to F H Ha / To/ 1 - ((1S, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3, 4-b] indol-2yl) propan-2-one 0.021 486.2

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296 rA ^F \ / ^F n g f n / === / ΗΝ = 3,5-d and fluoro p- N- (2- (3 (fluoromethyl) azetid in-1 yl) ethyl) -4 - ((1 R, 3R) -3methyl-2- (2,2,2trifluoroethyl) - 2,3,4,9 tetra hydro-1 Npyrido [3,4-b] indol-1 yl) aniline 0.0002 511.2 297 A Α Yy ° Y ^z y4j ^{z: CLJ} 'a (S) -3 - ((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3, 4.9 tetrahydro-2Hpyrido [3,4-b] indole-2πο) -Ν, Ν, 2trimethylpropanamide 0.018 543.3 298 ... / Wow (R) -3 - ((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3, 4.9 tetrahydro-2Hpyrido [3,4-b] indole-2πο) -Ν, Ν, 2trimethylpropanamide 0.005 543.3 299 o x 3 x ° / L W Ur- \ X G ^ζχ1ί_ o (S) -3 - ((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-L] indol-2yl) -2-methylpropanoic acid 0.003 516.2 300 y.§ ζΆ O (R) -3 - ((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-L] indol-2yl) -2-methylpropanoic acid 0.001 516.2 301 R / - \ / ^F > ^ 7 N— 'STU-C / A O— ^ N— \ F (F, 38) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -2- (2-fluoro-2-methylpropyl) -3 (fluoromethyl) -2,3,4,9 tetra hydro-1 Npyrido [3,4-b] indole 0.000088 1 522.2

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302 y - / ^F J / ^0H N 'F H TA - / o-Y \ ^ - / ^F 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3- yl) oxy) phenyl) -3-methyl3,4-dihydro-1H- pyrido [3,4-b] indole- 2 (9H) -yl) -2.2- difluoropropane-1 -ol 0.00012 524.1 303 QaX ~ ^f Ν г F н (7 (1R, 3R) -2- (2,2- difluoroethyl) -1 - [2,6 difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indole 0.000048 2 494.2 304 V F N b F H / = / M- (3,5-difluoro-4 ((1R, 3R) -3-MeTnn-2 (2,2,2-trifluoroethyl) 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.000224 511.2 305 Q £ <> - 'N ^X ί F H 7r-i ^F ^ O Ο-, ^ ~~ ^N \ d ~ y F (1R, 3R) -2- (2,2- difluoroethyl) -1 - [2,6 difluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000216 494.2 306 »F ₄ F \ —F _ ['N— CdAa h _f _q ⁴ F (1S, 3R) -2- (2,2difluoroethyl) -1 - [2,6difluoro-4- [2- [3 (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3methyl-1,3,4, 9tetrahydropyrido [3.4b] indole 0.003 494.2

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307 A \ F X / “i N— z. F ^hf ^ _F 'Ά (1 R, 3R) -1- [4- [1 - (3,3difluorocyclobutyl) azet idin-3-yl] oxy-2,6difluoro-phenyl] -2- (2fluoro-2-methylpropyl) 3-methyl -1,3,4,9tetrahydropyrido [3,4b] indole 0.000193 534.1 308 O IZ I -П (1Р, ЗР) -1- [2,6-difluoro-4 [1 - [(E) -3- fluoroallyl] azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000127 502.3 309 adT -CH 0— F (Sh, ZR) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -3-methyl2- (2- (methylsulfonyl) propyl) -2,3,4,9-tetrahydro1 H-pyrido [3,4-b] indole 0.006 550.3 310 ΓΆ / \ о N i О H О ^HN 4> ^ F 1 - (3-fluoropropyl) -M- [4 [(1Р, ЗР) -3-methyl-2methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole-1- yl] phenyl] azetidine-3amine 0.000708 471.2 311 GA / lo OP_ / ^N 'sA X ^ x --- ζ C n IA d ~ h HN ^ A F M- [3,5-difluoro-4 [(1P, 3R) -3-methyl-2methylsulfonyl1,3,4,9tetrahydropyrido [3,4b] indol-1-yl] phenyl] -1 (3fluoropropyl) azetidine-3amine 0.000248 507.2

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312 Ζ * o \ Jr4 ^K ~ ^ N i O N O (1R, 3R) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-2-mpu1sulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.002 484.1 313 / ^ \ / n o Uk'iv N i 0 ^{n n} Z ^ O ° A (1R, 3R) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -M, 3-dimethyl-1,3,4,9tetrahydropyrido [3,4b] indole-2-sulfonamide 0.001 487.4 314 CTX / N'SX £ 14 o N O HN ^ zA F 1 - (3-fluoropropyl) -M- [4 [(1S, 3R) -3-MeTnn-2methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole-1- yl] phenyl] azetidine-3amine 0.009 471.3 315 χΑ _ / \ θ CZX / ^N 's Ла ^F <5 ην- ^ ζΑ F M- [3,5-difluoro-4 [(1S, 3R) -3-MeTnn-2methylsulfonyl1,3,4,9tetrahydropyrido [3,4b] indol-1-yl] phenyl] -1 (3fluoropropyl) azetidine-3amine 0.002 507.1 316 О ιζ 1. А / A iii \ z z 3 - [(1R, 3R) -1- [2.6- difluoro-4- [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3.4b] indol-2-yl] -2.2- dimethylpropanenitrile 0.000072 4 511.2

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317 N i FH / == - / ^F ~ wx F o ^ / A / == / \ .N-Y Λ F (1 R, 3R) -1- [4- [1 - (3,3difluoroallyl) azetidin3-yl] oxy-2,6-difluorophenyl] -2- (2-fluoro-2methylpropyl) -3-methyl1,3 , 4.9 tetrahydropyrido [3.4b] indole 0.000181 520.1 318 OH </ H n ~ ^ A ^ f x = ^ H Y - / F th i-7 '0--. (S) -2 - (((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3 , 4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) methyl) -3,3,3trifluoropropane-1 -ol 0.001 556.2 319 OH N i F H z— / ^F V / ° Ί y ^F (R) -2 - (((1R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-1,3 , 4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) methyl) -3,3,3trifluoropropane-1 -ol 0.00043 556.2 320 y - \ L / ^0H νχ ~ ν ^ ^Ν ^^ N i FH / == / py HN ^. 3 - ((1R, 3R) -1- (2,6d and fluoro p-4 - ((2- (3 (fluoromethyl) azetid in-1 yl) ethyl) amino) phenyl) 3-methyl-1,3, 4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.00027 519.2 321 > \ ^F U N— OZch / ^N ίΛ ^H FO HN— \ / H_ X— N / - \ X / F 3,5-difluoro-M- [2- [3 (fluoromethyl) azetid in-1 yl] ethyl] -4 - [(1Р, ЗР) -2- (2fluoro-2-methyl-propyl) 3-methyl-1 , 3,4,9tetrahydropyrido [3,4b] indol-1-yl] aniline 0.000222 503.2

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322 > \ ^F __Γ ν— '0 HN- \ 3-φτορ-Ν- [2- [3 (fluoromethyl) azetid in-1 yl] ethyl] -4 - [(1 R, 3R) -2- (2fluoro-2-methyl-propyl) 3-methyl-1, 3,4,9tetrahydropyrido [3,4b] indol-1-yl] aniline 0.000025 3 485.2 323 > \ ^F JN— OUch Q HN - '\ / K—' X — N / \ F Ν- [2- [3- (fluoromethyl) azetid in-1 yl] ethyl] -4 - [(1 R, 3R) -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole -1-yl] aniline 0.000215 467.2 324 / GA Ζλ o Uk / ^N 'S ^ N io H О (1R, 3R) -2- ethylsulfonyl-1 - [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.000198 486.2 325 HA / y about LH / ^N A N i 0 H O ΗΝ ^. F N- [2- [3- (fluoromethyl) azetid in-1 yl] ethyl] -4 - [(1 R, 3R) -3methyl-2methylsulfonyl1,3,4,9- tetrahydropyrido [3,4b] indol-1-yl] aniline 0.001 471.1 326 About ιζ ADD 3,5-difluoro-N- [2- [3 (fluoromethyl) azetid yn-1 yl] ethyl] -4 - [(1 R, 3R) -3methyl-2methylsulfonyl1,3,4,9- tetrahydropyrido [3,4b] indol-1-yl] aniline 0.000305 507.2

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327 \ Gon 'U ~ L / ⁿ ' ⁷ N i H HN ^ 2-fluoro-3 - ((1I, ZK) -1- (2fluoro-4 - ((2- (3- (fluoromethyl) azetid in-1 yl) ethyl) amino) phenyl) 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000347 501.2 328 \ ΓθΗ N i n / a HN ^ 2-fluoro-3 - ((1I, ZK) -1- (4 ((2- (3- (fluoromethyl) azetid in-1 yl) ethyl) amino) phenyl) 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropan-1-ol 0.000167 483.3 329 λΆ / \ ° Qvf < About n ^ f (1R, 3R) -1- [4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -M, M, 3trimethyl-1,3,4,9tetrahydropyrido [3,4b] indole-2-sulfonamide 0.001 523.1 330 / ^ \ / \ 0 (P / DX N =. O HA O \ F O ^ A, - / \ / ⁿ ~ A (1R, 3R) -1- [4- [1- (3- fluoropropyl) azetidine-3yl] oxyphenyl] -3-methyl2-methylsulfonyl- 1,3,4,9- tetrahydropyrido [3.4b] indole 0.002 472.2 331 A \ F / / ”5 MX ^N ΐ> 7 H / == <F - / \ νγ θ '// ¹ /' - // \ ΖΌΗ 3- [3- [3,5-difluoro-4- [(1 R, ЗR) -2- (2-φτop-2methyl-propyl) -3-methyl1,3,4,9- tetrahydropyrido [3,4- B] indole-1- yl] phenoxy] azetidin-1 yl] cyclobutanol 0.000236 514.2

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332 O IZ J OLA / i ▼ (1 R, 3R) -1 - (4- (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -2 - ((8) isopropylsulfinyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.008 484.2 333 « ^{; Ο} v * ГУ ν 'О (1 R, 3R) -1 - (4- (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -2 - ((P) isopropylsulfinyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.048 484.2 334 f \ FX / “ί N-- ~ Z ~ NA f ^H _F j4 ^{4γ 0} ChHm //./ \ \ X '' / OH (18,3h) -3- (3- (3,5-difluoro4 - ((1Р, ЗР) -2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1 Npyrido [ 3,4-b] indol-1 yl) phenoxy) azetidin-1 yl) cyclobutanol 0.00013 514.2 335 z ^^ z ZI θ (1Р, ЗР) -1- [2,6-difluoro-4- [1- (5- fluoropentyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) - 3-methyl-1,3,4,9- tetrahydropyrido [3.4b] indole 0.000442 532.3 336 αΓζ, Cho-g ^ F (1Р, ЗР) -1- [3,5-difluoro-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000263 504.3

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337 О IZ D. XT L ζ (1I, ZK) -1- [2,6-difluoro-4 [1- (4- fluorobutyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.000387 518.3 338 ^Η O ~ ^F F ° -Q- \ F (1K, ZI) -1- [3,5-difluoro-4 [1- (5- fluoropentyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.001 532.2 339 __> \ F γυΟ ^ ^η Άα Q — γ F (1K, ZI) -1- [2,5-difluoro-4 [1- (3- fluoropropyl) azetidine-3yl] oxy-phenyl] -2- (2-fluoro-2-methylpropyl) 3-methyl-1,3,4,9tetrahydropyrido [3,4b] indole 0.001 504.2 340 y - ^F \ E / ^0H Ν Ь F H 7AV ^f ~ a HN ^ / X F 3 - [(1R, 3R) -1- [2.6- difluoro-4 - [[1- (3- fluoropropyl) azetidine-3yl] amino] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2,2 difluoro-propan-1-ol 0.000203 523.2 341 \ F ΟΑ5 ^ν Ά ⁿ Q ° Ά ^ν ~ \ F 3 - [(1R, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethylamino] phenyl] -3methyl-1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2,2 difluoro-propan-1-ol 0.000348 545.2

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342 СЮ Ν 9 n / = l Ν 1. °% _Ν ^ ^/ (1S, ЗR) -2- (2-φτop-2methylpropyl) -1 - (5 - ((1 (3- fluoropropyl) azetidine-3yl) oxy) pyrazin-2-yl) -3methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000619 470.3 343 __ E z Ν ’- F Η / = / J Ay) \ Γ ~ Χ o-Lj (1K, ZI) -1- (2,6-difluoro-4 ((1 - (3-fluoropropyl) -Zmethylazetidin-3yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3- methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.0002 518.3 344 ϊ \ F οΌ ^ν ~ γ ^xX "e / ^Η Ο ° Ο ^ν ~ \ F 2 - [(1 R, ЗR) -2- (2-φτop-2methyl-propyl) -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-1-yl] -5- [1 - (3fluoropropyl) azetidine-3yl] oxy-benzonitrile 0.000347 493.3 345 __ / F. / Ν 9 ^Η Ο Hu ⁰ 4 - ((1 R, ЗR) -2- (2-φτop-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indol-1 yl) -1 - ( 3- (3- (fluoromethyl) azetid in-1 yl) propyl) pyridin2 (1H) -one 0.0123 483.3 346 __> \ F рхО ··· 'Ν ^Ζ '> __ ^Η Ο УхХ-л F [4 - [(1R, ЗR) -2- (2-φτop-2methyl-propyl) -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-1 -yl] phenyl] - [1 - (3- fluoropropyl) azetidine-3yl] methanone 0.000399 480.2

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347 SCRL ° _F nh ₂ n / g - / (R) -3 - ((1R, 3R) -1- (2,6 difluoro-4- (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropanamide 0.014 515.3 348 ΟΧ / Ά ^οη Ν to F Ο 'ό Ο- ~ Ά F (R) -3 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropanoic acid 0.001 516.2 349 Ο τζ] ίγ sA γ ° Κ / ο (S) -3 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-methylpropanoic acid 0.00203 515.2 350 τ \ ο / - ^ ° I ^{ζτ w} Ο 3- [1- [2,6-difluoro-4- [1- (3fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2methylpropanoic acid 0.014 516.2 351 n γγ ° f-0 Ο-γΑ ___. F > ^ m ----- 3- [1- [2,6-difluoro-4- [1- (3fluoropropyl) azetidine-3yl] oxy-phenyl] -3-methyl1,3,4,9- tetrahydropyrido [3,4b] indol-2-yl] -2methylpropanoic acid 0.033 516.2 352 F y - Λ> r, r he Ν i F Η Aaa / ΗΝ ^ (R) -3 - ((1R, 3R) -1- (2,6d and fluoro-4 - ((2- (3- (fluoromethyl) azetidin-1yl) ethyl) amino) phenyl) 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000685 519.3

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353 ... F / —ς <? He N i F H 4V ^F T HN ^ ^ ЧУ ^^ (8) -3 - ((1 R, 3R) -1- (2.6d and φτορ-4 - ((2- (3- (fluoromethyl) azetidin-1yl) ethyl) amino) phenyl) 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000033 519.3 354 DH / ~ -ξ \ / ° F ^OH H _ / ^F ^ O NN ~ -D \ y - / X NV (2R) -3 - [(1R, 3R) -1- [2.6- difluoro-4 - [[1- (3- fluoropropyl) azetidine-3yl] amino] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3,4- B] indol-2-yl] -2methylpropanoic acid 0.001 514.2 355 Y — D ^F. F ^f to F n X = / Ό (18.38) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-3-methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Npyrido [3,4-L] indole 0.0292 530.2 356 D. ~ S ^F. F \ ~ Lx Ν;. F Η XV O Tv ~ OV (1Р, ЗР) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-3-methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Npyrido [3,4-L] indole 0.000509 530.2 357 ^ IZ ζ τι (1S, 38) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro3-methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Hpyrido [3,4-b] indole 0.0261 529.5

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358 va ~ k3 ^ ^<f N i FH 5 = / Ο ^ ζΛ F (1I, ZK) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro3-methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Hpyrido [3,4-b] indole 0.000417 529.5 359 F ₄ > F - \ ^f \ Lz ° ^h vOX '·' N i F H 5 = / ^F ~ w ° ~ Ά 3 - ((1 R, 3R) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-3-methyl-1,3 , 4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2,2-difluoropropan-1 ol 0.000552 541.5 360 ex 1¾? ^s 3 - ((1S, 3S) -1- (2,6d and fluoro p-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -6fluoro-3-methyl-1,3, 4,9 tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2,2-difluoropropan-1 ol 0.009 541.5 361 / ^ ^F \ / Л ^N ~ ^ If N i F ^F H - / ^Ρ -θ ° Ά ^ ^Ν ζΧ ^ / ^Ρ (1R, ЗR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) azetid yn-1 yl] ethoxy] phenyl] -3- methyl-2- (3,3,3- trifluoropropyl) -1,3,4,9 tetrahydropyrido [3,4- B] indole 0.003 525.5 362 rw ^ / ^ ^F \ / Л ^N - ^F F ^F h JA / ^F v3 ° A (1S, ЗR) -1- [2,6-difτop-4 [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-2- (3,3,3- trifluoropropyl) -1,3,4,9tetrahydropyrido [3,4b] indole more than 0.1 526.2

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363 ^F \> _P F / - ^{r F} \ L / ^0H N i F H 7 - / ^F V / o-Y \, - / ^F 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2,2-difluoropropan-1 ol 0.000166 542.1 364 F. AF C N-Y ^/ N '^ VF H J / ^F o- ~ Y \. -Z ^F χ ^ Ν-Y 3 - ((1 S, 3S) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro 3-methyl-1,3,4,9tetrahydro-2H - pyrido [3,4-b] indol-2yl) -2,2-difluoropropane-1 - ol 0.009 542.1 365 F.> F \ === \ / - ζ Αρη N i F H А / ^f Q ΗΝ - ^ / Λ F 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2,2-difluoropropan-1 ol 0.000233 563.2 366 RF UA — ZA ^F \ // ^OH n'v f H Ж ^f ~ A HN-уL X ^ N - ^^ F 3 - ((1 S, 3S) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2,2-difluoropropan-1 ol 0.004 563.2 367 F = \ / - Z \ 0 f ^oh η 7- / ' ^F HN-UL X- ~ Z \ / ^N - ^Z (2S) -3 - [(1R, 3R) -1- [2.6- difluoro-4 - [[1- (3- fluoropropyl) azetidine-3yl] amino] phenyl] -3methyl-1,3,4,9- tetrahydropyrido [3,4- B] indol-2-yl] -2methylpropanoic acid 0.008 515.3

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368 ^F V ^ r - / ^F y ^ OH (aX ⁿ ^ N b TH ', F 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000546 538.3 369 ^F \ F V ^0H yA ^- C / ^N '''N ^ \ F H / = / ^f O \ / V> ^F 3 - ((1 S, 3S) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro- 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.024 538.3 370 5 "ChuO ' ⁰ ζΥ Ο 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000203 538.3 371 ^F \ '''F V / OH C / Ν - /> Ν ^ \, F Η / = / ^f O F 3 - ((1 S, 3S) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) oxy) phenyl) -6-fluoro- 3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.009 538.3 372 ^F Vx ν Ν i F Η / = / ^Ρ Ό F ΗΝ - ^ / Λ ---- M- (3,5-difluoro-4 ((1R, 3R) -6 ^ Top-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Hpyrido [3,4-b] indole -1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.000198 529.2 373 ^F y ~ xX V yA "to FH K- / ^F ~ w F HN-yL y - l ^ dm- / M- (3,5-difluoro-4 ((18.38) -6-fluoro-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Hpyrido [3,4-b] indole -1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.018 529.2

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374 ΛΓ ⁿ ₽-O XZ \ .NJ o 3 - ((1 R, 3R) -1 - (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -3methyl-1,3,4,9tetrahydro-2Npyrido [ 3,4-b] indol-2- yl) -2.2- dimethylpropanoic acid 0.00155 530.3 375 OH __ / ~ Y -. F \ ν-ΑΆ AA F ^F H / = / ^F ~ O \ F (R) -2 - (((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro-2Hpyrido [3,4-b] indol-2yl) methyl) -3,3,3trifluoropropan-1-ol 0.000764 556.2 376 OH __.— Z <F </ ¾ nJ% 'nA f ^f Ό \, F (S) -2 - (((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) oxy) phenyl) -3-methyl1,3,4,9-tetrahydro -2Нpyrido [3,4-b] indol-2yl) methyl) -3,3,3trifluoropropane-1 -ol 0.001 556.2 377 __ / E / N г ^H ζΛ VN N% \. F (Ш, ЗР) -2- (2-fluoro-2methylpropyl) -1 - (2 - ((1 (3- fluoropropyl) azetidine-3yl) oxy) pyrimidine-5yl) -3-methyl-2,3,4,9tetra hydro-1 Npyrido [3,4-b] indole 0.0005 470.3 378 p = \ / --f V 0 \ Fl, Ν ^ Ί F ° ^{H F} Λζ 3 - [(1S, 3R) -1- [2.6- d and fluoro-4- [2- [3- (fluoromethyl) azetid in-1 yl] ethoxy] phenyl] -3- methyl-1,3,4,9- tetrahydropyrido [3.4b] indol-2-yl] -2.2- dimethylpropanoic acid 0.028 530.3

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379 N g F n l - / f ρΝ F-U (1 / ?, 3 /?) - 1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2- (2,2difluoropropyl) -3-methyl2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indole 0.00028 508.2 380 b \ F ^4Ν "^ \ 4-ν Η \ / HO F (S) - (4 - ((1 R, 3R) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1Hpyrido [3,4-b] indol-1 yl) phenyl) (1- (3- fluoropropyl) azetidine-3yl) methanol 0.000188 482.3 381 > \ ^F > Λ HO F (R) - (4 - ((1R, 3R) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9tetrahydro-1H- pyrido [3,4-b] indol-1 yl) phenyl) (1- (3- fluoropropyl) azetidine-3yl) methanol 0.000605 482.3 382 AR Ν— ⁷ OH L TUCH J ^H f4 7 г- / \ / x ^r \ - / ^ N ^ F N ^ H 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -3methyl-1,3,4,9tetrahydro-2H- pyrido [3,4-b] indol-2yl) -2,2- dimethylpropanoic acid 0.00101 528.2 383 / ^F \ __ / \ - F F \ P N- ⁷ VTV4 / n GA ^h fO 0 r ^N F- ⁷ (1 / ?, 3 /?) - 1- (2,6-difluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -2- (2,2 difluoroethyl) -6-fluoro-3- methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.00028 512.2

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384 _г Ε '/ ^F К ^N ^ ¥ TV </ ^f aa ya 0 < ^N FZ (15.35) -1- (2,6-difluoro-4 (2- (3- (fluoromethyl) azetidine-1yl) ethoxy) phenyl) -2- (2,2difluoroethyl) -6-fluoro-3- methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.029 512.2 385 ^F \ __ x / ^F \ N g F H z— / / F (1 / ?, 3 /?) - 1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -2- (2,2 difluoroethyl) -6-fluoro-3- methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000074 512.2 386 _d F. ^? / ^F К N— / YYV </ ^F A ^ ⁿ YA 0 ύ F (15,35) -1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -2- (2,2difluoroethyl) -6-fluoro-3methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.023 512.2

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387 R / 'th ^F ~ O NH F L / - (3,5-difluoro-4 ((1 / ?, 3 /?) - 6-fluoro-3methyl-2 (methylsulfonyl) 2,3,4,9-tetrahydro-1 Npyrido [3,4-b ] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.000149 525.2 388 F λ 0 ^Χ ή '\ O _f h V / ^F vj NH F / \ / - (3,5-difluoro-4 ((1δ, 3δ) -6-φτορ-3methyl-2- (methylsulfonyl) - 2,3,4,9-tetrahydro-1 Npyrido [3,4-b] indol-1 yl) phenyl) -1- (3- fluoropropyl) azetidine-3amine 0.019 525.2 389 , ^F \ _ / \ —F F _{x /} x J ^{Ν- 0} YTh j ^nr Ч4 NH i / F / V- (4 - ((1R, 3R) -2- (2,2 difluoroethyl) -6-fluoro-3methyl-2,3,4,9tetrahydro-1 / 7pyrido [3,4-b] indol-1 yl) -3,5-difluorophenyl) 1- (3- fluoropropyl) azetidine-3amine 0.000056 511.2

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390 ^{'F \' \ -F F. N} - Y YtVa / ^nr SE NH F / V- (4 - ((1S, 3S) -2- (2,2 difluoroethyl) -6-fluoro-3methyl-2,3,4,9tetrahydro-1 / 7pyrido [3,4-b] indol-1 yl) -3,5-difluorophenyl) 1- (3- fluoropropyl) azetidine-3amine 0.0097 511.2 391 __Z F AO ^- M - / n g f H l - / hh o F ^ c '' (1 / ?, 3 /?) - 1- (2,6-difluoro-4- ^ -cis- (3- (fluoromethyl) cyclobutyl) azetidine-3- yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3- methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.00015 530.3 392 __Z F Co CL n g f H z— / ^F X (1 / ?, 3 /?) - 1- (2,6-difluoro-4- ((1 -trans- (3- (fluoromethyl) cyclobutyl) azetidine-3- yl) oxy) phenyl) -2- (2- fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetrahydro-1 Npyrido [3,4-b] indole 0.00044 530.3 393 wow About zY O (1I, ZK) -1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) oxy) phenyl) -2 - ((1 fluorocyclobutyl) methyl) - 3-methyl-2,3,4,9- tetrahydro-1 H- pyrido [3,4-b] indole 0.000142 516.3

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394 _B OH 7 N'T ¹ n w. ^Hf --O о r ^N F— ' (S) -3 - ((1 R, 3/7) -1- (2.6d and fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9 / - /) - yl) -2-fluoropropan1-ol 0.00023 506.2 395 _F ^OH J N- ^ ^/ (Y> 4 j ^Hf 'O o r ^N F—' (/ 7) -3 - ((1 R, 3/7) -1 - (2,6d and fluoro-4- (2- (3 (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3 , 4-dihydro-1 Npyrido [3,4-b] indol2 (9 / - /) - yl) -2-fluoropropan1-ol 0.000053 506.2 396 ^F y / N g F H z— / ^Ρ Ύ3 NH A F L / - (3,5-difluoro-4 ((1 / 7,3 / 7) -7-fluoro-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1 / 7pyrido [ 3,4-b] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.00011 529.2 397 --Λ ^F \ / ^F _f 4J £ ⁿ ^ ^f H Y-Y ^F VJ NH u F L / - (3,5-difluoro-4 ((1δ, 3δ) -7-φτορ-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1 / 7pyrido [3,4- B] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.033 529.2

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398 F r-X \ Ζ ^ οη Ν 7 F Η ΖΎ ΝΗ ίϊ F (S) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -5fluoro-3-methyl-3,4dihydro-1 / 7-pyrido [3,4 £>] indol-2 (9 / - /) - yl) -2fluoro-2-methylpropan-1 ol 0.00011 537.3 399 F _χ / m-X / - <'Υ ^ ΟΗ \ Χ ^ ^Ν ' ~ / ^F F n Λ ~ Ζ ^F v3 NH F (R) -3 - ((1S, 3S) -1- (2.6- difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -5fluoro-3-methyl-3,4-dihydro-1/7-pyrido [3.4 £>] indole-2 (9 / - / ) -yl) -2fluoro-2-methylpropan-1 ol 0.0011 537.3 400 F dX / - (\ ^ OH N 7 F h ZV ^F T0 NH 7 F (R) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -5fluoro-3-methyl-3,4dihydro-1 / 7-pyrido [3,4b] indol-2 (9 / - /) - yl) -2fluoro-2-methylpropan-1 ol 0.000023 537.3

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401 F _h / gA / - \ AOH N ^Z \ F HW ^F <7 NH nA F (S) -3 - ((1S, 3S) -1- (2.6- difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -5fluoro-3-methyl-3,4 dihydro-1 / - / - pyrido [3,4b] indole-2 (9 / - /) -yl) -2fluoro-2-methylpropan-1 ol 0.0024 537.3 402 F \ / A N - / A f ^oh J F ^ Z ^ (R) -3 - ((1R, 3R) -1- (2,6d and fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9H) -yl) -2- (fluoromethyl) propane-1 -ol 0.000032 520.3 403 F ^ ZA f ^oh h ^F ^ 3 f, Z ^N (S) -3 - ((1R, 3 /?) - 1- (2.6d and fluoro-4- (2- (3- (fluoromethyl) azetid in-1 yl) ethoxy) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3,4-b] indol2 (9H) -yl) -2- (fluoromethyl) propane-1 -ol 0.00013 520.3 404 Qi / Z N L F n / = / \ F Z ^ N-V ^ M- (3,5-difluoro-4- ((1I, 3R) -2- (2-fluoro-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indol-1 yl) phenyl) -1- (3fluoropropyl ) -Mmethylazetidine-3-amine 0.000745 517.3

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405 ZX ^F \ N g F NH F (R) - / V- (4- (2- (2,2difluoroethyl) -3,3dimethyl-2,3,4,9tetra hydro-1 / 7pyrido [3,4-b] indol-1 yl) -3, 5-difluorophenyl) 1- (3- fluoropropyl) azetidine-3amine 0.00023 517.3 406 ^F \ N ^z> \ F HA ^F V / NH F (S) - / V- (4- (2- (2,2difluoroethyl) -3,3dimethyl-2,3,4,9tetrahydro-1 / 7pyrido [3,4-b] indol-1 yl) -3.5 -difluorophenyl) 1- (3- fluoropropyl) azetidine-3amine 0.0018 507.3 407 F rX / - / ^f v ^f A ₌ aa_ / ^{n F} NEF H / - / ^Ρ Ύ3 NH F L / - (3,5-difluoro-4 ((1 / ?, 3 /?) - 5-fluoro-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1 / 7pyrido [ 3,4-b] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.00012 529.2

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408 F rX / Т ^F V ^F \ PP ^N X ' ^F n'v f ^F NH nT F L / - (3,5-difluoro-4 ((1δ, 3δ) -5-φτορ-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1 / 7pyrido [3,4- B] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.0063 529.2 409 GL / - / ^F \ / ^{f F} f FH Л- / ^F vJ NH F L / - (3,5-difluoro-4 ((1δ, 3δ) -8-φτορ-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1 / 7pyrido [3,4- B] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.026 529.2 410 / —Y ^F V ^F PXX ^ ⁿ --T ' ^f / Ή F FH 2 - / ^F ^ 0 NH F L / - (3,5-difluoro-4 ((1 / ?, 3 /?) - 8-fluoro-3methyl-2- (2,2,2trifluoroethyl) -2,3,4,9tetrahydro-1Hpyrido [3, 4-b] indol-1 yl) phenyl) -1- (3fluoropropyl) azetidine-3amine 0.00012 529.2 411 , __ Λ 1—0 TY and k / ^f O N ^^ F (1I, ZK) -1- (2,6-difluoro-4 (1- (3- fluoropropyl) azetidine-3yloxy) phenyl) -2 - ((3fluoroxetane-3yl) methyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.00007 518.3

- 80 038835

412 / HA / --f \ Z ^{OH f} aJa ⁿ ^ ^f F H ЛА ^F NH N- ^ F (S) -3 - ((1S, 3S) -1- (2.6- difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4 dihydro-1 / - / - pyrido [3,4b] indole-2 (9 / - /) -yl) -2fluoro-2-methylpropan-1 ol 0.0013 537.3 413 l-Xk Z ~ \ \ X ^{OH f} a / a ⁿ ^ ^f n g f Η z— / NH s F (R) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4dihydro-1 / 7-pyrido [3,4b] indol-2 (9 / - /) - yl) -2fluoro-2-methylpropan-1 ol 0.000018 537.3 414 d - ^ / ° ^H N; FH y - / NH s F (S) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4dihydro-1 / 7-pyrido [3,4b] indol-2 (9 / - /) - yl) -2fluoro-2-methylpropan-1 ol 0.00012 537.3 415 rA - / - \ A-oh f / JIa ^F N ^ YF Η A ^F A? NH z F (R) -3 - ((1S, 3S) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4dihydro-1 / 7-pyrido [3,4b] indol-2 (9 / - /) - yl) -2fluoro-2-methylpropan-1 ol 0.0095 537.3

- 81 038835

416 F aC ζ ~ ά t-oh vCL / '' ^{7 F} N 1 FH l - / NH X F 3 - ((1 R, 3R) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -5fluoro-3-methyl-3,4-dihydro-1/7-pyrido [3,4 £>] indol-2 (9 / - /) - yl) -2,2 difluoropropane-1 -ol 0.000024 541.2 417 F aC / -d 'Voh F H A- / ^F L7 NH X F 3 - ((1 S, 3S) -1 - (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidine-3yl) amino) phenyl) -5fluoro-3-methyl-3,4-dihydro-1/7-pyrido [3,4 £>] indol-2 (9 / - /) - yl) -2,2 difluoropropane-1 -ol 0.0033 541.2 418 OH ίΓγ ^ Λ < ^N ---- / ^ N N 1 F Η / - / NH F (R) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -3methyl-3,4-dihydro-1 Npyrido [3 , 4-b] indol2 (9H) -yl) -2-fluoro-2- (hydroxymethyl) propane ytrile 0.000033 530.3

- 82 038835

419 ОН zwa ^f £ _ c n-у N i FH Ж / NH F (S) -3 - ((1R, 3R) -1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -3methyl-3,4-dihydro-1Npyrido [3 , 4-b] indol2 (9 / - /) - yl) -2-fluoro-2- (hydroxymethyl) propane ytrile 0.000026 530.3 420 / t-X / ^ \ ^F \ / - OH Ν ί F ^N DCH NH D F (/?) - 3- (1- (2,6-difluoro-4- ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3,3dimethyl-3,4-dihydro1 / 7-pyrido [3,4-b] indol2 (9H) -yl) -2,2difluoropropan-1-ol 0.000117 537.3 421 / gA. / - m ^F \ Z ^ ^0H ^ 4XXz ^N '~ A' ^F N ' ^Z \ F n Ж / ^F V? NH D F (5) -3- (1- (2,6-difluoro-4 ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -3,3dimethyl-3,4-dihydro1 / 7-pyrido [3,4-b] indol2 (9H) -yl) -2,2difluoropropan-1-ol 0.000146 537.3

- 83 038835

422 / = \ / ~ ά \ / - he F FH Л- / ^F vJ NH ЛЭ F 3 - ((1 S, 3S) -1- (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -8fluoro-3-methyl-3,4 dihydro-1 / - / - pyrido [3,4 £>] indol-2 (9 / - /) - yl) -2,2 difluoropropane-1 -ol 0.0371 541.2 423 ✓ F _ F = \ / Ά \ Z ~ -oh Chpd- ^{7 F} / Ή s F FH z —- / ^F 30 NH X F 3 - ((1 / ?, 3 /?) - 1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -8fluoro-3-methyl-3,4 dihydro-1/7-pyrido [3,4 £>] indol-2 (9 / - /) - yl) -2,2 difluoropropane- 1-ol 0.000089 541.2 424 X ^ rSU ^ NE ^{F F Hp} 4 ° Άν-, F (1I, ZI) -1- (2,6-difluoro-4 (1- (3- fluoropropyl) azetidine-3yloxy) phenyl) -6,8- e and fluoro p-2- (2-fluoro p-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.0005 540.3 425 / F _ / ГЭх / - (\ Z ^ OH kJ4J ⁿ ^ ^f N г FH / - / NH У F 3 - ((1 / ?, 3 /?) - 1- (2,6difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4- dihydro-1/7-pyrido [3.4 £>] indol-2 (9 / - /) - yl) -2,2 difluoropropan-1 -ol 0.000021 541.3

- 84 038835

426 / GKhch / —l ' ^f \ Zoh N ^Z \ F n L- / ^F V7 NH U F 3 - ((1 S, 3S) -1- (2,6 difluoro-4 - ((1- (3fluoropropyl) azetidin-3yl) amino) phenyl) -7fluoro-3-methyl-3,4-dihydro-1/7-pyrido [3,4b] indol-2 (9/7) -yl) -2,2 difluoropropane-1 -ol 0.00638 541.2 427 ^F \ / N r F H / - / y (1R, 3/7) -1- (2,6-difluoro-4 ((1 - ((1- (fluoromethyl) cyclopropyl) methyl) azetidine-3yl) oxy) phenyl) -2- (2fluoro-2-methylpropyl) -3methyl-2,3,4,9- tetra hydro-1 Npyrido [3,4-b] indole 0.000278 530.3 428 R> / - ^ \ ^F F-Yy / Y / NTx and K / ^P y ^p ° An3 (1I, ZR) -1- (2,6-difluoro-4 (1- (3- fluoropropyl) azetidine-3yloxy) phenyl) -6,7- e and fluoro p-2- (2-fluoro p-2methylpropyl) -3-methyl2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indole 0.0007 540.3 429 and z— (1 / ?, 3 /?) - 2- (2-fluoro-2methylpropyl) -1 - (1 - ((1 (3- fluoropropyl) azetidine-3yl) methyl) -1 / 7-pyrazol-4yl) -3-methyl-2,3,4,9tetra hydro-1 Npyrido [3,4-b] indole 0.00099 456.3 430 __ / F О ~ С / П ^р N - F H z— / ^F Vv NH F N- [4 - [(1R, 3R) -2- (2,2difluoroethyl) -3-methyl1,3,4,9tetrahydropyrido [3,4b] indol-1-yl] -3,5difluoro-phenyl] -1- (3fluoropropyl) azetidine-3amine 0.000065 493.2

- 85 038835

Administration of compounds of formula I

The compounds of this invention can be administered by any route appropriate to the condition to be treated. Suitable routes include oral, parenteral (including subcutaneous, intramuscular, intravenous, intraarterial, intradermal, intrathecal, and epidural), transdermal, rectal, nasal, topical (including buccal and sublingual), vaginal, intraperitoneal, intrapulmonary, and intranasal. For local immunosuppressive treatment, the compounds can be administered to the lesion, including perfusion or other contact of the graft with an inhibitor prior to transplantation. It should be understood that the preferred route may vary, for example, depending on the condition of the recipient. When the compound is administered orally, it can be formulated as a pill, capsule, tablet, and the like. with a pharmaceutically acceptable carrier or excipient. When the compound is administered parenterally, it can be formulated with a pharmaceutically acceptable parenteral carrier and in a unit dosage injectable form, as detailed below.

A dose for treating humans can range from about 10 mg to about 1000 mg of a compound of Formula I. A typical dose can range from about 100 mg to about 300 mg of a compound. The dose can be administered once a day (quater in die, QID), twice a day (bis in die, BID), or more often, depending on the pharmacokinetic and pharmacodynamic properties, including absorption, distribution, metabolism, and excretion of the particular compound. In addition, toxicity factors can influence the dosage and administration schedule. For oral administration, pills, capsules, or tablets can be taken daily or less frequently over a period of time. The scheme can be repeated over a number of treatment cycles.

Methods of Treating Compounds of Formula I

The compounds of formula I according to this invention can be used to treat a human or animal suffering from a disease or disorder resulting from abnormal growth, function or behavior of cells associated with USP7, such as immune disorder, cardiovascular disease, viral infection, inflammation, metabolic / endocrine disorder or neurological disorder that can be treated according to a method comprising administering a compound of this invention as defined above. A human or animal suffering from cancer can also be treated in accordance with a method comprising administering a compound of this invention as defined above. The patient's condition can thereby be improved or alleviated.

The methods of this invention also include the treatment of a cancer selected from breast cancer, ovarian cancer, cervical cancer, prostate cancer, testis cancer, urinary tract cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, stomach cancer, skin cancer, keratoacanthoma , lung cancer, squamous cell carcinoma, large cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, adenocarcinoma of the lung, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, carcinoma of the thyroid gland, follicular follicularis, carcinoma nondimensional , seminoma, melanoma, sarcoma, bladder carcinoma, liver and bile duct carcinoma, renal carcinoma, pancreatic cancer, myeloid disorders, lymphoma, hairy cell leukemia, oral cancer, nasopharyngeal cancer, pharyngeal cancer, lip cancer, tongue cancer, thin cancer colon, colon and rectal cancer, colon cancer, ra rectal cancer, brain and central nervous system cancer, Hodgkin's disease, leukemia, bronchial cancer, thyroid cancer, liver and intrahepatic bile duct cancer, hepatocellular cancer, stomach cancer, glioma / glioblastoma, endometrial cancer, melanoma, kidney and renal cancer pelvis, bladder cancer, uterine body cancer, cervical cancer, multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, chronic lymphoid leukemia (CLL), myeloid leukemia, oral and pharyngeal cancer, non-Hodgkin's lymphoma, melanoma the villi of the colon.

Pharmaceutical formulations

In order to use a compound of this invention for the therapeutic treatment of mammals, including humans, it is usually formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition. In accordance with this aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of this invention in combination with a pharmaceutically acceptable diluent or carrier.

A typical formulation is prepared by mixing a compound of this invention and a carrier, diluent or excipient. Suitable carriers, diluents and excipients are well known to those skilled in the art and include materials such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like. The particular carrier, diluent or excipient used will depend on the means and the purpose for which the compound of this invention is used in a given instance. Solvents are generally selected based on solvents recognized by those skilled in the art.

- 86 038835 areas as safe (GRAS) for administration to a mammal. Typically, safe solvents are non-toxic aqueous solvents such as water and other non-toxic solvents that are soluble or miscible with water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycols (e.g., PEG 400, PEG 300), etc. and mixtures thereof. The formulations can also include one or more of buffers, stabilizing agents, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, process modifiers, colors, sweeteners, fragrances, flavors and other known additives to provide an elegant presentation of a drug (i.e., a compound of this invention or a pharmaceutical composition thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a drug).

Formulations can be prepared using standard dissolution and mixing procedures. For example, a drug substance (i.e., a compound of this invention or a stabilized form of a compound (e.g., a complex with a cyclodextrin derivative or other known complexing agent)) is dissolved in a suitable solvent in the presence of one or more of the excipients described above. The compound of this invention is typically formulated into pharmaceutical dosage forms to obtain an easily controlled dosage of the drug and to allow the patient to comply with the prescribed regimen.

The pharmaceutical composition (or formulation) for use can be packaged in various forms depending on the method used to administer the drug. Typically, the product for distribution includes a container containing a pharmaceutical formulation stored therein in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-resistant block to prevent inadvertent access to the contents of the package. In addition, the container is labeled with a label describing the contents of the container. The label may also include appropriate cautions.

Pharmaceutical formulations of the compounds of this invention can be prepared for various routes and types of administration. For example, a compound of Formula I, having the required purity, can optionally be mixed with pharmaceutically acceptable diluents, carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences (1980) 16th edition, Osol, A. Ed.) In the form of a lyophilized formulation, milled powder or an aqueous solution. The composition can be prepared by mixing at room temperature and at an appropriate pH and in the desired degree of purity with physiologically acceptable carriers, i. E. carriers that are non-toxic to recipients at the doses and concentrations used. The pH of the formulation depends primarily on the particular application and the concentration of the compound, but can range from about 3 to about 8. The formulation in an acetate buffer at pH 5 is a suitable embodiment.

The compound can usually be stored as a solid composition, lyophilized composition, or as an aqueous solution.

The pharmaceutical compositions of the invention will be formulated, dosed and administered in such a manner, i. E. in quantity, concentration, queue, course, vehicle, and in a manner consistent with good medical practice. Factors to be considered in this context include the specific disorder to be treated, the specific mammal to be treated, the clinical condition of the individual patient, the cause of the disorder, the site of agent delivery, route of administration, schedule of administration, and other factors known to practitioners. The therapeutically effective amount of a compound to be administered will be determined by such considerations and is the minimum amount needed to ameliorate or treat a hyperproliferative disorder.

As a general proposal, the starting pharmaceutically effective amount of the inhibitor administered parenterally in a single dose will be in the range of about 0.01-100 mg / kg, namely about 0.1-20 mg / kg body weight per day, with a typical starting range the compound used is from 0.3 to 15 mg / kg / day.

Acceptable diluents, carriers, excipients, and stabilizers are non-toxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentresanol; and m-centresanol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as ethylenediaminetetraacetic acid

- 87 038835 (EDTA); sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (for example, Zn-protein complexes); and / or non-ionic surfactants such as TWEEN ™, PLURONICS ™ or polyethylene glycol (PEG). Active pharmaceutical ingredients can also be enclosed in microcapsules obtained, for example, using coacervation or interfacial polymerization techniques, for example, hydroxymethylcellulose or gelatin microcapsules and polymethyl methacrylate microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microchips and nanocapsules) or in macroemulsions. Such techniques are described in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

Sustained release formulations of the compounds of Formula I can be made. Suitable examples of sustained release formulations include semi-permeable matrices of solid hydrophobic polymers containing a compound of Formula I, wherein the matrices are present as shaped articles, eg films, or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly (2-hydroxyethyl methacrylate) or polyvinyl alcohol), polylactides (US 3773919), copolymers of L-glutamic acid and γ-ethyl-L glutamate, non-degradable ethylene vinyl acetate, degradable acid glycolic acid such as LUPRON DEPOT ™ (injection microspheres consisting of a copolymer of lactic acid and glycolic acid and leuprolide acetate) and poly-O - (-) - 3-hydroxybutyric acid.

These formulations include those formulations that are suitable for the routes of administration described herein. The formulations can conveniently be presented in unit dosage form and can be prepared by any of the methods well known in the pharmaceutical art. Techniques and formulations can generally be found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA, USA). Such methods include the step of associating the active ingredient with the carrier, which consists of one or more accessory ingredients. Typically, the compositions are prepared by uniformly and directly associating the active ingredient with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product.

Formulations of a compound of Formula I suitable for oral administration can be prepared as discrete units such as pills, capsules, cachets or tablets, each containing a predetermined amount of a compound of Formula I. Compressed tablets can be prepared by compressing the active an ingredient in free flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets may optionally be coated or may be divisible and optionally formulated so as to provide slow or controlled release of the active ingredient therefrom. Tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules such as gelatin capsules, syrups or elixirs can be prepared for oral use. Oral compositions of the compounds of Formula I may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more agents, including sweeteners, flavors, colors and preservatives, to to provide the drug with a pleasant taste. Acceptable are tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch, gelatin or gum arabic; and lubricating agents such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or may be coated using known techniques, including microencapsulation, to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained release over a longer period. For example, a retention material such as glyceryl monostearate or glyceryl distearate, alone or with wax, can be used.

For the treatment of the eyes or other external tissues, such as the mouth and skin, the compositions are preferably applied as an ointment or cream containing the active ingredient (s) in an amount of, for example, 0.075 to 20% by weight. When formulated as an ointment, the active ingredients can be used with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients can be formulated as a cream with an oil-in-water cream base. If desired, the aqueous phase of the cream base may include a polyhydric alcohol, i. E. alcohol having two or more two hydroxyl groups such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerin and polyethylene glycol (including PEG 400); and mixtures thereof. Topical formulations if desired

- 88 038835 may include a compound that enhances absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulfoxide and related analogs. The oil phase of the emulsions of this invention can be formulated from known ingredients in a known manner. Although the phase may contain only an emulsifier, it desirably contains a mixture of at least one emulsifier with either fat or oil or both fat and oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both oil and fat. Together, the emulsifier (s), with or without a stabilizer, forms the so-called emulsifying wax, and the wax, together with oil and fat, forms the so-called emulsifying ointment base, which forms the oily dispersed phase of the cream formulations. Emulsifiers and emulsion stabilizers suitable for use in the compositions of the invention include Tween® 60, Span® 80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, and sodium lauryl sulfate.

Aqueous suspensions of compounds of formula I contain the active materials in admixture with excipients suitable for the preparation of aqueous suspensions. Such excipients include a suspending agent such as sodium carboxymethylcellulose, croscarmellose, povidone, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic, and dispersing or wetting agents such as naturally occurring phosphate oxides fatty acid (e.g. polyoxyethylene stearate), condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g. heptadecaethyleneoxycethanol), condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol anhydride (e.g. polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl phydroxybenzoate, one or more colorants, one or more flavoring agents, and one or more sweeteners such as sucrose or saccharin ...

Pharmaceutical compositions of the compounds of Formula I may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. This suspension can be formulated in accordance with the prior art using suitable 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 diluent or solvent acceptable for parenteral administration, such as a solution in 1,3-butanediol, or may be formulated as a lyophilized powder. Acceptable carriers and solvents that can be used include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, non-volatile oils can traditionally be used as a solvent or suspending medium. For this, any soft non-volatile oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can be similarly used in the preparation of injectables.

The amount of active ingredient that can be combined with the carrier material to provide a unit dosage form will vary depending on the patient and the particular mode of administration. For example, a sustained release formulation for oral administration to humans may contain from about 1 to 1000 mg of active material mixed with an appropriate and convenient amount of carrier material, which may range from about 5 to about 95% of the total volume of the compositions (by mass). The pharmaceutical composition can be formulated to provide readily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion may contain from about 3 to 500 μg of active ingredient per ml of solution so that infusion can occur in a suitable volume at a rate of about 30 ml / h.

Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatic agents, and solutes that render the composition isotonic with the blood of the intended recipient; as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents.

Formulations suitable for topical administration to the eye also include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous vehicle for the active ingredient. The active ingredient is preferably present in such compositions at a concentration of from about 0.5 to 20% by weight, for example from about 0.5 to 10% by weight, for example about 1.5% by weight.

Formulations suitable for topical administration to the oral cavity include lozenges containing the active ingredient in a flavored base, usually sucrose and gum arabic or tragacanth; lozenges containing the active ingredient in an inert base such as gelatin and glycerin or sucrose and gum arabic; and mouthwashes containing the active ingredient in a suitable liquid carrier.

Formulations for rectal administration may be presented as suppositories with a suitable base containing, for example, cocoa butter or salicylate.

Formulations suitable for intrapulmonary or nasal administration have a particle size of, for example, in the range of 0.1 to 500 microns (including particle sizes in the range of 0.1 to 500 microns in increments of microns, for example, 0.5, 1, 30 , 35 μm, etc.), which are administered by rapid inhalation through the nasal passage or by inhalation through the mouth so that they reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration can be prepared according to conventional methods and can be delivered with other therapeutic agents, such as compounds currently used in the treatment or prevention of the disorders described below.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing, in addition to the active ingredient, such carriers as will be suitable and known in the art.

The formulations can be packaged in unit dose containers or in multi-dose containers, such as sealed ampoules and vials, and can be stored in a lyophilized (freeze-dried) state requiring only the addition of a sterile liquid carrier, such as water, for injection just prior to use. Solutions and suspensions for injection are prepared immediately prior to administration from sterile powders, granules and tablets of the type previously described. Preferred unit dosage formulations contain a daily dose of the active ingredient, or one daily sub-dose, as described above, or an appropriate portion thereof.

This invention also provides compositions for veterinary use containing at least one active ingredient as described above, together with a veterinary carrier. Veterinary carriers are substances suitable for administration of the composition, and can be solid, liquid or gaseous materials that are inert or acceptable in veterinary medicine and are compatible with the active ingredient. These veterinary compositions can be administered parenterally, orally, or by any other route desired.

Combination therapy

The compounds of Formula I can be used alone or in combination with additional therapeutic agents to treat a disease or disorder described herein, such as inflammation or a hyperproliferative disorder (eg, cancer). In some embodiments, a compound of Formula I is combined in a pharmaceutical combination formulation or dosed in a combination therapy regimen with an additional second therapeutic compound that has anti-inflammatory and anti-hyperproliferative properties, or is used to treat inflammation, immune response disorder, or hyperproliferative disorder (e.g., cancer). The additional therapeutic agent can be a Bcl-2 inhibitor, a JAK inhibitor, a PI3K inhibitor, an mTOR inhibitor, an anti-inflammatory agent, an immunomodulatory agent, a chemotherapeutic agent, an apoptosis enhancer, a neurotropic factor, an agent for treating cardiovascular disease, an agent for treating liver disease, an antiviral an agent, an agent for treating blood diseases, an agent for treating diabetes, and an agent for treating immunodeficiency disorders. The second therapeutic agent may be an anti-inflammatory NSAID (nonsteroidal anti-inflammatory agent). The second therapeutic agent can be a chemotherapeutic agent. The second compound of the pharmaceutical combination formulation or dosage regimen preferably has additional activities to the compound of formula I such that they do not adversely affect each other. Such compounds are preferably present in combination in amounts that are effective for a given purpose. In one embodiment, a composition of this invention comprises a compound of Formula I, or a stereoisomer, tautomer, solvate, metabolite, or pharmaceutically acceptable salt or prodrug thereof, in combination with a therapeutic agent such as an NSAID.

Combination therapy can be carried out in the form of a concurrent or sequential regimen. When administered sequentially, the combination can be administered via two or more administrations. Combined administration includes co-administration using separate formulations or a single pharmaceutical formulation and sequential administration in any order, where preferably there is a period of time when both (or all) active agents simultaneously exert their biological activities.

Suitable dosages for any of the above co-administered agents are those currently used and may be reduced by the combined action (synergy) of the newly identified agent and other therapeutic agents or procedures.

Combination therapy can provide synergy and synergistic effects, i.e. when the effect achieved when the active ingredients are used together is greater than the sum of the effects that are obtained when the compounds are used separately. A synergistic effect can be obtained if the active ingredients (1) are combined in one preparation and are administered or dosed simultaneously in the form of a combined unit dosage form; (2) delivered in turns or in parallel as separate trains; or (3) introduced in some other way. When an alternative therapy is used, a synergistic effect can be achieved if the compounds are administered or delivered sequentially, i. E. as different injections in separate syringes, separate tablets or capsules, or separate infusions. Typically, in alternative therapy, an effective dose of each active ingredient is administered sequentially, i. E. in order, while in combination therapy, effective doses of two or more active ingredients are administered together.

In a specific embodiment of therapy, a compound of Formula I, or a stereoisomer, tautomer, solvate, metabolite, or pharmaceutically acceptable salt or prodrug thereof, can be combined with other therapeutic, hormonal agents or antibodies such as those described herein, as well as with surgical treatment and radiation therapy. Thus, combination therapy in accordance with this invention comprises the administration of at least one compound of Formula I, or a stereoisomer, tautomer, solvate, metabolite, or pharmaceutically acceptable salt or prodrug thereof, as well as the use of at least one other method of treating cancer. The amounts of the compound (s) of Formula I and the other pharmaceutically active therapeutic agent (s) and the relative periods of administration are selected so as to achieve the desired combined therapeutic effect.

In some embodiments, a compound of Formula I, or a pharmaceutically acceptable salt thereof, is used in combination with an aromatase inhibitor, a phosphoinositide 3-kinase (PI3K) / mTOR pathway inhibitor, a CDK 4/6 inhibitor, a HER-2 inhibitor, an EGFR inhibitor, a PD-1 inhibitor, poly-ADP-ribose polymerase (PARP) inhibitor, histone deacetylase (HDAC) inhibitor, HSP90 inhibitor, VEGFR inhibitor, AKT inhibitor, chemotherapy, or any combination thereof.

In some embodiments, a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, is administered in combination with a therapeutic agent selected from paclitaxel, anastrozole, exemestane, cyclophosphamide, epirubicin, fulvestrant, letrozole, gemcitabine, trastuzumab (HERCEPTIN®, Trastuzumab) emtansine (KADCYLA®, Genentech), pegfilgrastim, filgrastim, tamoxifen, docetaxel, toremifene, vinorelbine, capecitabine, and ixabepilone.

In some embodiments, a compound of Formula I, or a pharmaceutically acceptable salt thereof, is used in combination with hormone blocking therapy, chemotherapy, radiation therapy, monoclinal antibodies, or combinations thereof.

Hormone blocking therapy involves the use of agents that block estrogen production or block estrogen receptors. In some embodiments, hormone blocking therapy includes the use of estrogen receptor modulators and aromatase inhibitors. Estrogen receptor modulators include triphenylethylene derivatives (eg, tamoxifen, toremifene, droloxifene, 3-hydroxytamoxifen, idoxifene, TAT-59 (phosphorylated derivative of 4hydroxytamoxifen), and GW5638 (carboxylic acid derivative of tamoxifen)); non-steroidal estrogen receptor modulators (eg, raloxifene, LY353381 (SERM3) and LY357489); steroid estrogen receptor modulators (eg ICI-182,780). Aromatase inhibitors include steroidal aromatase inhibitors and non-steroidal aromatase inhibitors. Steroid aromatase inhibitors include, but are not limited to, exemestane. Non-steroidal aromatase inhibitors include, but are not limited to, anastrozole and letrozole.

In some embodiments, a compound of Formula I, or a pharmaceutically acceptable salt thereof, is administered in combination with a CDK 4/6 inhibitor. In some embodiments, the CDK 4/6 inhibitor is palbociclib (PD-0332991), ribociclib (LEE011), or LY283519. In some embodiments, the CDK4 / 6 inhibitor is LEE011. In some embodiments, ribocyclib (LEE011) is administered at a dose of from about 10 mg per day to about 1000 mg per day. In some embodiments, LEE011 is administered at a dose of about 400 mg per day, about 500 mg per day, or about 600 mg per day. In some embodiments, the daily dose of LEE011 is administered orally. In some embodiments, the daily dose of ribociclib (LEE011) is orally administered once a day for three weeks, followed by a week of medication rest during which no ribociclib (LEE011) is administered.

In some embodiments, a compound of Formula I, or a pharmaceutically acceptable salt thereof, is administered in combination with an inhibitor of the phosphoinositide 3-kinase (PI3K) / mTOR pathway. In some embodiments, the phosphoinositide-3-kinase (PI3K) / mTOR pathway inhibitor is everolimus, temsirolimus, BEZ235 (dactolisib), BYL719 (alpelisib), GDC0032 (tazelisib), BKM120 (buparlisib, GDCDC- BGT226) 0980 (apitolisib), GDC0941 (pictilisib), INK128 (MLN0128), INK1117, OSI-027, CC-223, AZD8055, SAR245408, SAR245409, PF04691502, WYE125132, GSK2126458, GSF86936771 , ZSTK474, Cal101 (idealisib), PWT33597, CU-906, AZD-2014 or CUDC-907. In some embodiments, the phosphoinositide 3-kinase (PI3K) / mTOR pathway inhibitor is everolimus. In some embodiments, everolimus is administered at a dose of from about 1 mg per day to about 20 mg per day. In some embodiments, everolimus

- 91 038835 is administered at a dose of about 2.5 mg per day, about 5 mg per day, or about 10 mg per day. In some embodiments, the daily dose of everolimus is administered once a day. In some embodiments, the phosphoinositide 3-kinase (PI3K) / mTOR pathway inhibitor is BKM120 (buparlisib). In some embodiments, BKM120 (buparlisib) is administered at a dosage of about 5 mg per day to about 500 mg per day. In some embodiments, BKM120 is administered at a dose of from about 50 mg per day to about 100 mg per day. In some embodiments, BKM120 is administered at a dose of about 100 mg per day. In some embodiments, the daily dose of BKM120 is administered once a day. In some embodiments, the phosphoinositide 3-kinase (PI3K) / mTOR pathway inhibitor is BYL719. In some embodiments, BYL719 is administered at a dose of from about 25 mg per day to about 1000 mg per day. In some embodiments, BYL719 is administered at a dose of about 250 mg per day, or about 350 mg per day. In some embodiments, the daily dose of BYL719 is administered once a day.

Metabolites of Compounds of Formula I

Also within the scope of this invention are the in vivo metabolic products of Formula I as described herein. Such products can be formed, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic degradation, and the like. of the introduced compound. Accordingly, the invention includes metabolites of compounds of Formula I, including compounds obtained by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

Metabolic products are usually detected by preparing a radiolabeled (e.g. ¹⁴ C or 3H) compound of this invention, parenterally administering it at a detectable dose (e.g., greater than about 0.5 mg / kg) to an animal such as a rat, mouse, marine mumps, monkeys, or humans, leaving enough time for metabolism to occur (usually about 30 s to 30 hours), and excretion of its products from urine, blood or other biological samples. These products are easy to isolate because they are labeled (others are isolated using antibodies capable of binding epitopes retained in the metabolite). Metabolic structures are determined by conventional means, for example by MS, LC / MS or NMR (nuclear magnetic resonance) analyzes. Typically, metabolite analysis is performed in the same manner as conventional drug metabolism studies well known to those of skill in the art. Metabolic products, as long as they are not found in vivo, can be used in diagnostic assays for therapeutic dosing of the compounds of this invention.

Manufactured products

In another embodiment, this invention provides a product of manufacture, or a kit containing materials that can be used to treat the diseases and disorders described above. In one embodiment, the kit contains a container containing a compound of Formula I, or a stereoisomer, tautomer, solvate, metabolite, or pharmaceutically acceptable salt or prodrug thereof. The kit may also contain a label or insert included in the package or connected to the container. The term package insert is used to denote instructions usually included in commercial packages of therapeutic products that contain information on indications, use, dosage, administration, contraindications and / or warnings regarding the use of such therapeutic products. Suitable containers include, for example, bottles, vials, syringes, blister packs, etc. The container can be made of various materials such as glass or plastic. The container may contain a compound of Formula I or a composition thereof that is effective in treating the condition and may have a sterile port (eg, the container may be an intravenous solution bag or a vial having a stopper pierced by a hypodermic needle). At least one active agent in the composition is a compound of Formula I. The label or package insert indicates that the composition is used to treat a selected condition such as cancer. In addition, the label or package insert may indicate that the patient being treated has a disorder such as a hyperproliferative disorder, neurodegeneration, cardiac hypertrophy, pain, migraine, or neurotraumatic disease or event. In one embodiment, the label or insert indicates that a composition comprising a compound of Formula I can be used to treat a disorder resulting from abnormal cell growth. The label or package insert may also indicate that the composition can be used to treat other disorders. Alternatively or additionally, the product of manufacture may also contain a second container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution. It can also include other materials that are commercially desirable and user-friendly, including other buffers, diluents, filters, needles, and syringes.

The kit may also contain directions for administering a compound of Formula I and a second pharmaceutical formulation, if present. For example, if the kit contains a first composition containing a compound of Formula I and a second pharmaceutical composition, the kit may also include instructions for simultaneous, sequential, or separate administration of the first and second pharmaceutical compositions to a patient in need thereof.

In another embodiment, the kits are suitable for delivering solid oral forms of a compound of Formula I, such as tablets or capsules. Such a kit preferably comprises a plurality of unit dosages. Such kits may include a dosage card listed in the order of intended use. An example of such a kit is a blister pack. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms. If desired, a memorization aid may be included, for example in the form of numbers, letters or other markings, or with a calendar insert indicating the days in the treatment regimen on which doses can be administered.

In one embodiment, the kit may include (a) a first container with a compound of Formula I contained therein; and optionally (b) a second container with a second pharmaceutical composition contained therein, wherein the second pharmaceutical composition contains a second compound with antihyperproliferative activity. Alternatively or additionally, the kit may also contain a third container containing a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BWFI), phosphate buffered saline, dextrose solution, and Ringer's solution. It can also include other materials that are commercially desirable and user-friendly, including other buffers, diluents, filters, needles, and syringes.

In some other embodiments, where the kit contains a composition of Formula I and a second therapeutic agent, the kit may contain a container for containing the individual compositions, for example, a split vial or a split foil pouch; however, individual compositions can also be contained in a single undivided container. Typically, the kit contains instructions for the administration of the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (eg, oral and parenteral), at different dosage intervals, or when the titration of the individual components of the combination is desired by the attending physician.

Preparation of compounds of formula I

Compounds of formula I can be synthesized by synthetic routes that include processes analogous to those well known in the chemical arts, especially in light of this disclosure, and those described for other heterocycles in Comprehensive Heterocyclic Chemistry II, Editors Katritzky and Rees. Elsevier, 1997, for example, vol. 3; Liebigs Annalen der Chemie, (9): 1910-16, (1985); Helvetica Chimica Acta 41: 1052-60, (1958); Arzneimittel-Forschung, 40 (12): 1328-31, (1990), each of which is specifically incorporated by reference. Starting materials are generally available from commercial sources such as Aldrich Chemicals (Milwaukee, WI, USA), or can be readily prepared using methods well known to those of skill in the art (e.g., prepared by methods generally described in Louis F Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1-23, Wiley, NY (1967-2006 ed.), Or in the Beilsteins Handbuch der organischen Chemie, 4, Aufl.ed. Springer-Verlag, Berlin, including additions (also available through the Beilstein online database).

Synthetic chemical transformations and protecting group (addition and removal of protecting groups) techniques used in the synthesis of compounds of Formula I, as well as necessary reagents and intermediates, are well known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); TW Greene and PrM Wuts, Protective Groups in Organic Synthesis, 3 ^rd Ed, John Wiley and Sons (1999).; and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and their subsequent editions.

Compounds of formula I can be prepared individually or as compound libraries containing at least 2, for example 5 to 1000 compounds or 10 to 100 compounds. Libraries of compounds of Formula I can be prepared by a combinatorial split and mix approach or by multiple parallel synthesis using solution or solid phase chemical reactions using procedures known to those of skill in the art. Thus, according to another aspect of the present invention, there is provided a compound library comprising at least two compounds or pharmaceutically acceptable salts thereof.

The examples provided herein provide illustrative methods for preparing compounds of Formula I. Those skilled in the art will appreciate that other synthetic routes may be used to synthesize compounds of Formula I. Although specific starting materials and reagents are depicted and discussed in the drawings and examples, they can be easily replaced with other starting materials and reagents to obtain a variety of derivatives and / or reaction conditions. In addition, many of the illustrative compounds obtained by the described methods can be further modified in light of this disclosure using conventional chemical reactions well known to those skilled in the art.

In the preparation of compounds of formulas I, it may be necessary to protect distant functional

- 93 038835 groups (eg primary or secondary amine) intermediates. The need for such protection will vary depending on the nature of the distant functional group and on the conditions of the preparation methods. Suitable groups for protecting amino groups include acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection can be easily determined by a person skilled in the art. For a general description of protecting groups and their uses, see T.W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

In processes for the preparation of compounds of formula I, it may be advantageous to separate the reaction products from each other and / or from the starting materials. Desired products from each step or series of steps are separated and / or purified to the desired degree of homogeneity using techniques known in the art. Typically, such separations include multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can include any number of methods, including, for example, reversed phase and normal phase; exclusive; ion exchange; methods and apparatus for high, medium and low pressure liquid chromatography; small-scale analytical; simulated moving bed (SMB) chromatography and preparative thin layer or thick layer chromatography; and small-scale thin layer and flash chromatography techniques.

Another class of separation methods involves treating a mixture with a reagent selected to bind or otherwise separate the desired product, unreacted starting material, reaction by-product, and the like. Such reagents include adsorbents or absorbents such as activated carbon, molecular sieves, ion exchange media, and the like. Alternatively, the reagents can be acids in the case of a basic material, bases in the case of an acidic material, binding reagents such as antibodies, binding proteins, selective chelating agents such as crown ethers, liquid / liquid ion extraction reagents (LIX), and etc. The choice of appropriate separation methods depends on the nature of the materials used, for example, on the boiling point and molecular weight during distillation and sublimation, on the presence or absence of polar functional groups during chromatography, on the stability of materials in acidic and basic media during multiphase extraction, etc.

Mixtures of diastereomers can be separated into individual diastereomers based on their physicochemical differences using methods well known to those skilled in the art, for example, chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a mixture of diastereomers by reacting with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride), separating the diastereomers, and converting (e.g., by hydrolysis) the individual diastereoisomers to the corresponding pure enantiomers. In addition, some of the compounds of this invention may be atropisomers (eg, substituted diaryls) and are considered part of this invention. Enantiomers can also be separated using a chiral HPLC (high performance liquid chromatography) column.

A single stereoisomer, eg an enantiomer, substantially free of its stereoisomer, can be obtained by resolving a racemic mixture using such a method. as the formation of diastereomers using optically active resolving agents (Eliel, E. and Wilen, S. Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994; Lochmuller, CH, (1975) J. Chromatogr., 113 (3): 283-302). Racemic mixtures of chiral compounds of this invention can be separated and isolated using any suitable method, including: (1) formation of ionic, diastereomeric salts with chiral compounds and separation by fractional crystallization or other means, (2) formation of diastereomeric compounds with chiral derivatization reagents , separating diastereomers and converting to pure stereoisomers, and (3) separating substantially pure or enriched stereoisomers directly under chiral conditions. See Drug Stereochemistry, Analytical Methods and Pharmacology, Irving W. Wainer, Ed., Marcel Dekker, Inc., New York (1993).

In method (1), diastereomeric salts can be formed by reacting enantiomerically pure chiral bases such as brucine, quinine, ephedrine, strychnine, α-methyl-β-phenylethylamine (amphetamine) and the like, with asymmetric compounds bearing an acidic functional a group such as carboxylic acid and sulfonic acid. Diastereomeric salts can be induced to separate by fractional crystallization or ion chromatography. For the separation of optical isomers of amino compounds, the addition of chiral carboxylic or sulfonic acids such as camphorsulfonic acid, tartaric acid, mandelic acid or lactic acid can lead to the formation of diastereomeric salts.

Alternatively, in method (2), the substrate to be separated is reacted with one enantiomer of the chiral compound to form a diastereomeric pair (E. and Wilen, S. Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., 1994, p. 322). Diastereomeric compounds can be formed by the reaction of asymmetric compounds with enantiomerically pure chi

- 94 038835 with other reagents for derivatization, such as menthyl derivatives, followed by separation of diastereomers and hydrolysis to form a pure or enriched enantiomer. The method for determining optical purity involves the preparation of chiral esters such as a menthyl ester, for example (-) menthyl chloroformate, in the presence of a base or a Mosher ester, a-methoxy-a- (trifluoromethyl) phenyl acetate (Jacob III. J. Org. Chem. (1982) 47: 4165), racemic mixture, and analysis of the ¹ H NMR spectrum for the presence of two atropisomeric enantiomers or diastereomers. Stable diastereomers of atropisomeric compounds can be separated and isolated by normal phase and reversed phase chromatography followed by separation methods for atropisomeric naphthyl isoquinolines (WO 96/15111). In method (3), the racemic mixture of the two enantiomers can be separated by chromatography using a chiral stationary phase (Chiral Liquid Chromatography (1989) WJ Lough, Ed., Chapman and Hall, New York; Okamoto, J. Chromatogr., (1990) 513 : 375-378). Enriched or purified enantiomers can be distinguished using methods used to distinguish other chiral molecules with asymmetric carbon atoms, such as optical rotation and circular dichroism.

Compounds of Formula I can be prepared according to the general procedures from Schemes 1-7.

Scheme 1

Scheme 1 shows parahydroxybenzaldehyde intermediate 1 reacting with tert-butyl 3-iodazetidine-1-carboxylate to form an exemplary tert-butyl 3- (4formylphenoxy) azetidine-1-carboxylate intermediate 2. Illustrative intermediate 1 is 2, 6-difluoro-4-hydroxybenzaldehyde. Cyclization of 2 with bicyclic amines 3 gives the tricyclic tetrahydropyrido [3,4-b] indol-1-yl-azetidine intermediate 4. Acid deprotection of 4 and alkylation of 5 gives the tricyclic tetrahydropyrido [3,4-b] indol-1-yl -azetidine intermediate 6.

- 95 038835

Scheme 2

Scheme 2 shows how paraiodobenzaldehyde intermediates 7, such as 2,6difluoro-4-iodobenzaldehyde, cyclize with bicyclic amines 3 to form the tricyclic tetrahydropyrido [3,4-b] indol-1-yl-iodophenyl intermediate 8. Reaction 8 with alcohol 9 gives the tricyclic tetrahydropyrido [3,4-b] indol-1-yl intermediate 10.

Scheme 3

Scheme 3 shows the reaction of amine 11 with an alkylating reagent in which the remaining group can be iodide or bromide or triflate, resulting in intermediate 12. Alternatively, amine 11 can also react with an aldehyde or ketone to form intermediate 12 via a reducing reaction amination. Condensation of intermediate 12 with an aldehyde then gives intermediate 13. The iodide or bromide on the X ¹ group in Cy can then combine with an alcohol or amine or sulfide or olefin via a Pd- or Cu-catalyzed Ullmann or Buchwald or Heck reaction to provide the target compound 14. Alternatively, the protected phenol (OP) on the Cy group can be deprotected and the resulting phenol can then be combined with the alcohol via the Mitsunobu reaction. Alternatively, phenol can also be alkylated, can be reacted with iodide, or bromide, or chloride, or triflate, or mesylate to provide the tricyclic tetrahydropyrido [3,4-b] indol-1-yl intermediate 14.

- 96 038835

Scheme 4

Scheme 4 shows that the Pictet-Spengler cyclization of amine 11 with an aldehyde leads to the formation of intermediate 15, in which X ¹ is iodide or bromide. The reaction of amine 15 with acid chloride gives amide 16. The iodide or bromide group X ¹ on Cy can then combine with an alcohol, or amine, or sulfide, or olefin via a Pd- or Cu-catalyzed Ullmann, or Buchwald, or Heck reaction to form an intermediate product 17. Alternatively, the protected phenol (OP) on the Cy group at 16 can be deprotected and the resulting phenol can then be combined with the alcohol via the Mitsunobu reaction to form 17. Alternatively, the phenol (OH) can be alkylated, can be reacted with iodide , or bromide, or chloride, or triflate, or mesylate, to form the tricyclic tetrahydropyrido [3,4-b] indol-1-yl intermediate 17.

Scheme 5

Scheme 5 shows that amine 15 can react with sulfonyl chloride to form sulfonamide 18, which can be converted via the Pd- or Cu-catalyzed Ullmann, or Buchwald, or Heck reaction, or via Mitsunobu or alkylation reactions to the tricyclic tetrahydropyrido [3,4-b ] indol-1-yl-sulfonamide intermediate 19.

Scheme 6 □ 3 shield

Hr ⁴ xr ⁵ \ z ^{r (R6) m} \ ^ / νη x = i, OTf, Br ^{<R6) m} vr 1nu

L H / \ or reductive kJAA 71

Y amination with aldehyde _R θΥ

X ¹ or ketone \ ι

Scheme 6 shows that amine 15 can be reacted with an alkylating agent (R5-X) to form intermediate 13. Alternatively, amine 15 can be reacted with an aldehyde or ketone and a reducing agent such as sodium cyanoborohydride to provide intermediate 13.

- 97 038835

Scheme 7

Scheme 7 shows a general route for the synthesis of tryptamine 23. Substituted indole 20 is converted to aldehyde 21 under the conditions of the Vilsmeier reaction. The aldol reaction of aldehyde 21 with nitroethane affords compound 22. Reduction of 22 with lithium aluminum hydride then affords tryptamine 23.

Examples of

Example 101 (1 R, 3R) -1 - (2,6-difluoro-4 - ((1 - (3-fluoropropyl) azetidin-3-yl) oxy) phenyl) -2- (2-fluoro-2methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 101.

Step 1: 3- (3,5-difluoro-4-formylphenoxy) azetidine-1-carboxylic acid tert-butyl ester 101c

Cesium carbonate (3.09 g , 9.48 mmol) and 1Boc-3-iodazetidine 101b (CAS No. 254454-54-1, 2.68 g, 9.48 mmol). The resulting mixture was heated at 150 ° C in the microwave for 1 h. The reaction mixture was allowed to cool to ambient temperature, the solid was removed by filtration, the filter cake was washed with toluene and the filtrate was concentrated in vacuo. The precipitate was partitioned between EtOAc and water, the organic phase was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was adsorbed onto HMN diatomaceous earth (Isolute®, Biotage) and purified by silica gel chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 30%) to give 101c as a yellow oil (1.10 g, 93%) ...

¹ H NMR (300 MHz, CDCb): δ 10.20 (s, 1H), 6.35 (m, 2H), 4.94-4.86 (m, 1H), 4.34 (ddd, J = 1.1, 6.4, 9.8 Hz, 2H), 4.05-3.98 (m, 2H), 1.45 (s, 9H).

Step 2: (2-fluoro-2-methylnropyl) - [(R) -2- (1H-indol-3-yl) -1-methylethyl] amine 101d

Compound 101d was prepared according to WO 2014/191726, page 78.

Step 3: 3- {3,5-difluoro-4 - [(1R, 3R) -2- (2-fluoro-2-methylnropyl) -3-methyl-2,3,4,9-tetrahydro-1H-βcarboline -1-yl] phenoxy} azetidine-1-carboxylic acid tert-butyl ester 101e

To a solution of (2-fluoro-2-methylpropyl) - [(R) -2- (1H-indol-3-yl) -1-methylethyl] amine 101d, obtained

- 98 038835 according to WO 2014/191726, p. 78 (540 mg, 2.17 mmol), in toluene (8 ml) under argon atmosphere was added 3 (3,5-difluoro-4-formylphenoxy) azetidine-1-carboxylic acid tert-butyl ester 101c (818 mg, 2.61 mmol) and acetic acid (249 μL, 4.34 mmol). The mixture was heated at 80 ° C in a sealed tube for 4 h, protected from light. The reaction mixture was allowed to cool to room temperature (RT) and concentrated in vacuo. The residue was partitioned between ethyl acetate (EtOAc) and saturated sodium bicarbonate solution. The organic phase was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was adsorbed onto HMN diatomaceous earth and purified by silica gel chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 20%) to give 101e as a white with a yellowish solid (1.10 g, 90%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 7.54 - 7.49 (m, 1H), 7.39 (s, 1H), 7.25 - 7.19 (m, 1H), 7.15 - 7.05 (m, 2H), 6.28-6.21 (m, 2H), 5.20 (s, 1H), 4.84-4.76 (m, 1H), 4.33-4 , 24 (m, 2H), 4.02-3.94 (m, 2H), 3.69-3.61 (m, 1H), 3.12-3.02 (m, 1H), 2.84 (dd, J = 15.1, 20.0 Hz, 1H), 2.65 - 2.56 (m, 1H), 2.38 (dd, J = 14.9, 24.7 Hz, 1H), 1.45 (s, 9H), 1.28-1.08 (m, 9H); LCMS: 544.5 [M + H] +.

Step 4: (1R, 3R) -1- [4- (azetidin-3-yloxy) -2,6-difluorophenyl] -2- (2-fluoro-2-methylpropyl) -3-methyl 2,3,4,9 -tetrahydro-1 H-in-carboline 101f

To a mixture of 3- {3,5-difluoro-4 - [(1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1H-βcarboline- 1-yl] phenoxy} azetidine-1-carboxylic acid tert-butyl ester 101e (840 mg, 1.54 mmol) in dichloromethane (10 ml) under argon, TFA (1.75 ml, 23.1 mmol) was added dropwise and the mixture was shaken, protected from light, at RT for 3 h. The reaction mixture was concentrated in vacuo and purified using an SCX-2 cartridge (mobile phase: dichloromethane / methanol 1: 1, then 2N ammonium in methanol). The appropriate fractions were combined and evaporated to give 101f as a white with a yellowish solid (54 mg, 8%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 7.54-7.49 (m, 1H), 7.41 (s, 1H), 7.25-7.20 (m, 1H), 7.13 - 7.07 (m, 2H), 6.30-6.22 (m, 2H), 5.19 (s, 1H), 4.96-4.90 (m, 1H), 3.97-3 , 91 (m, 2H), 3.83-3.78 (m, 2H), 3.71-3.60 (m, 1H), 3.12-3.03 (m, 1H), 2.85 (dd, J = 15.1, 19.6 Hz, 1H), 2.64 - 2.55 (m, 1H), 2.38 (dd, J = 15.1, 25.2 Hz, 1H), 1.82 (br. S, 1H), 1.27-1.07 (m, 9H); LCMS: 442.5 [MH] ^- .

Step 5: to a mixture of (1R, 3R) -1- [4- (azetidin-3-yloxy) -2,6-difluorophenyl] -2- (2-fluoro-2-methylnropyl) -3methyl-2,3,4 , 9-tetrahydro-1H-β-carboline 101f (54 mg, 0.12 mmol) in N, N-dimethylformamide (2 ml) under argon, 1-bromo-3-fluoropropane (16 μl, 0.16 mmol; CAS No. 352-91-0) and ethyldiisopropylamine (12 μl, 0.24 mmol). The reaction mixture was shaken at RT for 48 h, protected from light. The reaction mixture was poured into a mixture of ethyl acetate and water. The organic layer was separated, washed with water and brine, dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by chromatography on silica gel (mobile phase: dichloromethane / methanol, gradient 0 to 5%) and then using a C18 cartridge (acetonitrile, water, formic acid). The appropriate fractions were combined and evaporated to give 101 as a yellow solid (27 mg, 8%).

1H NMR (400 MHz, CDCl3): δ 11.12 (brs., 1H), 8.27 (s, 1.3H, formic acid), 7.53-7.47 (m, 2H), 7.24 - 7.20 (m, 1H), 7.13 -7.08 (m, 2H), 6.31-6.25 (m, 2H), 5.20 (s, 1H), 4.96-4 , 89 (m, 1H), 4.56 (dd, J = 5.6, 5.6 Hz, 1H), 4.44 (dd, J = 5.6, 5.6 Hz, 1H), 4, 33 - 4.24 (m, 2H), 3.64 (dd, J = 4.8, 11.1 Hz, 1H), 3.49 - 3.47 (m, 1H), 3.07 - 2, 97 (m, 3H), 2.84 (dd, J = 15.0, 20.3 Hz, 1H), 2.64-2.58 (m, 1H), 2.38 (dd, J = 15, 0.24.5Hz, 1H), 1.99 1.83 (m, 2H), 1.27-1.08 (m, 9H); LCMS: 504.3 [M + H] ⁺ .

Example 102 (1 R, 3R) -1 - (2,6-difluoro-4- (2- (3- (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -2- (2-fluoro-2methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 102.

Step 1: (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro1H-β -carboline 102b

- 99 038835

To a solution of (2-fluoro-2-methylpropyl) - [(R) -2- (1H-indol-3-yl) -1-methylethyl] amine 101d, prepared according to WO 2014/191726, page 78, (50 mg , 0.20 mmol), 2,6-difluoro-4-iodobenzaldehyde 102a (CAS No. 1160573-10-3, 65 mg, 0.24 mmol) was added in toluene (170 μl) under argon, followed by acetic acid ( 23 μL, 0.40 mmol). The resulting mixture was shaken at 80 ° C in a sealed tube for 5 h, then left to cool to RT. The mixture was purified on an SCX-2 cartridge (mobile phase: dichloromethane / methanol 9: 1, then 2N ammonium in methanol). The appropriate fractions were combined, evaporated and the crude product was purified by silica gel chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 30%) to give 102b as a yellow solid (89 mg, 89%).

1H NMR (400 MHz, CDCl ₃ ): δ 7.54-7.50 (m, 1H), 7.39 (s, 1H), 7.25-7.21 (m, 3H), 7.16 - 7.08 (m, 2H), 5.26 (s, 1H), 3.67-3.60 (m, 1H), 3.06 (ddd, J = 1.5, 4.9, 15.2 Hz, 1H), 2.86 (dd, J = 15.2, 21.5 Hz, 1H), 2.61 (ddd, J = 1.5, 4.4, 15.2 Hz, 1H), 2 , 39 (dd, J = 15.2, 24.0 Hz, 1H), 1.29 - 1.15 (m, 6H), 1.10 (d, J = 6.4 Hz, 3H); LCMS: 497.0 [MH] ^- .

Stage 2: a mixture of (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9tetrahydro-1H- β-carboline 102b (82 mg, 0.16 mmol), 2- (3-fluoromethylazetidin-1-yl) ethanol 102c prepared according to WO 2013/090836, p. 124 (44 mg, 0.33 mmol; CAS No. 1443984 -69-7, WO 2013/090836), copper iodide (6.2 mg, 0.03 mmol) and potassium carbonate (68 mg, 0.49 mmol) in butyronitrile (600 μl) were degassed in three vacuum / argon cycles. The reaction mixture was heated at 135 ° C for 24 h, allowed to cool to room temperature and diluted with ethyl acetate. The solid was removed from the reaction mixture by filtration through celite, and the solid was washed with ethyl acetate. The combined filtrate was washed with water (three times) and brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel (mobile phase: 0-7% methanol / dichloromethane). The appropriate fractions were collected and evaporated to give 102 as a yellow solid (17.2 mg, 21%).

1H NMR (400 MHz, DMSO-dg): δ 10.51 (s, 1H), 7.39 (d, J = 7.3 Hz, 1H), 7.18 (d, J = 7.8 Hz, 1H), 7.01 - 6.91 (m, 2H), 6.64 (d, J = 11.2 Hz, 2H), 5.11 (s, 1H), 4.56 (d, J = 5 , 9 Hz, 1H), 4.44 (d, J = 5.4 Hz, 1H), 3.92 (s, 2H), 3.543.47 (m, 2H), 3.06 - 2.66 (m , 6H), 2.59 - 2.53 (m, 2H, partially under DMSO-dg), 2.40 - 2.27 (m, 2H), 1.25-1.09 (m, 6H), 1 04 (d, J = 6.4Hz, 3H); LCMS: 502.3 [MH] ^- .

Example 103 1 - ((1R, 3R) -1- (2,6-difluoro-4- (2- (3- (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -3-methyl3,4-dihydro -1 H-pyrido [3, 4-b] indol-2 (9H) -yl) -2-methylpropan-1 -one 103.

Step 1: (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-2,3,4,9-tetrahydro-1H-nyrido [3,4-b] indole 103b

(2R) -1- (1H-indol-3-yl) npropan-2-amine 103 a (710 mg, 3.67 mmol), then 2,6-difluoro-4-iodobenzaldehyde (1 , 1 g, 4.03 mmol) and acetonitrile (2.6 ml). The reaction was placed under nitrogen and TFA (0.5 ml, 7.0 mmol) was added. The reaction mixture was then heated to 130 ° C in a microwave for 1 hour and then quenched with saturated aqueous NaHCO ₃ . The mixture was extracted with DCM (dichloromethane) (3x100 ml), dried with MgSO ₄ , filtered and concentrated. The crude product was purified by flash chromatography through silica gel (0-100% EtOAc / hexanes) to give 103b (450 mg, 29%).

¹ H NMR (400 MHz, deuterochloroform-d): δ 7.60-7.48 (m, 2H), 7.27 (d, J = 7.3 Hz, 2H), 7.17-7.08 ( m, 2H), 5.63 (s, 1H), 3.45 (dq, J = 12.7, 6.2 Hz, 1H), 2.99 (ddd, J = 15.5, 4.6, 1.3 Hz, 1H), 2.52 (ddd, J = 15.5, 7.3, 1.8 Hz, 1H), 1.29 (d, J = 6.5 Hz, 3H).

Stage 2: 1 - ((1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-3,4-dihydro-1H-pyrido [3,4-b] indole-2 ( 9H) yl) -2-methylpropan-1-one 103 s

To a round bottom vessel (RBF) was added (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-2,3,4,9tetrahydro-1H-pyrido [3,4-b] indole 103b (50 mg, 0.12 mmol) followed by sodium bicarbonate (50 mg, 0.59 mmol) and chloroform (0.8 ml). Added 2-methylpropanoyl chloride (31 mg, 0.2947 mmol) and the reaction mixture was heated to 45 ° C for 1 h.Diisopropylethylamine (Hunig's base, 0.1 ml, 0.59 mmol) was added and the reaction was shaken until LCMS did not show that starting materials were depleted of 100 038835 pans. Was added an aqueous saturated solution (10 ml) of sodium bicarbonate. The reaction mixture was then extracted with DCM (3x50 ml), dried over MgSO4, filtered and concentrated. The crude product was purified on a silica gel flash chromatography column (0-100%

EtOAc / hexanes) to give 103c (51 mg, 88%).

1H NMR (400 MHz, DMSO-d ₆ ): δ 10.74 (s, 1H), 7.46 (d, J = 7.8 Hz, 1H), 7.39 (d, J = 9.2 Hz , 2H), 7.24 (dt, J = 8.0, 1.0 Hz, 1H), 7.01 (dddd, J = 26.4, 8.0, 7.0, 1.2 Hz, 2H ), 6.10 (s, 1H), 4.88-4.71 (m, 1H), 3.17 (dd, J = 14.9, 5.6 Hz, 1H), 3.03 (p, J = 6.6 Hz, 1H), 2.84 (d, J = 15.2 Hz, 1H), 1.12 (d, J = 6.5 Hz, 2H), 1.06 - 0.92 ( m, 6H).

Stage 3: 1 - [(1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-1,3,4,9-tetrahydropyrido [3 , 4-b] indol-2-yl] -2-methylpropan-1-one 103c (51 mg, 0.10 mmol), and then 2- [3- (fluoromethyl) azetidin-1-yl] ethanol 102c, obtained according to WO 2013/090836, page 124 (27 mg, 0.21 mmol), copper iodide (8 mg, 0.04 mmol), potassium carbonate (43 mg, 0.31 mmol). The vial was sealed and butyronitrile (0.7 ml) was added. The reaction mixture was then heated to 135 ° C overnight and then cooled to room temperature. The reaction mixture was then filtered through celite, eluting with EtOAc. The combined filtrate was then concentrated and purified by reverse phase HPLC to give 103 (16 mg, 31%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.48 (s, 1H), 7.51-7.39 (m, 1H), 7.32-7.22 (m, 1H), 7 , 09-6.90 (m, 2H), 6.51 (d, J = 11.0 Hz, 2H), 6.11 (s, 1H), 4.89-4.71 (m, 1H), 4.55 (d, J = 6.1 Hz, 1H), 4.43 (d, J = 6.0 Hz, 1H), 3.94 (q, J = 5.4 Hz, 2H), 3, 59 - 3.38 (m, 2H), 3.24 - 3.18 (m, 2H), 3.04 - 2.97 (m, 2H), 2.87 - 2.74 (m, 4H), 1.12 (d, J = 6.4 Hz, 3H), 0.98 (dd, J = 10.3, 6.7 Hz, 6H); LCMS: 500.3 [M + H] +.

Example 104.1 - ((1R, 3R) -1 - (2,6-difluoro-4- (2- (3 - (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -3 -methyl3,4-dihydro -1 H-pyrido [3, 4-b] indol-2 (9H) -yl) -2-fluoro-2-methylpropan-1 -one 104.

Stage 1: 1 - ((1R, 3R) -1 - (2,6-difluoro-4-iodophenyl) -3-methyl-3,4-dihydro-1 H-pyrido [3, 4-b] indole-2 (9Н) yl) -2-fluoro-2-methylpropan-1-one 104a

F

I

(1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-2,3,4,9-tetrahydro1H-pyrido [3,4-b] indole 103b (100 mg, 0.24 mmol), followed by 2-fluoro-2-methylpropanoyl _{chloride (0.59 ml of a 1M solution in CHCl 3} , obtained from the reaction of the corresponding acid with oxalyl chloride), sodium bicarbonate (99 mg, 1.2 mmol) and chloroform (1.6 ml). The reaction was then heated to 45 ° C for 1 h and then Hunig's base (0.2 ml, 1.2 mmol) was added. The reaction was shaken until LC-MS indicated that starting materials were depleted. The reaction was quenched with saturated aqueous sodium bicarbonate solution. The mixture was then extracted with DCM (3x50 ml), dried with MgSO ₄ , filtered and concentrated. The resulting crude product was purified by silica gel column chromatography (0-100% EtOAC / hexanes) to give 104a (95 mg, 79%).

1H NMR (400 MHz, DMSO / D: δ 7.54-7.31 (m, 3H), 7.28-7.21 (m, 1H), 7.04 (ddd, J = 8.1, 7 , 1, 1.3 Hz, 1H), 6.98 (td, J = 7.5, 7.0, 1.1 Hz, 1H), 6.08 (s, 1H), 5.14 (s, 1H), 3.14 (dd, J = 15.4, 4.6 Hz, 1H), 2.81 (d, J = 15.2 Hz, 1H), 1.51 (dd, J = 35.4 , 21.8 Hz, 6H), 1.17 (dt, J = 3.1 Hz, 3H) LCMS: 513.0 [M + H] ⁺ .

Stage 2: 1 - [(1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-1,3,4,9-tetrahydropyrido [3 , 4-b] indol-2-yl] -2-fluoro-2-methylpropan-1-one 104a (29 mg, 0.056 mmol), followed by 2- [3- (fluoromethyl) azetidin-1-yl] ethanol ( 15 mg, 0.11 mmol), copper iodide (4 mg, 0.023 mmol), potassium carbonate (24 mg, 0.17 mmol) and butyronitrile (0.37 ml). The solution was degassed for 5 min and then heated to 135 ° C overnight. When LCMS showed that starting materials were depleted, the crude mixture was cooled to room temperature, filtered through Celite®. The cellite pad was then washed with EtOAc and the combined filtrate was concentrated and purified by reverse phase HPLC to give 104 (9 mg, 31%).

1H NMR (400 MHz, DMSO-d6, 350K): δ 10.69 (s, 1H), 7.54-7.38 (m, 1H), 7.31-7.17 (m, 1H), 7 , 00 (dtd, J = 24.8, 7.1, 1.2 Hz, 2H), 6.55 (d, J = 12.0 Hz, 1H), 6.03 (s, 1H), 5, 21 - 5.05 (m, 1H), 4.54 (d, J = 6.2 Hz, 1H), 4.42 (d, J = 6.2 Hz, 1H), 3.87 (t, J = 5.4 Hz, 2H), 3.30 - 3.25 (m, 2H), 3.15 (dd, J = 15.3, 4.7 Hz, 1H), 2.96 (t, J = 6.5 Hz, 2H), 2.79 (d, J = 15.1 Hz, 1H), 2.75-2.62 (m, 3H), 1.55 (d, J = 21.8 Hz, 2H), 1.45 (d, J = 21.8 Hz, 2H), 1.15 (d, J = 6.4 Hz, 2H); LCMS: 518.2 [M + H] ⁺ .

Example 105. (1 R, 3R) -1 - (4- (2- (3 - (difluoromethyl) azetidin-1-yl) ethoxy) -2,6-difluorophenyl) -2- (2-fluoro-2methylpropyl) - 3-methyl-2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 105.

Step 1: 2- (3,5-difluoro-4-formylphenoxy) ethyl acetate 105a

- 101 038835

A solution of 2,6-difluoro-4-hydroxybenzaldehyde (CAS No. 532967-21-8, 300 mg, 1.89 mmol) and 2-bromoethyl acetate (CAS No. 927-68-4, 0.22 ml, 2 mmol) in acetonitrile (5 ml) and Ν, Ν-dimethylformamide (1 ml) were heated at 80 ° C for 24 h. Another portion of 2-bromoethyl acetate (0.11 ml, 1 mmol) was added and heating was continued at 80 ° C for another 30 am The reaction mixture was left to cool to ambient temperature. The precipitate was partitioned between EtOAc and saturated sodium bicarbonate solution. The aqueous layer was extracted with additional portions of EtOAc. The combined organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 33%) to give 105a as a white powder (213 mg, 45%).

1H NMR (300 MHz, CDCl ₃ ): δ 10.20 (s, 1H), 6.51 (d, J = 10.4 Hz, 2H), 4.44 (t, J = 4.7 Hz, 2H ), 4.22 (t, J = 4.7 Hz, 2H), 2.11 (s, 3H).

Stage 2: 2- (3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [3 , 4-b] indol-1-yl) phenoxy) ethyl acetate 105 b

To a solution of (2-fluoro-2-methylpropyl) - [(R) -2- (1H-indol-3-yl) -1-methylethyl] amine 101d (213 mg, 0.86 mmol) and 2- (3, 5-difluoro-4-formylphenoxy) ethyl acetate 105a (210 mg, 0.86 mmol) in toluene (1 ml) under argon was added glacial acetic acid (0.1 ml, 1.72 mmol). The vial was sealed and the reaction mixture was heated at 80 ° C for 16 hours. The reaction mixture was allowed to cool to ambient temperature. The residue was partitioned between dichloromethane and saturated sodium bicarbonate solution. The aqueous layer was extracted with additional portions of dichloromethane. The combined organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 20%) to give 105b as a white foam (323 mg, 80%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 7.54-7.49 (m, 1H), 7.38 (s, 1H), 7.24-7.19 (m, 1H), 7.14 - 7.07 (m, 2H), 6.42 (dd, J = 13, 3 Hz, 2H), 5.19 (s, 1H), 4.40 (t, J = 4.7 Hz, 2H) , 4.12 (t, J = 4.7 Hz, 2H), 3.70-3.62 (m, 1H), 3.133.04 (m, 1H), 2.92-279 (dd, J = 19, 15 Hz, 1H), 2.65 - 2.55 (m, 1H), 2.46 - 2.31 (dd, J = 25.0, 15.0 Hz, 1H), 2.10 ( s, 3H), 1.24 (d, J = 11.0 Hz, 3H), 1.17 (d, J = 11.3 Hz, 3H), 1.1 (d, J = 6.5 Hz, 3H).

Stage 3: 2- (3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [3 , 4-b] indol-1-yl) phenoxy) ethanol 105 s

To a solution of 2- (3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [3, 4-b] indol-1-yl) phenoxy) ethyl acetate 105b (320 mg, 0.675 mmol) in THF (tetrahydrofuran) / MeOH (2/1, 6 ml) sodium hydroxide (1N, 4 ml) was added. The reaction mixture was heated at 70 ° C for 45 min. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The precipitate was partitioned between dichloromethane and water. The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo to give 105c as a white foam (264 mg, 91%). LCMS: 431.2 [MH] '.

Stage 4: (1K, 3K) -1- (4- (2-bromoethoxy) -2,6-difluorophenyl) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9tetrahydro -1 H-pyrido [3, 4-b] indole 105d

- 102 038835

Ά-wg

To a solution of 2- (3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [3, 4-b] indol-1-yl) phenoxy) ethanol 105c (130 mg, 0.3 mmol) in DCM (2.5 ml), triphenylphosphine (94 mg, 0.36 mmol) and carbon tetrabromide (120 mg, 0 , 36 mmol). The reaction mixture was shaken at room temperature for 1 hour, then the solvent was removed in vacuo. The crude product was purified by silica gel column chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 20%) to give 105d as a white foam (142 mg, 95%).

1H NMR (300 MHz, CDCl ₃ ): δ 7.54-7.49 (m, 1H), 7.38 (s, 1H), 7.25-7.19 (m, 1H), 7.15- 7.07 (m, 2H), 6.42 (dd, J = 13.0, 3.0 Hz, 2H), 5.20 (s, 1H), 4.24 (t, J = 4.7 Hz , 2H), 3.72-3.59 (m, 3H), 3.12-3.03 (m, 1H), 2.922.79 (dd, J = 19.4, 15.0 Hz, 1H), 2.64 - 2.56 (m, 1H), 2.46 - 2.31 (dd, J = 25.0, 15.0 Hz, 1H), 1.24 (d, J = 12.1 Hz, 3H), 1.17 (d, J = 12 Hz, 3H), 1.10 (d, J = 6.5 Hz, 3H).

Step 5: to a solution of (1R, 3R) -1- (4- (2-bromoethoxy) -2,6-difluorophenyl) -2- (2-fluoro-2-methylpropyl) -3methyl-2,3,4,9 -tetrahydro-1H-pyrido [3,4-b] indole 105d (62 mg, 0.125 mmol) in acetonitrile (1 ml) were added N, N-diisopropylethylamine (0.064 ml, 0.375 mmol) and 3- (difluoromethyl) azetidine hydrochloride ( CAS 1354792-76-9, 27 mg, 0.187 mmol). The reaction mixture was shaken at room temperature for 1 hour, then at 45 ° C for 4 hours. The reaction mixture was allowed to cool to ambient temperature. The precipitate was partitioned between EtOAc and water. The aqueous layer was extracted with additional portions of EtOAc. The combined organic layers were separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography (mobile phase: dichloromethane / methanol, gradient 0 to 2.5%) to give 105 as a white with a yellowish solid (40 mg, 62%).

1H NMR (300 MHz, CDCl ₃ ): δ 7.54 - 7.49 (m, 1H), 7.38 (s, 1H), 7.24 - 7.19 (m, 1H), 7.14 - 7.06 (m, 2H), 6.38 (dd, J = 13.3, 3 Hz, 2H), 6.17-5.76 (dt, J = 56.0, 5.1 Hz, 1H) , 5.18 (s, 1H), 3.90 (t, J = 5.3 Hz, 2H), 3.71 - 3.63 (m, 1H), 3.46 (t, J = 7.8 Hz, 2H), 3.27 (t, J = 6.7 Hz, 2H), 3.13 - 3.04 (m, 1H), 2.92 - 2.79 (m, 3H), 2.64 - 2.55 (m, 1H), 2.45 - 2.30 (dd, J = 25.6, 14.9 Hz, 1H), 1.23 (d, J = 10.3 Hz, 3H), 1.16 (d, J = 12 Hz, 3H); 1.09 (d, J = 6.5 Hz, 3H); LCMS: 520.4 [MH] ^- .

Compounds 106-125 were prepared according to the procedures described herein and characterized by LCMS [M + H] +

Example 126. (1 R, 3R) -1 - (2,6-difluoro-4- (2- (3- (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -3-methyl-2 (methylsulfonyl) - 2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 126.

Step 1: (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-2- (methylsulfonyl) -2,3,4,9-tetrahydro-1Hpyrido [3,4-b ] indole

- 103 038835

In a round-bottom vessel with a volume of 50 ml (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl2,3,4,9-tetrahydro-III-pyrido [3,4-L] indole (50 mg, 0.12 mmol) and chloroform (0.15 M, 0.8 ml). Then N, N-diisopropylethylamine (0.06 ml, 0.35 mmol) and methanesulfonyl chloride (0.014 ml, 0.18 mmol) were added sequentially. The reaction was then heated to 45 ° C and monitored until LCMS indicated that starting materials were depleted. The reaction was cooled to room temperature, quenched by addition of saturated aqueous NH4Cl, extracted with DCM (3x50 mL), dried over MgSO _4, filtered and concentrated. The crude product was purified on a silica gel flash chromatography column eluting with 0-50% iPrOAc / heptanes to give the title compound (40 mg, 68% yield).

1H NMR (400 MHz, DMSOD) δ 10.78 (s, 1H), 7.54 (d, J = 7.9 Hz, 2H), 7.44 (d, J = 7.8 Hz, 1H), 7.22 (d, J = 8.1 Hz, 1H), 7.05 (ddd, J = 8.2, 7.1, 1.2 Hz, 1H), 7.02 - 6.95 (m, 1H), 6.18 (s, 1H), 4.43 (q, J = 5.6, 5.0 Hz, 1H), 3.09 - 2.99 (m, 1H), 2.83 (s , 4H), 1.31 (d, J = 6.6 Hz, 3H). LCMS: 503.0 [M + H] +.

Stage 2: (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-2methylsulfonyl-1,3,4,9-tetrahydropyrido [3,4-b ] indole (40 mg, 0.08 mmol), 2- [3- (fluoromethyl) azetidine-1yl] ethanol (21 mg, 0.16 mmol), copper iodide (6 mg, 0.032 mmol), potassium carbonate (33 mg , 0.24 mmol) and butyronitrile (0.5 ml). The solution was degassed for 5 min and then heated to 135 ° C overnight. When LCMS showed that starting materials were depleted, the reaction mixture was filtered through celite, eluting with EtOAc. The filtrate was concentrated and purified by reverse phase HPLC to give 126 (6 mg, 15% yield).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.74 (s, 1H), 7.43 (d, J = 7.7 Hz, 1H), 7.24 -7.20 (m, 1H) , 7.04 (ddd, J = 8.2, 7.0, 1.4 Hz, 1H), 6.97 (td, J = 7.4, 1.1 Hz, 1H), 6.73-6 , 64 (m, 2H), 6.15 (s, 1H), 4.55 (d, J = 6.2 Hz, 1H), 4.45-4.35 (m, 2H), 3.93 ( t, J = 5.4 Hz, 2H), 3.30 - 3.28 (m, 2H), 3.03 - 2.95 (m, 3H), 2.77 (s, 3H), 2.74 2.65 (m, 4H), 1.33 (dd, J = 6.8, 2.1 Hz, 3H). LCMS: 508.2 [M + H] ⁺ .

Example 145. N- (3,5-difluoro-4 - ((1 R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [ 3,4-b] indol-1-yl) phenyl) -1 - (3-fluoropropyl) azetidine-3-amine 145.

Step 1: (1R, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro1 H- pyrido [3, 4-b] indole _ oUd

Bg

To a solution of (R) -N- (1- (1H-indol-3-yl) propan-2-yl) -2-fluoro-2-methylpropan-1-amine (500 mg, 2.01 mmol) in toluene ( 6 ml) was added 4-bromo-2,6-difluorobenzaldehyde (490 mg, 2.21 mmol) and acetic acid (0.58 ml, 10.2 mmol). The reaction mixture was shaken at 80 ° C for 16 h. After cooling to room temperature, the solution was concentrated and the residue was diluted with EtOAc (40 ml), washed with saturated aqueous NaHCO ₃ (10 ml) and water (20 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel chromatography (solvent gradient: 0-6% EtOAc in petroleum ether) to give the title compound (800 mg, 88%) as a light yellow solid.

1H NMR (400 MHz, CDCl ₃ ) δ 7.53 (d, J = 7.2 Hz, 1H), 7.41 (s, 1H), 7.24 (d, J = 7.2 Hz, 1H) , 7.16 - 7.09 (m, 2H), 7.06 (d, J = 8.0 Hz, 2H), 5.27 (s, 1H), 3.73 - 3.54 (m, 1H ), 3.09-3.05 (m, 1H), 2.95-2.76 (m, 1H), 2.64-2.60 (m, 1H), 2.47-2.33 (m , 1H), 1.30-1.17 (m, 6H), 1.11 (d, J = 6.4Hz, 3H).

Stage 2: tert-butyl-3 - ((3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9- tetrahydro1H-pyrido [3,4-b] indol-1-yl) phenyl) amino) azetidine-1-carboxylate

- 104 038835

A mixture of (1R, 3R) -1- (4-bromo-2,6-difluorophenyl) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro1Н-pyrido [3 , 4-b] indole (from stage 1, 800.0 mg, 1.77 mmol), BINAP (110.4 mg, 0.18 mmol), Pd ₂ (dba) ₃ (162.3 mg, 0.18 mmol), tert-BuONa (511.0 mg, 5.32 mmol) and tert-butyl-3-aminoazetidine-1-carboxylate (457.9 mg, 2.66 mmol) in toluene (10 ml) were shaken at 110 ° C under N2 for 16 hours. The reaction mixture was concentrated and purified on a silica gel column (0-5% methanol in DCM) to give the title compound (900 mg, 94%) as a brown solid.

1H NMR (400 MHz, CDCl ₃ ) δ 7.51 (d, J = 6.4 Hz, 1H), 7.43 (s, 1H), 7.22 (d, J = 8.0 Hz, 1H) , 7.13-7.05 (m, 2H), 5.97 (d, J = 11.2 Hz, 2H), 5.14 (s, 1H), 4.37-4.21 (m, 3H ), 4.20-4.01 (m, 1H), 3.78-3.60 (m, 3H), 3.123.07 (m, 1H), 2.96-2.77 (m, 1H), 2.63-2.57 (m, 1H), 2.48-2.33 (m, 1H), 1.45 (s, 9H), 1.25-1.17 (m, 6H), 1, 10 (d, J = 6.0 Hz, 3H).

Step 3: N- (3,5-difluoro-4 - ((1 R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [ 3, 4-b] indol-1-yl) phenyl) azetidine-3-amine

To a mixture of tert-butyl-3 - ((3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9tetrahydro-1H -nirido [3,4-b] indol-1-yl) phenyl) amino) azetidine-1-carboxylate (from step 2, 0.9 g, 1.66 mmol) in DCM (5 ml) was added TFA (1, 8 ml, 24.88 mmol) at -20 ° C. The resulting mixture was shaken at 0 ° C for 16 h. Aqueous NaHCO ₃ solution (80 ml) was slowly added to the reaction mixture and then the reaction mixture was extracted with DCM (100 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give the title compound (700 mg, 95%) as a brown solid. The crude product was used in the next step without further purification.

Stage 4: to a mixture of N- (3,5-difluoro-4 - ((1R, 3R) -2- (2-fluoro-2-methylpropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indol-1-yl) phenyl) αzetidine-3-amine (from step 3, 700.0 mg, 1.58 mmol) and N, N diisopropylethylamine (613.3 mg, 4.75 mmol) in N, N-dimethylformamide (10 ml) was added 1-bromo-3fluoropropane (223.0 mg, 1.58 mmol) and the reaction mixture was shaken at 10 ° C for 16 h. The reaction mixture was purified on a column (0-10% MeOH in DCM) and further purified by reverse phase chromatography (acetonitrile 66-96% / 0.05% NH4OH in water) to give 145 (280 mg, 35%) as a white solid.

1H NMR (400 MHz, CD ₃ OD) δ 7.38 (d, J = 7.6 Hz, 1H), 7.17 (d, J = 7.6 Hz, 1H), 7.03-6.88 (m, 2H), 6.07 (d, J = 11.6 Hz, 2H), 5.10 (s, 1H), 4.54-4.36 (m, 2H), 4.03-4, 01 (m, 1H), 3.79-3.71 (m, 2H), 3.69-3.65 (m, 1H), 3.04-3.00 (m, 1H), 2.97- 2.91 (m, 2H), 2.87-2.85 (m, 1H), 2.62 (t, J = 7.6 Hz, 2H), 2.58-2.55 (m, 1H) , 2.48 - 2.32 (m, 1H), 1.83 - 1.67 (m, 2H), 1.20 - 1.11 (m, 6H), 1.08 (d, J = 6, 8 Hz, 3H).

Example 154 (S) -3 - ((1 R, 3R) -1 - (2,6-difluoro-4- (2- (3 - (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -3 methyl -3,4-dihydro-1H-pyrido [3,4-b] indol-2 (9H) -yl) -2-fluoro-2-methylpropan-1-ol 154.

Stage 1: dimethyl-2-fluoro-2-methylmalonate

Oh oh

Sodium hydride (1.15 equiv., 21 mmol) was added to an oven dried 500 mL round bottom vessel. The reaction mixture was placed under a nitrogen atmosphere and cooled to 0 ° C. Then added THF (63 ml). To this mixture was added dimethyl 2-methylpropanedioate (5.0 g, 34.2 mmol) dropwise and the reaction mixture was shaken for 30 minutes. Then N-fluorobenzenesulfonimide (1.05 eq., 19.2 mmol) was added in one portion. The reaction mixture was allowed to warm to room temperature and the reaction mixture was solidified such that an additional 50 ml of THF was added. After 1.5 h, the reaction was quenched with aqueous 2N. HCl, diluted with EtOAc (500 ml) and washed 3x200 ml 2N. HCl. The organic phase was separated, dried with MgSO ₄ , filtered and concentrated. The crude white solid was then taken up in 200 ml of heptane, sonicated and filtered through celite. The filtered solid was then washed with 3x200 ml heptane. The combined filtrate was then concentrated to give the crude desired product (3 g, 53% yield) as a yellow oil.

1H NMR (400 MHz, DMSO-d ₆ ) δ 3.32 (s, 6H), 1.18 (d, J = 6.3Hz, 3H).

Step 2: 2-fluoro-2-methylpropane-1,3-diol

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Dimethyl 2-fluoro-2methylpropanedioate (3 g, 18.3 mmol) and THF (90 ml) were added to an oven dried 500 ml round bottom vessel. The reaction mixture was placed under N ₂ and then cooled to 0 ° C. Then a solution of lithium aluminum hydride (1M in THF, 2.75 eq., 50.3 mmol) was added dropwise and the reaction was warmed to room temperature for 1 h. Then the reaction was again cooled to 0 ° C and quenched by the addition of water (2 ml ), and then 15% aqueous NaOH solution (2 ml) and water (4 ml) were added. The suspension was shaken for 15 min, filtered and concentrated to give the crude product (1.4 g, 71% yield).

1H NMR (400 MHz, DMSO-d6) δ 4.85 (t, J = 5.9 Hz, 2H), 3.45 (d, J = 5.9 Hz, 2H), 3.41 (d, J = 5.9 Hz, 2H), 1.22-1.15 (d, 3H).

Step 3: 3- (tert-butyldiphenylsilyloxy) -2-fluoro-2-methylpropan-1-ol

OH OTBDPS

Have

Into an oven-dried 500 ml round-bottom vessel was added 2-fluoro-2-methylpropane-1,3diol (1.47 g, 1.25 eq., 13.6 mmol), followed by imidazole (1.11 g, 1, 5 eq., 16.4 mmol), tert-butylchlorodiphenylsilane (3.0 g, 10.9 mmol) and chloroform (136 ml). The reaction was shaken overnight and quenched by the addition of saturated NH ₄ Cl solution (100 ml). The mixture was extracted with DCM (100 ml), dried with MgSO ₄ , filtered and concentrated. The crude mixture was purified by flash silica gel column chromatography (0-100% iPrOAc / heptanes) to afford the desired product (1.26 g, 33% yield).

1H NMR (400 MHz, DMSO-f) δ 7.68-7.60 (m, 4H), 7.51-7.40 (m, 6H), 4.97 (t, J = 5.8 Hz, 1H), 3.70 (dd, J = 19.4, 1.9 Hz, 2H), 3.52 (ddd, J = 18.5, 5.8, 1.8 Hz, 2H), 1.28 (d, J = 21.8 Hz, 3H), 1.01 (s, 9H).

Step 4: 3- (tert-butyldiphenylsilyloxy) -2-fluoro-2-methylpropyltrifluoromethanesulfonate OTf OTBDPS

Into an oven-dried 500 ml round-bottom vessel was added 3- [tert-butyl (diphenyl) silyl] oxy-2-fluoro-2-methylpropan-1-ol (1.3 g, 3.8 mmol), dichloromethane (63 ml) in nitrogen atmosphere. Then the reaction mixture was cooled to 0 ° C and trifluoromethanesulfonic anhydride (1.27 g, 1.2 eq., 4.5 mmol) was added dropwise. Then the reaction mixture was shaken for 2 h and then washed with 2N. HCl and then saturated NaHCO ₃ solution. The organic phase was separated, then dried with MgSO ₄ and filtered through a pad of silica gel, eluting with DCM. The filtrate was then concentrated to dryness to give the crude desired product (1.8 g, 100% yield) and used in the next step without further purification.

1H NMR (400 MHz, DMSO-d6) δ 7.66-7.58 (m, 4H), 7.56-7.41 (m, 6H), 5.07-4.81 (m, 2H), 3.88 -3.68 (m, 2H), 1.40 (d, J = 21.6 Hz, 3H), 1.01 (s, 9H).

Step 5: n - ((R) -1- (1H-indol-3-yl) propan-2-yl) -3- (tert-butyldiphenylsilyloxy) -2-fluoro-2-methylpropan-1-amine

Into an oven dried 250 ml round-bottom vessel was added (2R) -1- (1H-indol-3yl) propan-2-amine (600 mg, 3.1 mmol), N, N-diisopropylethylamine (0.81 ml, 1 , 5 eq., 4.65 mmol) and 1,4-dioxane (6 ml) and the reaction mixture was placed under a nitrogen atmosphere. Then [3- [tert-butyl (diphenyl) silyl] oxy-2-fluoro-2-methylpropyl] trifluoromethanesulfonate (1.95 g, 1.25 eq., 3.9 mmol) was added and the reaction mixture was heated to 90 ° C. When LCMS indicated consumption of starting materials, the reaction mixture was quenched with saturated aqueous NaHCO ₃ and the mixture was extracted with EtOAc (3x200 ml). The combined organic matter was dried with MgSO ₄ , filtered and concentrated. Purification by flash silica gel column chromatography (0-100% EtOAc / hexanes) afforded the title compound (1.2 g, 77% yield). LCMS: 503.3 [M + H] +.

Step 6: 3 - ((R) -1- (1H-indol-3-yl) propan-2-ylamino) -2-fluoro-2-methylpropan-1-ol

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3- [tert-Butyl (diphenyl) silyl] oxy-2-fluoro-N - [(7R) -2- (1H-indol-3-yl) -1-methylethyl] -2-methylpropan 1-amine (1,2 d, 2.4 mmol) was added to an oven dried 250 ml round bottom vessel and then THF (9.6 ml) and tetrabutylammonium fluoride hydrate (3 ml of a 1M solution in THF) were added. The reaction mixture was left stirring at room temperature until LCMS showed complete consumption of starting materials. The reaction mixture was quenched by the addition of water and extracted with 25% IPA in DCM, 5x100 ml. Then the combined organic matter was dried with MgSO ₄ , filtered and concentrated. Purification on a silica gel flash chromatography column (0-30% 2N NH ₃ in MeOH / DCM) afforded the title compound (332 mg, 53% yield). LCMS: 265.1 [M + H] +.

Step 7: 3 - ((1 R, 3R) -1 - (2,6-difluoro-4-iodophenyl) -3-methyl-3,4-dihydro-1 H-pyrido [3, 4-b] indole- 2 (9H) yl) -2-fluoro-2-methylpropan-1 -ol

Into a 100 ml round bottom vessel was added 2-fluoro-3 - [[(1R) -2- (1H-indol-3-yl) -1-methylethyl] amino] -2-methylpropan-1-ol (332 mg, 1 , 26 mmol), 2,6-difluoro-4-iodobenzaldehyde (370 mg, 1.1 eq., 1.38 mmol) and toluene (5.5 ml). The reaction was placed under nitrogen and acetic acid (2M) was added. The reaction mixture was then allowed to warm to 90 ° C for 48 h. The reaction was then quenched with saturated aqueous NaHCO ₃ and extracted vigorously with iPrOAc (5x100 ml). Then the organic phase was dried with MgSO ₄ , filtered and concentrated. Purification on a silica gel flash chromatography column (0-100% iPrOAc / heptanes) afforded the title compound (475 mg, 74% yield). LCMS: 515.1 [M + H] ⁺ .

Step 8: 3 - [(1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-1,3,4,9-tetrahydropyrido [3 , 4-b] indol-2-yl] -2-fluoro-2-methylpropan-1-ol (400 mg, 0.78 mmol), 2- [3- (fluoromethyl) azetidin-1-yl] ethanol (518 mg, 5 eq., 3.9 mmol), copper iodide (74 mg, 0.5 eq., 0.39 mmol) and potassium carbonate (644 mg, 6 eq., 4.7 mmol). The vial was covered and the mixture was placed under a nitrogen atmosphere. Then, butyronitrile (5.2 ml) was added and the mixture was degassed for 10 minutes. The reaction mixture was then heated to 135 ° C for 16 h, filtered through celite and purified by chiral reverse phase HPLC to give two diastereomers. 154 was the second eluted diastereomer (90 mg, 22% yield).

154: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.48 (s, 1H), 7.39 (dd, J = 7.4, 1.3 Hz, 1H), 7.17 (dd, J = 7.6, 1.2 Hz, 1H), 6.96 (dtd, J = 20.1, 7.2, 1.3 Hz, 2H), 6.72 - 6.55 (m, 2H) , 5.08 (s, 1H), 4.84 (t, J = 5.6 Hz, 1H), 4.56 (d, J = 6.2 Hz, 1H), 4.44 (d, J = 6.2 Hz, 1H), 3.92 (t, J = 5.4 Hz, 2H), 3.55 (q, J = 6.0, 5.4 Hz, 1H), 3.03 - 2, 83 (m, 4H), 2.72 (dt, J = 13.0, 5.6 Hz, 3H), 2.61 - 2.51 (m, 2H), 2.45 - 2.30 (m, 1H), 1.15-0.96 (m, 6H). The two protons are darkened under the peak of the water. Chiral HPLC: OX UPC2 column, isocratically 25% MeOH with 0.1% NH4OH for 2.5 min. Retention time 1.35 min. LCMS: 520.3 [M + H] ⁺ .

Example 155. (2R) -3 - [(1R, 3R) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetidin-1-yl] ethoxy] phenyl] -3methyl-1 , 3,4,9-tetrahydropyrido [3,4-b] indol-2-yl] -2-fluoro-2-methylpropan-1-ol 155.

According to the procedures of Example 154, 155 was the first eluted diastereomer (110 mg, 27% yield).

155: 1H NMR (400 MHz, DMSO): δ 10.52 (s, 1H), 7.42-7.34 (m, 1H), 7.21-7.14 (m, 1H), 6.96 (dtd, J = 20.9, 7.1, 1.2 Hz, 2H), 6.69 -6.58 (m, 2H), 5.12 (s, 1H), 4.81 (t, J = 5.8 Hz, 1H), 4.56 (d, J = 6.2 Hz, 1H), 4.44 (d, J = 6.2 Hz, 1H), 3.93 (t, J = 5 , 4 Hz, 2H), 3.46 (ddd, J = 18.2, 11.9, 5.7 Hz, 2H), 3.14 (ddd, J = 20.4, 11.9, 5.9 Hz, 2H), 3.03 - 2.78 (m, 4H), 2.78 - 2.64 (m, 3H), 2.58 - 2.51 (m, 2H), 2.47 - 2, 36 (m, 1H), 1.11 (d, J = 22.0 Hz, 3H), 1.04 (d, J = 6.5 Hz, 3H). The two protons are darkened under the peak of the water. Chiral CPLC: OX UPC2 column, isocratically 25% MeOH with 0.1% NH ₄ OH for 2.5 min. Retention time 0.55 min. LCMS: 520.2 [M + H] ⁺ .

Example 174. (1R, 3R) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetidin-1-yl] ethoxy] phenyl] -3-methyl-2 (2,2, 2-trifluoroethyl) -1,3,4,9-tetrahydropyrido [3,4-b] indole 174.

Step 1: (R) -1- (1H-indol-3-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine

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A mixture of (2R) -1- (1H-indol-3-yl) propan-2-amine (100 mg, 0.574 mmol), 2,2,2-trifluoroethyl trifluoromethanesulfonate (151 mg, 0.6313 mmol) and N, N- diisopropylethylamine α (371 mg, 2.87 mmol) in 1,4-dioxane (3.8261 ml) was heated at 50 ° C for 6 h. The reaction mixture was cooled to room temperature, diluted with water, extracted with EtOAc (2x). The combined organic matter was dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by silica gel flash chromatography (0-50% iPrOAc / heptane) to afford the title compound (89 mg, 60.5% yield) as a colorless oil.

¹ H NMR (chloroform-d): δ 8.10-7.92 (m, 1H), 7.62-7.56 (m, 1H), 7.33 (dt, J = 8.1, 0, 9 Hz, 1H), 7.23 - 7.16 (m, 1H), 7.15 - 7.08 (m, 1H), 7.02-6.98 (m, 1H), 3.21 - 3 09 (m, 3H), 2.83 (dd, J = 6.6, 0.8 Hz, 2H), 1.12 (d, J = 6.2 Hz, 3H). LCMS (electrospray ionization) m / z 257 [M + H +].

Step 2: (1 R, 3R) -1 - (2,6-difluoro-4-iodophenyl) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro- 1Hpyrido [3,4-b] indole

A mixture of (2R) -1- (1H-indol-3-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (54 mg, 0.211 mmol), 2,6 difluoro-4-iodobenzaldehyde (62 mg, 0.232 mmol) and acetic acid (110 mg, 1.84 mmol) in toluene (1 ml) was heated at 90 ° C for 5 The mixture was then concentrated. The precipitate was partitioned between EtOAc and saturated NaHCO ₃ solution. The aqueous layer was extracted with EtOAc (2x). The combined organic matter was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound as a white solid, which was used without purification. LCMS (electrospray ionization) m / z 507 [M + H +].

Stage 3: in a microwave oven the mixture (1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3methyl-2- (2,2,2-trifluoroethyl) -1,3,4 was poured, 9-tetrahydropyrido [3,4-b] indole (107 mg, 0.211 mmol), 2- [3 (fluoromethyl) azetidin-1-yl] ethanol (84 mg, 0.632 mmol), CuI (16 mg, 0.0843 mmol ) and K2CO3 (87 mg, 0.632 mmol) in butyronitrile (1.4 ml) was purged with nitrogen for 5 min and then sealed and heated at 135 ° C for 23 h. The mixture was filtered through celite, concentrated and purified by preparative HPLC with obtaining 174 (51 mg, 47% yield) as a yellow solid.

1H NMR (400 MHz, DMSO-de) δ 10.61 (s, 1H), 7.45-7.35 (m, 1H), 7.20 (dt, J = 8.0, 0.9 Hz, 1H), 7.05-6.90 (m, 2H), 6.71-6.59 (m, 2H), 5.20 (s, 1H), 4.50 (dd, J = 47.6, 6.2 Hz, 2H), 3.94 (t, J = 5.4 Hz, 2H), 3.57 - 3.35 (m, 2H), 3.31 - 3.22 (m, 2H), 2.97 (dt, J = 16.8, 7.9 Hz, 3H), 2.84 (ddd, J = 15.3, 4.9, 1.2 Hz, 1H), 2.77-2, 66 (m, 3H), 2.64-2.56 (m, 1H), 1.12 (d, J = 6.6Hz, 3H). LCMS (electrospray ionization) m / z 512 [M + H +].

Example 286. 3 - [(1R, 3R) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetidin-1-yl] ethoxy] phenyl] -3-methyl1,3,4 , 9-tetrahydropyrido [3,4-b] indol-2-yl] -2,2-difluoropropan-1-ol 286.

Step 1: 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-ol

TBDPSO / VF ^F

To a shaken solution of 2,2-difluoropropane-1,3-diol (200 mg, 1.78 mmol) in THF (4 ml) was added NaH (60% in mineral oil, 71 mg, 1.78 mmol) in an ice bath and the reaction mixture was shaken for 30 minutes. To the reaction mixture was added tert-butyl (chloro) diphenylsilane (TBDPSCI) (490 mg, 1.78 mmol) dropwise. The reaction mixture was then warmed to 20 ° C and shaking continued for 3 h. Water (10 ml) was slowly added to the reaction mixture and the resulting mixture was washed with EtOAc (10 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (20% petroleum ether in EtOAc) to give the title compound (450 mg, 1.28 mmol, 72% yield) as a light yellow oil.

1H NMR (400 MHz, CDCl3) δ 7.71 -7.64 (m, 4H), 7.44-7.36 (m, 6H), 3.96-3.84 (m, 4H), 1, 86 (s, 1H), 1.06 (s, 9H).

Step 2: 3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropyl trifluoromethanesulfonate

TBDPSO

OTf

F F

To a shaken solution of 3- [tert-butyl (diphenyl) silyl] oxy-2,2-difluoropropan-1-ol (from step 1, 400 mg, 1.14 mmol) and 2,6-lutidine (0.39 ml, 3.42 mmol) in DCM (8 ml) was added dropwise Tf ₂ O (0.38 ml, 2.28 mmol) in an ice bath. The reaction mixture was shaken at 20 ° C for 2 h. The reaction mixture was then slowly poured into ice water (20 ml) and the mixture was extracted with DCM (20 ml x 2). The combined organic layers were washed with 1N. HCl (20 ml), saturated NaHCO ₃ (20 ml) and brine. The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (10% petroleum ether in EtOAc) to give the desired product (500 mg, 1.04 mmol, 91%) as a light yellow oil.

1H NMR (400 MHz, CDCl ₃ ) δ 7.66 -7.64 (m, 4H), 7.47-7.41 (m, 6H), 4.76 (t, J = 7.6 Hz, 2H ), 3.89 (t, J = 7.6 Hz, 2H), 1.08 (s, 9H).

Step 3: (R) -N- (1- (1 H-indol-3-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-amine

A mixture of [3- [tert-butyl (diphenyl) silyl] oxy-2,2-difluoropropyl] trifluoromethanesulfonate (from step 2, 8.31 g, 17.22 mmol), diisopropylethylamine (DIPEA) (6.1 mL, 34, 44 mmol) and (2R) -1- (1H-indol-3yl) propan-2-amine (3 g, 17.22 mmol) in dioxane (60 ml) was shaken at 90 ° C for 12 h. After cooling to room temperature, the reaction mixture was diluted with water (100 ml) and washed with EtOAc (100 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (20% EtOAc in petroleum ether) to give the title compound (7.6 g, 87%) as a yellow oil. LCMS: 507.2 [M + H] ⁺ .

Step 4: (R) -3 - ((1- (1 H-indol-3-yl) propan-2-yl) amino) -2,2-difluoropropan-1-ol

To a shaken solution of (R) -N- (1- (1H-indol-3-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-amine (from step 3, 7.6 g, 15 mmol) in THF (100 ml) was added tetrabutylammonium fluoride (TBAF) (1.0 M in THF, 30 ml, 30 mmol). The reaction mixture was shaken at 25 ° C for 4 h and then diluted with water (200 ml) and was extracted with EtOAc (200 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (70% EtOAc in petroleum ether) to give the title compound (3.5 g, 87%) as a light yellow oil. LCMS: 268.9 [M + H] ⁺ .

Step 5: 3 - ((1 R, 3R) -1 - (2,6-difluoro-4-iodophenyl) -3-methyl-3,4-dihydro-1 H-pyrido [3,4-b] indole- 2 (9H) yl) -2,2-difluoropropane-1 -ol

A mixture of (R) -3 - ((1- (1H-indol-3-yl) propan-2-yl) amino) -2,2-difluoropropan-1-ol (from step 4, 2 g, 7.45 mmol ), HOAc (1.29 ml, 22.36 mmol) and 2,6-difluoro-4-iodobenzaldehyde (2 g, 7.45 mmol) in toluene (30 ml) was shaken at 90 ° C for 12 hours. cooling to room temperature, the reaction mixture was diluted with water (50 ml), washed with EtOAc (50 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (20% petroleum ether in EtOAc) to give the title compound (2.8 g, 73%) as a light yellow solid.

1H NMR (400 MHz, CDCl ₃ ) δ 7.53-7.49 (m, 2H), 7.30-7.22 (m, 3H), 7.18-7.13 (m, 2H), 5 , 25 (s, 1H), 3.72 - 3.68 (m, 3H), 3.24 - 3.06 (m, 3H), 2.85 - 2.75 (m, 1H), 2.70 - 2.66 (m, 1H), 1.18 (d, J = 6.8 Hz, 3H).

Step 6: a mixture of 3 - ((1R, 3R) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-3,4-dihydro-1H-pyrido [3,4-b] indole2 (9H ) -yl) -2,2-difluoropropan-1-ol (from step 5, 1.5 g, 2.89 mmol), 2- [3- (fluoromethyl) azetidin-1-yl] ethanol (1.93 g , 14.47 mmol), CuI (1.65 g, 8.68 mmol) and K2CO3 (1.2 g, 8.68 mmol) in n-PrCN (20 ml) were shaken in an N2 atmosphere at 135 ° C for 3 h. After cooling to room temperature, the reaction mixture was diluted with water (50 ml), washed with DCM (50 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The resulting residue was purified by reverse phase chromatography (acetonitrile 50-80% / 0.05% NH ₄ OH in water) to give 286 (170 mg, 11%) as a white solid.

¹ H NMR (400 MHz, CD3OD) δ 7.41 (d, J = 8.0 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.03 - 6.94 ( m, 2H), 6.54 (d, J = 11.2 Hz, 2H), 5.24 (s, 1H), 4.49 (dd, J = 47.6, 6.0 Hz, 2H), 4.00 - 3.98 (m, 2H), 3.83 - 3.72 (m, 1H), 3.63 - 3.45 (m, 4H), 3.22 - 3.13 (m, 3H ), 3.02-2.60 (m, 6H), 1.17 (d, J = 6.0Hz, 3H). LCMS: 524.1 [M + H] +.

Example 303 (1R, 3R) -2- (2,2-difluoroethyl) -1- [2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-yl] oxy-109 038835 phenyl] -3-methyl-1,3,4,9-tetrahydropyrido [3,4-b] indole 303.

Following the procedures of Example 305, 303 was prepared. LCMS: 494.2 [M + H] +.

Example 304. N- (3,5-difluoro-4 - ((1R, 3R) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro-1Hpyrido [3 , 4-b] indol-1-yl) phenyl) -1- (3-fluoropropyl) azetidin-3-amine 304.

Step 1: (R) -1- (1 H-indol-3-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine __Z _F F hnA ~ ^f

Η

To a solution of (2R) -1- (1H-indol-3-yl) propan-2-amine (10.0 g, 57.39 mmol) in 1,4-dioxane (100 ml) was added 2,2,2- trifluoroethyl trifluoromethanesulfonate (13.3 g, 57.39 mmol) and DIPEA (22.2 g, 172.18 mmol). The resulting mixture was shaken at 80 ° C for 15 h. The reaction mixture was concentrated and purified by column chromatography eluting with 0-30% EtOAc in hexanes to give the title compound (14 g, 95.2%) as a light yellow oils.

¹ H NMR (400 MHz, CDCl3) δ 8.02 (br.s., 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 8.0 Hz , 1H), 7.21 (t, J = 8.0 Hz, 1H), 7.16 - 7.09 (m, 1H), 7.05 (s, 1H), 3.24 - 3.11 ( m, 3H), 2.84 (d, J = 6.4 Hz, 2H), 1.14 (d, J = 6.4 Hz, 3H).

Stage 2: (1R, 3R) -1- (4-bromo-2,6-dufluorophenyl) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro-1Hpyrido [3, 4-b] indole

A mixture of (R) -1- (1H-indol-3-yl) -N- (2,2,2-trifluoroethyl) propan-2-amine (from stage 1, 14.0 g, 54.63 mmol), 4 α-bromo-2,6-difluorobenzaldehyde (11.5 g, 51.9 mmol) and acetic acid (6.25 ml, 109.26 mmol) in toluene (150 ml) was shaken at 90 ° C for 16 h. the mixture was cooled to 25 ° C, concentrated and purified by silica gel column chromatography (0-5% EtOAc in petroleum ether) to give the title compound and its cis isomer (24 g, 95.7% yield) (trans: cis is 4: 1) as a yellow solid.

1H NMR (400 MHz, CDCl ₃ ) δ 7.52 (d, J = 8.4 Hz, 1H), 7.24 (d, J = 8.0 Hz, 1H), 7.19-7.05 ( m, 4H), 5.69 (s, 0.2H), 5.31 (s, 0.8H), 3.64-3.50 (m, 1H), 3.45-3.17 (m, 1H), 3.10-3.06 (m, 1H), 2.98-2.81 (m, 1H), 2.78 2.59 (m, 1H), 1.41 (d, J = 6 , 4 Hz, 0.6H), 1.18 (d, J = 6.4 Hz, 2.4H).

Stage 3: tert-butyl-3 - ((3,5-difluoro-4 - ((1R, 3R) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9- tetrahydro-1Hpyrido [3,4-b] indol-1-yl) phenyl) amino) azetidine-1-carboxylate / - / ^F \ ^F QrCX-X

N AZ

A mixture of (1 R, 3R) -1 - (4-bromo-2,6-difluorophenyl) -3 -methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro-1H pyro [ 3,4-b] indole (trans: cis is 4: 1) (from step 2, 23.0 g, 50.08 mmol), Pd ₂ (dba) ₃ (4.59 g, 5.01 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (12.9 g, 75.12 mmol), Xantphos (5.8 g, 10.02 mmol) and Cs ₂ CO ₃ (48.9 g, 150.25 mmol ) in 1,4-dioxane (250 ml) was shaken at 115 ° C under N2 for 16 h. The reaction mixture was filtered through celite and the filtrate was concentrated and purified by column chromatography (0-30% EtOAc in petroleum ether) to give the title compound (25 g, 90.7% yield) (trans: cis 4: 1) as a light brown solid.

1H NMR (400 MHz, CDCl3) δ 7.56-7.37 (m, 1H), 7.24-7.19 (m, 1H), 7.15-7.06 (m, 2H), 6, 04 - 5.94 (m, 2H), 5.57 (s, 0.2H), 5.21 (s, 0.8H), 4.50 - 4.38 (m, 1H), 4.31 - 4.21 (m, 2H), 3.74-3.72 (m, 2H), 3.61-3.47 (m, 1H), 3.35-3.17 (m, 1H), 3, 10-3.07 (m, 1H), 3.02-2.78 (m, 1H), 2.77-2.55 (m, 1H), 1.44 (s, 9H), 1.39 ( d, J = 6.4Hz, 0.6H), 1.16 (d, J = 6.4Hz, 2.4H).

Step 4: N- (3,5-difluoro-4 - ((1R, 3R) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro-1H-pyrido [3,4b] indol-1-yl) phenyl) azetidine-3-amine

- 110 038835

To a solution of tert-butyl-3 - ((3,5-difluoro-4 - ((1R, 3R) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro1H -pyrido [3,4-b] indol-1-yl) phenyl) amino) azetidine-1-carboxylate (from step 3, 10.0 g, 18.16 mmol) (trans: cis is 4: 1) in 1 , 4-dioxane (120 ml) sulfuric acid (4.87 ml, 90.82 mmol) was added at 0 ° C. The reaction mixture was shaken at 0 ° C for 0.5 h. The reaction mixture was poured into saturated aqueous NaHCO ₃ (250 ml) and the mixture was extracted with EtOAc (200 ml x 2). The combined organic layers were dried over Na ₂ SO ₄ and concentrated to give the title compound (8 g, 97.8% yield) (trans: cis 4: 1) as a yellow solid. The crude compound was used directly in the next step.

Step 5: to a mixture of n- (3,5-difluoro-4 - ((1R, 3R) -3-methyl-2- (2,2,2-trifluoroethyl) -2,3,4,9-tetrahydro-1Hpyrido [3,4-b] indol-1-yl) phenyl) azetidin-3-amine (from step 4, trans: cis is 4: 1, 8.0 g, 17.76 mmol) and DIPEA (8.83 ml , 53.28 mmol) in N, N-dimethylformamide (DMF) (80 ml) was added dropwise 1-fluoro-3-iodopropane (3.34 g, 17.76 mmol). The reaction mixture was shaken at 20 ° C for 16 h. The reaction mixture was diluted with EtOAc (400 ml), washed with brine (200 ml x 5). The combined organic layers were dried over Na ₂ SO ₄ , filtered, concentrated and purified by column chromatography (10-40% EtOAc in DCM) to give the desired product (7 g, 77.2% yield) as a brown solid. This product was combined with another batch (12.3 g total) and purified by preparative HPLC (Phenomenex Synergi Max-RP 250x80 mm, 10 μm, acetonitrile 50-80 / 10 mM NH4HCO3 in water) to give 10 g of product (trans: cis is 4: 1, not separable by HPLC) as a white solid. This product (trans: cis is 4: 1) was then purified further by CPLC (AD (250 mm x 30 mm, 10 μm) base-EtOH 40%) to give pure 304 (5.9 g, 59% yield) as white solid.

1H NMR (400 MHz, CD3OD) δ 7.40 (d, J = 7.6 Hz, 1H), 7.20 (d, J = 7.6 Hz, 1H), 7.05 - 6.91 (m , 2H), 6.09 (d, J = 12 Hz, 2H), 5.22 (s, 1H), 4.58-4.35 (m, 2H), 4.07-4.02 (m, 1H), 3.77 (t, J = 7.6 Hz, 2H), 3.62-3.50 (m, 1H), 3.39-3.32 (m, 1H), 3.06-2 , 90 (m, 4H), 2.66-2.55 (m, 3H), 1.87-1.66 (m, 2H), 1.17 (d, J = 6.4Hz, 3H).

Example 305 (1R, 3R) -2- (2,2-difluoroethyl) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetidin-1-yl] ethoxy] phenyl] - 3-methyl-1,3,4,9-tetrahydropyrido [3, 4-b] indole 305.

Step 1: (R) -N- (2,2-difluoroethyl) -1- (1H-indol-3-yl) pronan-2-amine

A mixture of (2R) -1- (1H-indol-3-yl) propan-2-amine (4.2 g, 24.1 mmol), 2,2-difluoroethyl trifluoromethanesulfonate (5.16 g, 24.1 mmol) and diisopropylamine (8.41 ml, 48.2 mmol) was heated to 80 ° C for 3 h. The reaction was cooled to room temperature, diluted with iPrOAc (150 ml) and washed with water, brine, dried over sodium sulfate, filtered and concentrated ... The crude product was purified on a silica gel flash chromatography column eluting with 05% MeOH / DCM to give the title compound (5.6 g, 97% yield).

1H NMR (400 MHz, chloroform-d) δ 8.03 (s, 1H), 7.63-7.54 (m, 1H), 7.36 (dt, J = 8.1, 0.9 Hz, 1H), 7.27-7.17 (m, 1H), 7.12 (ddd, J = 8.0, 7.1, 1.1 Hz, 1H), 5.78 (tdd, J = 56, 6, 4.7, 4.1 Hz, 1H), 3.11 -2.76 (m, 5H), 1.12 (d, J = 6.2 Hz, 3H); LCMS: 239.15 [M + H] +.

Stage 2: (1R, 3R) -2- (2,2-difluoroethyl) -1- (2,6-difluoro-4-iodo-phenyl) -3-methyl-1,3,4,9-tetrahydropyrido [3 , 4-b] indole

To a solution of (R) -N- (2,2-difluoroethyl) -1- (1H-indol-3-yl) npropan-2-amine (5.0 g, 21 mmol) and 2,6 difluoro-4-iodobenzaldehyde ( 5.2 g, 19 mmol) in toluene (70 ml), acetic acid (2.4 ml) was added and the mixture was heated at 90 ° C for 20 h under nitrogen atmosphere. The reaction mixture was cooled and concentrated. The precipitate was dissolved in iPrOAc and washed with saturated sodium bicarbonate solution, water,

- 111 038835 brine, dried over sodium sulfate and concentrated. Purification by flash chromatography (silica gel, 0-15% iPrOAc / heptanes) gave the title compound (7.8 g, 76%) as a 3: 1 mixture of trans: cis isomers. LCMS: 489.0 [M + H] +. The mixture was used in the next step as such.

Stage 3: a mixture of (1R, 3R) -2- (2,2-difluoroethyl) -1- (2,6-difluoro-4-iodophenyl) -3-methyl-1,3,4,9-tetrahydropyrido [3, 4-b] indole (8.0 g, 16.4 mmol), 2- [3- (fluoromethyl) azetidin-1-yl] ethanol (2.62 g, 19.7 mmol), copper iodide (0.94 g, 4.9 mmol), potassium carbonate (4.5 g, 32.8 mmol) and butyronitrile (33 ml) were degassed for 5 min and then heated to 140 ° C overnight. The reaction mixture was filtered through celite eluting with iPrOAc. The filtrate was concentrated and purified by reverse phase HPLC and the cis: trans isomers were separated by chiral HPLC to give 305 (3.77 g, 44% yield).

¹ H NMR (400 MHz, DMSOC) δ 10.59 (s, 1H), 7.40 (dd, J = 7.9, 1.3 Hz, 1H), 7.22 - 7.14 (m, 1H ), 7.04 6.88 (m, 2H), 6.66 (d, J = 11.1 Hz, 2H), 6.05-5.61 (m, 1H), 5.17 (d, J = 1.7 Hz, 1H), 4.50 (dd, J = 47.6, 6.2 Hz, 2H), 3.94 (t, J = 5.3 Hz, 2H), 3.41 - 3 , 32 (m, 2H), 3.15 - 2.90 (m, 3H), 2.90 - 2.52 (m, 7H), 1.09 (d, J = 6.5 Hz, 3H). LCMS: 494.2 [M + H] ⁺ .

Example 306 (1S, 3R) -2- (2,2-difluoroethyl) -1- [2,6-difluoro-4- [2- [3- (fluoromethyl) azetidin-1-yl] ethoxy] phenyl] - 3-methyl-1,3,4,9-tetrahydropyrido [3,4-b] indole 306.

Following the procedures of Example 305, 306 was prepared. LCMS: 494.2 [M + H] ⁺ .

Example 340. 3 - [(1R, 3R) -1- [2,6-difluoro-4 - [[1- (3-fluoropropyl) αzetidin-3-yl] amino] phenyl] -3-methyl1,3,4 , 9-tetrahydropyrido [3,4-b] indol-2-yl] -2,2-difluoropropan-1-ol 340.

Step 1: (R) -N- (1- (1H-indol-3-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-amine

A mixture of (2H) -1 - (1 N-indol-3-yl) propan-2-amine (29 g, 166.44 mmol), [3- [tert-butyl (diphenyl) silyl] oxy-2,2 -difluoropropyl] trifluoromethanesulfonate (80.31 g, 166.44 mmol) and DIPEA (55.01 ml, 332.87 mmol) in 1,4-dioxane (600 ml) was shaken at 90 ° C for 12 hours. The reaction mixture was diluted in water (600 ml) and was extracted with EtOAc (600 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (40% EtOAc in petroleum ether) to give the title compound (69 g, 82%) as a light yellow oil.

Ή NMR (400 MHz, CDCl ₃ ) δ 7.88 (s, 1H), 7.66 (d, J = 7.2 Hz, 4H), 7.60 (d, J = 8.0 Hz, 1H) , 7.48-7.33 (m, 7H), 7.22-7.08 (m, 2H), 7.01 (s, 1H), 3.86 -3.79 (m, 2H), 3 , 23 - 3.09 (m, 3H), 2.86 - 2.80 (m, 2H), 1.13 (d, J = 6.4 Hz, 3H), 1.05 (s, 9H). MS: [M + H] + 507.1.

Stage 2: (R) -3 - ((1- (1 N-indol-3-yl) propan-2-yl) amino) -2,2-difluoropropane-1 -o l

To a shaken solution of (R) -N- (1- (1H-indol-3-yl) propan-2-yl) -3 - ((tert-butyldiphenylsilyl) oxy) -2,2-difluoropropan-1-amine (from step 1, 69 g, 136.18 mmol) in THF (690 ml) was added 1M TBAF (272.35 ml, 272.35 mmol) in THF. The mixture was shaken at 25 ° C for 4 h. The reaction mixture was diluted with water (800 ml) and was extracted with EtOAc (800 ml x 3). The combined organic layers were concentrated and the crude residue was purified by silica gel column chromatography (50% EtOAc in petroleum ether) to give the title compound (29 g, 79%) as a light yellow oil.

Stage 3: 3 - ((1R, 3R) -1 - (4-bromo-2,6-difluorophenyl) -3-methyl-3,4-dihydro-1 H-pyrido [3, 4-b] indole-2 (9H) yl) -2,2-difluoropropane-1 -ol

A mixture of (R) -3 - ((1- (1H-indol-3-yl) propan-2-yl) amino) -2,2-difluoropropan-1-ol (from step 2, 20 g, 74.54 mmol ), acetic acid (12.91 ml, 223.63 mmol) and 4-bromo-2,6-difluorobenzaldehyde (16.47 g, 74.54 mmol) in toluene (400 ml) were shaken at 90 ° C for 12 h. The reaction mixture was diluted with water (500 ml) and was extracted with EtOAc (500 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (20% EtOAc in petroleum ether) to give the title compound (24.8 g, 71%, trans: cis 20/1) as a light yellow solid. MS: [M + H] + 470.9.

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Step 4: tert-butyl-3 - ((4 - ((1R, 3R) -2- (2,2-difluoro-3-hydroxypropyl) -3-methyl-2,3,4,9-tetrahydro-1Hpyrido [ 3,4-b] indol-1-yl) -3,5-difluorophenyl) amino) azetidine-1-carboxylate

A mixture of 3 - ((1R, 3R) -1- (4-bromo-2,6-difluorophenyl) -3-methyl-3,4-dihydro-1H-pyrido [3,4-b] indole-2 (9H) yl) -2,2-difluoropropan-1-ol (from stage 3, 24.8 g, 52.62 mmol), Pd ₂ (dba) ₃ (4.82 g, 5.26 mmol), Xantphos (6, 09 g, 10.52 mmol), Cs ₂ CO ₃ (51.44 g, 157.86 mmol) and tert-butyl 3-aminoazetidine-1-carboxylate (13.59 g, 78.93 mmol) in 1, 4-dioxane (300 ml) was shaken at 110 ° C for 3 h under N ₂ . The reaction mixture was cooled to 25 ° C and diluted with water (500 ml), extracted with EtOAc (500 ml x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (20% petroleum ether in EtOAc) to give the title compound (20.5 g, 69%, trans: cis 20/1) as a yellow solid. MS: [M + H] + 563.0.

Step 5: 3 - ((1 R, 3R) -1 - (4- (azetidin-3-ylamino) -2,6-difluorophenyl) -3-methyl-3,4-dihydro-1 H-pyrido [3, 4-b] indol-2 (9H) -yl) -2,2-difluoropropan-1-ol

To a solution of tert-butyl-3 - ((4 - ((1R, 3R) -2- (2,2-difluoro-3-hydroxypropyl) -3-methyl-2,3,4,9-tetrahydro1H-pyrido [3 , 4-b] indol-1-yl) -3,5-dufluorophenyl) amino) azetidine-1-carboxylate (from step 4, 20.5 g, 36.44 mmol) in 1,4-dioxane (194 ml) sulfuric acid (19.42 mL, 364.38 mmol) was added dropwise in an ice bath. The reaction mixture was shaken at 25 ° C for 0.5 h. Then the reaction mixture was poured into saturated aqueous NaHCO ₃ solution (800 ml) and was extracted with EtOAc (600 ml x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated to give the title compound (18 g, crude, trans: cis 20/1) as a yellow solid. The crude precipitate was used directly in the next step. MS: [M + H] + 463.0.

Stage 6: a mixture of 3 - ((1R, 3R) -1- (4- (azetidin-3-ylamino) -2,6-difluorophenyl) -3-methyl-3,4-dihydro-1Hnirudo [3,4-b ] indol-2 (9H) -yl) -2,2-difluoropropan-1-ol (from step 5, 18 g, 38.92 mmol), DIPEA (19.3 ml, 116.76 mmol) and 1-fluoro -3-iodopropane (7.32 g, 38.92 mmol) in DMF (180 ml) was shaken at 25 ° C for 12 h. The reaction mixture was diluted with EtOAc (500 ml) and washed with brine (500 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (10% MeOH in DCM) to give the desired product (7.1 g, 85% pure) as a yellow oil. The resulting residue was further purified by reverse phase chromatography (acetonitrile 40-75 / 0.05% NH4OH in water) and chiral HPLC (AD 250 mm x 50 mm, 10 μm; supercritical CO ₂ / EtOH (0.1% NH ₃ H ₂ O) is 40/40 at 200 ml / min), giving 340 (2.85 g, 14%) as a light yellow solid.

1H NMR (400 MHz, CD ₃ OD) δ 7.39 (d, J = 7.2 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.01 - 6.93 (m, 2H), 6.11 (d, J = 12.0 Hz, 2H), 5.16 (s, 1H), 4.52-4.38 (m, 2H), 4.05-4, 03 (m, 1H), 3.80-3.74 (m, 3H), 3.63-3.42 (m, 2H), 3.203.10 (m, 1H), 2.96-292 ( m, 3H), 2.82 - 2.71 (m, 1H), 2.64-2.58 (m, 3H), 1.81 - 1.68 (m, 2H), 1.14 (d, J = 6.4 Hz, 3H); MS: [M + H] + 523.2.

Example 365.3 - ((1R, 3R) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro-3methyl-1 , 3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2,2-difluoropropan-1-ol 365.

Step 1: ([3- (tert-butyldiphenylsilanyloxy) -2,2-dufluoropropyl] - [2- (5-fluoro-1H-indol-3-yl) -1methylethyl] amine

A mixture of [3- [tert-butyl (diphenyl) silyl] oxy-2,2-difluoropropyl] trifluoromethanesulfonate (from example

- 113 038835

286, step 2, 43.22 g, 89.6 mmol), DIPEA (19.5 ml, 112.0 mmol) and 2- (5-fluoro-III-indol-3-yl) -1methylethylamine (CAS No. 712 -08-3, 14.7 g, 74.7 mmol) in dioxane (140 ml) was shaken at 90 ° C for 3 h. The mixture was cooled to room temperature, diluted with EtOAc and washed with water (x2) and brine solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel column chromatography (mobile phase: DCM) to give the title compound (32.8 g, 96%) as a yellow oil.

1H NMR (400 MHz, CDCl ₃ ): δ 7.89 (s, 1H), 7.69-7.61 (m, 4H), 7.48-7.34 (m, 6H), 7.26- 7.19 (m, 2H), 7.03 - 7.02 (m, 1H), 6.93 (dt, J = 2.5, 9.0 Hz, 1H), 3.88 - 3.78 ( m, 2H), 3.22 - 3.03 (m, 3H), 2.84 - 2.70 (m, 2H), 1.11 (d, J = 6.2 Hz, 3H), 1.04 (s, 9H). LCMS: 525.3 [M + H] +.

Step 2: 2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -1- (2,6-difluoro-4-iodophenyl) -6fluoro-3-methyl-2,3,4,9-tetrahydro -1H-β-carboline

To a solution of [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] - [2- (5-fluoro-1H-indol-3-yl) -1methylethyl] amine (32.8 g, 62.5 mmol) in toluene (65 ml) was added 4-iodo-2,6-difluorobenzaldehyde (20.1 g, 75.0 mmol) and acetic acid (7.2 ml, 125.0 mmol). Upon completion of the addition, the reaction mixture was shaken at 90 ° C for 14 h. The mixture was allowed to cool to room temperature, diluted with EtOAc, washed with saturated aqueous NaHCO ₃ (x3) and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by chromatography on silica gel (mobile phase: toluene in cyclohexane, gradient 10-50%) to give the title compound (34.9 g, 72%) as a white foam with a yellowish tint.

1H NMR (400 MHz, CDCl3): δ 7.65-7.60 (m, 4H), 7.46-7.33 (m, 7H), 7.21-7.08 (m, 4H), 6 , 90 - 6.84 (m, 1H), 5.27 (s, 1H), 3.99 - 3.88 (m, 1H), 3.65 - 3.54 (m, 2H), 3.33 - 3.20 (m, 1H), 2.93 (ddd, J = 1.4, 4.9, 15.2 Hz, 1H), 2.81 - 2.69 (m, 1H), 2.56 - 2.51 (m, 1H), 1.14 (d, J = 6.6 Hz, 3H), 1.05 (s, 9H). LCMS: 775.2 [M + H] +.

Step 3: 3- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -6-fluoro-3-methyl-2,3,4,9tetrahydro-1 H-β-carboline- 1 -yl} -3,5-difluorophenylamino-azetidine-1-carboxylic acid tert-butyl ester

A mixture of 2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -1- (2,6-difluoro-4-iodophenyl) -6fluoro-3-methyl-2,3,4,9-tetrahydro-1H -β-carboline (30.8 g, 39.8 mmol), XantPhos (4.60 g, 7.9 mmol), Pd ₂ (dba) ₃ (3.64 g, 4.0 mmol), Cs ₂ CO ₃ (25.9 g, 79.4 mmol) and tert-butyl 3-aminoazetidine-1-carboxylate (10.3 g, 59.6 mmol) in 1,4-dioxane (192 ml) was shaken at 115 ° C in a sealed vessel under argon atmosphere for 1.5 hours. The reaction mixture was allowed to cool to room temperature, the remaining solid was removed by filtration through a Celite® pad and the filtrate was concentrated. The resulting residue was purified by flash chromatography on silica gel (mobile phase: EtOAc in DCM, gradient 0-5%) to afford the title compound (27.9 g, 76%) as a beige foam.

1H NMR (400 MHz, CDCl3): δ 7.67-7.58 (m, 4H), 7.48-7.32 (m, 6H), 7.15-7.06 (m, 2H), 6 , 88 - 6.80 (m, 1H), 5.88 - 5.80 (m, 2H), 5.15 (s, 1H), 4.26 - 4.09 (m, 2H), 4.03 - 3.91 (m, 2H), 3.69 - 3.50 (m, 3H), 3.26 3.14 (m, 1H), 2.95 - 2.90 (m, 1H), 2, 83 - 2.70 (m, 1H), 2.50 (dd, J = 3.3, 15.1 Hz, 1H), 1.44 (s, 9H), 1.13 (d, J = 6, 5 Hz, 3H), 1.04 (s, 9H). LCMS: 819.4 [M + H] +.

Step 4: azetidin-3-yl- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -6-fluoro-3methyl-2,3,4,9-tetrahydro-1H-β -carbolin-1-yl} -3,5-difluorophenyl) amine

- 114 038835

A previously mixed and ice-cooled solution of concentrated sulfuric acid (9.1 ml, 170.2 mmol) in dioxane (100 ml) was slowly added to a solution of 3- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2, 2-difluoropropyl] -6-fluoro-3-methyl-2,3,4,9-tetrahydro-III-v-carbolin-1-yl} -3,5-difluorophenylamino) azetidine-1-carboxylic acid tert-butyl ester (27.9 g, 34.0 mmol) in dioxane (275 ml) under nitrogen at room temperature. After the addition was complete, the reaction mixture was left for 1 hour at room temperature. Added EtOAc and water and the pH of the aqueous phase was brought to pH 9 by adding solid Na ₂ CO ₃ . The organic layer was separated, washed with brine (x3), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound (mixture of diastereoisomers (R, R) and (S, S)) as a pale orange foam ( 26.6 g, approximate amount). LCMS: 719.4 [M + H] +.

Step 5: (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -6-fluoro-3-methyl-2,3,4,9tetrahydro-1H-β-carbolin-1-yl } -3,5-difluorophenyl) - [1- (3-fluoropropyl) azetidin-3-yl] amine

F

The title compound was obtained from azetidin-3-yl- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2,2difluoropropyl] -6-fluoro-3-methyl-2,3,4,9-tetrahydro-1H- β-carbolin-1-yl} -3,5-difluorophenyl) amine (26.6 g, 34.0 mmol) and 1-iodo-3-fluoropropane (9.60 g, 51.1 mmol; CAS No. 462- 40-8) following the procedure outlined for the preparation of Example 101. The crude product was purified by silica gel chromatography (mobile phase: dichloromethane / methanol, gradient 0 to 5%) to afford the title compound as a pale brown foam (14, 9 g, 56%).

1H NMR (400 MHz, CDCl ₃ ): δ 7.69 - 7.56 (m, 4H), 7.50 (s, 1H), 7.47-7.30 (m, 6H), 7.16 - 7.01 (m, 2H), 6.87 - 6.81 (m, 1H), 5.92 - 5.78 (m, 2H), 5.14 (s, 1H), 4.54 (t, J = 6.0 Hz, 1H), 4.42 (t, J = 6.0 Hz, 1H), 4.18 (d, J = 7.0 Hz, 1H), 4.06 - 3.86 ( m, 2H), 3.74-3.47 (m, 4H), 3.30-3.10 (m, 1H), 3.00-2.70 (m, 3H), 2.56 (t, J = 7.2 Hz, 2H), 2.53 - 2.45 (m, 1H), 1.85-1.50 (m, 3H), 1.12 (d, J = 6.5 Hz, 3H ), 1.04 (s, 9H). LCMS: 779.4 [M + H] +.

Step 6: 3- (1- {2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-ylamino] phenyl} -6-fluoro-3-methyl-1,3,4,9tetrahydro- β-carbolin-2-yl) -2,2-difluoropropan-1-ol

F

To a mixture (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2,2-difluoropropyl] -6-fluoro-3-methyl-2,3,4,9tetrahydro-1H-β-carbolin-1-yl} -3,5-difluorophenyl) - [1- (3-fluoropropyl) azetidin-3-ul] amine (12.1 g, 15.5 mmol) in THF (150 ml) in an argon atmosphere, a 1M solution of TBAF in THF ( 23.3 ml) and the reaction mixture was shaken at room temperature for 5 h. The reaction mixture was diluted with EtOAc and washed with water (x4). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (mobile phase: 2M ammonium in methanol / MTBE (methyl tert-butyl ether), gradient 0.5 to 5%) to give a mixture of (R, R) - and (S, S) -isomers of 3- (1- {2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-ylamino] phenyl} -6-fluoro-3-methyl-1,3,4,9tetrahydro-v -carbolin-2-yl) -2,2-difluoropropan-1-ol. The pair of diastereoisomers were separated by chiral HPLC (ChiralPak IB, 15% EtOH in heptane plus 0.1% diethylamine). 365 was the second peak isolated by chiral HPLC: (1.90 g, 23%). rt (retention time) of peak 2 is 15 min.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.46 (s, 1H), 7.16-7.08 (m, 2H), 6.86 (dt, J = 2.5, 9.0 Hz , 1H), 6.08 - 6.00 (m, 2H), 5.09 (s, 1H), 4.54 (t, J = 5.9 Hz, 1H), 4.43 (t, J = 5.9 Hz, 1H), 4.38 (d, J = 6.7 Hz, 1H), 4.07 - 3.97 (m, 1H), 3.80 - 3.56 (m, 6H), 3.28 - 3.16 (m, 1H), 3.11 - 3.02 (m, 1H), 2.97 - 2.82 (m, 3H), 2.64 - 2.55 (m, 3H ), 1.82 1.67 (m, 2H), 1.16 (d, J = 6.5 Hz, 3H). LCMS: 541.4 [M + H] ⁺ .

Example 366.3 - ((1S, 3S) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro-3methyl-1 , 3,4,9-tetrahydro-2H-pyrido [3,4-b] undol-2-yl) -2,2-difluoropropan-1-ol 366.

Following the procedures of Example 365, the first peak isolated by chiral HPLC was 366: (1.95 g, 24%). rt of peak 1 is 12 min.

- 115 038835

1H NMR (400 MHz, CDCl ₃ ): δ 7.46 (s, 1H), 7.16-7.08 (m, 2H), 6.86 (dt, J = 2.5, 9.0 Hz, 1H), 6.08 - 6.00 (m, 2H), 5.09 (s, 1H), 4.54 (t, J = 5.9 Hz, 1H), 4.43 (t, J = 5 , 9 Hz, 1H), 4.38 (d, J = 6.7 Hz, 1H), 4.07 - 3.97 (m, 1H),

3.80 - 3.56 (m, 6H), 3.28 - 3.16 (m, 1H), 3.11 - 3.02 (m, 1H), 2.97 - 2.82 (m, 3H ), 2.64 - 2.55 (m, 3H), 1.82 1.67 (m, 2H), 1.16 (d, J = 6.5 Hz, 3H). LCMS: 541.4 [M + H] +.

Example 368.3 - ((1 R, 3R) -1 - (2,6-difluoro-4 - ((1 - (3-fluoropropyl) azetidin-3-yl) oxy) phenyl) -6-fluoro-3 methyl -1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 368.

Step 1: [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] - [2- (5-fluoro-1H-indol-3-yl) -1methylethyl] amine

To a solution of 2- (5-fluoro-1H-indol-3-yl) -1-methylethylamine (CAS No. 712-08-3, 3.61 g, 18.7 mmol)) and DIPEA (4.9 ml, 28 , 1 mmol) in dioxane (43 ml) under argon was added trifluoromethanesulfonic acid 3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl ether, intermediate XX (3.09 g, 9.48 mmol). The resulting mixture was shaken at 90 ° C for 6 h. The reaction mixture was partitioned between EtOAc and water. The organic phase was separated and further washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (mobile phase: dichloromethane / methanol, gradient 0 to 5%) to give a mixture of diastereoisomers of the title compound as a yellow oil (8.0 g, 82%).

1H NMR (300 MHz, CDCl ₃ ): δ 7.82 (br. S, 1H), 7.68-7.63 (m, 4H), 7.52-7.38 (m, 6H), 7, 25 - 7.18 (m, 2H), 7.02 - 6.98 (m, 1H), 6.92 (dt, J = 2.4, 9.1 Hz, 1H), 3.82-3, 57 (m, 5H), 3.02-2.65 (m, 6H), 1.33 (d, J = 22.0 Hz, 3H), 1.1 -1.0 (m, 9H); LCMS: 521.3 [M + H] ⁺ .

Step 2: 3- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] -6-fluoro-3-methyl-2,3,4,9tetrahydro-1H-β-carboline -1-yl} -3,5-difluorophenoxy) azetidine-1-carboxylic acid tert-butyl ester

The title compound was prepared from [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] - [2 (5-fluoro-1H-indol-3-ul) -1-methylethyl] amine, intermediate 1a (8 g , 15.3 mmol), and 3- (3,5 difluoro-4-formylphenoxy) azetidine-1-carboxylic acid tert-butyl ester 101c (5.6 g, 18.1 mmol), following the procedure outlined for the preparation of the intermediate 101e. The crude product was purified by silica gel chromatography (mobile phase: cyclohexane / ethyl acetate, gradient 0 to 20%) to provide a mixture of diastereoisomers of the title compound as a white foam (8.0 g, 64%).

1H NMR (300 MHz, CDCl ₃ ): δ 7.65-7.54 (m, 4H), 7.47-7.30 (m, 7H), 7.17-7.08 (m, 2H), 6.84 (dt, J = 2.4, 9.0 Hz, 1H), 6.23-6.07 (m, 2H), 5.16 (s, 1H), 4.71-4.63 ( m, 1H), 4.29 - 4.18 (m, 2H), 3.99 - 3.88 (m, 2H), 3.79 (dd, J = 11.5, 16.8 Hz, 1H) , 3.64 - 3.54 (m, 1H), 3.50 - 3.29 (m, 1H), 3.10 - 2.83 (m, 2H), 2.69 - 2.39 (m, 2H), 1.54 (s, 3H), 1.46-1.40 (m, 9H), 1.29-0.98 (m, 12H); LCMS: 816.5 [M + H] ⁺ .

Step 3: 1- [4- (azetidin-3-yloxy) -2,6-difluorophenyl] -2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro2-methylpropyl] -6-fluoro-3-methyl-2 , 3,4,9-tetrahydro-1H-β-carboline

To a mixture of 3- (4- {2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] -6-fluoro-3-methyl2,3,4,9-tetrahydro-1 H-в-carboline -1 -yl} -3,5-difluorophenoxy) azetidine-1-carboxylic acid tert-116 038835 butyl ester, intermediate 2a (8.0 g, 9.80 mmol), in dioxane (80 ml) under argon at At 0 ° C, a solution of concentrated sulfuric acid (2.62 ml, 49.0 mmol) in dioxane (27 ml) was added dropwise, and the mixture, protected from light, was allowed to warm to RT and shaken for 3.5 h. The reaction mixture diluted with EtOAc and saturated NaHCO ₃ solution, shaken for 10 min and the layers were separated. The organic layer was then washed with saturated NaHCO ₃ solution, brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a mixture of diastereoisomers of the title compound as a pale yellow foam (7.05 g, approximate amount).

1H NMR (300 MHz, CDCl3): δ 7.66-7.54 (m, 4H), 7.47-7.30 (m, 7H), 7.17-7.06 (m, 2H), 6 , 84 (dt, J = 2.4, 9.0 Hz, 1H), 6.25-6.09 (m, 2H), 5.16 (s, 1H), 4.86-4.76 (m , 1H), 3.94-3.65 (m, 3H), 3.63-3.54 (m, 1H), 3.49 3.24 (m, 1H), 3.11-2.83 ( m, 2H), 2.69-2.39 (m, 2H), 1.54 (s, 3H), 1.27-1.12 (m, 3H), 1.11-0.98 (m, 12H); LCMS: 716.4 [M + H] +.

Step 4: 2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] -1- {2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-yloxy] phenyl} -6-fluoro-3-methyl-2,3,4,9-tetrahydro-1H-β-carboline

F

The title compound was prepared from 1- [4- (azetidin-3-yloxy) -2,6-difluorophenyl] -2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] -6-fluoro-3- methyl 2,3,4,9-tetrahydro-1H-β-carboline, intermediate 3a (7.05 g, 9.84 mmol), and 1-iodo-3-fluoropropane (2.77 g, 14.7 mmol; CAS # 462-408) following the procedure outlined for Example 101. The crude product was purified by silica gel chromatography (mobile phase: dichloromethane / methanol, gradient 0 to 3%) to afford the title compound as a white foam (5 , 7 g, 75%). LCMS: 776.4 [M + H] ⁺ .

Step 5: racemic 3- (1- {2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-yloxy] phenyl} -6-fluoro-3methyl-1,3,4,9-tetrahydro -v-carbolin-2-yl) -2-fluoro-2-methylpropan-1 -ol

F

To a mixture of 2- [3- (tert-butyldiphenylsilanyloxy) -2-fluoro-2-methylpropyl] -1- {2,6-difluoro-4- [1- (3-fluoropropyl) azetidin-3-yloxy] phenyl} - 6-fluoro-3-methyl-2,3,4,9-tetrahydro-1H-β-carboline, intermediate 4a (5.17 g, 6.66 mmol), 1M a solution of TBAF in THF (10 ml), the reaction mixture was shaken at RT for 24 h. The reaction mixture was poured into a mixture of water and brine and was extracted with EtOAc. The aqueous layer was then extracted with EtOAc and the combined organic layers were further washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (mobile phase: dichloromethane / methanol, gradient 0 to 6%) to give two pairs of diastereoisomers (diastereoisomers 1 and diastereoisomers 2). A pair of diastereoisomers 1 was further purified by chiral HPLC (ChiralPak IC, 25% IPA in heptane, 0.1% diethylamine). The first highlighted peak (rt is 8.2 min) is 368, isolated as a white solid (467 mg, 13%).

1H NMR (400 MHz, CDCl3): 7.32 (br s., 1H), 7.17-7.09 (m, 2H), 6.89-6.83 (m, 1H), 6.38 - 6.33 (m, 2H), 5.03 (s, 1H), 4.76 - 4.69 (m, 1H), 4.55 (t, 1H, J = 5.9 Hz), 4.47 - 4.41 (m, 2H), 4.00 (t, 1H, J = 4.9 Hz), 3.83 - 3.76 (m, 2H), 3.60 (q, 1H, J = 10 , 3 Hz), 3.46 - 3.34 (m, 1H), 3.23 - 3.09 (m, 4H), 2.67 - 2.57 (m, 4H), 1.84 - 1, 69 (m, 2H), 1.14-1.08 (m, 6H); LCMS: 538.3 [M + H] ⁺ .

Example 369.3 - ((1S, 3S) -1- (2,6-difluoro-4 - ((1- (3-fluoro-propyl) azetidin-3-yl) oxy) phenyl) -6-fluoro-3methyl-1 , 3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylnron-1-ol 369.

Following the procedures of Example 368, the second peak, isolated by chiral HPLC (rt = 15.5 min) is 369, was isolated as a white solid (480 mg, 13.5%).

Example 370.3 - ((1R, 3R) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) oxy) phenyl) -6-fluoro-3methyl-1 , 3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 370.

Following the procedures of Example 368, a pair of diastereoisomers 2 were purified by chiral HPLC

117 038835 (ChiralPak IC, 35% IPA in heptane, 0.1% diethylamine). The first isolated peak was further purified by chiral HPLC (ChiralPak IA, 25% IPA in heptane, 0.1% diethylamine): the first isolated peak (rt is 8.5 min) is 370, isolated as a white solid (165 mg, 5 %).

1H NMR (400 MHz, CDCl ₃ ): 7.53 (br. S, 1H), 7.16 - 7.12 (m, 2H), 6.90 - 6.84 (m, 1H), 6.33 - 6.28 (m, 2H), 5.35 (s, 1H), 4.76 - 4.68 (m, 1H), 4.55 (t, 1H, J = 5.9 Hz), 4, 45 - 4.41 (m, 1H), 3.85 -3.54 (m, 6H), 3.16-2.92 (m, 4H), 2.79 (t, 1H, J = 15.7 Hz), 2.68 - 2.56 (m, 3H), 1.77 (tdd, J = 6.7, 19.3, 19.3 Hz, 2H), 1.23-1.15 (m, 6H); LCMS: 538.3 [M + H] +.

Example 371 3 - ((1S, 3S) -1 - (2,6-difluoro-4 - ((1 - (3-fluoropropyl) azetidin-3-yl) oxy) phenyl) -6-fluoro-3 methyl- 1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 371.

Following the procedures of Example 368, a pair of diastereoisomers 2 were purified by chiral HPLC (ChiralPak IC, 35% IPA in heptane, 0.1% diethylamine). The second highlighted peak (rt is 14 min) is 371, isolated as a white solid (180 mg, 5%).

Other illustrative compounds of formula I in table. 2a have the following structures, corresponding names (ChemBioDraw, version 12.0.2, CambridgeSoft Corp., Cambridge, MA, USA) and biological activity. If more than one name is associated with a compound of Formula I or an intermediate, the chemical structure defines the compound.

Table 2a

No. Structure Name ER-alpha MCF7 HCS EC ₅₀ (μM) LCMS [M + H] ⁺ 431 F he RJ n7 YYV-h; hn-A ^N '^^ ^F (R) -3 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.0000943 537.3 432 _ F ^he YTVx / ^Ν τΤ (S) -3 - ((1S, 3S) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.00327 537.3 433 F ОН XTh ^f hn-X) ⁿ '^^ ^f (S) -3 - ((1R, 3R) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.0000209 537.3 434 _ F OH T GUCH J ^h fZ AnX ^{n /} ^ ^f (R) -3 - ((1R, 3S) -1- (2,6 difluoro-4 - ((1- (3- fluoropropyl) azetidine-3yl) amino) phenyl) -6fluoro-3-methyl-1,3,4,9tetrahydro-2Hpyrido [3,4-b] indol-2yl) -2-fluoro-2methylpropan-1-ol 0.000706 537.3

Example 431 (R) -3 - ((1R, 3R) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6fluoro-3 -methyl-1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 431.

Step 1: 3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-N- (1- (5-fluoro-1H-indol-3-yl) propan-2-yl) -2methylpropan-1-amine

To a solution of 1- (5-fluoro-1H-indol-3-yl) propan-2-amine (5.30 g, 26.2 mmol, 95%, prepared according to Yeung, et al., J. Med. Chem. 2010, 53, 5155-5164), in 1,4-dioxane (105 ml) cooled in an ice bath, Ν, Ν-diisopropylethylamine (6.85 ml) was added, and then [3- [tert- butyl (diphenyl) silyl] oxy-2fluoro-2-methylpropyl] trifluoromethanesulfonate (13.80 g, 28.8 mmol) in dioxane (10 ml), according to example 154, step 5. The mixture was heated at 90 ° C (bath) in for 18 h. The mixture was concentrated. Diluted Na ₂ CO ₃ was added. The contents were extracted with DCM (2x). The combined extracts were dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by flash chromatography (0-50% iPrOAc / heptane with 1% TEA (triethanolamine)) to afford the title product (10.38 g, 76%).

Steps 2-5: K- (4- (2- (3 - ((tert-butyldiphenylsilyl) oxy) -2-fluoro-2-methylpropyl) -6-fluoro-3-methyl2,3,4,9-tetrahydro- III-pyrido [3,4-b] indol-1-yl) -3,5-difluorophenyl) -1- (3-fluoropropyl) azetidin-3-amine

The specified compound was obtained in the same manner as the compound of example 145.

Step 6: racemic 3- (1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro-3methyl-1,3,4, 9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol

To a solution of n- [4 - [(1R, 3R) -2- [3- [tert-butyl (diphenyl) silyl] oxy-2-fluoro-2-methylpropyl] -6-fluoro-3methyl-1,3,4 , 9-tetrahydropyrido [3,4-b] indol-1-yl] -3,5-difluorophenyl] -1- (3-fluoropropyl) azetidin-3-amine (2.231 g, 2.879 mmol) in THF (14.4 ml) was added TBAF in THF (1.0M, 4.6 ml). The mixture was heated at 50 ° C for 24 hours. The mixture was concentrated. Diluted with iPrOAc, washed with diluted Na ₂ CO ₃ (2x) and brine, dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by flash chromatography (0-60% B / A, A: DCM, B: 20% 2M NH3 in MeOH / DCM). The collected product was subjected to chiral resolution. Stereochemistry of compounds 431-434 in the table. 2 is unknown and arbitrary.

Stage 1. Isolation of enantiomers 1 and 4.

Enantiomers 2 and 3 remain mixed. ChiralPak AD (250x30.0, 5 μm), 32.5% isocratically 0.1% NH ₄ OH in isopropanol at 150 g / min, UV-254 nm, BPR 100 bar, 40 ° C, cycle time 5 min , total time 200 min.

Stage 2. Separation of enantiomers 2 and 3.

ChiralPak OX (150x30.0, 5 μm), 30% isocratically 0.1% NH ₄ OH in methanol at 150 g / min, UV-250 nm, BPR 100 bar, temp 40 ° C, cycle time 3 min, total time 48 min.

Compounds 431-434 were characterized as follows.

Enantiomer 1: 324.8 mg. 1H NMR (400 MHz, DMSO-d ₆ ) δ 10.59 (s, 1H), 7.19-7.08 (m, 2H), 6.85-6.75 (m, 1H), 6.68 (d, J = 6.9 Hz, 1H), 6.17-6.06 (m, 2H), 5.01 (s, 1H), 4.81 (t, J = 5.8 Hz, 1H) , 4.51 (t, J = 6.1 Hz, 1H), 4.39 (t, J = 6.0 Hz, 1H), 4.33 (d, J = 4.2 Hz, OH), 3 , 99 - 3.87 (m, 1H), 3.82 - 3.72 (m, OH), 3.67 - 3.56 (m, 2H), 3.55 - 3.40 (m, 2H) , 3.19-3.05 (m, 1H), 2.95-268 (m, 4H), 1.74-1.56 (m, 2H), 1.14-0.99 (m, 6H). LCMS: 537.3 [M + H] +.

Enantiomer 2: 251.7 mg. 1H NMR (400 MHz, DMSO-de) δ 10.59 (s, 1H), 7.20-7.07 (m, 2H), 6.86-6.75 (m, 1H), 6.68 ( d, J = 6.8 Hz, 1H), 6.11 (d, J = 12.1 Hz, 2H), 5.01 (s, 1H), 4.81 (t, J = 5.8 Hz, 1H), 4.51 (t, J = 6.1 Hz, 1H), 4.39 (t, J = 6.0 Hz, 1H), 4.00 - 3.87 (m, 1H), 3, 68 - 3.57 (m, 2H), 3.55 - 3.41 (m, 2H), 3.20 - 3.06 (m, 1H), 2.95 - 2.69 (m, 4H), 1.73-1.56 (m, 2H), 1.17-0.96 (m, 6H). LCMS: 537.3 [M + H] ⁺ .

Enantiomer 3: 105.5 mg. 1H NMR (400 MHz, DMSO-de) δ 10.55 (s, 1H), 7.17-7.08 (m, 2H), 6.84-6.75 (m, 1H), 6.67 ( d, J = 6.8 Hz, 1H), 6.14-6.05 (m, 2H), 4.98 (s, 1H), 4.84 (t, J = 5.7 Hz, 1H), 4.51 (t, J = 6.0 Hz, 1H), 4.39 (t, J = 6.0 Hz, 1H), 3.99 - 3.87 (m, 1H), 3.67 - 3 , 57 (m, 2H), 3.57 -3.47 (m, 1H), 2.92-279 (m, 2H), 2.77 2.69 (m, 2H), 1.73 - 1.56 (m, 2H), 1.13-0.96 (m, 6H). LCMS: 537.3 [M + H] ⁺ .

Enantiomer 4: 151.1 mg. 1H NMR (400 MHz, DMSO-de) δ 10.55 (s, 1H), 7.18-7.07 (m, 2H), 6.84-6.75 (m, 1H), 6.67 ( d, J = 7.0 Hz, 1H), 6.10 (d, J = 12.1 Hz, 2H), 4.97 (s, 1H), 4.84 (t, J = 5.7 Hz, 1H), 4.51 (t, J = 6.1 Hz, 1H), 4.39 (t, J = 6.1 Hz, 1H), 3.98-3.89 (m, 1H), 3, 66-3.58 (m, 2H), 3.57-3.48 (m, 1H), 2.93-2.79 (m, 2H), 2.792.69 (m, 2H), 1.74 - 1.57 (m, 2H), 1.12-0.96 (m, 6H). LCMS: 537.3 [M + H] ⁺ .

Example 432 (S) -3 - ((1S, 3S) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro - 119 038835

3-methyl-1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 432.

Following the procedures of Example 431, enantiomer 432 was isolated.

Example 433. (S) -3 - ((1R, 3R) -1 - (2,6-difluoro-4 - ((1 - (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro3 -methyl-1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 433.

Following the procedures of Example 431, enantiomer 433 was isolated.

Example 434. (R) -3 - ((1R, 3S) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidin-3-yl) amino) phenyl) -6-fluoro3 -methyl-1,3,4,9-tetrahydro-2H-pyrido [3,4-b] indol-2-yl) -2-fluoro-2-methylpropan-1-ol 434.

Following the procedures of Example 431, enantiomer 434 was isolated.

Example 901. High-precision fluorescence imaging-based degradation assay of ERa in breast cancer cells.

MCF7 breast cancer cells were seeded on the first day at a density of 10,000 cells / well in a 384-well tissue culture plate coated with polylysine (Greiner # T-3101-4) in 50 μl / well RPMI medium (phenol red free) with 10 % fetal bovine serum (FBS, Fetal Bovine Serum) (purified on activated carbon) containing L-glutamine. On the second day, compounds were prepared at 2 initial compound concentrations: 100 μM and 1 μM (to ultimately obtain 2 overlapping titration curves) in a Labcyte low dead volume plate, 10 μl / well, 10 μl DMSO in designated refill wells and 5 μM fulvestant (control compound) in designated wells. Compounds and controls were dispensed using a Labcyte Echo acoustic dispenser to dispense compounds with a predetermined serial dilution (1.8x10 points in duplicate) and appropriate backfill and control compounds (final total tolerated volume was 417.5 nL and compound dispensed volume ranged from 2.5 to 417.5 nL, the final DMSO concentration was 0.84% (v / v), ultimately giving a concentration range of 0.05 to 835 nM. Cell plates were incubated at 37 ° C in for 4 hours. Fixation and permeabilization were performed using a plate washer and a Biotek EL406 dispenser as follows: The cells were fixed by adding 15 μl of 16% paraformaldehyde (Electron Microscopy Sciences No. 15710-S) directly to 50 μl of cell culture medium in each well using a peristaltic cassette pump 5 μl on Biotek EL406 (final formaldehyde concentration was 3.7% w / v) Samples were incubated for 30 min. the well holder was aspirated and 50 μl / well of phosphate buffered saline (PBS) containing 0.5% (w / v) bovine serum albumin, 0.5 (v / v) Triton X-100 was added to each well (antibody dilution buffer). Samples were incubated for 30 minutes. The contents of the wells were aspirated and washed 3 times with 100 μl / well PBS. Immunofluorescent staining for estrogen receptor α (ESR1) was performed using a plate washer and a Biotek EL406 dispenser as follows.

The supernatant was aspirated from the wells and added at 25 μl / well anti-ESRI-MKA (F10) (Santa Cruz sc-8002) diluted 1: 1000 in antibody dilution buffer. Samples were incubated for 2 h at room temperature. Samples were washed 4 times with 100 μl / well PBS. To each well was added 25 μl / well of a solution of secondary antibodies (Alexafluor 488 conjugate with anti-mouse IgG (LifeTechnologies # A21202), diluted 1: 1000, and Hoechst 33342, 1 μg / ml, diluted in antibody dilution buffer). Samples were incubated for 2 h at room temperature. Samples were washed 3 times with 100 μl / well PBS using Biotek EL406. Quantitative fluorescence imaging of ESR1 was performed using Cellomics Arrayscan V (Thermo). Fluorescence images of samples (channel 1: XF53 Hoechst (DNA staining), channel 2: XF53 FITC (ESR1 staining)) were acquired using Cellomics VTI Arrayscan and Autoexposure Bioapplication Compartmental Analysis (based on DMSO control wells), setting the maximum target percentile by 25% saturation target for both channels. Channel 1 (DNA staining) was used to determine the nuclear region (Circ). Measurements of Mean_CircAvglntCh2, which represent the fluorescence intensity of Alexafluor 488 (ESR1) in the nuclear region, were performed for each cell and averaged over all measured cells. Data analysis was performed using Genedata Screener Software using samples treated with DMSO and 5 nM fulvestrant to determine 0% and 100% changes in ESR1. The Robust Fit method was used to determine the inflection point of the curve (EC ₅₀ ) and the plateau of maximum effect (S _inf ). Degradation data for exemplary compounds of Formula I are reported as ER-α MCF7 HCS Sj _nf (%) in table. one.

Example 902. In vitro cell proliferation assay.

The efficacy of estrogen receptor modulator compounds and chemotherapeutic compounds was measured in a cell proliferation assay using the following protocol ((Mendoza et al. (2002) Cancer Res. 62: 5485-5488).

The CellTiter-Glo® Luminescent Cell Viability Assay is a homogeneous method for determining the number of viable cells in culture based on the amount of ATP present, which signals the presence of metabolically active cells. The CellTiter-Glo® assay is designed for use in a multi-well plate format, making it ideal for automated high-throughput screening (HTS), cell proliferation and cytotoxicity assays. The homogeneous assay procedure involves the addition of one reagent (CellTiter-Glo® reagent) directly to cells cultured in serum supplemented medium. The steps of cell washing, media removal, and re-pipetting are not required. The CellTiter-Glo® luminescent cell viability assay, including reagents and protocols, is commercially available (Promega Corp., Madison, WI, Technical Bulletin TB288).

The assay evaluates the ability of compounds to enter cells and inhibit cell proliferation. The principle of the assay is based on determining the number of viable cells present by quantifying the ATP present in a homogeneous assay, where the addition of Cell Titer-Glo® reagent lyses the cells and produces a luminescent signal via a luciferase reaction. The luminescent signal is proportional to the amount of ATP present.

Procedure.

Day 1 - Cell seeding plates (384 well, black, clear bottom, TC plates with cap from Falcon # 353962), harvested cells, cells are seeded at 1000 cells at 54 μL per well in 384 well cell plates for 3-day analysis. Cell culture medium: RPMI or high glucose DMEM, 10% fetal bovine serum, 2 mM L-glutamine, P / S (penicillin / streptomycin). Incubate overnight at 37 ° C, 5% CO ₂ .

Day 2 - Add drugs to cells, dilute compounds, DMSO plates (1: 2 serial dilutions to obtain 9 points). Add 20 μl of compounds at a concentration of 10 mM to the 2nd column of a 96-well plate. Perform a 1: 2 dilution series in the plate (10 μl plus 20 μl 100% DMSO) to obtain a total of 9 points using Precision Media Plates 96-well polypropylene conical bottom plates from Nunc (Cat. # 249946) (dilution 1: 50). Add 147 μl of medium to all wells. Transfer 3 μl of DMSO plus compound from each well of the DMSO plate to each corresponding well of the Media Plate using Rapidplate® (Caliper, a Perkin-Elmer Co.). To test the combination of two compounds, transfer 1.5 μl of one drug plus DMSO from each well of the DMSO plate to each corresponding well of the Media Plate using Rapidplate®. Then 1.5 μl of another drug is transferred to the plate with the medium.

Drugs are added to the cells, the plate with cells (dilution 1:10) is added 6 μl of medium plus compound directly to the cells (already 54 μl of medium with cells). Incubate for 3 days at 37 ° C, 5% CO2 in an incubator that will not be opened frequently.

Day 5 - Treat plates, thaw Cell Titer Glo Buffer at room temperature. Remove the cell plates from 37 ° C and bring to room temperature over about 30 minutes. Add Cell Titer Glo® Buffer to Cell Titer Glo® Substrate (bottle to bottle). Add 30 μl Cell Titer Glo® Reagent (Promega Cat # G7572) to each cell well. Place on a shaker for about 30 minutes. Read the luminescence on the Analyst HT Plate Reader (half a second per well).

Cell Viability Assays and Combination Assays: Cells were plated at a density of 1,000 to 2,000 cells / well in 384-well plates for 16 hours. On day two, nine serial 1: 2 dilutions of compound 1: 2 in DMSO were performed on a 96-well plate. Compounds were also diluted in growth media using a Rapidplate® robot (Zymark Corp., Hopkinton, MA, USA). The diluted compounds were then added to four replicate wells in 384-well cell plates and incubated at 37 ° C and 5% CO ₂ . After 4 days, the relative number of viable cells was measured by luminescence using Cell-TiterGlo® (Promega) according to the manufacturer's instructions and read on a Wallac MultiLabel Reader® (PerkinElmer, Foster City). _{EC 50} values were calculated using Prism® 4.0 software (GraphPad, San Diego, USA). Drugs in the combination assays were administered starting at 4xEC ₅₀ concentrations. In cases where the EC _{50 of the} drug was more than 2.5 μM, the highest concentration used was 10 μM. Estrogen receptor modulating compounds were added at the same time or 4 hours apart (one in front of the other) in all assays.

An additional illustrative in vitro cell proliferation assay includes the following steps:

1) a 100 ml aliquot of cell culture containing approximately 10 ⁴ cells (see Table 3 for cell lines and tumor type) was maintained in medium in each well of a 384-well plate with opaque walls;

2) control wells were prepared containing medium without cells;

3) compound was added to the experimental wells and incubated for 3-5 days;

4) The plates are brought to room temperature over about 30 minutes;

5) CellTiter-Glo® reagent was added to each well in a volume equal to the volume of culture medium present in each well;

6) the contents were mixed for 2 min on an orbital shaker to induce cell lysis;

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7) the plate was incubated at room temperature for 10 min to stabilize the luminescence signal;

8) luminescence was recorded and presented on the graphs in the form of RLU - relative luminescence unit;

9) were analyzed using the combined Zhou and Talaley method and dose-response analysis using CalcuSyn® software (Biosoft, Cambridge, UK) to obtain the combination index.

Alternatively, cells were plated at optimum density in a 96-well plate and incubated for 4 days in the presence of the test compound. Then, Alamar Blue ™ was added to the assay medium and the cells were incubated for 6 hours before recording with excitation at a wavelength of 544 nm, emission of 590 nm. _{EC 50} values were calculated by fitting a sigmoidal dose-response curve.

Alternatively, proliferation / viability was analyzed 48 hours after drug treatment using the CellTiter-Glo® reagent (Promega Inc., Madison, WI, USA). DMSO treatment was used as a control in all viability assays. _{IC 50} values were calculated using XL fit software (IDBS, Alameda, CA, USA).

Cell lines were obtained from either ATCC (American Type Culture Collection, Manassas, VA, USA) or DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany). The cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, 2 mM L-glutamine, and 100 mg / ml streptomycin (Life Technology, Grand Island, NY) at 37 ° C in an atmosphere with 5 % CO ₂ .

Example 903 In vitro assay of MCF7 cell proliferation.

MCF7 cells were washed with PBS and plated in RPMI 1640 (Gibco 11835-030 [β-phenol plus glutamine]) and 10% activated charcoal purified FBS (Gibco 12676-029) in polylysine-coated 384 well tissue culture plates (Greiner) at 25000 cells / ml, 40 μl / well and incubated overnight. Compounds were prepared serially in DMSO at 500-fold final desired concentration using Biomek-FX and diluted 50-fold in RPMI 1640. Control fulvestrant and negative control DMSO were also prepared in a similar manner. 5 μl of each individual compound concentration and each control compound were transferred to a cell plate. Fulvestrant was added to control wells at a final concentration of 100 nM. DMSO was added to negative control wells (0.2% v / v). To each well of the cell plate was added 5 μl of 1 nM estradiol (in RPMI 1640 without phenol red (Gibco 11835-030)) (excluding control wells without estradiol). Cells were incubated for 72 hours, then lysed using CellTiterGlo reagent (Promega # G7572), 40 μl / well, and luminescence was measured on an Envision plate reader (Perkin Elmer). Data were analyzed using Genedata Screener software using DMSO and fulvestrant treated samples to determine 0% and 100% inhibition, and EC ₅₀ values were calculated using a curve fitting method using a robust method.

Example 904 Assay of Era Coactivator Peptide Antagonist.

Test compounds were prepared in 1 mM solution in DMSO and serially diluted at 12 points by 1-3-fold titration using Biomek FX in 384-well transparent polypropylene V-bottom plates (Greiner cat. # 781280). An intermediate 3-fold dilution of the compound was prepared by mixing 1 ml of each concentration of the serial dilution of the compound with 32.3 ml of TR-FRET Coregulator Buffer E (Life Technologies PV4540). 2 ml of the 3-fold intermediate dilution of the compound was transferred to a 1536-well plate (Aurora Biotechnologies MaKO 1536 Black Plate, # 00028905) using Biomek FX. A Bioraptr Dispenser® (Beckman Coulter) was used to dispense: 2 ml / well of 3X ERa solution: 22 nM ERa (human estrogen receptor α, GST-labeled ligand binding domain ESR1 spanning residues S282-V595, or wild-type sequence , or containing mutations Y537S or D538G) in the buffer TR-FRET Coregulator Buffer E containing 7.5 mm dithiothreitol (DTT); and 2 ml of a 3-fold assay mixture (750 nM fluorescein - PGC1a peptide sequence, Life Technologies PV4421), 12 nM estradiol, 15 nM Tb-labeled anti-GST antibody in TR-FRET Coregulator Buffer E (with 7.5 mM DTT). Control wells without receptor were buffered without GST-ERa protein. The plates were centrifuged at 1800 rpm for 20 seconds in a V-plate centrifuge, and then the plates were covered and incubated for 2 hours at room temperature. Measurements were performed with a Perkin Elmer EnVision Fluorescence Reader using TR-FRET settings (upper mirror: Perkin Elmer Lance / DELFIA Dual emission (PE # 2100-4160); excitation filter: Perkin Elmer UV (TFR) 340 nm (PE # 2100 -5010); emission filter: Chroma 495 nm / 10 nm and 520 nm / 25 nm (Chroma no. PV003 filters for LanthaScreen, diameter 25 mm for EnVision); excitation light: 100%; delay: 100 μs; window time: 200; number of consecutive windows: 1; time between flashes: 2000 μs; number of flashes:

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one hundred; number of flashes (2nd detector): 100. Percent inhibition values were calculated relative to controls without compound (only with DMSO) and controls without ERa. Curve fitting and IC ₅₀ calculations were performed using Genedata Screener software.

Example 905 In vivo efficacy of murine tumor xenograft.

Mice: Female mice with severe combined immunodeficiency (Fox Chase SCID®, C.B17 / IcrHsd, Harlan) or nude mice (Taconic Farms, Harlan), 8 to 9 weeks old and weighing 15.1 to 21.4 d on day 0 of the study.

Animals received ad libitum water (reverse osmosis, 1 ppm CI) and NIH 31 Modified and Irradiated Lab Diet®, consisting of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. Mice were housed in irradiated laboratory animal cages ALPHA-Dri® bed-o'cobs® Laboratory Animal Bedding in static microisolators with a 12-hour light cycle, a temperature of 2122 ° C (70-72 ° F) and a humidity of 40-60%. The PRC, in particular, follows the guidelines of the Laboratory Animal Care and Use Guidelines regarding containment, agriculture, surgical procedures, feed and fluid regulation, and veterinary care. The PRC Animal Care and Use Program is accredited by the International Association for the Certification and Accreditation of Laboratory Animals (AAALAC), which ensures that accepted standards for the care and use of laboratory animals are adhered to.

Tumor implantation: Xenografts were initiated by cancer cells. The cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 2 mM glutamine, 100 U / ml penicillin, 100 μg / ml streptomycin sulfate, and 25 μg / ml gentamicin. Cells were harvested in exponential growth phase and resuspended in phosphate buffered saline (PBS) at a concentration of 5x106 or 10x106 cells / ml, depending on the doubling time of the cell line. Tumor cells were implanted subcutaneously in the right flank and tumor growth was monitored until the mean size approached the target range of 100 to 150 mm ³ . Twenty-one days after tumor implantation, designated study day 0, mice were divided into four groups, each of which consisted of ten mice with individual tumor volumes ranging from 75 to 172 mm ³ and average tumor volumes in the group 120-121 mm ³ (see Appendix A). The volume was calculated using the formula tumor volume (mm ³ ) w ² xl) / 2, where w is the width and l is the length of the tumor in mm.

Tumor weight can be estimated with the assumption that 1 mg is equivalent to 1 mm ³ tumor volume.

Therapeutic Agents: Estrogen receptor modulator compounds and chemotherapeutic agents are generally prepared from dry powders stored at room temperature and protected from light. Drug doses were prepared weekly in 0.5% methylcellulose: 0.2% Tween 80 in deionized water (vehicle) and stored at 4 ° C. Vehicle (+) was a diluent / buffer with ethinylestradiol (ethinylestradiol, EE2) at a dose of 0.1 mg / kg. Carrier (-) was a solvent / buffer without ethinyl estradiol. Doses of the compounds were prepared each day of dosing by diluting an aliquot of the effluent with sterile saline (0.9% NaCl). All doses were formulated to deliver the indicated dose (mg / kg) in a volume of 0.2 ml per 20 g of body weight (10 ml / kg).

Administration: All doses were scaled by body weight of individual animals and administered as indicated.

End point: tumor volume was measured in 2 dimensions (length and width) using an Ultra CaI IV meter (model 54 10 111; Fred V. Fowler Company) as follows: tumor volume (mm ³ ) = (length x width ² ) x 0 , 5, and analyzed using Excel version 11.2 (Microsoft Corporation). A linear mixed effect (LME) modeling approach has been used to analyze repeated measurement of tumor volumes in the same animals over time ((Pinheiro J., et al. Nlme: linear and nonlinear mixed effects models. R package version 3.1 92.2009; Tan N., et al. Clin. Cancer Res. 2011; 17 (6): 1394-1404) This approach addresses both repeated measurements and moderate dropouts due to any death of the animal, not treatment-related until study termination Cubic regression splines are used to approximate a non-linear profile for the change over time in log2 tumor volume at each dose level. These non-linear profiles were then related to dose within the mixed model. Tumor growth inhibition as a percentage of vehicle control ( % TGI) was calculated as the percentage of area under the curve (AUC) for the corresponding group (with the corresponding dose) per day relative to the vehicle using the following formula:% TGI = 100 x (1 - AUC _{to per} / AUC _nose .). Using this formula, a TGI value of 100% indicates tumor growth arrest, a TGI value of more than 1% but less than 100% indicates tumor growth retardation, and a TGI value of more than 100% indicates tumor regression. Partial response (PR) for an animal is defined as tumor regression of more than 50%, but less than 100% of the initial tumor volume. Complete response (CR) was defined as 100% tumor regression (i.e., unmeasurable tumor) on any day of the study.

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Toxicity: Animals were weighed daily for the first five days of the study and twice a week thereafter. Animal body weights were measured using the Adventurer Pro® AV812 scale (Ohaus Corporation). The percentage change in body weight was calculated as follows: change in body weight (%) = weight _{new day} - weight _{day 0} ) / weight _{day 0} ] x 100 clinical manifestations of toxicity were recorded. Acceptable toxicity was defined as the mean body weight loss (BW) of less than 20% in the study group and no more than one treatment-related death (TR) in ten treated animals. Any dosing regimen that resulted in greater toxicity was considered a maximum tolerated dose (MTD). Death was classified as TR if it was due to side effects of treatment as evidenced by clinical signs and / or autopsy, or could also be classified as TR if it occurred for unknown reasons during the dose period or within 10 days after the last dose. ... Death was classified as NTR if there was no evidence that the death was associated with side effects of treatment.

In vivo breast cancer xenograft model; (MCF-7, tamoxifen sensitive): Time-release beads containing 0.72 mg of 17-β-estradiol were implanted subcutaneously in nu / nu mice. MCF-7 cells were grown in RPMI medium containing 10% FBS at 5% CO ₂ , 37 ° C. Trypsinized cells were plated in beads and resuspended in 50% RPMI (serum free) and 50% Matrigel at a density of 1x10 ⁷ cells / ml. MCF-7 cells were injected subcutaneously (100 μl / animal) into the right flank 2-3 days after implantation of the beads. The tumor volume (length x width ^2/2) was monitored every two weeks. When the tumors reached an average volume of about 200 mm ³ , the animals were randomized and the administration began. The animals were dosed with vehicle or compound daily for 4 weeks. Tumor volume and body weight were monitored every two weeks throughout the study.

In vivo breast cancer xenograft model; (tamoxifen resistant model): nu / nu mice (with 17-β-estradiol beads injected; 0.72 mg, 60-day slow release) with MCF-7 tumors (mean tumor volume 200 mm ³ ) were injected with tamoxifen ( citrate) through a gastric tube. The tumor volume (length x width ^2/2) and body weight were monitored twice per week. After a significant antitumor response in which the tumor volume remained static, apparent tumor growth was initially observed after about 100 days of treatment. After 120 days of treatment, the dose of tamoxifen was increased. Rapidly growing tumors were considered resistant to tamoxifen and selected for in vivo passage to new host animals. Fragments of tamoxifen-resistant tumors (approximately 100 mm ³ / animal) were implanted subcutaneously in the right flank of female nu / nu mice (with 17-β-estradiol beads (0.72 mg; 60-day slow release)). Passaged tumor was maintained under constant selection on tamoxifen, and tumor volume (length x width ^2/2) was monitored weekly. When the tumor volume reached approximately 150-250 mm ³ , the animals were randomized into groups (mean tumor volume 200 mm ³ ) and the administration of tamoxifen was stopped. The animals were dosed with vehicle or compound daily for 4 weeks. Tumor volume and body weight were monitored twice a week throughout the study.

Example 906. Analysis of the total mass (wet) of an immature uterus.

Female immature CD-IGS rats (21 days of age upon arrival) were dosed with the compounds for three days. The animals were dosed daily for three days. In the antagonist mode, the vehicle or test compound was orally administered by gavage, followed by oral administration of ethinylestradiol at a dose of 0.1 mg / kg 15 minutes later. In agonist mode, the vehicle or test compound was administered orally by gavage. On the fourth day, 24 hours after dosing, plasma was collected for pharmacokinetic analysis. Immediately after collecting plasma, the animals were euthanized and the uterus removed and weighed.

Uterus and ovaries from two animals from each group were fixed in 10% neutral buffered formalin and embedded in paraffin, sectioned and stained with hematoxylin-eosin (SDPath). The stained tissue was analyzed by a certified pathologist. Uterus and ovaries from four animals from each group were frozen in liquid nitrogen for transcriptional analysis, examining a selected set of genes modulated by the estrogen receptor.

Mice were injected with compounds of formula I (1R, 3R) -1- (2,6-difluoro-4 - ((1- (3-fluoropropyl) azetidine-3yl) oxy) phenyl) -2- (2-fluoro-2-methylpropyl ) -3-methyl-2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 101 and (1R, 3R) -1- (2,6-difluoro-4- (2- ( 3- (fluoromethyl) azetidin-1-yl) ethoxy) phenyl) -2- (2-fluoro-2-methylpropyl) -3methyl-2,3,4,9-tetrahydro-1H-pyrido [3,4-b] indole 102, tamoxifen, fulvestrant, AZD9496 (WO 2014/191726, example 1, page 74, US 9155727) and two controls: vehicle and vehicle plus ethinyl estradiol (EE). All compounds were administered orally, QDx3. Calculated the ratio of the total weight of the uterus (Uterine Wet Weight, UWW) to body weight. The average height of the endometrium in the transverse sections of the uterus was measured by histology. The height of endometrial cells was measured from the basement membrane to the apical (luminal) surface using a slide projector with 20X magnification. Areas with an oblique cut were avoided. In the UWW assay in agonist mode, compounds 101 and 102 of formula I are antagonists, while AZ49496 is a partial agonist.

Example 907. 10-day analysis of the total weight of the uterus of adult rats.

Female CD-IGS rats (69 days old, Charles River Laboratories) were purchased and divided into groups. Group 1 underwent oophorectomy from a supplier (Charles River Laboratories) at the age of 60 days, and the study began 2 weeks after surgery, while groups 2-8 remained intact. The vehicle or test compound was administered orally for 10 days. Two hours after the 10th and last dose, heart punctures were performed and blood serum was collected for pharmacokinetic and estradiol analysis. Immediately after collecting the serum, the animals were euthanized, the uterus and ovaries were removed and weighed.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, these descriptions and examples should not be construed as limiting the scope of the invention. The disclosure of all patent and scientific literature cited herein is expressly incorporated by reference in its entirety.

Claims (22)

CLAIM 1. Compound of formula I and its stereoisomers or pharmaceutically acceptable salts, where Y ¹ represents CRb or N; Y ² represents - (CH ₂ ) -; Y ³ represents NRa or C (R ^b ) ₂ ; where one of Y ¹ or Y ³ represents N or NRa; R ^{a is} independently H, C ₁ -C ₆ alkyl, C ₂ -C ₈ alkenyl, propargyl, C ₃ -C ₆ cycloalkyl or C ₃ -C ₆ heterocyclyl optionally substituted with one or more groups independently selected from F, Cl, Br, I, CN, OH, OCH3 or SO2CH3; Rb independently represents H, C ₁ -C ₆ -alkyl, C ₂ -C ₈ -alkenyl, propargyl, - (C1-C ₆ -alkyldiyl) (C ₃ -C ₆ -cycloalkyl), C ₃ -C ₆ -cycloalkyl or C ₃ -C ₆ heterocyclyl optionally substituted with one or more groups independently selected from F, Cl, Br, I, CN, —CH2F, —CHF2, —CF3, —CH2CF3, —CH2CHF2, —CH2CH2F, OH, OCH ₃ or SO ₂ CH ₃ ; R ^c represents H; Z ¹ represents a bond; Cy is phenyldiyl or 5-6 membered heteroaryldiyl; Z ² represents O or O- (C ₁ -C ₆ -alkylduyl); R ¹ and R ² are H; R ³ and R ^{4 are} independently H, -CH ₃ , -CH ₂ CH ₃ , -CH (CH ₃ ) ₂ , -CH ₂ CH (CH ₃ ) ₂ , -CH2OH, -CH2OCH3, -CH2CH2OH, -C ( CH3) 2OH, -CH (OH) CH (CH3) 2, -C (CH3) 2CH2OH, -CH2CH2SO2CH3, -CH ₂ OP (O) (OH) ₂ , -CH2F, -CHF2, -CH2NH2, -CH ₂ NHSO ₂ CH ₃ , —CH2 NHCH3, —CH ₂ N (CH ₃ ) ₂ , —CF ₃ , —ch ₂ cf ₃ , —CH2 CHF2, —CH (CH3) CN, —C (CH3) 2CN, or —CH2 CN; R ⁵ is H or C1-C9 alkyl optionally substituted with one or more of halogen, CN, OR ^a , N (R ^a ) 2, C1-C ₉ alkyl, C ₃ -C ₉ cycloalkyl, C ₃ -C ₉ heterocycle, C ₆ -C ₉ aryl, C (O) R ^b , C (O) NR ^a , SO2R ^a or SO2NR ^a ; R ⁶ is F, Cl, Br, I, —CN, or —CH3; and m is 0 or 1; where alkyldiyl, phenyldiyl and heteroaryldiyl are optionally substituted with one or more groups independently selected from F, Cl, Br, I, —CN, —CH ₃ , —CH ₂ CH ₃ , —CH (CH ₃ ) ₂ , - CH ₂ CH (CH ₃ ) ₂ , -CH ₂ OH, -CH2OCH3, -CH2CH2OH, -C (CH3) 2OH, -CH (OH) CH (CH3) 2, -C (CH3) 2CH2OH, -CH2CH2SO2CH3, -CH2OP (O) (OH) 2, -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CHF2, -CH2CH2F, -CH (CH3) CN, -C (CH3) 2CN, -CH2CN, -CH2NH2, -CH2NHSO2CH3, - CH2NHCH3, -CH2N (CH3) 2, -CO2H, -COCH3, -CO2CH3, -CO2C (CH3) 3, -COCH (OH) CH3, -CONH2, -CONHCH3, -CON (CH3) 2, -C (CH3) 2CONH2, -NH2, -NHCH3, -N (CH3) 2, -NHCOCH3, -N (CH3) COCH3, -NHS (O) 2CH3, -N (CH ₃ ) C (CH ₃ ) 2CONH ₂ , -N (CH3 ) CH2CH2S (O) 2CH3, -NO2, = O, -OH, -OCH3, -OCH2CH3, -OCH2CH2OCH3, -OCH2CH2OH, -OCH2CH2N (CH3) 2, -OP (O) (OH) 2, -S (O) 2N (CH3) 2, -SCH3, -S (O) 2CH3, -S (O) 3H, cyclopropyl, cyclopropylamide, cyclobutyl, oxetanyl, azetidinyl, (1methylazetidin-3-yl) oxy, N-methyl-N-oxetane- 3-ulamino, azetidin-1-ylmethyl, benzyloxyphenyl, pyr-125 038835 rolidin-1-yl, pyrrolidin-1-yl-methanone, piperazine-1 -yl, morpholinomethyl, morpholinomethanone and morpholino; where heteroaryldiyl means a divalent aromatic radical containing 5- or 6-membered rings, including fused ring systems (at least one of which is aromatic) of 5-20 atoms, containing one or more than one heteroatom independently selected from nitrogen, oxygen and sulfur ; where heterocycle and heterocyclyl denote a monocyclic or bicyclic saturated or partially unsaturated carbocyclic radical containing from 3 to 9 ring atoms, in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and sulfur, and the remaining ring atoms are S. 2. A compound according to claim 1 having formula Ia 3. The compound according to claim 2, having the formula Ib where R ⁷ represents F, Cl, Br, I, -CN or -CH ₃ ; and n is 0, 1, or 2. 4. A compound according to claim 1 having the formula Ic or a pharmaceutically acceptable salt thereof, wherein Z ² is OC ₁ -C ₆ -alkyldiyl). 5. A compound according to claim 4 having the formula Id or a pharmaceutically acceptable salt thereof, wherein Z ² is O- (C ₁ -C ₆ -alkyldiyl). 6. A compound according to claim 5 having formula Ie 126,038835 or a pharmaceutically acceptable salt thereof, wherein R ⁸ is H or —CH3. 7. A compound according to claim 1, wherein Y ¹ is CR ^b and Y ³ is NR ^a . 8. A compound according to claim 1, wherein Y ¹ is N and Y ³ is C (R ^b ) 2. 9. A compound according to claim 1, wherein Cy is phenyldiyl which is substituted with one or more than one F. 10. A compound according to claim 1, wherein R ³ is H and R 4 is —CH 3. 11. A compound according to claim 1 wherein R ⁵ is C ₁ -C ₆ fluoroalkyl. 12. The compound of claim 1, wherein m is 0. 13. A compound according to claim 1 having the formula - 127 038835 - 128 038835 - 129 038835 - 130 038835 - 131 038835 - 132 038835 - 133 038835 - 134 038835 - 135 038835 - 136 038835 or a pharmaceutically acceptable salt thereof. 14. A compound according to claim 1 having the formula - 137 038835 or a pharmaceutically acceptable salt thereof. 15. A compound according to claim 1, which is a compound of the formula F or F; or a pharmaceutically acceptable salt thereof. 16. A compound according to claim 1, which is a compound of the formula - 138 038835 or a pharmaceutically acceptable salt thereof. 17. The compound of claim 1, which is a compound of the formula, or a pharmaceutically acceptable salt thereof. 18. A compound according to claim 1 having the formula - 139 038835 - 140 038835 - 141 038835 - 142 038835 - 143 038835 or a pharmaceutically acceptable salt thereof. or a pharmaceutically acceptable salt thereof. 19. A compound according to claim 1 having the formula - 144 038835 20. A pharmaceutical composition having an estrogen receptor modulator activity, consisting of a compound or a pharmaceutically acceptable salt thereof according to claim 1 and at least one pharmaceutically acceptable excipient. 21. A method of treating breast cancer, lung cancer, ovarian cancer, endometrial cancer, prostate cancer or uterine cancer in a patient having such cancer, comprising administering to the patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof at π. one. 22. The method of claim 21, wherein the cancer is breast cancer.