EA047929B1 - JOINT HETEROCYCLIC DERIVATIVES - Google Patents

Description

Field of technology to which the invention relates

This application relates to compounds of fused heterocyclic derivatives, pharmaceutical compositions containing such compounds, chemical processes for preparing such compounds and their use in the treatment of diseases associated with infection caused by HBV.

Related applications

This application claims priority from European Patent Application No. 19176933.0, filed May 28, 2019, PCT/CN2020/085720, filed April 20, 2020, and U.S. Provisional Patent Application No. 62/853533, filed May 28, 2019, the contents of which are incorporated herein in their entirety.

State of the art

Chronic hepatitis B virus (HBV) infection is a major global health problem affecting more than 5% of the world's population (more than 350 million people worldwide and 1.25 million individuals in the United States).

Despite the availability of a prophylactic HBV vaccine, the burden of chronic HBV infection remains a major unmet global health need due to suboptimal treatment options and persistent rates of new infections in much of the developing world. Current treatments are not curative and are limited to only two classes of agents (interferon alpha and nucleoside analogues/viral polymerase inhibitors); drug resistance, low efficacy, and tolerability issues limit their impact. Low HBV cure rates are explained, at least in part, by the difficulty of achieving complete suppression of viral replication with a single antiviral agent. However, sustained suppression of HBV DNA slows the progression of liver disease and helps prevent hepatocellular carcinoma. Current treatment goals for HBV-infected patients are to reduce serum HBV DNA levels to low or undetectable levels and ultimately to reduce or prevent the development of cirrhosis and hepatocellular carcinoma.

The HBV capsid protein has important functions during the viral life cycle. HBV capsid/core proteins form metastable viral particles or protein coats that protect the viral genome during cell-to-cell transit and also play a central role in viral replication processes including genome encapsidation, genome replication, morphogenesis, and virion egress. Capsid structures also respond to environmental stimuli by mediating the removal of the coat after viral entry. Accordingly, appropriate timing of capsid assembly and disassembly, adequate capsid stability, and core protein function have been found to be critical for viral infectivity.

The essential function of the HBV capsid proteins places strong evolutionary constraints on the viral capsid protein sequence, resulting in the observed low sequence variability and high conservation. Accordingly, mutations in the HBV capsid that interfere with its assembly are lethal, and mutations that compromise capsid stability greatly impair viral replication. The high functional constraints on the multifunctional HBV core/capsid protein are consistent with the high sequence conservation, as many mutations are detrimental to function. Indeed, the core/capsid protein sequences are more than 90% identical among HBV genotypes and exhibit only a small number of polymorphic residues. Selection for resistance to HBV core/capsid protein binding compounds can therefore be challenging without greatly affecting the ability of the virus to replicate.

Reports describing compounds that bind viral capsids and inhibit replication of HIV, rhinovirus, and HBV provide compelling pharmacological proof of concept for the use of viral capsid proteins as targets for antiviral drugs.

There is a need in the art for therapeutic agents that can enhance the suppression of viral replication and that can treat, reduce the intensity and/or prevent infection caused by HBV. The administration of such therapeutic agents to a patient infected with HBV, either as a monotherapy or in combination with other HBV treatments or adjuvant treatments, will result in a significant reduction in viral load, improved prognosis, reduced disease progression and improved seroconversion rates.

Given the clinical importance of HBV, the identification of compounds that can enhance the suppression of viral replication and that can treat, ameliorate and/or prevent HBV infection represents an attractive approach to the development of new therapeutic agents. Such compounds are provided in this document.

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Brief description

The invention is directed to general and preferred embodiments defined respectively by the independent and dependent claims appended hereto, which are incorporated herein by reference. The present invention is directed to compounds capable of modulating capsid assembly. The compounds of the present invention may provide an appropriate balance of properties relative to the compounds of the prior art, for example, they may exhibit a different profile, exhibit improved solubility, etc. Thus, in particular, the present invention is directed to a compound of formula (I) % ^w x

N-N I

Jy-z

O'^R ¹ (I) or a stereoisomer thereof, wherein R ¹ is a 5-10 membered monocyclic or bicyclic ring, wherein the 5-10 membered monocyclic or bicyclic ring optionally contains 1-3 heteroatoms, wherein each of the heteroatoms is independently selected from N, O and S; and/or is optionally substituted with one or more substituents, each of which is independently selected from halogens, CN, CF ₃ , CF2H, CFH2, CF2CH3, C _1-6 alkyl, OC _1-6 alkyl, OCF ₃ , OCF2H and C _3-4 cycloalkyl;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, OH, C _1-4 alkyl optionally substituted with OH or F, COX _1-4 alkyl, COX _3-6 cycloalkyl, K ^ O ₂ C _1-4 alkyl and K ^ _{O 2} C _3-6 cycloalkyl;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H,

C _1-4 alkyl,

C _1-4 alkyl substituted with one or more F,

C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two, three or four heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents each independently selected from the group consisting of halogens, CN, CF ₃ , CF2H, CFH2, CF ₂ CH ₃ , CH2CF3, C _1-4 alkyl, OH, OC _1-4 alkyl, OCF3, OCF2H, SO2N(CH3)2 and C3-4 cycloalkyl; and

Z is selected from the group consisting of C(=O) and C(=S), or a pharmaceutically acceptable salt thereof,

In embodiments, the compounds of formula (I) are compounds selected from the types of compounds described or illustrated in the detailed description below.

The present invention is also directed to pharmaceutical compositions comprising one or more compounds of formula (I), pharmaceutically acceptable salts of compounds of formula (I), pharmaceutically acceptable prodrugs based on compounds of formula (I) and pharmaceutically active metabolites of formula (I). The pharmaceutical compositions may further comprise one or more

- 2 047929 how many pharmaceutically acceptable excipients or one or more other agents or therapeutic agents.

The present invention is also directed to methods of using or uses of the compounds of formula (I). In embodiments, the compounds of formula (I) are used to treat or ameliorate hepatitis B virus (HBV) infection, enhance suppression of HBV production, interfere with HBV capsid assembly or other steps of HBV viral replication or products. The methods comprise administering to a subject in need of such a method an effective amount of at least one compound of formula (I), pharmaceutically acceptable salts of the compounds of formula (I), pharmaceutically acceptable prodrugs of the compounds of formula (I), and pharmaceutically active metabolites of the compounds of formula (I). Additional embodiments of the methods of treatment are provided in the detailed description.

The object of the present invention is to overcome or reduce at least one of the disadvantages of the conventional techniques and/or the prior art or to provide an advantageous alternative thereto. Additional embodiments, distinctive features and advantages of the present invention will be apparent from the further detailed description and from the practical application of the disclosed subject matter.

Detailed description

Additional embodiments, features and advantages of the subject matter of the present invention will be apparent from the following detailed description of the present invention and its practical application. For the sake of brevity, publications, including patents, cited in this specification are incorporated herein by reference.

This document provides compounds of formula (I) and their pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs thereof, and pharmaceutically active metabolites of the disclosed compounds.

In one embodiment, provided herein are compounds of formula (I)

OR ¹ (O or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ¹ is phenyl substituted with one or more substituents, in particular one to three, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, C _1-4 alkyl, OC _1-6 alkyl, OCF3, OCF2H and C _3-4 cycloalkyl; or a 9-membered bicyclic ring containing 1 or 2 N atoms and optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, Cf ₃ , CF2H, CFH2, CF2CH3, C _1-6 alkyl, OC _1-6 alkyl, OCF3, OCF2H and C _3-4 cycloalkyl;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, C _1-4 alkyl optionally substituted with OH or F, CONHC _1-4 alkyl, CONHC _3-6 cycloalkyl, NHSO ₂ C _1-4 alkyl and NHSO ₂ C _3-6 cycloalkyl;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H,

C _1-4 alkyl,

C _1-4 alkyl substituted with one or more F,

C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two, three or four heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, CH2CF3, C^alkyl, OH, OC^alkyl, OCF3, OCF2H, SO2NCI[,)2 and C _3-4 cycloalkyl; and

Z is selected from the group consisting of C(=O) and C(=S).

In a further embodiment, the present invention provides compounds

- 3 047929 of formula (I), and their stereoisomers, and their pharmaceutically acceptable salts, described in this document, where

R ¹ is a 5-10 membered monocyclic or bicyclic ring, more particularly a 5-9 membered monocyclic or bicyclic ring, wherein the 5-10 membered monocyclic or bicyclic ring, more particularly a 5-9 membered monocyclic or bicyclic ring optionally contains 1-3 heteroatoms, wherein the heteroatoms are independently selected from N, O and S; and/or is optionally substituted with one or more substituents selected from halogens, CN, CF ₃ , CF2H, CFH2, CF2CH3, ^-alkyl, OC _1-4 alkyl, OCF ₃ , OCF2H and ^-dicloalkyl;

more particularly R ¹ is phenyl substituted with one or more substituents selected from the group consisting of Cl, F, CN, CH ₃ , CF ₃ and CF2H;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, OH, C _1-4 alkyl optionally substituted with OH or F, CONHC _1-4 alkyl, CONHC _3-6 cycloalkyl, NHSO ₂ C _1-4 alkyl and NHSO ₂ C _3-6 cycloalkyl;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H,

C _-4 alkyl,

C^alkyl substituted with one or more F,

C^alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, _CF3 , CF2H, CFH2, CF2CH3, C _- 4alkyl, _OC1-4alkyl , _OCF3 , OCF2H and N-alkyl; and

Z is selected from the group consisting of C(=O) and C(=S).

In a further embodiment, the present invention provides a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ¹ is phenyl substituted with one or more substituents selected from the group consisting of Cl, F, CN, CH3, CF3 and CF2H;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, C _1-4 alkyl optionally substituted with OH or F, CONHC _1-4 alkyl and CONHC _3-6 cycloalkyl, NHSO ₂ C _1-4 alkyl _; NHSO ₂ C _3-6 cycloalkyl;

X is selected from the group consisting of _CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H,

C _-4 alkyl,

C^alkyl substituted with one or more F,

C^alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, _CF3 , CF2H, CFH2, CF2CH3, C _- 4alkyl, _OC1-4alkyl , _OCF3 , OCF2H and N-alkyl; and

Z is selected from the group consisting of C(=O) and C(=S).

In a further embodiment, the present invention provides a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ¹ is phenyl substituted by one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogen, in particular Cl or F, CN, alkyl, in particular CH ₃ , CH2F, CHF2 and CF3H;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is equal to 0;

W represents CHR ³ ;

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R ³ is selected from the group consisting of H, F, C _1-4 alkyl optionally substituted with OH or F, and NHSO2C'i _-4 a.iKn.ia;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

C^alkyl, in particular C^alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, indolyl, imidazo[1,2-a]pyridinyl and quinolinyl;

wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, _CF3 , CF2H, CFH2, CF2CH3, _C1-4alkyl , _OC1-4alkyl , _OCF3 , OCF2H and _{C3-4cycloalkyl} ; and

Z is selected from the group consisting of C(=O) and C(=S).

In a further embodiment, the present invention provides a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein

R ¹ is phenyl substituted by one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogen, in particular Cl or F, CN, C _1-6 alkyl, in particular CH ₃ , CH2F, CHF2, CF ₃ and CF2CH3;

R ² is selected from the group consisting of H, C _1-4 alkyl, and C _1-4 alkyl substituted with one or more F;

n is equal to 0;

W represents CHR ³ ;

R ³ is selected from the group consisting of H, F, C _1-4 alkyl optionally substituted with OH or F, and M^O ₂ C _1-4 alkyl;

X is selected from the group consisting of _CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

C _1-4 alkyl, in particular C _1-2 alkyl, substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, indolyl, imidazo[1,2-a]pyridinyl and quinolinyl;

wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, _CF3 , _CF2H , _CFH2 , _CF2CH3 , _C1-4 alkyl, _{OC1-4 alkyl} , _OCF3 , OCF2H and _C3-4 cycloalkyl; and

Z is selected from the group consisting of C(=O) and C(=S).

In a still further embodiment, the present invention provides a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ¹ is phenyl substituted by one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogen, in particular Cl or F, CN, alkyl, in particular CH ₃ , CH2F, CHF2, CF ₃ and CF2CH3;

R ² is H or methyl, in particular methyl;

n is equal to 0;

W represents CHR ³ ;

R ³ is H;

X is CH2;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

C^alkyl, in particular C^alkyl substituted with a 5-6-membered monocyclic aromatic ring optionally containing one, two or three heteroatoms, each independently selected from N, O and S, and selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl and oxadiazolyl;

wherein said 5-6 membered monocyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, _CF3 , CF2H, CFH2, CF2CH3, ^^alkyl, OCl _- 4alkyl,

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OCF3, OCF2H and C _3-4 cycloalkyl; and

Z is C(=O).

In embodiments, the compound of formula (I) is a compound wherein R ¹ is phenyl substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of substituents representing fluoro, chloro and cyano, more particularly wherein R ¹ is dichlorophenyl, 3-cyano-4-chlorophenyl or 3-cyano-4-fluorophenyl.

In embodiments, the compound of formula (I) is a compound wherein R ¹ is phenyl substituted with one or more substituents that are chloro, more particularly wherein R ¹ is dichlorophenyl.

In embodiments, the compound of formula (I) is a compound wherein R ² is H or methyl.

In embodiments, the compound of formula (I) is a compound wherein Z is C=O.

In embodiments, the compound of formula (I) is a compound wherein X is CH ₂ .

In embodiments, the compound of formula (I) is a compound wherein W is _CH2 or CHF.

In embodiments, the compound of formula (I) is a compound wherein R ⁵ is selected from the group consisting of _C1-4 alkyl and _C1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, _CF3 , CF2H, CFH2, CF2CH3, _C1-4 alkyl, _OC1-4 alkyl, _OCF3 , OCF2H and _C3-4 cycloalkyl.

In embodiments, the compound of formula (I) is a compound wherein R ⁵ is C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are N, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, C _1-4 alkyl, OC _1-4 alkyl, OCF ₃ , OCF2H and C _3-4 cycloalkyl.

In embodiments, the compound of formula (I) is a compound wherein R ⁵ is C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring is selected from the group consisting of phenyl, pyridyl, pyrazolyl or indolyl, and wherein the 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, CF ₃ , CF2H, CFH2, CF2CH3, C _1-4 alkyl, OC _1-4 alkyl, OCF ₃ , OCF2H and C _3-4 cycloalkyl.

In embodiments, the compound of formula (I) is a compound wherein R ⁵ is C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring is selected from the group consisting of phenyl, pyridyl, pyrazolyl or indolyl, and wherein the 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, _C1-4 alkyl, OCl _-4 alkyl, OCF3 and OCF2H.

In embodiments, the compound of formula (I) is a compound as described herein, wherein Y is NR ⁵ ; wherein

R ⁵ is selected from the group consisting of

_C1-4 alkyl, in particular _C1-2 alkyl, substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two, three or four heteroatoms, each of the heteroatoms being independently selected from N, O and S, and selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, tetrazolyl, indolyl, imidazo[1,2,4]pyridinyl, quinolinyl and [1,2,4]triazolo[1,5n]pyrimidinyl;

wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, _C1-4 alkyl, OCl _-4 alkyl, _OCF3 , OCF2H and _C3-4 cycloalkyl.

In embodiments, the compound of formula (I) is a compound as described herein, wherein Y is NR ⁵ ; wherein

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R ⁵ is selected from the group consisting of

C1-4alkyl, in particular C1-4alkyl, substituted with a 5-6 membered monocyclic aromatic ring optionally containing one, two, three or four heteroatoms each independently selected from N, O and S and selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl and tetrazolyl;

wherein said 5-6 membered monocyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, C alkyl, OCF3, OCF2H and C _3-4 cycloalkyl.

In embodiments, the compound of formula (I) is a compound as described herein, wherein Y is NR ⁵ ; wherein

R ⁵ is selected from the group consisting of ^^alkyl, in particular ^^alkyl substituted with a 5-6-membered monocyclic aromatic ring optionally containing one, two or three heteroatoms, each independently selected from N, O and S, and selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, triazolyl and oxadiazolyl;

wherein said 5-6 membered monocyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, N-alkyl, OCF3, OCF2H and N-dicloalkyl.

In embodiments, the compound of formula (I) is a compound as described herein, wherein Y is NR ⁵ ; wherein

R ⁵ is selected from the group consisting of ^^alkyl, in particular ^^alkyl substituted with a 5-6-membered monocyclic aromatic ring optionally containing one, two or three N atoms;

wherein said 5-6 membered monocyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, alkyl, OC _1-4 alkyl, OCF3, OCF2H and cycloalkyl.

In embodiments, the compound of formula (I) is a compound as described herein, wherein Y is NR ⁵ ; wherein

R ⁵ is selected from the group consisting of ^^alkyl, in particular ^^alkyl substituted with a 5-6-membered monocyclic aromatic ring, optionally containing one, two or three N atoms, selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl and triazolyl;

wherein said 5-6 membered monocyclic aromatic ring is optionally substituted with one or more substituents, in particular one to three substituents, each independently selected from the group consisting of halogens, CN, CF3, CF2H, CFH2, CF2CH3, alkyl, OC _1-4 alkyl, OCF3, OCF2H and cycloalkyl.

In embodiments, the compound of formula (I) is a compound that exhibits an EC ₅₀ value of less than 0.10 μM for inhibiting HBV DNA in the hepG2.117 cell line.

A further embodiment of the present invention is a compound selected from the group consisting of the compounds described below (see Table 1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

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Table 1

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Pharmaceutical compositions.

Also disclosed in this document are pharmaceutical compositions comprising (A) at least one compound of formula (I)

R ²

(I)

- 10 047929 or a stereoisomer thereof, wherein R ¹ is a 5-10 membered monocyclic or bicyclic ring, more particularly a 5-9 membered monocyclic or bicyclic ring, wherein the 5-10 membered monocyclic or bicyclic ring, more particularly a 5-9 membered monocyclic or bicyclic ring optionally contains 1-3 heteroatoms, each of the heteroatoms being independently selected from N, O and S; and/or optionally substituted with one or more substituents each independently selected from halogens, CN, CF ₃ , CF2H, CFH2, CF2CH3, C ₁ . ₆ alkyl, OC ₁ . ₆ alkyl, OCF ₃ , OCF2H and C ₃ . ₄ cycloalkyl;

more particularly R ¹ is phenyl substituted with one or more substituents each independently selected from the group consisting of Cl, F, CN, CH ₃ , CF ₃ and CF2H;

R ² is selected from the group consisting of H, C ₁ . ₄ alkyl, and C ₁ . ₄ alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, OH, C ₁ . ₄ alkyl optionally substituted with OH or F, CONHC ₁ . ₄ alkyl, CONHC ₃ . ₆ cycloalkyl, NHSO ₂ C ₁ . ₄ alkyl and NHSO ₂ C ₃ . ₆ cycloalkyl;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H,

C ₁ . ₄ alkyl,

C ₁ . ₄ alkyl substituted with one or more F,

C _1.4 alkyl substituted with a _5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two, three or four heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents each independently selected from the group consisting of halogens, CN, CF ₃ , CF2H, CFH2, CF2CH3, CH2CF3, C _{1.4 alkyl, OH, OC 1.4 alkyl ,} OCF ₃ , OCF2H, SO2N(CH ₃ )2 and N-cycloalkyl; and

Z is selected from the group consisting of C(=O) and C(=S), or a pharmaceutically acceptable salt thereof, and (B) at least one pharmaceutically acceptable excipient.

Also disclosed in this document are pharmaceutical compositions comprising:

(A) at least one compound of formula (I)

where R ¹ is phenyl substituted with one or more substituents selected from the group consisting of Cl, F, CN, CH ₃ , CF ₃ and CF ₂ H;

R ² is selected from the group consisting of H, C ₁ . ₄ alkyl, and C ₁ . ₄ alkyl substituted with one or more F;

n is an integer 0 or 1;

W represents CHR ³ or C=CH ₂ ;

R ³ is selected from the group consisting of H, F, CONHC ₁ . ₄ alkyl and CONHC ₃ . ₆ cycloalkyl;

X is selected from the group consisting of CH2 and O;

Y represents NR ⁵ ;

R ⁵ is selected from the group consisting of

H, ^^alkyl,

C ₁ . ₄ alkyl substituted by one or more F,

C ₁ . ₄ alkyl substituted with a 5- to 10-membered monocyclic or bicyclic aromatic ring, wherein the 5- to 10-membered monocyclic or bicyclic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5- to 10-membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogen

- 11 047929 new, CN, CF3, CF2H, CFH2, CF2CH3, C _1-4 alkyl, OC _1-4 alkyl, OCF3, OCF2H and C _3-4 cycloalkyl; and

Z is selected from the group consisting of C(=O) and C(=S);

or a stereoisomer or tautomer thereof or a pharmaceutically acceptable salt thereof and (B) at least one pharmaceutically acceptable excipient.

One embodiment of the present disclosure is a pharmaceutical composition comprising at least one pharmaceutically acceptable excipient and at least one compound described herein, for example selected from the group consisting of the compounds described below (see Table 3), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

Table 3

- 12 047929

- 13 047929

nn-^ ID .- ^Ν -, .....lm /ii |Ή ° ci-'Ay o Cl 31 nA T ₁ D_D./A' .....Ш / ίί CS) O ' /TVA ci-TZ ⁰ Cl 32 _c ,X\nCX^ ° н 33 ⁰ c/ ^N 34 ^C| X^ ⁰ 36 ><>- 11 O \ / ⁰⁰ A OQ o Q y so / O O o=\ <o / Ϊ \ w __J _C1 X\€с>3 ^{0 40} τι I1n JI _=/ ^N ^y-NHSO ₂ Me ' I j ⁴¹ V ^R> T ^N \^ ^a Ί Ή Me 42 .M Vksо V/ 5 A о о o o A t A> w Ξ-Ζ I to\3 U ^CI T1 ^r T ^N N-\ > ⁰ <rr 45 ^N - AT θ _Η ⁰ cry 46 ο,ΑΑ,ν .. ⁰ 0¼° Me 47 Ck l ΥΊι rr ^N ciAJCn^JEn-x 0 s ⁷ N ^S H 48

The pharmaceutical composition may also contain at least one additional active or therapeutic agent. Additional active therapeutic agents may include, for example, an anti-HBV agent such as an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a viral maturation inhibitor, a capsid assembly modulator, a reverse transcriptase inhibitor, an immunomodulatory agent such as a TLR agonist, or any other agents that affect the HBV life cycle and/or the consequences of infection caused by HBV. The active agents are used alone or in combination with one or more additional active agents to formulate the pharmaceutical composition of the present invention.

As used herein, the term composition or pharmaceutical composition refers to a mixture of at least one compound useful in the present invention with a pharma

- 14 047929 a ceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. There are many methods of administering the compound in the art, including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, inhalation, and topical administration.

As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, involved in carrying or transporting a compound useful in the present invention in or to a patient so that it can perform its intended function. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation, including the compound useful in the present invention, and must not cause harm to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; Cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethyleneglycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; surface-active agents; alginic acid; pyrogen-free water; isotonic saline solution; Ringer's solution; ethyl alcohol; phosphate buffer solutions and other non-toxic compatible substances used in pharmaceutical formulations.

A pharmaceutically acceptable carrier as used herein also includes any possible coating agents, antibacterial, antifungal, and absorption delaying agents, and the like, which are compatible with the activity of a compound useful in the present invention and are physiologically acceptable to the patient. Additional active compounds may also be included in the compositions. A pharmaceutically acceptable carrier may further include a pharmaceutically acceptable salt of a compound useful in the present invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the present invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, Pa.), which is incorporated herein by reference.

A pharmaceutically acceptable excipient refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance added to a pharmacological composition or otherwise used as a carrier medium, vehicle, or diluent to facilitate administration of the agent and is compatible with it. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

Delivery forms of pharmaceutical compositions containing one or more dosage units of active agents can be prepared using suitable pharmaceutical excipients and formulation techniques known or which may become available to those skilled in the art. The compositions can be administered according to the methods of the present invention by a suitable route of delivery, for example, oral, parenteral, rectal, topical or ocular routes, or by inhalation.

The preparation may be in the form of tablets, capsules, sachets, dragees, powders, granules, lozenges, powders for dilution, liquid preparations or suppositories. Preferably, the compositions are formulated for intravenous administration, topical administration or oral administration.

For oral administration, the compounds of the present invention may be provided in the form of tablets or capsules, or as a solution, emulsion, or suspension. For the manufacture of compositions for oral use, the compounds may be formulated to provide a dosage of, for example, from about 0.05 to about 100 mg/kg per day, or from about 0.05 to about 35 mg/kg per day, or from about 0.1 to about 10 mg/kg per day. For example, a total daily dosage of about 5 mg to 5 g per day may be achieved by administering a dose once, twice, three times, or four times per day.

Tablets for oral use may comprise a compound of the present invention mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrants, binders, lubricants, sweeteners, flavoring agents, coloring agents, and preservatives. Suitable inert fillers include sodium and calcium carbonate, sodium and calcium phosphate, lactose, starch, sugar, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and the like. Illustrative liquid excipients for oral use include ethanol, glycerol, water, and the like. Suitable disintegrants are starch, polyvinylpyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose, and alginic acid. Binders may include starch and gelatin. Lubricants, if present, may be magnesium stearate, stearic acid, or talc. If desired, tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption from the gastrointestinal tract, or may be enteric coated.

Capsules for oral administration include hard and soft gelatin capsules. For the manufacture of hard gelatin capsules, the compounds of the present invention can be mixed with a solid, semi-solid or liquid diluent. Soft gelatin capsules can be prepared by mixing a compound of the present invention with water, an oil such as peanut oil or olive oil, liquid paraffin, a mixture of mono- and diglycerides of short-chain fatty acids, polyethylene glycol 400 or propylene glycol.

Liquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups, or may be lyophilized or presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid compositions may optionally contain pharmaceutically acceptable excipients such as suspending agents (e.g. sorbitol, methylcellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, and the like); non-aqueous carrier vehicles such as oil (e.g. almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol or water; preservatives (e.g. methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents such as lecithin; and, if desired, flavoring or coloring agents.

The active agents of the present invention may also be administered by routes other than oral. For example, the compositions may be formulated for rectal administration as a suppository. For parenteral administration, including intravenous, intramuscular, intraperitoneal or subcutaneous routes, the compounds of the present invention may be provided in sterile aqueous solutions or suspensions buffered to a suitable pH and isotonicity, or in a parenterally acceptable oil. Suitable aqueous carrier media include Ringer's solution and isotonic sodium chloride. Such forms will be in unit-dose form such as ampoules or disposable administration devices, in multi-dose form such as vials from which a suitable dose can be withdrawn, or in solid form or preformulated concentrate form which can be used to prepare an injectable formulation. Illustrative dosages for administration by infusion may range from about 1 to 1000 mcg/kg/minute of the compound mixed with a pharmaceutical carrier over a period ranging from several minutes to several days.

For topical administration, the compounds may be mixed with a pharmaceutical carrier at a concentration of from about 0.1% to about 10% of the drug relative to the carrier medium. Another method of administering the compounds of the present invention may be a patch formulation for transdermal delivery.

Alternatively, in the methods of the present invention, the compounds of the present invention may be administered by inhalation, intranasal or oral routes, for example, via a spray formulation also containing a suitable carrier.

Methods of application.

The disclosed compounds are useful in the prevention or treatment of an HBV infection or a disease caused by HBV in a mammal in need thereof, more particularly in a human in need thereof.

In a non-limiting aspect, such compounds may (i) alter or disrupt HBV assembly and other functions of the HBV core protein required for HBV replication or the formation of infectious particles, (ii) inhibit the production of infectious viral particles or infection, or (iii) interact with the HBV capsid to cause the formation of defective viral particles with reduced infectivity or replication capacity, thereby acting as modulators of capsid assembly. In particular, and without being bound by any particular mechanism of action, the disclosed compounds are believed to be useful in the treatment of HBV by disrupting, accelerating, reducing, slowing down, and/or inhibiting normal viral capsid assembly and/or disassembly of immature or mature particles, thereby causing an aberrant capsid assembly resulting in antiviral effects such as disruption of virion assembly and/or disassembly, virion maturation, viral egress, and/or infection of target cells. The disclosed compounds may act as a capsid disruptor, interacting with mature or immature viral capsids to disrupt capsid stability, thereby affecting its assembly and/or disassembly. The disclosed compounds may disrupt protein folding and/or salt bridges required for stability, function, and/or normal

- 16 047929 viral capsid structure, thereby disrupting and/or accelerating capsid assembly and/or disassembly. The disclosed compounds can bind the capsid and alter the metabolism of cellular polyproteins and precursor substances, resulting in abnormal accumulation of protein monomers and/or oligomers and/or abnormal particles, which causes cellular toxicity and death of infected cells. The disclosed compounds can cause cessation of formation of capsids with optimal stability, affecting efficient uncoating and/or disassembly of viruses (e.g., during infectivity). The disclosed compounds can disrupt and/or accelerate capsid assembly and/or disassembly if the capsid protein is immature. The disclosed compounds can disrupt and/or accelerate capsid assembly and/or disassembly if the capsid protein is mature. The disclosed compounds can disrupt and/or accelerate capsid assembly and/or disassembly during viral infectivity, which can further attenuate HBV viral infectivity and/or reduce viral load. Disruption, acceleration, inhibition, slowing down and/or reduction of capsid assembly and/or disassembly by the disclosed compounds can remove the virus from the host. Removal of HBV from a subject by the disclosed compounds advantageously eliminates the need for chronic long-term therapy and/or reduces the duration of long-term therapy.

A further embodiment of the present invention is a method of treating a subject suffering from an infection caused by HBV, comprising administering to a subject in need of such treatment an effective amount of at least one compound of formula (I).

In another aspect, provided herein is a method for reducing the viral load associated with an HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the risk of recurrence of HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In addition, HBV serves as a helper virus for hepatitis delta virus (HDV), and it is estimated that more than 15 million people worldwide may be co-infected with HBV/HDV, which increases the risk of rapid progression to cirrhosis and increases liver failure compared to patients with HBV alone (Hughes, S.A. et al. Lancet 2011, 378, 73-85). Therefore, HDV infects subjects suffering from HBV infection. In a particular embodiment, the compounds of the present invention can be used in the treatment and/or prevention of HBV/HDV co-infection or diseases associated with HBV/HDV co-infection. Therefore, in a particular embodiment, the HBV infection is in particular an HBV/HDV co-infection and the mammal, in particular a human, may be co-infected with HBV/HDV or at risk of being co-infected with HBV/HDV.

In another aspect, provided herein is a method for suppressing or reducing the formation or level of presence of HBV DNA-containing particles or HBV RNA-containing particles in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the adverse physiological effects of HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for inducing remission of liver damage caused by HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the physiological impact of long-term antiviral therapy for HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for prophylactically treating an HBV infection in an individual in need thereof, wherein the individual is afflicted with a latent HBV infection, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In embodiments, the disclosed compounds are suitable for monotherapy. In embodiments, the disclosed compounds are effective against natural or native strains of HBV. In embodiments, the disclosed compounds are effective against strains of HBV that are resistant to currently known drugs.

In another embodiment, the compounds provided herein can be used in methods of modulating (e.g., inhibiting or disrupting) the activity of a

- 17 047929 HBV cccDNA plicity, function and viral replication.

In another embodiment, the compounds of the present invention can be used in methods for reducing or preventing the formation of HBV cccDNA.

In another embodiment, the compounds provided herein can be used in methods of modulating (e.g., inhibiting or disrupting) HBV cccDNA activity.

In another embodiment, the compounds of the present invention can be used in methods for reducing the formation of HBV cccDNA.

In another embodiment, the disclosed compounds can be used in methods of modulating, inhibiting, or disrupting the formation or release of HBV RNA particles from an infected cell.

In a further embodiment, the total load (or concentration) of HBV RNA particles is modulated. In a preferred embodiment, the total load of HBV RNA is reduced.

In another embodiment, the methods provided herein reduce the viral load in an individual to a greater extent or at a faster rate than administration of a compound selected from the group consisting of an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a viral maturation inhibitor, various capsid assembly modulators, antiviral compounds of various or unknown mechanisms, and any combination thereof.

In another embodiment, the methods provided herein cause a lower rate of viral mutation and/or viral resistance than administration of a compound selected from the group consisting of an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a viral maturation inhibitor, various capsid assembly modulators, antiviral compounds of various or unknown mechanisms, and any combination thereof.

In another embodiment, the methods provided herein further comprise administering to the individual at least one HBV vaccine, HBV nucleoside inhibitor, interferon, or any combination thereof.

In one aspect, provided herein is a method of treating an HBV infection in an individual in need thereof, comprising reducing the HBV viral load by administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, alone or in combination with a reverse transcriptase inhibitor, and then administering to the individual a therapeutically effective amount of an HBV vaccine.

A further embodiment of the present invention is a method of treating a subject suffering from an infection caused by HBV, comprising administering to a subject in need of such treatment an effective amount of at least one compound of formula (I).

In another aspect, provided herein is a method for reducing the viral load associated with an HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the risk of recurrence of HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for suppressing or reducing the formation or level of presence of HBV DNA-containing particles or HBV RNA-containing particles in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the adverse physiological effects of HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for inducing remission of liver damage caused by HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for reducing the physiological impact of long-term antiviral therapy for HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for prophylactically treating an HBV infection in an individual in need thereof, wherein the individual is afflicted with a latent HBV infection, comprising administering to the individual a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In one embodiment, the methods provided herein further

- 18 047929 include monitoring the HBV virus load in a subject, wherein the method is carried out over a period of time until the HBV virus is no longer detectable.

Combinations.

Provided herein are combinations of one or more of the disclosed compounds with at least one additional therapeutic agent. In embodiments, the methods provided herein may further comprise administering to the individual at least one additional therapeutic agent. In embodiments, the disclosed compounds are suitable for use in combination therapy. The compounds of the present invention may be useful in combination with one or more additional compounds useful for treating an infection caused by HBV. Such additional compounds may include compounds of the present invention or compounds known to be useful for treating, preventing, or reducing the symptoms or effects of an infection caused by HBV.

In an exemplary embodiment, the additional active ingredients are ingredients that are known or found to be effective in treating conditions or disorders involved in HBV infection, such as another modulator of HBV capsid assembly or a compound having activity against another target associated with a particular condition or disorder involved in HBV infection or the HBV infection itself. The combination may serve to increase the efficacy (e.g., by including in the combination a compound that enhances the activity or effectiveness of an active agent of the present invention), reduce one or more side effects, or reduce the required dose of an active agent of the present invention. In a further embodiment, the methods provided herein allow for the administration of at least one additional therapeutic agent at a lower dose or frequency compared to the administration of at least one additional therapeutic agent alone required to achieve a similar result in the prophylactic treatment of HBV infection in an individual in need thereof.

Such compounds include, but are not limited to, combination therapies for the treatment of HBV, HBV vaccines, HBV DNA polymerase inhibitors, immunomodulatory agents, toll-like receptor (TLR) modulators, interferon-alpha receptor ligands, hyaluronidase inhibitors, hepatitis b surface antigen (HBsAg) inhibitors, cytotoxic T lymphocyte-associated protein 4 (ipi4) inhibitors, cyclophilin inhibitors, HBV viral entry inhibitors, antisense oligonucleotide targeting viral mRNA, short interfering RNA (siRNA) and ddRNAi endonuclease modulators, ribonucleotide reductase inhibitors, HBV E antigen inhibitors, covalently closed circular DNA (cccDNA) inhibitors, farnesoid X receptor agonists, HBV antibodies, CCR2 chemokine antagonists, thymosin agonists, cytokines, nucleoprotein modulators, retinoic acid-inducible gene 1 stimulators, NOD2 stimulators, phosphatidylinositol 3-kinase (PI3K) inhibitors, indoleamine 2,3-dioxygenase (IDO) pathway inhibitors, PD-1 inhibitors, PD-L1 inhibitors, recombinant thymosin alpha-1, Bruton tyrosine kinase (BTK) inhibitors, KDM inhibitors, HBV replication inhibitors, arginase inhibitors and any other agents that interfere with the HBV life cycle and/or affect the outcome of HBV infection, or a combination thereof.

In embodiments, the compounds of the present invention may be used in combination with an HBV polymerase inhibitor, an immunomodulatory agent, an interferon such as pegylated interferon, a viral entry inhibitor, a viral maturation inhibitor, a capsid assembly modulator, a reverse transcriptase inhibitor, a cyclophilin/TNF inhibitor, an immunomodulatory agent such as a TLR agonist, an HBV vaccine, and any other agent that affects the life cycle of HBV and/or affects the consequences of HBV infection, or a combination thereof. In particular, the compounds of the present invention can be used in combination with one or more agents (or a salt thereof) selected from the group consisting of HBV reverse transcriptase inhibitors and DNA and RNA polymerase inhibitors, including but not limited to lamivudine (3TC, Zeffix, Heptavir, Epivir and Epivir-HBV), entecavir (Baraclude, Entavir), adefovir dipivoxil (Gepsera, Preveon, bis-POM PMEA), tenofovir disoproxil fumarate (Viread, TDF or PMPA);

interferons, including but not limited to interferon alpha (IFN-α), interferon beta (IFN-β), interferon lambda (IFN-λ), and interferon gamma (IFN-γ);

inhibitors of viral penetration;

inhibitors of viral maturation;

modulators of capsid assembly described in the literature, such as but not limited to BAY 41-4109; reverse transcriptase inhibitors;

an immunomodulatory agent such as a TLR agonist; and

- 19 047929 agents with a different or unknown mechanism of action, such as, but not limited to, AT-61 ((E)H-(1-chloro-3-oxophenyl-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide), AT-130 ((B)-K-(1-bromo-1-(2-methoxyphenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)-4-nitrobenzamide) and similar analogues.

In embodiments, the additional therapeutic agent is an interferon. The term interferon or IFN refers to any member of a family of highly homologous species-specific proteins that inhibit viral replication and cellular proliferation and modulate the immune response. Human interferons are grouped into three classes: type I, which includes interferon alpha (IFN-α), interferon beta (IFN-β), and interferon omega (IFN-ω), type II, which includes interferon gamma (IFN-γ), and type III, which includes interferon lambda (IFN-λ). Recombinant forms of interferons that have been developed and are commercially available are encompassed by the term interferon as used herein. Subtypes of interferons, such as chemically modified or mutated interferons, are also encompassed by the term interferon as used herein. Chemically modified interferons include pegylated interferons and glycosylated interferons. Examples of interferons also include, but are not limited to, interferon alpha-2a, interferon alpha-2b, interferon alpha-nl, interferon beta-1a, interferon beta-lb, interferon lambda-1, interferon lambda-2, and interferon lambda-3. Examples of pegylated interferons include pegylated interferon alpha-2a and pegylated interferon alpha-2b.

Accordingly, in one embodiment, the compounds of formula I can be administered in combination with an interferon selected from the group consisting of interferon alpha (IFN-α), interferon beta (IFN-β), interferon lambda (IFN-λ), and interferon gamma (IFN-γ). In one particular embodiment, the interferon is interferon alpha-2a, interferon alpha-2b, or interferon alpha-nl. In another particular embodiment, the interferon alpha-2a or interferon alpha-2b is pegylated. In a preferred embodiment, the interferon alpha-2a is pegylated interferon alpha-2a (PEGASYS).

In another embodiment, the additional therapeutic agent is selected from immunomodulatory or immunostimulatory therapies, which include biological agents belonging to the class of interferons.

Additionally, the adjunctive therapeutic agent may be an agent that disrupts the function of another essential viral protein(s) or host protein(s) required for HBV replication or survival.

In another embodiment, the additional therapeutic agent is an antiviral agent that blocks viral entry or maturation or that targets the HBV polymerase, such as nucleoside or nucleotide polymerase or non-nucleoside (nucleotide) polymerase inhibitors. In a further embodiment of the combination therapy, the reverse transcriptase inhibitor and/or DNA and/or RNA polymerase inhibitor is Zidovudine, Didanosine, Zalcitabine, ddA, Stavudine, Lamivudine, Abcavir, Emtricitabine, Entecavir, Apricitabine, Atevirapine, Ribavirin, Acyclovir, Famciclovir, Valaciclovir, Ganciclovir, Valganciclovir, Tenofovir, Adefovir, PMPA, Cidofovir, Efavirenz, Nevirapine, Delavirdine or Etravirine.

In one embodiment, the additional therapeutic agent is an immunomodulatory agent that induces a natural, limited immune response that results in immune responses to unrelated viruses. In other words, the immunomodulatory agent can affect the maturation of antigen-presenting cells, the proliferation of T cells, and the release of cytokines (e.g., IL-12, IL-18, IFN-alpha, -beta, and -gamma, and TNFalpha, among others).

In a further embodiment, the additional therapeutic agent is a TLR modulator or a TLR agonist, such as a TLR-7 agonist or a TLR-9 agonist. In a further embodiment of the combination therapy, the TLR-7 agonist is selected from the group consisting of SM360320 (9-benzyl-8-hydroxy-2-(2-methoxyethoxy)adenine) and AZD 8848 (methyl [3-({[3-(6-amino-2-butoxy-8-oxo-7,8-dihydro-9H-purin-9-yl)propyl][3-(4-morpholinyl)propyl]amino}methyl)phenyl]acetate).

In any of the methods provided herein, the method may further comprise administering to the individual at least one HBV vaccine, HBV nucleoside inhibitor, interferon, or any combination thereof. In one embodiment, the HBV vaccine is at least one of RECOMBIVAX HB, ENGERIX-B, ELOVAC B, GENEVAC-B, or SHANVAC B.

In another aspect, provided herein is a method of treating an HBV infection in an individual in need thereof, comprising reducing the HBV viral load by administering to the individual a therapeutically effective amount of a compound of the present invention alone or in combination with a reverse transcriptase inhibitor and then administering to the individual the

- 20 047929 therapeutically effective amount of HBV vaccine. The reverse transcriptase inhibitor may be one of Zidovudine, Didanosine, Zalcitabine, ddA, Stavudine, Lamivudine, Abacavir, Emtricitabine, Entecavir, Apricitabine, Atevirapine, Ribavirin, Acyclovir, Famciclovir, Valaciclovir, Ganciclovir, Valganciclovir, Tenofovir, Adefovir, PMPA, Cidofovir, Efavirenz, Nevirapine, Delavirdine or Etravirine.

In the case of any combination therapy described herein, the synergistic effect can be calculated, for example, using suitable methods such as the sigmoidal curve-Emax equation (Holford & Scheiner, 19981, Clin. Pharmacokinet. 6: 429-453), the Loewe additivity equation (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol. 114: 313-326) and the median effect equation (Chou & Talalay, 1984, Adv. Enzyme Regul. 22: 27-55). Each equation mentioned above can be applied to the experimental data to obtain the corresponding curve for assessing the effect of the drug combination. The corresponding graphs associated with the equations mentioned above are the concentration-effect curve, the isobologram curve and the additivity index curve, respectively.

Methods.

Also noteworthy are the methods for preparing the compound of formula (I) described in this document.

The method comprises at least one step from steps a), b), c), d), e) and f):

a) carrying out a reaction of a compound of formula (II) with a compound of formula (III)

in the presence of NaBH3CN to form a compound of formula (IV)

where n is an integer 0 or 1;

G ¹ is dichlorophenyl;

G ² is hydrogen or C _1-4 alkyl;

G3 is hydrogen or C _1-4 alkyl;

G ⁴ is a 5-10 membered monocyclic or bicyclic aromatic ring optionally containing one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of hydrogen, halogens, CN, CF3, CF2H, CFH2, CF2CH3, C _1-4 alkyl, OC _1-4 alkyl, OCF3, OCF2H and C ₃₄ cycloalkyl;

b) reacting a compound of formula (V) with a compound of formula (VI)

in the presence of potassium iodide (KI) to form a compound of formula (VII)

- 21 047929

where n equals 0,

G ¹ is dichlorophenyl;

G ² is hydrogen or C _1-4 alkyl;

G ⁵ is hydrogen or C _1-4 alkyl;

G ⁶ is C _2-5 alkynyl,

5-10 membered monocyclic or bicyclic aromatic ring optionally containing one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of hydrogen, halogens, CN, CF3, _CF2H , _CFH2 , CF2CH3, _C1-4alkyl , _OC1-4alkyl , OCF3, OC'FH, _{C3-4cycloalkyl} and _SO2N ( _CH3 ) ₂ ;

c) carrying out the reaction of the compound of formula (VIII)

with 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) or trimethylaluminium (Al(CH3)3), optionally in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) to form a compound of formula (IX)

where R is hydrogen or

n is an integer 0 or 1;

G ¹ is dichlorophenyl or tert-butoxide;

G ² is hydrogen or C _1-4 alkyl;

G ⁷ is C _1-4 alkyl,

C _1-4 alkyl substituted with one or more F,

A 5- to 10-membered monocyclic or bicyclic aromatic ring optionally containing one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5- to 10-membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of hydrogen, halogens, CN, CF3, _CF2H , _CFH2 , CF2CH3, _C1-4alkyl , _OC1-4alkyl , OCF3, OC'FH, _{C3-4cycloalkyl} and _SO2N ( _CH3 ) ₂ ,

C _2-5 alkynyl;

G ⁸ is hydrogen or C _1-4 alkyl;

d) carrying out a reaction of the compound of formula (X)

with a compound of formula (XI)

- 22 047929

G ⁹

G ¹⁰ (XI) in the presence of a non-nucleophilic base, more specifically cesium carbonate (Cs ₂ CO ₃ ) or sodium hydride (NaH), to form a compound of formula (XII)

where n is an integer 0 or 1; G ¹ is dichlorophenyl;

G ² is hydrogen or C _1-4 alkyl;

G ⁹ is hydrogen or O^alkyl,

G ¹⁰ is a 5-10 membered monocyclic or bicyclic aromatic ring optionally containing one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of hydrogen, halogens, CN, CF3, _CF2H , _CFH2 , _CF2CH3 , C1-4alkyl, _OC1-4alkyl , OCF3, OCF2H, _{C3-4cycloalkyl} , C(=O)Ot-Bu and _SO2N ( _CH3 ) ₂ ;

L represents bromide, chloride, or iodide;

e) carrying out the reaction of the compound of formula (XIII)

with a strong acid, more particularly hydrochloric acid (HCl) or trifluoroacetic acid (TFA), to form a compound of formula (XIV)

where n is an integer 0 or 1;

G ² is hydrogen or methyl;

G ¹¹ is H,

C _1-4 alkyl,

C _1-4 alkyl substituted with one or more F,

C _1-4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of hydrogen, halogens, CN, CF3, _CF2H , _CFH2 , CF2CH3, C _1-4 alkyl, _OC1-4 alkyl, OCF3, OCF2H and C _3-4 cycloalkyl;

Q is CH2, CHF, CH(CH2OH), CH(CH2F), C=CH2 or CHNHSO2CH3;

R is _CH2 or O;

T represents C=O or C=S;

f) carrying out the reaction of the compound of formula (XIV)

- 23 047929

with a compound of formula (XV)

O R ¹ ^CI (XV) in the presence of a non-nucleophilic base, more specifically sodium carbonate (Na2CO3), triethylamine (Et ₃ N) or diisopropylethylamine (DIPEA), to form a compound of formula (XVI)

where R ¹ is dichlorophenyl, and where G ² , n, Q, R, T and G ¹¹ have been defined in step e), provided that if the method comprises step a, the method further comprises step c, if the method comprises step b, the method further comprises step c, and if the method comprises step e, the method further comprises step f. Definitions.

Below are definitions of various terms used to describe the present invention. These definitions apply to the terms as they are used in this specification and claims, unless otherwise limited in specific instances, individually or as part of a larger group.

Unless otherwise defined, all technical and scientific terms used herein generally have the same meanings as commonly understood by one of ordinary skill in the art. In general, the nomenclature and laboratory procedures used in cell culture, molecular genetics, organic chemistry, and peptide chemistry used herein are well known and commonly used in the art.

As used herein, the singular forms refer to one or more than one (i.e., at least one) grammatical entity indicated in the singular. For example, an element refers to one element or more than one element. In addition, the use of the term including, as well as other forms such as include, includes, and included, is not limiting.

As used in the specification and claims, the term "comprising" may include the modifications "consisting of" and "consisting essentially of." The terms "comprises," "includes," "featured," "has," "may," "contains," and variations thereof, as used herein, are intended to be non-limiting transitional phrases, terms, or words that require the presence of the named ingredients/steps and permit the presence of other ingredients/steps. However, such description shall be construed to also describe compositions or methods as consisting of and consisting essentially of the listed compounds, which permit the presence of only the listed compounds, along with any pharmaceutically acceptable carriers, and to the exclusion of other compounds. All ranges disclosed herein include the stated endpoint and may be independently combined (e.g., a range of 50 mg to 300 mg includes the 50 mg and 300 mg endpoints and all values in between). The endpoints of the ranges and any values disclosed herein are not limited to a precise range or value; they are sufficiently imprecise to include values that are approximately equal to such ranges and/or values.

As used herein, approximate wording may be used to modify any quantitative representation that may vary without thereby changing the essential function to which it relates. Accordingly, the meaning modified by a term or terms such as significantly may not be limited to the precise value stated in some instances. In at least some instances, approximate wording may be consistent with the accuracy of the instrument for measuring the value.

- 24 047929

The term "alkyl" refers to a straight or branched chain alkyl group having from 1 to 12 carbon atoms in the chain. Examples of alkyl groups include methyl (Me, which may also be structurally represented by the symbol /), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and groups that would be considered equivalent to any of the above examples by one of ordinary skill in the art and the teachings provided herein. The term "C ₁₄ alkyl" as used herein refers to a straight or branched chain alkyl group having from 1 to 4 carbon atoms in the chain. The term "C 14 alkyl" as used herein refers to a straight or branched chain alkyl group having from 1 to 6 carbon atoms in the chain.

The term cycloalkyl refers to a saturated or partially saturated, monocyclic, fused polycyclic, or spiro-polycyclic carbocycle containing from 3 to 12 ring atoms per carbocycle. Illustrative examples of cycloalkyl groups include the following groups as suitably linked moieties:

A monocyclic, bicyclic or tricyclic aromatic carbocycle is an aromatic ring system consisting of 1, 2 or 3 rings, wherein said ring system consists only of carbon atoms; the term aromatic is well known to a person skilled in the art and denotes cyclically conjugated systems consisting of 4n+2 electrons, in other words, of 6, 10, 14, etc. π-electrons (Huckel's rule).

Specific examples of monocyclic, bicyclic or tricyclic aromatic carbocycles are phenyl, naphthalenyl, anthracenyl.

The term phenyl is the following fragment:

The term heteroaryl refers to an aromatic monocyclic or aromatic bicyclic ring system containing 5 to 10 ring members and which contains carbon atoms and from 1 to 4 heteroatoms independently selected from the group consisting of N, O and S. Included within the term heteroaryl are aromatic rings of 5 or 6 members, wherein the ring is composed of carbon atoms and contains at least one member which is a heteroatom. Suitable heteroatoms include nitrogen, oxygen and sulfur. In the case of 5-membered rings, the heteroaryl ring preferably contains one member which is nitrogen, oxygen and sulfur, and further no more than 3 additional nitrogen atoms. In the case of 6-membered rings, the heteroaryl ring preferably contains from 1 to 4, for example tetrazo

lyl (e.g., ), more particularly 1 to 3 nitrogen atoms. In the case where a 6-membered ring contains 3 nitrogen atoms, not more than 2 nitrogen atoms are adjacent. Examples of heteroaryl groups include furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, indazolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothiadiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl and quinazolinyl. Unless otherwise noted, a heteroaryl is attached to its side group at any heteroatom or carbon atom that results in a stable structure.

One skilled in the art will recognize that the types of heteroaryl groups mentioned or illustrated above are not exhaustive and that additional types within the scope of such defined terms may also be selected.

The term cyano refers to the -CN group.

The terms halo or halogen represent chlorine, fluorine, bromine or iodine.

The term substituted means that the stated group or moiety bears one or more substituents. The term unsubstituted means that the stated group bears no substituents. The term optionally substituted means that the stated group is unsubstituted or substituted by one or more substituents. When the term substituted is used to describe a structural system, it is intended that the substitution occur at any position in the system that valence permits. In cases where a stated moiety or group is not explicitly stated to be optionally substituted or substituted by any stated substituent, it is intended that such moiety or group be unsubstituted.

The terms para, meta and ortho have the meanings understood in the state of the art. Thus, for example, a fully substituted phenyl group has substituents in both ortho positions (o),

- 25 047929 adjacent to the point of attachment of the phenyl ring, both meta positions (m) and one para position (p) opposite the point of attachment. For further clarification, the position of the substituents in the phenyl ring 2 different ortho positions will be designated as ortho and ortho' and 2 different meta positions as meta and meta', as illustrated below.

ortho meta para'^u^'ortho' meta'

When referring to substituents on a pyridyl group, the terms para, meta, and ortho refer to the location of the substituent relative to the point of attachment of the pyridyl ring. For example, the structure below is described as 3-pyridyl with substituent X in the ortho position, substituent X ² in the meta position, and substituent X ³ in the para position.

X ¹

Some of the quantitative expressions given in this document are not qualified by the expression approximately. It is understood that, whether the term approximately is explicitly used or not, each quantity given in this document is meant to refer to the actual value stated, and it is also meant to be an approximation to such stated value that would be reasonably expected based on the knowledge of a person of ordinary skill in the art, including equivalents and approximations arising from experimental and/or measurement conditions for such stated value. In all cases where a yield is given as a percentage, such yield refers to the mass of the substance for which the yield is given, relative to the maximum amount of the same substance that could be obtained under the specified stoichiometric conditions. Concentrations given as percentage values refer to mass ratios unless otherwise indicated.

The terms buffered solution or buffer solution are used interchangeably herein in accordance with their standard meaning. Buffered solutions are used to control the pH of a medium, and their selection, use, and function are known to those of ordinary skill in the art. See, for example, G. D. Considine, ed., Van Nostrand's Encyclopedia of Chemistry, p. 261, ^5th ed. (2005), which describes, among other things, buffer solutions and how the concentrations of the substances making up the buffer relate to the pH of the buffer. For example, a buffered solution is prepared by adding MgSO ₄ and NaHCO ₃ to a solution in a 10:1 w/w ratio to maintain the pH of the solution at approximately 7.5.

Any formula shown herein is intended to represent compounds having the structure depicted by the structural formula, as well as some variations or forms. In particular, compounds of any formula shown herein may have centers of asymmetry and, therefore, exist in different enantiomeric forms. All optical isomers of compounds of the general formula and mixtures thereof are considered within the scope of the formula. Thus, any formula shown herein is intended to represent a racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. In addition, certain structures may exist as geometric isomers (i.e., cis and trans isomers), as tautomers, or as atropisomers.

It should also be understood that compounds having the same molecular formula, but different nature or sequence of bonding of their atoms or arrangement of their atoms in space, are designated by the term isomers.

Stereoisomers that are not mirror images of each other are called diastereomers, and stereoisomers that are non-superimposable mirror images of each other are called enantiomers. If a compound contains an asymmetric center, such as one bound to four different groups, a pair of enantiomers may exist. An enantiomer may be characterized by the absolute configuration of its asymmetric center and described by the Cahn and Prelog sequence rules for R and S isomers, or by the side to which the molecule rotates the plane of polarized light and is designated as dextrorotatory or levorotatory (i.e., as (+)- or (-)-isomers, respectively). A chiral compound may exist as either a single enantiomer or a mixture of both. A mixture containing enantiomers in equal proportions is called a racemic mixture.

Tautomers refer to compounds that are interchangeable forms of a particular

- 26 047929 compound structure, and which differ in the arrangement of hydrogen atoms and electrons. Thus, two structures can be in equilibrium by moving π-electrons and an atom (usually H). For example, enols and ketones are tautomers, since they quickly convert into each other by treatment with acid or base. Another example of tautomerism is the ac- and nitro forms of phenylnitromethane, which are similarly formed by treatment with acid or base.

Tautomeric forms may be important in achieving optimal chemical reactivity and biological activity of the compound of interest.

The compounds of the present invention may possess one or more centers of asymmetry; such compounds may therefore be obtained as individual (R)- or (R)-stereoisomers or mixtures thereof.

Unless otherwise indicated, the description or name of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures thereof, racemic or otherwise. Methods for determining stereochemical configuration and separating stereoisomers are well known in the art.

Certain examples contain chemical structures that are depicted as the absolute enantiomer but are intended to denote enantiomerically pure material of unknown configuration. In such cases, (R*) or (S*) or (*R) or (*S) is used in the name to indicate that the absolute stereochemical configuration of the corresponding stereocenter is unknown. Thus, a compound designated as (R*) or (*R) refers to an enantiomerically pure compound with an absolute configuration of either (R) or (S). In cases where the absolute stereochemistry has been confirmed, the structure is named using (R) and (S).

The symbols and are used to denote the same spatial configuration in the chemical structures shown herein. Similarly, the symbols ^11111111111 and ........ ^... are used to denote the same spatial configuration in the chemical structures shown herein.

Additionally, any formula provided herein is intended to also refer to hydrates, solvates, and polymorphs of such compounds and mixtures thereof, even if such forms are not explicitly recited. Certain compounds of Formula (I) or pharmaceutically acceptable salts of the compounds of Formula (I) can be prepared as solvates. Solvates include solvates formed by reacting or complexing the compounds of the present invention with one or more solvents, either in solution or as a solid or crystalline form. In some embodiments, the solvent is water and the solvates are hydrates. Additionally, certain crystalline forms of the compounds of Formula (I) or pharmaceutically acceptable salts of the compounds of Formula (I) can be prepared as co-crystals. In certain embodiments of the present invention, the compounds of Formula (I) were prepared in crystalline form. In other embodiments, the crystalline forms of the compounds of Formula (I) were cubic in nature. In other embodiments, the pharmaceutically acceptable salts of the compounds of Formula (I) were prepared in crystalline form. In still other embodiments, the compounds of formula (I) are obtained as several polymorphic forms, as a mixture of crystalline forms, as a polymorphic form, or as an amorphous form. In other embodiments, the compounds of formula (I) are converted in solution between one or more crystalline forms and/or polymorphic forms.

Furthermore, it is intended that any formula provided herein also refers to hydrates, solvates, and polymorphs of such compounds and mixtures thereof, even if such forms are not explicitly recited. Certain compounds of formula (1a) or pharmaceutically acceptable salts of compounds of formula (1a) can be prepared as solvates. Solvates include solvates formed by reacting or complexing compounds of the present invention with one or more solvents, either in solution or as a solid or crystalline form. In some embodiments, the solvent is water and the solvates are hydrates. Furthermore, certain crystalline forms of compounds of formula (1a) or pharmaceutically acceptable salts of compounds of formula (1a) can be prepared as co-crystals. In some embodiments of the present invention, compounds of formula (1a) were prepared in crystalline form. In other embodiments, crystalline forms of compounds of formula (1a) were cubic in nature. In other embodiments, pharmaceutically acceptable salts of compounds of formula (1a) were prepared in crystalline form. In still other embodiments, the compounds of formula (1a) are obtained as one of several polymorphic forms, as a mixture of crystalline forms, as a polymorphic form, or as an amorphous form. In other embodiments, the compound of formula (1a) is converted in solution between one or more crystalline forms and/or polymorphic forms.

A reference to a compound in this document means a reference to any of: (a) the actual form of such compound indicated, and (b) any form of such compound in the environment in which the compound

- 27 047929 is considered in the naming. For example, reference herein to a compound such as RCOOH embraces reference to any of, for example, RC.OOH, .,,,,, R-COOH^.) and R-COO ^- (cojl). In this example, P-COOH^, refers to the solid compound as it may be in, for example, the form of a tablet or some other solid pharmaceutical composition or preparation; RCOOH(sol.) refers to the undissociated form of the compound in a solvent; and R-COO-c,,,, refers to the dissociated form of the compound in a solvent, such as the dissociated form of the compound in an aqueous medium, whether such dissociated form is derived from R-COOH, a salt thereof, or any other substance that results in the formation of R-COO ^- upon dissociation in the medium in question. In another example, an expression such as exposing a substance to a compound of formula R-COOH refers to exposing such substance to a form or forms of the compound R-COOH that exists or exists in the environment in which such exposure occurs. In another example, an expression such as reacting a substance with a compound of formula R-COOH refers to reacting (a) such substance in a chemically corresponding form or forms of such substance that exists or exists in the environment in which such reaction occurs with (b) a chemically corresponding form or forms of the compound R-COOH that exists or exists in the environment in which such reaction occurs. In this regard, if such a substance is, for example, in an aqueous environment, it is understood that the compound R-COOH is in the same environment and therefore the substance is exposed to a type of compound such as R-COOH^o^, and/or R-COO ^- H.,, _{UI I} . where the subscript (aq.) denotes aqueous in accordance with its traditional meaning in chemistry and biochemistry. The carboxylic acid functional group has been chosen for such examples of the name; this choice is not intended to be limiting, however, but serves merely as an illustration. It is understood that similar examples may be provided with respect to other functional groups, including, but not limited to, hydroxyl, basic ring members containing nitrogen, such as those found in amines, and any other group that reacts or transforms in a known manner in a medium that contains the compound. Such reactions and transformations include, but are not limited to, dissociation, association, tautomerism, solvolysis including hydrolysis, solvation including hydration, protonation, and deprotonation. Further examples in this regard are not provided herein, since such reactions and transformations in a particular medium are known to one skilled in the art.

In another example, a zwitterionic compound is encompassed herein by reference to a compound known to form a zwitterion, even though it is not explicitly mentioned in its zwitterionic form. Terms such as zwitterion, zwitterions and their synonyms, zwitterionic compound(s), are standard IUPAC approved names that are widely known and form part of the standard sets of defined scientific names. In this regard, the name zwitterion is assigned to name identification CHEBI:27369 in the Chemical Entities of Biological Interest (ChEBI) dictionary for molecular substances. As is generally known, a zwitterion or zwitterionic compound is a neutral compound that possesses nominal unit charges of opposite sign. Such compounds are sometimes referred to by the term internal salts. In other sources, such compounds are referred to as dipolar ions, although the latter term is considered incorrect in yet other sources. As a specific example, aminoethanoic acid (the amino acid glycine) has the formula H2NCH2COOH, and it exists in some media (in this case, neutral media) as a zwitterion 'H3NCH2COO ^- . Zwitterions, zwitterionic compounds, internal salts and dipolar ions according to the known and well-established meanings of such terms are within the scope of the present invention, as would in any case be apparent to one of ordinary skill in the art. Since it is not necessary to name every embodiment that would be known to one of ordinary skill in the art, none of the structures of zwitterionic compounds that are related to the compounds of the present invention are explicitly indicated herein. They are, however, part of the embodiments of the present invention. Further examples in this regard are not provided in this document since such interactions and transformations in a particular environment that result in various forms of a particular compound are known to any person skilled in the art.

It is intended that any formula recited herein also provides for unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have the structures depicted by the formulae recited herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that may be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine such as ² H, ³ H, C ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²⁵ I, respectively. Such isotopically labeled compounds are useful in metabolic studies (preferably with C), kinetic studies

- 28 047929 reactions (e.g. with deuterium (i.e. D or ² H) or tritium (i.e. T or ³ H)), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including tissue distribution assays of the drug or substrate, or in radiotherapy of patients. In particular, a compound labeled with ¹⁸ F or C may be particularly advantageous in studies performed using PET or SPECT. In addition, substitution with heavier isotopes, such as deuterium (i.e. ² H), may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dose requirements. The isotopically labeled compounds of the present invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the Schemes or Examples and the methods of preparation described below, by replacing the non-isotopically labeled reagent with a commonly available isotopically labeled reagent.

It is intended that when referring to any formula specified in this document, the choice of a particular passage from the list of possible types for a specified variable does not determine the choice of the same type for a variable appearing elsewhere. In other words, if a variable appears more than once, the choice of a type from the specified list is independent of the choice of a type for the same variable elsewhere in the formula, unless otherwise specified.

In accordance with the above guidelines for interpretation in assignment of meanings and nomenclature, it should be understood that an explicit reference to a class in this document includes, where chemically feasible and unless otherwise indicated, an independent reference to embodiments of such class and a reference to each of the possible embodiments of subclasses of the class explicitly mentioned.

As a first example of substituent terminology, if a substituent S1 is one of _S1 and _S2 , and a substituent ^S2 is one of S3 and S4, then such assignments pertain to embodiments of the present invention specified by the selection that ^S1 is _S1 , and ^S2 is S3; ^S1 is _S1 , and ^S2 is S·; S2 is S2, and ^S2 is S3; S2 is _S2 , and S4 is _S4 ; and the equivalents of each of such selections. The more concise terminology 'S1 is one of _S1 and _S2 , and S4 is one of _S3 and _S4 is used accordingly herein for the purpose of brevity, but not by way of limitation. The above first example of substituent terminology, which is indicated by general terms, is intended to illustrate the various assignments of meanings to the substituents described in this document. The above rule specified in this document for substituents extends, where applicable, to such members as ^R1 , ^R2 , ^R3 , ^R4 , ^R5 , ^G1 , ^G2 , ^G3 , ^G4 , ^G5 , ^G6 , ^G7 , ^G8 , ^G9 , ^G10 , ^G11 , n, L, R, T, Q, W, X, Y and Z and any other general substituent symbols used in this document.

Furthermore, when more than one value assignment is given for any member or substituent, embodiments of the present invention include the various groupings that can be made from these value assignments taken independently and their equivalents. As a second example of substituent terminology, if a substituent Example is described herein as one of _S1 , _S2, and _S3 , the listing pertains to embodiments of the present invention for which Example is _S1 ; Example is _S2 ; Example is _S3 ; Example is one of _S1 and _S2 ; _Example is one of _S1 and S3; _Example is one of S2 and _S3 ; _Example is one of _S1 , _{S2 ,} and _S3 ; and _Example is any equivalent of each of such selections. The more concise terminology '^example is one of _S1 , _S2 and _S3 respectively is used herein for the purpose of brevity but not as a limitation. The above second example of substituent terminology, which is indicated by general terms, is intended to illustrate the various assignments of substituent values described herein. The above rule specified in this document for substituents extends, where applicable, to such members as ^R1 , ^R2 , ^R3 , ^R4 , ^{R5 , G1} , ^G2 , ^G3 , ^G4 , ^G5 , ^G6 , G7, ^G8 , ^G9 , ^G10 , ^G11 , n, L, R, T, Q, W, X, Y and Z and any other general substituent symbols used herein.

The nomenclature Ci-j, where j>i, when used herein in reference to a class of substituents, is intended to refer to embodiments of the invention for which each number of carbon-containing members from i to j, including i and j, is independently achieved. As an example, the term C _1-4 refers independently to embodiments that contain one carbon-containing member (C ₁ ), embodiments that contain two carbon-containing members (C2), embodiments that contain three carbon-containing members (C3), and embodiments that contain four carbon-containing members (C4).

The term C _nm alkyl refers to an aliphatic chain, whether straight or branched, with a total number of carbon-containing members in the chain N that satisfies the condition n<N<m, with m>n. It is assumed that any substitution by two substituents, referred to

- 29 047929 mine in this document, covers different possibilities of attachment, if more than one such possibility is possible. For example, a reference to a substitution by substituents with two points of attachment -A-B-, where A^B, refers herein to such a substitution by substituents with two points of attachment, where A is attached to the first substituted member, and B is attached to the second substituted member, and it also refers to such a substitution by substituents with two points of attachment, where A is attached to the second substituted member, and B is attached to the first substituted member.

The present invention also includes pharmaceutically acceptable salts of the compounds of formula (I), preferably the compounds described above and the specific compounds exemplified herein, and methods of treatment using such salts.

The term pharmaceutically acceptable means approved or capable of being approved by a regulatory agency of the federal or state government or by a similar body in countries other than the United States, or that is listed in the United States Pharmacopeia or other generally recognized pharmacopeia for use in animals and, more particularly, in humans.

A pharmaceutically acceptable salt is intended to mean a salt of the free acid or base of the compounds represented by formula (I) that is nontoxic, biologically tolerable, or otherwise biologically suitable for administration to a subject. It should possess the desired pharmacological activity of the parent compound. See, in general, G.S. Paulekuhn et al., Trends in Active Pharmaceutical Ingredient Salt Selection based on Analysis of the Orange Book Database, J. Med. Chem., 2007, 50:6665-72, S.M. Berge, et al., Pharmaceutical Salts, J Pharm Sci., 1977, 66:1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., WileyVCH and VHCA, Zurich, 2002. Examples of pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of patients without undesirable toxicity, irritation, or allergic reaction. The compound of formula (I) may possess a sufficiently acidic group, a sufficiently basic group, or both types of functional groups, and accordingly react with a variety of inorganic or organic bases and inorganic and organic acids to form a pharmaceutically acceptable salt.

The term prodrug means a precursor of the designated compound that, upon administration to a subject, is converted into the compound in vivo by a chemical or physiological process such as solvolysis or enzymatic cleavage, or under physiological conditions (e.g., a prodrug is converted to a compound of formula (I) upon adjustment to physiological pH). A pharmaceutically acceptable prodrug is one that is nontoxic, biologically tolerable, and otherwise biologically suitable for administration to a subject. Illustrative procedures for selecting and preparing suitable prodrug derivatives are described, for example, in Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

A pharmaceutically active metabolite means a pharmacologically active product of the metabolism of a compound of formula (I) or a salt thereof in the body. Prodrugs and active metabolites of a compound can be determined using standard techniques known or available in the art. See, for example, Bertolini, et al., J Med Chem. 1997, 40, 2011-2016; Shan, et al., J Pharm Sci. 1997, 86 (7), 765-767; Bagshawe, Drug Dev Res. 1995, 34, 220-230; Bodor, Adv Drug Res. 1984, 13, 224-331; Bundgaard, Design of Drugs (Elsevier Press, 1985) and Larsen, Design and Application of Drugs, Drug Design and Development (Krogsgaard-Larsen, et al., eds., Harwood Academic Publishers, 1991).

As used herein, the term composition or pharmaceutical composition refers to a mixture of at least one compound provided herein with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. There are many techniques in the art for administering the compound, including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, inhalation, and topical administration.

As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, involved in carrying or transporting a compound provided herein in or to a patient so that it can perform its intended function. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation, including the compound provided herein, and must not cause harm to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as

- 30 047929 such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; surface-active agents; alginic acid; pyrogen-free water; isotonic saline solution; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances used in pharmaceutical formulations. A pharmaceutically acceptable carrier as used herein also includes any possible coating agents, antibacterial, antifungal, and absorption delaying agents, and the like, which are compatible with the activity of a compound provided herein and are physiologically acceptable to the patient. Additional active compounds may also be included in the compositions. A pharmaceutically acceptable carrier may further include a pharmaceutically acceptable salt of a compound provided herein. Other additional ingredients that may be included in the pharmaceutical compositions provided herein are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

The term stabilizer as used herein refers to polymers capable of providing chemical inhibition or prevention of degradation of a compound disclosed herein. Stabilizers are added to compound formulations to improve the chemical and physical stability of the compound.

The term tablet as used herein denotes an orally administrable, single dose solid dosage form which can be prepared by compressing a drug substance or a pharmaceutically acceptable salt thereof with suitable excipients (e.g. fillers, disintegrants, lubricants, glidants and/or surface-active agents) by conventional tabletting techniques. The tablet can be prepared using conventional granulation techniques, e.g. wet or dry granulation, optionally milling the granules, followed by compression and optionally coating. The tablet can also be prepared by spray drying.

As used herein, the term capsule refers to a solid dosage form that encloses a medicinal product within either a hard or soft dissolvable container or shell. The container or shell may be formed from gelatin, starch and/or other suitable substances.

As used herein, the terms effective amount, pharmaceutically effective amount, and therapeutically effective amount refer to a non-toxic but sufficient amount of the agent necessary to produce the desired biological result. Such a result may be a reduction or alleviation of the signs, symptoms, or causes of a disease, or any other desired change in a biological system. A suitable therapeutic amount in any particular case can be determined by one of ordinary skill in the art using routine experimentation.

The terms combination, therapeutic combination, pharmaceutical combination or combination product as used herein refer to a non-fixed combination or kit of parts for combined administration wherein two or more therapeutic agents can be administered independently, simultaneously or separately within time intervals, particularly if those time intervals allow the combination partners to exhibit a combined, such as synergistic, effect.

The term modulators includes both inhibitors and activators, where inhibitors refer to compounds that reduce, prevent, inactivate, desensitize, or down-regulate HBV assembly and other functions of the HBV core protein required for HBV replication or formation of infectious particles.

As used herein, the term "capsid assembly modulator" refers to a compound that disrupts or accelerates, or inhibits, or hinders, or slows down, or decreases, or alters the normal assembly of a capsid (e.g., during maturation) or the normal disassembly of a capsid (e.g., during the period of infectivity) or disrupts the stability of a capsid, thereby inducing the occurrence of an aberrant capsid structure and function. In one embodiment, a capsid assembly modulator accelerates the assembly or disassembly of a capsid, thereby inducing the formation of an abnormal capsid structure. In another embodiment, a capsid assembly modulator interacts with (e.g., binds at an active site, binds at an allosteric site, modifies and/or interferes with folding, etc.) a capsid major assembly (CA) protein, thereby disrupting the assembly or disassembly of a capsid. In another embodiment, the capsid assembly modulator causes a disruption in the structure or function of CA (e.g., the ability of CA to assemble, disassemble, bind to a substrate, fold into an appropriate

- 31 047929 changing conformation, etc.), which weakens the infectivity of the virus and/or kills the virus.

As used herein, the term treatment or treating is defined as the use or administration of a therapeutic agent, i.e., a compound of the present invention (alone or in combination with another pharmaceutical agent), to a patient, or the use or administration of a therapeutic agent to isolated tissue or a cell line obtained from a patient (e.g., for diagnostics or ex vivo use), which is characterized by the presence of an HBV infection, a symptom of an HBV infection, or the potential for developing an HBV infection, in order to cure, treat, alleviate, reduce, modify, eliminate, reduce the intensity of, ameliorate, or influence the HBV infection, the symptoms of an HBV infection, or the potential for developing an HBV infection. Such treatments may be specifically adapted or modified based on knowledge gained in the field of pharmacogenomics.

As used herein, the term prevent or warn means the absence of development of a disorder or disease if it has not occurred, or the absence of development of an additional disorder or disease if the disorder or disease has already occurred. It also means the ability to prevent some or all of the symptoms associated with the disorder or disease.

As used herein, the term patient, individual, or subject refers to a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as sheep, bovine, porcine, canine, feline, and mouse mammals. Preferably, the patient, subject, or individual is a human.

In the methods of treatment according to the present invention, an effective amount of a pharmaceutical agent according to the present invention is administered to a subject suffering from such a disease or diagnosed as having such a disease, disorder or condition. An effective amount means an amount or dose sufficient to produce the generally desired therapeutic or prophylactic benefit in patients in need of such treatment of the said disease, disorder or condition. Effective amounts or doses of the compounds of the present invention can be determined by standard methods, such as modeling, dose escalation studies or clinical trials, and by taking into account conventional factors, such as the route or route of administration or delivery of the drug, the pharmacokinetics of the compound, the severity and course of the disease, disorder or condition, previous or ongoing therapy that the subject is receiving, the subject's health status and response to drugs and the judgment of the attending physician. An exemplary dose is in the range of about 0.001 to about 200 mg of compound per kg of subject body weight per day, preferably about 0.05 to 100 mg/kg/day or about 1 to 35 mg/kg/day in a single or divided dosage unit (e.g., BID, TID, QID). For a 70-kilogram human, an illustrative range of a suitable dose amount is about 0.05 to about 7 g/day or about 0.2 to about 2.5 g/day.

An exemplary dose of the compound is from about 1 mg to about 2500 mg. In some embodiments, the dose of a compound of the present invention used in the compositions described herein is less than about 10,000 mg, or less than about 8000 mg, or less than about 6000 mg, or less than about 5000 mg, or less than about 3000 mg, or less than about 2000 mg, or less than about 1000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dose of the second compound (i.e., another HBV treatment agent) described herein is less than about 1000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all integers or fractions therebetween.

Once the disease, disorder or condition of the patient has improved, the dosage may be adjusted for preventive or maintenance treatment. For example, the dosage or frequency of administration, or both, may be reduced depending on symptoms to a level that is necessary to maintain the therapeutic or preventive effect. Of course, if the symptoms have been relieved to a suitable level, treatment may be stopped. Patients may, however, require treatment intermittently over a long period of time whenever symptoms reappear.

Infections caused by HBV that can be treated according to the disclosed methods include infections caused by HBV genotypes A, B, C and/or D. However, in one embodiment,

- 32 047929 HBV of any genotype can be treated using the disclosed methods (treatment of all genotypes). Genotyping of HBV can be performed using methods known in the art, for example HBV genotyping using INNO-LIPA®, Innogenetics N.V., Ghent, Belgium).

In order to assist the reader of the present invention, the description has been divided into several paragraphs or sections. Such divisions should not be considered as separating the substance of a paragraph or section from the substance of another paragraph or section. On the contrary, the present description covers all possible combinations of the various sections, paragraphs and sentences that may be considered.

Each of the relevant disclosures of all sources mentioned in this document is specifically incorporated by reference. The following examples are offered for illustration only and not limitation.

Examples

Illustrative compounds useful in the methods of the present invention will now be described with reference to illustrative synthetic schemes for their general preparation set forth below and specific examples set forth below. It will be appreciated by those skilled in the art that for the preparation of the various compounds herein, the starting materials can be suitably selected so that the desired substituents are ultimately reacted in the schemes with protected or unprotected groups, as appropriate, to yield the desired product. Alternatively, it may be necessary or desirable to use, in place of the ultimately desired substituent, a suitable group that will not participate in the reactions and replace it, if necessary, with the desired substituent. Unless otherwise specified, the variables are as defined above for formula (I) and formula (1a). The reactions can be carried out at a temperature from the melting point to the boiling point of the solvent, preferably from 0°C to the boiling point of the solvent. The reaction mixtures can be heated using conventional heating or heating with microwave irradiation. Reactions can also be carried out in hermetically sealed high-pressure vessels at temperatures above the normal boiling point of the solvent.

The compounds of formula (I) and formula (1a) can be converted into their corresponding salts using methods known to those skilled in the art. For example, an amine of formula (I) is treated with trifluoroacetic acid, HCl or citric acid in a solvent such as Et2O, _CH2Cl2 , THF, MeOH, chloroform or _isopropanol to provide the corresponding salt form. Alternatively, trifluoroacetic acid or formic acid salts are obtained by purification conditions using reversed-phase HPLC. Crystalline forms of pharmaceutically acceptable salts of the compounds of formula (I) and formula (1a) can be obtained in crystalline form by recrystallization from polar solvents (including mixtures of polar solvents and aqueous mixtures of polar solvents) or from non-polar solvents (including mixtures of non-polar solvents).

If the compounds according to the present invention have at least one chiral center, they may accordingly exist as enantiomers. If the compounds have two or more chiral centers, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are within the scope of the present invention.

Compounds that are present as a stereoisomeric mixture (meaning a mixture of two or more stereoisomers and includes enantiomers, diastereomers, and combinations thereof) are separated by SFC separation.

The compounds may be obtained as individual forms, such as individual enantiomers, by form-specific synthesis or by resolution. The compounds may alternatively be obtained as mixtures of different forms, such as racemic (1:1) or non-racemic (non-1:1) mixtures. If racemic and non-racemic mixtures of enantiomers are obtained, the individual enantiomers may be isolated using conventional separation methods known to those skilled in the art, such as chiral chromatography, recrystallization, diastereomeric salt formation, derivatization into diastereomeric adducts, bioconversion, or enzymatic conversion. If regioisomeric or diastereomeric mixtures are obtained, if applicable, the individual isomers may be separated using conventional methods, such as chromatography or crystallization.

1. General information.

Chemical names.

Chemical names were obtained using the chemical software: ACD/ChemSketch, and may preferably follow IUPAC rules.

General procedure for LCMS methods.

High performance liquid chromatography (HPLC) measurements were performed using an LC pump, a diode array detector (DAD) or a UV detector, and a column as specified in the respective methods. Additional detectors were included when necessary (see the method table below).

- 33 047929

The column flow was directed to a mass spectrometer (MS) equipped with an atmospheric pressure ionization source. It is within the skill of the art to set adjustable parameters (e.g., scan range, dwell time, etc.) to obtain ions that allow the nominal monoisotopic molecular weight (MW) of the compound to be determined. Data collection was performed using appropriate software.

Compounds are described by their experimental retention times (Rt) and ions. Unless otherwise stated in the data table, the molecular ion indicated corresponds to [M+H]+ (protonated molecule) and/or [MH] ^- (deprotonated molecule). In case the compound was not capable of direct ionization, the adduct type is indicated (i.e. [M+NH4] ⁺ , [M+HCOO] ^- etc.). All results were obtained with experimental errors that are usually associated with the method used.

Hereinafter in this document, SQD means single quadrupole detector, MSD means mass selective detector, RT means room temperature, BEH means bridged ethylsiloxane/silica hybrid, DAD means diode array detector, HSS means high strength silica, Q-Tof means quadrupole time-of-flight mass spectrometers, CLND means chemiluminescent nitrogen detector, ELSD means evaporative light scattering detector.

LCMS Methods.

Table 5

Code method a Device Column Mobile phase Gradient Stream T col. Time I analyzed for A Thermoscientific Ultimate Agilent: Poroshell A: 0.1% _HCO2H2O 98% A for 2 min. 1 18.4

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3000 DAD and Brucker NST ultra EC-C18 (4 mkm, 4.6 x 100 mm) water/ B: 0.05% HCOOH in _CH3CN to 0% A in 10 min., holding for 3.4 min., again to 98% Aza in 1.3 min., holding for 1.7 min. 30 IN Waters: Acquity® UPLC®- DAD, SQD and ELSD Waters: HSS T3 (1.8 mkm, 2.1 * 100 mm) A: 10 mM _CH3COONH4 in 95% _{H2O +} 5% _CH3CN B: _CH3CN From 100% A to 5% A in 2.10 minutes, to 0% A in 0.90 minutes, to 5% A in 0.5 minutes. 0,6 55 3.5 With Waters: Acquity® UPLC® DAD and SQD Waters: BEH C18 (1.7 mkm, 2.1*50 mm) A: 10 mM _CH3COONH4 in 95% _{H2O +} 5% _CH3CN B: _CH3CN From 95% A to 5% A in 1.3 min, hold for 0.7 min. 0,8 55 2 D Waters: Acquity® UPLC®- DAD and SQD Waters: BEH (1.8 mkm, 2.1 * 100 mm) A: 10 mM _CH3COONH4 in 95% _{H2O +} 5% _CH3CN B: _CH3CN From 100% to 5% A in 2.10 minutes, to 0% A in 0.90 minutes, to 5% A in 0.5 minutes. 0,6 55 3.5 E Waters: Acquity® UPLC®DAD and SQD Waters: BEH (1.8 mkm, 2.1 * 100 mm) A: 10 mM _CH3COONH4 in 95% _{H2O +} 5% _CH3CN , B: _CH3CN From 100% A to 5% A in 2.10 minutes, to 0% A in 0.90 minutes, to 5% A in 0.5 minutes. 0,6 55 3.5 F Agilent Poroshell 120 SB-C18 4.6x30 mm, 2.7 mkm with UHPLC Guard Infinity Lab Poroshell 120 SB-C18 4.6x5 mm, 2.7 mkm Acetonitrile: water (99:1%), 0.1% formic acid B - water (0.1% formic acid) 0.01 min - 99% B 1.5 min - 0% B 2.2 min - 0% B 2.21 min - 99% B 3 60 5.92

- 35 047929

G Agilent 1200 with DAD Column Sunfire C18 (5 mkm, 4.6x50 mm) Al: 0.02% ammonium acetate; A2: 0.1% TFA; mobile phase Bl: acetonitrile From 95% A1 or A2 and 5% B to 5% A1 or A2 and 95% B 1,5 6.5 N Agilent 1260 with VWD Column Dikma plus C18 (5 mkm, 4.6x30 mm) Al: 0.02% ammonium acetate; A2: 0.1% TFA; B: acetonitrile From 95% A1 or A2 and 5% B to 5% A1 or A2 and 95% B 1,5 4 I Waters: Acquity® UPLC® DAD and SQD Waters: BEH (1.8 mkm, 2.1 x 100 mm) A: 0.1% NH4HCO3 in 95% _H2O +5% _CH3CN B: _CH3CN From 100% A to 5% Aza 2.10 min., to 0% A in 0.9 min., to 5% A in 0.5 min. 0,6 55 3.5 J Waters: Acquity® UPLC® DAD and SQD Waters: BEH (1.8 mkm, 2.1 x 100 mm) A: 10 mM CH3COONH4 in 95% _H2O +5% _CH3CN B: _CH3CN From 100% A to 5% Aza 2.10 min., to 0% A in 0.9 min., to 5% A in 0.4 min. 0,6 55 3.5

(flow rate is expressed in ml/min; column temperature (T) in °C; analysis time in minutes).

General procedure for SFCMS methods.

The SFC measurements were performed using a supercritical fluid chromatography (SFC) analytical system equipped with a two-component pump for the delivery of carbon dioxide (CO2) and modifier, an automatic dispenser, a column thermostat, a diode array detector equipped with a high-pressure flow cell capable of withstanding up to 400 bar. When equipped with a mass spectrometer (MS), the column flow was directed to the (MS). It is within the skill of the art to set the adjustable parameters (e.g., scan range, dwell time, etc.) to obtain ions that allow the determination of the nominal monoisotopic molecular weight (MW) of the compound. Data collection was performed using appropriate software.

SFC-MS analytical methods (flow rate is expressed in ml/min; column temperature (T) in °C; analysis time in minutes, back pressure (BPR) in bar).

SFC methods.

- 36 047929

Table 6

Method code Column Mobile phase Gradient Stream T col. Analysis time BPR SFC_A Daicel Chiralpak® IC3 Column (3.0 µm, 150 x 4.6 mm) A: _CO2 B: EtOH+0.2% _iPrNH2 from 10% to 50% B in 6 min., holding 3.5 min. 2,5 9.5 40 130 SFCB Daicel Chiralpak® AD3 column (3.0 µm, 150x4.6 mm) A: CO2 _B : EUNiPrOH (5050) + 0.2% _iPrNH2 from 10% to 50% B in 6 min., holding 3.5 min. 2,5 40 9.5 130 SFCC Daicel Chiralpak® OJ3 column (3.0 µm, 150x4.6 mm) A: _CO2 from 10% to 50% B in 6 min., hold for 3.5 min. 2,5 40 9.5 130 B: EtOH+0.2% iPrNH ₂ SFC_D Daicel Chiralpak® AD-H Column (3.0 µm, 150 x 4.6 mm) A: _CO2 from 10% to 50% B in 6 min., holding 3.5 min. 2,5 40 9.5 PO B: iPrOH+0.2% iPrNH ₂ SFC_E Daicel Chiralpak® AD-H Column (3.0 µm, 150 x 4.6 mm) A: _CO2 B: EtOH+0.2% _iPrNH2 from 10% to 50% B in 6 min., hold for 3.5 min. 2,5 40 9.5 PO

(flow rate is expressed in ml/min; column temperature (T) in °C; analysis time in minutes).

NMR analysis.

1H NMR spectra were recorded on i) a Bruker DPX 400 MHz spectrometer, or ii) a Bruker Avance 400 MHz spectrometer, or c) a Bruker Avance III 400 MHz spectrometer, or d) a Bruker Avance 600 MHz spectrometer, or e) a Bruker Avance NEO 400 MHz spectrometer, or f) a Bruker model AVIII 400 spectrometer, g) a ZKNJ BIXI-1 300 MHz, Bruker Avance III 400 MHz, or h) a Bruker AVANCE Neo 400 MHz.

NMR spectra were recorded at ambient temperature unless otherwise stated. Data are reported as follows: chemical shift in parts per million (ppm) relative to TMS (δ=0 ppm) on the scale, integration, multiplicity (s singlet, d=doublet, t=triplet, q=quartet, quin=quintet, sext=sextet, sept=septet, m multiplicity, b=broad, or a combination thereof), coupling constant(s) J are reported in hertz (Hz).

MS Analysis.

Mass spectra were acquired on a Shimadzu LCMS-2020-MSD or Agilent 1200/G6110A MSD using electrospray ionization (ESI) in positive mode unless otherwise stated.

2. Abbreviations.

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Table 7

Microwave. Under the influence of microwave radiation ADDP 1,1'-(Azodicarbonyl)dipiperidine A1Me ₃ Trimethylaluminium Water. Water Atm. Atmosphere Sun ₂ O Di-/77/9C/77-butyldimethyl ether and carbonate VOS tert-Butyloxycarbonyl BODIPY Boron-dipyrromethene Ьг Wide SA Capsid assembly DAST (diethylamino)sulfur trifluoride DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DCE 1,2-Dichloroethane DCM Dichloromethane dd A doublet of doublets DEA Diethylamine DIPE Diisopropyl ether DIPEA/DIEA N,TV-Diisopropylethylamine DMF Dimethylformamide DMF-DMA Dimethylformamide-dimethylacetal DMSO Dimethyl sulfoxide DNA Deoxyribonucleic acid EDCI N-(3-dimethylaminopropyl)-L-ethyl carboimidine hydrochloride ESI Electrospray ionization Et ₃ N Triethylamine _Et2O Ether, diethyl ether EtOAc/EA Ethyl acetate EtOH Ethanol FCC Normal phase chromatography on silica gel

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G grams h. hour HBV Hepatitis B virus NOAS Acetic acid HPLC High Performance Liquid Chromatography Hz Hertz z- _PrNH2 Isopropylamine z-PrOH/IPA Isopropyl alcohol ΚΟ/Βιι Potassium tert-Butoxide LAH Lithium aluminum hydride LCMS Liquid chromatography with mass spectrometry LHMDS, LiHMDS Lithium bis(trimethylsilyl)amide M Molar m Multiplet MeCN/ACN Acetonitrile MeOH Methanol ΜΓ Milligram MHz Megahertz min. Minutes ml Milliliters mcl Microliters mmol Millimoles mcmol Micromoles MS Mass spectra MsCl Mesityl chloride mass/charge Mass to charge ratio N Normal NaOAc/AcONa Sodium acetate NMR Nuclear magnetic resonance o/p During the night PCR Polymerase chain reaction RE Petroleum ether RMRA 9-(2-Phosphonyl-methoxypropyl)adenine RRt Parts per million

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ppt Sediment Ru Pyridine RNA Ribonucleic acid Rt Retention time k.t. Room temperature S Singlet Saturation Rich SFC Supercritical fluid chromatography t Triplet T ₃ R Propanephosphonic acid anhydride TBAI Tetrabutylammonium iodide TBD 1,5,7-Triazabicyclo[4.4.0]dec-5-ene TEA Triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layer chromatography TLR Toll-like receptor TNF Tumor necrosis factor V or volumes Volume in milliliters of solvent per gram of substrate Δ Heating with reflux condenser

3. Procedures and analysis of compounds.

3.1 Synthesis of 6-membered rings.

3.1.1. Synthesis of intermediate compounds.

3.1.1.1. Synthesis of intermediate compound I3

^H 2 N=π

Pd/ _CH2N /o

------------>- .Ν'' / MeOH ί ^Ν // Ν

κ. τ., 18 4. ⁰

Intermediate compound I1.

Benzyl-[(1S)-1-(1H-1,2,4-triazol-3-yl)ethyl]carbamate

AND

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Benzyl [DO)-1-amino-1-sulfanylidenepropan-2-yl]carbamate (3.36 g, 12.1 mmol, 86% purity) was dissolved in EtOH (135 mL). Formic acid hydrazine (4.36 g, 72.7 mmol) and HgCl ₂ (4.28 g, 15.7 mmol) were added. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was diluted in EtOAc (200 mL), washed with NaHCO ₃ (sat., aq.), brine, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The combined aqueous layers were extracted with

- 40 047929 with EtOAc ( _5x200 mL). The combined organic extracts were dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The fraction was combined with the previously obtained residue. The residue was solubilized in EtOH (135 mL) and stirred at 80 °C for 16 h. The reaction mixture was concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 90:10) to give intermediate compound I1 (1.82 g, 58%) as a white solid.

Intermediate compound I2.

Benzyl {(1S)-1-[1 -(dimethylsulfamoyl)-1 H-1,2,4-triazol-3 -yl]ethyl} carbamate

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I1 (2.10 g, 8.08 mmol, 95% purity) was dissolved in DMF (53 mL). _K2CO3 (3.35 g, 24.3 mmol) and dimethylsulfamoyl chloride (1.04 mL, 9.70 mmol) were added. The reaction mixture was stirred at room temperature for 16 h and diluted with H2O (50 mL). _The aqueous layer was extracted with EtOAc (3 x 100 mL). _The combined organic layers were washed with brine (3 x 100 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 90:10) to give intermediate compound I2 (1.42 g, 47%, purity 94%).

Intermediate connection I3.

3-[(1S)-1 - Aminoethyl] -N, N -dimethyl-1 H-1,2,4-triazole-1-sulfonamide

H ₂ N ™ / ^N ^ ° : N ' Sf /

ABOUT \

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I2 (1.42 g, 3.76 mmol, 94% purity) was dissolved in MeOH (40 mL). The mixture was purged under argon and Pd/C (10% purity, 142 mg, 0.13 mmol) was added. The reaction mixture was purged with H2 (3 times) and stirred at room temperature for 18 h. The reaction mixture was filtered on Celite® and the filtrate was concentrated under reduced pressure to give intermediate I3 (830 mg), which was used as such in the next step.

3.1.1.2. Synthesis of intermediate compound I5.

Intermediate connection I4.

Ethyl 5-(3,4-dichlorobenzoyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

_CO2ET

A mixture of 5-tert-butyl 3-ethyl-6,7-dihydro-2H-pyrazolo[3,4-c]pyridine-3,5(4H)-dicarboxylate (17.8 g, 60.4 mmol) and TFA (46.2 mL, 604 mmol) in DCM (500 mL) was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in DCM. The solution was cooled to 0 °C. Et ₃ N (33.6 mL, 242 mmol) was added, followed by 3,4-dichlorobenzoyl chloride (13.3 g, 63.4 mmol) and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water (300 mL). The layers were separated and the organic phase was dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 90:10 to 0:100) to give intermediate compound I4 (16.2 g, 73%) as a white solid.

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Intermediate connection I5.

Ethyl 2-{2-[(tert-butoxycarbonyl)amino]ethyl}-5-(3,4-dichlorobenzoyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

A mixture of intermediate I4 (4.00 g, 10.9 mmol), _Cs2CO3 (5.31 g, 16.3 mmol) _, and 3-(boc-amino)propyl bromide (2.56 g, 11.4 mmol) in MeCN (150 mL) was stirred at room temperature overnight. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 90:10 to 0:100) to give intermediate I5 (3.0 g, 54%) as a clear oil.

3.1.1.3. Synthesis of intermediate compound I6.

5-tert-Butyl-3-ethyl-2-(2-bromoethyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate

A mixture of 5-tert-butyl 3-ethyl-6,7-dihydro-2H-pyrazolo[4,3]pyridine-3,5(4H)-dicarboxylate (1.25 g, 4.23 mmol), 1,2-dibromoethane (4.02 g, 21.2 mmol) _{and K2CO3 (} 1.76 g, 12.7 mmol) in MeCN (125 mL) was stirred at reflux overnight. The reaction mixture was filtered through decalite while still hot and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, heptane/EtOAc, gradient 95:5 to 0:100) to give intermediate I6 (1.7 g, 60%) as a clear oil.

3.1.1.4. Synthesis of intermediate compound I8

Intermediate connection I7.

Ethyl (6R)-5-(3,4-dichlorobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3carboxylate

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of 5-tert-butyl 3-ethyl-(6R)-6-methyl-2,4,6,7-tetrahydropyrazolo[4,3-c]pyridine-3,5-dicarboxylate (4.52 g, 14.6 mmol) in DCM (20 mL) was added HCl (4 N in 1,4-dioxane, 36.5 mL, 146 mmol). The reaction mixture was stirred at room temperature for 18 h and concentrated to dryness. The residue was diluted in DCM (150 mL) and Et ₃ N (9.16 mL, 65.7 mmol) was added. The solution was cooled to 0 °C and a solution of 3,4-dichlorobenzoyl chloride (3.21 g, 15.3 mmol) in DCM (30 mL) was added dropwise. The reaction mixture was stirred at this temperature for 2 h and diluted with DCM (750 mL). The organic phase was washed with water (2x500 mL), _NaHCO3 (sat., aq., 500 mL) and brine, dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/ _MeOH , gradient from 100:0 to 95:5) to give intermediate compound I7 (4.63 g, 83%).

Intermediate connection I8.

Ethyl (6R)-2-{2-[(tert-butoxycarbonyl)amino]ethyl}-5-(3,4-dichlorobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

- 42 047929

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I7 (4.85 g, 12.1 mmol) in MeCN (180 mL) and DMF (5 mL) were added _Cs2CO3 (5.92 g, 18.2 mmol) and a solution of 2-(tert-butoxycarbonylamino)ethyl methanesulfonate (3.48 g, 14.5 mmol) in MeCN (20 mL). The reaction mixture was stirred at room temperature for 3 days and diluted with EtOAc (100 mL) and _H2O (100 mL). The layers were separated. The aqueous layer was extracted with EtOAc (2x100 mL). The combined organic extracts were washed with brine, dried ( _{Na2SO4 )} , filtered and concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, cyclohexane/(EtOAc/EtOH, 3:1), gradient from 95:5 to 50:50) to give intermediate compound I8 (3.30 g, 52%).

3.1.1.5. Synthesis of intermediate compound I9.

Ethyl (6R)-2-(2-bromoethyl)-5-(3,4-dichlorobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3c]pyridine-3-carboxylate

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I7 (4.00 g, 10.5 mmol) in DMF (60 mL) were added _Cs2CO3 (6.82 g, 20.9 mmol) and 1,2-dibromoethane (4.51 mL, 52.3 mmol). The reaction mixture was stirred at 50 _° C for 1 h and diluted with _H2O (50 mL). The aqueous layer was extracted with EtOAc (3x70 mL). The combined organic extracts were washed with brine (3x60 mL), dried ( _{Na2SO4 )} , filtered and concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, cyclohexane/EtOAc, gradient from 9:1 to 1:1) to give intermediate compound I9 (2.90 g, 55%) as a white foam.

3.1.1.6. Synthesis of intermediate compound I60

160

To a solution of 4-bromo-3-chlorobenzoic acid (3.00 g, 12.7 mmol) in tetrahydrofuran (40 mL) were added lithium chloride (1.62 g, 38.2 mmol) and 2 M isopropyl magnesium chloride in tetrahydrofuran (20 mL, 40 mmol) at -78 °C under nitrogen atmosphere. After stirring at -78 °C for 10 min, the mixture was warmed to 0 °C and stirred at this temperature for 1 h. Then N,N-dimethylformamide (4.69 g, 64.2 mmol) was added. The reaction mixture was warmed to room temperature and stirred for 1.5 h. The reaction mixture was quenched with saturated aq. NH4Cl (10 mL) and extracted three times with ethyl acetate (10 mL). The aqueous layer was acidified with 1 N aqueous hydrochloride solution to pH=3. The aqueous layer was extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to give intermediate compound I57 (1.9 g, yield 73%, purity 90% according to ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, DMSO-d6) δ 13.68 (s, 1H), 10.38 (s, 1H), 8.04-7.96 (m, 3H).

To a solution of intermediate I57 (1.00 g, 4.88 mmol, 90% purity) in N-dimethylformamide (10 mL) was added 60 wt % sodium hydride in mineral oil (215 mg, 5.38 mmol). The mixture was stirred at room temperature for 30 min. Then iodomethane (760 mg, 5.35 mmol) was added. The mixture was stirred at room temperature for 2 h. The mixture was poured into 1 N aqueous hydrochloric acid (5 mL) and extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with aqueous sodium bicarbonate (10 mL) and brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to give intermediate compound I58 (880 mg, 82% yield, 90% purity by 1H NMR) as

- 43 047929 yellow oil. 1H NMR (400 MHz, DMSO-d6) δ 10.37 (s, 1H), 8.06-7.98 (m, 3H), 3.91 (s, 3H).

To a solution of intermediate I58 (250 mg, 1.13 mmol, 90% purity) in dichloromethane (10 ml) was added diethylaminosulfur trifluoride (920 mg, 5.71 mmol) at -78 °C. The mixture was stirred at room temperature overnight. The mixture was poured into water (10 ml) and extracted twice with dichloromethane (10 ml). The combined organic phases were washed with brine (5 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give intermediate I59 (170 mg, 61% yield, 90% purity by ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.10-8.09 (m, 1H), 8.04-8.02 (m, 1H), 7.76-7.74 (m, 1H), 6.97 (t, J=54.8 Hz, 1H), 3.96 (s, 3H).

To a solution of intermediate I59 (170 mg, 0.694 mmol, 90% purity) in methanol (3 mL) and tetrahydrofuran (3 mL) was added lithium hydroxide hydrate (45 mg, 1.1 mmol) in water (1.5 mL). The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give a residue. The residue was diluted with water (10 mL) and the resulting mixture was acidified with 1 N aqueous hydrochloride solution to pH = 3. The mixture was extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give intermediate I60 (150 mg, 94% yield, 90% purity by ¹ H NMR) as a white solid. ¹ H NMR (400 MHz, DMSO-d6) δ 13.62 (s, 1H), 8.04-8.02 (m, 2H), 7.85-7.83 (m, 1H), 7.28 (t, J=53.6 Hz, 1H).

3.1.1.7. Synthesis of intermediate compound I64

To a solution of methyl 4-chloro-3-methylbenzoate (2.00 g, 10.8 mmol) and N-bromosuccinimide (5.78 g, 32.5 mmol) in carbon tetrachloride (90 mL) was added 2,2'-azobis(2-methylpropionitrile) (450 mg, 2.74 mmol). The mixture was stirred at 80 °C overnight. The reaction mixture was then filtered and the filtrate was washed with water (10 mL), dried over anhydrous sodium sulfate and concentrated to give a residue which was triturated with n-hexane (10 mL) to give intermediate I61 (3.12 g, 80% yield, 95% purity by ¹ H NMR) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.49 (d, J=2.8 Hz, 1H), 7.96 (dd, J=11.2, 2.8 Hz, 1H), 7.70 (d, J=11.2 Hz, 1H), 7.54 (s, 1H), 3.93 (s, 3H).

To a solution of intermediate I61 (3.12 g, 8.66 mmol, 95% purity) in acetone (100 mL) and water (20 mL) was added silver nitrate(O) (2.94 g, 17.3 mmol) at room temperature. The mixture was heated under reflux under nitrogen for 1.5 h. The reaction mixture was then filtered through a pad of celite and the filtrate was concentrated to remove the organic solvent. The aqueous phase was extracted three times with dichloromethane (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to give intermediate I62 (1.7 g, 94% yield, 95% purity by 1H NMR) as a yellow oil. 1H NMR (400 MHz, CDCl ₃ ) δ 10.49 (s, 1H), 8.57 (d, J=2.4 Hz, 1H), 8.19 (dd, J=8.4, 2.0 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 3.95 (s, 3H).

To a solution of intermediate I62 (300 mg, 1.44 mmol, 95% purity) in dichloromethane (5 mL) was added diethylaminosulfur trifluoride (1.157 g, 7.175 mmol) at -78 °C. The mixture was stirred at room temperature overnight. The mixture was poured into water (10 mL) and extracted twice with dichloromethane (10 mL). The combined organic phases were washed with brine (5 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give intermediate II63 (270 mg, 81% yield, 95% purity by ¹ H NMR) as a yellow oil.

1H NMR (400 MHz, CDCl ₃ ) δ 8.39 (s, 1H), 8.13 (d, J=11.2 Hz, 1H), 7.57 (d, J=11.2 Hz, 1H) , 7.01 (t, J=72.8 Hz, 1H), 4.00 (s, 3H).

To a solution of intermediate I63 (270 mg, 1.16 mmol, 95% purity) in methanol (3 mL) and tetrahydrofuran (3 mL) was added lithium hydroxide hydrate (75 mg, 1.8 mmol) in water (1.5 mL). The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give a residue. The residue was diluted with water (10 mL) and the resulting mixture was acidified with

- 44 047929

1N aqueous hydrochloride solution to pH=3. The mixture was extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give intermediate compound I64 (220 mg, yield 87%, purity 95% according to ¹ H NMR) as a white solid. ¹ H NMR (400 MHz, DMSO-d6) δ 13.40 (s, 1H), 8.18-8.17 (m, 1H), 8.10-8.08 (m, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.28 (t, J=54.0 Hz, 1H).

3.1.1.8. Synthesis of intermediate compound I69

A mixture of methyl 3-bromo-4-methylbenzoate (2.00 g, 8.70 mmol), zinc cyanide (1.64 g, 14.0 mmol) and tetrakis(triphenylphosphine)palladium (1.0 g, 0.87 mmol) in N-dimethylformamide (30 mL) was stirred at 100 °C under nitrogen atmosphere for 16 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. Then, the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to give intermediate compound I65 (1.4 g, yield 92%, purity 90% according to ¹ H NMR) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.27 (d, J=1.6 Hz, 1H), 8.13 (dd, J=8.0, 1.6 Hz, 1H), 7.41 (d, J=7.6 Hz, 1H), 3.94 (s, 3H), 2.61 (s, 3H).

To a solution of intermediate I65 (500 mg, 2.86 mmol) in perchloromethane (8 mL) were added N-bromosuccinimide (1.27 g, 7.14 mmol) and benzoyl peroxide (75%, 28 mg, 0.086 mmol). The mixture was then stirred at 85 °C overnight. The mixture was diluted with water (20 mL) and extracted with dichloromethane (20 mL). The organic layer was washed with aqueous sodium bicarbonate (20 mL) and brine (20 mL), dried over Na ₂ SO ₄ , and concentrated to give crude intermediate I66 (900 mg, 95% yield, 80% purity by ¹ H NMR) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.32 (dd, J=8.0, 1.6 Hz, 1H), 8.27 (d, J=1.2 Hz, 1H), 8.12 (d, J=8.4 Hz, 1H), 7.00 (s, 1H), 3.98 (s, 3H).

To a solution of intermediate I66 (500 mg, 1.50 mmol) in acetonitrile (6 mL) was added a solution of silver nitrate (765 mg, 4.50 mmol) in water (3 mL). The mixture was then stirred at 80 °C for 6 h. The mixture was filtered and the filtrate was diluted with water (10 mL). The mixture was extracted with EtOAc (20 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 100:1 to 5:1) to give intermediate I67 (130 mg, yield 46%, purity 90% according to ¹ H NMR) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.42 (s, 1H), 8.48 (d, J=1.2 Hz, 1H), 8.40 (dd, J=8.4, 0.8 Hz, 1H), 8.13 (d, J=8.0 Hz, 1H), 4.01 (s, 3H).

To a solution of intermediate I67 (130 mg, 0.69 mmol) in dichloromethane (4 mL) was added (diethylamino)sulfur trifluoride (222 mg, 1.38 mmol) at 0 °C. The mixture was then stirred at 0 °C for 15 min. The reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (10 mL). The organic layer was washed with saturated aqueous sodium bicarbonate (10 mL) and brine (10 mL), dried over Na ₂ SO ₄ and concentrated to give intermediate I68 (140 mg, yield 97%, purity 90% by ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.42 (s, 1H), 8.37 (d, J=8.0 Hz, 1H), 7.86 (d, J=8.0 Hz, 1H), 6.97 (t, J=54.0 Hz, 1H), 3.99 (s, 3H).

To a solution of intermediate I68 (140 mg, 0.66 mmol) in tetrahydrofuran/water (2:1, 3 mL) was added lithium hydroxide hydrate (34 mg, 0.80 mmol). The mixture was stirred at room temperature for 2 h. Then the mixture was diluted with water (5 mL) and acidified with 1N hydrochloric acid to pH = 2. The mixture was extracted with ethyl acetate (10 mL). The organic phase was washed with brine (10 mL), dried over Na ₂ SO ₄ and concentrated to give intermediate I69 (100 mg, yield 76.6%, purity 95% by ¹ H NMR) as a pale yellow solid.

¹ H NMR (400 MHz, DMSO-d6) δ 13.87 (brs, 1H), 8.45 (s, 1H), 8.35 (d, J=8.4 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.35 (t, J=54.0 Hz, 1H).

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3.1.1.9. Synthesis of intermediate compound I71

OFF

(Diethylamino)sulfur trifluoride (532 mg, 3.30 mmol) was added to an ice-cold solution of methyl 3-fluoro-4-formyl benzoate (300 mg, 1.65 mmol) in dichloromethane (7 mL). The reaction mixture was stirred at 0 °C for 1 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (7 mL) and extracted with dichloromethane (10 mL). The organic layer was washed with brine (10 mL), dried over Na ₂ SO ₄ and concentrated to give intermediate I70 (300 mg, 89% yield, 90% purity by ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.92 (d, J=8.0 Hz, 1H), 7.79 (d, J=10.4 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H), 6.92 (t, J=54.4 Hz, 1H), 3.95 (s, 3H).

To a solution of intermediate I70 (100 mg, 0.49 mmol) in tetrahydrofuran/water (2:1, 3 ml) was added lithium hydroxide hydrate (23 mg, 0.54 mmol). The mixture was stirred at room temperature for 1 h. The mixture was then diluted with water (5 ml) and acidified with 0.5 N hydrochloric acid to pH= ₄ . The mixture was then extracted with ethyl acetate (10 ml). The organic phase was washed with brine (10 ml), dried over _Na2SO4 and concentrated to give intermediate I71 (84 mg, 90% yield, 95% purity by 1H NMR) as a pale yellow solid.

1H NMR (400 MHz, DMSO-d6) δ 13.47 (brs, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.82-7.76 (m, 2H), 7.29 (t, J=54.0 Hz, 1H).

3.1.1.10. Synthesis of intermediate compound I76

Υί nvg/asOn Υ η DIBALH (2.6 eq.) ^Ck yf4, DAST (5.6 eq.) YY

Y ^{N Cl} 110°C. 5 h. Y ^{N Br} THF, -78°C, 3 h. ^0? YYrDCM, -78°C ~ κ. τ„ Y ^{N Br} he he 3 h

1702-17-6 |72 I73I74

DMF/MeOH,90°C, 5 4.

I75I76

To a solution of 3,6-dichloropicolinic acid (9.5 g, 49.5 mmol) in acetic acid (19 mL) was added 33% hydrobromide in acetic acid (19 mL). The mixture was heated to 110 °C and stirred at 110 °C for 1 h. After adding additional 33% hydrobromide in acetic acid (10 mL), the mixture was stirred at 110 °C for an additional 2 h. Additional 33% hydrobromide in acetic acid (10 mL) was added, and then the mixture was stirred at 110 °C for an additional 2 h. After cooling to room temperature, the mixture was poured into ice water (100 mL). The white solid was collected by filtration, washed twice with water (10 mL), and dried under reduced pressure to give intermediate I72 (6.46 g, 90% purity by 1H NMR, 50% yield). 1H NMR (400 MHz, DMSO-d6) δ 14.15 (br s, 1H), 8.04 (d, J=8.4 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H).

To a solution of intermediate I72 (2 g, 90% purity, 7.61 mmol) in tetrahydrofuran (40 mL) was slowly added 1.5 M diisobutylaluminum hydride in toluene (13 mL, 19.5 mmol) at -60 °C under nitrogen atmosphere. After stirring at -60 °C for 3 h, the reaction mixture was quenched with saturated aqueous ammonium chloride solution (10 mL), diluted with water (40 mL) and ethyl acetate (40 mL). The organic layer was separated and the aqueous layer was extracted twice with ethyl acetate (40 mL). The combined organic layers were washed with brine (40 mL), dried over Na ₂ SO ₄ (solid), and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1) to give intermediate compound I73 (1.52 g, 90% purity by 1H NMR, 82% yield) as a yellow solid. 1H NMR (400 MHz, _CDCl3 ) δ 10.16 (s, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.63 (d, J=8.4 Hz, 1H).

To a solution of intermediate I73 (1.52 g, 90% purity, 6.21 mmol) in dichloromethane (40 mL) was added diethylaminosulfur trifluoride (5.6 g, 34.7 mmol) at -78 °C. After stirring at room temperature for 3 h, the mixture was poured into cold saturated aqueous sodium bicarbonate solution (100 mL) and extracted twice with ethyl acetate (50 mL). The combined organic layers were washed with brine (100 mL), dried over Na ₂ SO ₄ (solid), and filtered.

- 46 047929

The filtrate was concentrated in vacuo to give a residue which was purified by column chromatography on silica gel (petroleum ether:ethyl acetate=20:1) to give intermediate compound I74 (1.45 g, 90% purity according to ¹ H NMR, 87% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.65 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 6.81 (t, J=53.6 Hz, 1H).

A solution of intermediate I74 (1.5 g, 90% purity, 5.57 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (546 mg, 0.746 mmol) and triethylamine (1.26 g, 12.5 mmol) in methanol (6 ml) and N-dimethylformamide (6 ml) was stirred at 90°C under carbon monoxide atmosphere (0.5 MPa) for 5 h. After cooling to room temperature, the mixture was concentrated in vacuo to give a residue which was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to give intermediate I75 (1.1 g, 90% purity by ¹ H NMR, 80% yield) as a light yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) 8.19 (d, J=8.0 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 6.91 (t, J=53.6 Hz, 1H), 4.03 (s, 3H).

To a solution of intermediate I75 (200 mg, 0.85 mmol) in methanol (2 mL) was added 5 M aqueous lithium hydroxide solution (0.5 mL, 2.5 mmol) at room temperature. After stirring at 30 °C overnight, the reaction mixture was diluted with water (10 mL), acidified with 1 M aqueous hydrochloride solution to pH ~3 and then extracted three times with ethyl acetate (5 mL). The combined organic layers were washed with brine (10 mL), dried over Na ₂ SO ₄ (solid) and filtered. The filtrate was concentrated in vacuo to give intermediate I76 (180 mg, 95% purity by ¹ H NMR, 96% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.31 (d, J=8.4 Hz, 1H), 8.08 (d, J=8.4 Hz, 1H), 6.96 (t, J=53.6 Hz, 1H).

3.1.1.11. Synthesis of intermediate compound I79

Η

179

To a solution of sodium methoxide (25% in methanol) (20 mL, 87.5 mmol) in methanol (17 mL) at -10 °C, a solution of 3-fluoro-2-(trifluoromethyl)benzaldehyde (4.20 g, 21.9 mmol) and methyl 2-azidoacetate (8.5 mL, 87.5 mmol) in methanol (4.2 mL) was added using a syringe pump over 1 h (0.3 mL/min). The reaction mixture was stirred for 5 h while maintaining the temperature below 5 °C. Ice water was added to the reaction mixture and stirred for 10 min to ensure that the reaction mixture reached room temperature. The precipitate was filtered, washed with water and dried under vacuum to give methyl (7)-2-azido-3-(3-fluoro-2-(trifluoromethyl)phenyl)acrylate I77 (1.90 g, 30% yield) as a white solid.

The reaction was carried out in 9 reaction mixtures, for each of which the following procedure was followed. A solution of methyl (7)-2-azido-3-(3-fluoro-2-(trifluoromethyl)phenyl)acrylate I77 (210 mg, 0.726 mmol) in m-xylene (6 ml) was stirred on a hot plate preheated to 160°C for 1 h. The 9 reaction mixtures were combined and coevaporated with water (3 x 300 ml). The residue was purified by column purification (cyclohexane/EtOAc from 90/10 to 50/50). The obtained material was triturated in a mixture of EtOAc and cyclohexane. The precipitate was filtered to give methyl 5-fluoro-4-(trifluoromethyl)III-indole-2-carboxylate I78 (1.20 g, yield 70%) as a white solid.

To a solution of methyl 5-fluoro-4-(trifluoromethyl)-III-indole-2-carboxylate I78 (1.20 g, 4.60 mmol) in THF (10 mL) was added a solution of NaOH (551 mg, 13.8 mmol) in water (5 mL). The reaction mixture was stirred at room temperature for 46 h. The reaction mixture was diluted with DCM (100 mL) and water (50 mL) and the layers were separated. The aqueous layer was acidified with aqueous HCl (1 N) then extracted with DCM (3 x 100 mL) and MeOH (a few drops). The combined organic layers were dried over sodium sulfate, filtered and concentrated to dryness to give 5-fluoro-4-(trifluoromethyl)-III-indole-2-carboxylic acid I79 (1.05 g, 92% yield) as a white powder.

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3.1.1.12. Synthesis of intermediate compound I85

To a solution of methyl 4-bromo-5-fluoro-1H-indole-2-carboxylate (2.27 g, 8.34 mmol) in DCM (32 mL) and acetonitrile (18 mL) were added DMAP (1.33 g, 10.9 mmol) and di-tert-butyl dicarbonate (2.58 g, 11.8 mmol). The solution was stirred at room temperature for 1 h. The mixture was diluted with DCM (120 mL) and _H2O (50 mL). The layers were separated and the organic layer was washed with H2O (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromatography (cyclohexane/EtOAc, 100/0 to 90/10) to give 1(tert-butyl)-2-methyl-4-bromo-5-fluoro-1H-indole-1,2-dicarboxylate I80 (2.66 g, 86% yield) as a colourless oil.

I80 (2.66 g, 7.15 mmol) was dissolved in a mixture of 1,4-dioxane (53 mL) and water (10 mL). The mixture was degassed with argon for 20 min under vigorous stirring, followed by S-Phos ligand (293 mg, 0.715 mmol), potassium carbonate (2.96 g, 21.4 mmol), and potassium vinyl trifluoroborate (1.92 g, 14.3 mmol). The flask was purged with vacuum/argon three times, then palladium acetate (0.080 g, 0.357 mmol) was added. The flask was again purged with vacuum/argon 3 times, then stirred at 75°C for 1 h. The mixture was diluted with EtOAc (350 mL), then washed with _H2O (2x70 mL), brine (2x70 mL), dried over sodium sulfate, filtered, then concentrated in vacuo to give 1-(tert-butyl)-2-methyl-5-fluoro-4-vinyl-1H-indole-1,2-dicarboxylate I81 (2.55 g crude material, which was used as such in the next step).

To a solution of I81 (1.00 g, 3.13 mmol) in acetone (50 mL) and water (10 mL) were added osmium tetroxide (0.313 g, 0.094 mmol) and sodium periodate (3.35 g, 15.7 mmol). The reaction mixture was stirred at room temperature for 7 h. The reaction mixture was diluted with water and acetone, the suspension was filtered on a sintered glass filter and the residue was washed with acetone. The filtrate was concentrated in vacuo to remove most of the acetone. The resulting solution was diluted with EtOAc (200 mL), the layers were separated and the aqueous layer was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash chromatography (cyclohexane/EtOAc 100/0 to 95/5) to give 1-(tert-butyl)-2-methyl-5-fluoro-4-formyl-1H-indole-1,2-dicarboxylate I82 (623 mg, 62% yield) as a white powder.

In a microwave reaction vial, I82 (0.414 g, 1.29 mmol) was dissolved in DCM (5.8 mL). The solution was cooled to 0 °C, then DAST (0.51 mL, 3.87 mmol) was added dropwise. The reaction mixture was allowed to reach room temperature over 15 min, the vial was sealed, then stirred at 50 °C for 2.5 h and 16 h at room temperature. The reaction mixture was cooled to 0 °C, then quenched by slowly adding saturated aqueous NaHCO ₃ (5 mL), resulting in vigorous bubbling. The aqueous layer was extracted with DCM (3 x 15 mL). The combined organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by column chromatography (cyclohexane/EtOAc 100/0 to 95/5) to give 1-(tert-butyl)-2-methyl-4-(difluoromethyl)-5-fluoro-1H-indole-1,2-dicarboxylate I83 (394 mg, 84%) as a yellow oil.

To a solution of I83 (597 mg, 1.74 mmol) in DCM (15 mL) was added HCl (6.52 mL, 1 M in dioxane, 26.1 mmol). The reaction mixture was stirred at room temperature for 20 h. Incomplete conversion by LCMS. The solution was heated to 40 °C for 6 h. Incomplete conversion was still observed by LCMS. The reaction mixture was concentrated in vacuo, then diluted in DCM (7 mL), and B (3.00 mL, 12 mmol) was added and the reaction mixture was allowed to stir at room

- 48 047929 at room temperature for 16 h. The reaction mixture was concentrated in vacuo, then the residue was co-evaporated with DCM (2x20 ml) to give methyl 4-(difluoromethyl)-5-fluoro-III-indole-2-carboxylate I84 (425 mg, 99% yield) as a beige powder.

To a solution of I84 (425 mg, 1.75 mmol) in THF (4 mL) was added a solution of NaOH (210 mg, 5.24 mmol) in water (2 mL). The solution was stirred at room temperature for 18 h. The mixture was diluted with _H2O (60 mL), then washed with DCM (2x30 mL) and EtOAc (30 mL). The aqueous layer was acidified to pH~1 with 1 M aqueous HCl, then extracted with DCM (2x100 mL), DCM with a few drops of MeOH (50 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was dissolved in DCM:MeOH and acidified water; after shaking, the aqueous layer turned pink. The layers were separated and the organic layer was washed with water (3x20 ml), dried over sodium sulfate, filtered and concentrated in vacuo to give the first fraction.

The aqueous layer was extracted with DCM/MeOH 9:1 (3x100 mL), the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give a second fraction. Both fractions were combined and purified by reverse phase flash chromatography (C18_IR_50_F0025, water/ACN 98/2 to 0/100, 50 min) to give 4-(difluoromethyl)-5-fluoro-III-indole-2-carboxylic acid I85 (239 mg, 58% yield) as a beige powder.

3.1.1.12. Synthesis of intermediate compound I90

189

ISO g hydrate hydrazine acetic acid

To a solution of tert-butyl 4-oxopiperidine-1-carboxylate (50 g, 98% purity, 246 mmol) in tetrahydrofuran (500 mL) was added dropwise 1.0 M lithium bis(trimethylsilyl)amide in tetrahydrofuran (250 mL, 250 mmol) at -78 °C. After stirring at -78 °C for 30 min, diethyl oxalate (39.5 g, 270 mmol) in tetrahydrofuran (200 mL) was added dropwise at -78 °C. The mixture was stirred at room temperature overnight. Then the mixture was quenched with water (200 mL) and neutralized with 1 M aqueous hydrochloride solution. The mixture was extracted twice with ethyl acetate (300 mL). _The combined organic phases were washed with brine (500 mL), dried over _Na2SO4 (solid), and filtered. The filtrate was concentrated under reduced pressure to give tert-butyl 3-(2-ethoxy-2-oxoacetyl)-4-oxopiperidine-1-carboxylate (78.4 g, 90% pure by ¹ H NMR, 96% yield) as a yellow oil. 1H NMR (400 MHz, _CDCl3 ) 15.35 (br s, 1H), 4.45 (s, 2H), 4.39 (q, J=7.2 Hz, 2H), 3.66 (t, J=6.0 Hz, 2H), 2.60 (t, J=6.0 Hz, 2H), 1.47 (s, 9H), 1.41 (t, J=6.8 Hz, 3H).

To a solution of tert-butyl 3-(2-ethoxy-2-oxoacetyl)-4-oxopiperidine-1-carboxylate (78.4 g, 90% pure, 236 mmol) in acetic acid (100 mL) was added 85% hydrazine hydrate in water (34 g, 577 mmol) at 0 °C. After stirring at room temperature overnight, the mixture was poured into ice-cold saturated aqueous sodium bicarbonate solution (500 mL). The mixture was extracted twice with ethyl acetate (500 mL). The combined organic phases were washed with brine (1500 mL), dried over Na ₂ SO4 (solid), and filtered. The filtrate was concentrated under reduced pressure to give 5-(tert-butyl)-3-ethyl-1,4,6,7-tetrahydro-5H-niα-zolo[4,3-c]niridine-3,5-dicarboxylate (74.9 g, 90% purity by 1H NMR, 97% yield) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) 13.68 (br s, 0.3H), 13.29 (br s, 0.7H), 4.50 (s, 2H), 4.34-4.19 (m, 2H), 3.61 (t, J=5.6 Hz, 2H), 2.68 (s, 2H), 1.42 (s, 9H), 1.31 (t, J=6.4 Hz, 3H).

To a solution of the intermediate compound 5-(tert-butyl)-3-ethyl-1,4,6,7-tetrahydro-5H-pyrαzolo[4,3c]pyridine-3,5-dicarboxylate (500 mg, 1.524 mmol, purity 90%) in acetonitrile (10 mL) was added

- 49 047929 benzyl (2-bromoethyl)carbamate (415 mg, 1.61 mmol) and cesium carbonate (745 mg, 2.29 mmol). The mixture was stirred at 30°C overnight. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to give intermediate compound I86 (320 mg, yield 42%, purity 95% according to ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.39-7.30 (m, 5H), 5.28 (s, 1H), 5.08 (s, 2H), 4.67-4.64 (m, 2H), 4.53 (s, 2H), 4.32 (q, J=7.2 Hz, 2H), 3.75-3.56 (m, 4H), 2.73-2.71 (m, 2H), 1.49 (s, 9H), 1.37 (t, J=7.2 Hz, 3H).

To a solution of intermediate I86 (320 mg, 95% purity, 0.643 mmol) in ethanol (5 mL) was added 10 wt% palladium on carbon (40 mg). The mixture was heated to 50 °C and stirred overnight over a hydrogen balloon. After cooling to room temperature, the mixture was filtered and the filtrate was concentrated in vacuo to give intermediate I87 (200 mg, 83% yield, 90% purity by LCMS) as a yellow oil. LCMS (ESI): R _t = 1.52 min, mass calcd for C ₁₆ H ₂₆ N ₄ O ₄ 338.2, mass/charge found 339.1 [M+H]+.

To a solution of intermediate I87 (150 mg, 0.399 mmol, 90% purity) in 1,4-dioxane (3 mL) was added 2,3,4,6,7,8-hexahydro-1H-pyrimido[1,2-a]pyrimidine (20 mg, 0.144 mmol). The mixture was stirred at 110 °C for 2 h. The mixture was concentrated to give a residue, which was purified by preparative TLC (dichloromethane:methanol = 10:1) to give intermediate I88 (100 mg, 81% yield, 95% purity by LCMS) as a yellow oil. LCMS (ESI): R _t = 1.39 min, mass, calcd. for C ₁₄ H ₂₀ N ₄ O ₃ 292.2, mass/charge, found value 237.1 [M+H-56]+.

To a solution of intermediate I88 (100 mg, 0.325 mmol, 95% purity) in N,N-dimethylformamide (5 mL) was added sodium hydride (60% in mineral oil, 20 mg, 0.50 mmol). The mixture was stirred at room temperature for 10 min. Then 4-methoxybenzyl chloride (55 mg, 0.35 mmol) was added. The mixture was stirred at room temperature for 1 h. The mixture was poured into water (10 mL) and extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to give intermediate I89 (145 mg, 97% yield, 90% purity by LCMS) as a yellow oil. LCMS (ESI): R _t = 1.62 min, mass calcd. for C ₂₂ H ₂₈ N ₄ O ₄ 412.2, mass/charge, found value 357.1 [M+H-56]+.

To a solution of intermediate I89 (145 mg, 0.316 mmol, 90% purity) in ethyl acetate (2 mL) was added 4 M HCl/EtOAc (3 mL). The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give the hydrochloride salt of intermediate I90 (100 mg, 86% yield, 95% purity by LCMS) as a white solid. LCMS (ESI): R _t = 1.29 min, m/z calcd for C ₁₇ H ₂₀ N ₄ O ₂ 312.2, m/z found 313.1 [M+H]+.

3.1.1.13. Synthesis of intermediates I92*R and I92*S

To a solution of tert-butyl 4-oxo-2-(trifluoromethyl)piperidine-1-carboxylate (4.50 g, 16.84 mmol) in THF (50 mL) was added dropwise LiHMDS (1 M, 20 mL) at -70 °C. The mixture was stirred at -70 °C for 0.5 h. Then diethyl oxalate (3.20 g, 21.89 mmol) was added dropwise at -70 °C. The mixture was stirred at 10 °C for 1 h. The formation of the desired product was mainly observed by LCMS. The mixture was quenched with 1 N HCl (500 mL) and extracted with EtOAc (5 x 300 mL). The combined organic layer was dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo to give the title compound I91 (5.50 g, crude) as a yellow oil, which was used directly in the next step. MS (ESI): mass calcd for C ₁₅ H ₂₀ F ₃ NO ₆ , 367.1; mass/charge found, 368.1 [M+H]+.

To a solution of tert-butyl 5-(2-ethoxy-2-oxoacetyl)-4-oxo-2-(trifluoromethyl)piperidine-1-carboxylate I91 (5.50 g, 14.97 mmol) in EtOH (30 mL) was added NH2NH2-H2O (882 mg, 14.97 mmol, 85% pure). The mixture was stirred at 10 °C for 1 h. By LCMS it was observed that the starting material was consumed, revealing mainly the desired product. The reaction mixture was concentrated in vacuo. The residue was purified by flash chromatography (petroleum ether/EtOAc 80/20 to 60/40) to give the title racemate (4.05 g, 11.15 mmol) as a yellow solid, which was separated by SFC to give two individual enantiomers: (K_*)-5-tert-butyl 3-ethyl-6-(trifluoromethyl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-3,5(4H)-dicarboxylate I92*R (1.40 g, peak 1 on SFC (OD-3S_5_5_40_3ML_T35.M; column: Chiralcel OD-3, 100x4.6 mm ID, particle size 3 μm; mobile phase: 40% EtOH (0.05% DEA) in _CO2 ; flow rate: 3 mL/min; wavelength: 220 nm), retention time=1.002 min) and (*)-5-tert-butyl 3-ethyl-6-(trifluoromethyl)-6,7-dihydro-pyrazolo[4,3-c]pyridine-3,5(4H)-dicarboxylate (1.80 g, peak 2 on SFC (OD-3S_5_5_40_3ML_T35.M; column: Chiralcel OD-3, 100x4.6 mm ID, particle size 3 μm; mobile phase: 40% EtOH (0.05% DEA) in CO2; flow rate: 3 mL/min; wavelength: 220 nm), retention time=1.163 min) as a yellow solid.

3.1.1.14. Synthesis of intermediate compound I99 o

OEt

HCI h ₂ n

OEt

ABOUT

NaOAc, HOAc, NaBH ₃ CN ⁰ meon/osm,0-25 ⁰ C, 24

OEt

193

OEt o

(Sun) ₂ O _ 70°C, 12 h'

Vos

194

OEt t-BuOK, THF 70°C. 1 I.

o o

Sun nh ₂ nh ₂ h ₂ o Jun 70°

I-NH

Vos

N-N

Sun ₂ O, TEA, dcm,25°C, 12 h.

f ₂ hc

DCM, DMAP 0-25°C, 24 h.

6'

F F F F

198

199

The preparation of two batches was carried out in parallel. To a solution of ethyl 3-aminopropanoate hydrochloride compound (332 g, 2.17 mol, 2.00 equiv, HCl) in MeOH (1.50 L) was added NaOAc (177 g, 2.17 mol, 2.00 equiv). The mixture was stirred at 25 °C for 1 h. Then ethyl 4,4-difluoro-3-oxobutanoate (180 g, 1.08 mol, 1.00 equiv) in DCM (2.00 L) was added, followed by HOAc (130 g, 2.17 mol, 123 mL, 2.00 equiv). The mixture was then stirred at 25 °C for 12 h. TLC (petroleum ether/ethyl acetate = 5/1) showed the consumption of the reagent (Rf = 0.30) and the formation of new zones (Rf = 0.50, 0.60). Then a solution of NaBH ₃ CN (102 g, 1.63 mol, 1.50 equiv.) in MeOH (1.00 L) was added portionwise at 0 °C. The mixture was stirred at 20 °C for 12 h. TLC (petroleum ether/ethyl acetate = 5/1) showed the consumption of the starting material (Rf = 0.50, 0.60) and the formation of new zones (Rf = 0.40, 0.10). The two batches were combined for further work-up. The mixture was quenched with a saturated solution of NaHCO ₃ (2.00 L). The organic phases were separated, dried over _Na2SO4 , and concentrated. The crude material was purified by column chromatography on _SiO2 (petroleum ether/ethyl acetate=0/1-10/1). _I93 (420 g, 72.5% yield) was obtained as a light yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.04-5.59 (m, 1H), 4.28-4.03 (m, 4H), 3.28 (tdd, J=3.1, 7.9, 11.2 Hz, 1H), 2.97 (dt, J=3.8, 6.3 Hz, 2H), 2.61 (dd, J=4.8, 16.1 Hz, 1H), 2.52-2.38 (m, 3H), 1.34-1.17 (m, 6H)

A solution of I93 (420 g, 1.57 mol, 1.00 equiv.) and Boc ₂ O (342 g, 1.57 mol, 1.00 equiv.) was stirred at 70 °C for 12 h. Consumption of I93 (Rf = 0.50) and formation of new zones (Rf = 0.70, 0.90) were observed by TLC (petroleum ether/ethyl acetate = 1/0-20/1). The mixture was purified by column chromatography on SiO ₂ (petroleum ether/ethyl acetate = 1/0-20/1). I94 (450 g, yield 77.9%) was obtained as a light yellow oil. 1H NMR (400 MHz, CDCl ₃ ) δ 6.40-5.68 (m, 1H), 4.38-4.06 (m, 5H), 3.74-3.36 (m, 2H), 3.09-2.78 (m, 1H), 2.76-2.48 (m, 3H), 1.47 (br s, 9H), 1.27 (dt, J=4.0, 7.0 Hz, 6H).

Nine batches were prepared in parallel. To a solution of compound I94 (50.0 g, 126 mmol, 1.00 equiv.) in THF (1.50 L) was added t-BuOK (1 M, 177 mL, 1.40 equiv.) in one portion at -70 °C. The mixture was stirred at -70 °C for 1 h. Consumption of compound 3 (Rf = 0.50) and formation of a new zone (Rf = 0.60) were observed by TLC (petroleum ether/ethyl acetate = 5/1). Nine batches were processed in parallel. The mixture was poured into 1 N HCl (500 mL) at 0 °C. The mixture was extracted with MBTE (800 mL x 2). The organic phases were washed with brine (500 mL), dried over Na ₂ SO ₄ , and concentrated. The combined compound I95 (400 g, 100% crude) was combined for the next step without further purification as a light yellow oil. 1H NMR (400 MHz, _CDCl3 ) δ 11.99 (s, 1H), 5.87-5.47 (m, 1H), 4.75-4.42 (m, 1H), 4.22-4.07 (m, 3H), 3.69 (br s, 2H), 2.68-2.45 (m, 2H), 1.45-1.41 (m, 9H), 1.28-1.23 (m, 3H).

The preparation of two batches was carried out in parallel. A mixture of compound I95 (200 g, 572 mmol, 1.00 equiv), TEA (86.9 g, 858 mmol, 119 mL, 1.50 equiv) and NH2NH2-H2O (241 g, 4.10 mol, 234 mL, 85.0% purity, 1.50 equiv) in EtOH (2.00 L) was stirred at 70 °C for 12 h. TLC (petroleum

- 51 047929 ether/ethyl acetate=3/1) consumption of I95 (R=0.80) and formation of a new zone (Rf=0.01) was observed. The two batches were combined for work-up. The reaction mixture was concentrated to give I96 (330 g, 100% crude) as a white solid, which was used directly in the next step.

I96 (144 g, 497 mmol, 1.00 equiv), TEA (75.6 g, 746 mmol, 103 mL, 1.50 equiv) and _Boc2O (130 g, 597 mmol, 137 mL, 1.20 equiv) in DCM (2.00 L) were stirred at 25 °C for 12 h. Consumption of I96 ( _Rf 0.01) and formation of a new zone (Rf = 0.40) was observed by TLC (petroleum ether/ethyl acetate = 1/1). Complete reaction was observed by HPLC (EW15208-75-P1A). The mixture was concentrated to approximately 150 mL and a white solid precipitated. MBTE (150 mL) was added. The mixture was stirred at 20 °C for 1 h. The white solid was filtered to afford compound I97 (120 g, 60.6% yield) as a white solid. By LCMS (EW15208-75-P1G3, RT=15.253 min) it was observed that the regioisomer was completely separated. 1H NMR (400 MHz, _CDCl3 ) δ 11.17 (br s, 1H), 6.46-5.89 (m, 1H), 5.03-4.62 (m, 1H), 4.50 (br d, J=15.9 Hz, 1H), 3.95-3.62 (m, 1H), 3.23-2.99 (m, 2H), 1.53 (s, 9H), 1.44 (s, 9H)

To a mixture of compound 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonyl fluoride (139 g, 462 mmol, 81.9 mL, 1.50 equiv), compound I97 (120 g, 308 mmol, 1.00 equiv), and DMAP (1.88 g, 15.41 mmol, 0.05 equiv) in DCM (1.00 L) was added DIPEA (119 g, 924 mmol, 161 mL, 3.00 equiv) at 0°C. The mixture was stirred at 25°C for 24 h. TLC (petroleum ether/ethyl acetate = 3/1) showed almost complete consumption of compound I97 (Rf = 0.20) and formation of new zones (Rf = 0.60, 0.10). The mixture was washed with water (500 ml) and concentrated. The residue was purified by column chromatography on SiO ₂ (petroleum ether/ethyl acetate = 50/1-10/1). Compound I98 (105 g, yield 50.2%) was obtained as a colorless oil.

An autoclave was charged with compound I98 (105 g, 156 mmol, 1.00 equiv), DPPP (6.45 g, 15.6 mmol, 0.10 equiv), Pd(OAc) ₂ (3.51 g, 15.64 mmol, 0.10 equiv), and TEA (31.6 g, 312 mmol, 43.5 mL, 2.00 equiv) in EtOH (1.20 L) and DMF (1.20 L). The mixture was then purged three times with nitrogen and three times with carbon monoxide. The mixture was stirred at 90°C under CO atmosphere (0.5 MPa) for 48 h. By TLC (petroleum ether/ethyl acetate=1/1) the consumption of I98 (Rf=0.80) and the formation of new zones (Rf=0.30, 0.0) were observed. The filtrate was concentrated. The residue was purified by chromatography on _SiO2 (petroleum ether/ethyl acetate=5/1-1/1) to give I99 (26.0 g, yield 34.5%) as a yellow solid. 1H NMR (400 MHz, CDCl ₃ ) δ 12.50-11.55 (m, 1H), 5.94-5.47 (m, 1H), 5.28-4.72 (m, 2H), 4.51-4.15 (m, 3H), 3.222.99 (m, 2H), 1.52 (s, 9H), 1 .45-1.36 (m, 3H).

3.1.1.15. Synthesis of intermediate compound I101

1100

1101

4-(Difluoromethoxy)acetophenone (5 g, 26.86 mmol), and 3-5] and 2-aminoethan-1-ol (4.8 mL, 80.58 mmol) were dissolved in dry MeOH (62.5 mL) under nitrogen atmosphere. Ti(iPrO) ₄ (10.2 mL, 34.92 mmol) was added and the mixture was stirred at 65 °C for 16 h. The mixture was cooled to rt and further cooled with an ice bath. Sodium borohydride (1.02 g, 26.86 mmol) was added portionwise and the mixture was stirred at rt for 1 h. The reaction mixture was quenched by addition of water and stirring was continued at rt for 20 min. The mixture was acidified with 1 N HCl. The mixture was filtered through a pad of celite and washed with water and EtOAc. The filtrate was concentrated in vacuo. The residue was dissolved in water and neutralized with _NaHCO3 . The mixture was extracted with Me-THF, dried over _MgSO4 , filtered and concentrated in vacuo to give 2-((1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethan-1-ol I100 (6.1 g, 98.214% yield) as an oil.

To a solution of 2-((1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethan-1-ol (6.1 g, 26.38 mmol) and _NaHCO3 (8.8 g, 105.52 mmol) in THF (60 mL) and water (75 mL) was added BOC anhydride (15.8 mL, 1 M in THF, 31.65 mmol). The mixture was stirred at rt for 16 h. THF was removed. The mixture was extracted twice with Me-THF. The combined organic layers were dried over _MgSO4 , filtered and concentrated. The residue was purified by column chromatography (heptane/EtOAc from 100/5 to 40/60) to give I101 (6 g, 68% yield).

- 52 047929

3.1.1.16. Synthesis of intermediate compound I103

To a solution of 2-pyridinecarboxaldehyde (9 mL, 94.611 mmol) in EtOH (150 mL) was added ethanolamine (5.7 mL, 94.61 mmol) and the reaction mixture was stirred at rt for 16 h. Sodium borohydride (3.6 g, 96.50 mmol) was then added partly at 0 °C and the reaction mixture was stirred at rt for 1.5 h. The reaction mixture was quenched with saturated _NaHCO3 solution (100 mL) and stirred at rt for 30 min. The resulting white solid was filtered off and the filtrate was evaporated to give crude 2-((pyridin-2-ylmethyl)amino)ethan-1-ol I102 (18.31 g) as an orange oil, which was used as such in the next step.

Crude 2-((pyridin-2-ylmethyl)amino)ethan-1-ol I102 (1.44 g, 9.46 mmol) was dissolved in THF (20 mL) and water (26 mL). Then _NaHCO3 (3.18 g, 37.84 mmol) was added followed by di-tert-butyl dicarbonate (5.7 mL, 2 M, 11.35 mmol). The reaction mixture was stirred at rt for 2 h. The formed solids/salts were filtered off and THF was separated. The aqueous layer was extracted once more with EtOAc and the organic layers were combined and evaporated under reduced pressure. The crude material was purified by column chromatography (heptane/EtOAc 100/0 to 0/100) to give intermediate I103 (21.7 g, 85% yield) as a yellow oil.

3.1.1.17. Synthesis of intermediate compound I106

To a solution of I103 (21.2 g, 28 mmol) in dry THF (510 mL) were added tert-butyl (1-(4-(difluoromethoxy)phenyl)ethyl)(2-hydroxyethyl)carbamate (26 g, 28 mmol) and triphenylphosphine (46.4 g, 168 mmol). The reaction mixture was sealed at 10 atm. Then DIAD (32.7 mL, 168 mmol) was added and the mixture was stirred at rt for 16 h. The solvents were evaporated and the crude material was redissolved in DIPE and _saturated NaHCO3 was added. The mixture was stirred for 15 min. The resulting solid was filtered off. The organic layer was separated and evaporated under reduced pressure. The brown oil was purified by column chromatography (heptane/(EtOH:EtOAc 10/30) 100/0 to 0/100). The residue was triturated in DIPE. The suspension was stirred at rt overnight. The solid was filtered off and the filtrate was evaporated to give an orange oil, 5-(tert-butyl)-3-ethyl-(4-c)-2-(2-((tert-butoxycarbonyl)(pyridin-2-ylmethyl)amino)ethyl)-6-methyl-2,4,6,7-tetrahydro-5H-pyrα-zolo[4,3-c]pyridine-3,5-dicarboxylate I104 (75 g, 71% yield), which was used as such in the next step.

A flask was charged with 5-(tert-butyl)-3-ethyl-(3)-2-(2-((tert-butoxycarbonyl)(pyridin-2-ylmethyl)amino)ethyl)-6-methyl-2,4,6,7-tetrahydro-5H-pyrαzolo[4,3-c]pyridine-3,5-dicarboxylate I104 (20 g, 15.2 mmol) in dry 1,4-dioxane (94 mL). HCl (40 mL, 4 M in dioxane, 158 mmol) was added and the mixture was stirred at rt for 3 days. The mixture was concentrated in vacuo. The product was coevaporated with diethyl ether and the resulting yellowish foam was triturated with diethyl ether. The resulting yellowish solid was filtered off and dried in vacuo at 45°C overnight to give ethyl (R)-6-methyl-2-(2-((pyridin-2-ylmethyl)amino)ethyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate HCl I105 (10.4 g, 52% purity, 82% yield). The filtrate was evaporated and the residue was triturated again in ether. The precipitate was filtered off to give a second fraction I105 (beige solid) (1.5 g, 52% purity, 11% yield), which was dried in vacuo overnight at 45°C. The crude fractions were used as such in the next step.

A pressure vial was charged with I105 (10.4 g, 13 mmol) in toluene (24 mL). 1,5,7-Triazabicyclo[4.4.0]ec-5-ene (TBD) (7.22 g, 51.8 mmol) was added and the vial was capped at 100 atm. The mixture was stirred at 100 °C for 2 h. The mixture was cooled and concentrated in vacuo. The product was purified

- 53 047929 by column chromatography (DCM/[MeOH/7 M NH3] 100/0 to 90/10) to give (R)-3methyl-9-(pyridin-2-ylmethyl)-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one I106 (5 g, quantitative yield) as a yellow oil, which solidified after standing at rt.

3.1.1.18. Synthesis of intermediate compound I109

_MeSO2CI

_Et3N , DCM

THF

Vos ₂ O, Et ₃ N, DCM

1107

1108

MsCl (99.8 g, 871 mmol) was added dropwise to a solution of 3-butyn-2-ol (50.4 g, 719 mmol), triethylamine (108 g, 1075 mmol), DCM (504 mL) over 30 min in an ice bath. The reaction mixture was stirred for 2 h at room temperature, then washed with water (500 mL). The organic layer was dried over sodium sulfate, filtered and concentrated to give 2-(but-3-yn-2ylamino)ethan-1-ol I107 as a pale oil, which was used as such in the next step.

I107 (112 g, 753 mmol), 2-aminoethan-1-ol (115 g, 1882 mmol) in THF (750 mL) was heated under reflux over the weekend. The reaction mixture was concentrated in vacuo, and sat. aq. _NaHCO3 (300 mL) was added, and the mixture was extracted with _Et2O (3x300 mL). _The combined organic phases were dried over _Na2SO4 and evaporated to dryness. The residue was purified by column chromatography (heptane/EtOAc from 90/10 to 0/100) to give 2-(but-3-yn-2-ylamino)ethan-1-ol I108 (37 g, 43% yield) as a clear oil.

_Et3N (59 mL, 426 mmol) was added to a solution of 2-(but-3-yn-2-ylamino)ethan-1-ol I108 (37 g, 328 mmol) and di-tert-butyl dicarbonate (86 g, 393 mmol) in DCM (500 mL) and stirred overnight. The reaction mixture was concentrated. The residue was purified by column chromatography (DCM/MeOH 100/0 to 80/20) to give tert-butyl but-3-yn-2-yl(2-hydroxyethyl)carbamate I109 (53 g, 75%).

3.1.1.18. Synthesis of intermediate compound 1111 ^NH НС1, dioxane _Н| ^

N

DECP, _Et3N

To a stirred solution of 5-(tert-butyl)-3-ethyl-(R)-6-methyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (20 g, 64.7 mmol) in DCM (200 mL) was added HCl (81 mL, 4 M in dioxane, 323 mmol). The reaction mixture was then stirred at rt overnight. After one night, the reaction mixture was evaporated and the residue was co-evaporated twice with diethyl ether. The residue was triturated in diethyl ether (200 ml) and the precipitate was filtered and washed with diethyl ether (2 x 50 ml) and dried in a vacuum oven for 3 h to give ethyl (((4,3-c)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I110 (18 g, quantitative yield) as a yellow powder, which was used as such in the next step.

A vial was charged with (R)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I110 (1 g, 3.95 mmol) and _Et3N (1.6 mL, 11.8 mmol) in DCM (20 mL). 4-Chloro-3-cyanobenzoic acid (788 mg, 4.34 mmol) was added and the vial was capped. Diethyl cyanophosphonate (0.85 mL, 5.13 mmol) was added dropwise and the mixture was stirred at rt for 16 h. 10 mL of semi-sat. aq. _NaHCO3 was added. After vigorous stirring for 5 min, the layers were separated. The organic layer was dried over _MgSO4 , filtered and concentrated in vacuo. The residue was purified by column chromatography (heptane/EtOAc 100/0 to 0/100) to give ethyl ((3-c)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I111 (836 mg, 57% yield) as a foam.

3.1.2. Synthesis of compounds.

3.1.2.1. Synthesis of compound 1

- 54 047929

Intermediate connection I10.

Ethyl 2-(2-aminoethyl)-5-(3,4-dichlorobenzoyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3carboxylate 2TFA

L 1 N ' *2TFA ''Π'' q CO ₂ Et

110

TFA (4.49 mL, 58.7 mmol) was added to a solution of intermediate I5 (3.00 g, 5.87 mmol) in DCM (100 mL). The reaction mixture was stirred at room temperature overnight and concentrated under reduced pressure to give intermediate I10, which was used as such in the next step.

Connection 1.

2-(3,4-Dichlorobenzoyl)-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

Intermediate I10 was dissolved in EtOAc (150 mL). The solution was washed with _NaHCO3 (sat., aq., 150 mL), dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The residue was diluted in THF (5 mL) and MeCN (1 mL) and TBD (245 mg, 1.76 mmol) was added. The reaction mixture was heated at 100 °C for 105 min in a Biotage microwave. The reaction mixture was cooled to room temperature, whereupon a precipitate formed overnight. The white crystals were filtered, washed with MeCN and dried at 50 °C under vacuum to afford compound 1 (872 mg, 41% over 2 steps).

1H NMR (400 MHz, DMSO-d ₆ , 101°C) δ ppm 7.78 (br s, 1H), 7.67 (d, J=8.1 Hz, 1H), 7.65 (d, J=2.0 Hz, 1H), 7.40 (dd, J=8.3, 1.9 Hz, 1H), 4.66 (s, 2H), 4.17-4.23 (m, 2H), 3.64-3.82 (m, 2H), 3.55-3.62 (m, 2H), 2.74 (t, J=5.9 Hz, 2H); LCMS (method B): Rt=1.53 min, m/z calcd. for C ₁₆ H ₁₄ Cl ₂ N ₄ O ₂ 364.0, the found value is 365 [M+H]+.

3.1.2.2. Synthesis of compound 2

Intermediate connection I11.

5-tert-Butyl-3-ethyl-2-{2-[(propan-2-yl)amino]ethyl}-2,4,6,7-tetrahydro-5H-pyrazolo[4,3c]pyridine-3,5-dicarboxylate

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A mixture of intermediate I6 (0.70 g, 1.74 mmol), isopropylamine (1.03 g, 17.4 mmol), _K2CO3 (481 mg, 3.48 mmol) and NaI (26.1 mg, 0.17 mmol) in MeCN (115 _mL ) was stirred at reflux for 6 h. The reaction mixture was filtered through decalite and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica, heptane/(EtOAc/(EtOH/7 M aq. _NH3 in MeOH), 75/25/2), gradient from 30:70 to 0:100) to give intermediate I11 (105 mg, 16%) as a brown oil.

Intermediate connection I12.

tert-Butyl 10-oxo-9-(propan-2-yl)-3,4,7,8,9,10-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazine2(III)-carboxylate

A mixture of intermediate I11 (105 mg, 0.276 mmol) and TBD (11.5 mg, 82.8 μmol) in THF (2 mL) was stirred at 100 °C overnight. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (silica, heptane/EtOAc, gradient from 80:20 to 0:100) to give intermediate I12 (82 mg, 89%).

Connection 2.

2-(3,4-Dichlorobenzoyl)-9-(propan-2-yl)-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

TFA (0.19 mL, 2.45 mmol) was added to a solution of intermediate I12 (82 mg, 0.24 mmol) in DCM (30 mL) and the reaction mixture was stirred overnight. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM (30 mL). DIPEA (0.169 mL, 0.98 mmol) was added followed by 3,4-dichlorobenzoyl chloride (51.4 mg, 0.245 mmol). The reaction mixture was stirred at room temperature for 1 h and the mixture was subjected to flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 70:30 to 0:100) to give compound 2 (82 mg, 82%).

¹ H NMR (400 MHz, DMSC-d6, 101°C) δ ppm 7.67 (d, J=8.1 Hz, 1H), 7.65 (d, J=2.0 Hz, 1H), 7.40 (dd, J=8.4, 2.0 Hz, 1H), 4.61-4.72 (m, 3H), 4.21-4.27 (m, 2H), 3.67-3.77 (m, 2H), 3.61-3.66 (m, 2H), 2.73 (t, J=5.9 Hz, 2H), 1.15 (d, J=6.8 Hz, 6H); LCMS (method D): R _t =1.85 min, m/z calc. for C ₁₉ H ₂₀ Cl ₂ N ₄ O ₂ 406.1, the found value is 407.2 [M+H]+; mp 190°C.

3.1.2.3. Synthesis of compound 3.

9-Benzyl-2-(3,4-dichlorobenzoyl)-1,2,3,4,8,9-hexahydronipirogolo[4',3':3,4]nipirazolo[1,5-a]nipirazin10(7H)-one

To a solution of compound 1 (125 mg, 0.34 mmol) in anhydrous DMF (4.1 mL) was added NaH (60% dispersion in mineral oil, 20.5 mg, 0.51 mmol) at room temperature. The reaction mixture was stirred for 15 min and benzyl bromide (44.8 μL, 0.38 mmol) was added. The reaction mixture was stirred at room temperature for an additional 2 h. The reaction mixture was quenched with water and diluted with MeOH. The solvents were evaporated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 90:10) to give compound 3 (145 mg, 93%) as a glassy white solid.

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1H NMR (400 MHz, DMSO-d ₆ , 81°C) δ ppm 7.67-7.71 (m, 2H), 7.43 (dd, J=8.1, 2.0 Hz, 1H), 7.24-7.37 (m, 5H), 4.71 (br s, 2H), 4.64 (s, 2H), 4.25-4.29 (m, 2H), 3.63-3.82 (m, 4H), 2.74 (t, J=5.8 Hz, 2H) LCMS (method C): R _t = 1.05 min, m/z calcd for C2 ₃ H ₂₀ Cl ₂ N ₄ O2 454.0, found 455.3 [M+H]+.

3.1.2.4. Synthesis of compound 4.

2-(3,4-Dichlorobenzoyl)-9-[(4-methoxyphenyl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1 D^pyrazin-1 0(7H)-one

Compound 4 was prepared according to the procedure described for the synthesis of compound 3.

The crude mixture was purified by flash column chromatography (silica, hexane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100) to afford compound 4 (153 mg, 95%).

1H NMR (400 MHz, DMSO-d6, 101°C) δ ppm 7.68 (d, J=8.4 Hz, 1H), 7.66 (d, J=2.0 Hz, 1H), 7, 41 (dd, J=8.1, 2.0 Hz, 1H), 7.20-7.27 (m, 2H), 6.85-6.93 (m, 2H), 4.70 (s, 2H), 4.57 (s, 2H), 4.21-4.27 (m, 2H), 3.74 (s, 3H), 3.69-3.73 (m, 2H), 3.61 -3.67 (m, 2H), 2.74 (t, J=5.8 Hz, 2H); LCMS (method B): Rt=1.98 min, m/z _calcd for _{C24H22Cl2N4O3 484.0} , found value 485.1 [M+H] ⁺ .

3.1.2.5. Synthesis of compound 5.

2-(3,4-Dichlorobenzoyl)-9-[1-(pyridin-4-yl)ethyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1 D^pyrazin-1 0(7H)-one

_NaBH3CN

MeOH CI

k.t., 5 days * CI

2) TBD

THF

100°С, o/WE

110

Intermediate I10 (1.06 g, 1.66 mmol) and 4-acetylpyridine (605 mg, 4.99 mmol) were dissolved in MeOH (20 mL). The mixture was stirred at room temperature and sodium cyanoborohydride (1.10 g, 16.6 mmol) was added portionwise twice daily for 5 days. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc and washed with _Na2CO3 ( _aq ). The organic layer was dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100). The residue was dissolved in THF (2 mL) in a microwave tube. TBD (69.5 mg, 0.50 mmol) was added and the reaction mixture was heated at 100 °C over the weekend. The mixture was concentrated under reduced pressure and the crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100). The residue was dried overnight under vacuum at 50 °C to give compound 5 (18.2 mg, 2.3%) as a white solid.

1H NMR (400 MHz, DMSO-d ₆ , 101°C) δ ppm 8.51-8.54 (m, 2H), 7.68 (d, J=8.4 Hz, 1H), 7.66 ( d, J=2.0 Hz, 1H), 7.41 (dd, J=8.1, 2.0 Hz, 1H), 7.30-7.33 (m, 2H), 5.73 (q , J=7.2 Hz, 1H), 4.70 (s, 2H), 4.18-4.32 (m, 2H), 3.65-3.79 (m, 3H), 3.36- 3.44 (m, 1H), 2.74 (t, J=5.8 Hz, 2H), 1.56 (d, J=7.0 Hz, 3H); LCMS (method D): R _t = 1.73 min, mass/charge calc. for C2 ₃ H ₂₁ Cl ₂ N ₅ O2 469.1, found value 470.2 [M+H] ⁺ .

3.1.2.6. Synthesis of compound 6.

2-(3,4-Dichlorobenzoyl)-9-[(2-methoxypyridin-4-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[^^pyrazin- 10(7H)-one

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Compound 6 was prepared according to the procedure described for the synthesis of compound 3.

The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc, gradient from 100:0 to 0:100). A second purification was achieved by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 μm, 30x150 mm, mobile phase: _NH4HCO3 (0.25% in water)/MeCN) to afford compound 6 (59 mg, 37%) as a white solid.

¹ HNMR (600 MHz, DMSO-d6, 83°C) δ ppm 7.66-7.70 (m, 2H), 7.64 (t, J=7.7 Hz, 1H), 7.42 (dd , J=8.3, 1.8 Hz, 1H), 6.90 (br d, J=7.1 Hz, 1H), 6.67 (d, J=8.3 Hz, 1H), 4, 59-4.74 (m, 4H), 4.33 (t, J=6.0 Hz, 2H), 3.87 (br t, J=6.0 Hz, 2H), 3.80 (s, 3H), 3.57-3.83 (m, 2H), 2.75 (br t, J=5.5 Hz, 2H); LCMS (method C): R _t = 1.01 min, mass/charge calcd. for C ₂₃ H ₂₁ Cl ₂ N ₅ O ₃ 485.1, found value 486.2 [M+H]+.

3.1.2.7. Synthesis of compound 7.

2-(3,4-Dichlorobenzoyl)-9-[(1,3-thiazol-2-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5^]pyrazin-1 0(7H)-one

1) NaH

Compound 7 was prepared according to the procedure described for the synthesis of compound 3.

The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc, gradient from 100:0 to 0:100). A second purification was achieved by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 μm, 30x150 mm, mobile phase: NH4HCO3 (0.25% in water)/MeCN) to afford compound 7 (99 mg, 65% yield) as a white solid.

¹ H NMR (400 MHz, DMSO-d ₆ , 81°C) δ ppm 7.73 (d, J=3.3 Hz, 1H), 7.66-7.71 (m, 2H), 7.63 (d, J=3.3 Hz, 1H), 7.42 (dd, J=8.1, 2.0 Hz, 1H), 4.95 (s, 2H), 4.69 (br s, 2H), 4.28-4.36 (m, 2H), 3.84-3.89 (m, 2H), 3.63-3.80 (m, 2H), 2.75 (t, J=5.8 Hz, 2H); LCMS (method C): R _t = 0.88 min, m/z calc. for C ₂₀ H ₁₇ Cl ₂ N ₅ O ₂ S 461.0, the found value is 462.2 [M+H]+.

3.1.2.8. Synthesis of compound 8.

2-(3,4-Dichlorobenzoyl)-9-[(1H-indol-3-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo [1.5^]pyrazin-1 0(7H)-one

1)NaH

To a mixture of compound 1 (208 mg, 0.57 mmol) in DMF (5 mL) was added NaH (60% dispersion in mineral oil, 34.2 mg, 0.85 mmol) and the reaction mixture was stirred for 15 min at room temperature. tert-Butyl 3-(bromomethyl)-1/3-indole-1-carboxylate (194 mg, 0.63 mmol) was added and the reaction mixture was stirred for an additional 2 h. The reaction mixture was quenched with MeOH (1 mL) and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100). The product was dissolved in DCM (10 mL) and TFA (0.44 mL, 5.70 mmol) was added. The reaction mixture was stirred at room temperature overnight. The mixture was washed with _NaHCO3 (sat., aq., 250 mL), dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100) to give compound 8 (41 mg, 15%) as a light yellow gum.

1H NMR (400 MHz, DMSO-d ₆ , 101°C) δ ppm 10.75 (s, 1H), 7.69 (d, J=8.1 Hz, 1H), 7.67 (d, J= 2.0 Hz, 1H), 7.56 (d, J=7.9 Hz, 1H), 7.42 (dd, J=8.1, 2.0 Hz, 1H), 7.36 (d, J=8.1 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.047.10 (m, 1H), 6.93-6.98 (m, 1H), 4, 79 (s, 2H), 4.74 (s, 2H), 4.13-4.18 (m, 2H), 3.66-3.80 (m, 2H), 3.59-3.65 (m, 2H), 2.73 (t, J=5.8 Hz, 2H); LCMS (method B): R _t =1.91 min, mass/charge calc. for C ₂₅ H ₂₁ Cl ₂ N ₅ O ₂ 493.1, found value 494.1 [M+H]+.

3.1.2.9. Synthesis of compound 9.

2-(3,4-Dichlorobenzoyl)-9-[(III-indol-4-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo [1.5^]pyrazin-1 0(7H)-one

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Compound 9 was prepared according to the procedure described for the synthesis of compound 3.

The crude mixture was purified by flash column chromatography (silica, DMC/MeOH, gradient 100:0 to 90:10). A second purification was accomplished by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 μm, 30x150 mm, mobile phase: _NH4HCO3 (0.25% in water)/MeCN). The residue was triturated in DIPE. The solids were collected by filtration to afford compound ₉ (109 mg, 64%) as a grayish solid.

^1H NMR (400 MHz, DMSO-d6, 81°C) δ ppm 10.96 (s, 1H), 7.71 (d, J=6.4 Hz, 1H), 7.69 (s, 1H) , 7.42-7.46 (m, 1H), 7.35 (d, J=8.1 Hz, 1H), 7.28 (t, J=2.8 Hz, 1H), 7.05 ( t, J=7.6 Hz, 1H), 6.95 (d, J=7.0 Hz, 1H), 6.48 (br s, 1H), 4.90 (s, 2H), 4.66 -4.83 (m, 2H), 4.12-4.21 (m, 2H), 3.64-3.86 (m, 2H), 3.56-3.62 (m, 2H), 2.74 (t, J=5.7 Hz, 2H); LCMS (method C): R _t =0.97 min, m/z calcd for C ₂₅ H ₂₁ Cl ₂ N ₅ O ₂ 493.1, found value 494.3 [M+H]+.

3.1.2.10. Synthesis of compound 10.

2-(3,4-Dichlorobenzoyl)-9-{[4-(difluoromethoxy)phenyl]methyl}-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-1 0(7H)-one

Compound 10 was prepared according to the procedure described for the synthesis of compound 3.

The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc, gradient 100:0 to 0:100) to afford compound 10 (147 mg, 85%) as a white foam.

1H NMR (400 MHz, DMSO-d ₆ , 81°C) δ ppm 7.69 (d, J=8.5 Hz, 1H), 7.67 (d, J=2.0 Hz, 1H), 7.42 (dd, J=8.3, 1.8 Hz, 1H), 7.34-7.39 (m, 2H), 7.11-7.16 (m, 2H), 7.09 (t, J=74.2 Hz, 1H), 4.66-4.78 (m, 2H), 4.63 (s, 2H), 4.24-4.30 (m, 2H), 3.66-3.84 (m, 4H), 2.74 (t, J=5.7 Hz, 2H).

3.1.2.11. Synthesis of compound 11

1)NaH

Cl

Cl

Intermediate compound I13.

Ethyl (6R)-2-(2-aminoethyl)-5-(3,4-dichlorobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate hydrochloride

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

HCl (4N in 1,4-dioxane, 24.8 ml, 99.0 mmol) was added dropwise to a solution of intermediate I8 (3.47 g, 6.60 mmol) in DCM (60 ml) at room temperature and the reaction mixture was stirred for 2 h. The mixture was concentrated to dryness and co-evaporated with DCM (2 x 40 ml) to give intermediate I13 as a white solid, which was used as such in the next step.

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Intermediate compound I14 (C251).

(3R)-2-(3,4^uxnop6eH3Oun)-3-MeTua-1,2,3,4,8,9-reKcarugponupugo[4',3':3,4]nupa3oao[1,5-a] nupa3UH10(7H)-on

114

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I13 in DCE (110 mL) was added AlMe ₃ (2 M in toluene, 9.91 mL, 19.8 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 1 h and at 80 °C for another h. The reaction mixture was cooled to room temperature and diluted with DCM (50 mL). Na ₂ SO ₄ -10H ₂ O (~10 g) was added portionwise. The mixture was stirred for 5 min and anhydrous Na ₂ SO ₄ was added. The suspension was filtered and washed with DCM (50 mL). The filtrate was washed with NaHCO ₃ (sat., aq., 150 mL), HCl (1 M, aq., 150 mL), and brine (150 mL). The organic layer was dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5) to give intermediate I14 ( _2.20 g, 85% over 2 steps) as a white solid.

Connection 11.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(4-methoxyphenyl)methyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-1 0(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I14 (350 mg, 0.92 mmol) in DMF (15 mL) at 0 °C was added NaH (60% in mineral oil, 55.4 mg, 1.38 mmol). The reaction mixture was stirred at room temperature for 30 min, then p-methoxybenzyl bromide (204 mg, 1.02 mmol) was added. The reaction mixture was stirred at room temperature for 2 h and diluted with EtOAc (100 mL), washed with H ₂ O (3 x 100 mL), brine, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 97:3). The residue was solubilized in EtOAc (100 mL) and the solution was washed successively with _H2O (3x100 mL) and brine (100 mL), dried ( _{Na2SO4 )} , filtered and concentrated under reduced pressure to give compound 11 (391 mg, 84%).

^1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.72 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.7 Hz, 1H), 7 .43 (dd, J=8.2, 1.7 Hz, 1H), 7.27 (d, J=8.5 Hz, 2H), 6.82 (d, J=8.5 Hz, 2H) , 5.10 (br.s, 1H), 4.82-4.54 (m, 3H), 4.39-4.22 (m, 3H), 3.76 (s, 3H), 3.68 (t, J=5.9 Hz, 2H), 2.95 (dd, J=16.2, 5.6 Hz, 1H), 2.58 (d, J=16.2 Hz, 1H), 1.18 (d, J=6.6 Hz, 3H); LCMS (method A): Rt=10.5 min, m/z calcd for C ₂₅ H ₂₄ Cl ₂ N ₄ O ₃ 498, mass/charge, found value 499 [M+H]+.

3.1.2.12. Synthesis of compound 12.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(4-fluorophenyl)methyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-1 0(7H)-one

k.t., 2 hours

Compound 12 was prepared according to the procedure described for the synthesis of compound 11.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 97:3). The residue was solubilized in EtOAc (100 mL) and washed successively with _H2O (3x100 mL) and brine (100 mL), dried ( _{Na2SO4 )} , filtered and concentrated under reduced pressure to give compound 12 (360 mg, 86%).

1H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 7.72 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.7 Hz, 1H), 7 .43 (dd, J=8.2, 1.7 Hz, 1H), 7.41-7.36 (m, 2H), 7.19-7.12 (m, 2H), 5.19-4 .98 (m, 1H), 4.80-4.59 (m, 3H), 4.36-4.24 (m,

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3H), 3.72 (t, J=6.1 Hz, 2H), 2.95 (dd, J=15.8, 5.9 Hz, 1H), 2.58 (d, J=15.8 Hz, 1H), 1.18 (d, J=6.7 Hz, 3H); LCMS (method A): Rt=10.7 min, m/z calcd for C ₂₄ H ₂₁ Cl ₂ FN ₄ O ₂ 486, m/z found 487 [M+H]+.

3.1.2.13. Synthesis of compound 13.

(3R)-2-(3,4-Dichlorobenzoyl)-9-{[4-(difluoromethoxy)phenyl]methyl}-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-1 0(7H)-one

Compound 13 was prepared according to the procedure described for the synthesis of compound 11.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 97:3). The residue was solubilized in EtOAc (100 mL) and washed successively with _H2O (3x100 mL) and brine (100 mL), dried ( _{Na2SO4 )} , filtered and concentrated under reduced pressure to give compound 13 (375 mg, 80%).

1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.72 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.7 Hz, 1H), 7, 43 (dd, J=8.2, 1.7 Hz, 1H), 7.40 (d, J=8.3 Hz, 2H), 7.16 (d, J=8.3 Hz, 2H), 7.14 (t, J=74.0 Hz, 1H), 5.08 (br.s, 1H), 4.804.52 (m, 3H), 4.38-4.23 (m, 3H), 3 .73 (t, J=6.2 Hz, 2H), 2.96 (dd, J=15.6, 5.4 Hz, 1H), 2.58 (d, J=15.6 Hz, 1H), 1.18 (d, J=6.4 Hz, 3H); LCMS (method A): R _t =10.8 min, m/z calc. . for C ₂₅ H ₂₂ Cl ₂ F ₂ N ₄ O ₃ 534, mass/charge, the found value is 535 [M+H]+.

3.1.2.14. Synthesis of compound 14.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-{[4-(trifluoromethoxy)phenyl]methyl}-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-α]pyrazin-10(7H)-one

Cl

Cl

1)NaH DMF

Ν~\ΟΤ (GS DO K.T., 30

Compound 14 was prepared according to the procedure described for the synthesis of compound 11.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 97:3). The residue was solubilized in EtOAc (100 mL) and the mixture was washed successively with _H2O (3x100 mL) and brine (100 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure to give compound 14 ( ₃₈₅ mg, 80%).

1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.72 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.7 Hz, 1H), 7, 48 (d, J=8.4 Hz, 2H), 7.43 (dd, J=8.2, 1.7 Hz, 1H), 7.32 (d, J=8.4 Hz, 2H), 5.09 (br.s, 1H), 4.87-4.52 (m, 3H), 4.38-4.23 (m, 3H), 3.75 (t, J=6.2 Hz, 2H), 2.96 (dd, J=16.0, 6.0 Hz, 1H), 2.59 (d, J=16.0 Hz, 1H), 1.18 (d, J=6.4 Hz, 3H); LCMS (method A): R _t =11.4 min, m/z calc. for C ₂₅ H ₂₁ Cl ₂ F ₃ N ₄ O ₃ 552, mass/charge, found value 553 [M+H]+.

3.1.2.15. Synthesis of compound 15.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(pyridin-2-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

NaH (60% in mineral oil, 66.4 mg, 1.66 mmol) was added to a solution of intermediate I14 (130 mg, 0.33 mmol) in DMF (4 mL) and the reaction mixture was stirred at room temperature for 1 h. 2-Picolyl chloride (109 mg, 0.66 mmol) was added and the reaction mixture was stirred at room temperature overnight. The mixture was combined with another fraction (0.05 mmol) and diluted with H ₂ O (40 mL). The mixture was extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with NaHCO ₃ (sat., aq., 50 mL) and brine (3 x 50 mL), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture

- 61 047929 was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 90:10). A second purification was achieved by reversed-phase flash column chromatography (C-18, water/MeCN, gradient from 75:25 to 25:75) to give compound 15 (90 mg, 50%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 8.52 (ddd, J=4.8, 1.6, 0.8 Hz, 1H), 7.76 (td, J=7.6, 2.0 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.69 (d, J=2.0 Hz, 1H), 7.42 (dd, J=8.0, 2.0 Hz, 1H), 7.36 (d, J=7.6 Hz, 1H), 7.28 (ddd, J=8.0, 5.2, 0.8 Hz, 1H), 5.05 (br.s, 1H), 4.80-4.72 (m, 2H), 4.63 (br.s, 1H), 4.39-4.26 (m, 3H), 3.90-3.80 (m, 2H), 2.95 (dd, J=16.0, 6.0 Hz, 1H), 2.58 (d, J=16.0 Hz, 1H), 1.17 (d, J=6.4 Hz, 3H); LCMS (method A): R _t = 8.6 min, m/z calcd for C ₂₃ H ₂₁ Cl ₂ N ₅ O2 469, m/z found 470 [M+H]+.

3.1.2.16. Synthesis of compound 16.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(pyridin-3-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrαzolo[1,5-a]pyrαzin-10(7H)-one

Compound 16 was prepared according to the procedure described for the synthesis of compound 15.

The crude mixture was purified by flash column chromatography (silica,

DCM/MeOH, gradient from 100:0 to 90:10). The residue was co-evaporated with EtOAc (2x5 mL) to give compound 16 (163 mg, 85%) as a white solid.

1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 8.57 (d, J=2.0 Hz, 1H), 8.50 (dd, J=4.8, 1.6 Hz, 1H ), 8.50 (dt, J=7.6, 1.6 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.69 (d, J=1.6 Hz , 1H), 7.42 (dd, J=8.0, 2.0 Hz, 1H), 7.42 (ddd, J=7.6, 4.8, 0.8 Hz, 1H), 5, 08 (br.s, 1H), 4.73-4.64 (m, 3H), 4.36-4.26 (m, 3H), 3.81-3.71 (m, 2H), 2.95 (dd, J=15.6, 5.6 Hz, 1H), 2.57 (d, J=15.6 Hz, 1H), 1 ,17 (d, J=6.8 Hz, 3H); LCMS (method A): R _t =7.9 min, m/z calc. for C ₂₃ H ₂₁ Cl ₂ N ₅ O2 469, m/z, The found value is 470 [M+H] ⁺ .

3.2.1.17. Synthesis of compound 17.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(pyridin-4-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrαzolo[1,5-a]pyrαzin-10(7H)-one

Sodium cyanoborohydride (134 mg, 2.02 mmol) was added to a mixture of intermediate I13 (440 mg, 0.67 mmol) and 4-pyridinecarboxaldehyde (108 mg, 1.01 mmol) in MeOH (5 mL). The reaction mixture was stirred at room temperature overnight. Additional 4-pyridinecarboxaldehyde (108 mg, 1.01 mmol) and sodium cyanoborohydride (134 mg, 2.02 mmol) were added and the reaction mixture was stirred overnight. The mixture was concentrated under reduced pressure. The residue was dissolved in THF (3 mL). DBU (0.50 mL, 3.37 mmol) and TBD (28.1 mg, 0.20 mmol) were added. The reaction mixture was stirred at 100 °C for 2 h in a sealed pressure tube and the mixture was concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100). The residue was dissolved in MeOH (3 mL). HCl (1 M aq, 1 mL) was added and the mixture was stirred at room temperature overnight. The solution was concentrated in a sample concentration device. The residue was dissolved in DCM (2 mL) and the solution was washed with Na2CO3 (aq) and dried over an HM-N Isolute cartridge. The filtrate was concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100) to afford compound 17 (48 mg, 15%).

1H NMR (400 MHz, DMSO-d ₆ , 101°C) δ ppm 8.48-8.54 (m, 2H), 7.68 (d, J=8.1 Hz, 1H), 7.65 ( d, J=2.0 Hz, 1H), 7.39 (dd, J=8.1, 2.0 Hz, 1H), 7.27-7.32 (m, 2H), 4.94-5 .17 (m, 1H), 4.67 (s, 2H), 4.23-4.36 (m, 3H), 3.733.78 (m, 2H), 2.89-2.98 (m, 2H ), 2.57 (d, J=15.8 Hz, 1H), 1.17 (d, J=7.0 Hz, 3H); LCMS (method B): Rt=1.72 min, mass/charge calcd. for C ₂₃ H ₂₁ Cl ₂ N ₅ O2 469.1, found value 470.1 [M+H]+.

- 62 047929

3.2.1.18. Synthesis of compound 18.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1-methyl-1H-pyrazol-5-yl)methyl]-1,2,3,4,8,9hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I14 (130 mg, 285 μmol, 83% purity) in DMF (5 mL) was added NaH (60% in mineral oil, 12.5 mg, 0.31 mmol). The reaction mixture was stirred at room temperature for 30 min. To a solution of 5-(chloromethyl)-1-methyl-1H-pyrazole hydrochloride (52.3 mg, 0.31 mmol) in DMF (5 mL) was added NaH (60% in mineral oil, 12.5 mg, 0.31 mmol). The mixture was stirred at room temperature for 10 min and added to a mixture of intermediate I14 and NaH in DMF. The reaction mixture was stirred at room temperature overnight, diluted with H2O (150 mL) and extracted with EtOAc (3 x 150 mL). The combined organic extracts were washed with _NaHCO3 (sat., aq., 3 x 150 mL), brine (150 mL), dried ( _Na2SO4 ), filtered and concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, cyclohexane/EtOAc, gradient from 80:20 to 50:50). _A second purification was accomplished by reversed phase flash column chromatography (C-18, MeCN/H2O, gradient from 1:9 to 1:1). The residue was further purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). The product was co-evaporated with EtOH and dried at 50°C under vacuum overnight to afford compound 18 (83.2 mg, 62%).

¹ H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 7.71 (d, J=8.0 Hz, 1H), 7.69 (d, J=2.1 Hz, 1H), 7.42 (dd, J=8.0, 2.1Hz, 1H), 7.34 (d, J=1.8 Hz, 1H), 6.27 (d, J=1.8 Hz, 1H) , 5.08 (br.s, 1H), 4.83-4.52 (m, 3H), 4.38-4.21 (m, 3H), 3.80 (s, 3H), 3.74 -3.69 (m, 2H), 2.95 (dd, J=16.3, 6.6 Hz, 1H), 2.57 (d, J=16.3 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): R _t =9.1 min, m/z calc. for C ₂₂ H ₂₂ Cl ₂ N ₆ O ₂ 472, mass/charge, found value 473 [M+H]+.

3.2.1.19. Synthesis of compound 19.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1-methyl-1H-pyrazol-3-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin- 10(7H)-one

candidate of technical sciences, o/p 19

Compound 19 was prepared according to the procedure described for the synthesis of compound 18.

The crude mixture was purified by flash column chromatography (silica, EtOAc/MeOH, gradient 99:1 to 92:8). The residue was triturated in _Et2O (3 mL) and successively co-evaporated with EtOAc and a mixture of EtOAc and EtOH (1:1, 4x5 mL) to give compound 19 (92 mg, 43%).

1H NMR (400 MHz, DMSO-de, 80°C) δ ppm 7.71 (d, J=8.0 Hz, 1H), 7.69 (d, J=2.0 Hz, 1H), 7, 58 (d, J=2.4 Hz, 1H), 7.42 (dd, J=8.0, 2.0 Hz, 1H), 6.15 (d, J=2.0 Hz, 1H), 5.06 (br.s, 1H), 4.63 (br.s, 1H), 4.63-4.54 (m, 2H), 4.32-4.22 (m, 3H), 3, 80 (s, 3H), 3.75-3.71 (m, 2H), 2.94 (dd, J=16.0, 5.2 Hz, 1H), 2.57 (d, J=16.0 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): R _t =9.1 min, mass /charge calc. for C ₂₂ H ₂₂ Cl ₂ N ₆ O ₂ 472, mass/charge, found value 473 [M+H]+.

- 63 047929

3.2.1.20. Synthesis of compound 20

Intermediate connection I15.

3-{[(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-10-oxo-1,3,4,7,8,10-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-9(2H)-yl]methyl}-N, ^-dimethyl-III-pyrazole-Gsulfunamide

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I14 (185 mg, 0.49 mmol) in DMF (9 mL) was added NaH (60% in mineral oil, 21.4 mg, 0.54 mmol). The reaction mixture was stirred at room temperature for 30 min. A solution of 3-(chloromethyl)-N-dimethyl-1H-pyrazole-1-sulfonamide (120 mg, 0.54 mmol) in DMF (9 mL) was added and the reaction mixture was stirred at room temperature overnight. Additional 3-(chloromethyl)-N-N-dimethyl-1H-pyrazole-1-sulfonamide (21.4 mg, 0.54 mmol) was added and the reaction mixture was stirred for an additional 3 h. The reaction mixture was diluted with _H2O (45 mL) and extracted with EtOAc (3 x 45 mL). The combined organic layers were washed with _NaHCO3 (sat., aq., 3 x 45 mL) and brine (45 mL), dried ( _{Na2SO4 )} , filtered, and concentrated to dryness. The crude mixture was combined with another fraction (0.11 mmol) and purified by flash column chromatography (silica, EtOAc/MeOH, gradient from 99:1 to 92:8) to give intermediate compound I15 (208 mg, 70%).

Compound 20 (3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1 I-pyrazol-3 -yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4‘,3‘:3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I15 (188 mg, 0.33 mmol) was solubilized in HCl (4N in 1,4-dioxane, 9.13 mL, 36.5 mmol). The reaction mixture was stirred at room temperature for 15 h and quenched with _NaHCO3 (sat., aq., 9 mL). The aqueous phase was extracted with EtOAc (3x25 mL). The combined organic layers were washed with water (25 mL) and brine (25 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/ _MeOH , 96:4). The residue was triturated in _Et2O (3 ml), co-evaporated with EtOAc (2x5 ml) and EtOH (2x5 ml) and dried under vacuum overnight at 55°C to give compound 20 (60 mg, 39%) as a white solid.

^1H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 12.52 (br.s, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.69 (d , J=2.0 Hz, 1H), 7.62 (br.s, 1H), 7.42 (dd, J=8.0, 2.0 Hz, 1H), 6.19 (d, J= 1.6 Hz, 1H), 5.07 (br.s, 1H), 4.79-4.53 (m, 3H), 4.37-4.21 (m, 3H), 3.78-3 .68 (m, 2H), 2.95 (dd, J=16.0, 6.0 Hz, 1H), 2.57 (d, J=16.0 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): Rt=8.8 min, m/z calc. for C ₂₁ H ₂₀ Cl ₂ N ₆ O ₂ 458, mass/charge, found value 459 [M+H]+.

3.2.1.21. Synthesis of compound 21.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1,2-oxazol-3-yl)methyl]-1,2,3,4,8,9-hexahydropyri- 64 047929 do[4',3':3,4]pyrazolo[1,5 -a]pyrazin-10(7H)-one

Compound 21 was prepared according to the procedure described for the synthesis of compound 15.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). A second purification was accomplished by preparative HPLC (XBridge 5 μm OBD column, 30x150 mm, 40 mL/min, _H2O +0.1% HCOOH/MeOH: 54/46). The residue was taken up in a mixture of EtOAc (30 mL) and _NaHCO3 (sat., aq., 30 mL). The layers were separated and the aqueous phase was extracted with _EtOAc (2x25 mL). The combined organic extracts were dried ( _Na2SO4 ), filtered and concentrated under reduced pressure to afford compound 21 (52 mg, 28%) as a white solid.

1H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 8.82 (d, J=1.6 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7 .69 (d, J=2.0 Hz, 1H), 7.42 (dd, J=8.4, 2.0 Hz, 1H), 6.54 (d, J=1.6 Hz, 1H) , 5.06 (br.s, 1H), 4.80-4.72 (m, 2H), 4.60 (br.s, 1H), 4.38-4.26 (m, 3H), 3 .87-3.77 (m, 2H), 2.95 (dd, J=16.0, 5.6 Hz, 1H), 2.58 (d, J=16.0 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): R _t =9.4 min, m/z calc. for C ₂₁ H ₁₉ Cl ₂ N ₅ O ₃ 459, mass/charge, found value 460 [M+H]+.

3.2.1.22. Synthesis of compound 22.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1,3-thiazol-4-yl)methyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5^pyrazin-1 0(7H)-one

Compound 22 was prepared according to the procedure described for the synthesis of compound 15.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 95:5) to give 120 mg of impure compound 22. The fraction was combined with another crop (0.38 mmol) and purified by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 90:10). A second purification was accomplished by flash column chromatography (C-18, _H2O /MeCN, gradient 60:40 to 0:100). The residue was again purified by flash column chromatography (silica, EtOAc) to give compound 22 (50 mg, 15%) as a white solid.

^1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 9.04 (d, J=1.6 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7 .69 (d, J=2.0 Hz, 1H), 7.59 (d, J=1.6 Hz, 1H), 7.42 (dd, J=8.0, 1.6 Hz, 1H) , 5.07 (br.s, 1H), 4.84-4.75 (m, 2H), 4.60 (br.s, 1H), 4.37-4.26 (m, 3H), 3 .88-3.78 (m, 2H), 2.95 (dd, J=16.0, 6.0 Hz, 1H), 2.58 (d, J=16.0 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): Rt=9.3 min, m/z calc. for C ₂₁ H ₁₉ Cl ₂ N ₅ O ₂ S 475, mass/charge, found value 476 [M+H]+.

3.2.1.23. Synthesis of compound 23

Intermediate connection I16.

Ethyl (6R)-5-(3,4-dichlorobenzoyl)-2-(2-{[(1R)-1-(4-methoxyphenyl)ethyl]amino}ethyl)-6-methyl-4,5,6,7tetrahydro^Spyrazolo^, 3-c]pyridine-3-carboxylate

- 65 047929

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I9 (700 mg, 1.39 mmol) in MeCN (25 mL) were added KI (92.2 mg, 0.56 mmol) and ((-)-1-(4-methoxyphenyl)ethylamine (1.0 mL, 6.95 mmol). The reaction mixture was stirred at 90 °C for 20 h and diluted with H2O (150 mL). The aqueous phase was extracted with EtOAc (3 x ₁₅₀ mL). The combined organic extracts were dried ( _Na2SO4 ), filtered and concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5) to give intermediate I16 (523 mg, 67%).

Connection 23.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(1R)-1-(4-methoxyphenyl)ethyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of intermediate I16 (524 mg, 0.94 mmol) in DCE (6 ml) was added AlMe ₃ (2 M in toluene, 1.40 ml, 2.80 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 1 h and at 80 °C for 1 h. Additional AlMe3 (2 M in toluene, 1.40 ml, 2.80 mmol) was added and the reaction mixture was stirred at 80 °C for another h. The reaction mixture was diluted with DCM (150 ml) and Na ₂ SO ₄ -10H ₂ O (1.5 g) was added. The mixture was stirred for 30 min, dried (Na ₂ SO ₄ ) and filtered. The organic layer was washed with HCl (1N, aq., 3x150 ml), dried ( _Na2SO4 ), filtered and evaporated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/ _MeOH , gradient from 100:0 to 95:5) to give compound 23 (340 mg, 71%).

¹ H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.71 (d, J=8.3 Hz, 1H), 7.69 (d, J=2.1 Hz, 1H), 7 .42 (dd, J=8.3, 2.1 Hz, 1H), 7.30 (d, J=8.6 Hz, 2H), 6.93 (d, J=8.6 Hz, 2h) , 5.78 (q, J=7.0 Hz, 1H), 5.07 (br.s, 1H), 4.63 (br.s, 1H), 4.38-4.21 (m, 2H ), 4.19-4.11 (m, 1H), 3.77 (s, 3H), 3.71-3.62 (m, 1H), 3.33-3.26 (m, 1H), 2.95 (dd, J=16.3, 6.6 Hz, 1H), 2.57 (d, J=16.3 Hz, 1h), 1 ,54 (d, J=7.0 Hz, 3H), 1.18 (d, J=6.8 Hz, 3H); LCMS (method A): R _t =10.9 min, m/z calc. for C ₂₆ H ₂₆ Cl ₂ N ₄ O ₃ 512, mass/charge, the found value is 513 [M+H]+.

3.2.1.24. Synthesis of compound 24

Intermediate connection I17.

Ethyl (6R)-5-(3,4-dichlorobenzoyl)-2-(2-{[(1,5)-1-(4-methoxyphenyl)ethyl]amino}ethyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

Intermediate I17 was prepared according to the procedure described for the synthesis of intermediate I16.

- 66 047929

The crude mixture was purified by flash column chromatography (silica, cyclohexane/EtOAc, gradient 90:10 to 0:100). A second purification was achieved by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 95:5) to afford intermediate I17 (631 mg, 81%).

Connection 24.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(1S)-1-(4-methoxyphenyl)ethyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]i sraish-1 0(7H)-one

Compound 24 was prepared according to the procedure described for the synthesis of compound 23.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 95:5) to afford compound 24 (447 mg, 77%, 95% purity).

1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.71 (d, J=8.3 Hz, 1H), 7.69 (d, J=2.1 Hz, 1H), 7, 42 (dd, J=8.3, 2.1 Hz, 1H), 7.28 (d, J=8.6 Hz, 2H), 6.92 (d, J=8.6 Hz, 2H), 5.78 (q, J=7.0 Hz, 1H) 5.11 (br.s, 1H), 4.62 (br.s, 1H), 4.37-4.21 (m, 2H), 4.16-4.05 (m, 1H), 3.76 (s, 3H), 3.70-3.63 (m, 1H), 3.30-3.22 (m, 1H), 2.94 (dd, J=16.3, 6.6 Hz, 1H), 2.56 (d, J=16.3 Hz, 1H), 1 ,54 (d, J=7.0 Hz, 3H), 1.17 (d, J=6.8 Hz, 3H); LCMS (method A): Rt=10.8 min, m/z calc. for C ₂₆ H ₂₆ Cl ₂ N ₄ O ₃ 512, mass/charge, found value 513 [M+H]+.

3.2.1.25. Synthesis of compound 25

Intermediate connection I18.

Ethyl (6R)-5-(3,4-dichlorobenzoyl)-2-(2-{[(1R)-1-(4-fluorophenyl)ethyl]amino}ethyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

Intermediate I18 was prepared according to the procedure described for the synthesis of intermediate I16.

The crude mixture was purified by flash column chromatography (silica, cyclohexane/EtOAc, gradient 90:10 to 0:100) to give intermediate I18 (397 mg, 79%).

Connection 25.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(1R)-1-(4-fluorophenyl)ethyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

Compound 25 was prepared according to the procedure described for the synthesis of compound 23.

The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). Purification was accomplished by preparative SFC (non-sulfuric acid).

- 67 047929 mobile phase: Chiralpak Daicel ID 20x250 mm, mobile phase: CO ₂ , i-PrOH+0.4% i-PrNH ₂ ) to obtain compound 25 (195 mg, 56%).

1H NMR (400 MHz, DMSO-d6) δ ppm 1.16 (d, J=6.8 Hz, 3H), 1.54 (d, J=7.0 Hz, 3H), 2.55 (d, J=16.1 Hz, 1H), 2.87-2.95 (m, 1H), 3.21-3.41 (m, 1H), 3.56-3.74 (m, 1H), 4 .05-4.36 (m, 3H), 4.53 (br s, 1H), 5.05 (br s, 1H), 5.78 (q, J=7.0 Hz, 1H), 7, 09-7.16 (m, 2H), 7.36-7.43 (m, 3H), 7.62-7.71 (m, 2H); LCMS (method D): Rt=2.14 min, m/z calc. for C ₂₅ H ₂₃ Cl ₂ FN ₄ O ₂ , 500.1; m/z found 501.0 [M+H ] ⁺ .

3.2.1.26. Synthesis of compound 26

Intermediate connection I19.

Ethyl (6R)-5-(3,4-dichlorobenzoyl)-2-(2-{[(1S)-1-(4-fluorophenyl)ethyl]amino}ethyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

Intermediate I19 was prepared according to the procedure described for the synthesis of intermediate I16.

The crude mixture was purified by flash column chromatography (silica, cyclohexane/EtOAc, gradient 90:10 to 0:100) to give intermediate I19 (709 mg, 72%).

Connection 26.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(1S)-1-(4-fluorophenyl)ethyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

Compound 26 was prepared according to the procedure described for the synthesis of compound 23.

The crude mixture was purified by flash column chromatography (silica, DCM/EtOAc, gradient from 50:50 to 20:80, then DCM/MeOH, gradient from 100:0 to 95:5). The second purification was accomplished by flash column chromatography (silica, DCM/THF, gradient from 100:0 to 70:30). Finally, the residue was purified by preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO ₂ , EtOH+0.4% i-PrNH ₂ ) to afford compound 26 (80 mg, 26%).

¹ H NMR (400 MHz, DMSO-d6) δ ppm 1.16 (d, J=6.8 Hz, 3H), 1.55 (d, J=7.3 Hz, 3H), 2.56 (d , J=16.1 Hz, 1H), 2.92 (dd, J=16.0, 5.8 Hz, 1H), 3.30 (ddd, J=13.2, 7.0, 4.7 Hz, 1H), 3.69 (ddd, J=13.0, 8.1, 4.6 Hz, 1H), 4.084.17 (m, 1H), 4.21-4.32 (m, 2H) , 4.55-4.74 (m, 1H), 4.94-5.22 (m, 1H), 5.79 (q, J=7.2 Hz, 1H), 7.09-7.16 (m, 2H), 7.35-7.42 (m, 2H), 7.64-7.71 (m, 2H); LCMS (method D): R _t = 2.14 min, mass/charge calculated for C ₂₅ H ₂₃ Cl ₂ FN ₄ O ₂ , 500.1; mass/charge found value 501.2 [M+H] ⁺ .

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3.2.1.27. Synthesis of compound 27

Intermediate connection I20.

(31<)-2-(3,4-Dichlorobenzoyl)-9-[(11<)-1-(4-1'hydro1<cyphenyl)ethyl]-3-methyl-1,2,3,4,8,9hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

To a solution of compound 23 (200 mg, 0.39 mmol) in DCM (4 mL) at -78 °C was added _BBr3 (1 M in DCM, 2.34 mL, 2.34 mmol). The reaction mixture was stirred at -78 °C for 1 h, warmed to -35 °C and stirred for 1 h, warmed to -12 °C and stirred for another h. The reaction mixture was quenched with _NaHCO3 (sat., aq., 15 mL) and the mixture was stirred for 15 min and concentrated under reduced pressure. MeOH (10 mL) was added and the reaction mixture was stirred at room temperature overnight. The aqueous layer was extracted with EtOAc (3 x ₁₀₀ mL). The combined organic layers were dried ( _Na2SO4 ), filtered and concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, DCM/EtOAc, gradient 60:40 to 0:10) to give intermediate I20 (135 mg, 65%, purity 93%).

Connection 27.

(3R)-2-(3,4-Dichlorobenzoyl)-9-{(1R)-1-[4-(difluoromethoxy)phenyl]ethyl}-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out in an argon atmosphere.

To a suspension of intermediate I20 (100 mg, 0.20 mmol) and KOH (225 mg, 4.00 mmol) in _H2O (350 μL) and MeCN (350 μL) at -78°C was added diethyl [(bromodifluoro)methyl]phosphonate (71 μL, 0.40 mmol). The reaction mixture was stirred at room temperature for 1 h and diluted with EtOAc (50 mL). The mixture was washed with _NaHCO3 (sat., aq.), brine, dried (Na2SO4), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 97:3) to afford compound 27 (48 mg, 43%) as a light yellow solid.

Ή NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 7.73 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.6 Hz, 1H), 7 .47-7.41 (m, 3H), 7.18 (d, J=8.4 Hz, 2H), 7.15 (t, J=74.2 Hz, 1H), 5.81 (q, J=7.2 Hz, 1H), 5.05 (br.s, 1H), 4.65 (br.s, 1H), 4.37-4.15 (m, 3H), 3.75-3 .67 (m, 1H), 3.39-3.31 (m, 1H), 2.95 (dd, J=16.0, 6.0 Hz, 1H), 2.58 (d, J=16.0 Hz, 1H), 1.56 (d, J=7.2 Hz, 3H), 1.18 (d, J=6.7 Hz, 3H); LCMS (method A): R _t = 11.1 min, m/z calc. for C ₂₆ H ₂₄ Cl ₂ F ₂ N ₄ O ₃ 548, m/z found 549 [M+H]+ . 3.2.1.28.

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Synthesis of compound 28

121

Cl

Cl

_BrCF2P (=O)(OEt) _2KOH

MeCN: _H2O from -78°C to rt, 1 h.

Intermediate compound I21.

(3R)-2-(3,4-Dichlorobenzoyl)-9-[(SR)-1-(4-hydroxyphenyl)ethyl]-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

Intermediate I21 was prepared according to the procedure described for the synthesis of intermediate I20.

The crude mixture was purified by flash column chromatography (silica, DCM/EtOAc, gradient from 60:40 to 0:100) to give intermediate compound I21 (173 mg, 65%, purity 91%).

Connection 28.

(3R)-2-(3,4-Dichlorobenzoyl)-9-{( 1S)- 1-[4-(difluoromethoxy)phenyl]ethyl}-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

I 1 g G

About Uch/

About VSL/Λ

A °cf _2H

Compound 28 was prepared according to the procedure described for the synthesis of compound 27.

The crude mixture was purified several times (3 times) by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 97:3) to give compound 28 (67 mg, 47%) as a white solid.

^1H NMR (400 MHz, DMSO-d6, 80°C) δ ppm 7.72 (d, J=8.2 Hz, 1H), 7.70 (d, J=1.9 Hz, 1H), 7 .46-7.41 (m, 3H), 7.17 (d, J=8.4 Hz, 2H), 7.14 (t, J=74.1 Hz, 1H), 5.81 (q, J=7.2 Hz, 1H), 5.10 (br.s, 1H), 4.62 (br.s, 1H), 4.37-4.26 (m, 2H), 4.21-4 .13 (m, 1H), 3.77-3.68 (m, 1H), 3.37-3.29 (m, 1H), 2.95 (dd, J=16.0, 5.6 Hz, 1H), 2.58 (d, J=16.0 Hz, 1H), 1.57 (d, J=6.9 Hz, 3H), 1 ,18 (d, J=6.9 Hz, 3H); LCMS (method A): R _t =11.1 min, m/z calc. for C ₂₆ H ₂₄ Cl ₂ F ₂ N ₄ O ₃ 548, m/z /charge, found value 549 [M+H]+. 3.2.1.29.

Synthesis of compound 29

TBD

THF: _H2O microwave, 100°C, 1.8 h.

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Intermediate compound I22.

Ethyl (6k)-5-(3,4-dichlorobenzoyl)-2-[2-({(^)-1-[1-(dimethylsulfamoyl)-III-1,2,4-triazol-3yl]ethyl}amino)ethyl]-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I9 (362 mg, 0.74 mmol) was dissolved in MeCN (12 mL). KI (49.2 mg, 0.30 mmol) and intermediate I3 (820 mg, 3.70 mmol) were added. Additional KI (49.2 mg, 0.30 mmol) was added and the reaction mixture was stirred for another 1.5 h. The reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3 x ₂₀ mL). The combined organic layers were dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). The second purification was accomplished by flash column chromatography (silica, DCM/MeOH, 95:5) to afford a pure fraction of intermediate I22 (292 mg, 61%) and another fraction containing impurities (49 mg, 9%, purity 84%).

Connection 29.

3-{(1S)-1-[(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-10-oxo-1,3,4,7,8,10-hexahydropyrido[4',3 ':3,4]pyrazolo[1,5-a]pyrazin-9(2H)-yl]ethyl}-N,N-dimethyl-1H-1,2,4-triazol-1-sulfonamide

The reaction was carried out in an argon atmosphere.

Intermediate I22 (292 mg, 0.45 mmol) was dissolved in MeCN (1.1 mL) and H2O (5.9 mL). TBD (37.9 mg, 0.27 mmol) was added and the reaction mixture was stirred at 100 °C under microwave irradiation for 1.8 h. The reaction mixture was concentrated to dryness. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 98:2 to 95:5) to give compound 29 (215 mg, 71%, 87% purity).

28 mg of compound 29 were co-evaporated with EtOAc (3x2 ml) and dried under vacuum at 50°C for 20 h to give pure compound 29 (24 mg) as a white solid.

^1H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 9.00 (s, 1H), 7.72 (d, J=8.3 Hz, 1H), 7.69 (d, J =2.1 Hz, 1H), 7.42 (dd, J=8.3, 2.1 Hz, 1H), 5.86 (q, J=6.9 Hz, 1H), 5.06 (br .s, 1H), 4.61 (br.s, 1H), 4.39-4.19 (m, 3H), 3.863.78 (m, 1H), 3.65-3.58 (m, 1H ), 2.95 (dd, J=16.3, 6.6 Hz, 1H), 2.91 (s, 6H), 2.58 (d, J=16.3 Hz, 1H), 1.61 (d, J=6.9 Hz, 3H), 1.17 (d, J=6.8 Hz, 3H); LCMS ( method A): R _t = 10.1 min, mass/charge calc. for C ₂₃ H ₂₆ Cl ₂ N ₈ O ₄ S 580, mass/charge, found value 581 [M+H]+.

3.2.1.30. Synthesis of compound 30.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1 S)- 1-(1H-1,2,4-triazol-3-yl)ethyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Compound 29 (178 mg, 0.27 mmol, 87% purity) was solubilized in 1,4-dioxane (1 mL) and HCl (4 N in 1,4-dioxane, 5 mL, 20 mmol) was added. The reaction mixture was stirred at room temperature for 2 h and concentrated to dryness. The residue was diluted in DCM (5 mL) and washed with _NaHCO3 (sat., aq., 3x5 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). The residue was co-evaporated with EtOH (3x3 ml) and dried under vacuum at 50°C for 48 h to give compound 30 (102 mg, 80%) as a white solid.

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1H NMR (400 MHz, DMSO-d ₆ , 80°C) δ ppm 13.74 (br.s, 1H), 8.24 (s, 1H), 7.71 (d, J=8.3 Hz, 1H), 7.69 (d, J=2.1 Hz, 1H), 7.42 (dd, J=8.3, 2.1 Hz, 1H), 5.86 (q, J=7.2 Hz, 1H), 5.08 (br.s, 1H), 4.62 (br.s, 1H), 4.35-4.15 (m, 3H), 3.81-3.72 (m, 1H), 3.62-3.53 (m, 1H), 2.95 (dd, J=16.3, 6.6 Hz, 1H), 2.57 (d, J=16.3 Hz, 1H), 1.57 (d, J=7.2 Hz, 3H), 1.17 (d, J=6.8 Hz, 3H); LCMS ( method A): R=8.5 min, mass/charge calc. for C21H21CW7O2 473, mass/charge, found value 474 [M+H]+.

3.2.1.31. Synthesis of compounds 31 and 32

TBD

THF: _H2O microwave, 100°C, 1.8 h.

CI

NH

CI

CI

CI

NH

Intermediate compound I23.

Ethyl-(6R)-2-{2-[(but-3-yn-2-yl)amino]ethyl}-5-(3,4-dichlorobenzoyl)-6-methyl-4,5,6,7- tetrahydro-2Hpyrazolo[4,3-c]pyridine-3-carboxylate

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I9 (214 mg, 0.42 mmol) was dissolved in MeCN (7.60 mL). KI (28.2 mg, 0.17 mmol) and 1-methylprop-2-ynylamine (179 µL, 2.12 mmol) were added. The reaction mixture was stirred at 90 °C for 24 h. Additional KI (28.2 mg, 0.17 mmol) was added and the reaction mixture was stirred at 90 °C for 17 h. 1-Methylprop-2-ynylamine (71.5 µL, 0.85 mmol) was added and the reaction mixture was stirred at 90 °C for an additional 2 h. The reaction mixture was transferred to a sealed tube, 1-methylprop-2-ynylamine (179 µL, 2.12 mmol) was added and the reaction mixture was stirred at 90 °C for 16 h. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient 100:0 to 95:5) to afford intermediate I23 (177 mg, 78%, 89% purity) as a yellowish gum.

Intermediate connection I24.

(3R)-9-(But-3-yn-2-yl)-2-(3,4-dichlorobenzoyl)-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5w]pyrazin-1 0(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I23 (177 mg, 0.33 mmol, 89% purity) was dissolved in MeCN (0.6 mL) and THF (2.92 mL). TBD (13.8 mg, 0.10 mmol) was added and the reaction mixture was stirred at 100 °C under microwave irradiation for 1.8 h. Additional TBD (13.8

- 72 047929 mg, 0.10 mmol) and the reaction mixture was stirred at 100 °C under microwave irradiation for an additional 1.8 h. The reaction mixture was concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5) to give intermediate compound I24 (156 mg, 84%, purity 77%).

Compounds 31 and 32.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1*R)-1-(1H-1,2,3-triazol-4-yl)ethyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1 ^^pyrazin-1 0(7H)-one (3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-9-[(1*S)-1-(1H-1,2,3-triazol-4-yl)ethyl]-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1 ^^pyrazin-1 0(7H)-one

The reaction was carried out under anhydrous conditions and in an argon atmosphere.

Intermediate I24 (126 mg, 0.22 mmol, 77% purity) was dissolved in DMF (1.35 mL) and MeOH (150 μL). CuI (2.14 mg, 0.01 mmol) and trimethylsilyl azide (44.3 μL, 0.34 mmol) were added. The reaction mixture was stirred at 100 °C for 16 h. The mixture was combined with another fraction (0.05 mmol) and concentrated to dryness. The residue was diluted with EtOAc (5 mL) and washed with water (3 x 5 mL). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, DCM/MeOH, gradient from 100:0 to 95:5). The residue was co-evaporated with EtOAc (3x2 mL) and dried under vacuum at 50°C to give compound I25 (77 mg, 72%) as a white solid.

Diastereoisomers were separated by preparative SFC (stationary phase: Chiralpak Daicel ID 20x250 mm, mobile phase: CO ₂ , EtOH+0.4% i-PrNH ₂ ) to afford compounds 31 and 32.

Connection 31.

1H NMR (400 MHz, DMSO-d ₆ , 81°C) δ ppm 14.10-14.97 (m, 1H), 7.80 (s, 1H), 7.66-7.72 (m, 2H ), 7.41 (dd, J=8.1, 2.0 Hz, 1H), 5.89 (q, J=7.0 Hz, 1H), 4.87-5.25 (m, 1H) , 4.44-4.83 (m, 1H), 4.15-4.35 (m, 3H), 3.663.75 (m, 1H), 3.40-3.46 (m, 1H), 2 .93 (dd, J=16.0, 5.8 Hz, 1H), 2.56 (d, J=15.8 Hz, 1H), 1.55 (d, J=7.0 Hz, 3H), 1.16 (d, J=6.8 Hz, 3H); LCMS (method D): R _t =1.65 min, mass/charge calc. for C ₂₁ H ₂₁ Cl ₂ N ₇ O ₂ 473.1, mass/charge, found value 474.2 [M+H]+.

Connection 32.

1H NMR (400 MHz, DMSO-d ₆ , 81°C) δ ppm 14.45-14.80 (m, 1H), 7.78 (s, 1H), 7.66-7.72 (m, 2H ), 7.41 (dd, J=8.1, 2.0 Hz, 1H), 5.89 (q, J=7.0 Hz, 1H), 4.86-5.30 (m, 1H) , 4.42-4.83 (m, 1H), 4.23-4.32 (m, 2H), 4.134.21 (m, 1H), 3.67-3.76 (m, 1H), 3 .37-3.49 (m, 1H), 2.93 (dd, J=15.8, 5.7 Hz, 1H), 2.56 (d, J=15.8 Hz, 1H), 1.56 (d, J=7.0 Hz, 3H), 1.16 (d, J=6.8 Hz, 3H); LCMS (method D): R _t = 1.65 min, m/z calc. for C ₂₁ H ₂₁ Cl ₂ N ₇ O2 473.1, m/z found 474.2 [M+H] ⁺ .

3.2 Synthesis of 7-membered rings.

3.2.1. Synthesis of intermediate compound I26.

Ethyl 2-{3-[(tert-butoxycarbonyl)amino]propyl}-5-(3,4-dichlorobenzoyl)-4,5,6,7-tetrahydro-2Hpyrαzolo[4,3-c]pyridine-3-carboxylate

114 I26

A mixture of intermediate I14 (1.00 g, 2.70 mmol), _Cs2CO3 (1.32 g, 4.05 mmol), and 3-(boc-amino)propyl bromide (676 mg, 2.84 mmol) in MeCN (30 _mL ) was stirred at room temperature for 2 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 90:10 to 0:100) to give intermediate I26 (850 mg, 60%).

3.2.2. Synthesis of compounds.

3.2.2.1. Synthesis of compound 33

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Intermediate connection I27.

1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one hydrochloride

A pressure tube was charged with tert-butyl 11-oxo-3,4,7,8,9,10-hexahydro-1H-pyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (250 mg, 0.78 mmol) in i-PrOH (5 mL). HCl (6 M in i-PrOH, 1.3 mL, 7.80 mmol) was added and the reaction mixture was stirred at 100 °C for 30 min. The mixture was concentrated under reduced pressure. The residue was triturated in DIPE. The solids were collected by filtration and dried to give intermediate I27 (200 mg, quant.) as a white solid.

Connection 33.

2-(3,4-Dichlorobenzoyl)-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin11-one

A pressure tube was charged with intermediate I27 (198 mg, 0.82 mmol), 3,4-dichlorobenzoyl chloride (185 mg, 0.86 mmol), DCM (2.6 mL), water (2.61 mL), and _Na2CO3 (432 mg, 4.08 mmol). The reaction mixture was stirred at room temperature for 16 _h . The precipitate was collected by filtration and washed with water and DIPE. The product was dried under vacuum to give compound 33 (220 mg, 71%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6, 100°C) δ ppm 7.72 (br s, 1H), 7.66 (d, J=8.1 Hz, 1H), 7.63 (d, J=2.0 Hz, 1H), 7.39 (dd, J=8.1, 2.0 Hz, 1H), 4.63 (s, 2H), 4.34 (t, J=6.4 Hz, 2H), 3.62-3.78 (m, 2H), 3.19-3.27 (br s, 2H), 2.70 (t, J=5.8 Hz, 2H), 2.09-2.16 (m, 2H); LCMS (method C): R _t = 0.79 min, m/z calc. for C17H16Cl ₂ N4O ₂ 378.0, the found value is 379.1 [M+H]+; m.p. 237°C.

3.2.2.2. Synthesis of compound 34.

2-(3,4-Dichlorobenzoyl)-10-methyl-1,2,3,4,7,8,9,10-octahydro-1 1Н-pyrbdo[4',3':3,4]pyrazolo[1,5a][1,4]giazenin-11-one

Compound 33 (60.0 mg, 158 μmol), MeI (19.7 μL, 0.32 mmol) and _Cs2CO3 (103 mg, 0.32 mmol) were measured into _a microwave vial. THF (2 mL) was added and the reaction mixture was stirred at 100 °C for 16 h. The mixture was concentrated under reduced pressure. The residue was partitioned between water and 2-MeTHF. The layers were separated and the organic phase was dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 100:0 to 0:100). The product was triturated in DIPE and MeCN. The solids were collected by filtration and dried under vacuum to afford compound 34 (29 mg, 47%) as a pale yellow solid.

¹ H NMR (400 MHz, DMSO-d6, 100°C) δ ppm 7.67 (d, J=8.1 Hz, 1H), 7.64 (d, J=2.0 Hz, 1H), 7.40 (dd, J=8.4, 2.0 Hz, 1H), 4.60 (s, 2H), 4.28 (t, J=6.8 Hz, 2H), 3.63-3.77 (m, 2H), 3.36-3.41 (m, 2H), 3.00 (s, 3H), 2.70 (t, J=5.9 Hz, 2H), 2.16-2.23 (m, 2H); LCMS (method C): Rt=0.83 min, m/z calcd. for Ci8Hi8C1 ₂ N4O ₂ 392.1, the found value is 393.1 [M+H] ⁺ .

3.2.2.4. Synthesis of compound 36.

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2-(3,4-Dichlorobenzoyl)-10-[(4-methoxyphenyl)methyl]-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3 ,4]pyrazolo[1,5-a][1,4]diazepin-11-one

Compound 36 was prepared according to the procedure described for the synthesis of compound 34.

The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient 100:0 to 0:100). The product was triturated in DIPE. The solids were collected by filtration and dried under vacuum to give compound 36 (31 mg, 39%) as a white solid.

¹ H NMR (400 MHz, DMSO-d6, 100°C) δ ppm 7.67 (d, J=8.1 Hz, 1H), 7.64 (d, J=2.0 Hz, 1H), 7.40 (dd, J=8.3, 1.9 Hz, 1H), 7.22-7.28 (m, 2H), 6.86-6.92 (m, 2H), 4.64 (s, 2H), 4.57 (s, 2H), 4.24 (t, J=6.8 Hz, 2H), 3.75 (s, 3H), 3.67-3.74 (m, 2H), 3.31-3.37 (m, 2H), 2.71 (t, J=5.8 Hz, 2H), 1.97-2.05 (m, 2H); LCMS (method C): R _t = 1.04 min, m/z calcd for C ₂₅ H ₂₄ Cl ₂ N ₄ O ₃ 498.1, found 499.1 [M+H]+; mp 236°C.

3.2.2.5. Synthesis of compound 37

1) || u ^ch o

NaBH ₃ CN MeOH grade, o/p

2) TBD DBU THF 100°С, o/p

Intermediate connection I28.

Ethyl 2-(3-aminopropyl)-5-(3,4-dichlorobenzoyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3carboxylate · 2TFA

128

TFA (1.24 ml, 16.2 mmol) was added to a solution of intermediate I26 (850 mg, 1.62 mmol) in DCM (100 ml). The reaction mixture was stirred overnight at room temperature and concentrated under reduced pressure to give intermediate I28, which was used as such in the next step.

Compound 37.

2-(3,4-Dichlorobenzoyl)-10-[(pyridin-4-yl)methyl]-1,2,3,4,7,8,9,10-octahydro-11 H-pyrido[4',3':3,4]pyrazolo[1,5-a] [1,4]diazepin-11 -one

Sodium cyanoborohydride (56.6 mg, 0.86 mmol) was added to a mixture of intermediate I28 (373 mg, 0.57 mmol) and 4-pyridinecarboxaldehyde (61.2 mg, 0.57 mmol) in MeOH (10 mL). The reaction mixture was stirred at room temperature for 4 h. Additional 4-pyridinecarboxaldehyde (61.2 mg, 0.57 mmol) and sodium cyanoborohydride (56.6 mg, 0.86 mmol) were added. The reaction mixture was stirred overnight and concentrated under reduced pressure.

The residue was dissolved in THF (3 mL). TBD (23.8 mg, 0.17 mmol) and DBU (0.43 mL, 2.85

- 75,047,929 mmol) and the reaction mixture was stirred at 100°C overnight in a pressure tube. The mixture was concentrated under reduced pressure and the crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100) to afford compound 37 (99 mg, 37%).

¹ H NMR (400 MHz, DMSO-d ₆ , 101 °C) δ ppm 8.47-8.54 (m, 2H), 7.67 (d, J = 8.1 Hz, 1H), 7.64 (d, J = 1.8 Hz, 1H), 7.40 (dd, J = 8.3, 1.9 Hz, 1H), 7.27 (d, J = 5.7 Hz, 2H), 4.68 (s, 2H), 4.64 (s, 2H), 4.32 (t, J = 6.7 Hz, 2H), 3.64-3.78 (m, 2H), 3.43 (dd, J = 6.8, 5.5 Hz, 2H), 2.73 (t, J = 5.9 Hz, 2H), 2.09-2.17 (m, 2H); LCMS (method B): Rt=1.66 min, m/z calcd for C ₂₃ H ₂₁ Cl ₂ N ₅ O ₂ 469.0, found value 470.2 [M+H]+.

3.2.2.6. Synthesis of compound 38 o

129 38

Intermediate connection I29.

(3R)-3-methyl-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one hydrochloride

A pressure tube was charged with tert-butyl (3R)-3-methyl-11-oxo-3,4,7,8,9,10hexahydro-1H-pyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (250 mg, 0.78 mmol) in i-PrOH (5 mL). HCl (6 M in i-PrOH, 1.3 mL, 7.80 mmol) was added and the reaction mixture was stirred at 100 °C for 30 min. The mixture was concentrated under reduced pressure. The residue was triturated in DIPE. The solids were collected by filtration and dried to give intermediate I29 (200 mg, quant.) as a white solid.

Connection 38.

(3H)-2-(3,4-Dichlorobenzoyl)-3-methyl-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4] pyrazolo[1,5a][1,4]diazepin-11-one

A pressure tube was charged with intermediate I29 (200 mg, 0.78 mmol), 3,4-dichlorobenzoyl chloride (177 mg, 0.82 mmol), DCM (2.5 mL, 39 mmol), and water (2.5 mL). _Na2CO3 (413 mg, 3.90 mmol) was _added portionwise and the reaction mixture was stirred at room temperature for 1 h. The layers were separated and the organic phase was dried ( _MgSO4 ), filtered, and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 100:0 to 0:100). The product was triturated in DIPE, collected by filtration and dried under vacuum to afford compound 38 (172 mg, 56%) as a white friable solid.

^1H NMR (400 MHz, DMSO-d ₆ , 100°C) δ ppm 7.74 (br s, 1H), 7.66 (d, J=8.4 Hz, 1H), 7.62 (d, J=1.8 Hz, 1H), 7.37 (dd, J=8.3, 1.9 Hz, 1H), 4.87-5.12 (m, 1H), 4.47-4.74 (m, 1H), 4.29-4.41 (m, 2H), 4.20 (br d, J=17.4 Hz, 1H), 3.21-3.28 (m, 2H), 2 .94 (s, 1H), 2.90 (dd, J=15.8, 5.9 Hz, 1H), 2.52 (d, J=15.8 Hz, 1H), 2.09-2.18 (m, 2H), 1.14 (d, J=6.8 Hz, 3H); LCMS (method C): Rt=0.83 min, mass/charge calc. for C^C^N^ 392.1, found value 393.2 [M+H] ⁺ .

3.2.2.7. Synthesis of compound 39.

(3R)-2-(3,4-Dichlorobenzoyl)-3-methyl-10-(2-methylpropyl)-1,2,3,4,7,8,9,10-octahydro-1 SH-pyrido[4',3':3 ,4]pyrazolo[1,5-a] [1,4]diazepin-11-one

- 76 047929

Compound 39 was prepared according to the procedure described for the synthesis of compound 34.

The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc, gradient 100:0 to 0:100) to afford compound 39 (45 mg, 66%).

¹ H NMR (400 MHz, CDCl ₃ , 57°C) δ ppm 7.43-7.54 (m, 2H), 7.23 (dd, J=8.1, 1.6 Hz, 1H), 5 .28 (s, 1H), 4.56-5.55 (m, 1H), 4.28-4.48 (m, 3H), 3.32-3.53 (m, 3H), 3.18 -3.30 (m, 1H), 2.91-3.07 (m, 1H), 2.62 (br d, J=15.9 Hz, 1H), 2.21-2.32 (m, 2H), 1.90-2.05 (m, 1H), 1.25 (br d, J=6.9 Hz, 3H), 0.92-1.01 (m, 6H); LCMS (method D): R _t = 2.02 min, m/z calc. for C ₂₂ H ₂₆ Cl ₂ N ₄ O ₂ 448.1, found value 449 ,3 [M+H]+.

3.2.2.8. Synthesis of compound 40.

(3R)-2-(3,4-Dichlorobenzoyl)-10-[(4-methoxyphenyl)methyl]-3-methyl-1,2,3,4,7,8,9,10-octahydro-11Hpyrido [4 ',3':3,4] pyrazolo [1,5-a][1,4] diazepin-11-one

Compound 40 was prepared according to the procedure described for the synthesis of compound 34.

The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 100:0 to 0:100). Further purification was achieved by preparative SFC (stationary phase: Chiralcel Diacel OJ 20 x 250 mm, mobile phase: CO2, EtOH+0.4% i- _PrNH2 ) to afford compound 40 (38 mg, 29%).

^1H NMR (400 MHz, DMSO-d6, 100°C) δ ppm 7.67 (d, J=8.1 Hz, 1H), 7.64 (d, J=2.0 Hz, 1H), 7 .38 (dd, J=8.3, 1.9 Hz, 1H), 7.24-7.29 (m, 2H), 6.86-6.92 (m, 2H), 4.87-5 .13 (m, 1H), 4.52-4.71 (m, 3H), 4.18-4.29 (m, 3H), 3.75 (s, 3H), 3.31-3.39 (m, 2H), 2.91 (dd, J=16.2, 6.1 Hz, 1H), 2.52 (d, J=16.1 Hz, 1H), 1.97-2.05 (m, 2H), 1.16 (d, J=6.8 Hz, 3H); LCMS (method D): R _t =2, 04 min, mass/charge calc. for C ₂₆ H ₂₆ Cl ₂ N ₄ O ₃ 512.1, found value 513.3 [M+H]+.

3.2.2.9. Synthesis of compound 41

[1709843-86-6] I30 131

Et ₃ N

DCM

K.T., o/p

Intermediate connection I30.

5-tert-Butyl-3-ethyl-2-{[1-(methanesulfonyl)aziridin-2-yl]methyl}-2,4,6,7-tetrahydro-5Hpyrazolo[4,3-c]pyridine-3,5-dicarboxylate

- 77 047929

A mixture of 5-tert-butyl 3-ethyl-3,7-dihydro-2H-pyrazolo[4,3-c]pyridine-3,5(4H)-dicarboxylate (2.55 g, 8.63 mmol), _{epibromohydrin (5.00 g, 23.4 mmol) and K2CO3 (} 3.58 g, 25.9 mmol) in MeCN (75 mL) was stirred at reflux overnight. The reaction mixture was filtered through dicalite while still hot. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (silica, heptane/EtOAc, gradient 80:20 to 0:100) to give intermediate I30 (850 mg, 23%).

Intermediate connection 31.

tert-Butyl 8-[(methanesulfonyl)amino]-10-[(4-methoxyphenyl)methyl]-11-oxo-1,3,4,7,8,9,10,11octahydro-2H-pyrido[4',3':3,4]pyrαzolo[1,5-a][1,4]diazepine-2-carboxylate

Ν _Λ NHSO ₂ Me g- ^{2 Vos} O M _k )-OMe

131

A mixture of intermediate I30 (600 mg, 1.40 mmol), 4-methoxybenzyl bromide (192 mg, 1.40 mmol) in MeCN (5 mL) was stirred at 120 °C for 30 min. The reaction mixture was concentrated under reduced pressure and the crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100). The residue (275 mg) was dissolved in THF (2 mL) and TBD (58.5 mg, 0.42 mmol) was added. The reaction mixture was stirred at 100°C in a sealed tube for 17 h. The mixture was concentrated under reduced pressure and the crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 80:20 to 0:100) to give intermediate compound I31 (86 mg, 12%) as a clear oil.

Connection 41.

N - {2-(3,4-Dichlorobenzoyl)-10-[(4-methoxyphenyl)methyl]-11 -oxo-1,3,4,7,8,9,10,11 -octahydro-2Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diαzepin-8-yl}methanesulfonamide

TFA (0.13 ml, 1.66 mmol) was added to the solution of intermediate I31.

(86.0 mg, 0.17 mmol) in DCM (10 mL) and the reaction mixture was stirred at room temperature for 10 min. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM (20 mL). Et ₃ N (92 μL, 0.66 mmol) was added followed by 3,4-dichlorobenzoyl chloride (36.4 mg, 0.17 mmol) and the reaction mixture was stirred at room temperature overnight. The mixture was washed with water, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAC/EtOH, 3:1), gradient from 80:20 to 0:100). The residue was dissolved in MeOH (5 mL) and water was added dropwise until the solution remained cloudy. The mixture was stirred for 2 h. The precipitate was collected by filtration and dried under vacuum at 50 °C to give compound 41 (61 mg, 62%).

^1H NMR (400 MHz, DMSO-d6, 150°C) δ ppm 7.63 (d, J=8.1 Hz, 1H), 7.59 (d, J=1.8 Hz, 1H), 7 .36 (dd, J=8.1, 2.0 Hz, 1H), 7.21-7.27 (m, 2H), 6.86-6.91 (m, 2H), 6.81 (br s, 1H), 4.81 (d, J=14.5 Hz, 1H), 4.58-4.69 (m, 2H), 4.39-4.49 (m, 2H), 4.24 -4.31 (m, 1H), 3.91-4.01 (m, 1H), 3.76 (s, 3H), 3.70-3.75 (m, 2H), 3.40-3.48 (m, 1H), 3.26-3.34 (m, 1H), 2.90 (s, 3H), 2, 74 (t, J=5.8 Hz, 2H); LCMS (method B): Rt=1.84 min, m/z calc. for C ₂₆ H ₂₇ Cl ₂ N ₅ O ₅ S 591.1, found value 592.1 [M+H]+.

3.2.2.10. Synthesis of compound 42.

a) Synthesis of intermediate compound I34

_MeNH2

S o

THF

70°C, 16 hours.

Vos ₂ 0 NaHCO ₃ he ---------—»1,4-dioxane:H3O

30°C, 16 hours.

Sun .N

HE

HCI

DCM

k.t., 30 min.

•OH . HCI

[548482-49-1] I32 I33

I34

Intermediate connection I32.

2-((Methylamino)methyl)prop-2-en-1-ol

I32

A mixture of 5-methylene-1,3,2-dioxathiane-2-oxide (20.0 g, 149 mmol) in methanamine (2 M in THF, 223 mL) in a sealed tube was stirred at 70 °C for 16 h. The mixture was filtered and the

- 78 047929 rat was concentrated under reduced pressure to give intermediate compound I32 as a yellow liquid, which was used as such in the next step.

Intermediate connection I33.

tert-Butyl (2-(hydroxymethyl)allyl)(methyl)carbamate

To a solution of intermediate I32 in 1,4-dioxane (60 mL) and _H2O (60 mL) were added _Boc2O (22.7 g, 104 mmol) and _NaHCO3 (12.5 g, 148 mmol). The reaction mixture was stirred at 30°C for

The mixture was diluted with EtOAc (400 mL) and washed with brine (3 x 80 mL). The organic layer was dried (Na ₂ SO _{4 )} , filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient from 20:1 to 1:1) to give intermediate I33 (12.5 g, 42% over 2 steps) as a light yellow liquid.

Intermediate connection I34.

2-((methylamino)methyl)prop-2-en-1-ol hydrochloride N II · ^НС|

134

To a solution of intermediate I33 (9.50 g, 47.2 mmol) in DCM (20 mL) was added HCl (4 M in 1,4-dioxane, 30 mL) at 0 °C. The reaction mixture was stirred at 20 °C for 30 min and concentrated under reduced pressure to give intermediate I34 as a yellow oil, which was used in the next step without further purification.

b) Synthesis of compound 42

Intermediate connection I35.

(R)-tert-Butyl-3-((2-(hydroxymethyl)allyl)(methyl)carbamoyl)-6-methyl-6,7-dihydro-2Hpyrazolo[4,3-c]pyridine-5(4H)-carboxylate

To a mixture of (R)-5-(tert-butoxycarbonyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (4.09 g, 14.5 mmol) and intermediate I34 (2.0 g) in pyridine (15 mL) was added EDCI (4.18 g, 21.8 mmol). The reaction mixture was stirred at 40 °C for 16 h and diluted with EtOAc (400 mL). The resulting solution was washed with HCl (1N, aq.) (4 x 30 mL), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 1:1 to 0:1). The residue was triturated with EtOAc and petroleum ether (1:1, 10 mL) to give intermediate I35 (2.2 g, 35%, 85% purity) as a light yellow solid.

Intermediate connection I36.

^)-tert-Butyl-3,10-dimethyl-8-methylene-11 -oxo-3,4,8,9,10,11 -hexahydro-1H- 79 047929 pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

Vos

136

To a mixture of intermediate I35 (1.60 g, 3.73 mmol) in THF (2 mL) was added ADDP (1.88 g, 7.46 mmol) and tributylphosphine (1.51 g, 7.46 mmol) under N2. The reaction mixture was stirred at 70 °C for 12 h and diluted with water (100 mL). The layers were separated and the aqueous phase was extracted with EtOAc (2 x 150 mL). The combined organic extracts were washed with brine (40 mL), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 5:1 to 0:1) to give intermediate I36 (1.7 g, 61%, 94% purity) as a white solid.

Intermediate connection I37.

(3R)-3,10-Dimethyl-8-methylidene-1,2,3,4,7,8,9,10-octahydro-1 1H-pyrido[4',3':3,4]pyrazolo[1 .5

a] [ 1,4]diazepin-11 -one -TFA

A nn 1 lAv ^N 'Me -TFA t J o ν' _N H 137

A sealed tube was charged with intermediate I36 (108 mg, 0.31 mmol) and DCM (1 mL). TFA (0.24 mL, 3.13 mmol) was added and the reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to give intermediate I37, which was used as such in the next step.

Connection 42.

(3R)-2-(3,4-Dichlorobenzoyl)-3,10-dimethyl-8-methylidene-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one o ₀ -Ch

Me 42

Intermediate I37 was dissolved in DCM (1 mL) and water (1 mL). 3,4-Dichlorobenzoyl _chloride (70.9 mg, 0.33 mmol) was added followed by _Na2CO3 (166 mg, 1.56 mmol) portionwise over 10 min. The reaction mixture was stirred at room temperature for 1 h. The layers were separated and the organic phase was dried ( _MgSO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc). The second purification was accomplished by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 μm, 50x150 mm, mobile phase: NH4HCO3 (0.25% in Η ₂ Ο)/ΜόΟΗ). The residue was co-evaporated with MeOH and dried under vacuum to afford compound 42 (72 mg, 53% over 2 steps) as a friable white solid.

¹ H NMR (400 MHz, DMSO-d ₆ , 100°C) δ ppm 7.67 (d, J=8.4 Hz, 1H), 7.63 (d, J=2.0 Hz, 1H), 7.38 (dd, J=8.1, 2.0 Hz, 1H), 5.12-5.20 (m, 2h), 4.97 (s, 2H), 4.55-4.70 ( m, 1H), 4.21 (br d, J=17.2 Hz, 1H), 3.95 (s, 2H), 3.01 (s, 3H), 2.87-2.93 (m, 1H), 2.53 (d, J=16.1 Hz, 1H), 1.15 (d, J=6.8 Hz, 3H); LCMS (method C): Rt=0.95 min, mass/charge calcd for _{C20H20Cl2N4O2 418.2} , found value 419.2 [ _{M +} H] ₊ .

3.2.2.11. Synthesis of compound 43

- 80 047929

Intermediate connection I38.

5-tert-Butyl-3-ethyl-2-(2-(chloromethyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-3,5(4H)dicarboxylate

A mixture of 5-tert-butyl 3-ethyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-3,5(4H)-dicarboxylate (60.0 g, 203 mmol), 3-chloro-2-(chloromethyl)prop-1-ene (50.8 g, 406 mmol) and _Cs2CO3 (99.3 g, 305 mmol) in DMF (600 mL) was degassed and purged with N2 (3 times). The reaction mixture was stirred at 50 °C for ₁₆ h under N2. The mixture was poured into water (3 L) and stirred for 5 min. The aqueous phase was extracted with EtOAc (3 x 1 L). The combined organic extracts were washed with brine (3 L), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/ _EtOAc , gradient from 100:1 to 10:1) to give intermediate I38 (30.00 g, 38%) as a yellow oil.

Intermediate connection I39.

5-tert-Butyl-3-ethyl-2-(2-(((2,2,2-trifluoroethyl)amino)methyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3c]pyridine-3,5(4H)-dicarboxylate

G~ ^CF3

Vos

139

A solution of intermediate I38 (500 mg, 1.30 mmol) and 2,2,2-trifluoroethanamine (3.86 g, 39.0 mmol) in EtOH (50.00 mL) was stirred at 90 °C for 32 h. The mixture was diluted with EtOAc (120 mL) and washed with NaHCO ₃ (sat., aq., 120 mL). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica) to give intermediate I39 (460 mg, 79%) as a yellow oil.

Intermediate connection I40.

tert-Butyl-8-methylene-11-oxo-10-(2,2,2-trifluoroethyl)-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

- 81 047929

Vos

140

To a solution of intermediate I39 (460.0 mg, 1.03 mmol) in toluene (10.00 mL) was added Al(Me) ₃ (2 M in toluene, 2.06 mL) at -30 °C. The reaction mixture was stirred at 60 °C for 16 h. The reaction mixture was quenched with _H2O (10.00 mL). _Na2CO3 ( _109.2 mg, 1.03 mmol) and _Boc2O (270 mg, 1.24 mmol) were added and the reaction mixture was stirred at 20 °C for 16 h. The mixture was diluted with EtOAc (50 mL) and washed with HCl (1 M, aq, 50 mL). _The organic phase was dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica) to give intermediate I40 (390 mg, 95%) as a yellow solid.

Intermediate connection I41.

Hydrochloride

8-methylidene-10-(2,2,2-trifluoroethyl)-1,2,3,4,7,8,9,10-octahydro-11H-pyri to[4',3':3,4]pyrazolo [1,5-a][1,4]diazepin-11-one

Intermediate I40 (214 mg, 0.51 mmol) and i-PrOH (3.26 mL) were charged into a sealed tube. HCl (6 M in i-PrOH, 0.85 mL, 5.10 mmol) was added and the reaction mixture was stirred at 100 °C for 30 min. The mixture was concentrated under reduced pressure. The residue was triturated in DIPE, filtered and dried to give intermediate I41 (169 mg, 95%) as a white solid.

Connection 43.

2-(3,4-Dichlorobenzoyl)-8-methylidene-10-(2,2,2-trifluoroethyl)-1,2,3,4,7,8,9,10-octahydro-1 Sh-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one

_Intermediate I41 (169 mg, 0.48 mmol), 3,4-dichlorobenzoyl chloride (109 mg, 0.51 mmol), DCM (1.5 mL), and water (1.5 mL) were charged into a sealed tube. _Na2CO3 (255 mg, 2.41 mmol) was added portionwise and the reaction mixture was stirred at room temperature for 16 h. The layers were separated and the organic phase was dried (MgSO4), filtered, and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/EtOAc, gradient 100:0 to 0:100) to afford compound 43 (183 mg, 78%).

¹ H NMR (400 MHz, DMSO-d ₆ , 100°C) δ ppm 8.66 (dd, J=4.8, 1.5 Hz, 1H), 8.44 (d, J=2.4 Hz , 1H), 7.697.73 (m, 1H), 7.65-7.69 (m, 2H), 7.56 (dd, J=8.0, 4.7 Hz, 1H), 7.41 ( dd, J=8.1, 2.0 Hz, 1H), 7.12-7.18 (m, 2H), 6.78-6.83 (m, 2H), 6.42 (br s, 1H ), 4.34 (br d, J=2.4 Hz, 2H), 4.29 (s, 2H), 3.72 (s, 3H), 3.58-3.66 (m, 2H), 2.41 (t, J=5.8 Hz, 2H); LCMS (method C): R=1.04 min, mass/ charge calculated for C ₂₇ H ₂₃ Cl ₂ N ₅ O ₃ 535.1, found value 536.1 [M+H]+.

3.2.2.12. Synthesis of compound 44

- 82 047929

ci

Cl

Me

Intermediate connection I42.

tert-Butyl-(3K)-3,10-dimethyl-8,11-dioxo-1,3,4,7,8,9,10,11-octahydro-2H-pyrido[4',3':3,4]pyrazolo[1,5-a] [1,4]diazepine-2-carboxylate

To a mixture of intermediate I37 (1.30 g, 3.49 mmol) in THF (60 mL) and H2O (20 mL) were added _OsO4 (177 mg, 698 μmol) and NaIO4 (2.24 g, 10.5 mmol) at 0 °C. The reaction mixture was stirred at 10 °C for 16 h. The mixture was combined with another fraction (volume 2 g) and diluted with EtOAc (100 mL) _{and Na2SO3} (sat., aq., 100 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL). The combined organic extracts were washed with brine (20 mL), dried ( _{Na2SO4 )} , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 1:1 to 0:1) to give intermediate I42 (2.9 g, 70% purity) as a yellow solid.

Intermediate compounds I43 and I44.

tert-Butyl (3R,8S)-8-hydroxy-3,10-dimethyl-11-oxo-1,3,4,7,8,9,10,11-octahydro-2H-pyrido [4',3‘: 3 ,4]pyrazolo[1,5-a] [ 1,4]diazepine-2-carboxylate tert-Butyl (3R,8S)-8-hydroxy-3,10-dimethyl-11-oxo-1,3,4,7,8,9,10,11-octahydro-2H-pyrido [4',3‘: 3,4]pyrazolo[1,5-a] [ 1,4]diazepine-2-carboxylate

To a mixture of intermediate I42 (2.9 g, 70% purity) in MeOH (30 mL) was added _NaBH4 (551 mg, 14.6 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h and the reaction mixture was quenched with H2O (80 mL) at 0 °C. The mixture was extracted with _EtOAc (3 x 80 mL). The combined organic layers were washed with brine (40 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The residue was combined with another crop (200 mg) and purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 1:1 to 0:1) to give a mixture of intermediates I43 and I44 (1.95 g, 89% purity) as a yellow oil.

- 83 047929

Diastereoisomers were separated by SFC (column: AD (250 mm x 30 mm, 5 μm); mobile phase: _CO2 /i-PrOH (with 1% _NH3 in _H2O ), isocratic elution: 80:20) to afford intermediate I43 and intermediate I44.

SFC analysis: Column: Chiralpak AD-3 100x4.6 mm i.d., 3 µm; Mobile phase: i-PrOH (0.05% DEA) in _CO2 from 5% to 40%; Flow rate: 3 ml/min; Wavelength: 254 nm.

I44:Rt:=1.918 min.

Intermediate connection I45.

tert-Butyl (3R,8R)-8-fluoro-3,10-dimethyl-11-oxo-1,3,4,7,8,9,10,11-octahydro-2H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2-carboxylate

To a mixture of intermediate I44 (700 mg, 1.70 mmol) in DCM (15 mL) was added DAST (1.09 g, 6.79 mmol) at -40 °C. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was quenched with H2O (20 mL) and _NaHCO3 (sat., aq., 20 mL). The layers were separated and the aqueous phase was extracted with EtOAc (3 x 80 mL). _The combined organic extracts were washed with brine (40 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was combined with another batch (700 mg) and purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 3:1 to 0:1) to give intermediate I45 (470 mg, 93% purity) as a light yellow solid.

Intermediate connection I46.

(3R,8R)-8-Fluoro-3,10-dimethyl-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[ 1,5a][1,4]diazepin-11-one-TFA

Intermediate I46 was prepared according to the procedure described for the synthesis of intermediate I49.

Connection 44.

(3R,8R)-8-Fluoro-3,10-dimethyl-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrαzolo[1,5a][1,4]diazepin- 11-one

Compound 44 was prepared according to the procedure described for the synthesis of compound 46.

MS (ESI): m/z calcd for C ₁₉ H ₁₉ Cl ₂ FN ₄ O ₂ 424; m/z found, 425 [M+H]+; 1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.75-7.66 (m, 2H), 7.43 (dd, J=1.9, 8.2 Hz, 1H), 4.64-4.17 (m, 8H), 3.07 (br s, 3H), 2.95 (br dd, J=5.9, 15.9 Hz, 1H), 2.58 (br d, J=15.8 Hz, 1H), 1.17 (d, J=6.8 Hz, 3H).

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3.2.2.13. Synthesis of compounds 45 and 46

Intermediate compounds I47 and I48.

tert-Butyl (4*S)-4-(hydroxymethyl)-2-methyl-1-oxo-1,4,5,8,9,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d] [ 1,2,5]oxadiazepine-10(2H)-carboxylate tert-Butyl (4*R)-4-(hydroxymethyl)-2-methyl-1-oxo-1,4,5,8,9,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d] [ 1,2,5]oxadiazepine-10(2H)-carboxylate

A mixture of tert-butyl 3-(hydroxy(methyl)carbamoyl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)carboxylate (8.00 g, 27.0 mmol), 3-bromoxetane (4.07 g, 29.7 mmol), TBAI (997 mg, 2.70 mmol) and _Cs2CO3 (13.2 g, 40.5 mmol) in DMF (80 mL) was degassed and purged with N2 ( ₃ times). The reaction mixture was stirred at 70 °C for 3 h under N2 and the mixture was poured into water (200 mL). The mixture was stirred for 5 min and the aqueous phase was extracted with EtOAc (3 x 100 mL). The combined organic extracts were washed with brine (2 x 300 mL), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 100:1 to 1:2) to give a mixture of diastereoisomers (4.50 g, 46%) as a yellow solid.

Diastereoisomers were purified by SFC (column: AD (250 mm x 30 mm, 10 μm); mobile phase: _CO2 /MeOH (with 0.1% _NH3 in H2O), isocratic elution:75:25) to afford intermediate I47 and intermediate I48.

SFC analysis: Column: Chiralpak AD-3 100x4.6 mm i.d., 3 µm; Mobile phase: MeOH (0.05% DEA) in _CO2 from 5% to 40%; Flow rate: 3 ml/min; Wavelength: 220 nm.

I47: Rt:=1.77 min.

I48: Rt:=2.03 min.

Intermediate connection I49.

(4*S)-4-(Hydroxymethyl)-2-methyl-4,5,8,9,10,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5]oxadiazepin-1(2H)-one -TFA

To a solution of intermediate I47 (100 mg, 284 μmol) in DCM (10 mL) was added TFA (1.07 g, 9.36 mmol). The reaction mixture was stirred at 30 °C for 30 min and concentrated

- 85 047929 under reduced pressure to give intermediate compound I49, which was used as such in the next step.

Connection 46.

(4*S)-10-(3,4-Dichlorobenzoyl)-4-(hydroxymethyl)-2-methyl-4,5,8,9,10,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5]oxadiazepin-1(2H)-one

To a mixture of intermediate I49 and _Et3N (91.2 mg, 901 μmol) in DCM (40 mL) was added dropwise a solution of 3,4-dichlorobenzoyl chloride (62.9 mg, 0.30 mmol) in DCM (5 mL) at -10 °C. The reaction mixture was stirred at -10 °C for 2 h and concentrated under reduced pressure. The crude mixture was purified by reversed-phase HPLC (Gilson GX-281 semi-preparative HPLC with Phenomenex Synergi C-18 (10 μm, 150x25 mm) or Boston Green ODS C18 (5 μm, 150x30 mm) and a mobile phase of 5-99% MeCN in H2O (with 0.225% FA) for 10 min and then held at 100% MeCN for 2 min, at a flow rate of 25 mL/min) to afford compound 46 (76 mg, 62%) as a colorless oil.

MS (ESI): m/z calcd for C ₁₈ H ₁₈ Cl ₂ N ₄ O _4, 424; m/z found, 425 [M+H]+; ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.59-7.52 (m, 2H), 7.32-7.30 (m, 1H), 4.99-4.53 (m, 4H), 4.41 (d, J=9.6 Hz, 1H), 4.08-3.74 (m, 4H), 3.29 (s, 3H), 2.92-2.84 (m, 2H), 2.04 (d, J=5.2 Hz, 1H).

Intermediate connection I50.

(4*R)-4-(Hydroxymethyl)-2-methyl-4,5,8,9,10,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5]oxadiazepine-1(2H)-oh-TFA

Intermediate I50 was synthesized according to the procedure described for the synthesis of intermediate I49.

Connection 45.

(4*R)-10-(3,4-Dichlorobenzoyl)-4-(hydroxymethyl)-2-methyl-4,5,8,9,10,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5]oxadiazepin-1(2H)-one

Compound 45 was synthesized according to the procedure described for the synthesis of compound 46.

MS (ESI): m/z calcd for C ₁₈ H ₁₈ Cl ₂ N ₄ O ₄ 424; m/z found, 425 [M+H]+; 1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.57 (s, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.29-7.27 (m, 1H), 4.95-4.66 (m, 2H), 4.56-4.51 (m, 2H), 4.42 -4.40 (m, 1 H), 3.86-3.81 (m, 1H), 3.75-3.72 (m, 3H), 3.26 (s,H) 2.89-2.01 (m, 2H), 2.00 (s, 1H).

- 86 047929

3.2.2.14. Synthesis of compound 47

1) o(^>—Вг Me

HNN J. _TBAI , _Cs2CO3

ALCH t'ME 7ЛСЛ ...

(in; ) ⁰ 2) Split cn using SFC

151

Boc Boc

F

Me

Intermediates I51 and I52.

tert-Butyl (4*S,9R)-4-(hydroxymethyl)-2,9-dimethyl-1-oxo-1,4,5,8,9,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5] oxadiazepine-10(2H)-carboxylate tert-Butyl (4*R,9R)-4-(hydroxymethyl)-2,9-dimethyl-1-oxo-1,4,5,8,9,11 -hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5] oxadiazepine-10(2H)-carboxylate

To a solution of tert-butyl ((3-c)-tert-butyl 3-(hydroxy(methyl)carbamoyl)-6-methyl-6,7-dihydro-N-pyrazolo[4,3- _c ]pyridine-5(4H)-carboxylate (2.30 g, 7.41 mmol) in DMF (20 mL) was added _Cs2CO3 (7.24 g, 22.2 mmol), then 3-bromoxetane (1.32 g, 9.63 mmol) and TBAI (274 mg, 741 μmol). The reaction mixture was stirred at 75 °C for 4 h. The mixture was extracted with EtOAc (2x20 mL) and H2O (20 mL). The organic layer was washed with H2O (2x20 mL), dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient 1: ₁ to 0:1) to give a mixture of diastereoisomers (1.80 g, 56%, 85% purity) as a white solid.

Diastereoisomers were purified by SFC (column: DAICEL CHIRALCEL OD (250 mm x 30 mm, 10 μm); mobile phase: _CO2 /i-PrOH (with 0.1% _NH3 in _H2O ), isocratic elution: 75:25) to afford intermediate I51 and intermediate I52.

SFC analysis: Column: Chiralcel OD-3 100x4.6 mm i.d., 3 µm; Mobile phase: i-PrOH (0.05% DEA) in _CO2 from 5% to 40%; Flow rate: 3 ml/min; Wavelength: 220 nm.

I51: R _t :=1.861 min.

I52: R _t :=2.036 min.

Intermediate connection I53.

tert-Butyl (4*R,9R)-4-(fluoromethyl)-2,9-dimethyl-1-oxo-1,4,5,8,9,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1-d][1,2,5]oxadiazepine-10(2H)-carboxylate

- 87 047929

To a mixture of intermediate I52 (4.32 g, 11.6 mmol) in THF (80 mL) was added 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonyl fluoride (10.5 g, 34.9 mmol) and TREAT-HF (5.63 g, 34.9 mmol), followed by DIPEA (13.5 g, 1045 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 6 h. The reaction mixture was quenched by addition of HCl (1 N, aq, 100 mL) and diluted with water (100 mL). The layers were separated and the aqueous phase was extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, petroleum ether/EtOAc, gradient from 100:1 to 5:1) to give intermediate compound I53 (2.5 g, 58%) as a colorless oil.

Intermediate connection I54.

(4*K,9K)-4-(Fluoromethyl)-2,9-dimethyl-4,5,8,9,10,11-hexahydropyrido[4',3':3,4]pyrazolo[5,1d] [1,2,5]oxadiazepine-1(2H)-oh-TFA ^-F

Intermediate I54 was prepared according to the procedure described for the synthesis of intermediate I49.

Connection 47.

(4*K,9K)-10-(3,4-Dichlorobenzoyl)-4-(fluoromethyl)-2,9-dimethyl-4,5,8,9,10,11-hexahydropyrido[4',3': 3,4]pyrazolo[5,1-d][1 D^oxadiazepin-^SH-one

Compound 47 was synthesized according to the procedure described for the synthesis of compound 46.

MS (ESI): m/z calcd for C ₁₉ H ₁₉ Cl ₂ FN ₄ O ₃ , 440; m/z found, 441 [M+H]+; 1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.54-7.50 (m, 2H), 7.25 (d, J=2.0 Hz, 1H), 5.60-5.44 (m, 2H), 4.71-4.58 (m, 5H), 4.40-4.36 (m, 2H), 3.29 (s, 3H), 3.03 (s, 3H), 2.67 (d, J=15.6 Hz, 1H), 1.26 (s, 3H).

3.2.2.15. Synthesis of compound 48

Intermediate connection I55.

tert-Butyl 11-thioxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine2(7H)-carboxylate

- 88 047929

To a solution of tert-butyl 11-oxo-1,3,4,7,8,9,10,11-octahydro-2H-nirido[4',3':3,4]niαzolo[1,5a][1,4]diazepine-2-carboxylate (250 mg, 816 μmol) in toluene (5 mL) was added Lawesson's reagent (165 mg, 408 μmol). The reaction mixture was stirred at 110 °C for 3 h. The mixture was concentrated under reduced pressure and the residue was purified by preparative TLC (EtOAc) to give intermediate I55 (258 mg, 98%) as a yellow solid.

Intermediate connection I56.

1,2,3,4,7,8,9,10-octahydro-11 H-pyrido[4',3':3,4]pyrazolo[1,5-a] [1,4]diazepine-11-thione hydrochloride

Intermediate I55 (54.0 mg, 0.17 mmol) in i-PrOH (1.07 mL) was charged into a sealed tube. HCl (6 M in i-PrOH, 0.28 mL, 1.68 mmol) was added and the reaction mixture was stirred at 100 °C for 30 min. The mixture was concentrated under reduced pressure. The residue was triturated in DIPE, filtered and dried to give intermediate I56 (35 mg, 81%) as a yellow solid.

Connection 48.

(3,4-Dichlorophenyl)(11-sulfanylidene-1,3,4,7,8,9,10,11-octahydro-2H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepin-2-yl)methanone

Intermediate I56 (35 mg, 135 μmol), 3,4-dichlorobenzoyl chloride (30.7 mg, 0.14 mmol), DCM (0.5 mL), and water (0.5 mL) were charged into a sealed tube. _Na2CO3 (71.7 mg, 0.68 mmol) was added portionwise and the reaction mixture was stirred at room temperature for 1 h. The layers were separated and the organic phase was dried ( _MgSO4 ), filtered, and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (silica, heptane/(EtOAc/EtOH, 3:1), gradient from 100:0 to 0:100). The residue was triturated in DIPE, filtered and dried under vacuum to afford compound 48 (31 mg, 58%) as a pale yellow solid.

¹ H NMR (400 MHz, DMSO-d,6 100°C) δ ppm 10.01-10.29 (m, 1H), 7.66 (d, J=8.1 Hz, 1H), 7.63 (d, J=2.0 Hz, 1H), 7.40 (dd, J=8.1, 2.0 Hz, 1H), 4.70 (s, 2H), 4.26 (t, J=6.9 Hz, 2H), 3.66-3.76 (m, 2H), 3.25 (q, J=6.2 Hz, 2H), 2.71 (t, J=5.9 Hz, 2H), 2.20 (quin, J=6.8 Hz, 2H); LCMS (method C): R _t =1.01 min, m/z calcd. for C ₁₇ H ₁₆ Cl ₂ N ₄ OS 394.0, mass/charge, found value 395.1 [M+H]+.

3.2.2.15. Synthesis of compound C51 and analogues

To a solution of 3,4-difluorobenzoic acid (30 mg, 0.19 mmol) in N,N-dimethylformamide (3 ml) was added 2-(7-azabenzotriazol-1-yl)-N,N,N-tetramethyluronium hexafluorophosphate (105 mg, 0.276 mmol). Then intermediate I90 (50 mg, 0.14 mmol, purity 95%) and N,N-diisopropylethylamine (90 mg, 0.70 mmol) were added. The mixture was stirred at room temperature overnight. The mixture was poured into water (10 ml) and extracted twice with ethyl acetate (10 ml). The combined organic phases were washed with brine (5 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified using

- 89 047929 quantitative HPLC (Column: Waters Xbrige C18 (5 μm 19x150 mm), Mobile Phase A: Water (0.1% Ammonium Bicarbonate), Mobile Phase B: Acetonitrile, UV: 214 nm, Flow Rate: 15 mL/min, Gradient: 20-75% (%B)) to afford compound C51 (35 mg, yield 56%, purity 98.7% according to LCMS) as a white solid.

The compounds listed in the table below were prepared in a similar manner.

Connection No. Structure Reagents Exit C53 O U'M /= ⁰ M3“°\ 1. Intermediate 190 2. 3-Fluoro-4-methylbenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 32% C49 XX/ОУу about A~N ,=-. ⁰ 1. Intermediate 190 2. 4-Fluoro-3-methylbenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 31% C52 _X1 J Χ ^ν % ^Ν I /=\ ⁰ 1. Intermediate 190 2. 3-Cyano-4-fluorobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 90% C54 ХХУл ο A'N =, ⁰ MX ⁰ ,. 1. Intermediate 190 2. 3-Chlorobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 26% C50 o' ί)' U 1. Intermediate 190 2. 4-Chlorobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 39%

- 90 047929

C62 F O ⁰ 1. Intermediate 190 2. 3-Chloro-4- (difluoromethyl)benzoic acid 160 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 43% C57 o [u) IL- 1. Intermediate 190 2. 4-Chloro-3(difluoromethyl)benzoic acid 164 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 29% C56 0 /=\ 1. Intermediate 190 2. 3,4,5-Trifluorobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 62% C64 JUL OjU ^Ν I JL /=\ ⁰ UiL 1. Intermediate 190 2. 4-Chloro-3-cyanobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 59% C70 F II Ί ϊ J” _ ⁰ 1. Intermediate 190 2. 3-Cyano-4- (difluoromethyl)benzoic acid 169 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 35% C68 F ^F Τ 1 ι Τ ' ^ν ~λ D >-Ν .~ ₍ ⁰ ΗΖΗ 1. Intermediate 190 2. 4-(Difluoromethyl)-3-fluorobenzoic acid 171 3. HATU 4. Ν, N-Diisopropylethylamine 5. DMF 33%

3.2.2.16. Synthesis of compound C59 and analogues

37444-46-5 I12 C59

To a solution of pyridazin-3-ylmethanol (220.0 mg, 2.0 mmol) in DCM (10 mL) was added a solution of sulfur dichloride (595.0 mg, 5.0 mmol) in DCM (2 mL) dropwise at 0 °C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated to give crude intermediate I112 (200.0 mg, yield 78.1%, purity 90% by ¹ H NMR) as a yellow oil. ¹ H NMR (400 MHz, CD3OD) δ 9.71-9.69 (m, 1H), 8.77 (s, 1H), 8.64-8.62 (m, 1H), 5.13-5.12 (m, 2H).

To a solution of intermediate I112 (100 mg, 0.26 mmol, the preparation was reported in EP19176933) in DMF (2 mL) was added 60 wt% NaH in mineral oil (16.0 mg, 0.4 mmol) at 0 °C under nitrogen atmosphere. The mixture was stirred at 0 °C for 30 min. Then, intermediate I14 (40.0 mg, 0.31 mmol) was added to this mixture. The mixture was stirred at 25 °C for 3 h. The mixture was treated with water (10 mL) at 0 °C and stirred for 10 min. Then, EtOAc (10 mL) was added three times to the extract of the desired compound. The combined organic phases were washed with brine

- 91 047929 with brine (30 mL), dried over _Na2SO4 , filtered and concentrated. The residue was purified by preparative HPLC to afford compound C59 (41.0 mg, 33.6% yield, 98.2% purity by LCMS) as _a pale yellow solid.

The compounds listed in the table below were prepared in a similar manner._________

Connection No. Structure Reagents Exit C60 C65 su sk su SK O li G <У n-A 1. Intermediate 114 2. 2-(Chloromethyl)pyrimidine 3.NaH 4. DMF 1. Intermediate 114 2. 2-(Bromomethyl)pyrazine 3. NaH 4. DMF 36.0% 23.6% C58 a^, cP N /'U N- o -N i 1. Intermediate 114 2. 1-(2-Bromoethyl)-4- methoxybenzene 3.NaH 4. DMF 19.5% C69 Ск I Cl^ Ί K O _-N W 0 V F /=\ KF 4 y-o 1. Intermediate 114 2. 2-(Bromomethyl)-5(difluoromethoxy)pyridine 3.NaH 4. N,N-Dimethylformamide 18% C66 SK sg \£. _н ' ί \=о γ N- o' n /=\ о F 1. Intermediate 114 2. 1-(Bromomethyl)- 3 (difluoromethoxy)benzene 3.NaH 4. N,N-Dimethylformamide 64% C67 CI C!' Ί O XL' ¹ ^ A—N 0 (/ 1. Intermediate 114 2. 1-(Bromomethyl)2(difluoromethoxy)benzene 3.NaH 4. N,N-Dimethylformamide 64%

3.2.2.17. Synthesis of compound C81 and analogues

A mixture of 1-(chloromethyl)-3-fluorobenzene (70 mg, 0.18 mmol), intermediate I14 (31 mg, 0.21 mmol) and cesium carbonate (176 mg, 0.56 mmol) in N,N-dimethylformamide (2 mL) was stirred at room temperature for 3 h. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL). _The organic layer was washed with brine (10 mL), dried over _Na2SO4 and concentrated. The residue was purified by silica gel chromatography (dichloromethane:methanol=50:1) and further purified by preparative TLC (dichloromethane:methanol=20:1) to give compound C81 (37 mg, yield 42%, purity 97.35% by LCMS) as a white solid.

The compounds listed in the table below were prepared in a similar manner.__________

Connection No. Structure Reagents Exit C82 g ώ and o 1. Intermediate 114 2. 4-(Chloromethyl)benzonitrile 3. Cesium carbonate 4. N,N-Dimethylformamide 37%

- 92 047929

A tube was charged with 2-(3,4-dichlorobenzoyl)-1,2,3,4,7,8,9,10-octahydro-11H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one 33 (73 mg, 0.19 mmol) in dry DMF (1.2 mL) in N ₂ -amM. NaH 60 wt % in mineral oil (38.5 mg, 0.96 mmol) was added and the mixture was stirred for 1 h at rt. 2-(Chloromethyl)pyridine hydrochloride (66 mg, 0.39 mmol) was added and the mixture was stirred at rt for 16 h. The reaction mixture was quenched with water and extracted with Me-THF. The organic layer was separated, dried over _MgSO4 , filtered and concentrated in vacuo. _The residue was purified by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 μm, 30x150 mm, mobile phase: 0.25% _NH4HCO3 in water, _CH3CN ). The residue was dissolved in MeOH and concentrated in vacuo to give 2-(3,4-dichlorobenzoyl)-10-(pyridin-2-ylmethyl)-1,2,3,4,7,8,9,10-octahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11-one C83 (18.8 mg, 21% yield) as a white foam.

3.2.2.19. Synthesis of compound C63

To a solution of 4-methoxypyridine (10.5 g, 96.12 mmol) in tetrahydrofuran (100 mL) was added di-tert-butyl dicarbonate (21.0 g, 96.22 mmol) portionwise at 0 °C under nitrogen atmosphere. Then, 1 M vinyl magnesium bromide in tetrahydrofuran (100 mL, 100 mmol) was added dropwise to the mixture over 30 min at 0 °C. The mixture was stirred at room temperature for 3 h. The mixture was treated with hydrochloride (1 M, 200 mL) at 5 °C and stirred for 10 min. Then, the mixture was extracted three times with ethyl acetate (100 mL). The combined organic layers were washed with 10% aqueous sodium bicarbonate solution (100 mL), then brine (100 mL), dried over Na ₂ SO ₄ and filtered. The filtrate was evaporated to give intermediate compound I113 (8.5 g, yield 35%, purity 90% by ¹ H NMR). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.79 (s, 1H), 5.84-5.76 (m, 1H), 5.30-5.11 (m, 3H), 5.09-5.02 (m, 1H), 2.93-2.87 (m, 1H), 2.54-2.51 (m, 1H), 1.47 (s, 9H).

To a suspension of intermediate compound I113 (9.00 g, 36.28 mmol, 90% purity) in acetic acid (50 mL) was added activated zinc dust (12.1 g, 185.0 mmol) at 25°C. After the addition

- 93 047929 The resulting mixture was stirred at 70°C for 14 h. The mixture was filtered and the filter cake was washed with ethanol (200 mL). The filtrate was concentrated in vacuo to dryness. The residue was purified on a C18 column (mobile phase A: water (+ 0.1% ammonium bicarbonate), mobile phase B: acetonitrile, gradient: 05-55% (%B)) to give intermediate compound I114 (2.31 g, yield 25%, purity 90% according to ¹ H NMR) as a brown oil. ^1H NMR (300 MHz, CDCl ₃ ) δ 5.855.74 (m, 1H), 5.30-5.15 (m, 3H), 4.23-4.15 (m, 1H), 3.40-3.30 (m, 1H), 2.77-2.33 (m, 4H), 1.53 (s, 9H).

To a solution of intermediate I114 (10.5 g, 41.9 mmol, 90% purity) in tetrahydrofuran (50 mL) was added lithium bis(trimethylsilyl)amide (63 mL, 63 mmol) dropwise at -65 °C. The mixture was stirred at -65 °C for 30 min. Diethyl oxalate (7.8 g, 53.8 mmol) was then added and the mixture was stirred at -65 °C to room temperature overnight. The reaction mixture was poured into water (100 mL) and extracted three times with dichloromethane (50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the crude material. The crude material was purified by silica gel chromatography (petroleum ether:ethyl acetate = 3:1) to afford intermediate I115 (7.5 g) as a yellow oil. LCMS (ESI): Rt = 2.210 min, mass, calcd. For C ₁₆ H ₂₃ NO ₆ 325.15, mass/charge, found value 270.1 [M+H-56]+.

A solution of intermediate I115 (7.5 g, 16.13 mmol, 70% purity) and hydrazine hydrate (1.5 g, 39.8 mmol, 85% purity) in glacial acetic acid (20 ml) was stirred at 100 °C for 4 h. The reaction mixture was poured into water (100 ml) and extracted three times with dichloromethane (200 ml). The combined organic layers were washed with brine (100 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give a residue which was purified by silica gel chromatography (petroleum ether:ethyl acetate=1:1) to give intermediate I116 (1.2 g, yield 27%, purity 90% by LCMS) as a yellow solid. 1H NMR (400 MHz, CDCl ₃ ) δ 5.75-5.66 (m, 1H), 5.29-5.49 (m, 4H), 4.43-4.33 (m, 2H), 4.19-4.13 (m, 1H), 3.05-2.92 (m, 2H), 1.47 (s, 9H), 1.3 (t, J=7.2 Hz, 3H).

To a mixture of intermediate I116 (570 mg, 1.60 mmol, 90% purity) in tetrahydrofuran (30 mL) at 0 °C were added sequentially benzyl (2-hydroxyethyl)carbamate (360 mg, 1.84 mmol), triphenylphosphine (670 mg, 2.55 mmol) and diisopropyl azodicarboxylate (610 mg, 3.02 mmol). After addition, the mixture was stirred at 0 °C to room temperature overnight. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (mobile phase A: water (+0.1% ammonium bicarbonate), mobile phase B: acetonitrile, gradient: 05-65% (%B)) to give intermediate compound I117 (437 mg, yield 55%, purity 100% by LCMS) as a yellow oil. LCMS (ESI): R _t = 1.78 min, mass, calcd. For CC ₂₆ H ₃₄ N ₄ O ₆ 498.25, mass/charge, found value 499.1 [M+H]+.

To a mixture of intermediate I117 (230 mg, 0.46 mmol, 100% purity) in ethanol (15 mL) was added 10% palladium on activated carbon (ca. 50% wetted, 23 mg) at room temperature. The mixture was stirred at room temperature under hydrogen atmosphere (1 atm) for 16 h. The mixture was filtered. The filtrate was concentrated in vacuo to give intermediate I118 (200 mg, 81% yield, 68% purity by LCMS) as a gray oil. LCMS (ESI): R _t = 1.62 min, mass calcd. For C ₁₈ H ₃₀ N ₄ O ₄ 366.23, m/z, found value 367.1 [M+H]+.

To a mixture of intermediate I118 (200 mg, 0.37 mmol, 68% purity) in 1,4-dioxane (8 mL) was added 2,3,4,6,7,8-hexahydro-1H-pyrimido[1,2-a]pyrimidine (20 mg, 0.14 mmol) at room temperature. The mixture was stirred at 100 °C for 2 h. The mixture was concentrated in vacuo to give a residue, which was purified by a C18 column (mobile phase A: water (+0.1% ammonium bicarbonate), mobile phase B: acetonitrile, gradient: 05-45% (%B)) to give intermediate I119 (95 mg, 80% yield, 100% purity by LCMS) as a white solid. LCMS (ESI): R _t = 1.45 min, mass, calc. For C ₁₆ H ₂₄ N ₄ O ₃ 320.18, mass/charge, found value 321.1 [M+H]+.

To an ice-cooled mixture of 60 wt % sodium hydride in mineral oil (24 mg, 0.60 mmol) in NH-dimethylformamide (4 mL) was added intermediate compound I119 (125 mg, 0.39 mmol, 100% pure) at a temperature below 10 °C. After stirring the mixture at 10 °C for 30 min, a solution of 4-methoxybenzyl chloride (73 mg, 0.47 mmol) in N,N-dimethylformamide (1 mL) was added dropwise. The mixture was stirred at 10 °C for 120 min. The mixture was then quenched with water (50 mL) and extracted twice with ethyl ether (30 mL). The combined organic layers were washed with brine (80 mL), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo to give intermediate I120 (198 mg, 100% yield, 87% purity by LCMS) as a grey oil. LCMS (ESI): R _t = 1.71 min, mass, calcd. For C ₂₄ H ₃₂ N ₄ O ₄ 440.24, mass/charge, found 441.1 [M+H]+.

To a solution of intermediate I120 (198 mg, 0.39 mmol, 87% purity) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The resulting mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to dryness. The residue was then diluted with dichloromethane (50 mL) and neutralized with saturated aqueous

- 94 047929 sodium bicarbonate solution (50 mL). The mixture was extracted twice with dichloromethane (50 mL). The combined organic layers were washed with brine (50 mL) and dried over sodium sulfate. The mixture was concentrated in vacuo to give intermediate I121 ( ₁₄₆ mg, 99% yield, 90% purity by LCMS) as a gray solid. LCMS (ESI): Rt=1.36 min, mass, calcd. For _{C19H24N4O2 340.19} , m/z, found 341.1 [M ₊ H]+.

To a mixture of intermediate I121 (145 mg, 0.38 mmol, 100% purity) and 3,4-dichlorobenzoic acid (100 mg, 0.52 mmol) in dichloromethane (15 ml) were added X,X-diisopropylethylamine (250 mg, 1.93 mmol) and O-(7-α-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (290 mg, 0.76 mmol) at room temperature.

The resulting mixture was stirred at 25°C for 16 h. The mixture was concentrated in vacuo to give a residue, which was purified using a C18 column (mobile phase A: water (+ 0.02% ammonium acetate), mobile phase B: acetonitrile, gradient: 05-60% (%B)) to give compound C63 (79 mg, yield 40%, purity 99% by LCMS) as a white solid.

3.2.2.20. Synthesis of the compound C61*R _Bg '-'. ^on EUN

2, Boc ₂ O, TEA, DCM

C61‘R

1128

To a solution of (4-methoxyphenyl)methanamine (16.3 g, 98% pure, 116 mmol) in ethanol (60 mL) was added 2-bromoethanol (5.00 g, 96% pure, 38.4 mmol). After stirring at 85 °C overnight, the mixture was concentrated in vacuo to dryness. The residue was dissolved in dichloromethane (60 mL) and triethylamine (7.80 g, 77.083 mmol), di-tert-butyl dicarbonate (33.6 g, 154 mmol) were added. The mixture was stirred at room temperature for 2 h. The mixture was treated with saturated aqueous ammonium chloride solution (100 mL), brine (50 mL) and extracted three times with dichloromethane (50 mL). The combined organic phase was dried over _Na2SO4 ( _s ), filtered and concentrated to give the crude product which was purified by column chromatography on silica gel (petroleum ether:ethyl acetate=10:1 to 2:1) to give intermediate I122 (6.88 g, 90% purity by ^1H NMR, 57% yield) as a yellow oil. LCMS (ESI): Rt=1.36 min, mass calcd for C15H23NO4 281.16, mass/charge found 226.1 [M-56+H] ⁺ . 1H NMR (400 MHz, CDCl3) 7.16 (d, J=6.8 Hz, 2H), 6.86 (d, J=8.4 Hz, 2H), 4.41 (s, 2H), 3.80 (s, 3H), 3.67 (s, 2H), 3.37 (s, 2H), 3.12 (br s, 1H), 1.48 (s, 9H).

To a solution of intermediate I92*R (904 mg, 2.49 mmol) in ethyl acetate (4 mL) was added 3 M HCl/EA (20 mL, 60 mmol) at room temperature. After stirring at room temperature for 4 h, the mixture was concentrated in vacuo to give intermediate I123 (715 mg, 90% purity by 1H NMR, 86% yield) as a yellow solid, which was used in the next step without further purification. 1H NMR (400 MHz, DMSO-d ₆ ) 10.95 (br s, 3H),

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4.77-4.70 (m, 1H), 4.42-4.28 (m, 4H), 3.31-3.26 (m, 1H), 3.11-3.04 (m, 1H ), 1.31 (t, J=7.2 Hz, 3H). LCMS (ESI): R _t =1.50 min, mass, calc. for C ₁₀ H ₁₃ ClF ₃ N ₃ O ₂ 299.1, mass/charge, found value 264.2 [MHC1+H]+.

To a solution of intermediate I123 (120 mg, 90% purity, 0.360 mmol) in pyridine (5 mL) was added 3,4-dichlorobenzoyl chloride (400 mg, 1.91 mmol) in dichloromethane (1 mL) at 0 °C. The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 80%) to give intermediate I124 (340 mg, 55% purity, 83% yield) as a yellow solid. LCMS (ESI): R _t = 2.056 min, mass calcd. For C ₂₄ H ₁₆ Cl ₄ F ₃ N ₃ O ₄ 609.21, mass/charge, the found value is 610.0 [M+H] ⁺ .

A mixture of intermediate I124 (340 mg, 55% purity, 0.307 mmol) and potassium carbonate (120 mg, 0.868 mmol) in ethanol (10 mL) was stirred at 40 °C for 2 h. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water (0.1% ammonium bicarbonate = 10% to 70%) to give intermediate I125 (120 mg, 90% purity by NMR, 81% yield) as a white solid. LCMS (ESI): R _t = 1.67 min, mass calcd. For C17H ₁₄ Cl ₂ F ₃ N ₃ O ₃ 435.04, mass/charge, found 435.9 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) 7.657.51 (m, 2H), 7.40-7.30 (m, 1H), 5.95 (br s, 0.7H), 5.58 (br s, 0.3H), 4.91-4.22 (m, 5H), 3.33-3.15 (m, 2H), 1.26 (t, J=7.2 Hz, 3H).

To a mixture of intermediate I125 (120 mg, 90% purity, 0.248 mmol), intermediate I122 (150 mg, 90% purity, 0.480 mmol) and triphenylphosphine (130 mg, 0.496 mmol) in tetrahydrofuran (5 mL) was added diisopropyl azodicarboxylate (100 mg, 0.495 mmol) at 0 °C. The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 80%) to give intermediate I126 (30 mg, 100% purity, 17% yield) as a yellow solid. LCMS (ESI): R _t = 2.047 min, mass, calc. For C ₃₂ H ₃₅ Cl ₂ F ₃ N ₄ O ₆ 698.19, mass/charge, found value 699.2 [M+H] ⁺ .

A mixture of intermediate I126 (35 mg, 0.05 mmol) and lithium hydroxide hydrate (11 mg, 0.25 mmol) in tetrahydrofuran/water (0.5 ml/0.2 ml) was stirred at 25 °C for 5 h. The mixture was concentrated to give crude intermediate I127 (50 mg, crude, 60% purity by LCMS) as a white solid, which was used directly in the next step without further purification. LCMS (ESI): R _t = 1.49 min, mass, calcd. For C ₃₀ H ₃₁ Cl ₂ F ₃ N ₄ O ₆ 670.16, m/z, found 671.5 [M+H] ⁺ .

A solution of intermediate I127 (50 mg, 0.0744 mmol) in 4 M HCl/EtOAc (2 mL) was stirred at 25 °C for 3 h. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 80%) to give intermediate I128 (29 mg, 89% yield, 40% purity by LCMS) as a white solid. LCMS (ESI): R _t = 1.42 min, mass calcd. For C ₂₅ H ₂₃ Cl ₂ F ₃ N ₄ O ₄ 570.1, mass/charge, found 571.4 [M+H] ⁺ .

A mixture of intermediate I128 (25 mg, 0.044 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (25 mg, 0.066 mmol) and N,N-diisopropylethylamine (17 mg, 0.132 mmol) in X,X-dimethylformamide (1 mL) was stirred at room temperature for 1 h. The mixture was purified using a C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 80%) to give compound C61*R (9 mg, 38% yield, 99.67% purity by LCMS) as a white solid.

3.2.2.21. Synthesis of compound C61*S РМВ ,....... ПН ³ 'Ср. __ Вос-' DIAD. PPh ₃₁ THF > 0 <I92'S 112 FjC, n ^F = ^C ' _r ^НС|/ЕА _г HATU. DIEA PMB DMF HO 1131 ¹¹³ η 1 I'S Г n—, Ci ^ n'· \ > 0 .% ^N , , ⁰ / C6VS FjC' J—\ Boc LiOH G '3 p . Boe 4 ” Boc'' —N = ₀ PMB MeOH/HjO PMB HO ' iso 9 01,,¾. Cr^Y ^CI Ο ΓΜΗ ^TEA °™ 2

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To a mixture of intermediate I92*S (0.8 g, 2.20 mmol), intermediate I122 (1.0 g, 90% pure, 3.20 mmol) and triphenylphosphine (1.1 g, 4.19 mmol) in tetrahydrofuran (15 mL) was added diisopropyl azodicarboxylate (0.85 g, 4.20 mmol) at 0 °C. The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 80%) to give intermediate I129 (1.0 g, 90% pure by NMR, 65% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) 7.12-7.04 (m, 2H), 6.82 (d, J=8.4 Hz, 2H), 5.42-5.31 (m, 0.5H), 5.16-5.03 (m, 1.5H), 4.79-4.58 (m, 2H), 4.42-4.05 (m, 5H), 3.78 (s, 3H), 3.65-3.42 (m, 2H), 3.14-3.02 (m, 2H), 1.52 (s, 9H), 1.43-1.38 (m, 12H).

To a solution of intermediate I129 (550 mg, 90% purity, 0.790 mmol) in methanol (30 mL) was added lithium hydroxide monohydrate (300 mg, 7.15 mmol) in 20 mL of water. The reaction mixture was then stirred at 25 °C for 3 h. The mixture was concentrated in vacuo to remove the organic solvent. The aqueous layer was diluted with water (100 mL) and acidified with 1 M hydrochloric acid to pH ~5. The mixture was extracted twice with ethyl acetate (100 mL). _The combined organic layers were dried over _Na2SO4 (solid), filtered and concentrated to give intermediate I130 (450 mg, 95% purity by NMR, 90% yield) as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) 7.15-7.02 (m, 2H), 6.85-6.78 (m, 2H), 5.39 (br s, 0.5H), 5.19-5.05 (m, 1.5H), 4.78-4.61 (m, 2H), 4.31-4.12 (m, 3H), 3.77 (s, 3H), 3.68-3.46 (m, 2H), 3.18-3.03 (m, 2H), 1.54-1.50 (m, 9H), 1.41 (s, 9H).

A solution of intermediate I130 (450 mg, 95% purity, 0.714 mmol) in HCl/ethyl acetate (30 mL) was stirred at 25 °C for 3 h. The mixture was concentrated in vacuo to give intermediate I131 (240 mg, 100% purity, 72% yield) as a yellow solid. LCMS (ESI): Rt=1.142 min, mass calcd for C ₁₈ H ₂₁ F ₃ N ₄ O ₃ 398.16, mass/charge found 399.2 [M+H]+.

A mixture of intermediate I131 (140 mg, 100% purity, 0.301 mmol), 2-(7-azabenzotriazol-1-yl)-N,N-tetramethyluronium hexafluorophosphate (160 mg, 0.421 mmol) and N,N-diisopropylethylamine (200 mg, 1.55 mmol) in N-dimethylformamide (15 mL) was stirred at room temperature for 2 h. The reaction mixture was treated with ice water (50 mL) and extracted twice with ethyl acetate (50 mL). The combined organic phases were washed twice with brine (50 mL). _The organic layer was dried over _Na2SO4 (solid), filtered and concentrated. The residue was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 5% to 40%) to give intermediate compound I132 (100 mg, purity 97%, yield 85%) as a yellow solid. 1H NMR (400 MHz, MeOD) 7.26 (d, J=8.4 Hz, 2H), 6.90 (d, J=8.0 Hz, 2H), 4.65 (s, 2H), 4.23-4.28 (m, 3H), 3.98 (d, J=16.8 Hz, 1H), 3.77 (s, 3H), 3.69 (t, J=6.4 Hz, 2H), 3.65-3.55 (m, 1H), 2.97-2.92 (m, 1H), 2.77-2.70 (m, 1H).

To a solution of intermediate I132 (100 mg, 97% purity, 0.255 mmol) in pyridine (5 mL) was added 3,4-dichlorobenzoyl chloride (100 mg, 0.477 mmol) in dichloromethane (1 mL) at 0 °C. The mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure. The residue was purified by C18 column (acetonitrile:water containing 0.1% ammonium bicarbonate = 10% to 70%) to give compound C61*S (60 mg, purity 99.5%, yield 42%) as a white solid.

3.2.2.22. Synthesis of C55*R and C55*S compounds

A mixture of intermediate I99 (6 g, 17.4 mmol) in 4 M HCl/EtOAc (50 mL) was stirred at room temperature for 1 h. The mixture was then concentrated in vacuo to dryness. The residue was neutralized with saturated aqueous sodium hydrogen carbonate (50 mL) and extracted three times with EtOAc (50 mL). The combined organic phases were washed with brine (50 mL),

- 97 047929 was dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated under reduced pressure to give intermediate compound I133 (4.5 g, 90% purity by 1H NMR, 95% yield) as a yellow solid. 1H NMR (400 MHz, CDCl ₃ ) δ 6.01-5.69 (m, 1H), 4.33-4.20 (m, 3H), 3.96-3.75 (m, 1H), 3.23-3.13 (m, 1H), 2.88-2.83 (m, 1H), 2.69-2.62 (m, 1H), 2.47-2.32 (m, 3H), 2.07-2.04 (m, 1H), 1.31 (t, J=7.2 Hz, 3H).

To a solution of 3,4-dichlorobenzoic acid (3.5 g, 18 mmol), 2-(7-azabenzotriazol-1-yl)-H,N,N',N'-tetramethyluronium hexafluorophosphate (6.8 g, 18 mmol), and NN-diisopropylethylamine (4.6 g, 3.6 mmol) in N,N-dimethylformamide (25 mL) was added intermediate I133 (3.1 g, 95% purity, 12 mmol). The mixture was then stirred at room temperature overnight. The mixture was diluted with ethyl acetate (30 mL) and washed with brine (20 mL), dried over _Na2SO4 (solid), and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 4:1) to give intermediate compound I134 (3 g, purity 90%, yield 56%) as a yellow solid. 1H NMR (400 MHz, _CDCl3 ) δ 7.55-7.56 (m, 2H), 7.32 (s, 1H), 6.03-5.41 (m, 2H), 4.82-4.07 (m, 4H), 3.15 (br s, 2H), 1.26 (t, J=7.2 Hz, 3H).

To a solution of intermediate I134 (3 g, 90% purity, 6.48 mmol) and tert-butyl (2-bromoethyl)carbamate (2.88 g, 18 mmol) in acetonitrile (50 mL) was added cesium carbonate (6.3 g, 19.4 mmol) at room temperature. The mixture was then stirred at room temperature overnight. The mixture was concentrated under reduced pressure to dryness, the residue was diluted with ethyl acetate (50 mL) and washed twice with water (50 mL) and brine (50 mL). The organic phase was dried over _Na2SO4 (solid), filtered and concentrated to give a residue which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1 to 1:1) to give intermediate compound I135 (1.9 g, purity 90%, yield 43%) as a yellow oil. 1H NMR (400 MHz, _CDCl3 ) δ 7.55-7.45 (m, 2H), 3.48 (s, 2H), 3.09-3.03 (m, 2H), 1.40 (s, 12H).

A mixture of intermediate I135 (2.3 g, 3.7 mmol) in 4 M HCl/EtOAc (30 mL) was stirred at room temperature for 1 h. The residue was neutralized with saturated aqueous sodium hydrogen carbonate (30 mL) and extracted three times with ethyl acetate (30 mL). _The combined organic phases were washed with brine (20 mL), dried over _Na2SO4 and filtered. The filtrate was concentrated in vacuo to give intermediate I136 (1.5 g, 90% purity, 91% yield) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 7.79-7.73 (m, 2H), 7.47-7.46 (m, 1H), 6.41-6.01 (m, 1H), 5.43-5.32 (m, 1H), 4.83-4.11 (m, 8H), 3.01 (s, 3H), 1.36-1.06 (m, 3H).

To a solution of intermediate I136 (600 mg, 90% purity, 1.2 mmol) in 1,4-dioxane (15 mL) was added 2,3,4,6,7,8-hexahydro-1H-pyrimido[1,2-a]pyrimidine (51 mg, 0.36 mmol) at room temperature. The reaction mixture was stirred at 100 °C under nitrogen for 2 h. The mixture was concentrated under reduced pressure to dryness. The residue was dissolved in ethyl acetate (25 mL) and the mixture was washed twice with water (15 mL) and brine (15 mL). The organic phase was dried over _Na2SO4 (solid), filtered and concentrated to give intermediate I137 (500 mg, 90% purity, 93% yield) as a white solid. 1H NMR (400 MHz, DMSO- _d6 ) δ 8.25-8.19 (m, 1H), 7.75 (s, 2H), 7.47-7.46 (m, 1H), 6.28-5.99 (m, 1H), 5.40-5.26 (m, 1H), 4.60-4.02 (m, 4H), 3.60-3.57 (m, 2H), 3.27-3.18 (m, 4H).

To a solution of intermediate I137 (400 mg, 90% purity, 0.88 mmol) in N,N-dimethylformamide (8 mL) were added 1-(chloromethyl)-4-methoxybenzene (320 mg, 1 mmol) and sodium hydride (80 mg, 60 wt%, 1.76 mmol) at room temperature. The mixture was then stirred at room temperature for 2 h. The mixture was quenched with aq. _NH4Cl (25 mL) and extracted three times with EtOAc (25 mL), the combined organic phases were washed twice with water (15 mL) and brine (15 mL), dried over _Na2SO4 (solid), and filtered. The filtrate was concentrated in vacuo to afford compound C55 (300 mg, 90% purity, 60% yield) as a white solid. 1H NMR (400 MHz, CDC13) δ 7.56-7.53 (m, 2H), 7.32-7.30 (m, 1H), 7.26-7.21 (m, 2H), 6.91-6.83 (m, 2H), 6.03-5.48 (m, 2H), 4.70-4.62 (m, 2H), 4.26 (s, 2H), 3.86-3.78 (m, 3H), 3.62 (s, 2H), 3.11-3.89 (m, 2H), 1.58 (s, 2H), 0.88-0.83 (m, 1H).

The racemic mixture of compound C55 (300 mg, 90% purity, 0.56 mmol) was separated by chiral preparative HPLC (separation condition: column: Chiralpak IB 4.6 μm, 20*250 mm; mobile phase: hexane: EtOH: DEA=95:5:0.2 at 15 mL/min; temperature: 30°C; wavelength: 230 nm) to afford compounds C55*R (56.3 mg, 99% purity, 38% yield, 99.9% ee) and C55*S (59 mg, 99% purity, 38% yield, 99.9% ee) as yellow solids.

Compound C55*R: chiral HPLC (Chiralpak IB 5 μm, 4.6 * 250 mm; mobile phase: IE-hexaneEtOH-40-60. LSM at 1 ml/min; temperature: 30°C; wavelength: 254 nm, R _T = 11.173 min).

Compound C55*S: chiral HPLC (Chiralpak IB 5 μm, 4.6 * 250 mm; mobile phase: IE-hexaneEtOH-40-60. LSM at 1 ml/min; temperature: 30°C; wavelength: 254 nm, R _T = 12.795 min).

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3.2.2.23. Synthesis of compound C79

A mixture of the intermediate, 5-(tert-butyl)-3-ethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (1 g, 3.39 mmol), and lithium hydroxide hydrate (427 mg, 0.48 mmol) in tetrahydrofuran/water (10 mL/3 mL) was stirred at room temperature overnight. The mixture was diluted with water (10 mL) and acidified with 1N hydrochloric acid to pH = 6. The mixture was then extracted three times with ethyl acetate (10 mL). The combined organic layers were washed with brine (20 mL), dried over _Na2SO4 and concentrated in vacuo to give intermediate I138 (200 mg, 22% yield, 100% purity by LCMS) as yellow solids. _{LCMS (ESI): Rt=0.94 min, mass, calcd. For C12H17N3O4 267.1 , m} /z, found 266.2 [MH] _- ^.

1H NMR (400 MHz, DMSO-d6) δ 4.48 (s, 2H), 3.60-3.57 (m, 2H), 2.67-2.64 (m, 2H), 1.41 (s, 9H).

To a mixture of pyridin-2-ylmethanamine (1.12 g, 10.4 mmol), N-diisopropylethylamine (4.0 g, 31.08 mmol) in tetrahydrofuran (20 mL) was added ethyl 2-(bromomethyl)acrylate (2.00 g, 10.4 mmol). The mixture was then stirred at room temperature overnight. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (100 mL). The organic layer was concentrated in vacuo to give a residue, which was purified by column chromatography on silica gel (dichloromethane:methanol = 50:1) to give intermediate compound I139 (1.0 g, yield 43%, purity 80% by LCMS) as a yellow oil. LCMS (ESI): Rt = 1.24 min, mass calcd. For C ₁₂ H ₁₆ N ₂ O ₂ 220.1, mass/charge, the found value is 221.1 [M+H]+.

A mixture of intermediate I138 (1.21 g, 4.54 mmol), intermediate I139 (1.00 g, 4.54 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (2.59 g, 6.81 mmol) and N-diisopropylethylamine (1.76 g, 13.6 mmol) in H2-dimethylformamide (20 mL) was stirred at room temperature overnight. The reaction mixture was poured into water (50 mL) and extracted three times with ethyl acetate (30 mL). The combined organic layers were washed with brine (30 mL) and concentrated. The residue was purified by column chromatography on silica gel (dichloromethane:methanol=50:1) to afford intermediate compound I140 (300 mg, yield 14%, purity 80% by LCMS) as a yellow oil. LCMS (ESI): Rt=1.50 min, mass, calcd. For _{C24H31N5O5 469.2} , mass/charge, found value 470.5 [M _{+ H} ] ⁺ .

A mixture of intermediate I140 (300 mg, 0.64 mmol) and 1,8-diazabicyclo[5.4.0]-7-undecene (48 mg, 0.32 mmol) in acetonitrile (10 mL) was stirred at 50 °C for 3 h under nitrogen. The mixture was then concentrated in vacuo to give a residue, which was purified by column

- 99 047929 silica gel chromatography (dichloromethane:methanol=50:1) to give intermediate I141 (120 mg, yield 40%, purity 80% by LCMS) as a yellow oil. LCMS (ESI): Rt=1.51 min, mass, calcd. For _{C24H3iN5O5 469.2} , mass/charge, found value 470.5 [M+H] ₊ .

A mixture of intermediate I141 (120 mg, 0.26 mmol) and lithium hydroxide hydrate (32 mg, 0.767 mmol) in tetrahydrofuran/water (2:1, 3 ml) was stirred at room temperature for 5 h. The mixture was then acidified with 1N hydrochloric acid to pH=6. The mixture was concentrated in vacuo to give crude intermediate I142 (200 mg, crude) as a yellow oil, which was used directly in the next step.

A mixture of crude intermediate I142 (200 mg crude, 0.45 mmol), methanamine hydrochloride (93 mg, 1.36 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (258 mg, 0.68 mmol), and N-diisopropylethylamine (293 mg, 2.27 mmol) in N-dimethylformamide (5 mL) was stirred at room temperature for 2 h. The mixture was then poured into water (10 mL) and extracted with ethyl acetate (10 mL). The organic layer was washed with brine (10 mL), dried over _{Na2SO4 ,} and concentrated. The residue was purified by column chromatography on silica gel (dichloromethane:methanol=30:1) to afford intermediate I143 (25 mg, yield 13%, purity 90% by LCMS) as a yellow oil. LCMS (ESI): Rt= _1.37 min, mass, calcd. For _{C23H30N6O4 454.2} , mass/charge, found value 455.4 [M ₊ H]+.

A mixture of intermediate I143 (25 mg, 0.055 mmol) in hydrochloride/ether (1 mL) was stirred at room temperature for 3 h. The mixture was concentrated in vacuo to give intermediate I144 (25 mg, 100% yield, 90% purity by LCMS) as a yellow oil. LCMS (ESI): R _t = 0.64 min, mass, calcd. For C ₁₈ H ₂₂ N ₆ O ₂ 354.2, mass/charge, found 355.4 [M+H] ⁺ .

A mixture of intermediate I144 (25 mg, 0.07 mmol), 3,4-dichlorobenzoic acid (14 mg, 0.07 mmol), 2-(7-azabenzotriazol-1-yl)-N,N',N'-tetramethyluronium hexafluorophosphate (53 mg, 0.14 mmol), N-diisopropylethylamine (45 mg, 0.35 mmol) in N,N-dimethylformamide (1 mL) was stirred at room temperature for 3 h. The reaction mixture was treated with water (10 mL) and extracted with ethyl acetate (10 mL). _The organic layer was dried over _Na2SO4 and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by preparative HPLC (ACN/ _H2O =50%) to afford compound C79 (10 mg, 27% yield, 99.7% purity by LCMS) as a white solid.

3.2.2.24. Synthesis of C85*R and C85*S compounds

Tert-butyl (1-(4-(difluoromethoxy)phenyl)ethyl)(2-hydroxyethyl)carbamate (500 mg, 1.62 mmol), I101 (642 mg, 1.94 mmol) in dry toluene (2 mL) were charged into a vial. The mixture was purged with nitrogen for 2 min. Cyanomethylenetributylphosphorane (2.1 mL, 1 M, 2.10 mmol) was added and the vial was capped. The reaction mixture was heated at 100 °C for 16 h. The mixture was concentrated in vacuo and the residue was purified by column chromatography (heptane/EtOAc from 100/5 to 50/50) to give 5-(tert-butyl)-3-ethyl-(6R)-2-(2-((tert-butoxycarbonyl)(1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethyl)-6-methyl 2,4,6,7-tetrahydro-5H-pyrαzolo[4,3-c]pyridine-3,5-dicarboxylate I145 (899 mg, 89% yield) as a pale brown oil.

- 100 047929

A test tube was charged with 5-(tert-butyl)-3-ethyl-(6R)-2-(2-((tert-butoxycarbonyl)(1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethyl)-6-methyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate 1145 (899 mg, 1.44 mmol) in dry 1,4-dioxane (11.3 mL). HCl (3.6 mL, 4 M in dioxane, 14.44 mmol) was added and the mixture was stirred at rt for 16 h. The mixture was concentrated in vacuo. The resulting precipitate was triturated in DIPE, filtered and dried under vacuum to give ethyl (6R)-2-(2-((1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethyl)-6-methyl-4,5,6,7-tetrahydro-2Hpyrazolo[4,3-c]pyridine-3-carboxylate I146 (715 mg, quantitative yield).

Ethyl (6R)-2-(2-((1-(4-(difluoromethoxy)phenyl)ethyl)amino)ethyl)-6-methyl4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I146 (715 mg, 1.44 mmol) in toluene (18 mL) was loaded into a test tube. 1,5,7-triazabicyclo[4.4.0]ec-5-ene (TBD) (602 mg, 4.33 mmol) was added and the vial was capped.

The mixture was stirred at 100 °C for 2 h (complete conversion). The mixture was cooled and washed with water. The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography on a SNAP 25 g cartridge in a Biotage system using a gradient (heptane/[EtOAc-EtOH(3:l) + 2% NH ₄ OH]) from 0/100 to 100/0) to give (3R)-9-(1-(4-(difluoromethoxy)phenyl)ethyl)-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5a]pyrazin-10(7H)-one I147 (430 mg, 79% yield) as an oil.

To a solution of (3R)-9-(1-(4-(difluoromethoxy)phenyl)ethyl)-3-methyl-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one 1147 (338 mg, 0.81 mmol) and 4-chloro-3-cyanobenzoic acid (185 mg, 0.97 mmol) in dry DMF (5.4 mL) were added HATU (460 mg, 1.21 mmol) and DIPEA (0.56 mL, 3.23 mmol) at rt. The reaction mixture was stirred at rt for 16 h. The reaction mixture was poured into water (10 mL) and a pink precipitate formed. The solid was filtered off and the product was purified by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 µm,

50x150 mm, mobile phase: 0.25% _NH4HCO3 in water, _CH3CN ) to give a yellow foam. The obtained product C85 was further purified by SFC with separation of enantiomers. Purification was accomplished by preparative SFC (stationary phase: Chiralcel Diacel OJ 20x250 mm, mobile phase: _CO2 , EtOH+0.4 iPrNH2) to give C85*R (87 mg, 20% yield) and C85*S (79 mg, 18% yield), both as yellowish solids.

3.2.2.25. Synthesis of C119*R and C119*S

Ethyl (R)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I111 (836 mg, 2.24 mmol), tert-butyl but-3-yn-2-yl (2-hydroxyethyl) carbamate I109 (573 mg, 2.69 mmol) in dry toluene (14 mL) were charged into a test tube. The mixture was purged with nitrogen for 2 min. Cyanomethylenetributylphosphorane (0.77 mL, 2.92 mmol) was added and the vial was capped. The mixture was heated at 100°C for 16 h. The mixture was concentrated in vacuo and the residue was purified by column chromatography (heptane/EtOAc from 100/0 to 50/50) to give ethyl (6R)-2-(2-(but-3-yn-2-yl(tert-butoxycarbonyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2Hpyrazolo[4,3-c]pyridine-3-carboxylate I148 (979 mg, 64% yield) as a red oil.

I148 (979 mg, 1.72 mmol) was dissolved in DCM (21 mL) and cooled in an ice bath. Ptoluenesulfonyl azide (1.39 mL, 2.07 mmol) diluted in DCM was added, followed by copper(!) thiophene-2-carboxylate (32.9 mg, 0.17 mmol) and stirred for 1 h. The reaction mixture was filtered, partially concentrated, and

- 101 047929 was triturated and purified by column chromatography (heptane/EtOAc from 100/0 to 0/100) to give ethyl (6R)-2-(2-((tert-butoxycarbonyl)(1-(1-tosyl-1H-1,2,3-triazol-4-yl)ethyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylαtα I149 (1.32 g, quantitative yield) as a white foam.

Ethyl (6R)-2-(2-((tert-butoxycarbonyl)(1-(1-tosyl-1H-1,2,3-triazol-4yl)ethyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine3-carboxylate I149 (1.32 g, 1.72 mmol) in dry acetonitrile (ACN) (15 mL) was charged into a test tube. Piperazine (620 mg, 7.21 mmol) was added and the mixture was stirred at 100 °C for 1 h. The mixture was cooled and concentrated in vacuo. The residue was purified by column chromatography (heptane/EtOAc 100/0 to 0/100) to give ethyl (6R)-2-(2-((1-(1H-1,2,3-triazol-4-yl)ethyl)(tert-butoxycarbonyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I150 (544 mg, 52% yield) as a white foam.

A tube was charged with ethyl (6R)-2-(2-((1-(1H-1,2,3-triazol-4-yl)ethyl)(tert-butoxycarbonyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylate I150 (544 mg, 0.89 mmol) in dry DCM (4 mL). TFA (0.82 mL, 10.7 mmol) was added and the mixture was stirred at rt for 2 h. The mixture was concentrated in vacuo. The residue was partitioned between water and Me-THF and neutralized with Na ₂ CO ₃ . The organic layer was separated, dried over _MgSO4 , filtered and concentrated in vacuo. The product, ethyl (6R)-2-(2-((1-(1H-1,2,3-triazol-4yl)ethyl)amino)ethyl)-5-(4-chloro-3-cyanobenzoyl)-6-methyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine3-carboxylate I151, was used as such in the next step.

I151 (455 mg, 0.62 mmol) in dry toluene (5 mL) was charged into a vial. 1,5,7-triazabicyclo[4.4.0]dec-5-ene (600 mg, 4.31 mmol) was added and the vial was capped. The reaction mixture was stirred at 100 °C for 2 h. The mixture was cooled and washed with water. The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The product was purified by column chromatography (heptane/EtOAc from 100/0 to 0/100). The resulting product was triturated in DIPE, filtered and dried under vacuum to give 5-((3R)-9-(1-(1H1,2,3-triazol-4-yl)ethyl)-3-methyl-10-oxo-1,2,3,4,7,8,9,10-octahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazine-2-carbonyl)-2-chlorobenzonitrile C119 (198 mg, 69% yield). The racemic mixture was purified by preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO2, iPrOH+0.4 iPrNH ₂ ). The two fractions obtained were triturated in DIPE, filtered and dried under vacuum to yield C119*R (50 mg, 17% yield) and C119*S (50 mg, 17% yield).

3.2.2.26. Synthesis of compound C75 and analogues

To a solution of 60 wt % sodium hydride in mineral oil (1.2 g, 20 mmol) in H,H-dimethylformamide (30 mL) was added tert-butyl 10-oxo-3,4,7,8,9,10-hexahydro-pyrido[4',3':3,4]pyrazolo[1,5a]pyrazine-2(1H)-carboxylate (3 g, 10 mmol) at 0 °C. After stirring at 0 °C for 30 min, 2-(bromomethyl)pyridine (2.64 g, 15 mmol) was added to the mixture at 0 °C. The mixture was then stirred at 0 °C under nitrogen atmosphere for 2 h. The mixture was quenched with saturated ammonium chloride (30 mL) and extracted twice with ethyl acetate (100 mL). The combined organic layers were washed with brine (50 mL), dried over _Na2SO4 (solid) and filtered. The filtrate was concentrated in vacuo to give a residue which was purified by C18 column (acetonitrile:water=50-70%) to afford intermediate _I152 (3 g, 90% purity by _1H NMR, 77% yield) as _a yellow oil. LCMS (ESI): Rt=1.46 min, mass calcd for _{C20H25N5O3 383.20} , mass/charge found 384.5 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.55 (d, J=4.4 Hz, 1H), 7.70-7.66 (m, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.247.20 (t, 1H), 4.84 (s, 2H), 4.71 (s, 2H), 4.33-4.30 (t, J=5.6 Hz, 2H), 3.88-3.86 (m, 2H), 3.71 (s, 2H), 2.77 (s, 2H), 1.48 (s, 9H).

A solution of intermediate I152 (1.00 g, 2.6 mmol) in 4 M HCl/EtOAc (20 mL) was stirred at room temperature for 0.5 h. The reaction mixture was concentrated under reduced pressure to give the hydrochloride salt of intermediate I53 (900 mg, 80% purity by 1H NMR, crude) as a yellow solid. LCMS (ESI): R _t = 0.34 min, mass calcd for C ₁₅ H ₁₇ N ₅ O 283.14, mass/charge found 284.3 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) δ 9.73 (s, 2H), 8.80-8.79 (d, J=5.6 Hz, 1H), 8.41 (s, 1H), 7.95 (d, J=4.0 Hz, 1H), 7.86 (s, 1H), 5.02 (s, 2H), 4.49

- 102 047929 (t, J=6.0 Hz, 2H), 4.22 (s, 2H), 4.01-3.98 (m, 2H), 3.38 (s, 2H), 2.95 -2.93 (m, 2H).

A solution of intermediate I153 (100 mg, 0.28 mmol), intermediate I76 (62 mg, 0.28 mmol), 2-(7-azabenzotriazol-1-yl)-X,X,X',X'-tetramethyluronium hexafluorophosphate (175 mg, 0.42 mmol) and N,N-diisopropylethylamine (115 mg, 0.84 mmol) in N,N-dimethylformamide (5 mL) was stirred at room temperature for 2 h. The mixture was poured into water (20 mL) and extracted twice with _ethyl acetate (20 mL). The combined organic layers were washed with brine (50 mL), dried over _Na2SO4 (solid) and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified using a C18 column (acetonitrile:water = 35% to 65%) to give compound C75 (80 mg, purity 99.8%, yield 60%) as a white solid.

The compounds listed in the table below are similar

m.

Connection No. Structure Reagents Exit C71 O 0 1. Intermediate 1153 2. 4-Chloro-3-methoxybenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5.DMF 27% C78 N. 1. Intermediate 1153 2. 4-Chloro-3-fluorobenzoic acid 3.HATU 4. Ν, N-Diisopropylethylamine 5.DMF 24% C72 U (G % o 1. Intermediate 1153 2. 4-Chloro-2-fluorobenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5.DMF 23% C73 ⁰ 1. Intermediate 1153 2. 5,6-Dichloronicotinic acid 3.HATU 4. Ν, N-Diisopropylethylamine 5.DMF 39% C74 C76 U U A A 0 ό 1. Intermediate 1153 2. 4-Chloro-2-methylbenzoic acid 3.HATU 4. Ν, N-Diisopropylethylamine 5.DMF 1. Intermediate 1153 2. 4-Chloro-2-methoxybenzoic acid 3. HATU 4. Ν, N-Diisopropylethylamine 5.DMF 35% 47%

3.2.2.27. Synthesis of compound C77

To a solution of intermediate I106 (100 mg, 95% purity, 0.34 mmol) and 4-chloro-3(difluoromethyl)benzoic acid (76 mg, 0.37 mmol) in \,\-dimethylformamide (5 mL) were added O-(7-azabenzotriazol-1-yl)-H N,N',N'-tetramethyluronium hexafluorophosphate (192 mg, 0.51 mmol) and N,N-diisopropylethylamine (130 mg, 1.01 mmol) at 0°C. After stirring at room temperature,

- 103 047929 under nitrogen atmosphere for 2 h, the mixture was poured into water (30 mL) and extracted three times _with ethyl acetate (30 mL). The combined organic layers were washed with water (30 mL), dried over _Na2SO4 (solid) and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by C18 column (acetonitrile:water = 35% to 65%) to give compound C77 (17 mg, yield 10%, purity 99.1%) as a white solid.

3.2.2.28. Synthesis of compounds C94 and 96, 97, 100-102, 104, 107-117, 124 and 125

1106 C94

I106 (300 mg, 1.00 mmol), _4,5 -dichloro-1-(4-methyl)-1,2-dimethyl ... The gray crystals were collected by filtration to give ((4,5-dichloro-1H-indole-2-carbonyl)-3methyl-9-(pyridin-2-ylmethyl)-1,2,3,4,8,9-hexahydropyrido[4',3':3,4]pyrazolo[1,5-a]pyrazin-10(7H)-one C94 (302 mg, 59% yield). The compounds listed in the table below were prepared similarly.

- 104 047929

C107 , .· ^N · H o 50536-62-4 ζχΛ/ ^0Η Η ο 57 C108 F , ^N · 247564-67-6 F ζ>Α, ^οΗ ρ Η ο 61 C109 A’TGg- P N o 1249870-64-1 _f XxK/ ^oh ' Ν Η ο 60 C110 H o 247564-66-5 F _f -CjTKv ^0H Η ο 65 C111 ,.N. 'Yu--р7·-© 7697-29-2 ci ' 'M l Η ο 43 C112 1737-36-6 ο,^_, ΐGL he F ₃ C''x,A, %-/ Η ο 41 PPE ,N. Fcl· O 1192829-74-5 f ₂ hco^ JJGL ° ⁿ οι-ΛΛ, N\\H o 18 C114 F H O 383132-49-8 CjTV/ ^0H Γ'νΛ FH o 54 C115 ,N, 455-24-3 FjC-^TT^OH o 47 C116 Cl ,N, N o 383133-62-8 Cl _f XxK/ ^{oh -} 'Λι Λ H o 72 C117 Cl ,N, J-JP--O 383133-62-8 Cl CjTYv ^oh F Η O 64 C124 But 185 ^F '-' ^F OH L IJH. O H 35 C125 _F F Ή , ^Ν H o 179 F-X-F OH L VW W-N 0 11

* 1-Propanephosphonic anhydride (1.5 equiv) was used (instead of DECP), Et3N (4 equiv) was replaced by DIPEA and DCM was replaced by DMF.

3.2.2.29. Synthesis of compounds C126 and 123, 127-131

- 105 047929

A solution of I14 (50 mg, 0.132 mmol) in 2-MeTHF (5 mL) was added to a Radley tube containing 3-(1-bromoethyl)benzonitrile [1096159-01-1] (33 mg, 0.158 mmol). Then a solution of benzyltriethylammonium chloride [56-37-1] (3 mg, 0.013 mmol) in distilled water (10 μL) and a solution of NaOH (50% in H ₂ O) [1310-73-2] (348 μL, 1.515 g/mL, 6.59 mmol) were added. The two-phase mixture was vigorously stirred for 1 h at 50 °C and then overnight at room temperature. H2O (500 µl) was added and the biphasic mixture was filtered through HM-N drying cartridges and rinsed with 2-MeTHF (2x6 ml). The filtrate was concentrated and _the residue was purified by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 µm, 30x150 mm, mobile phase: 0.25% _NH4HCO3 in water, _CH3CN ) to afford C126 as a solid.

The compounds listed in the table below were prepared in a similar manner.

- 106 047929

3.2.2.30. Synthesis of compounds C122 and 91, 95, 106 and 120

NaH [7646-69-7] (14 mg, 0.360 mmol, 60%) was added to a solution of I14 (100 mg, 0.24 mmol) in DMF (2 mL) at 0 °C. The mixture was warmed to room temperature and stirred for 30 min before adding 2-(chloromethyl)-6-fluoropyridine [315180-16-6] (0.054 mL, 0.48 mmol). The reaction mixture was stirred at room temperature for 5 h and then diluted with EtOAc and water. The organic layer was separated, washed with aqueous saturated NaHCO ₃ solution, brine (2 x 15 mL), dried over Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by column chromatography (DCM/MeOH 100 to 95:5). The product was co-evaporated with EtOH and dried under vacuum. It was then triturated in _Et2O (6 mL) and washed with EtOH (10 mL) to afford compound C122 (86 mg, 48% yield) as a beige powder.

The compounds listed in the table below were prepared in a similar manner.

3.2.2.30. Synthesis of compound C121

F

To a solution of I14 (150 mg, 0.36 mmol) in toluene (1.2 mL) were added sequentially 2-bromo-5-fluoropyridine [41404-58-4] (53 mg, 0.24 mmol), CuI [7681-65-4] (14 mg, 0.072 mmol), N,N'dimethylethylenediamine [110-70-3] (0.023 mL, 0.216 mmol, 0.819 g/mL) and K2CO3 [584-08-7] (99 mg, 0.52 mmol) under argon atmosphere. The reaction mixture was heated at 120 °C for 20 h. _H2O (20 mL) and EtOAc (20 mL) were added to the reaction mixture, the aqueous layer was separated and extracted with

EtOAc (2x10 ml).

The combined organic layers were washed with brine (10 ml), dried over Na ₂ SO ₄ ,

- 107 047929 was filtered and concentrated in vacuo. The residue was purified by column chromatography (DCM/MeOH from 100:0 to 95:5). The resulting solid was co-evaporated with EtOH, dried in vacuo and recrystallized with EtOAc/EtOH. The solid was collected by filtration to give C121 (130 mg, 72% yield) as a white powder.

3.2.2.31. Synthesis of compounds C118 and 86-88, 90, 92, 93, 98, 99 and 103.

Intermediate compound I154 and analogues

DIPEA [7087-68-5] (0.1 mL, 0.589 mmol, 0.75 g/mL) was added to a solution of intermediate I9 (300 mg, 0.491 mmol) and 1-(3-trifluoromethylphenyl)ethylamine [59382-36-4] (139 mg, 0.736 mmol) in DMEU (3 mL). The reaction mixture was stirred under microwave irradiation at 125 °C for 1 h. The reaction mixture was used in the next step without any purification.

The compounds listed in the table below were prepared in a similar manner.

- 108 047929

1,5,7-Triazabicyclo[4.4.0]dec-5-ene [5807-14-7] (150 mg, 1.08 mmol) was added to a solution of I154 (293 mg, 0.49 mmol) in 1,3-dimethyl-2-imidazolidinone [80-73-9] (3 mL) in a microwave vial. The reaction mixture was heated at 130 °C for 1 h in a microwave oven. Then it was diluted with CH ₃ CN and purified by RP-HPLC (stationary phase: RP Xbridge Prep C18 OBD-10 μm, 50x150 mm, mobile phase: 0.25% NH4HCO ₃ in water, CH ₃ CN). The solid was recrystallized with MeOH (2 mL). The product was filtered, washed with H2O and dried at 50°C under vacuum to afford compound C118 (85 mg, 31% yield).

The compounds listed in the table below were prepared in a similar manner.

- 109 047929

3.2.2.32. Synthesis of compounds C84 and 89 and 105 Synthesis of intermediate compound I164

In a vial in Ы ₂ -atm. I14 (250 mg, 0.659 mmol), 1-(1-iodoethenyl)-4-(trifluoromethyl)benzene [359887- 110 047929

55-1] (393 mg, 1.318 mmol), CuI [7681-65-4] (75 mg, 0.396 mmol), _Cs2CO3 [ _534-17-8 ] (430 mg, 1.318 mmol) were dissolved in toluene [108-88-3] (3 mL). Then N,N'-dimethylethylenediamine [110-70-3] (85 μL, 0.791 mmol, 0.819 g/mL) was added and the reaction mixture was stirred at 80 °C over the weekend. The reaction mixture was quenched with saturated aqueous NH4Cl solution and extracted with EtOAc (3 times). The combined organic layers were dried over _MgSO4 , filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography (heptane/EtOAc 50/50 to 20/80) to give I164 (186 mg, 51% yield, 0.339 mmol) as a white solid.

The intermediate compounds listed in the table below were prepared in a similar manner.

To a vigorously stirred solution of I166 (68 mg, 0.146 mmol) _{in CH2Cl2} (1.5 mL) under N2 was added _Et3SiH [617-86-7] (116 μL, 0.728 mmol, 0.728 g/mL). TFA[1493-13-6] (32 μL, 0.364 mmol, 1.696 g/mL) was added dropwise at 0 °C. The reaction mixture was stirred at room temperature overnight. The crude mixture was poured into aqueous NaOH (1 M) and extracted with DCM (4 times). The combined organic layers were dried over _MgSO4 , filtered and concentrated. The residue was purified by column chromatography (100% EtOAc) to give C84 (34 mg, 28% yield) as a white solid.

The compounds listed in the table below were prepared in a similar manner.

R = H or R = Me

- 111 047929

3.2.2.33. Separation of diastereoisomers.

The diastereoisomers listed in the table below were obtained by SFC separation of the corresponding mixture.

Connection No. Structure Way C88*S ^C 4--'A, ⁰ ΓφζΗ Preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO ₂ , EtOH+0.4 iPrNH ₂ ) C88*R ^α ν-Ά _:ι ΑΑ· ο λ—Ν ,---„ ° TO- Preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO ₂ , EtOH+0.4 iPrNH ₂ ) C123*R Cl ° ZA ⁰ f A- ^F \ F Preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO ₂ , EtOH-iPrOH (5050) + 0.4% iPrNH ₂ ) C123*S Cl ci ₄ d. 33 aa ΛύΛ ^{F F} Λ F Preparative SFC (stationary phase: Chiralpak Diacel AD 20x250 mm, mobile phase: CO ₂ , EtOH-iPrOH (5050) + 0.4% iPrNH ₂ ) C105*R Cl ^C1 '-J '''-/V ^N ¢¢-¾ ⁰ T'F Preparative SFC (stationary phase: Chiralpak Daicel IC 20x250 mm, mobile phase: CO ₂ , EtOH+0.4 iPrNH ₂ ) C105*S ¢.¾¾ fr™ [ ' _N M- A Λ Q -¾, Cl F ^f Preparative SFC (stationary phase: Chiralpak Daicel IC 20x250 mm, mobile phase: CO ₂ , EtOH+0.4 iPrNH ₂ )

- 112 047929

3.2.2.34. Synthesis of Compounds C132 and 133-250 General Procedure X1

114 Reagent 2

X = Br, or C-1, or I

I14 (1 mmol) and reagent 2 (1.2 mmol) were placed in a vial. DMF (1 mL) and a solution of tBuONa (3 mmol, 3.0 equiv) [865-48-5] in THF were added. The reaction mixture was stirred for 16 h at 60 °C. After cooling to ambient temperature, the mixture was quenched with excess acetic acid and evaporated. The residue was dissolved in DMSO, filtered, and the solution was purified by HPLC*.

General procedure X2

114 Reagent 2

X = Вг, or С1, or I

I14 (1 mmol) and reagent 2 (1.3 mmol) were placed in a vial. Dry DMF (1 mL) and a solution of LiHMDS (2 mmol, 2.0 equiv) in THF were then added. The reaction mixture was stirred for 16 h at 60 °C. After cooling to ambient temperature, the mixture was quenched with excess acetic acid and evaporated. The residue was dissolved in DMSO, filtered, and the solution was purified by HPLC*.

*Purification was performed using Agilent 1260 Infinity systems equipped with DAD and mass detector. Preparative column was Waters Sunfire C18 OBD, 100 A, 5 µm, 19 mm x 100 mm, with SunFire C18 Prep Guard cartridge, 100 A, 10 µm, 19 mm x 10 mm. Deionized water (Phase A) and HPLC grade methanol (Phase B) were used as eluents. In some cases, ammonia or TFA were used as an additive to improve separation of the products. In such cases, free bases and salts of the TFA products were formed, respectively.

The compounds listed in the table below were synthesized using Method X1 or Method X2.

Connection No. Structure Way HPLC method Exit (%) C132 CI Til ЙТ 'Nx ⁰ Cl XI RP, MeOH+ammonia, gradient 50-100% 21

- 113 047929

C133 Cl X ¹ K N—d Xn 0 X X ^f4 f XI RP, MeOH+ammonia, gradient 40-90% 19 Ck V|1 '(tR> 0 C134 Ck, Cl 1 fa 0 Xi/ 0 V X-N A /1 N^, XI RP, MeOH+ammonia, gradient 30-80% 32 C135 C136 C137 CL Ck ck Cl 0- Cl Cl 0 fax 0 0 li k* 1 ,K,·· 0 fa 7 xx o Xi/ N /к X ^N 0 V -N X 3—N 0 V. j F ' ^N Xr ^F F Xn a N \ / XI XI X2 RP, MeOH+ammonia, gradient 40-90% RP, MeOH+ammonia, gradient 30-80% RP, MeOH+ammonia, gradient 50-100% 39 And 45 C138 C139 Ck Ck Cl Cl 11 'fa' 0 11 fa 0 χ. fai/ 0 V -_-N /X Xn 0 v Fl X' fl Ox. z^N XI XI RP, MeOH+ammonia, gradient 30-80% RP, MeOH+ammonia, gradient 50-100% 29 40 C140 ck Cl 0 K++-X 0 N X Xn ⁰ X Q+. F XI RP, MeOH+ammonia, gradient 50-100% 43

- 114 047929

C141 o o=( g— / s X2 RP, MeOH+ammonia, gradient 40-90% 41 C142 GI T n g 0 Y-N [ 0 V/^N XI RP, MeOH+ammonia, gradient 40-90% 37 C143 Cl ⁰ Xn ⁰ </- ₀ XI RP, MeOH+ammonia, gradient 30-80% 25 C144 Cl ΧίγΝ^Χ^ ⁰ σ ^N Vz _z ^N JJ nn I XI RP, MeOH+ammonia, gradient 30-80% 27 C145 Cl ^C1 --L ''./VN \ I G N-l o Λ~Ν , 0 '^+ft 0-N XI RP, MeOH+ammonia, gradient 40-90% 29 C146 Cl %++> ⁰ ν_/'Ο 'ум XI RP, MeOH+ammonia, gradient 40-90% 23 C147 Cl Cl-^k, '' yj.L_.n \a _O N/^ \^NX XI RP, MeOH+ammonia, gradient 40-90% 20 C148 Cl uyx^ ⁰ °^~ ^ν μ ^ν τγ XI RP, MeOH+ammonia, gradient 30-80% 14

- 115 047929

C149 Cl ^C| '-.D '-/VN Tn and g ν-λ. o 0 - ⁰ ЧУ Vn XI RP, MeOH+ammonia, gradient 40-90% 4 C150 Cl TDgXd 0 U-NS ⁰ XI RP, MeOH+ammonia, gradient 50-100% 22 C151 CI TDgZz ⁰ M=D XI RP, MeOH+ammonia, gradient 30-80% 16 C152 CI ^S K _.: D 'x/VN 0/:¾ ⁰ LL\ XI RP, MeOH+ammonia, gradient 50-100% 15 C153 CI CI, D . x T I g kddz ° 0¾¾ TD F XI RP, MeOH+ammonia, gradient 50-100% 21 C154 CI o D^ ,—, 0 v_/ \ XI RP, MeOH+ammonia, gradient 60-100% 34 C155 Cl Y fl and T Ν—, LI ν.,..,·-Ο'' \ ΐ ζ- X ⁰ 0 ^Ζ/ W Dm rd F XI RP, MeOH+ammonia, gradient 40-90% 35 C156 Cl ^s, >Dx ''-./v ^N idTld 0 D ^m o DDo XI RP, MeOH+ammonia, gradient 30-80% 33

- 116 047929

C157 Cl ‘iX Ο Η-N о v/АИ XI RP, MeOH+ammonia, gradient 40-90% 14 C158 Cl C+ ,Χ.. '/NJ ¹ T Ί i ^fR · Г N-л XJX-N Λ 0 Λ'Ν _Ν 0 + Ν \ ίΐ ΝΑ 4 XI RP, MeOH+ammonia, gradient 30-80% 13 C159 ο ξ^ο ( °Α -η Α'ξ Lj XI RP, MeOH+ammonia, gradient 50-100% 12 C160 Cl CI--X, '/Ή ^Ν Τ' 11 I T Ν—x Tuc ,.Ν ₄ _.--Τ,;-· \ ⁰ °Ax. XI RP, MeOH+ammonia, gradient 50-100% 16 C161 CI ^C1 aA '-IA-N ΧΑπαΧ^λ o An ^ ^c ' A L XI RP, MeOH+ammonia, gradient 50-100% 19 C162 oh oh, ? / Ж\ Ο _χ УК XI RP, MeOH+ammonia, gradient 50-100% 30 C163 CI ''/VN Х1ДТХ 0 А/ /=. 0 λ__/ \ XI RP, MeOH+ammonia, gradient 50-100% 41 C164 CI ^C| -Ха '' ИА-n ХХаХа 0 Xn /=^ 0 X__/ \ XI RP, MeOH+ammonia, gradient 60-100% 26

- 117 047929

C165 Cl θ N 0 V / ^; N F -. / ^x f ^f XI RP, MeOH+ammonia, gradient 40-90% 44 C166 Cl Ά--¾) _v 0 yj (J \ XI RP, MeOH+ammonia, gradient 50-100% 42 C167 o. <AA AJ o XI RP, MeOH+ammonia, gradient 40-90% 19 C168 Cl CIvA -<^N X\aX ⁿ A o An _ ο F XI RP, MeOH+ammonia, gradient 50-100% 25 C169 Cl ⁰ Aj4 A ^{0 Ρ} Ά F ^F XI RP, MeCN+ammonia, gradient 40-90% 14 C170 Cl ^CI >A '%+Z T Η l ⁷ · Г Ν-ϊ n ' У 0 An ⁰ V-/ \ FF XI RP, MeOH+ammonia, gradient 60-100% 15 C171 Cl ^C| >D X3LCX' ^N 3 ⁰ 4 _n Hf XI RP, MeOH+ammonia, gradient 50-100% 34

- 118 047929

C172 Cl CL 1, o ,Χν θ ν/'-ο f ^a f XI RP, MeOH+ammonia, gradient 40-90% 29 C173 Cl ⁰ _N o \__/у hX / XI RP, MeOH+ammonia, gradient 40-90% 19 C174 Cl ^C %X, T 1 pRJ P ' _N XJk-NК/ 0 JN ⁷ _ 0 V/ \ V ^f 5 XI RP, MeOH+ammonia, gradient 60-100% 31 C175 Cl ^0Ι ·'Ύ^·'Ί ' γ ^Λ '^ ^Ν ХЪАХм I yy ^{0 1v} — У XI RP, MeOH+ammonia, gradient 50-100% 16 C176 o ,,%N ⁰ V^-n-4. №\^J f ff XI RP, MeOH+ammonia, gradient 50-100% 20 C177 Cl ⁰ sGH/^L XI RP, MeOH+ammonia, gradient 50-100% 40 C178 Cl ^Cl yX-, ' N XI HX ChL.L. _x y Λ ,N o Xn Y ^// o V XI RP, MeOH+ammonia, gradient 50-100% 26 C179 Cl ^CI ->A andSNOED ⁰ Chl XI RP, MeOH+ammonia, gradient 30-80% 31

- 119 047929

C180 Cl ^C| \L, LI M. o XL o v/P F XI RP, MeOH+ammonia, gradient 50-100% 20 C181 Cl CK Jx '·. Г il И T 'nx Υτ'ΎΥ f ^{1 >M} D ^F x/— F ^F XI RP, MeOH+ammonia, gradient 60-100% 17 C182 Cl '+w ⁰ sp N-0 XI RP, MeOH+ammonia, gradient 40-90% 53 C183 Cl ⁰ oKa XI RP, MeOH+ammonia, gradient 50-100% 23 C184 / 1' p A / >-z Q O XI RP, MeCN+ammonia, gradient 40-90% 33 C185 ¢1-...,.--¾.. '' _r -''v' ^N 1 ] IW T Ν—, zl.A ..N ₄ /Xy \SG ^:? P\/ ⁰ oDu XI RP, MeOH+ammonia, gradient 50-100% 13 C186 ^Cl vx w 0 W , ^{F 0} YL XI RP, MeOH+ammonia, gradient 60-100% 28 C187 Cl ^Cl wK ⁰ σΥΛΝ X2 RP, MeOH+ammonia, gradient 40-90% 14 C188 ^CI VY -PY-N 11 F Π _;| LAzhr· Λ o Di __/ ^{F 0} X2 RP, MeOH+ammonia, gradient 60-100% 28

- 120 047929

C189 0% -00 η if(R) p ^Cl + --0 ) _F 0 Λ'Ν V ο F X2 RP, MeOH+ammonia, gradient 60-100% 24 C190 01-00000/ ) ⁰ У-N _n ⁰ )Z'N /0 Ji ^N ·χ XI RP, MeOH+ammonia, gradient 40-90% 19 C191 o o yX o=( r 0\ yX Oo XI RP, MeOH+ammonia, gradient 50-100% 21 C192 ί?° 0 y ω o XI RP, MeOH+ammonia, gradient 60-100% 32 C193 ^with 0-<x >, XY fZy-y CI'0yN -> ⁰ y. ⁰ V. + ^N / \ ΐ 0 XI RP, MeOH+ammonia, gradient 40-90% 41 C194 00 _: , I У ν'Χ ^ν λ С1 ' ^: ]f' \ > 0 0Ν 0/ _F °4~ ^f F XI RP, MeOH+ammonia, gradient 60-100% 30 C195 Cl ^CI X '-00 UyCX'^ 0 /~N o %''% XI RP, MeOH+ammonia, gradient 40-90% 9 C196 Cl Cl '- 0V> ^N - G 11 i ^Rl 10/00 F /=< ⁰ Hu F XI RP, MeOH+ammonia, gradient 60-100% 13

- 121 047929

C197 Cl Τ Η Ρ™ Γ ΑΧ. ,ν . ΑΑ А Π \ / ξ ο zAn V, ο Ά '. F XI RP, MeOH+ammonia, gradient 50-100% 28 C198 Cl ^Cl -wk ·,·-··-·.^Ν ICHLN \\ Αο ^ν Ν \ XI RP, MeOH+ammonia, gradient 40-90% 12 C199 CI ~ΐ .O'-л. .....· +Α F XI RP, MeCN+ammonia, gradient 50-100% 18 C200 Cl -<Ά- ^{Ν 0} <η_5» Λ-\ XI RP, MeOH+ammonia, gradient 30-80% 20 C201 CI ^C| aA ^; 'ΑΑ= ^ν Τ fl A Γ Ν—, 0 AL Ν ο '>ν XI RP, MeOH+ammonia, gradient 30-80% 17 C202 CI Τ fl P ^RJ Г 0 ...An 0 '' Fn XI RP, MeOH+ammonia, gradient 30-80% 38 C203 Cl A-Λ -<^N T fl 'M o ,-Sn' _ ο νΑΛ Ct .Af F XI RP, MeOH+ammonia, gradient 50-100% 30 C204 Cl ^CI >A '-.<VN XI CQa 0 λ-ti ⁰ a/A 'A/ ¹ — ^SN XI RP, MeOH+ammonia, gradient 50-100% 15

- 122 047929

C205 .й & X XI RP, MeOH+ammonia, gradient 50-100% 24 C206 Cl ^Cl xA ХХХХъ 0 .An Ο /=\ F XI RP, MeOH+ammonia, gradient 65-85% 14 C207 Cl ^with hA ·<^Ν 0 Y-N ___ 0 XI RP, MeCN+water, gradient 45-70% 10 C208 CI ^C1 %A Τ Η f ^RJ g %>+.., ν A==/ Α, 0 )~Ν ^/ __ 0 'l,' \ XI RP, MeCN+water, gradient 25-50% 14 C209 Cl ^CI '_>i A ''-X^N XJLnXX/ ⁿ ~\ o /bj 0 M'N * I! ⁰ X XI RP, MeCN+HCOOH, gradient 20-40% 15 C210 Cl YufYH o U~G4 _ ο v/\ V/ F XI RP, MeOH+ammonia, gradient 50-100% 12 C211 Cl ^c| -xA θ 4~h/ 0 V/^=\ λ XI RP, MeCN+HCOOH, gradient 25-55% 10 C212 Cl ^ί3 *'^ _Ύί ί^ _> '- ,-\ _-N _F 0 ..AN —/ 0 o ^z F XI RP, MeOH+ammonia, gradient 55-70% 21

- 123 047929

C213 Cl 0 .·/ ^N ,___ o V/A V ^{F f} 5 XI RP, MeOH+ammonia, gradient 50-100% 9 C214 Cl ^ci -'--A '-/v ¹ Shul θ XI RP, MeOH+ammonia, gradient 30-80% 15 C215 Cl ^C, >A I m> _l +:--'Y \ ,N 0 y~N ^/'' 0 i —o' XI RP, MeCN+HCOOH, gradient 35-70% 19 C216 Cl Cl^-k ' \ 11 L 1 LLSH-N λ ^Ύ Λ XI RP, MeCN+HCOOH, gradient 40-70% 16 C217 ^С1 ШШ ' I 2 gХш Cl-^^Y' ¹⁴ \ ο /-N ₀ ⁰ A-УN /Л\ Ij nA/ X2 RP, MeOH+ammonia, gradient 50-100% 19 C218 %-+ ''eV JJ g G i-. O U'P _with ⁰ Shz XI RP, MeCN+ammonia, gradient 50-100% 14 C219 I 2 g G N-z C|ШШу'АШу ⁰ g/Λ ^ν ά ° nu XI RP, MeOH+ammonia, gradient 40-90% 27 C220 Cl S'Shch HaASCHA ⁰ .Kb / S D! XI RP, MeOH+ammonia, gradient 30-80% 17

- 124 047929

C221 o Q MS’ U 4+' XI RP, MeCN+ammonia, gradient 40-90% 19 C222 Cl ++¾ ο _χ ί^ ^Ν , 0,.. 0 ν Ν\^ XI RP, MeCN+HCOOH, gradient 35-60% 10 C223 Ύ Ί |ί™ Γ α· --- γι \ / Ο θ-Μ ⁰ l+l XI RP, MeOH+ammonia, gradient 30-80% 15 C224 Ο Ο fa γζί Μ _ζ γ=^ζ XI RP, MeOH+ammonia, gradient 30-80% 16 C225 ?Υ ν ^ζ angle (ε / fa α <J XI RP, MeOH+ammonia, gradient 30-80% 16 C226 Cl ^c| xA Y) ^|iRi g - ^J _Y ^N -faj λ b'N XI RP, MeCN+HCOOH, gradient 30-55% 10 C227 M vx / .—z Q XI RP, MeOH+ammonia, gradient 50-100% 13 C228 Cl T fl fa Γν-λ O fa~N __f X? V X2 RP, MeOH+ammonia, gradient 60-100% 17 C229 Cl ^C1 ^U>'' XX 0 /A-n _n 0 </ -<) U XI RP, MeCN+ammonia, gradient 30-80% 9

- 125 047929

C230 Cl ^CI >A ULcXa 0 >N 0 \ Vy A XI RP, MeOH+ammonia, gradient 50-100% 14 C231 Cl Cl-,4 C./' N Xjl naAa 0 A ^n? 0 λ__/\ ^F 4 XI RP, MeOH+ammonia, gradient 50-100% 9 C232 Cl ^α Άχ '' 4¾ o >л~ XI RP, MeCN+HCOOH, gradient 15-40% 13 C233 Cl ^Ck A ''.---'-- N А®ь 0 tsLL AV N F ^F XI RP, MeCN+HCOOH, gradient 40-70% 14 C234 Cl ^CI >A '-.^NT π ί^Γ 4.4-,. ...nчА/ λ о An _ς ⁰ hV Vn XI RP, MeCN+HCOOH, gradient 30-60% 12 C235 Cl ^Ck A, \ H < ^(No. g 4 4 d, . ^N 4+ A ο Ά na ⁰ 4i / XI RP, MeCN+HCOOH, gradient 20-45% 9 C236 Cl ^Cl ---A G fa G 0 N o o XI RP, MeCN+ammonia, gradient 40-90% 5

- 126 047929

C237 Cl ULrXs 0 У/ ___ _{_} 0 V/AF »У ^Р XI RP, MeCN+ammonia, gradient 50-100% 7 C238 Cl ^CI >A + />I T fl g ία 0 y~N _ ⁰ A/A φ,ΆΝ F XI RP, MeCN+water, gradient 35-65% 7 C239 Cl ^CI >A '-ZVN T fl ig Г Ν—, AA ν A 0 Mind >△ o Α/Ί1' N^ri XI RP, MeCN+ammonia, gradient 35-60% 11 C240 Cl SK '·. Ν \ |l Rra 1 ΖΑΑα.Αα У ΰ Уф A/ ^ci F XI RP, MeOH+ammonia, gradient 50-100% 9 C241 Cl CkA ^! '· r 11 Г 'Ά АУх/Ч' ) А УУ -- ^Q 0 E-/7} \=N XI RP, MeOH+ammonia, gradient 40-90% 9 C242 Cl /a o UU _N ⁰ / X2 RP, MeCN+HCOOH, gradient 10-40% 21 C243 „A-SU _oV ¹ M _B ^ν ~4α ^f F ^F XI RP, MeCN+water, gradient 35-60% 3 C244 ^CI A/A '/VN IJ g g y 0 Um\ ⁰ / y) XI RP, MeCN+HCOOH, gradient 55-75% 19

- 127 047929

C245 ΤΙ γΧ ^ν λ сг ^<: у ~ \ / ⁰ Χχ/χ XI RP, MeOH+ammonia, gradient 40-90% 28 C246 ΤΙ εΓ'ν-α ο Ln _ν ο Λ XI RP, MeOH+ammonia, gradient 30-80% 13 C247 η ο W g — / з\ χ ⁴ χ _z^J ir XI RP, MeCN+ammonia, gradient 30-80% 13 C248 η ο χ* °® °υ '< ^Η xL Χχ XI RP, MeCN+HCOOH, gradient 55-80% 7 C249 11 g Χ+ ^C1 '/G ~)-Ν ίι ° Μ-ί! XI RP, MeCN+ammonia, gradient 20-45% 8 C250 ΐί ΐ κ« Τ HD ν ,, Α/ \ ^C| K X ° uC ⁿ XI RP, MeOH+ammonia, gradient 50-100% 4

3.2.3. LCMS data for compounds C49-C250

Connection No. Rt (min.) [M+H] ⁺ [M-Η]’ LCMS Method C49 3.74 449 G C50 4.00 451 G C51 3.76 452 G C52 3.66 460 G C53 3.11 449 G C54 4.17 451 G C55*R 3.41 535 N C55*S 3.41 535 N C56 3.41 471 G C57 2.39 501 G C58 4.05 513 N C59 3.51 470 G C60 3.37 471 N C61*S 4.02 552 N C62 2.50 501 G C63 4.16 513 N

- 128 047929

C64 3.08 476 G C65 2.62 471 H C61*R 3.58 553 H C66 4.14 535 H C67 3.44 535 H C68 3.55 485 H C69 3.85 536 H C70 3.38 492 H C71 3.00 452 H C72 2.86 440 H C73 2.90 457 H C74 2.93 436 H C75 2.79 472 H C76 2.31 452 G C77 3.46 486 H C78 3.05 440 H C79 3.00 527 H C80 4.14 487 G C81 4.27 486 G C82 3.84 494 H C83 1.76 470 IN C84 1,19 537 C C85*R 1.96 540 598 [M+CH3COO]’ E C85*S 1.95 540 598 [M+CH3COO]’ E C86 1,14 531 C C87 0.92 473 531 [M+CH3COO]’ C C88 1.09 508 566 [M+CH3COO]’ C C89 1,12 469 C C90 1,19 563 621 [M+CH3COO]’ C C91 10,10 504 A C92 1.09 543 C C93 2,15 483 I C94 1.05 509 507 c C95 10.50 512 A C96 0.98 475 473 c C97 0.97 495 493 c C98 2.29 561 I C99 1.94 484 I

- 129 047929

C100 0.88 484 482 C C101 0.93 477 475 c C102 0.97 493 491 c C103 1.80 485 I C104 1.56 510 508 I C105 1.23 551 c C105*R 2.23 551 609 [M+CH3COO]’ D C105*S 2.24 551 609 [M+CH3COO]’ D C106 9.90 500 A C107 0.94 473 471 C C108 0.92 477 475 C C109 0.95 473 471 c C110 0.94 477 475 c Clll 0.94 450 c C112 0.98 504 c C113 0.92 502 c C114 0.95 473 471 c C115 0.93 470 c C116 0.99 493 491 c C117 0.98 493 491 c C118 1.26 551 c C119*R 1.47 465 J C119*S 1.48 465 J C120 10.90 522 A C121 10.60 474 A C122 10,00 488 A C123*R 2.17 551 E C123*S 2,19 551 E C124 0.97 509 507 C C125 1.05 527 525 C C88*R 2.03 508 D C88*S 2.03 508 D C126 2.05 508 I C127 2.31 567 625 [M+CH3COO]’ D C128 1.95 502 560 [M+CH3COO]’ D C129 2,00 508 566 [M+CH3COO]’ D C130 2.02 518 516 I C131 2.07 513 571 J

- 130 047929

[M+СНзСОО]· C132 1.43 504 F C133 1.34 509 F C134 1DZ 516 F C135 1.26 541 F C136 1.04 488 F C137 1.39 501 F C138 1.33 474 F C139 0.99 484 F C140 1.34 517 F C141 1.23 474 F C142 1.35 488 F C143 1.38 501 F C144 1,22 474 F C145 1.33 474 F C146 1.38 475 F C147 1.26 473 F C148 1.30 475 F C149 1.33 488 F C150 1.32 490 F C151 1.36 475 F C152 1.23 484 F C153 1.33 488 F C154 1.52 501 F C155 1,22 523 F C156 1.31 474 F C157 1.32 487 F C158 1.26 514 F C159 1.51 538 F C160 1.32 490 F C161 1.49 530 F C162 1.35 490 F C163 1.24 524 F C164 1.57 527 F C165 1.25 542 F C166 1.45 517 F C167 1,19 520 F C168 1.49 517 F C169 1.40 528 F C170 1.48 562 F C171 1.44 538 F C172 1.33 528 F C173 1.09 487 F

- 131 047929

C174 1.45 553 F C175 1.47 512 F C176 1.26 577 F C177 1.26 502 F C178 1.33 530 F C179 1.34 503 F C180 1.37 512 F C181 1.44 555 F C182 1.40 510 F C183 1.52 517 F C184 1.36 515 F C185 1,16 484 F C186 1.53 519 F C187 1.25 531 F C188 1.39 501 F C189 1.48 519 F C190 1.45 515 F C191 1.44 534 F C192 1.49 519 F C193 1.24 499 F C194 1.44 567 F C195 1,21 495 F C196 1.51 505 F C197 1,21 506 F C198 1.30 501 F C199 1.39 522 F C200 1.26 475 F C201 1.30 501 F C202 0.89 473 F C203 1.47 560 F C204 1.27 508 F C205 1.32 565 F C206 1.56 501 F C207 1.49 508 F C208 1.37 488 F C209 1.37 474 F C210 1.33 488 F C211 1.34 495 F C212 1.38 553 F C213 1.48 538 F C214 1.29 474 F C215 1.42 524 F C216 1.46 512 F

- 132 047929

C217 1.52 531 F C218 1.43 490 F C219 1.23 499 F C220 1.34 503 F C221 1.38 488 F C222 1.36 503 F C223 1.36 489 F C224 1.29 489 F C225 1.33 489 F C226 1.33 501 F C227 1.51 513 F C228 1.47 505 F C229 1,18 474 F C230 1.48 512 F C231 1.43 512 F C232 1.08 498 F C233 1.42 538 F C234 1.35 504 F C235 1,18 514 F C236 1.23 474 F C237 1.33 538 F C238 1.38 512 F C239 1.30 500 F C240 1.49 522 F C241 1,18 474 F C242 0.95 473 F C243 1.37 623 F C244 1.51 501 F C245 1,18 505 F C246 1,20 485 F C247 1.29 489 F C248 1.51 519 F C249 1.06 488 F C250 0.99 484 F

3.2.4. SFC data

Connection No. Rt (min.) [M+H] ⁺ [M-H]’ SFC Method C85*R 3.04 599 [M+ _iPrNH2 ] 538 SFC_C C85*S 3,4 599 [M+ _iPrNH2 ] 538 SFC_C C105*R 4.9 610 [M+ _iPrNH2 ] 549 SFCA C105*S 5.66 610 [M+ _iPrNH2 ] 549 SFC_A C119*R 5.19 465 465 SFC_D C119*S 5.85 465 465 SFCD C123*R 4.88 511 SFCB C123*S 5.29 511 SFCB C88*R 7.51 508 506 SFCE C88*S 8.45 508 506 SFC_E

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3.2.5. 1H NMR data

Connection No. Description of NMR C49 1HNMR (400 MHz, DMSO-t/b) δ 7.42-7.40 (m, 1H), 7.35-7.32 (m, 1H), 7.25-7.20 (m, 3H) , 6.91-6.90 (m, 2H), 4.78 (s, 1H), 4.61-4.53 (m, 3H), 4.26 (s, 2H), 3.913.88 (m , 1H), 3.73 (s, 3H), 3.64-3.49 (m, 3H), 2.74 (s, 2H), 2.28 (s, 3H). C50 1HNMR (400 MHz, DM SO-0/6) δ 7.56-7.48 (m, 4H), 7.27-7.22 (m, 2H), 6.99-6.80 (m, 2H ), 4.80 (s, 1H), 4.61-4.51 (m, 3H), 4.26 (s, 2H), 3.91 (s, 1H), 3.74 (s, 3H) , 3.65-3.56 (m, 3H), 2.74 (s, 2H). C51 1HNMR (400 MHz, DM SO-0/6) δ 7.65-7.51 (m, 2H), 7.40-7.32 (m, 1H), 7.30-7.19 (m, 2H ), 6.97-6.86 (m, 2H), 4.79 (s, 1H), 4.61-4.51 (m, 3H), 4.26 (s, 2H), 3.973.81 ( m, 1H), 3.73 (s, 3H), 3.66-3.55 (m, 3H), 2.75 (s, 2H). C52 1H NMR (400 MHz, DM SO-0/6) δ 8.11 (s, 1H), 7.90 (s, 1H), 7.66-7.62 (m, 2H), 7.26-7 .21 (m, 1H), 6.91 (s, 2H), 4.80 (s, 1H), 4.61-4.51 (m, 3H), 4.27 (s, 2H), 3, 91 (s, 1H), 3.73 (s, 3H), 3.66-3.55 (m, 3H), 2.75 (s, 2H). C53 1H NMR (400 MHz, DM SO-0/6) δ 7.39-7.36 (m, 1H), 7.26-7.19 (m, 4H), 6.90 (s, 2H), 4 .79 (s, 1H), 4.61-4.51 (m, 3H), 4.25 (s, 2H), 3.89 (s, 1H), 3.73 (s, 3H), 3.643, 57 (m, 3H), 2.73 (s, 2H), 2.28 (s, 3H). C54 1HNMR (400 MHz, DM SO-0/6) δ 7.57-7.49 (m, 3H), 7.43-7.41 (m, 1H), 7.28-7.21 (m, 2H ), 6.97-6.85 (m, 2H), 4.80 (s, 1H), 4.61-4.50 (m, 3H), 4.25 (s, 2H), 3.90 ( s, 1H), 3.73 (s, 3H), 3.66-3.55 (m, 3H), 2.74 (s, 2H). C55*R 1H NMR (400 MHz, CD3OD) δ 7.71-7.69 (m, 2H), 7.44 (d, J=7.6 Hz, 1H), 7.337.20 (m, 2H), 6.92 (s, 2H), 6.26-5.80 (m, 1H), 5.60-5.42 (m, 1H), 4.69-4.37 (m, 4H), 4.31 (s , 2H), 3.79 (s, 3H), 3.73 (s, 2H), 3.26-2.91 (m, 2H). C55*S 1H NMR (400 MHz, CD ₃ OD) δ 7.68-7.66 (m, 2H), 7.41 (d, J=8.0 Hz, 1H), 7.317.17 (m, 2H), 6 .95-6.83 (m, 2H), 6.25-5.77 (m, 1H), 5.59-5.39 (m, 1H), 4.77-4.38 (m, 4H) , 4.28 (s, 2H), 3.76 (s, 3H), 3.69 (s, 2H), 3.21-2.89 (m, 2H). C56 1HNMR (400 MHz, DM SO-0/6) δ 7.61-7.44 (m, 2H), 7.28-7.20 (m, 2H), 6.97-6.81 (m, 2H ), 4.78 (s, 1H), 4.61-4.52 (m, 3H), 4.26 (s, 2H), 3.89 (s, 1H), 3.73 (s, 3H) , 3.65-3.55 (m, 3H), 2.74 (s, 2H).

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C57 1HNMR (400 MHz, DM SO-0/6) δ 7.75-7.67 (m, ZN), 7.39-7.12 (m, ZN), 6.97-6.85 (m, 2H ), 4.81 (s, 1H), 4.61-4.51 (m, 3H), 4.26 (s, 2H), 3.91 (s, 1H), 3.73 (s, 3H) , 3.66-3.52 (m, 3H), 2.75 (s, 2H). C58 1HNMR (400 MHz, CDC1 ₃ ) δ 7.55-7.49 (m, 2H), 7.24 (s, 1H), 7.15-7.10 (m, 2H), 6.86-6, 82 (m, 2H), 5.43-5.38 (m, 1H), 4.79-4.12 (m, 4H), 3.76-3.44 (m, 7H), 3.122.60 ( m, 4H), 1.25 (s, 3H). C59 1HNMR (400 MHz, CDC1 ₃ ) δ 9.16 (dd, J=4.8, 1.2 Hz, 1H), 7.63 (br s, 1H), 7.52 (s, 2H), 7, 51-7.49 (m, 1H), 7.24 (s, 1H), 5.73-5.37 (m, 1H), 5.01-4.69 (m, 3H), 4.384.35 ( m, 3H), 4.02-3.98 (m, 2H), 3.16-2.91 (m, 1H), 2.67 (br s, 1H), 1.25 (s, 3H). C60 1H NMR (400 MHz, _CDC13 ) δ 8.71-8.69 (m, 2H), 7.52 (s, 1H), 7.47 (br s, 1H), 7.25-7.21 (m, 2H), 5.71-5.37 (m, 1H), 5.13-4.87 (m, 2H), 4.71-4.29 (m, 4H), 4.033.88 (m , 2H), 3.16-2.95 (m, 1H), 2.76-2.61 (m, 1H), 1.25 (s, 3H). C61*S 1HNMR (400 MHz, DMSO-t/6) 7.85-7.72 (m, 2H), 7.56-7.44 (m, 1H), 7.33-7.15 (m, 2H), 6.96-6.82 (m, 2H), 6.00-5.85 (m, 0.6H), 5.44-5.29 (m, 0.4H), 4.92 (br s, 0.4H), 4.71-4.45 (m, 3.2H), 4.38-4.17 (m, 2.4H), 3.73 (s, 3H), 3.69-3, 57 (m, 2H), 3.41-3.35 (m, 0.3H), 3.27-3.17 (m, 0.7H), 3.12-3.02 (m, 0.6H), 2.98-2.88 (m, 0.4H). C62 1H NMR (400 MHz, DMSO-t/6) δ 7.79 (d, J=7.6 Hz, 1H), 7.72 (s, 1H), 7.58 (d, J=7.2 Hz , 1H), 7.40-7.13 (m, 3H), 6.93-6.87 (m, 2H), 4.82 (s, 1H), 4.62-4.50 (m, 3H ), 4.28-4.24 (m, 2H), 3.92 (s, 1H), 3.74 (s, 3H), 3.66-3.61 (m, 2H), 3.54 ( s, 1H), 2.77-2.73 (m, 2H). C63 1H NMR (300 MHz, DM SO-0/6, 60 oC) δ 7.78-7.73 (m, 2H), 7.46 (dd, J=8.4, 1.8 Hz, 1H), 7.29 (d, J=7.8 Hz, 2H), 6.98-6.92 (m, 2H), 5.86-5.43 (m, 1H), 4.68-4.58 ( m, 2H), 4.31-4.27 (m, 2H), 3.78 (s, 3H), 3.71-3.68 (m, 2H), 3.19-3.16 (m, 1H), 3.04-2.95 (m, 1H), 2.77-2.62 (m, 1H), 1.83-1.27 (m, 3H), 0.99-0.79 (m, 3H). C64 1H NMR (400 MHz, DMSO-t/6) δ 8.17 (s, 1H), 7.91-7.79 (m, 2H), 7.28-7.20 (m, 2H), 6, 97-6.85 (m, 2H), 4.81 (s, 1H), 4.61-4.52 (m, 3H), 4.33-4.19 (m, 2H), 3.91 ( s, 1H), 3.73 (s, 3H), 3.66-3.61 (m, 2H), 3.54 (s, 1H), 2.75 (s, 2H). C65 1H NMR (400 MHz, CDC1 ₃ ) δ 8.67 (s, 1H), 8.53-8.52 (m, 2H), 7.52 (s, 1H), 7.49 (d, J=8 .0 Hz, 1H), 7.23 (s, 1H), 5.79-5.30 (m, 1H), 4.92-4.89 (m, 1H), 4.77 (br s, 1H ), 4.56-4.20 (m, 4H), 3.93-3.90 (m, 2H), 3.05 (br s, 1H), 2.66 (d, J=15.6Hz, 1H), 1.25 (s, 3H).

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C61*R 1H NMR (400 MHz, CD ₃ OD) 7.72-7.69 (m, 2H), 7.46-7.44 (m, 1H), 7.30-7.23 (m, 2H), 6 .91-6.89 (m, 2H), 5.99-5.56 (m, 1H), 4.95-4.91(m, 1H), 4.67-4.60 (m, 3H) , 4.37-4.30 (m, 2H), 3.78-3.71 (m, 5H), 3.26-3.15 (m, 2H). C66 1H NMR (400 MHz, DM SO-0/6) δ 7.75-7.73 (m, 2H), 7.46-7.05 (m, 6H), 5.46-5.23 (m, 1H), 4.69-4.51 (m, 3H), 4.31 (s, 2H), 4.16-4.10 (m, 1H), 3.72 (s, 2H), 2.94 (d, J=10.0 Hz, 1H), 2.63-2.56 (m, 1H), 1.23-1.13 (m, 3H). C67 1HNMR (400 MHz, CDC1 ₃ ) δ 7.53 (d, J=l.6 Hz, 1H), 7.50 (d, J=8.4 Hz, 1H), 7.457.41 (m, 1H), 7.35-7.31 (m, 1H), 7.24-7.20 (m, 2H), 7.13 (d, J=8.0 Hz, 1H), 6.58 (t, J= 73.6 Hz, 1H), 5.80-5.63 (m, 0.4H), 5.48-5.30 (m, 0.6H), 4.81-4.67 (m, 2, 6H), 4.56-4.29 (m, 3.4H), 3.76-3.71 (m, 2H), 3.14-2.96 (m, 1H), 2.65 (d, J=16.8 Hz, 1H), 1.25 (s, 3H). C68 1HNMR (400 MHz, DMSO-t/6) δ 7.76-7.72 (m, 1H), 7.52 (d, J=10.8 Hz, 1H), 7.42 (t, J=7 .6 Hz, 1H), 7.28-7.13 (m, 3H), 6.93-6.87 (m, 2H), 4.83-4.79 (m, 1H), 4.614.50 ( m, 3H), 4.28-4.23 (m, 2H), 3.91-3.90 (m, 1H), 3.74 (s, 3H), 3.66-3.69 (m, 2H), 3.54-3.52 (m, 1H), 2.79-2.73 (m, 2H). C69 1HNMR (400 MHz, DMSO-t/6) δ 8.43 (s, 1H), 7.75-7.73 (m, 2H), 7.65 (br s, 1H), 7.48-7, 43 (m, 2H), 7.30 (t, J=73.2 Hz, 1H), 5.42-5.22 (m, 1H), 4.79-4.35 (m, 2H), 4 .48-4.08 (m, 4H), 3.86 (br s, 2H), 2.97-2.91 (m, 1H), 1.18-1.12 (m, 3H). C70 1HNMR (400 MHz, DMSO-b/b) δ 8.18 (s, 1H), 7.95-7.93 (m, 2H), 7.46-7.19 (m, 3H), 6.93 -6.87 (m, 2H), 4.85-4.83 (m, 1H), 4.62-4.50 (m, 3H), 4.28-4.24 (m, 2H), 3 .96-3.89 (m, 1H), 3.74-3.73 (m, 3H), 3.67-3.60 (m, 2H), 3.53-3.49 (m, 1H) , 2.782.74 (m, 2H). C71 1H NMR (400 MHz, DMSO-b/b) δ 8.51-8.49 (m, 1H), 7.76-7.75 (m, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.37-7.29 (m, 2H), 7.18 (s, 1H), 7.02 (d, J=8.0 Hz, 1H), 4.78-4.58 (m, 4H), 4.33 (s, 2H), 3.88-3.86 (m, 6H), 3.57 (s, 1H), 2.75 (s, 2H) C72 1HNMR (400 MHz, DMSO-b/b) δ 8.53-8.49 (m, 1H), 7.79-7.75 (m, 1H), 7.62-7.58 (m, 1H) , 7.53-7.47 (m, 1H), 7.39-7.26 (m, 3H), 4.80 (d, J=11.2 Hz, 2H), 4.68 (s, 1H ), 4.45 (s, 1H), 4.36-4.30 (m, 2H), 3.94 (s, 1H), 3.87-3.79 (m, 2H), 3.52- 3.51 (m, 1H), 2.77 (s, 1H), 2.67 (s, 1H). C73 1H NMR (400 MHz, DMSO-b/b) δ 8.52 (s, 2H), 8.27 (s, 1H), 7.79-7.72 (m, 1H), 7.38-7, 21 (m, 2H), 4.79 (s, 2H), 4.70 (s, 1H), 4.59 (s, 1H), 4.36-4.34 (m, 2H), 3.923.73 (m, 3H), 3.64 (s, 1H), 2.76 (s, 2H).

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C74 1H NMR (400 MHz, CDC1 ₃ ) δ 8.56-8.53 (m, 1H), 7.71-7.65 (m, 1H), 7.41-7.28 (m, 1H), 7 .23-7.09 (m, 4H), 5.21-4.77 (m, 4H), 4.34-4.31 (m, 2H), 4.22-3.52 (m, 4H) , 2.96-2.69 (m, 2H), 2.31-2.23 (m, 3H). C75 1HNMR (400 MHz, DMSO-c/6) δ 8.54-8.49 (m, 1H), 8.29-8.26 (m, 1H), 7.90-7.72 (m, 2H) , 7.39-7.08 (m, 3H), 4.83 (s, 1.4H), 4.80 (s, 1.4H), 4.69 (s, 0.6H), 4.60 (s, 0.6H), 4.37-4.30 (m, 2H), 3.98-3.95 (m, 0.6H), 3.88-3.85 (m, 1.4H) , 3.82-3.79 (m, 0.6H), 3.65-3.62 (m, 1.4H), 2.83-2.81 (m, 2H). C76 1H NMR (400 MHz, CDC1 ₃ ) δ 8.55-8.53 (m, 1H), 7.71-7.64 (m, 1H), 7.40-7.28 (m, 1H), 7.26-7.16 (m, 2H), 7.01-6.90 (m, 2H), 5.14-4.98 (m, 1H), 4.86-4.78 (m, 2H ), 4.56-4.30 (m, 4H), 3.90-3.84 (m, 2H), 3.82-3.77 (m, 3H), 3.56-3.48 (m , 1H), 2.91-2.66 (m, 2H). C77 1H NMR (400 MHz, CD ₃ OD) δ 8.52 (m, 1H), 7.84 (m, 1H), 7.76 (m, 1H), 7.687.61 (m, 2H), 7.46 -7.35 (m, 2H), 7.11 (t, J=54.4 Hz, 1H), 5.61-5.42 (m, 1H), 5.004.86 (m, 2H), 4, 76-4.58 (m, 1H), 4.40 (m, 3H), 3.91 (m, 2H), 3.10-3.06 (m, 1H), 2.69 (m, 1H) , 1.27 (m, 3H). C78 1HNMR (400 MHz, DMSO-c/6) δ 8.52 (s, 1H), 7.76-7.67 (m, 2H), 7.564 (d, J=9.2 Hz, 1H), 7, 34-7.29 (m, 3H), 4.77-4.56 (m, 4H), 4.34 (s, 2H), 3.90-3.85 (m, 3H), 3.55 ( s, 1H), 2.75 (s, 2H). C79 1H NMR (400 MHz, CD ₃ OD) δ 8.52-8.47 (m, 1H), 7.82-7.80 (m, 1H), 7.76-7.66 (m, 2H), 7.45-7.33 (m, 3H), 5.12-5.00 (m, 1H), 4.75-4.57 (m, 4H), 4.03-4.02 (m, 1H ), 3.73-3.57 (m, 3H), 3.25-3.13 (m, 2H), 2.86-2.82 (m, 2H), 2.72 (s, 3H). C80 1HNMR (400 MHz, CDC1 ₃ ) δ 7.53-7.49 (m, 2H), 7.41-7.38 (m, 1H), 7.33-7.29 (m, 1H), 7, 26-7.24 (m, 1H), 7.16-7.06 (m, 2H), 5.68-5.42 (m, 1H), 4.82-4.29 (m, 6H), 3.76-3.73 (m, 2H), 3.08-2.98 (m, 1H), 2.67-2.63 (m, 1H), 1.29-1.21 (m, 3H ). C81 1HNMR (400 MHz, CDC1 ₃ ) δ 7.53-7.51 (m, 2H), 7.33-7.31 (m, 1H), 7.27-7.26 (m, 0.5H), 7.25-7.24 (m, 0.5H), 7.09-6.98 (m, 3H), 5.64-5.37 (m, 1H), 4.78-4.28 (m , 6H), 3.66-3.63 (m, 2H), 3.11-2.99 (m, 1H), 2.71-2.60 (m, 1H), 1.25 (s, 3H ). C82 1HNMR (400 MHz, CDC1 ₃ ) δ 7.67-7.64 (m, 2H), 7.53-7.49 (m, 2H), 7.43-7.41 (m, 2H), 7, 26-7.24 (m, 1H), 5.69-5.33 (m, 1H), 4.82-4.31 (m, 6H), 3.68-3.64 (m, 2H), 3.09-2.99 (m, 1H), 2.71-2.63 (m, 1H), 1.25 (s, 3H).

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C83 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 2.11 (quin, J=6.5 Hz, 2 H) 2.72 (t, J=5.9 Hz, 2 H) 3.46-3.51 (m, 2 H) 3.71 (br s, 2 H) 4.32 (t, J=6.8 Hz, 2 H) 4.61 (s, 2 H) 4, 75 (s, 2 H) 7.26 (dd, J=7.5, 4.8 Hz, 1 H) 7.32 (d, J=7.9 Hz, 1 H) 7.39 (dd, J =8.l, 2.0 Hz, 1 H) 7.64 (d, J=l.8 Hz, 1 H) 7.67 (d, J=8.1 Hz, 1 H) 7.73 (td, J=7.7, 1.8 Hz, 1 H) 8.50 (d, J=4.9 Hz, 1 H) C84 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.61 (d, J=7.0 Hz, 3 H) 2.77 (t, J=5.8 Hz, 2 H) 3.39 (ddd, J=13.1, 6.7, 4.6 Hz, 1 H) 3.64-3.86 (m, 3 H) 4.14-4.36 (m, 2 H) 4.73 (br s, 2 H) 5.80-5.91 (m, 1 H) 7.45 (dd, J=8.4, 2.0 Hz, 1 H) 7.56-7.64 (m, 2 H) 7.67-7.75 (m, 4 H) C85*R 1H NMR (400 MHz, DMSO-t76„ 100°C) δ ppm 1.17 (d, J=6.9 Hz, 3 H) 1.54 (d, J=7.1 Hz, 3 H) 2.55 (d, J=16.0 Hz, 1 H) 2.87-2.97 (m, 1 H) 3.35 (ddd, J=13.2, 6.7, 4.7 Hz, 1 H) 3.69 (ddd, J=13.2, 8.2, 4.8 Hz, 1 H) 4.11-4.39 (m, 3 H) 4.44-4.81 (m, 1 H) 4.90-5.27 (m, 1 H) 5.78 (q, J=7.1 Hz, 1 H) 7.09 (t, J=74.2 Hz, 1 H) 7.12-7.19 (m, 2 H) 7.41 (d, J=8.5 Hz, 2 H) 7.72-7.77 (m, 1 H) 7.77-7.82 (m, 1 H) 8.00 (d, J=l.9 Hz, 1 H) C85*S 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 1.17 (br d, J=6.9 Hz, 3 H) 1.56 (br d, J=6.6 Hz, 3 H) 2.55 (br d, J=15.9 Hz, 1 H) 2.89-2.95 (m, 1 H) 3.26-3.37 (m, 1 H) 3.64-3 .78 (m, 1 H) 4.06-4.39 (m, 3 H) 4.42-4.80 (m, 1 H) 4.90-5.29 (m, 1 H) 5.735.85 (m, 1 H) 7.08 (br t, J=74.2 Hz, 1 H) 7.14 (br d, J=8.2 Hz, 2 H) 7.40 (br d, J=7.5 Hz, 2 H) 7.71-7.83 (m, 2 H) 7.97-8.02 ( m, 1 H) C86 1H NMR (400 MHz, DMSO-0/6, 150°C) δ ppm 1.18 (br d, J=6.6 Hz, 3 H) 1.54 (br dd, J=6.9, 3, 4 Hz, 3 H) 2.54-2.61 (m, 1 H) 2.94 (br dd, J=15.5, 5.8 Hz, 1 H) 3.323.43 (m, 1 H) 3 .64-3.73 (m, 1 H) 3.85 (s, 3 H) 4.11-4.19 (m, 1 H) 4.20-4.26 (m, 1 H) 4.31 (br dd, J=17.6, 4.6 Hz, 1 H) 4.67 (br s, 1 H) 5.08 (br dd, J=17.2, 10.3 Hz, 1 H) 5.73 (br d, J=7.3 Hz, 1 H) 7.07-7.19 (m, 3 H ) 7.38 (br d, J=8.l Hz, 1 H) 7.61 (s, 1 H) 7.66 (br d, J=7.9 Hz, 1 H) C87 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 1.18 (d, J=6.8 Hz, 3 H) 1.53 (dd, J=6.9, 3.9 Hz , 3 H) 2.58 (br d, J=16.1 Hz, 1 H) 2.93 (br d, J=5.7 Hz, 1 H) 3.45 (br s, 1 H) 3, 64-3.73 (m, 1 H) 4.12-4.20 (m, 1 H) 4.22-4.28 (m, 1 H) 4.28-4.34 (m, 1 H) 4.67 (br s, 1 H) 5.08 (br s, 1 H) 5.83 (q, J=7.1 Hz, 1 H) 6.22 (br s, 1 H) 7.42 (dd, J=8,l, 1.1 Hz, 1 H) 7.62 (br s, 1 H) 7.67 (d, J =l.8 Hz, 1 H) 7.69-7.72 (m, 1 H) 12.47 (br s, 1 H)

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C88 1H NMR (400 MHz, DMSO-ό/δ, 100°С) δ ppm 1.18 (d, J=7.0 Hz, 3 H) 1.60 (dd, J=7.0, 4.2 Hz , 3 H) 2.58 (d, J=15.8 Hz, 1 H) 2.93 (br d, J=5.7 Hz, 1 H) 3.35-3.46 (m, 1 H) 3.75 (qd, J=8.5, 4.6 Hz, 1 H) 4.17-4.25 (m, 1 H) 4.26-4.29 (m, 1 H) 4.31 ( br d, J=6.2 Hz, 1 H) 4.65 (br s, 1 H) 5.09 (br s, 1 H) 5.78-5.85 (m, 1 H) 7.41 (dd, J=8.3, 1.9 Hz, 1 H) 7.57 (dd, J=8.1, 4.2 Hz, 2 H) 7.67 (dd, J=l.8, 1.1 Hz, 1 H) 7.70 (d, J=8.1 Hz, 1 H) 7.78 (dd, J=8.5 , 2.8 Hz, 2 H) C89 1H NMR (400 MHz, DMSO-r/b, 100°C) δ ppm 1.57 (d, J=7.3 Hz, 3 H) 2.76 (t, J=5.9 Hz, 2 H) 3.32 (ddd, J=13.3, 6.9, 4.7 Hz, 1 H) 3.63-3.81 (m, 3 H) 4.10-4.19 (m, 1 H) 4.22-4.30 (m, 1 H) 4.73 (s, 2 H) 5.82 (q, J=7.0 Hz, 1 H) 7.26-7.33 (m, 1 H ) 7.34-7.40 (m, 4 H) 7.44 (dd, J=8.1, 2.0 Hz, 1 H) 7.66-7.73 (m, 2 H) C90 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 1.13 (d, J=6.8 Hz, 3 H) 1.72 (s, 6 H) 2.54 (br d, J=15.8 Hz, 1 H) 2.86-2.93 (m, 1 H) 3.79-3.85 (m, 2 H) 4.14 (br d, J=16.9 Hz, 1 H) 4.28-4.33 (m, 2 H) 4.50-4.68 (m, 1 H) 4.85-5.02 (m, 1 H) 7.07 (t, J= 74.4 Hz, 1 H) 7.05-7.08 (m, 2 H) 7.36 (dd, J=8.3, 1.9 Hz, 1 H) 7.39-7.42 (m, 2 H) 7.61 (d, J=l.8 Hz, 1 H) 7.65 (d, J=8.l Hz, 1H) C91 1H NMR (400 MHz, DMSO-0/6, 80°C) δ ppm 1.18 (d, J=6.8 Hz, 3 H) 2.59 (d, J=16.0 Hz, 1 H) 2.96 (dd, J=15.9, 5.9 Hz, 1 H) 3.76-3.82 (m, 2 H) 4.26-4.36 (m, 3 H) 4.64- 4.74 (m, 3 H) 4.96-5.20 (m, 1 H) 7.43 (dd, J=8.2, 1.9 Hz, 1 H) 7.47 (d, J= 8.3 Hz, 1 H) 7.69 (d, J=l.9 Hz, 1 H) 7.72 (d, J=8.3 Hz, 1 H) 7.82 (dd, J=8.3, 2.4 Hz, 1 H) 8.42 (d, J=2.3 Hz, 1 H) C92 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 1.20 (d, J=7.0 Hz, 3 H) 1.51-1.58 (m, 3 H) 2.59 (d, J=15.8 Hz, 1 H) 2.94 (br d, J=5.9 Hz, 1 H) 3.26-3.41 (m, 1 H) 3.63-3.74 (m, 1 H) 3.76-3.81 (m, 6 H) 4.09-4.21 (m, 1 H) 4.22-4.40 (m, 2 H) 4,494.80 (m , 1 H) 4.92-5.23 (m, 1 H) 5.72-5.83 (m, 1 H) 6.90-6.98 (m, 3 H) 7.40-7.46 (m, 1 H) 7.66-7.74 (m, 2 H) C93 1H NMR (400 MHz, DMSO-0/6, 100°C) δ ppm 1.19 (d, J=6.8 Hz, 3 H) 1.55-1.61 (m, 3 H) 2.54 -2.64 (m, 1 H) 2.91-2.96 (m, 1 H) 3.26-3.40 (m, 1 H) 3.66-3.78 (m, 1 H) 4 ,10-4.22 (m, 1 H) 4.23-4.38 (m, 2 H) 4.53-4.78 (m, 1 H) 4.97-5.23 (m, 1 H ) 5.79-5.88 (m, 1 H) 7.27-7.34 (m, 1 H) 7.34-7.40 (m, 4 H) 7.41-7.45 (m, 1 H) 7.657.74 (m, 2 H)

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C94 1H NMR (400 MHz, DM SO-0/6, 80°C) δ ppm 1.28 (d, J=6.9 Hz, 3 H) 2.71 (d, J=15.9 Hz, 1 H ) 3.02-3.14 (m, 1 H) 3.81-3.95 (m, 2 H) 4.30-4.44 (m, 2 H) 4.51 (br d, J=17 .4 Hz, 1 H) 4.72-4.87 (m, 2 H) 5.16 (quin, J=6.6 Hz, 1 H) 5.38 (d, J=17.3 Hz, 1 H) 6.86 (d, J=0.8 Hz, 1 H) 7.31 (ddd, J=7.5, 4.8, 1.2 Hz, 1 H) 7.35-7.42 (m, 2 H) 7.44-7.51 (m, 1 H) 7.79 (td, J=7.7, 1.8 Hz, 1 H ) 8.55 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) 11.99 (br s, 1 H) C95 1H NMR (400 MHz, DM SO-0/6) δ ppm 1.09-1.24 (m, 3 H) 2.56-2.66 (m, 1 H) 2.89-3.01 (m , 1 H) 3.79-3.91 (m, 2 H) 4.05-4.21 (m, 1 H) 4.32-4.57 (m, 3 H) 4.744.96 (m, 2 H) 5.24 (br s, 1 H) 7.35-7.51 (m, 3 H) 7.70-7.80 (m, 3 H) C96 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.25 (d, J=6.9 Hz, 3 H) 2.69 (d, J=15.9 Hz, 1 H) 2.99-3.07 (m, 1 H) 3.79-3.91 (m, 2 H) 4.28-4.42 (m, 2 H) 4.49 (br d, J=17, 3 Hz, 1 H) 4.77 (d, J=3.2 Hz, 2 H) 5.16 (quin, J=6.5 Hz, 1 H) 5.38 (d, J=17.3 Hz , 1 H) 6.78-6.85 (m, 1 H) 7.09-7.16 (m, 1 H) 7.16-7.23 (m, 1 H) 7.28 (ddd, J=7.5, 4.9, 1.2 Hz, 1 H) 7.34-7.40 (m, 1 H) 7.41-7.49 (m, 1 H) 7.76 (td, J=7.7, 1.8 Hz, 1 H) 8.52 (ddd, J=4.8, 1.8, 0 .9 Hz, 1 H) 11.79 (br s, 1 H) C97 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.24 (d, J=6.8 Hz, 3 H) 2.68 (br d, J=16.0 Hz, 1 H ) 3.01-3.10 (m, 1 H) 3.80-3.92 (m, 2 H) 4.28-4.41 (m, 2 H) 4.47 (br d, J=17 ,4 Hz, 1 H) 4.78 (s, 2 H) 5.14 (quin, J=6.6 Hz, 1 H) 5.34 (d, J=17.3 Hz, 1 H) 6, 95 (d, J=0.8 Hz, 1 H) 7.20-7.32 (m, 2 H) 7.37 (d, J=7.7 Hz, 1 H) 7.77 (td, J=7.7, 1.8 Hz, 1 H) 8.53 (ddd, J=4.8, 1.8, 0.9 Hz , 1 H) 11.92 (br s, 1 H) C98 1H NMR (400 MHz, DMSO-t/6, 150°C) δ ppm 1.20 (d, J=6.8 Hz, 3 H) 1.58 (dd, J=7.0, 3.7 Hz , 3 H) 2.60 (d, J=16.l Hz, 1 H) 2.96 (dd, J=15.8, 5.9 Hz, 1 H) 3.40 (dddd, J=13, 2, 8.4, 6.8, 4.8 Hz, 1 H) 3.72 (dddd, J=13,l, 8.1, 4.9, 3.0 Hz, 1 H) 4.14- 4.38 (m, 3 H) 4.63-4.75 (m, 1 H) 5.10 (dd, J=17.2, 10.6 Hz, 1 H) 5.73-5.82 (m, 1 H) 7.34 (dd, J=8.1, 3.3 Hz, 2 H) 7.40 (dd, J=8 ,1, 2.0 Hz, 1 H) 7.53 (dd, J=8.6, 2.0 Hz, 2 H) 7.63 (d, J=l,8 Hz, 1 H) 7.68 (d, J=8.1 Hz, 1 H) C99 1H NMR (400 MHz, DMSO-t/6, 150°C) δ ppm 1.20 (dd, J=6.8, 2.6 Hz, 3 H) 1.62 (dd, J=7.0, 3.3 Hz, 3 H) 2.60 (d, J=15.8 Hz, 1 H) 2.97 (br dd, J=16.0, 5.8 Hz, 1 H) 3.58-3 .70 (m, 1 H) 3.81 (dddd, J=13,l, 8.1, 4.7, 3.6 Hz, 1 H) 4.16-4.37 (m, 3 H) 4 .62-4.77 (m, 1 H) 5.08 (dd, J=17.3, 6.1 Hz, 1 H) 5.83 (qd, J=7,l, 2.1 Hz, 1 H) 7.23-7.31 (m, 1 H) 7.35-7.44 (m, 2 H) 7.63 ( t, J=2.0 Hz, 1 H) 7.68 (dd, J=8.l, 3.1 Hz, 1 H) 7.72-7.80 (m, 1 H) 8.51-8 .59 (m, 1 H)

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C100 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.25 (d, J=6.9 Hz, 3 H) 2.69 (d, J=16.0 Hz, 1 H) 3.01-3.09 (m, 1 H) 3.80-3.94 (m, 2 H) 4.29-4.42 (m, 2 H) 4.49 (br d, J=17, 5 Hz, 1 H) 4.78 (d, J=2.2 Hz, 2 H) 5.04-5.17 (m, 1 H) 5.33 (br d, J=17.4 Hz, 1 H) 6.92 (d, J=0.9 Hz, 1 H) 7.23-7.32 (m, 2 H) 7.37 (d, J=7.8 Hz, 1 H) 7.77 (td, J=7.7, 1.8 Hz, 1 H) 7.82 (ddd, J=9.0, 4.6, 0 ,8 Hz, 1 H) 8.53 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) 12.19 (brs, 1 H) C101 1H NMR (400 MHz, DMSO-0/6, 80°C) δ ppm 1.24 (d, J=6.9 Hz, 3 H) 2.68 (d, J=16.0 Hz, 1 H) 3.01-3.08 (m, 1 H) 3.87 (ddd, J=7.0, 5.3, 2.2 Hz, 2 H) 4.30-4.42 (m, 2 H) 4.47 (br d, J=17.5 Hz, 1 H) 4.78 (d, J=2.3 Hz, 2 H) 5.15 (quin, J=6.6 Hz, 1 H) 5.36 (d, J=17.3 Hz, 1 H) 6.92 (s, 1 H) 7.15-7.32 (m, 3 H) 7.37 (d, J=7.8 Hz, 1 H) 7.77 (td, J=7.7, 1.8 Hz, 1 H) 8.53 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) 11.84 (s, 1 H) C102 1H NMR (400 MHz, DMSO-0/6, 80°C) δ ppm 1.25 (d, J=6.9 Hz, 3 H) 2.69 (d, J=16.0 Hz, 1 H) 2.99-3.08 (m, 1 H) 3.86 (ddd, J=7.0, 5.3, 2.2 Hz, 2 H) 4.30-4.41 (m, 2 H) 4.49 (br d, J=17.4 Hz, 1 H) 4.77 (d, J=2.5 Hz, 2 H) 5.14 (quin, J=6.6 Hz, 1 H) 5.36 (d, J=17.3 Hz, 1 H) 6.85 (s, 1 H) 7.21 (dd, J=10.0, 8.9 Hz, 1 H) 7.28 (ddd, J=7.5, 4.8, 1.1 Hz, 1 H) 7.37 (d, J=7.8 Hz, 1 H) 7.44 (ddd, J=8.9, 4.0, 0.8 Hz, 1 H) 7.76 (td, J=7.7, 1.8 Hz, 1 H) 8.53 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) 11.89 (br s, 1 H) C103 1H NMR (400 MHz, DMSO-0/6, 150°C) δ ppm 1.21 (d, J=7.0 Hz, 3 H) 1.68 (dd, J=7.3, 3.3 Hz , 3 H) 2.60 (d, J=16.l Hz, 1 H) 2.97 (dd, J=16.0, 5.8 Hz, 1 H) 3.543.64 (m, 1 H) 3 .84 (ddt, J=12.9, 7.8, 5.1, 5.1 Hz, 1 H) 4.22-4.39 (m, 3 H) 4.62-4.77 (m, 1 H) 5.09 (dd, J=17.3, 8.9 Hz, 1 H) 5.77 (qd, J=7.l, 4.2 Hz, 1 H) 7.40 (dq, J=8.2, 1.1 Hz, 1 H) 7.60-7.72 (m, 2 H) 8.81 (d, J=2.4 Hz, 2 H) 9.09 (s, 1 H) C104 1H NMR (400 MHz, DMSO-0/6, 80°C) δ ppm 1.19 (d, J=6.9 Hz, 3 H) 2.58 (d, J=16.0 Hz, 1 H) 2.94-3.03 (m, 1 H) 3.78-3.91 (m, 2 H) 4.24-4.41 (m, 3 H) 4.74 (s, 3 H) 4, 98-5.39 (m, 1 H) 7.08 (dd, J=7.0, 1.6 Hz, 1 H) 7.23-7.29 (m, 1 H) 7.29-7, 40 (m, 1 H) 7.70-7.79 (m, 2 H) 8.50 (dd, J=4.8, 1.8 Hz, 1 H) 8.54 (s, 1 H) 8.65 (dd, J=7.0, 0.9 Hz, 1 H) C105*R 1H NMR (400 MHz, DMSO-0/6, 150°C) δ ppm 1.21 (d, J=6.8 Hz, 3 H) 1.64 (d, J=7.0 Hz, 3 H) 2.60 (d, J=15.8 Hz, 1 H) 2.97 (dd, J=16.1, 5.9 Hz, 1 H) 3.43 (ddd, J=13.2, 6.8, 4.8 Hz, 1 H) 3.77 (ddd, J=13.1, 8.0, 4.6 Hz, 1 H) 4.23 (dd, J=7.9, 4.8 Hz, 1 H) 4.27-4.37 (m, 2 H) 4.69 (br t, J=6.4 Hz, 1 H) 5.11 (d, J=17.2 Hz, 1 H) 5.86 (q, J=7.l Hz, 1 H) 7.40 (dd, J=8.1, 2.0 Hz, 1 H) 7.57-7.65 (m, 3 H) 7.66-7.73 (m, 3 H)

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C105*S 1H NMR (400 MHz, DM SO-0/6, 150 °C) δ ppm 1.21 (d, J = 6.8 Hz, 3 H) 1.64 (d, J = 7.3 Hz, 3 H) 2.61 (d, J = 16.3 Hz, 1 H) 2.98 (dd, J = 16.0, 6.1 Hz, 1 H) 3.46 (ddd, J = 13.2, 6.6, 4.8 Hz, 1 H) 3.77 (ddd, J = 13.1, 8.1, 5.0 Hz, 1 H) 4.20-4.39 (m, 3 H) 4.71 (br t, J = 6.4 Hz, 1 H) 5.09 (br d, J = 17.2 Hz, 1 H) 5.86 (q, J=7.2 Hz, 1 H) 7.41 (dd, J=8.3, 1.9 Hz, 1 H) 7.59-7.66 (m, 3 H) 7.66-7.73 (m, 3 H) C106 1H NMR (400 MHz, DM SO-0/6, 80°C) δ ppm 1.19 (d, J=6.9 Hz, 3 H) 2.59 (br d, J=16.0 Hz, 1 H) 2.96 (dd, J=16.0, 5.9 Hz, 1 H) 3.74 (t, J=6.2 Hz, 2 H) 3.88 (s, 3 H) 4.26 -4.35 (m, 3 H) 4.53-4.71 (m, 3 H) 4.95-5.23 (m, 1 H) 6.80 (d, J=8.5 Hz, 1 H) 7.44 (dd, J=8.2, 1.3 Hz, 1 H) 7.66-7.71 (m, 2 H) 7.73 (d, J=8.3 Hz, 1 H) 8.17 (s, 1 H) C107 1H NMR (400 MHz, DMSO-b/b, 80°C) δ ppm 1.23 (d, J=6.9 Hz, 3 H) 2.40 (d, J=1.9 Hz, 3 H) 2.66 (d, J=16.0 Hz, 1 H) 3.07 (s, 1 H) 3.77-3.93 (m, 2 H) 4.33 (td, J=6.1, 2.3 Hz, 2 H) 4.43 (br d, J=17.4 Hz, 1 H) 4.75 (s, 2 H) 5.15 (quin, J=6.6 Hz, 1 H) 5.35 (d, J=17.3 Hz, 1 H) 6.84 (s, 1 H) 6.97 (dd, J=10.2, 8.8 Hz, 1 H) 7.20-7.29 (m, 2 H) 7.35 (d, J=7.8 Hz, 1 H) 7.74 (td, J=7.7, 1.8 Hz, 1 H) 8.47-8.54 (m, 1 H) 11.42 (br s, 1 H) C108 1HNMR (400 MHz, DMSO-b/w, 80°C) δ ppm 1.15-1.29 (m, 3 H) 2.65 (br d, J=15.9 Hz, 1 H) 3.06 (br d, J=l,8 Hz, 1 H) 3.77-3.95 (m, 2 H) 4.24-4.38 (m, 2 H) 4.43 (br d, J=17 .9 Hz, 1 H) 4.75 (s, 2 H) 4.95-5.10 (m, 1 H) 5.27 (br d, J=17.3 Hz, 1 H) 6.70- 7.06 (m, 3 H) 7.21-7.30 (m, 1 H) 7.31-7.41 (m, 1 H) 7.68-7.80 (m, 1 H) 8,468.58 (m, 1 H) 12.19 (br s, 1 H) C109 1H NMR (400 MHz, DMSO-b/b, 80°C) δ ppm 1.22 (d, J=6.9 Hz, 3 H) 2.66 (d, J=15.9 Hz, 1 H) 3.00-3.09 (m, 4 H) 3.77-3.92 (m, 2 H) 4.24-4.39 (m, 2 H) 4.44 (br d, J=17.4 Hz, 1 H) 4.75 (s, 2 H) 5.18 (quin, J=6.6 Hz, 1 H) 5.37 (d, J=17.3 Hz, 1 H) 6.72 (ddd, J=10.6, 2.3, 1.1 Hz, 1 H) 6.85 (s, 1 H) 6.96 (dd, J=9.9, 2.3 Hz, 1 H) 7.26 (ddd, J=7.5, 4.9, 1.2 Hz, 1 H) 7.35 (d, J=7.7 Hz, 1 H) 7.74 (td, J=7.7, 1.8 Hz, 1 H) 8.51 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) 11.41 (s, 1 H) CHO 1HNMR (400 MHz, DMSO-b/w, 80°C) δ ppm 1.19-1.30 (m, 3 H) 2.68 (br d, J=15.8 Hz, 1 H) 2.99 -3.15 (m, 1 H) 3.81-3.92 (m, 2 H) 4.26-4.41 (m, 2 H) 4.42-4.57 (m, 1 H) 4 .69-4.87 (m, 2 H) 5.10-5.26 (m, 1 H) 5.36 (br d, J=17.2 Hz, 1 H) 6.76-6.95 ( m, 2 H) 7.01-7.13 (m, 1 H) 7.23-7.33 (m, 1 H) 7.34-7.44 (m, 1 H) 7.72-7.82 (m, 1 H) 8.47-8.61 (m, 1 H) 11.84 (br s, 1 H )

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CHI 1H NMR (400 MHz, DM SO-0/6, 80°C) δ ppm 1.15 (d, J=6.9 Hz, 3 H) 2.37 (s, 3 H) 2.56 (d, J=16.0 Hz, 1 H) 2.93 (dd, J=15.9, 6.0 Hz, 1 H) 3.83 (dd, J=6.9, 5.4 Hz, 2 H) 4.24 (br d, J=17.6 Hz, 1 H) 4.32 (td, J=6.2, 1.6 Hz, 2 H) 4.74 (s, 4 H) 7.21- 7.29 (m, 2 H) 7.34 (d, J=7.8 Hz, 1 H) 7.38 (d, J=2.l Hz, 1 H) 7.46 (d, J=8.1 Hz, 1 H) 7.75 (td, J=7.7, 1.8 Hz, 1 H) 8.51 (ddd, J=4.8, 1.7, 0.9 Hz, 1 H) C112 1H NMR (400 MHz, DM SO-0/6, 80°C) δ ppm 1.17 (d, J=6.8 Hz, 3 H) 2.57 (br d, J=16.0 Hz, 1 H) 2.95 (dd, J=15.9, 5.9 Hz, 1 H) 3.76-3.92 (m, 2 H) 4.16-4.40 (m, 3 H) 4, 41-5.45 (m, 4 H) 7.23-7.30 (m, 1 H) 7.34 (d, J=7.8 Hz, 1 H) 7.70-7.82 (m, 3 H) 7.85 (d, J=l.5 Hz, 1 H) 8.51 (dd, J=4.8, 1.8 Hz, 1 H) PPE 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.16 (d, J=6.9 Hz, 3 H) 2.58 (d, J=16.0 Hz, 1 H) 2.95 (dd, J=15.9, 6.0 Hz, 1 H) 3.84 (dd, J=6.8, 5.5 Hz, 2 H) 4.16-4.41 (m, 3 H) 4.42-5.28 (m, 4 H) 7.07-7.50 (m, 5 H) 7.64 (d, J=1.9 Hz, 1 H) 7.75 (td, J=7.7, 1.8 Hz, 1 H) 8.51 (ddd, J=4.8, 1.8, 0.9 Hz, 1 H) SI4 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.22 (d, J=6.9 Hz, 3 H) 2.64 (br d, J=16.2 Hz, 1 H ) 2.96-3.17 (m, 4 H) 3.77-3.99 (m, 2 H) 4.25-4.54 (m, 3 H) 4.664.89 (m, 2 H) 5 .00-5.19 (m, 1 H) 5.23-5.42 (m, 1 H) 6.71-7.00 (m, 3 H) 7.21-7.31 (m, 1 H ) 7.31-7.48 (m, 1 H) 7.67-7.90 (m, 1 H) 8.46-8.63 (m, 1 H) 11.76 (br s, 1 H) SI5 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.16 (d, J=6.9 Hz, 3 H) 2.57 (br d, J=16.2 Hz, 1 H ) 2.94 (dd, J=16.0, 6.1 Hz, 1 H) 3.84 (dd, J=7.0, 5.3 Hz, 2 H) 4,075.60 (m, 7 H) 7.27 (dd, J=7.5, 4.8 Hz, 1 H) 7.35 (d, J=7.9 Hz, 1 H) 7.63 (d, J=7.9 Hz, 2 H) 7.75 (td, J=7.7, 1.9 Hz, 1 H) 7.80 (d, J=8.0 Hz, 2 H) 8.48-8.55 (m, 1 H) SI6 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.23 (d, J=6.8 Hz, 3 H) 2.67 (d, J=15.9 Hz, 1 H) 2.98-3.06 (m, 1 H) 3.80-3.89 (m, 2 H) 4.28-4.39 (m, 2 H) 4.47 (br d, J=17, 3 Hz, 1 H) 4.75 (d, J=2.4 Hz, 2 H) 5.14 (quin, J=6.6 Hz, 1 H) 5.34 (d, J=17.3 Hz , 1 H) 6.81 (d, J=0.9 Hz, 1 H) 7.09 (dd, J=9.6, 2.1 Hz, 1 H) 7.17 (ddd, J=9.5, 2.1, 0.8 Hz, 1 H) 7.26 (ddd, J=7.5, 4.8, 1.2 Hz, 1 H) 7.32-7.39 (m, 1 H) 7.75 (td, J=7.7, 1.8 Hz, 1 H) 8.51 (ddd, J=4.8, 1.8, 0 ,9 Hz, 1 H) 11.67-12.09 (m, 1 H)

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C117 1H NMR (400 MHz, DMSO-0/6, 80°C) δ ppm 1.22 (d, J=6.9 Hz, 3 H) 2.65 (d, J=15.9 Hz, 1 H) 2.96-3.05 (m, 1 H) 3.76-3.93 (m, 2 H) 4.26-4.39 (m, 2 H) 4.43 (br d, 1=17, 4 Hz, 1 H) 4.66-4.84 (m, 2 H) 4.92-5.11 (m, 1 H) 5.26 (br d, J=17.3 Hz, 1 H) 6 .79 (d, J=3.0 Hz, 1 H) 6.97-7.05 (m, 1 H) 7.06-7.11 (m, 1 H) 7.26 (ddd, 1=7.5, 4.8, 1.2 Hz, 1 H) 7.35 (d, J=7.8 Hz, 1 H) 7.74 (td, 1=7.7, 1.8 Hz, 1 H) 8.50 (ddd, 1=4.8, 1.8, 0.9 Hz, 1 H) 12.27 (br s, 1 H) C118 1H NMR (400 MHz, DMSO-0/6, 100°C) 5 ppm 1.16 (d, J=7.0 Hz, 3 H) 1.60 (dd, 1=7.2, 4.5 Hz , 3 H) 2.56 (d, J=15.6 Hz, 1 H) 2.91 (m, J=5.9 Hz, 1 H) 3.38 (tdd, J=13.4, 13, 4, 6.9, 4.8 Hz, 1 H) 3.70-3.78 (m, 1 H) 4.12-4.22 (m, 1 H) 4.23-4.28 (m, 1 H) 4.28-4.33 (m, 1 H) 4.63 (br s, 1 H) 5.06 (br s, 1 H) 5.80-5.87 (m, 1 H) 7.377.42 (m, 1 H) 7.56-7.70 (m, 6 H) C119*R 1H NMR (400 MHz, DMSO-b/b, 100°C) δ ppm 1.17 (d, J=6.8 Hz, 3 H) 1.55 (d, J=7.1 Hz, 3 H) 2.50-2.65 (m, 1 H) 2.95 (dd, J=15.9, 6.0 Hz, 2 H) 3.40-3.52 (m, 1 H) 3.70 (ddd, J=13.1, 8.1, 4.9 Hz, 1 H) 4.15-4.36 (m, 3 H) 4.60 (s, 1 H) 4.92-5.17 (m, 1 H) 5.89 (q, J=7.1 Hz, 1 H) 7.73-7.78 (m, 2 H) 7.78-7.82 (m, 1 H) 8.00 (d, J=l.9 Hz, 1 H) C119*S 1H NMR (400 MHz, DMSO-b/w, 100°C) δ ppm 1.16 (d, J=6.8 Hz, 3 H) 1.56 (d, J=7.l Hz, 3 H) 2.55 (d, J=16.l Hz, 1 H) 2.85-3.03 (m, 2 H) 3.38-3.50 (m, 1 H) 3.68-3.81 ( m, 1 H) 4.13-4.37 (m, 3 H) 4.60 (s, 1 H) 5.07 (br d, J=11.1 Hz, 1 H) 5.89 (q, J=7.1 Hz, 1 H) 7.73-7.77 (m, 2 H) 7.77-7.80 (m, 1 H) 7.98-8.02 (m, 1 H) C120 1H NMR (400 MHz, DMSO-b/w, 80°C) δ ppm 1.19 (d, J=6.9 Hz, 3 H) 2.58 (d, J=16.l Hz, 1 H) 2.92-2.99 (m, 1 H) 3.59-3.66 (m, 2 H) 3.81 (s, 3 H) 4.15-4.27 (m, 2 H) 4, 34 (br d, J=17.3 Hz, 1 H) 4.51-4.79 (m, 1 H) 4.88-4.97 (m, 2 H) 5.02-5.25 (m , 1 H) 6.53 (d, 1=2.9 Hz, 1 H) 7.03 (d, J=7.1 Hz, 1 H) 7.16 (t, J=7.7 Hz, 1 H) 7.29 (d, J=3.0 Hz, 1 H) 7.40 (d, J=8.1 Hz, 1 H) 7.45 (dd, 1=8.2, 1.6 Hz, 1 H) 7.71 (d, 1=4.6 Hz, 2 H) C121 1H NMR (400 MHz, DMSO-b/b) δ ppm 1.11-1.27 (m, 3 H) 2.54-2.61 (m, 1 H) 2.93-3.05 (m, 1 H) 4.09-4.27 (m, 1 H) 4.41-4.57 (m, 4 H) 5.21-5.31 (m, 1 H) 5.415.50 (m, 1 H ) 7.47 (dd, 1=8.3, 1.9 Hz, 1 H) 7.75 (d, J=8.3 Hz, 1 H) 7.78 (d, 1=1.8 Hz, 1 H) 7.82-7.90 (m, 1 H) 7.93-8.01 (m, 1 H) 8.49 (br s, 1 H) C122 1H NMR (400 MHz, DMSO-b/b) δ ppm 1.09-1.26 (m, 3 H) 2.56-2.63 (m, 1 H) 2.87-3.01 (m, 1 H) 3.88 (br s, 2 H) 4.06-4.23 (m, 1 H) 4.37 (br s, 2 H) 4.43-4.56 (m, 1 H) 4 .61-4.84 (m, 2 H) 5.15-5.51 (m, 1 H) 7.09 (br d, J=6.5 Hz, 1 H) 7.297.41 (m, 1 H ) 7.45 (dd, 1=8.1, 1.0 Hz, 1 H) 7.71-7.79 (m, 2 H) 7.91-8.05 (m, 1 H)

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C124 1H NMR (400 MHz, DM SO-0/6, 80°C) δ ppm 1.23 (d, J=6.8 Hz, 3 H) 2.67 (d, J=15.9 Hz, 1 H ) 2.98-3.05 (m, 1 H) 3.78-3.90 (m, 2 H) 4.27-4.39 (m, 2 H) 4.44 (br d, J=17 .4 Hz, 1 H) 4.69-4.83 (m, 2 H) 5.09 (quin, J=6.6 Hz, 1 H) 5.32 (d, J=17.3 Hz, 1 H) 6.95 (br s, 1 H) 7.07-7.18 (m, 1 H) 7.22-7.56 (m, 3 H) 7.58-7.66 (m, 1 H) 7.74 (td, J=7.7, 1.8 Hz, 1 H) 8.51 (ddd, J=4.8, 1, 8, 0.9 Hz, 1 H) 11.86 (br s, 1 H) C125 1H NMR (400 MHz, DMSO-t/6, 80°C) δ ppm 1.23 (d, J=6.9 Hz, 3 H) 2.67 (d, J=15.9 Hz, 1 H) 2.97-3.05 (m, 1 H) 3.78-3.91 (m, 2 H) 4.27-4.39 (m, 2 H) 4.45 (br d, J=17, 4 Hz, 1 H) 4.68-4.82 (m, 2 H) 4.99-5.15 (m, 1 H) 5.30 (d, J=17.3 Hz, 1 H) 6, 79-6.89 (m, 1 H) 7.16-7.30 (m, 2 H) 7.35 (d, J=7.8 Hz, 1 H) 7.69-7.82 (m, 2 H) 8.51 (ddd, J=4.9, 1.8, 0.9 Hz, 1 H) 12.07 (br s, 1H) C88*R 1H NMR (400 MHz, DMSO-t/6, 100°C) δ ppm 1.18 (d, J=7.0 Hz, 3 H) 1.60 (d, J=7.3 Hz, 3 H) 2.58 (d, J=15.8 Hz, 1 H) 2.92-2.97 (m, 1 H) 3.38-3.46 (m, 1 H) 3.75 (ddd, J= 13.2, 8.1, 4.8 Hz, 1 H) 4.20-4.34 (m, 3 H) 4.66 (br s, 1 H) 5.04 (br s, 1 H) 5 .82 (q, J=7.0 Hz, 1 H) 7.41 (dd, J=8.l, 2.0 Hz, 1 H) 7.58 (d, J=8.l Hz, 2 H) 7.67 (d, J=l.8 Hz, 1 H) 7.70 (d, J=8.l Hz, 1 H) 7. 76-7.81 (m, 2H) C126 1H NMR (400 MHz, DM SO-0/6 +CC14) δ ppm 1.14-1.29 (m, 3 H) 1.54-1.69 (m, 3 H) 2.52-2.61 (m, 1 H) 2.88-2.97 (m, 1 H) 3.27-3.37 (m, 1 H) 3.66-3.78 (m, 1 H) 4.10-4 .33 (m, 3 H) 4.44-4.67 (m, 1 H) 5.26-5.51 (m, 1 H) 5.81-5.96 (m, 1 H) 7.37 (br s, 1 H) 7.51-7.71 (m, 5 H) 7.75-7.82 (m, 1 H) C131 1H NMR (400 MHz, DM SO-0/6 +CC14) δ ppm 1.17-1.27 (m, 3 H) 1.49-1.62 (m, 3 H) 2.53-2.61 (m, 1 H) 2.95 (br d, J=17.0 Hz, 1 H) 3.58-3.74 (m, 2 H) 3.77 (br d, J=5.0 Hz, 3 H) 4.05-4.30 (m, 3 H) 4.38-4.67 (m, 1 H) 5.25-5.50 (m, 1 H) 5.76-5.93 ( m, 1 H) 6.77-6.95 (m, 3 H) 7.20-7.28 (m, 1 H) 7.38 (br d, J=6.6 Hz, 1 H) 7.557.66 (m, 2 H) C140 1H NMR (400 MHz, DM SO-0/6 +CC14) δ ppm 1.23 (br d, J=7.0 Hz, 3 H) 2.53-2.61 (m, 1 H) 2.95 (br dd, J=16.2, 5.6 Hz, 1 H) 3.64-3.73 (m, 2 H) 3.84 (s, 3 H) 4.064.22 (m, 1 H) 4 .23-4.30 (m, 2 H) 4.37-4.68 (m, 3 H) 5.12-5.51 (m, 1 H) 6.97-7.15 (m, 3 H ) 7.38 (br d, J=7.4 Hz, 1 H) 7.55-7.65 (m, 2 H) C157 1H NMR (400 MHz, DM SO-0/6 +CC14) δ ppm 1.17-1.28 (m, 3 H) 2.23 (s, 3 H) 2.53-2.61 (m, 1 H) 2.94 (br dd, J=16.2, 5.4 Hz, 1 H) 3.64-3.76 (m, 5 H) 4.26 (br s, 3 H) 4.33- 4.57 (m, 3 H) 5.18-5.46 (m, 1 H) 5.90 (br s, 1 H) 7.37 (br d, J=8.3 Hz, 1 H) 7 .59 (s, 1 H) 7.63 (d, J=8.3 Hz, 1 H)

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C160 1H NMR (400 MHz, DMSO-t/b, 121°С) δ ppm 7.69 (d, J=8.2 Hz, 1 Η), 7.64 (d, J=l.9 Hz, 1 Η ), 7.57 (br s, 1 Η), 7.40 (dd, J=8.2, 2.0 Hz, 1 Η), 5.08 (br d, J=17.2 Hz, 1 H ), 4.75-4.86 (m, 2 H), 4.59-4.72 (m, 1 H), 4.26-4.34 (m, 3 H), 3.74-3, 80 (m, 2 H), 2.96 (dd, J=15.9, 5.9 Hz, 1 H), 2.55-2.63 (m, 4 H), 1.18 (d, J=6.9 Hz, 3 H) C166 1H NMR (400 MHz, DMSO-i/6 +CC14) δ ppm 1.13-1.24 (m, 3 H) 2.50-2.57 (m, 1 H) 2.92 (br dd, J =15.4, 5.8 Hz, 1 H) 3.55-3.67 (m, 2 H) 3.85 (br s, 3 H) 4.04-4.25 (m, 3 H) 4 .44-4.77 (m, 3 H) 5.23-5.46 (m, 1 H) 6.70 (br t, J=8.5 Hz, 1 H) 6.79 (br d, J =8.7 Hz, 1 H) 7.27 (q, J=7.5 Hz, 1 H) 7.36 (br d, J=7.5 Hz, 1 H) 7.57 (br s, 1 H) 7.61 (brd, J=8.3 Hz, 1 H) C175 1H NMR (500 MHz, DMSO-ό/δ) δ ppm 1.04-1.24 (m, 3 H) 2.55-2.61 (m, 1 H) 2.89-2.97 (m, 1 H) 3.73-3.87 (m, 2 H) 4.13-4.21 (m, 1 H) 4.35 (br s, 2 H) 4.43-4.52 (m, 1 H) 4.78 (br s, 2 H) 5.32-5.46 (m, 1 H) 7.44 (br d, J=8.1 Hz, 1 H) 7.50-7.75 ( m, 4 H) 7.84-7.91 (m, 1 H) C204 1H NMR (400 MHz, DMSO-b/b +CC14) δ ppm 1.22 (br s, 3 H) 2.52 (br s, 3 H) 2.54-2.62 (m, 1 H) 2 .91-2.98 (m, 1 H) 3.72 (br s, 2 H) 4.30 (br s, 3 H) 4.71 (br s, 3 H) 5.26-5.49 ( m, 1 H) 7.26-7.43 (m, 3 H) 7.56-7.66 (m, 3 H) C221 1HNMR (400 MHz, CHLOROFORM-d) δ ppm 1.28 (br s, 3 H) 1.65 (br s, 3 H) 2.32 (br s, 3 H) 2.65-2.75 (m , 1 H) 2.98-3.19 (m, 1 H) 3.58-3.70 (m, 1 H) 3.71-3.81 (m, 1 H) 4.33 (br s, 3 H) 4.44-4.64 (m, 1 H) 5.34-5.56 (m, 1 H) 5.95-6.08 (m, 1 H) 6.70 (br d, J =4.6 Hz, 1 H) 7.28-7.31 (m, 1 H) 7.48-7.59 (m, 2 H)

Compounds of formula (Ia) were prepared using the following methods. Scheme 1

(Va) (Via) Vos (Vila)

According to Scheme 1, a commercially available or synthetically obtained compound of formula (Va), wherein R ^2a is H or C _1-6 alkyl, is coupled with commercially available or synthetically obtained ethyl 2-(methylaminomethyl)prop-2-enoate under coupling conditions to form an amide bond to yield a compound of formula (VIa). For example, an acidic compound of formula (Va) is reacted with ethyl 2-(methylaminomethyl)prop-2-enoate in the presence of a drying agent such as hydroxybenzotriazole (HOBt)/1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC), 1,1'-carbonyldiimidazole (CDI), benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5b]pyridinium 3-oxide hexafluorophosphate (HATU), 1-hydroxy-7-azabenzotriazole (HOAT), propylphosphonic anhydride (T3P), a suitably selected base such as N,N-diisopropylethylamine (DIPEA), triethylamine (TEA) and the like, in a solvent such as toluene, acetonitrile (ACN), ethyl acetate (EtOAc), dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or a mixture thereof, to obtain a compound of formula (VIa).

A compound of formula (VIa), wherein R ^2a is H or C1-6 alkyl, is subjected to a cyclization reaction using a base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in a solvent such as ACN, at a temperature of about 40-60°C, for a period of 1-3 hours. Subsequent saponification using a base such as sodium hydroxide, potassium hydroxide or the like; in a polar solvent such as water, methanol (MeOH) or a mixture thereof; provides a compound of formula (Vila), wherein R ^2a is H or _C1-6 alkyl.

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Scheme 2

According to scheme 2, a commercially available or synthetically prepared compound of formula (IXa), wherein R ^2a is H or C1-6alkyl, is subjected to an alkylation reaction with 3-chloro-2(chloromethyl)prop-1-ene in a suitable solvent such as DMF, etc., in the presence of a base such as K2CO3, etc., at temperatures ranging from 25 to 75°C for a period of 10-24 h to obtain a compound of formula (IXa). A compound of formula (IXa), wherein R ^2a is H or C 1-6alkyl, is reacted with methanamine in a solvent such as ethanol (EtOH) at a temperature of about 80°C for a period of 16 hours to yield a compound of formula (Xa), wherein R ^2a is H or C _1-6alkyl .

Scheme 3

According to scheme 3, a compound of formula (Xa), wherein R ^2a is H or C1-6alkyl, is subjected to an oxidation reaction using oxidation conditions known to those skilled in the art, for example, OsO4 and NaIO4, in a suitable solvent such as THF, etc., at temperatures ranging from 0°C to 15°C, for a period of 10-20 hours, to obtain a compound of formula (XIa). The compound of formula (XIa) is reacted with a Grignard reagent such as phenylmagnesium bromide, in a suitable solvent such as THF, etc., to obtain a compound of formula (XIIa), wherein R ^2a is H or C1-6alkyl, R ^1a is OH, and R ^4a is phenyl.

A compound of formula (XIIa), wherein R ^2a is H or C1-6alkyl, R ^1a is OH, and R ^4a is phenyl, is subjected to a fluorination reaction with a fluorinating agent such as diethylaminosulfur trifluoride (DAST), (diethylamino)difluorosulfonium tetrafluoroborate (XtalFluor®), bis(2-methoxyethyl)aminosulfur trifluoride (Deoxo-Fluor®), etc., in a suitable solvent such as DCM, etc., at temperatures ranging from -78 to 50°C for a period of 2 to 16 hours to obtain a compound of formula (XIIa), wherein R ^2a is H or C1-6alkyl, R ^1a is F, and R ^4a is phenyl.

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Scheme 4

According to Scheme 4, a compound of formula (Viia) is converted to its corresponding amide using conditions for forming an amide bond known to those skilled in the art or described previously. For example, reacting a compound of formula (Vila) with ammonium chloride; PyBOP; an activating agent such as HOBt; in the presence of a base such as DIPEA, etc.; in a suitable solvent provides a compound of formula (Xiiia). Similarly, a compound of formula (Vila) is reacted with N-methoxymethanamine in the presence of T3P and in the presence of a tertiary amine base such as triethylamine, etc.; in a non-protic organic solvent such as THF, etc.; to provide a compound of formula (XVia), wherein R ^2a is H or C16 alkyl.

Treatment of a compound of formula (Xiiia) with dimethylformamide (DMF) and dimethylacetamide (DMA) provides a dimethylaminomethylenecarbamoyl-substituted compound of formula (XVa). Treatment of a compound of formula (Xiiia) with trifluoroacetic anhydride (TFAA) in the presence of an organic base such as triethylamine (TEA) in a polar aprotic solvent such as dichloromethane provides a nitrile of formula (XVia).

The compound of formula (XVia) is converted to a ketone of formula (XViia) by treatment with an organometallic halide such as methyl magnesium bromide in a polar aprotic solvent such as THF. Subsequent treatment of the compound of formula (XViia) with DMF-DMA at a temperature of approximately 70°C provides the compound of formula (XViiia).

Scheme 5

- 148 047929

A triazolyl-substituted compound of formula (XIXa) is prepared by reacting a compound of formula (XVa) with hydrazine hydrate in an alcoholic solvent.

Alternatively, the compound of formula (XVa) is reacted with hydroxylamine hydrochloride in the presence of a base such as triethylamine in the presence of acetic acid to give the oxadiazolyl-substituted compound of formula (XXa).

Scheme 6

The compound of formula (XVIa) is reacted with sodium azide in the presence of ACN, in a polar aprotic solvent such as DMF, to give a compound of formula (XXIa), wherein R ^4a is tetrazolyl.

Scheme 7

Vos

The compound of formula (XVIIIa) is subjected to cyclization reaction in the presence of hydrazine hydrate to obtain a compound of formula (XXIIa). The compound of formula (XXIIa) is subjected to alkylation reaction with an alkylating agent such as C _1-6 αalkyl halide and the like, a base such as NaH and the like, in a suitable solvent such as THF, DMF and the like, to obtain a compound of formula (XXIIb and XXIIc).

Alternatively, a compound of formula (XVIIIa) with hydroxylamine hydrochloride in the presence of a base such as triethylamine in the presence of acetic acid provides a compound of formula (XXIIIa).

Scheme 8

According to scheme 8, the separation into stereoisomers of formula (XXIVa) (which covers compounds of formulas (XIIa), (XIXa), (XXa), (XXIa), (XXIIa), (XXIIb), (XXIIc) and (XXIIIa) is achieved through

- 149 047929 by supercritical fluid chromatography. Cleavage of the BOC protecting group from a compound of formula (XXIVa) (which encompasses compounds of formulae (XIIa), (XIXa), (XXa), (XXIa), (XXIIa), (XXIIb), (XXIIc) and (XXIIIa), wherein R ^1a , R ^2a and R ^4a are as defined above; is achieved according to procedures known to those skilled in the art and using conventional techniques such as those described in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley & Sons, 1999. For example, under acidic conditions such as TFA/CH ₂ Cl ₂ , HCl/dioxane, etc., a compound of formula (XXVa) is provided.

Subsequent reaction with a commercially available or synthetically obtained compound of formula (XXVIa), wherein R ^1a , R ^2a and R ^4a are as defined above; a suitable base such as TEA, etc.; in a suitable solvent such as DCM, etc.; provides a compound of formula (Ia).

The compounds of formula (Ia) can be converted into their corresponding salts using methods known to those skilled in the art. For example, an amine of formula (Ia) is treated with trifluoroacetic acid, HCl or citric acid in a solvent such as Et2O, CH2Cl2, THF, MeOH, chloroform or isopropanol to obtain the corresponding salt form. Alternatively, trifluoroacetic acid or formic acid salts are obtained by purification conditions using reversed-phase HPLC. Crystalline forms of pharmaceutically acceptable salts of the compounds of formula (Ia) can be obtained in crystalline form by recrystallization from polar solvents (including mixtures of polar solvents and aqueous mixtures of polar solvents) or from non-polar solvents (including mixtures of non-polar solvents).

If the compounds according to the present invention have at least one chiral center, they may accordingly exist as enantiomers. If the compounds have two or more chiral centers, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are within the scope of the present invention.

Compounds of the formulas shown in the schemes above, which are represented as a stereomeric mixture (meaning a mixture of two or more stereoisomers and includes enantiomers, diastereomers, and combinations thereof), are separated by SFC separation.

General procedures.

The following specific examples are provided to further illustrate the present invention and various preferred embodiments. In preparing the compounds described in the examples below and the corresponding analytical data, the following experimental and analytical protocols were followed unless otherwise noted.

Unless otherwise stated, reaction mixtures were stirred with a magnetic stirrer at room temperature (rt ₎ under nitrogen. If solutions were dried, they were typically dried over a drying agent such as _Na2SO4 or _MgSO4 . If mixtures, solutions, and extracts were concentrated, they were typically concentrated on a rotary evaporator under reduced pressure.

Normal phase silica gel chromatography (FCC) was performed on silica gel (SiO2) using pre-packaged cartridges.

Preparative reversed-phase high-performance liquid chromatography (RP HPLC) was performed by any of the following methods:

Method A. Gilson GX-281 semi-preparative HPLC with Phenomenex Synergi C18 (10 μm, 150x25 mm) or Boston Green ODS C18 (5 μm, 150x30 mm) and a mobile phase of 5-99% ACN in water (with 0.225% FA) for 10 min and then hold at 100% ACN for 2 min at a flow rate of 25 ml/min; or

Method B. Gilson GX-281 semi-preparative HPLC with Phenomenex Synergi C18 (10 μm, 150x25 mm) or Boston Green ODS C18 (5 μm, 150x30 mm) and a mobile phase of 5-99% ACN in water (0.1% TFA) for 10 min and then hold at 100% ACN for 2 min at a flow rate of 25 ml/min, or

Method C. Gilson GX-281 semi-preparative HPLC with Phenomenex Synergi C18 (10 μm, 150x25 mm) or Boston Green ODS C18 (5 μm, 150x30 mm) and a mobile phase of 5-99% ACN in water (0.05% HCl) for 10 min and then hold at 100% ACN for 2 min at a flow rate of 25 mL/min, or

Method D. Semi-preparative HPLC Gilson GX-281 with Phenomenex Gemini C18 (10 μm, 150x25 mm), AD (10 μm, 250 mmx30 mm) or Waters XBridge C18 (5 μm, 150x30 mm) column, mobile phase 0-99% ACN in water (with 0.05% ammonium hydroxide v/v) for 10 min and then hold at 100% ACN for 2 min at a flow rate of 25 mL/min, or

Method E: Gilson GX-281 semi-preparative HPLC with a Phenomenex Gemini C18 (10 μm, 150x25 mm) or Waters XBridge C18 (5 μm, 150x30 mm) column, mobile phase 5-99% ACN in water (10 mM _{NH4HCO3 )} for 10 min and then hold at 100% ACN for 2 min at a flow rate of 25 mL/min.

Preparative supercritical fluid high performance liquid chromatography (SFC) was performed on either a Thar 80 Preparative SFC System or a Waters 80Q Preparative SFC System from Waters. The ABPR was set at 100 bar to maintain _CO2 under SF conditions, and the flow rate could be confirmed according to the compound characteristics, at a flow rate of

- 150 047929 ranging from 50 to 70 g/min. The column temperature was ambient temperature.

Mass spectra (MS) were acquired on a SHIMADZU LCMS-2020 MSD or Agilent 1200/G6110A MSD using electrospray ionization (ESI) in positive mode unless otherwise stated. Calculated mass corresponds to exact mass.

Nuclear magnetic resonance (NMR) spectra were obtained on Bruker AVIII 400 spectrometers. The definitions for multiplicities are as follows: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, br=broad. It will be appreciated that for compounds containing an exchangeable proton, the proton may or may not be visible in the NMR spectrum depending on the choice of solvent used to obtain the NMR spectrum and the concentration of the compound in solution.

Chemical names were obtained using ChemDraw Ultra 12.0, ChemDraw Ultra 14.0 (CambridgeSoft Corp., Cambridge, MA), or ACD/Name Version 10.01 (Advanced Chemistry).

Compounds designated as R* or S* are enantiopure compounds where the absolute configuration is not determined.

Intermediate 1a: 2-tert-butyl-8-ethyl-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-Spirido[4',3':3,4]pyrαzolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate

Step A. Ethyl 2-[[tert-butoxycarbonyl(methyl)amino]methyl]prop-2-enoate. To a mixture of tert-butyl N-methylcarbamate (200.00 mg, 1.52 mmol, 1.00 equiv.) in THF (5.00 mL) was added NaH (91.20 mg, 2.28 mmol, 60% purity, 1.50 equiv.) at 0 °C for 0.5 h under N2, then ethyl 2-(bromomethyl)prop-2-enoate (352.10 mg, 1.82 mmol, 1.20 equiv.) was added dropwise to the mixture at 0 °C, and the mixture was stirred at 15 °C for 2 h under N2. The mixture was poured into ice water (10 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (5 mL x 3). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 100/1 to 5/1) to give the title compound (112.00 mg, 460.34 μmol, yield 30.29%) as a colorless oil. ¹ H ЯМР (400 МГц, CDCl ₃ ) δ 6,28 (s, 1H), 5,55 (s, 1H), 4,23 (q, J=7,1 Гц, 2H), 4,07 (br s, 2H), 2,88 (br s, 3H), 1,45 (br s, 9H), 1,31 (br s, 3H).

Step B: Ethyl 2-(methylaminomethyl)prop-2-enoate: To a mixture of ethyl 2-[[tert-butoxycarbonyl(methyl)amino]methyl]prop-2-enoate (112.00 mg, 460.34 μmol, 1.00 equiv) in dioxane (1.00 mL) was added HCl/dioxane (4 M, 5.00 mL, 43.45 equiv) and then the mixture was stirred at 15 °C for 0.5 h. The mixture was concentrated in vacuo to give the title compound (82.50 mg, 459.25 μmol, 99.76% yield, HCl) as a white solid, which was used directly in the next step.

Step C. tert-Butyl 3-[2-ethoxycarbonylallyl(methyl)carbamoyl1-2,4,6,7-tetrahydropyrazolo[4,3-]pyridine-5-carboxylate. A mixture of 5-tert-butoxycarbonyl-2,4,6,7-tetrahydropyrazolo[4,3-c]pyridine-3-carboxylic acid (80.00 mg, 299.31 μmol, 1.00 eq.), ethyl 2-(methylaminomethyl)prop-2-enoate (59.14 mg, 329.24 μmol, 1.10 eq., HCl), _T3P (285.70 mg, 897.93 μmol, 267.01 μL, 3.00 eq.), and TEA (151.44 mg, 1.50 mmol, 207.45 μL, 5.00 eq.) in THF (3.00 mL) was degassed and purged with N2 three times, and the mixture was then stirred at 15 °C for 1 h. for 16 h under N2 atmosphere. The mixture was poured into water (10 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (5 mL x 3). The combined organic phase was washed with brine (10 mL), dried over _{anhydrous Na2SO4} , filtered and concentrated in vacuo. The residue was purified by preparative TLC (DCM/MeOH=10/1) to give the title compound (46.00 mg, 105.49 μmol, yield 35.24%, purity 90%) as a white solid. LCMS: 393 [M+1]. 1I NMR (400 MHz, CDCl ₃ ) δ 6.35 (s, 1H), 5.67 (br s, 1H), 4.63 (s, 4H), 4.18-4.30 (m, 2H), 3.71 (br s, 2H), 2.91-3.47 (m, 3H), 2.74 (br t, 5.4 Hz, 2H), 1.48 (s, 9H), 1.31 (t, J=7.2 Hz, 3H).

Step D. 2-tert-Butyl-8-ethyl-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-nirido[4',3':3,4]-pyrazolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate. A mixture of tert-butyl 3-[2-ethoxycarbonylallyl(methyl)carbamoyl]-2,4,6,7-tetrahydropyrα-zolo[4,3-c]niridine-5-carboxylate (36.00 mg, 91.73 μmol, 1.00 equiv), DBU (6.98 mg, 45.87 μmol, 6.91 μL, 0.50 equiv) in MeCN (1.00 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 50 °C for 2 h under N2. The mixture was poured into water (5 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (3 mL x 3). The combined organic phase was washed with brine (5 mL), dried over anhydrous

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_Na2SO4 , filtered and concentrated in vacuo. The residue was purified by preparative TLC (DCM/MeOH=20/1) to give the title compound (20.00 mg, 50.96 μmol, 55.56 _% yield) as a white solid. LCMS: 393 [M+1].

Intermediate 2a: tert-Butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate ,;HNH m ί

Weight

Step A. 2-(tert-Butoxycarbonyl)-10-methyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-8-carboxylic acid. To a solution of 2-tert-butyl-8-ethyl-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate (intermediate 1, 3.00 g, 7.64 mmol) in MeOH (30.00 mL) was added a solution of NaOH (458.40 mg, 11.46 mmol) in water (6.00 mL). The reaction mixture was stirred at 30 °C for 16 h. The pH of the reaction mixture was adjusted to approximately 6 by adding dilute hydrochloric acid (2 N, 3 mL). The mixture was diluted with water (20 mL) then extracted with ethyl acetate (100 mL x 3). The combined organic phases were dried over anhydrous _{Na2SO4 ,} filtered and the filtrate was concentrated under reduced pressure to give the title compound (2.50 g, crude) as a white solid, which was used directly in the next step without further purification. MS ( ^ESI ): mass calcd for _{C17H24N4O5 , 364.1} ; mass/charge found, 365.1 [M+H] ₊ .

Step B. tert-Butyl 8-carbamoyl-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a mixture of 2-(tert-butoxycarbonyl)-10-methyl11-oxo-2,3,4,7,8,9,10,11-octahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-8-carboxylic acid (200.00 mg, 548.85 μmol) and NH4Cl (146.79 mg, 2.74 mmol, 95.94 μL) in DMF (5.00 mL) were added PyBOP (314.18 mg, 603.73 μmol), HOBt (81.58 mg, 603.73 μmol), and DIPEA (212.80 mg, 1.65 mmol, 287.57 μL). The reaction mixture was stirred at 15 °C for 2 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with dilute HCl (1N, 30 mL×2), and _the organic phases were dried over anhydrous _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol=50 _/ 1 to 10/1) to give the title compound (173.00 mg, 476.05 μmol, yield 86.74%) as a white solid. MS (ESI): mass calcd for _{C17H25N5O4 ,} 363.1; mass/charge, found, 364.1 [M ₊ H] ⁺ .

Step C. tert-Butyl 8-(((dimethylamino)methylene)carbamoyl)-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. tert-Butyl 8-carbamoyl10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (170.00 mg, 467.79 μmol) was dissolved in DMF-DMA (1.80 g, 15.10 mmol, 2.00 mL). The reaction mixture was stirred at 70°C for 16 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (50 mL x 2), and _the organic phases were dried over anhydrous _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure to give the title compound (170.00 mg, crude), which was used directly in the next step without further purification.

Step D. tert-Butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a mixture of tert-butyl 8-(((dimethylamino)methylene)carbamoyl)-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (60 mg, crude) in EtOH (5.00 mL) was added NH2NH2-H2O (14.65 mg, 286.74 μmol, 14.22 μL, purity 98%), then the reaction mixture was stirred at 80°C for 30 min. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (50 mL x 2), and the organic phases were dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM/MeOH=10/1) to give the title compound (30.00 mg, 77.43 μmol, yield 54.01%) as a yellow oil. MS (ESI): mass calcd for C ₁₈ H ₂₅ N ₇ O, 387.2; mass/charge found, 388.2 [M+H] ⁺ .

Intermediate 3a: tert-butyl 10-methyl-8-(1,2,4-oxadiazol-5-yl)-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

- 152 047929

To a mixture of tert-butyl 8-(((dimethylamino)methylene)carbamoyl)-10-methyl-11-oxo-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (80 mg, crude, obtained in step C for intermediate 2) in AcOH (2.00 mL) were added NH ₂ OH-HCl (106.27 mg, 1.53 mmol) and TEA (232.13 mg, 2.29 mmol, 317.99 μL). The reaction mixture was stirred at 50 °C for 3 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (50 mL x 2), and the organic phases were dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (Rf = 0.65, DCM/MeOH = 10/1) to give the title compound (20.00 mg, 51.49 μmol, yield 26.94%) as a yellow oil. MS (ESI): mass calcd for C ₁₈ H ₂₄ N ₆ O ₄ 388.2; mass/charge found 411.2 [M+Na]+.

Intermediate 4a: tert-butyl 10-methyl-P-oxo-8-(III-tetrazol-5-yl)-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

Step A. tert-Butyl 8-cyano-10-methyl-P-oxo-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]|pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a mixture of tert-butyl 8-carbamoyl-10-methyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (Intermediate 2, step B; 500.00 mg, 1.38 mmol) and TEA (696.11 mg, 6.88 mmol, 953.58 μL) in DCM (3.00 mL) was added TFAA (577.94 mg, 2.75 mmol, 382.74 μL) under N2 at 0°C. The reaction mixture was stirred at 15 °C for 3 h. The organic phases were dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , dichloromethane/methanol=60/1-30/1) to give the title compound (320.00 mg, 926.46 μmol, yield 67.13%) as a white solid. MS (ESI): mass calcd for C ₁₇ H ₂₃ N ₅ O ₃ 345.2; mass/charge found 368.2 [M+Na]+.

Step B. tert-Butyl 10-methyl-11-oxo-8-(III-tetrazol-5-yl)-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a mixture of tert-butyl 8-cyano-10-methyl-11oxo-3,4,8,9,10,11-hexahydro-1H-pyrigo[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (80.00 mg, 231.62 μmol) in DMF (2.00 mL) were added _NaN3 (22.59 mg, 347.43 μmol) and ACN (12.70 mg, 23.16 μmol, 11.55 μL) under N2 atmosphere. The reaction mixture was stirred at 110 °C for 16 h. The reaction mixture was diluted with ethyl acetate (50 mL) and washed with dilute HCl (1 N, 30 mL x 2), and the organic phases were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by RP-HPLC (condition A) to give tert-butyl 10-methyl-11-oxo-8-(N-tetrazol-5-yl)-3,4,8,9,10,11-hexahydro-N-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (43.00 mg, 110.70 μmol, 47.80% yield) as a white solid. MS (ESI): mass calcd. for C ₁₇ H ₂₄ N ₈ O ₃ 388.2; mass/charge, found value, 389.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl3) δ 4.82-4.91 (m, 1H), 4.73-4.81 (m, 1H), 4.62 (br s, 2H), 4.19 (br s, 1H), 3.78-3.94 (m, 2H), 3.65 (br s, 2H), 3.20 (br s, 3H), 2.71-2.85 (m, 2H), 1.49 (s, 9H).

Intermediate 5a: tert-butyl 10-methyl-p-oxo-8-(n-pyrazol-3-yl)-3,4,8,9,10,11hexahydro-1H-pyrigo[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

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Step A. tert-Butyl 8-(methoxy(methyl)carbamoyl)-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a mixture of 2-(tert-butoxycarbonyl)-10methyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-1H-pyrido[4',3':3,4]pyrazolo-[1,5-a][1,4]diazepine-8-carboxylic acid (intermediate 2, product from step A; 7.00 g, 19.21 mmol) and N-methoxymethanamine hydrochloride (7.49 g, 76.84 mmol) in THF (100.00 mL) were added _T3P (24.45 g, 38.42 mmol, 22.85 mL, 50% purity) in one portion and TEA (29.16 g, 288.14 mmol) under _N2 . The mixture was stirred at 30 °C for 12 h. The mixture was poured into water (150 mL) and stirred for 15 min, and the aqueous phase was extracted with ethyl acetate (200 mL x 2). The combined organic phases were washed with brine (50 mL x 1), dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH=30/1) to give the title compound (7.50 g, 18.41 mmol, yield 95.82%) as a yellow oil. MS (ESI): mass calcd for C ₁₉ H ₂₉ N ₅ O ₅ 407.2; mass/charge found 408.1 [M+H]+. 1H NMR (400 MHz, CDCl ₃ ) δ 4.69 -4.43 (m, 4H), 3.79-3.73 (m, 1H), 7.56-7.64 (m, 1H), 3.65 3.59 (m, 4H), 3.25 (s, 3H), 3.20 (s, 3H), 70 (m, 2H), 1.48 (s, 9H).

Step B. tert-Butyl 8-acetyl-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-3-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of tert-butyl 8-[methoxy(methyl)carbamoyl]10-methyl-11-oxo-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (300.00 mg, 736.27 μmol) in THF (5.00 mL) was added MeMgBr (3 M, 736.27 μL) dropwise at -30 °C under N ₂ atmosphere. The mixture was warmed to 0 °C and stirred for 1 h. The mixture was slowly poured into HCl (1 N aq, 80 mL) and extracted with EtOAc (50 mL×3). _The combined organic layers were dried over _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure to give the title compound (170.00 mg, crude) as a brown oil. MS (ESI): mass calcd for _{C18H26N4O4 , 362.2} ; mass/charge found, 363.2 [M ₊ H] ⁺ .

Step C. tert-Butyl 8-(3-(dimethylamino)acryloyl)-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-Spyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. tert-Butyl 8-acetyl-10-methyl-11-oxo3,4,8,9,10,11-hexahydro-1H-nirido[4',3':3,4]nirazolo[1,5-a][1,4]diazene-2(7H)-carboxylate (60.00 mg, 165.55 μmol) was dissolved in DMF-DMA (1.35 g, 11.33 mmol, 1.50 mL) and the mixture was stirred at 70 °C for 16 h. The mixture was extracted with EtOAc (20 mL× ₂ ) and water (20 mL). The combined organic layer was dried over _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure to give the title compound (80.00 mg, crude) as a yellow oil. MS (ESI): mass calcd for C ₂₁ H ₃₁ N ₅ O ₄ 417.2; mass/charge found value 418.2 [M+H] ⁺ .

Step D. tert-Butyl 10-methyl-11-oxo-8-(III-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]nirazolo[1,5-a][1,4]diazene-2(7H)-carboxylate. To a solution of tert-butyl 8-(3-(dimethylamino)acryloyl)-10-methyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-p-carboxylate (90.00 mg, 215.57 μmol) in EtOH (500.00 μL) was added NH ₂ NH ₂ -H ₂ O (22.02 mg, 431.14 μmol, 21.38 μL, purity 98%). The mixture was stirred at 80 °C for 2 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (20 mL × 2). The combined organic layers were dried over anhydrous _{Na2SO4 ,} filtered, and the filtrate was concentrated under reduced pressure to give the title compound (70.00 mg, crude) as a yellow oil. MS (ESI): mass calcd for _{C19H26N6O3 , 386.2} ; mass/charge found, 387.1 [M ₊ H] ⁺ .

Intermediate 6a: tert-butyl 10-methyl-8-(1-methyl-III-pyrazol-3-yl)-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]nirazolo[1,5-a][1,4]diazene-2(7H)-carboxylate

Vos

To a solution of tert-butyl 10-methyl-11-oxo-8-(N-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-N-pyrido[4',3':3,4]nirazolo[1,5-a][1,4]diazene-2(7H)-carboxylate (intermediate 5, 90.00 mg, 232.89 μmol) in THF (2.00 mL) was added NaH (27.95 mg, 698.67 μmol, 60% purity) at 0 °C. The mixture was stirred at 0 °C for 30 min. Then CH3I (99.17 mg, 698.67 μmol, 43.50 μL) was added. The mixture was stirred at 20 °C for 2 h. The mixture was quenched by adding saturated NH ₄ Cl (20 mL) and then extracted with EA (30 mL x 2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (EA/MeOH=10/1) to give the title compound (40.00 mg, 99.88 μmol, yield 42.89%, Rf=0.37 (EtOAc/MeOH=10/1)) as a colorless oil, MS (ESI): mass calcd for C ₂₀ H ₂₈ N ₆ O ₃ 400.2; mass/charge found 401.2 [M+H] ⁺ . 1H NMR

- 154 047929 (400 MHz, CDCl ₃ ) δ 7.25 (d, J=2.2 Hz, 1H), 6.07 (d, J=2.0 Hz, 1H), 5.23 (s, 1H ), 4.58-4.50 (m, 3H), 4.48-4.41 (m, 1H), 3.80 (s, 3H), 3.75-3.44 (m, 5H), 3.06 (s, 3H), 2.69 (br, 2H), 1.41 (s, 9H). Regioisomer tert-butyl-10methyl-8-(1-methyl-III-pyrazol-5-yl)-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3' :3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (8.00 mg, 17.48 μmol, yield 7.51%, purity 87.5%, Rf=0 ,30 (EtOAc/MeOH=10/1)) as a colorless oil.

Intermediate 7a: tert-butyl 10-methyl-8-(1-methyl-III-pyrazol-5-yl)-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

The title compound was isolated by preparative TLC from intermediate 5 (8.00 mg, 17.48 μmol, yield 7.51%, purity 87.5%, Rf=0.30 (EtOAc/MeOH=10/1)). MS (ESI): mass calcd for _{C20H28N6O3 , 400.2} ; mass/charge found, 401.2 [M ₊ H]+.

Intermediate 8a: (3R,8S*)-tert-butyl 3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate ζτ

N-N ii f J o V Ν'

Vos

Step A. Ethyl 2-(((tert-butoxycarbonyl)(methyl)amino)methyl)acrylate. To a mixture of NaH (4.57 g, 114.36 mmol, 60% purity) in THF (80.00 mL) was added dropwise tert-butyl N-methylcarbamate (10.00 g, 76.24 mmol) with stirring at -30 °C under N2, followed by ethyl 2-(bromomethyl)prop-2-enoate (19.13 g, 99.11 mmol) after 0.5 h. The resulting mixture was warmed to 20 °C with stirring for 16 h. The mixture was poured into ice water (200 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (200 mL×3). The combined organic phases were washed with brine (200 ml), dried over anhydrous _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography ( _SiO2 , petroleum ether/ethyl _acetate =100/1 to 10/1) to give a mixture of the title compound and ethyl 2(hydroxymethyl)prop-2-enoate (21 g) as a light yellow liquid.

Step B: Ethyl 2-((methylamino)methyl)acrylate hydrochloride: To a mixture of ethyl ethyl 2-(((tert-butoxycarbonyl)(methyl)amino)methyl)acrylate and ethyl 2-(hydroxymethyl)prop-2-enoate (11.5 g, Step A) was added HCl/dioxane (4 M, 30.00 mL) with stirring at 20 °C over 2 h. The mixture was concentrated and the residue was dissolved in water (100 mL). The pH of the resulting mixture was adjusted to 3 by adding HCl (1 N) and the mixture was washed with DCM (100 mL×3). The resulting aqueous phase was concentrated to give the title compound (5.50 g, 30.62 mmol, 64.77% yield, HCl salt) as a colorless liquid.

Step C. (^)-tert-Butyl 3-((2-(ethoxycarbonyl)allyl)(methyl)carbamoyl)-6-methyl-6,7-dihydro-2Hpyrazolo[4,3-c]pyridine-5(4H)-carboxylate. To a solution of ethyl 2-((methylamino)methyl)acrylate hydrochloride (5.50 g, 30.62 mmol) and (N)-5-(tert-butoxycarbonyl)-6-methyl-4,5,6,7-tetrahydro-2Hpyrazolo[4,3-c]pyridine-3-carboxylic acid (5.20 g, 18.48 mmol) in THF (80.00 mL) were added TEA (22.44 g, 221.76 mmol, 30.74 mL) and T3P (58.82 g, 92.42 mmol, 54.97 mL, 50% purity) under N2 atmosphere. The mixture was stirred at 30 °C for 32 h under N2 atmosphere. Additional T3P (23.52 g, 36.96 mmol, 21.98 mL, 50% purity) and TEA (14.96 g, 147.84 mmol, 20.49 mL) were added to the mixture at 30 °C with stirring for another 16 h. The mixture was diluted with 80 mL of water and extracted with EtOAc (50 mL x 4). The organic phase was washed successively with HCl (1 N, 50 mL x 2), saturated NaHCO ₃ and brine (50 mL), then dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0~50% ethyl acetate/petroleum ether gradient) to give the title compound (5.52 g, 13.58 mmol, 73.49% yield) as a white solid. MS (ESI): mass calcd for _{C20H30N4O5 , 406.2} ; mass/charge found, 407.2 [ _M +H] ⁺ .

- 155 047929

Step D. (3R)-2-tert-Butyl-8-ethyl-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate. To a solution of DBU (1.03 g, 6.79 mmol, 1.02 mL) in MeCN (600.00 mL) was added dropwise a solution of ((2-(ethoxycarbonyl)allyl)(methyl)carbamoyl)-6-methyl-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate (5.52 g, 13.58 mmol) in MeCN (150.00 mL) with stirring at 50 °C over 16 h. The mixture was concentrated under reduced pressure and the resulting residue was dissolved in 100 mL of EtOAc. The resulting mixture was then washed with HCl (1N, 100 mL) and brine (100 mL x ₁ ), dried over anhydrous _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography (ethyl acetate/petroleum ether gradient 50~100%) to give the title compound (5.20 g, 12.79 mmol, yield 94.21%) as a colorless oil.

Step E. (3R)-2-(tert-Butoxycarbonyl)-3,10-dimethyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-Spyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-8-carboxylic acid. To a solution of (3R)-2-tert-butyl-8-ethyl-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate (450.00 mg, 1.11 mmol) in THF (10.00 mL) and water (1.00 mL) was added NaOH (88.57 mg, 2.21 mmol), then the mixture was heated to 50 °C for 16 h. The mixture was diluted with 20 mL of water and the pH of the mixture was adjusted from 12 to 5 by adding HCl (1N). The resulting mixture was extracted with EtOAc (20 mL×5). The combined organic phases were then washed with brine (30 mL x 1), dried over _{anhydrous Na2SO4} , filtered, and the filtrate was concentrated under reduced pressure to give the title compound (390.00 mg, 1.03 mmol, yield 92.85%) as a white solid.

Step F. (3R)-tert-Butyl-8-(methoxy(methyl)carbamoyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-2-(tertbutoxycarbonyl)-3,10-dimethyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-N-pyrido-[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-8-carboxylic acid (390.00 mg, 1.03 mmol) and N-methoxymethanamine (130.61 mg, 1.34 mmol, HCl) in DMF (10.00 mL) were added PyBOP (589.60 mg, 1.13 mmol), HOBt (153.09 mg, 1.13 mmol), and DIEA (798.70 mg, 6.18 mmol, 1.08 mL) at 20 °C. The mixture was stirred at 20 °C for 3 h. The mixture was diluted with 50 mL of water and extracted with EtOAc (30 mL x 4). The organic phases were washed with HCl (1 N, 30 mL), saturated NaHCO ₃ and brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 5/1 to 1/5) to give the title compound (290.00 mg, 688.04 μmol, yield 66.80%) as a colorless oil.

Step G. (3R)-tert-Butyl-8-acetyl-3,10-dimethyl-11-oxo-3,4,8,9,10, 11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-tert-butyl 8-(methoxy(methyl)carbamoyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (290.00 mg, 688.04 μmol) in THF (6.00 mL) was added MeMgBr (3 M, 1.15 mL) dropwise at -78 °C with stirring, then the mixture was stirred at 0 °C for 2 h. The mixture was poured into HCl (1N, 10 mL) at 0 °C, then extracted with EtOAc (20 mL×3), and the combined organic phases were successively washed with saturated NaHCO3 (20 ml) and brine (20 ml), then dried over _{anhydrous Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give the title compound ( _{260.00 mg, crude) as a white solid, which was used directly in the next step without purification. MS (ESI): mass calcd for Ci9H28N4O4 , 376.2} ; mass/charge found, 377.3 [M+ _H ]+.

Step H. (3R)-tert-Butyl-8-(3-(dimethylamino)acryloyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. (3R)-tert-Butyl-8-acetyl-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (260.00 mg, crude) was dissolved in NN-dimethylformamide dimethyl acetal (DMF-DMA) (8.00 mL), and the resulting mixture was heated to 70 °C with stirring for 16 h. The mixture was heated to 80 °C with stirring for 16 h, then concentrated under reduced pressure. The residue was dissolved in 20 mL of EtOAc. The resulting organic phase was washed with water (10 mL) and brine (20 mL), dried over anhydrous Na2SO4, filtered and the filtrate concentrated under reduced pressure to give the title compound (300.00 mg, crude) as a yellow oil, which was used directly in the next step without purification. MS (ESI): mass calcd for _{C22H33N5O4 , 431.2} ; mass/charge found, 432.2 [M+H] ₊ .

Step I. (3R)-tert-Butyl-3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-tert-butyl 8-(3-(dimethylamino)acryloyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (300.00 mg, crude) in EtOH (6.00 mL) was added NH2NH2-H2O (69.60 mg, 1.39 mmol, 67.57 μL), then the mixture was heated to 80 °C with stirring for 2 h. The mixture

- 156 047929 was concentrated under reduced pressure and the resulting residue was purified by preparative TLC (EtOAc/MeOH=20/1) to give the title compound (200.00 mg, 494.42 μmol, yield 71.12%, purity 99%) as a colorless oil.

Step J. (3R,8S*)-tert-Butyl-3,10-dimethyl-11-oxo-8-(III-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. (3R)-tert-Butyl 3,10-dimethyl-11oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine 2(7H)-carboxylate (200.00 mg, 499.41 μmol) was separated by SFC (column: AD (250 mm * 30 mm, 10 μm); mobile phase: [0.1% NH ₃ -H ₂ O IPA]; B%: 25% - 25%, 1.6 min; 50 min) to give the title compound (90.00 mg, 224.74 μmol, peak 1 on SFC (column AD-3S_3_5_40_3ML: Chiralpak AD-3 100x4.6 mm ID, 3 μm, mobile phase: methanol (0.05% DEA) in _CO2 from 5% to 40%, flow rate: 3 mL/min, wavelength: 220 nm), retention time: 2.178 min) as a white solid and (3R,8R*)-tert-butyl 3,10-dimethyl-11-oxo-8-(N-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-N-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (80.00 mg, 199.77 μmol, peak 2 on SFC, retention time: 2.328 min) as a white solid.

Intermediate 9a: (3R,8R*)-tert-butyl 3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

The title compound was separated by SFC from (3K)-tert-butyl-3,10-dimethyl-11oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (product from intermediate 9, step I) in a manner analogous to that for intermediate 7, step J: (80.00 mg, 199.77 μmol, peak 2 on SFC (column AD3S_3_5_40_3ML: Chiralpak AD-3 100x4.6 mm ID, 3 μm, mobile phase: methanol (0.05% DEA) in _CO2 from 5% to 40%; flow rate: 3 ml/min, wavelength: 220 nm), retention time: 2.328 min). MS (ESI): mass calcd. for C ₂₀ H ₂₇ N ₅ O ₄ , 401.2; mass/charge, found value, 402.3 [M+H]+.

Intermediate 10a: (3R,8S*)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

Step A. (3R)-2-(tert-Butoxycarbonyl)-3,10-dimethyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-Spyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-8-carboxylic acid. To a solution of (3R)-2-tert-butyl-8-ethyl-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2,8(7H)-dicarboxylate (intermediate 8, step D; 2.50 g, 6.15 mmol) in THF (25.00 mL) and water (5.00 mL) was added NaOH (369.00 mg, 9.23 mmol). The mixture was stirred at 40 °C for 16 h, then diluted with water (30 mL) and the pH was adjusted to 5 by adding HCl (1 N). The resulting mixture was extracted with ethyl acetate (50 mL x 3), the combined organic phases were washed with brine (40 mL), dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give the title compound (2.00 g, 5.29 mmol, 85.94% yield) as a white solid. MS (ESI): mass calcd for C ₁ gH ₂ 6N ₄ O ₅ , 378.2; mass/charge found, 379.2 [M+H] ⁺ .

Step B. (3R)-tert-Butyl 8-(methoxy(methyl)carbamoyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-2-(tertbutoxycarbonyl)-3,10-dimethyl-11-oxo-2,3,4,7,8,9,10,11-octahydro-N-pyrido-[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-8-carboxylic acid (2.00 g, 5.28 mmol) and N-methoxymethanamine (669.51 mg, 6.86 mmol, HCl) in DMF (30.00 mL) were added PyBOP (3.02 g, 5.81 mmol), HOBt (784.78 mg, 5.81 mmol), and DIEA (4.09 g, 31.68 mmol, 5.52 mL). The mixture was then stirred at 20°C for 3 h. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic layers were sequentially

- 157 047929 was washed thoroughly with water (100 mL x 2), 1N HCl (50 mL) and sat. aq. NaHCO3 (50 mL). Then the organic portion was dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/EtOAc=50% ~ 100%) to give the title compound (2.06 g, 4.89 mmol, yield 92.56%) as a white solid. MS (ESI): mass calcd for C ₂₀ H ₃₁ N ₅ O ₅ , 421.2; mass/charge found, 422.3 [M+H]+.

Step C. (3R)-tert-Butyl-8-acetyl-3,10-dimethyl-11-oxo-3,4,8,9,10, P-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-tert-butyl 8-(methoxy(methyl)carbamoyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-N-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxylate (2.06 g, 4.89 mmol) in THF (30 mL) was added dropwise MeMgBr (3 M, 4.89 mL) at -30 °C under N2. The mixture was warmed to 0 °C with stirring for 1 h. The mixture was slowly poured into 1 N HCl (100 mL) and extracted with ethyl acetate (80 mL×3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give the title compound (1.67 g, crude) as a white solid. MS (ESI): mass calcd for _{Ci9H28N4O4 , 376.2} ; mass/charge found, 377.1 [M ₊ H]+.

Step D. (3R)-tert-Butyl 8-((E)-3-(dimethylamino)acryloyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. (3R)-tert-Butyl-8-acetyl-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (400 mg, 1.06 mmol) was dissolved in DMF-DMA (1.79 g, 15.06 mmol, 2 mL). The mixture was stirred at 75 °C for 24 h. The mixture was extracted with ethyl acetate (40 mL x 2) and washed with water (20 mL). The combined organic layers were washed with water (30 mL x 2). _The organic portion was dried over anhydrous _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure to give the title compound (400 mg, 926.94 μmol, yield 87.24%) as a brown solid.

Step E. (3R)-tert-Butyl-8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of (3R)-tert-butyl 8-((E)-3(dimethylamino)acryloyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (200 mg, 463.47 μmol) in pyridine (5 mL) was added NH ₂ OH-HCl (193.24 mg, 2.78 mmol). The mixture was stirred at 115 °C for 16 h, then diluted with water (30 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with 1 N HCl. HCl (20 mL x 2), dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (EtOAc=1) to give (3R)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (87 mg, 166.87 μmol, yield 36.00%, purity 77%) as a brown oil. MS (ESI): mass calcd for C ₂₀ H ₂₇ N ₅ O ₄ , 401.2; mass/charge, value found, 402.3 [M+H]+.

Step F. (3R,8S*)-tert-Butyl-8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. (3R)-tert-Butyl 8-(isoxazol-3-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazene-2(7H)carboxylate (92 mg) was purified by HPLC (Column: Phenomenex Synergi C18 150*30 mm*4 μm; Mobile phase: [water (0.225% FA)-ACN]; B%: 33%-63%, 10.5 min) to give the title compound (60 mg), which was separated by SFC (Column: AD (250 mm*30 mm, 10 μm); Mobile phase: [0.1% NH3-H2O MEOH]; B%: 25% - 25%, 3.3 min; 70 min) to give (3R,8S*)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (peak 1 on SFC (column AD-3S_3_5_40_3ML: Chiralpak AD-3 100x4.6 mm ID, 3 μm, mobile phase: methanol (0.05% DEA) in CO2 from 5% to 40%, flow rate: 3 mL/min, wavelength: 220 nm), time retention time=1.662 min, 17 mg, 42.35 μmol, 18.48% yield) as a white solid and the diastereomer, (3R,8R*)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (peak 2, retention time=1.858 min, 17 mg, 42.35 μmol, 18.48% yield), as a white solid. MS (ESI): mass calcd for C ₂₀ H ₂₇ N ₅ O ₄ , 401.2; mass/charge, value found, 402.3 [M+H]+.

Intermediate IIa: (3R,8R*)-tert-butyl-8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

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L?

Mn

The title compound was separated by SFC from (3R)-tert-butyl 8-(isoxazol-3-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxylate (product from intermediate 4, step E) in a manner analogous to that for intermediate 10, step F. (17 mg, 42.35 μmol, peak 2 on SFC (column AD3S_3_5_40_3ML: Chiralpak AD-3 100x4.6 mm ID, 3 μm, mobile phase: methanol (0.05% DEA) in CO2 from 5% to 40%, flow rate: 3 ml/min, wavelength: 220 nm), retention time=1.858 min). MS (ESI): mass calcd. for C ₂₀ H ₂₇ N ₅ O ₄ , 401.2; mass/charge, found value, 402.3 [M+H]+.

Intermediate 12a: (3R,8S*)-tert-butyl-8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1 H-pyrido[4',3'3^pyrazolo^3-ya][1',4]diazepine-2(7H)-carboxylate /^N'+ .O /sX

NN γχ,χ Y J ⁰ ν Ν

Vos

Step A. (3R)-tert-Butyl 8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate.

To a solution of (3R)-tert-butyl 8-((E)-3-(dimethylamino)acryloyl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 10, step D; 200 mg, 463.47 μmol) in HOAc (3 mL) and water (6 mL) was added NH2OH-HC1 (193.24 mg, 2.78 mmol). The mixture was stirred at 10 °C for 16 h and then heating was continued for another 48 h at 70 °C. At this time, the mixture was concentrated under reduced pressure, and the resulting residue was diluted with THF (3 mL), and the pH was adjusted to 9~10 with aq. _{Na2CO3 . Boc2O} (303.46 mg, 1.39 mmol, 319.43 μL) was added and the mixture was stirred at 10 °C for 16 h. The mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL×3). _The combined organic layers were dried over _Na2SO4 , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (EtOAc/MeOH=10/1) to give a mixture (140 mg, 327.48 μmol, yield 70.66%, purity 93.8%) of the title compound, (3R)-tert-butyl 8-(isoxazol5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine2(7H)-carboxylate, and (3R)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate as a colourless oil. MS (ESI): mass calcd for C ₂₀ H ₂ 7N ₅ O ₄ , 401.2; m/z found, 402.3 [M+H] ⁺ .

Step B. (3R,8S*)-tert-Butyl-8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate.

A mixture of (3R)-tert-butyl 8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate and (3R)-tert-butyl 8-(isoxazol-3-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxylate (140 mg) was separated by SFC (column: AD (250 mm * 30 mm, 5 μm); mobile phase: [0.1% NH3-H2O IPA]; 2.6 min; 120 min) to give the title compound, (3R,8S*)-tert-butyl 8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (peak 1 on SFC (column AD-3S_5_5_40_3ML: Chiralpak AD-3 100x4.6 mm ID, 3 μm, mobile phase: ethanol (0.05% DEA) in CO2 from 5% to 40%, flow rate: 3 mL/min, wavelength: 220 nm), retention time: 1.753 min, 36 mg, 89.67 μmol, 27.41% yield) as a colourless oil and a mixture (peak 2 and peak 3, retention times: 1.866 and 1.907 min, 50 mg, containing intermediate 7). MS (ESI): mass calcd for C ₂₀ H ₂ 7N ₅ O ₄ , 401.2; mass/charge found, 402.3 [M+H] ⁺ .

Intermediate 13a: (3R,8R*)-tert-butyl-8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a] [1,4]diazepine-2(7H)-carboxylate

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The mixture (peak 2 and peak 3, retention time: 1.866 and 1.907 min, 50 mg) from the product of intermediate 12, step B was further purified by SFC (column: OJ (250 mm * 30 mm, 5 μm); mobile phase: [0.1% NH3-H2O IPA]; B%: 20%-20%, 2.2 min; 30 min) to give the title compound (peak 1 by SFC (column OJ-3S_4_5_40_3ML: Chiralcel OJ-3 100x4.6 mm ID, 3 μm, mobile phase: isopropanol (0.05% DEA) in CO2 from 5% to 40%, flow rate: 3 mL/min, wavelength: 220 nm), retention time=1.233 min, 11 mg, 27.40 μmol, yield 8.38%). MS(ESI): mass calcd. for C ₂₀ H ₂₇ N ₅ O ₄ , 401.2; observed mass/charge value 402.3 [M+H]+.

Intermediate 14a: tert-butyl 10-methyl-8-methylene-11-oxo-3,4,8,9,10,11-hexahydro1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

Method A.

Step A. 5-Methylene-1,3,2-dioxathiane 2-oxide. To a solution of 2-methylenepropane-1,3-diol (5.00 g, 56.75 mmol, 4.63 mL, 1.00 equiv) in CCl ₄ (50.00 mL) was added a solution of SOCl ₂ (10.13 g, 85.13 mmol, 6.18 mL, 1.50 equiv) in CCl ₄ (10.00 mL) at 0 °C under N 2 and the mixture was stirred at 0 °C for 45 min. The mixture was concentrated under reduced pressure to give 5-methylene-1,3,2-dioxathiane 2-oxide (6.90 g, 51.43 mmol, 90.63% yield) as a yellow oil, which was used directly in the next step. ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.36–5.39 (m, 2H), 5.16 (s, 2H), 4.22–4.28 (m, 2H).

Step B: 2-((Methylamino)methyl)prop-2-en-1-ol: A solution of 5-methylene-1,3,2-dioxathiane 2-oxide (1.00 g, 7.45 mmol, 1.00 equiv) and methanamine (2 M, 11.18 mL, 3.00 equiv) in THF (2.00 mL) was heated to 70 °C for 16 h. The mixture was filtered and the filtrate was concentrated in vacuo to give the title compound (750.00 mg, 7.41 mmol, 99.46% yield) as a yellow oil, which was used directly in the next step.

Step C. tert-Butyl (2-(hydroxymethyl)allyl)(methyl)carbamate. To a solution of 2-(methylaminomethyl)prop-2-en-1-ol (750.00 mg, 7.41 mmol, 1.00 equiv) in dioxane (5.00 mL)/ _H2O (5.00 mL) were added _Boc2O (1.94 g, 8.89 mmol, 2.04 mL, 1.20 equiv) and _NaHCO3 (622.40 mg, 7.41 mmol, 1.00 equiv). The mixture was stirred at 30 °C for 16 h. The mixture was diluted with EA (50 mL) and washed with brine (50 mL). _The organic phase was dried over _Na2SO4 , filtered and concentrated to give a yellow oil which was purified by silica gel column to give the title compound (710.00 mg, 3.53 mmol, 47.61% yield) as a yellow oil. 1H NMR (400 MHz, CD3OD) δ 5.10 (s, 1H), 4.97 (s, 1H), 3.91-4.10 (m, 4H), 2.81 (s, 3H), 1.50 (s, 9H).

Step D: 2-((methylamino)methyl)prop-2-en-1-ol hydrochloride: To a solution of tert-butyl N-[2(hydroxymethyl)allyl]-N-methylcarbamate (710.00 mg, 3.53 mmol, 1.00 equiv) in dioxane (3.00 mL) was added HCl/dioxane (4 M, 5.00 mL, 5.67 equiv) and the mixture was stirred at 15 °C for 1 h. The mixture was concentrated in vacuo to give the title compound (480.00 mg, 3.49 mmol, 98.81% yield, HCl) as a yellow oil, which was used directly in the next step. 1H NMR (400 MHz, CD3OD) δ 5.46 (s, 1H), 5.32 (s, 1H), 4.20 (s, 2H), 3.71 (s, 2H), 2.73 (s, 3H).

Step E. tert-Butyl 3-((2-(hydroxymethyl)allyl)(methyl)carbamoyl)-6,7-dihydro-2H-pyrazolo[4,3-pyridine-2-carboxylate. A mixture of 5-tert-butoxycarbonyl-1,4,6,7-tetrahydropyrazolo[4,3c]pyridine-3-carboxylic acid (550.00 mg, 2.06 mmol, 1.00 equiv), DIPEA (798.70 mg, 6.18 mmol, 1.08 mL, 3.00 equiv), HATU (939.93 mg, 2.47 mmol, 1.20 equiv), and 2-(methylaminomethyl)prop-2-en-1-ol (425.21 mg, 3.09 mmol, 1.50 equiv, HCl) in DMF (6.00 mL) was heated to 80 °C for 16 h. The mixture was diluted with EtOAc (80 mL) and washed with brine (80 mL * 3). The organic phase was dried over _Na2SO4 , filtered and concentrated under reduced pressure to give a yellow _oil . The yellow oil was purified by silica gel column to give the title compound (390.00 mg, 1.11 mmol, 54.03% yield) as a yellow solid. LCMS: 351 [M+1].

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Stage F. tert-Butyl 10-methyl-8-methylene-11 -oxo-3,4,8,9,10,11 -hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1 ,4]diazepine-2(7H)-carboxylate. To a solution of tert-butyl 3-[2-(hydroxymethyl)allylmethylcarbamoyl]-2,4,6,7-tetrahydropyrazolo[4,3-c]pyridine-5-carboxylate (200.00 mg, 570.76 μmol, 1.00 equiv) and triphenylphosphane (194.62 mg, 741.99 μmol, 1.30 equiv) in THF (3.00 mL) was added DIAD (150.04 mg, 741.99 μmol, 144.27 mL, 1.30 equiv) and the mixture was stirred at 30 °C for 4 h. The mixture was diluted with EtOAc (50 mL) and washed with HCl (1 M, 50 mL). The organic phase was dried over _Na2SO4 , filtered and concentrated in vacuo to give an oil. The oil was purified by silica _gel column to give the title compound as a crude product (320.00 mg, crude, containing _Ph3PO ) as a yellow oil. LCMS: 355 [M+23].

Method B.

Step A. 5-tert-Butyl 3-ethyl-2-(2-(chloromethyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine 3,5(4H)-dicarboxylate. To a solution of 3-chloro-2-(chloromethyl)prop-1-ene (7.62 g, 60.95 mmol, 7.05 mL, 3.00 equiv) in DMF (100.00 mL) were added 5-tert-butyl 3-ethyl-2,4,6,7-tetrahydropyrazolo[4,3-c]pyridine 3,5-dicarboxylate (6.00 g, 20.32 mmol, 1.00 equiv) and K2CO3 (3.65 g, 26.41 mmol, 1.30 equiv). The mixture was stirred at 25 °C for 6 h and then heated to 75 °C for 16 h. The mixture was diluted with EtOAc (80 mL), washed with HCl (1 M, 80 mL) and brine (80 mL * 2). The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a yellow oil. The oil was purified by silica gel column to give the title compound (2.90 g, 7.55 mmol, 37.18% yield) as a colorless oil.

Step B. tert-Butyl 10-methyl-8-methylene-11-oxo-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. A solution of 5-tert-butyl 3-ethyl-2-(2-(chloromethyl)allyl)6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-3,5(4H)-dicarboxylate (1.00 g, 2.61 mmol, 1.00 equiv) and methanamine (7.5 M, 40.00 mL, 33% purity, 114.94 equiv) in EtOH (30.00 mL) was heated to 80 °C in a sealed tube for 16 h. The mixture was concentrated in vacuo to give a yellow oil. The yellow oil was purified by silica gel column to give the title compound (560.00 mg, 1.68 mmol, 64.37% yield) as a yellow oil. LCMS: 333 [M+1].

Intermediate 15a: tert-butyl 8-hydroxy-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

Vis

Step A. tert-Butyl 10-methyl-8,11-dioxo-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of tert-butyl 10-methyl-8-methylene-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 14a, 1.4 g, 4.21 mmol) in THF (50 mL) were added _OsO4 (107.08 mg, 421.18 μmol, 21.85 μL) and a solution of _NaIO4 (2.70 g, 12.64 mmol, 700.16 μL) in H2O (25 mL) at 0 °C, and the mixture was stirred at 15 °C for 16 h. The reaction mixture was quenched with saturated aqueous _NaHCO3 (30 ml) at 0 °C, then diluted with water (50 ml) and extracted with ethyl acetate (50 ml x 2). The combined organic layers were washed with brine (50 ml), dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ ) to give the title compound (0.9 g, 2.69 mmol, 63.91% yield) as a yellow solid. MS (ESI): mass calcd for C ₁₆ H ₂₂ N ₄ O ₄ , 334.4; m/z found, 279 [M+H-56]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.01(s, 2H), 4.66(s, 2H), 4.04(s, 2H), 3.73(t, J=5.1 Hz, 2H), 3.20(s, 3H), 2.76(s, 2H), l.49(s, 9H).

Step B. tert-Butyl 8-hydroxy-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate. To a solution of phenylmagnesium bromide (3 M, 2.09 mL) in THF (2 mL) was added a solution of tert-butyl 10-methyl-8,11-dioxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (0.7 g, 2.09 mmol) in THF (10 mL) at 30 °C and the mixture was stirred at 15 °C for 2 h under N ₂ . The reaction mixture was quenched with saturated NH ₄ Cl solution (5 mL) and extracted with EtOAc (3 mL x 2). The combined organic layers were washed with brine (10 mL) and dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure.

The residue was purified by preparative TLC (SiO2) to give the title compound (0.320 g, 721.49 μmol, 34.46% yield, 93% purity) as a white solid. MS (ESI): mass calcd for C ₂₂ H ₂₈ N ₄ O ₄ , 412.5; mass/charge found, 413.1 [M+H]+. 1H NMR (400

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MHz, CDCl ₃ ) δ 7.60 (d, J=7.5 Hz, 2H), 7.47-7.31 (m, 3H), 4.90 (d, J=14.4 Hz, 1H) , 4.63 (s, 2H), 4.46 (d, J=14.4 Hz, 1H), 3.82 (d, J=5.5, 13.4 Hz, 1H), 3.65- 3.36 (m, 3H), 3.19 (s, 3H), 2.79-2.65 (m, 2H), 1.49 (s, 9H).

Intermediate 16a: tert-butyl 8-fluoro-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate

To a solution of tert-butyl 8-hydroxy-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro-N-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 15a, 0.08 g, 180.37 μmol) in DCM (1 mL) was added a solution of DAST (0.0625 g, 387.74 μmol, 51.23 μL) in DCM (1 mL) at 0 °C under N2. The mixture was stirred at 15 °C for 2 h. The reaction mixture was quenched with _H2O (3 mL) at 0 °C, then extracted with EtOAc (5 mL×2). The combined organic layers were washed with brine (5 mL), dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced _pressure . The residue was purified by preparative TLC ( _SiO2 ) to give the title compound (0.065 g, 156.83 μmol, 86.95 _{% yield) as a white solid. MS (ESI): mass calcd for C22H27N4O3F , 414.5.4} . 1H NMR (400 MHz, CDCl ₃ ) δ 7.517.33 (m, 5H), 4.90 (m, 1H), 4.73-4.52 (m, 3H), 3.98-3.83 (m, 1H), 3.77-3.48 (m, 3H), 3.23 (s, 3H), 2.80 (s, 2H), 1.54-1.39 (m, 9H).

Example 1 a. N-(3-Cyano-4-fluorophenyl)-10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-3,4,8,9,10,11hexahydro- 1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1 ^diazepine^rCH-carboxamide

Step A. 10-Methyl-8-(1H-1,2,4-triazol-3-yl)-3,4,7,8,9,10-hexahydro-1H-pyrido[4',3':3,4]-pyrazolo[1,5-a][1,4]diazepin-11(2H)-one.

To a solution of tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (Intermediate 2, 40.00 mg, 103.24 μmol) in DCM (3.00 mL) was added TFA (770.00 mg, 6.75 mmol, 500.00 μL). The reaction mixture was stirred at 15 °C for 30 min. The mixture was concentrated under reduced pressure to give the title compound (40.00 mg, crude, TFA salt) as a yellow oil, which was used directly in the next step without purification.

Step B. N-(3-Cyano-4-fluorophenyl)-10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-3,4,8,9,10 ,11hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H-carboxamide.

To a mixture of 10-methyl-8-(1H-1,2,4-triazol-3-yl)-2,3,4,7,8,9-hexahydro-1H-pyrido[2,3]pyrazolo[2,4B][1,4]diazepin-11-one (40.00 mg, 99.67 μmol, TFA) in DCM (2.00 mL) was added TEA (40.34 mg, 398.66 μmol, 55.26 μL), followed by phenyl N-(3-cyano-4-fluorophenyl)carbamate (25.54 mg, 99.67 μmol). The reaction mixture was stirred at 15 °C for 16 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified twice by RP-HPLC (condition A) to give the title compound (28.00 mg, 61.68 μmol, yield 61.88%, purity 99%) as a white solid. MS (ESI): mass calcd for C ₂₁ H ₂₀ FN ₉ O ₂ 449.2; m/z found 450 [M+H]+. 1H NMR (400 MHz, CDCl ₃ ) δ 8.25 (s, 1H), 7.78 (dd, J=2.8, 5.40 Hz, 1H), 7.56-7.64 (m, 1H), 7.13 (t, J=8.7 Hz, 1H), 6.88 (s, 1H), 4.75-4.85 (m, 2H), 4.65-4.74 (m, 2H), 3.90-4.02 (m, 2H), 3.76-3.87 (m, 3H), 3.15 (s, 3H), 2.87 (t, J=5.7 Hz, 2H).

Example 2a. 0-(3-Cyano-4-fluorophenyl)-10-methyl-8-(1,2,4-oxadiazol-5-yl)-11-oxo-3,4,8,9,10,11hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H-carboxamide

- 162 047929

The title compound was prepared in a manner analogous to Example 1a, substituting tert-butyl 10-methyl-8-(1,2,4-oxadiazol-5-yl)-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 3a) for tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calc. for C ₂₁ H ₁₉ FN ₈ O ₃ 450.2; mass/charge, found value, 451.2 [M+H]+. 1H NMR (400 MHz, DMSOd6) δ 9.02 (s, 1H), 8.43 (s, 1H), 7.93 (dd, J=2.8, 5.77 Hz, 1H), 7.79 (ddd, J=2.8, 4.9, 9.3 Hz, 1H), 7.42 (t, J=9.2 Hz, 1H ), 4.57 (d, J=3.1 Hz, 2H), 4.38-4.45 (m, 1H), 4.28-4.35 (m, 1H), 3.63-3.80 (m, 3H), 3.58 (br dd, J=5.4, 14.8 Hz, 2H), 3.02 (s, 3H), 2.91-3.01 (m , 2H).

Example 3a. N-(3-Cyano-4-fluorophenyl)-10-methyl-11-oxo-8-(1H-tetrazol-5-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Example 1a substituting tert-butyl 10-methyl-11-oxo-8-(1H-tetrazol-5-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 4a) for tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd. For C ₂₀ H ₁₉ FN ₁₀ O ₂ 450.2; mass/charge, found value, 451.2 [M+H]+. 1H NMR (400 MHz, DMSO-d ₆ ) δ 9.05 (s, 1H), 7.93 (dd, J=2.8, 5.8 Hz, 1H), 7.79 (ddd, J=2.8, 4.9, 9.3 Hz, 1H), 7.42 (t, J=9.2 Hz, 1H), 4.74 (dd, J =7.3, 14.3 Hz, 1H), 4.61 (s, 2H), 4.53 (dd, J=6.7, 14.2 Hz, 1H), 4.09-4.18 (m, 1H), 3.62-3.83 (m, 4H), 3.01 (s, 3H), 2.67-2.73 (m, 2H).

Example 4a. N-(3-Chloro-4-fluorophenyl)-10-methyl-11-oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Example 1a using tert-butyl 10-methyl-11-oxo-8-(III-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 5a) instead of tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4- 163 047929

[b][1,4]diazepine-2-carboxylate (intermediate 2a) in step A and phenyl (3-chloro-4-fluorophenyl)carbamate instead of phenyl (3-cyano-4-fluorophenyl)carbamate in step B. MS (ESI): mass calcd for C ₂₁ H ₂₁ N ₇ O ₂ ClF, 457.1; m/z found, 458.1 [M+H]+. 1H NMR (400 MHz, CDCl ₃ ) δ 7.62 (dd, J=2.6, 6.5 Hz, 1H), 7.56 (d, J=2.2 Hz, 1H), 7.28-7.22 (m, 1H), 7.10-7.03 (m, 2H), 6.28 (d, J=2.3 Hz, 1H), 4.89-4.80 (m, 1H), 4.75-4.64 (m, 3H), 4.10-3.99 (m, 1H), 3.96-3.87 (m, 1H), 3.79-3.769 (m, 2H), 3.57 (m, 1H), 3.09 (s, 3H), 3.08-3.07 (m, 1 H), 2.88 (br t, J=5.4 Hz, 2H).

Example 5a. N-(3-Cyano-4-fluorophenyl)-10-methyl-8-(1-methyl-III-pyrazol-3-yl)-11-oxo-3,4,8,9,10,11hexahydro-1 H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Example 1a, substituting tert-butyl 10-methyl-8-(1-methyl-1H-pyrazol-3-yl)-11-oxo-3,4,8,9,10,11-hexahydro-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 6a) for tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calc. for C ₂₃ H ₂₃ N ₈ O ₂ F, 462.4; mass/charge, found value, 463.1 [M+H] ⁺ ; 1H NMR (400 MHz, CDCl ₃ ) δ 7.80 (dd, J=2.8, 5.4 Hz, 1H), 7.60 (ddd, J=2.8, 4.6, 9.1 Hz, 1H), 7.35 (d, J=2.2 Hz, 1H), 7.14 (t, J=8.7 Hz, 1H), 6.8 7 (s, 1H), 6.17 (d, J=2.3 Hz, 1H), 5.32 (s, 1H), 4.78-4.55 (m, 4H), 3.99-3.91 (m, 1H), 3.90 (s, 3H), 3.88-3.79 (m, 2H), 3.75-3.68 (m, 1H), 3.65-3.57 (m, 1H), 3.17 (s, 3H), 2.88 (t, J=5.7 Hz, 2H).

Example 6a. N(3-Cyano-4-fluorophenyl)-10-methyl-8-(1-methyl-III-pyrazol-5-yl)-11-oxo-3,4,8,9,10,11hexahydro-1 H- pyrido[4',3':3,4]pyrazolo[1,5-a][1 ^diazepine A^N-carboxamide l F

The title compound was prepared in a manner analogous to Example 1a, substituting tert-butyl 10-methyl-8-(1-methyl-1H-pyrazol-5-yl)-11-oxo-3,4,8,9,10,11-hexahydro-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 7a) for tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calc. for C ₂₃ H ₂₃ N ₈ O ₂ F, 462.2; mass/charge, found value, 463.2 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) 5=7.79 (dd, J=2.8, 5.4 Hz, 1H), 7.61 (ddd, J=2.9, 4.6, 9.1 Hz, 1H), 7.49 (d, J=2.0 Hz, 1H), 7.15 (t, J=8.7 Hz, 1H), 6.80 (s, 1H), 6.25 (d, J=2.0 Hz, 1H), 4.73 (s, 2H), 4.67 (m, 1H), 4.51 (m, 1H), 3.94-3.79 (m, 6H), 3.67 (m, 1H), 3.44 (m, 1H), 3.16 (s, 3H), 84 (m, 2H).

Example 7a. (3R,8S*)-N-(3-Cyano-4-fluorophenyl)-3,10-dimethyl-11-oxo-8-(III-pyrazol-3-yl)3,4,8,9,10, 11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

- 164 047929

Step A. (3R,8S*)-3,10-Dimethyl-8-(1H-pyrazol-3-yl)-3,4,7,8,9,10-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11(2H)-one.

To a solution of tert-butyl (3R,8S*)-3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (Intermediate 7a, 90.00 mg, 224.74 μmol) in DCM (3.00 mL) was added TFA (462.00 mg, 4.05 mmol, 300.00 μL) at 15 °C with stirring for 1 h. The mixture was concentrated under reduced pressure to give the title compound (100.00 mg, crude, TFA) as a yellow oil, which was used directly in the next step.

Step B. (3R,8S*)-N-(3-Cyano-4-fluorophenyl)-3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide.

To a solution of (3R,8S*)-3,10-dimethyl-8-(1H-pyrazol-3-yl)-2,3,4,7,8,9-hexahydro-1H-pyrido[2,3]pyrazolo[2,4-b][1,4]diazepin-11-one (50.00 mg, 120.66 μmol TFA) and phenyl N-(3-cyano-4-fluorophenyl)carbamate (30.92 mg, 120.66 μmol) in DCM (3.00 mL) was added TEA (73.26 mg, 723.96 μmol, 100.36 μL) at 15 °C with stirring for 16 h. The mixture was concentrated under reduced pressure and the resulting residue was purified by RP-HPLC (method A) to give the title compound (24.00 mg, 51.89 µmol, 43.01% yield) as a white solid. MS (ESI): mass calcd for C ₂₃ H ₂₃ FN ₈ O2, 462.2; mass/charge found, 463.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.80 (dd, J=2.8, 5.4 Hz, 1H), 7.63-7.55 (m, 2H), 7.14 (t, J=8.7 Hz, 1H), 6.77 (s, 1H), 6.27 (d, J=2.4 Hz, 1H), 5.20- 5.09 (m, 1H), 4.84 (d, J=15.3 Hz, 1H), 4.75-4.60 (m, 2H), 4.51 (d, J=15.2 Hz, 1H), 3.88 (m, 1H), 3.79-3.71 (m, 1H), 3.67-3.59 (m, 1H), 3.15 (s 3H), 3.04 (dd, J=6.0, 15.9 Hz, 1H), 2.71 (d, J=15.9 Hz, 1H), 1.20 (d, J=6.90 Hz, 3H).

Example 8a. (3R,8S*)-N-(4-Fluoro-3-(trifluoromethyl)phenyl)-3,10-dimethyl-11-oxo-8-(1H-pyrazol-3yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Step B of Example 7a substituting phenyl (4-fluoro-3-(trifluoromethyl)phenyl)carbamate for phenyl (3-cyano-4-fluorophenyl)carbamate. MS (ESI): mass calcd for C ₂₃ H ₂₃ FN ₈ O ₂ , 505.2; m/z found, 506.2 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) δ 7.70 (dd, J=2.7, 6.1 Hz, 1H), 7.63-7.56 (m, 2H), 7.13 (t, J=9.5 Hz, 1H), 6.68 (s, 1H), 6.27 (d, J=2.4 Hz, 1H), 5.22- 5.11 (m, 1H), 4.84 (d, J=15.4 Hz, 1H), 4.75-4.59 (m, 2H), 4.52 (d, J=15.3 Hz, 1H), 3.93-3.83 (m, 1H), 3.79-3.71 (m, 1H), 3.67-3.58 (m, 1H), 3. 15 (s, 3H), 3.09-3.01 (m, 1H), 2.71 (d, J=16.1 Hz, 1H), 1.20 (d, J=6.9 Hz, 3H).

Example 9a. (3R,8R*)-N-(3-Cyano-4-fluorophenyl)-3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

- 165 047929

The title compound was prepared in a manner analogous to Example 7a but using (3R,8S)-tert-butyl 3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-2(7H)-carboxylate (intermediate 9a) instead of (3R,8R)-3,10-dimethyl-8-(1H-pyrazol-3-yl)-3,4,7,8,9,10-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11(2H)-one (intermediate 8a). MS (ESI): mass calcd for C23H23FN8O2, 462.19; m/z found, 463.1 [M+H]+. ¹ H NMR (400 MHz, CDCl3) δ 7.80 (dd, J=2.76, 5.52 Hz, 1H), 7.54-7.64 (m, 2H), 7.13 (t, J=8.7 Hz, 1H), 6.80 (s, 1H), 6.27 (d, J=2.38 Hz, 1H), 5.15 (m, 1H), 4.86 (d, J=15.2 Hz, 1H), 4.61-4.75 (m, 2H), 4.49 (d, J=15.4 Hz, 1H), 3.90 (m, 1H), 3.59-3.76 (m, 2H), 3.16 (s, 3H), 3.05 (dd, J=5.7, 15 ,9 Hz, 1H), 2.69 (d, J=15.9 Hz, 1H), 1.20 (d, J=6.9 Hz, 3H).

Example 10a. (3R,8R*)-N-(4-Fluoro-3-(Ίfluoromethyl)phenyl)-3,10-dimethyl-11-oxo-8-(III-pyrazol-3yl)-3,4,8,9, 10,11-hexahydro-1H-πpyrido[4',3':3,4]πyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Example 7a, substituting (3R,8R*)-tert-butyl 3,10-dimethyl-11-oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-2(7H)-carboxylate (intermediate 9a) for (3R,8S*)-3,10-dimethyl-8-(1H-pyrazol-3-yl)-3,4,7,8,9,10-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepin-11(2H)-one (intermediate 8) in step A and using phenyl (4-fluoro-3-(trifluoromethyl)-phenyl)carbamate instead of phenyl (3-cyano-4-fluorophenyl)carbamate in step B. MS (ESI): mass calcd for C ₂₃ H ₂ 3F ₄ N ₇ O ₂ , 505.2; m/z found, 506.1 [M+H] ⁺ . 1H NMR (400 MHz, CDCls) δ 7.70 (dd, J=2.64, 6.02 Hz, 1H), 7.63-7.56 (m, 2H), 7.13 (t, J=9.4 Hz, 1H), 6.73 (s, 1H), 6.27 (d, J=2.3 Hz, 1H), 5.16 (m , 1H), 4.86 (d, J=15.18 Hz, 1H), 4.73-4.61 (m, 2H), 4.50 (d, J=15.3 Hz, 1H), 3.90 (m, J=6.4 Hz, 1H), 3.75-3.60 (m, 2H), 3.17 (s, 3H), (m, 1H), 2.69 (d, J=15.9 Hz, 1H), 1.20 (d, J=6.9 Hz, 3H).

Example 11a. (3R,8S*)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

- 166 047929

The title compound was prepared in a manner analogous to Example 1a but using (3R,8S*)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 10a) instead of tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd for C ₂₃ H ₂₂ FN ₇ O ₃ , 463.2; mass/charge found, 464.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.48 (d, J=1.5 Hz, 1H), 7.83-7.77 (m, 1H), 7.61-7.53 (m, 1H), 7.20-7.11 (m, 1H), 6.58 (s, 1H), 6.36 (d, J=1.6 Hz, 1H), 5.24-5.06 (m, 1H), 4.82 (d, J=15.0 Hz, 1H), 4.78-4.71 (m, 1H), 4.60-4.47 (m, 2H), 3.99-3.89 (m, 1H), 3.77 (d, J=5.4 Hz, 1H), 3.70 (d, J=5, 9 Hz, 1H), 3.19 (s, 3H), 3.08-2.99 (m, 1H), 2.71 (d, J=16.0 Hz, 1H), 1.241.14 (m, 3H).

Example 12a. (3R,8R*)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamid

The title compound was prepared in a manner analogous to Example 1a but using (3R,8R*)-tert-butyl 8-(isoxazol-3-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 11a) instead of tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd for C ₂₃ H ₂₂ FN ₇ O ₃ , 463.2; m/z found, 464.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.47 (d, J=1.5 Hz, 1H), 7.81 (dd, J=2.8, 5.44 Hz, 1H), 7.58 (ddd, J=2.8, 4.5, 9.1 Hz, 1H), 7.16 (t, J=8.7 Hz, 1H),

6.64 (s, 1H), 6.38 (d, J=1.6 Hz, 1H), 5.16 (m, J=6.11 Hz, 1H), 4.85 (d, J=15 ,6 Hz, 1H), 4.75 (dd, J=7.0, 14.37 Hz,

1H), 4.59 (dd, J=5.6, 14.43 Hz, 1H), 4.51 (d, 7=15.4 Hz, 1H), 3.99 (m, J=6.4 Hz, 1H), 3.80-3.65 (m, 2H), 3.22 (s,

3H), 3.07 (dd, J=5.9, 15.8 Hz, 1H), 2.71 (d, J=16.0 Hz, 1H), 1.22 (d, J=6.8 Hz, 3H).

Example 13a. (3R,8S*)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo

3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamig

The title compound was prepared in a manner similar to that of Example 1a, with

- 167 047929 using (3R,8S*)-tert-butyl 8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 12a) instead of tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd. for C ₂₃ H ₂₂ FN ₇ O ₃ , 463.2; mass/charge, found value, 464.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.30 (d, J=1.7 Hz, 1H), 7.80 (dd, J=2.7, 5.32 Hz, 1H), 7.61-7.55 (m, 1H), 7.16 (t, 7=8.7 Hz, 1H), 6.59 (s, 1H), 6.2 9 (d, J=1.2 Hz, 1H), 5.19-5.10 (m, 1H), 4.83 (d, J=15.4 Hz, 1H), 4.76 (dd, J=7.3, 14.1 Hz, 1H), 4.65-4.56 (m, 1H), 4.51 (d, J=15.2 Hz, 1H), 4.01 ( m, J=5.5 Hz, 1H), 3.86 (dd, J=5.3, 14.98 Hz, 1H), 3.64 (dd, J=5.6, 14.98 Hz, 1H), 3.17 (s, 3H), 3.05 (dd, 7=5.7, 15.96 Hz, 1H), 2.72 (d, J=16.5 Hz, 1H), 1.22 (d, J=7.0 Hz, 3H).

Example 14a. (3R,8R*)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxamide

The title compound was prepared in a manner analogous to Example 1a using (3R,8R*)-tert-butyl 8-(isoxazol-5-yl)-3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1H-nirido[4',3':3,4]nirazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate I3a) instead of tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd for C ₂₃ H ₂₂ FN ₇ O ₃ , 463.2; mass/charge found, 464.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.28 (d, J=1.6 Hz, 1H), 7.80 (dd, J=2.7, 5.3 Hz, 1H), 7.58 (m, 1H), 7.16 (t, J=8.7 Hz, 1H), 6.63 (s, 1H), 6.29 (d, J=1 ,22 Hz, 1H), 5.15 (m, J=6.97 Hz, 1H), 4.85 (d, J=15.3 Hz, 1H), 4.79-4.70 (m, 1H), 4.66-4.58 (m, 1H), 4.49 (d, J=15.3 Hz, 1H), 4.07-3.98 (m, 1H), 3.83 (dd, 7=5.7, 15.1 Hz, 1H), 3.68-3.58 (m, 1H), 3.19 (s, 3H), 3.06 (dd, J=5.8, 16.0 Hz, 1H), 2.70 (d, 7=15.4 Hz, 1H), 1.22 (d, J=7.0 Hz, 3H).

Example 15a. X(3-Cyano-4-fluorophenyl)-11-hydroxy-13-methyl-14-oxo-11-phenyl-4,8,9,13-tetrazatricyclo[7.5.0.0 ^2,7 ]tetradeca-1,7-diene-4-carboxamide

The title compound was prepared in a manner analogous to Example 1a substituting tert-butyl 8-hydroxy-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro-1H-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 15a) for tert-butyl 10-methyl-11-oxo-8-(1H-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4-c-diazepine-2(7H)-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd. for C ₂₅ H ₂₃ N ₆ O ₃ F, 474.2; mass/charge, found value, 475.2 [M+H]+. 1H NMR (400 MHz, CDCl ₃ ) δ 7.78 7.76 (m, 1H), 7.65-7.53 (m, 3H), 7.48-7.35 (m, 3H), 7.14 (t, J=8.7 Hz, 1H), 6.68 (s, 1H), 4.96 (d, J=14.3 Hz, 1H), 4.81-4.62 (m, 2H), 4.49 (d, J=14.4 Hz, 1H), 3.97-3.81 (m, 2H), 3.63-3.39 (m, 2H), 3.24 (s, 3H), 2.90 (t, J=5.7 Hz, 2H), 2.49 (s, 1H).

Example 16a. X(3-Cyano-4-fluorophenyl)-8-fluoro-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro1H-nirido[4',3':3,4]pyrazolo[1,5-a][1,4]diazene-2(7H)-carboxamide

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The title compound was prepared in a manner analogous to Example 1a substituting tert-butyl 8-fluoro-10-methyl-11-oxo-8-phenyl-3,4,8,9,10,11-hexahydro-III-pyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)-carboxylate (intermediate 16a) for tert-butyl 10-methyl-11-oxo-8-(III-1,2,4-triazol-3-yl)-1,3,4,7,8,9-hexahydropyrido[2,3]pyrazolo[2,4b][1,4]diazepine-2-carboxylate (intermediate 2a) in Step A. MS (ESI): mass calcd. for C25H22F2N6O2, 476.5; mass/charge, found value, 477.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.78 (d, J=2.8, 5.5 Hz, 1H), 7.60 (m, 1H), 7.50-7.34 (m, 5H), 7.14 (t, J=8.7 Hz, 1H), 6.81 (s, 1H), 4.97 (d, J=6.8, 14.7 Hz, 1H), 4.85-4.60 (m, 3H), 3.95-3.84 (m, 2H), 3.78-3.53 (m, 2H), 3.26 (s, 3H), 2.92 (t, J=5.8 Hz, 2H).

4. Activity of compounds of formula (I) against HBV.

Procedure.

Anti-HBV activity was measured using the HepG2.117 cell line, a stable, inducibly HBV-producing cell line that replicates HBV in the absence of doxycycline (Tet-off system). The HepG2 cell line is available from ATCC® under accession number HB-8065. Transfection of the HepG2 cell line can be as described in Sun and Nassal 2006 Journal of Hepatology 45 (2006) 636–645 Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus.

To analyze antiviral activity, HBV replication was induced, then treated with serially diluted compound in 96-well plates. After 3 days of treatment, antiviral activity was determined by quantifying intracellular HBV DNA using real-time PCR and an HBV-specific primer and probe set.

The cytotoxicity of the compounds was tested using HepG2 or HepG2.117 cells incubated for 3 days in the presence of the compounds. Cell viability was assessed using the PERKIN ELMER ATPlite Luminescence Assay System.

Results.

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Table 8

Connection number HBV-AVE-HepG2.117 ec ₅₀ (μM, average value) TOX-HepG2.117 cc ₅₀ (μM, average value) 1 9,628 > 50 2 1,146 > 50 3 0,188 25,26 4 0,091 17.94 5 0.213 > 50 6 0.287 27.98 7 0.399 > 50 8 <0.039 10.82 9 0,054 16.36 10 0,062 7.26 11 0,054 8.43 12 0,066 25,28 13 0,055 N.D. 14 0,085 6.96 15 0,055 > 50 16 0,079 > 50 17 0.436 > 50 18 0.252 > 50 19 0,079 > 50 20 0,108 > 50 21 0.268 > 50 22 0.129 > 50 23 0,022 > 50 24 0,036 10.41 25 0,016 >25 26 0,028 26,23 27 0.103 > 50 28 0,057 16.69 29 0,741 > 50 30 1,504 > 50 31 0,075 > 50 32 0.332 > 50 33 9,612 > 50 34 1,381 > 50 36 0,077 24.55 37 0.345 > 50 38 1,747 > 50

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39 2,487 > 50 40 0,022 20.93 41 0.328 > 50 42 0.308 1.44 43 0,210 > 50 44 0,079 > 50 45 0,192 > 50 46 0,505 > 50 47 0.249 3.63 48 1,228 > 50 C49 0.274 20.55 C50 H. d. C51 1,025 39.03 C52 0.995 > 50 C53 1,081 20,41 C54 1,994 13.61 C55*R >2.5 3.22 C55*S 0,069 > 50 C56 0.962 28.4 C57 0,04 18.85 C58 0.531 18 C59 0,576 > 50 C60 0,565 > 50 C61*S 0.153 > 50 C62 0.13 9.31 C63 0.824 8.21 C64 0,089 >49 C65 0.299 > 50 C61*R >2.5 8,H C66 0,042 9.18 C67 0,07 22.53 C68 0.321 10.55 C69 0,063 14.5 C70 0.232 > 50 C71 >2.5 > 50 C72 >2.5 > 50 C73 >2.5 > 50 C74 >2.5 > 50 C75 1,428 > 50 C76 0.983 >25 C77 0,038 > 50 C78 0,707 > 50 C79 > 1.6 >50 C80 0,05 26.11 C81 0,058 46.22 C82 0,019 32,52 C83 0.229 > 50 C84 0,311 9.16 C85*R 0,088 > 50 C85*S 0,035 > 50

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C86 0,046 >50 C87 0,066 > 50 C88 0,038 >48 C89 0,067 22.81 C90 0,667 21.94 C91 0,083 36.74 C92 0,025 24.45 C93 0,055 30 C94 0,162 4.77 C95 0,2 >47 C96 0,025 10.65 C97 0,011 9.79 C98 0,073 9.83 C99 0,072 > 50 C100 0,085 15,5 C101 0,036 8.95 C102 0,019 7.14 C103 0.39 >50 C104 0,632 > 50 C105 C105*R 0,059 14.56 C105*S 0.297 > 50 C106 0,031 33.79 C107 0,015 13.22 C108 0,085 20,02 C109 0,013 18.91 CHO 0,017 16.03 CHI 0,086 > 50 C112 0,025 > 50 C113 0.257 >50 C114 0,048 30.85 C115 0.265 > 50 C116 0,015 12.36 C117 0,119 18,17 C118 0,057 24.1 C119*R 0.269 > 50 C119*S 0,757 >50 C120 0,047 C121 >2.5 >50 C122 0.214 > 50 C123*R 0,174 19.98 C123*S 0,517 19,21 C124 0,011 26.7 C125 0,037 > 50 C88*R 0,076 > 50 C88*S 0,023 >50 C126 0,024 > 50 C127 0,133 15.73 C128 0,081 > 50

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C129 0,143 > 50 C130 0,082 33.66 C131 0,038 19.57 C132 0,061 >50 C133 0.332 > 50 C134 >2.5 > 50 C135 0,1 > 50 C136 >2.5 > 50 C137 0,056 > 50 C138 0,061 > 50 C139 0,04 > 50 C140 0,032 C141 0,056 > 50 C142 0.268 > 50 C143 0.16 > 50 C144 >2.5 > 50 C145 0.106 > 50 C146 1,794 > 50 C147 0,054 > 50 C148 0.385 > 50 C149 0,059 > 50 C150 0,075 > 50 C151 0.42 > 50 C152 0,145 > 50 C153 0,082 > 50 C154 0,071 9.53 C155 0,083 > 50 C156 0,174 > 50 C157 0,02 > 50 C158 2,381 > 50 C159 0.979 36.03 C160 0,029 > 50 C161 0,051 > 50 C162 0,082 >50 C163 0,098 > 50 C164 0,039 4.29 C165 0,676 > 50 C166 0,034 29.1 C167 0.277 > 50 C168 0,046 6.11 C169 0,831 > 50 C170 0,432 > 50 C171 0,082 15.33 C172 0.19 > 50 C173 0.41 > 50 C174 0.18 15.34 C175 0,038 35.5 C176 0.274 14.99 C177 0,136 > 50 C178 0.207 > 50

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C179 0.219 > 50 C180 0,123 > 50 C181 0,152 > 50 C182 0,075 > 50 C183 0,108 > 50 C184 >2.5 > 50 C185 0,114 > 50 C186 0,049 21.64 C187 0,155 > 50 C188 0,04 49.63 C189 0,043 41,81 C190 0,115 33,14 C191 0,097 32.95 C192 0,076 39.72 C193 0,197 > 50 C194 0,075 26.44 C195 0,566 > 50 C196 0,048 38.03 C197 0.24 > 50 C198 0,053 > 50 C199 0.33 > 50 C200 0.25 >50 C201 0.229 > 50 C202 0,071 > 50 C203 0.09 > 50 C204 0,028 19.2 C205 0,04 -1 C206 0,038 -1 C207 0,053 25,55 C208 0.13 > 50 C209 >2.5 > 50 C210 0,062 > 50 C211 0.285 > 50 C212 0,05 > 50 C213 0.382 37,52 C214 0,074 > 50 C215 0.106 > 50 C216 0.09 > 50 C217 0.183 39.54 C218 0,039 > 50 C219 0,068 > 50 C220 0,077 > 50 C221 0,022 > 50 C222 0,124 >50 C223 0.219 > 50 C224 0.326 >50 C225 0.263 > 50 C226 0.194 > 50 C227 0.129 37,33 C228 0,062 37,51

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C229 0.438 >25 C230 0,045 >12.5 C231 0.09 > 50 C232 0,066 24,44 C233 0.423 29.76 C234 0,036 >50 C235 1,934 > 50 C236 0.325 >51 C237 0,107 18.79 C238 0,029 39.9 C239 0.154 > 50 C240 0,017 35.23 C241 0.19 >50 C242 0,589 > 50 C243 >2.5 > 50 C244 0,048 24.52 C245 0,072 > 50 C246 0,171 > 50 C247 >2.5 > 50 C248 0,013 24.97 C249 >2.5 > 50 C250 0,033 >12.5

HepG2.117 cells with or without induction of HBc inclusions were cultured in the presence of DMSO or test compound in the absence of doxycycline. After fixation with formaldehyde and permeabilization with Triton-X-100, hepatitis B virus (HBc) core protein was labeled by immunochemical staining with primary antibodies to HBc. A secondary antibody conjugated to ALEXA 488 was used to detect primary HBV core signals by fluorescence. CELLMASK dark red and HOECHST 33258 were used to detect the cytoplasm and nucleus, respectively, to ensure separation of cellular compartments.

Image analysis software was used that allows detection of different morphological phenotypes, with determination of the level of HBV nuclei in the cytoplasm or nucleus (highly informative image analysis).

HBV replication inhibition assay.

Inhibition of HBV replication by the disclosed compounds was determined in cells infected or transfected with HBV or in cells with stably integrated HBV, such as HepG2.2.15 cells (Sells et al. 1987). In this example, HepG2.2.15 cells were maintained in cell culture medium containing 10% fetal bovine serum (FBS), geneticin, L-glutamine, penicillin, and streptomycin. HepG2.2.15 cells were seeded in 96-well plates at a density of 40,000 cells/well and treated with serially diluted compounds at a final concentration of 0.5% in DMSO, either alone or in combination by checkerboard addition of the drugs. Cells were incubated with compounds for 3 days, after which the medium was removed and fresh medium containing compounds was added to the cells and incubated for an additional 3 days. On day 6, the supernatant was removed and treated with DNase at 37°C for 60 min, followed by enzyme inactivation at 75°C for 15 min. Encapsidated HBV DNA was released from virions and covalently linked HBV polymerase by incubation in lysis buffer (Affymetrix QS0010) containing 2.5 μg proteinase K at 50°C for 40 min. HBV DNA was denatured by the addition of 0.2 M NaOH and detected using the QuantiGene branched DNA assay (BDNA) kit according to the manufacturer's recommendations (Affymetrix). HBV DNA levels were also quantified using qPCR based on amplification of encapsidated HBV DNA extracted with QuickExtraction solution (Epicentre Biotechnologies) and amplification of HBV DNA using HBV-specific PCR probes that are capable of hybridizing to HBV DNA and a fluorescently labeled probe for quantification. In addition, the viability of HepG2.2.15 cells incubated with test compounds alone or in combination was determined using the CellTitre-Glo reagent according to the manufacturer's protocol (Promega). The average background signal in wells containing culture medium alone was subtracted from all other samples, and the percent inhibition at each compound concentration was calculated by normalization to signals from HepG2.2.15 cells treated with 0.5% DMSO using equation E1.

E1: % inhibition = (DMSOavg - Xi)/DMSOcped x 100% where DMSOavg is the average signal calculated from the wells that were treated with DMSO control (0% inhibition for control) and Xi is the signal measured in individual wells. EC ₅₀ values, the effective concentrations at which inhibition is achieved, are

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50% inhibitory effect was determined by nonlinear fitting using Graphpad Prism software (San Diego, CA) and equation E2.

where Y represents the percent inhibition values and X represents the logarithm of the compound concentrations.

Selected disclosed compounds were analyzed using the HBV replication assay (BDNA assay) as described above, and an illustrative group of such active compounds is shown in Table 3. Table 9 shows the EC ₅₀ values obtained using the BDNA assay for a group of selected compounds.

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Table 9

Example number Name of the connection EC ₅ o for HBV DNA in HepG2.2.15 (µM) la N-(3-Cyano-4-fluorophenyl)-10-methyl-11-oxo-8-(1H-1,2,4- >4000 triazol-3-yl)-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2( 7H)carboxamide 2a T\G-(3-Cyano-4-fluorophenyl)-10-methyl-8-(1,2,4-oxadiazol5-yl)-11-oxo-3,4,8,9,10,11-hexahydro- 1Hpyrido[4',3': 3,4]pyrazolo[1,5-a] [1,4]diazepine-2(7H)carboxamide >4000 For N-(3 -Cyano-4-fluorophenyl)-10-methyl-11-oxo-8 -(1Htetrazol-5-yl)-3,4,8,9,10,11 -hexahydro-1Hpyrido[4',3 ': 3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide >4000 4a N-(3-Chloro-4-fluorophenyl)-10-methyl-11-oxo-8-(1Hpyrazol-3-yl)-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3 ': 3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 93 5a T4-(3-Cyano-4-fluorophenyl)-10-methyl-8-(1-methyl-1Hpyrazol-3-yl)-11-oxo-3,4,8,9,10,11-hexahydro-111pyr ido [4',3': 3,4]pyrazolo[1,5-a] [1,4]diazepine-2(7H)carboxamide 160 6a T4-(3-Cyano-4-fluorophenyl)-10-methyl-8-(1-methyl-1Hpyrazol-5-yl)-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[ 4',3': 3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 83 7a (3R, 8 S *)-N-(3-Cyano-4-fluorophenyl)-3,10-dimethyl-11oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10, 11-hexahydro-1Hpyrido[4',3': 3,4]pyrazolo[1,5-a] [1,4] diazepine-2(7H)carboxamide 37 8a (3R, 8 S * )-Ν-(4-Φτορ-3 -(trifluoromethyl)phenyl)-3,10-dimethyl-11 -oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11hexahydro- 1H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxamide 20 9a (3R, 8R*)-N-(3-Cyano-4-fluorophenyl)-3,10-dimethyl-11oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11 -hexahydro-1Hpyrido[4',3': 3,4]pyrazolo[1,5-a] [1,4] diazepine-2(7H)carboxamide 260 10a (3R, 8R*)-N-(4-OTop-3-(trifluoromethyl)phenyl)-3,10dimethyl-11-oxo-8-(1H-pyrazol-3-yl)-3,4,8,9,10,11hexahydro- 1H-pyrido[4',3':3,4]pyrazolo[1,5a][1,4]diazepine-2(7H)-carboxamide 280 On (3R, 8 S*)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-3-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 27 12a (ZK,8K*)-X-(3-Cyano-4-fluorophenyl)-8-(isoxazol-3-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11 - hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 240 13a (3R, 8 S *)-N-(3-Cyano-4-fluorophenyl)-8-(isoxazol-5-yl)3,10-dimethyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 22 14a (ЗК,8К*)-Х-(3-Cyano-4-fluorophenyl)-8-(isoxazol-5-yl)- 150

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3,10-d imethyl-11-oxo-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3':3,4]pyrazolo[1,5-a] [1,4] diazepine-2(7H)carboxamide 15a N-(3-Cyano-4-fluorophenyl)-11-hydroxy-13-methyl-14oxo-11-phenyl-4,8,9,13- tetrazatricyclo[7.5.0.02.7]tetradeca-1,7-diene-4carboxamide 140 16a N-(3-Cyano-4-fluorophenyl)-8-fluoro-10-methyl-11-oxo-8phenyl-3,4,8,9,10,11-hexahydro-1Hpyrido[4',3': 3, 4]pyrazolo[1,5-a][1,4]diazepine-2(7H)carboxamide 176

The disclosed subject matter is not intended to be limited in scope to the specific embodiments and examples described herein. Indeed, various modifications of the present invention in addition to those described will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to be within the scope of the appended claims.

All references (e.g., publications or patents or patent applications) cited herein are herein incorporated by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) were specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the scope of the following claims.

Claims (19)

1. Compound of formula (I) or its stereoisomer, where R ¹ is a 5-10 membered monocyclic or bicyclic ring, wherein the 5-10 membered monocyclic or bicyclic ring optionally contains 1-3 heteroatoms, wherein each of the heteroatoms is independently selected from N, O and S; and/or is optionally substituted with one or more substituents, each of which is independently selected from halogens, CN, CF3, CF2H, CFH2, CF2CH3, C alkyl, OC _1-6 alkyl, OCF3, OCF2H and C _3-4 cycloalkyl; ^R2 is selected from the group consisting of H, N-alkyl, and N-alkyl substituted with one or more F; n is an integer 0 or 1; W represents CHR ³ or C=CH ₂ ; R ³ is selected from the group consisting of H, F, OH, alkyl optionally substituted with OH or F, CONHC _1-4 alkyl, CONHC _3-6 cycloalkyl, NHSO ₂ C _1-4 alkyl and NHSO ₂ C _3-6 cycloalkyl; X is selected from the group consisting of CH2 and O; Y represents NR ⁵ ; R is selected from the group consisting of H, Ch^alkyl, Ch^alkyl substituted with one or more F, alkyl substituted with a 5- to 10-membered monocyclic or bicyclic aromatic ring, wherein the 5- to 10-membered monocyclic or bicyclic aromatic ring optionally contains one, two, three or four heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5- to 10-membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents each independently selected from the group consisting of halogens, CN, _CF3 , _CF2H , _CFH2 , _CF2CH3 , CH2CF3, alkyl, _OH , _OC1-4alkyl , OCF3, OCF2H, SO2N( _CH3 )2 and alkyl; and Z is selected from the group consisting of C(=O) and C(=S), - 178 047929 or a pharmaceutically acceptable salt thereof, provided that the compound is not 2. The compound according to claim 1, wherein R ¹ is a 5-9-membered monocyclic or bicyclic ring. 3. The compound of claim 1, wherein R ¹ is phenyl substituted with one or more substituents each independently selected from the group consisting of Cl, F, CN, CH ₃ , CF ₃ and cf ₂ h. 4. A compound according to claim 1, wherein R ¹ is phenyl substituted by one or more substituents representing chlorine, more particularly wherein R ¹ is dichlorophenyl. 5. The compound according to claim 1, wherein R ¹ is dichlorophenyl. 6. A compound according to any one of claims 1 to 5, wherein R ² is H or methyl. 7. A compound according to any one of claims 1 to 6, wherein Z is C=O. 8. A compound according to any one of claims 1 to 7, wherein X is CH ₂ . 9. A compound according to any one of claims 1 to 8, wherein W is _CH2 or CHF. 10. A compound according to any one of claims 1 to 9, wherein R ⁵ is selected from the group consisting of C _1.4 alkyl and C _1.4 alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are independently selected from N, O and S, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, CF ₃ , CF ₂ H, CFH ₂ , CF ₂ CH ₃ , C _1.4 alkyl, O-alkyl, OCF ₃ , OCF2H and O-cycloalkyl. 11. A compound according to any one of claims 1 to 9, wherein R ⁵ is O-alkyl substituted with a 5-10 membered monocyclic or bicyclic aromatic ring, wherein the 5-10 membered monocyclic or bicyclic aromatic ring optionally contains one, two or three heteroatoms, wherein the heteroatoms are N, and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, _{CF3 , CF2H} , CFH2, _{CF2CH3 , C1.4alkyl} , O-alkyl, _OCF3 , OCF2H and O-cycloalkyl. 12. The compound of claim 11, wherein the 5-10 membered monocyclic or bicyclic aromatic ring is phenyl, pyridyl, pyrazolyl or indolyl and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, CN, _CF3 , _CF2H , CFH2, _{CF2CH3 , C1.4alkyl} , _{OC1.4alkyl , OCF3 ,} OCF2H and O-cycloalkyl. 13. The compound of claim 12, wherein the 5-10 membered monocyclic or bicyclic aromatic ring is phenyl, pyridyl, pyrazolyl or indolyl and wherein said 5-10 membered monocyclic or bicyclic aromatic ring is optionally substituted with one or more substituents selected from the group consisting of halogens, C _1.4 alkyl, OC _1.4 alkyl, OCF ₃ and OCF2H. 14. The compound of claim 1, wherein the compound is - 179 047929 - 180 047929 - 181 047929 15. A pharmaceutical composition that contains a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14 and that contains at least one pharmaceutically acceptable excipient. 16. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15 as a medicine for the prevention or treatment of an infection caused by HBV (hepatitis B virus) or a disease caused by HBV (hepatitis B virus) in a mammal in need thereof. 17. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15 for the prevention or treatment of an infection caused by HBV (hepatitis B virus) or a disease caused by HBV (hepatitis B virus) in a mammal in need thereof. 18. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15 for the prevention or treatment of chronic hepatitis B. 19. A method for treating an infection caused by HBV (hepatitis B virus) or a disease caused by HBV (hepatitis B virus) in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15.