EA041670B1 - COMPOUND, PHARMACEUTICAL COMPOSITION CONTAINING IT AND METHOD OF TREATMENT OR PREVENTION OF HEPATITIS B VIRUS INFECTION - Google Patents

Description

Related Applications

Application claims priority on U.S. Provisional Application No. 62/550992, filed August 28

2017. The entire disclosure of the above application is incorporated herein by reference.

Technical field

The invention generally relates to new antiviral agents. In particular, the invention relates to compounds that can inhibit the protein(s) encoded by the hepatitis B virus (HBV) or interfere with the functioning of the HBV life cycle, compositions containing such compounds, methods for inhibiting HBV viral replication, methods for treating or preventing HBV infection, and methods for obtaining compounds.

State of the art

HBV infection remains a major public health problem, affecting approximately 2 million people worldwide. Among them, 350 million people worldwide and 1.4 million in the US have a chronic infection that leads to chronic persistent hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). Between 500,000 and 1 million people die each year as a result of end-stage liver disease caused by HBV infection.

Despite the availability of prophylactic HBV vaccines, the burden of chronic HBV infection remains a major unmet global health problem due to suboptimal treatment options and a steady rate of new infections in most parts of the developing world. Current therapies do not provide cure and are limited to only two classes of agents (interferon and nucleoside analogs/viral polymerase inhibitors); drug resistance, poor efficacy, and tolerability issues limit their impact. The low success rate of HBV treatment is at least in part due to the presence and resistance of covalently closed circular DNA (cccDNA) in the nuclei of infected hepatocytes. However, continuous suppression of HBV DNA slows the progression of liver disease and helps prevent HCC. Current therapeutic goals for patients infected with HBV are to reduce serum HBV DNA to low or undetectable levels and ultimately reduce or prevent the development of cirrhosis and HCC.

HBV is an enveloped, partially double-stranded DNA virus (dsDNA) of the Hepadnaviridae family. The HBV capsid or nuclear protein (CP) plays a major role in HBV replication. The dominant biological function of the capsid protein is to act as a structural protein that wraps pregenomic RNA in a protein coat and forms immature capsid particles that spontaneously self-assemble from multiple copies of nuclear dimers in the cytoplasm. The capsid protein also regulates the synthesis of viral DNA through the different phosphorylation status of its C-terminal phosphorylation sites. Also, the capsid protein promotes nuclear translocation of the viral relaxed circular genome via nuclear localization signals centered on the arginine-rich domains of the C-terminal region of the capsid protein. In the nucleus, as a component of the viral cccDNA minichromosome, the capsid protein plays a structural and regulatory role in the functioning of cccDNA minichromosomes. The capsid protein also interacts with the viral protein coat in the endoplasmic reticulum (ER) and initiates the release of the original viral particles from hepatocytes.

Anti-HBV inhibitors having affinity for the capsid have been reported. For example, phenylpropenamide derivatives, including compounds named AT-61 and AT-130 (Feld J. et al., Antiviral Res. 2007, 76, 168), and the class of thiazolidin-4-ones from Valeant (W02006/033995) have been shown to ) inhibit the packaging of pregenomic RNA (pgRNA). Heteroaryldihydropyrimidines or HAPs have been detected in tissue culture screening (Weber et al., Antiviral Res. 2002, 54, 69). These HAP analogs act as synthetic allosteric activators and are able to stimulate the formation of aberrant capsids, which leads to the destruction of the nuclear protein. A subclass of sulfomoylarylamides exhibits activity against HBV (WO2013/006394, WO2013/096744, WO2014/184365 and WO2017/136403). The small molecule bis-ANS has also been shown to act as a molecular wedge and interfere with the development of normal capsid protein geometry and capsid formation (Zlotnick A. et al., J. Virol. 2002, 4848).

There is a need in the art for new therapeutic agents that treat, eliminate or prevent HBV infection. Administration of these therapeutic agents to patients infected with HBV, either alone or in combination with other HBV treatments or adjunctive treatments, results in a significant improvement in prognosis, reduced disease progression, and increased seroconversion rate.

The essence of the invention

This invention relates to new antiviral compounds, pharmaceutical compositions containing such compounds, as well as methods for the treatment or prevention of viral (particularly HBV) infections in a subject in need of such therapy, using these compounds. The compounds of this invention inhibit the protein(s) encoded by the hepatitis B virus (HBV) or interfere with the life cycle of HBV and are also useful as antiviral agents. Additionally, this invention includes a method for obtaining these compounds.

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In its principal aspect, this invention provides a compound of formula (I)

X-A-Y-L-R (I) or a pharmaceutically acceptable salt thereof, wherein X and Y are each independently selected from optionally substituted aryl or optionally substituted heteroaryl; in one embodiment of the invention, one of X and Y is an optionally substituted phenyl; in another embodiment of the invention, both X and Y are optionally substituted phenyl;

A is selected from the group consisting of -NHC(O)-, preferably A is -NHC(O)-;

L is S(O) ₂ , S(O), S or O and

R is connected to L via a carbon atom and is independently selected from the group consisting of optionally substituted -C ₁ -C ₁₀ alkyl, optionally substituted -C ₂ -C ₁₀ alkenyl, optionally substituted C ₂ -C ₁₀ alkynyl, optionally substituted aryl , optionally substituted heteroaryl, optionally substituted -C ₃ -C ₁₂ -cycloalkyl, optionally substituted -C ₃ -C ₁₂ -cycloalkenyl, optionally substituted from 3 to 12-membered heterocyclic fragment; in one embodiment, R is an optionally substituted -C ₅ -C ₁₂ -cycloalkyl or an optionally substituted 5 to 12 membered heterocyclic moiety, each independently substituted with one or more of the following: fused rings, one or more spiro rings, or one or more fragments of a molecule with a bridged ring.

In another embodiment of the invention, R is an optionally substituted C3C ₁₂ -cycloalkyl-C ₁ -C ₆ -alkyl, optionally substituted C ₃ -C ₁₂ -cycloalkenyl-C ₁ -C ₆ -alkyl, or optionally substituted from 3 to 12-membered heterocyclic fragment-C ₁ -C ₆ -alkyl.

Each preferred group listed above may be in combination with one, any or all of the other preferred groups.

Detailed description of the invention

In one embodiment, this invention provides a compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X represents an optionally substituted phenyl. In certain embodiments of the invention, X is phenyl substituted with one or more substituents, namely 1, 2, 3, 4 or 5 substituents. Preferably, the substituents are independently selected from halogen, CN, optionally substituted -C ₁ -C ₃ -alkoxy, optionally substituted -C ₁ -C ₃ -alkyl, and optionally substituted -C ₃ -C ₆ -cycloalkyl. In certain embodiments of the invention, X is phenyl substituted with one or more substituents independently selected from fluorine, chlorine, bromine, methyl, difluoromethyl, trifluoromethyl, CN, and cyclopropyl. In certain embodiments of the invention, X is selected from the groups below

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted monocyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X represents an optionally substituted thiophenyl, an optionally substituted thiazolyl, an optionally substituted pyridyl, or an optionally substituted pyrimidinyl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted pyrimidinyl, optionally substituted pyridazyl or optionally substituted pyrazyl, as follows

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In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted bicyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted 5/6 bicyclic heteroaryl and is connected to A either through a carbon atom or through a nitrogen atom, preferably through a carbon atom, 6-membered ring of said 5/6 bicyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted benzimidazolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinolyl, isoquinolyl or quinazolyl.

In certain embodiments, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted bicyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y represents phenyl, substituted by halogen, CN, optionally substituted -C ₁ -C ₆ -alkoxy, optionally substituted -C1C ₆ -alkyl, optionally substituted -C ₁ -C ₆ -alkenyl, optionally substituted -C ₃ -C ₆ cycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted 1,3 phenylene, for example,

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted 1,3-phenylene, for example,

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted 2,4-pyrrolylene, for example, ¹ . ι or ι.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted monocyclic heteroaryl. In certain embodiments, this invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof, wherein Y is an optionally substituted thiophenyl, an optionally substituted thiazolyl, an optionally substituted pyrrolyl, an optionally substituted pyrazolyl, an optionally substituted imidazolyl, an optionally substituted pyridyl, or an optionally substituted pyrimidinyl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y represents pyrrolyl, optionally substituted with halogen, CN or optionally substituted with -C ₁ -C ₃ -alkyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted bicyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted 5/6 bicyclic heteroaryl and is connected to A either through a carbon atom or through a nitrogen atom of the 5-membered ring of the specified 5 /6 bicyclic heteroaryl. In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where Y is an optionally substituted benzimidazolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinolyl, isoquinolyl or quinazolyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X and Y, each independently, represents an optionally substituted monocyclic heteroaryl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted phenyl, and Y is an optionally substituted monocyclic heteroaryl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X is an optionally substituted monocyclic heteroaryl, and Y is an optionally substituted phenyl.

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In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X and Y, each independently, represents phenyl or monocyclic heteroaryl, each independently substituted with from 1 to 3 substituents selected from the group consisting of halogen, CN, optionally substituted methyl, optionally substituted methoxy, and optionally substituted cyclopropyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X and Y are each independently selected from the group consisting of optionally substituted phenyl, optionally substituted thiophenyl, optionally substituted pyridyl, and optionally substituted pyrimidyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where X and Y, each independently, represents an optionally substituted phenyl. In certain embodiments, this invention relates to compounds of formula (I) and pharmaceutically acceptable salts thereof, wherein X is an optionally substituted phenyl and Y is an optionally substituted pyrrolyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where A represents -NHC(O)-.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts,

CF ₃ /k FF _CF o _FF . ...... „ ________ /L' /I where A is -nhc(0>, n, n or n, and the indicated nitrogen in -nhc(O)-, n, n or n is connected to X.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, and the specified nitrogen in

where A represents connected to Y.

yourself

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where L represents S(O) 2 .

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where L represents S(O).

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where L represents S.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where L represents O.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted C ₁ -C ₁₀ -alkyl, optionally substituted C ₂ -C ₁₀ -alkenyl or optionally substituted C ₂ -C ₁₀ alkynyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted aryl or optionally substituted heteroaryl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted -C ₃ -C ₁₂ -cycloalkyl or optionally substituted from 3 to 12-membered heterocyclic fragment.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted C ₃ -C ₁₂ -cycloalkyl-C ₁ -C ₆ -alkyl, optionally substituted C ₃ -C ₁₂ - cycloalkenyl-C ₁ -C ₆ -alkyl or optionally substituted from 3 to 12-membered heterocyclic -C ₁ -C ₆ -alkyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted -C ₅ -C ₁₂ -cycloalkyl or optionally substituted from 5 to 12-membered heterocyclic fragment, each optionally substituted with one or more fused rings.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted -C ₅ -C ₁₂ -cycloalkyl or optionally substituted from 5 to 12-membered heterocyclic fragment, each optionally substituted with one or more spiro rings.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R represents an optionally substituted -C ₅ -C ₁₂ -cycloalkyl or optionally substituted from 5 to 12-membered heterocyclic fragment, each independently containing a bridging fragment.

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In certain embodiments of the invention, this compound refers to compounds of formula (I), and their pharmaceutically acceptable salts, where

R represents -C(R ₁₀ ) ₃ , and _LI and n in each case independently selected from 0, 1, 2 or 3;

T is in each case independently selected from C(R ₁₀ ) and N;

E is in each case independently selected from -C(R ₁₀ ) ₂ -, -N(R ₁₀ )-, O, S, S(O) and S(O) ₂ ;

and R ₁₀ in each case independently selected from the group consisting of hydrogen, halogen, -CN, -NO ₂ , optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₂ -C ₈ -alkenyl, optionally substituted -C ₂ -C ₈ -alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8-membered heterocyclic fragment, optionally substituted aryl, optionally substituted heteroaryl, and -L1-R1;

wherein L1 is -O-, -S-, -NR _r , -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R1)-, - N(Ri)C(O)-, -OC(O)N(Ri)-, -N(R1)C(O)O-, -N(R ₁ )C(O)N(R ₁ )-, -S(O)-, -S(O)2-, -S(OhN(R1)-, -N(Ri)S(O)2-;

R1 is at each occurrence independently selected from the group consisting of hydrogen, optionally substituted -C ₁ -C ₆ alkyl, optionally substituted -C ₂ -C ₈ alkenyl, optionally substituted -C ₂ -C ₈ alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8 membered heterocyclic fragment, optionally substituted aryl and optionally substituted heteroaryl.

In certain embodiments, each R ₁₀ is independently selected from hydrogen, halogen, hydroxy, protected hydroxy, -CN, -NO ₂ , amino, protected amino, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₁ -C ₆ -alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and -O-(prodrug hydroxyl group). In certain embodiments of the invention, the prodrug hydroxyl group is a phosphate or sulfamate. In certain embodiments of the invention, said prodrug hydroxyl group is an acyl group derived from an amino acid, preferably an α-amino acid.

In certain embodiments of the invention, each R ₁₀ is independently optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, protected hydroxy, amino, protected amino, and optionally substituted heteroaryl .

In certain embodiments of the invention, two adjacent R ₁₀ groups, together with the carbon or nitrogen atoms to which they are attached, form an olefinic or imine double bond or a fused ring. In certain embodiments, the two R ₁₀ geminal groups together form an oxo, optionally substituted olefin, optionally substituted oxime, or spiro ring. In certain embodiments of the invention, the two remote R ₁₀ groups, together with the atoms to which they are attached, and any intermediate atoms, form a bridge fragment.

In certain embodiments of the invention, this invention relates to compounds of formula (I) and their pharmaceutically acceptable salts, where R is optionally substituted -(CH ₂ ) _0-4 -C(R ₁₀ ) ₃ , _or where n, E, T and R ₁₀ previously described;

v choose from 1, 2, 3 or 4.

In certain embodiments, each R ₁₀ is independently selected from hydrogen, halogen, hydroxy, protected hydroxy, -CN, -NO ₂ , amino, protected amino, optionally substituted -Cl-C ₆ -alkyl, optionally substituted -C ₁ -C ₆ -alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and -O-(prodrug hydroxyl group). In certain embodiments of the invention, the prodrug hydroxyl group is a phosphate or sulfamate. In certain embodiments of the invention, said prodrug hydroxyl group is an acyl group derived from an amino acid, preferably an α-amino acid. In certain embodiments, each R ₁₀ is independently -Cl-C ₆ -alkyl optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, protected hydroxy, amino, protected amino, and optionally substituted heteroaryl. In certain embodiments, two adjacent R ₁₀ groups, together with the carbon or nitrogen atoms to which they are attached, form an olefinic double bond, an imine double bond, or a fused carbocyclic or heterocyclic ring. In certain embodiments, the two R ₁₀ geminal groups together form an oxo, optionally substituted olefin, optionally substituted oxime, or spiro ring. In certain embodiments of the invention, the two removed R ₁₀ groups, together with the atoms to which they are connected, and any intermediate atoms form a bridge fragment.

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In certain embodiments of the invention, R is selected from the groups below and is optionally substituted

In certain embodiments of the invention, R is selected from the groups below and is optionally substituted

In another embodiment, the compound of formula (I) is represented by formulas (Ia)(Ic) or (Id) or a pharmaceutically acceptable salt thereof:

where X, A, Y and R are previously described.

In another embodiment, the compound of formula (I) is represented by formulas (IIa)(IIg) or (IIh) or a pharmaceutically acceptable salt thereof

where X, A, Y and R are previously described.

In certain embodiments of the invention, this invention relates to compounds of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X and Y, each independently, represents an optionally substituted phenyl or optionally substituted monocyclic heteroaryl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X and Y, each independently, represents an optionally substituted phenyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X is an optionally substituted phenyl, and Y is an optionally substituted 5 -membered heteroaryl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X is an optionally substituted 5-membered heteroaryl, and Y is optionally substituted phenyl.

In certain embodiments of the invention, this invention relates to compounds

- 6 041670 of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X is an optionally substituted phenyl and Y is an optionally substituted pyrrolyl.

In certain embodiments of the invention, this invention relates to compounds of formula (I) represented by formulas (IIa)-(IIg) or (IIh), or their pharmaceutically acceptable salts, where X and Y, each independently, represents an optionally substituted phenyl, optionally substituted naphthyl, optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted thiophenyl, optionally substituted pyrrolyl, optionally substituted thiazolyl, optionally substituted thiadiazolyl, optionally substituted oxazolyl, optionally substituted isoxazolyl, optionally substituted oxadiazolyl, optionally substituted imidazolyl, optionally substituted pyrazolyl, optionally substituted triazolyl or optionally substituted quinolinyl.

In another embodiment, the compound of formula (I) is represented by formulas (IIIa)-(IIIc) or (IIId) or a pharmaceutically acceptable salt thereof

where m in each case is independently 0, 1, 2, 3 or 4;

R ₁₄ is independently selected at each occurrence from the group consisting of hydroxy, protected hydroxy, halogen, -CN, -NO ₂ , optionally substituted amino, N ₃ , optionally substituted aryl, optionally substituted heteroaryl, optionally substituted -C ₁ -C ₆ - alkyl, optionally substituted -C ₂ -C ₈ -alkenyl, optionally substituted -C ₂ -C ₈ -alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8-membered heterocyclic fragment, optionally substituted -C ₁ -C ₆ -alkoxy, -C (O) ₂ -C ₁ -C ₆ -alkyl, -C (O) NH-C ₁ -C ₆ -alkyl and -C (O) -C ₁ -C ₆ -alkyl ; And

R has been previously described.

In another embodiment, the compound of formula (I) is represented by (IIIa-1)-(IIIc-1) or (IIId-1) or a pharmaceutically acceptable salt thereof

where X, R, R ₁₄ and m previously described.

In another embodiment, the compound of formula (I) is represented by (IIIa-2)-(IIIc-2) or (IIId-2) or a pharmaceutically acceptable salt thereof:

where Y, R, R ₁₄ and m previously described.

In another embodiment of the invention, the compound of formula (I) is represented by (IVa)-(IVd) or (IVe) or a pharmaceutically acceptable salt thereof, formulas formulas formulas

where E, T, m, n, R ₁₀ and R ₁₄ previously described.

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In another embodiment, the compound of formula (I) is represented by formulas (Va)(Vd) or (Ve) or a pharmaceutically acceptable salt thereof

where ml in each case independently represents 1, 2 or 3;

m2 is independently 0, 1, or 2 at each occurrence;

m3 is independently 0, 1, 2, or 3 at each occurrence;

R ₂₁ is at each occurrence independently selected from the group consisting of halo, -CN, optionally substituted -C ₁ -C ₆ alkyl, optionally substituted -C ₁ -C ₆ alkoxy, and optionally substituted -C ₃ -C ₈ cycloalkyl;

R ₂₂ is independently selected at each occurrence from the group consisting of halo, -CN, optionally substituted -C ₁ -C ₆ alkyl, optionally substituted -C ₁ -C ₆ alkoxy, optionally substituted aryl, and optionally substituted heteraryl; And

E, n and R ₁₀ previously described.

In another embodiment, a compound of formula (I) is represented by formulas (Vf)(Vh) or (Vj) or a pharmaceutically acceptable salt thereof

where m4 in each case is independently 0, 1 or 2;

R ₃₀ is hydrogen, optionally substituted with -C ₁ -C ₆ alkyl, a hydroxy protecting group, or an inactive hydroxy group;

m1, m2, n, E, R ₁₀ , R ₂₁ and R ₂₂ have been previously described.

In certain embodiments of the invention, R ₃₀ is a phosphate or a sulfamate. In certain embodiments of the invention, R ₃₀ is an acyl group derived from an amino acid, preferably an α-amino acid.

In another embodiment, the compound of formula (I) is represented by formulas (VIa)-(VIe) or (VIf) or a pharmaceutically acceptable salt thereof

where m1 in each case independently represents 1, 2 or 3;

m2 is independently 1 or 2 at each occurrence;

n, R ₂₁ , R ₂₂ and R ₃₀ have been previously described.

In certain embodiments of the invention, R ₃₀ is a phosphate or a sulfamate. In certain embodiments of the invention, R ₃₀ is an acyl group derived from an amino acid, preferably an α-amino acid.

In another embodiment of the invention, the compound of formula (I) is represented by the formulas

- 8 041670 (VIa)-(VIe) or (VIf) or its pharmaceutically acceptable salt, where m1 in each case independently represents 2 or 3;

m2 in each case is 1;

n is at each occurrence independently 0, 1 or 2;

R ₂₁ is halo, CN, optionally substituted methyl, optionally substituted methoxy, and optionally substituted cyclopropyl;

R22 is halo, CN, optionally substituted methyl and optionally substituted methoxy;

R ₃₀ is an acyl group derived from an amino acid.

In certain embodiments of the invention, R ₃₀ is an acyl group derived from an α-amino acid containing an aliphatic side chain. In certain embodiments of the invention, R30 is an acyl group derived from alanine or valine.

In another embodiment, the compound of formula (I) is represented by formulas (VIIa)-(VIId) or (VIIe) or a pharmaceutically acceptable salt thereof

(Vile) where m1, m3, n, E, R ₁₀ , R ₂₁ and R ₂₂ previously described.

In another embodiment, the compound of formula (I) is represented by formulas (VIIa-1)-(VIId-1) or (VIIe-1) or a pharmaceutically acceptable salt thereof

where R ₂₁ at each occurrence is independently selected from the group consisting of hydrogen, halogen, -CN, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₁ -C ₆ -alkoxy and optionally substituted C ₃ -C ₈ - cycloalkyl;

m3, n, E, R ₁₀ , R ₂₁ and R ₂₂ have been previously described.

In another embodiment, the compound of formula (I) is represented by formulas (VIIa-2)-(VIIc-2) or (VIId-2) or a pharmaceutically acceptable salt thereof

where m4, n, E, R ₁₀ , R ₂₁ , R ₂₂ , R ₂₃ and R ₃₀ have previously been described.

In certain embodiments of the invention, R ₃₀ is hydrogen. In certain embodiments of the invention, R ₃₀ is an acyl group derived from an amino acid, preferably an α-amino acid.

In another embodiment, the compound of formula (I) is represented by formulas (VIIa-2)-(VIIc-2) or (VIId-2) or a pharmaceutically acceptable salt thereof, where n is independently 0, 1 or 2 at each occurrence;

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R ₂₁ is independently at each occurrence halogen, CN, optionally substituted methyl, optionally substituted methoxy, or optionally substituted cyclopropyl;

R ₂₂ is halo, CN, optionally substituted methyl, or optionally substituted methoxy;

R ₂₃ is hydrogen or halogen;

R ₁₀ represents hydrogen, halogen, hydroxyl group or optionally substituted C ₁ -C ₆ -alkyl;

R ₃₀ is hydrogen or an acyl group derived from an amino acid.

In certain embodiments of the invention R ₂₁ in each case represents fluorine. In certain embodiments of the invention R ₂₂ represents fluorine or chlorine. In certain embodiments, R ₁₀ is hydrogen, halogen, hydroxyl, C ₁ -C ₆ alkyl optionally substituted with one or more groups selected from halogen, hydroxy, or optionally substituted C ₁ -C ₆ alkoxy. In certain embodiments of the invention R ₂₃ represents fluorine. In certain embodiments of the invention, R ₃₀ is an acyl group derived from alanine or valine.

In another embodiment, the compound of formula (I) is represented by formulas (VIIIa)-(VIIIc) or (VIIId) or a pharmaceutically acceptable salt thereof

where R11 is at each occurrence independently selected from the group consisting of hydrogen, halogen, hydroxy, protected hydroxy, -CN, amino, protected amino, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₂ -C8 -alkenyl, optionally substituted -C ₂ -C8 -alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8-membered heterocyclic fragment, optionally substituted -C ₁ -C ₆ -alkoxy , optionally substituted -NH-C ₁ -C ₆ -alkyl, optionally substituted -X(C ₁ -C ₆ -alkyl) ₂ , -CO ₂ H, optionally substituted -C (O) ₂ -C _{1 -C 6} -alkyl , optionally substituted -C(O)NH-C ₁ -C ₆ -alkyl and optionally substituted -C(O)-C ₁ -C ₆ -alkyl; m, n, E, T, R ₁₀ and R ₁₄ previously described. In certain embodiments, preferred R11 groups include hydrogen, halogen, hydroxy, protected hydroxy, protected amino, optionally substituted aryl, optionally substituted heteroaryl, -CO ₂ H, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted NH(HC (O) ₂ -C ₁ -C ₆ -alkyl and optionally substituted -C ₁ -C ₆ -alkoxy In certain embodiments, each Rn is optionally substituted -C ₁ -C ₆ -αkyl, optionally substituted aryl, optionally substituted heteroaryl; and optionally substituted -C ₁ -C ₆ -alkoxy.

In another embodiment, the compound of formula (I) is represented by formulas (VIIIa-1)-(VIIIc-1) or (VIIId-1) or a pharmaceutically acceptable salt thereof

where m, n, E, R ₁₀ , R11 and R ₁₄ previously described.

In another embodiment, the compound of formula (I) is represented by formulas (IXa)-(IXc) or (IXd) or a pharmaceutically acceptable salt thereof

where m4, n, R ₁₀ , R ₂₁ , R ₂₂ , R ₂₃ and R ₃₀ have been previously described.

In another embodiment, the compound of formula (I) is represented by formulas (Xa)- 10 041670 (Xc) or (Xd) or a pharmaceutically acceptable salt thereof

where m4, n, R ₁₀ , R ₂₁ , R ₂₂ , R ₂₃ and R ₃₀ have been previously described.

In another embodiment, a compound of formula (I) is represented by formulas (XIa)-(XIc) or (XId) or a pharmaceutically acceptable salt thereof

where m4, n, R ₁₀ , R ₂₁ , R ₂₂ , R ₂₃ and R ₃₀ have been previously described.

In another embodiment, the compound of formula (I) is represented by formulas (XIIa)-(XIIc) or (XIId) or a pharmaceutically acceptable salt thereof

where R ₃₁ is at each occurrence independently selected from the group consisting of hydrogen, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₂ -C ₈ -alkenyl, optionally substituted -C ₂ -C ₈ -alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8-membered heterocyclic fragment;

m1, m2, m4, n, R ₁₀ , R11, R ₂₁ , R ₂₂ and R ₃₀ have been previously described.

In certain embodiments, R ₃₁ is C ₁ -C ₆ alkyl optionally substituted with one or more groups selected from halogen, hydroxy, optionally substituted C ₁ -C ₆ alkoxy, amino, optionally substituted -NH-C ₁ - C ₆ -alkyl, optionally substituted -N(C ₁ -C ₆ -alkyl) ₂ , optionally substituted -CO ₂ -C ₁ -C ₆ -alkyl, optionally substituted -C (O)NH-C ₁ -C ₆ -alkyl , optionally substituted -NHC(O)-C ₁ -C ₆ -alkyl, optionally substituted -C(O)-C1-C ₆ -alkyl and optionally substituted -NHS(O) ₂ -C ₁ -C ₆ -alkyl.

In another embodiment, the compound of formula (I) is represented by formulas (XIIIa)-(XIIIc) or (XIIId) or a pharmaceutically acceptable salt thereof

where R ₃ 2 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted C ₁ -C6 alkyl, optionally substituted -C(O)-C ₁ -C ₆ alkyl, optionally substituted -C(O)-C2 -C ₈ -alkenyl, optionally substituted -C(O)-C ₂ -C ₈ -alkynyl, optionally substituted C(O)-C ₃ -C ₈ -cycloalkyl, optionally substituted -C(O)aryl, optionally substituted -C (O)-(38-membered heterocyclic fragment), optionally substituted -C(O)-C ₁ -C ₆ -alkyl, optionally substituted -CO ₂ -C ₁ -C ₆ -alkyl, optionally substituted S(O) ₂ - C ₁ -C ₆ -alkyl, optionally substituted with S(O) ₂ -C ₂ -C ₆ -alkenyl; preferably one R ₃₂ is optionally substituted -C(O)-C ₁ -C ₆ -alkyl, optionally substituted -C(O)-C ₂ -C ₈ -alkenyl, optionally substituted -C(O )-C ₂ -C ₈ -alkynyl, optionally substituted -C (O) -C ₃ -C ₈ -cycloalkyl, optionally substituted -C (O) aryl, optionally substituted -C (O) - (3-8-membered heterocyclic fragment), optionally substituted -C(O)-C ₁ -C ₆ -alkyl, optionally substituted -CO ₂ -C ₁ -C ₆ -alkyl, optionally substituted S(O) ₂ -C ₁ -C ₆ -alkyl, optionally substituted 8(O) ₂ -C ₂ -C ₆ -alkenyl, then the other R ₃₂ is hydrogen or optionally substituted C ₁ -C ₆ -alkyl;

m1, m2, m4, n, R ₁₀ , R11, R ₂₁ , R ₂₂ and R ₃₀ have been previously described.

In certain embodiments of the invention, the two R ₃₂ groups, together with the nitrogen atom to which they are attached, form a 3 to 8-membered heterocyclic ring.

In another embodiment, a compound of formula (I) is represented by formulas (XIVa)-(XIVc) or (XIVd) or a pharmaceutically acceptable salt thereof

where R ₃₃ is at each occurrence independently selected from the group consisting of hydrogen, optionally substituted -C ₁ -C ₆ alkyl, optionally substituted -C ₂ -C ₈ alkenyl, optionally substituted C ₂ -C ₈ alkynyl, optionally substituted - C ₃ -C ₈ -cycloalkyl, optionally substituted from 3 to 8-membered heterocyclic fragment, optionally substituted -C ₁ -C ₆ -alkoxy, optionally substituted -NH-C ₁ -C ₆ -alkyl, optionally substituted -N(C ₁ -C ₆ -alkyl) ₂ , optionally substituted -NH-C ₁ -C ₆ -alkenyl, optionally substituted -NH-(3-8-membered heterocyclic fragment);

m1, m2, m4, n, R ₁₀ , R _n , R ₂₁ , R ₂₂ and R ₃₀ have been previously described.

In another embodiment, the compound of formula (I) is represented by formulas (XVa)-(XVc) or (XVd) or a pharmaceutically acceptable salt thereof

where R34 is at each occurrence independently selected from the group consisting of hydrogen, optionally substituted -C ₁ -C ₆ -alkyl, optionally substituted -C ₂ -C ₈ -alkenylα, optionally substituted C ₂ -C ₈ alkynyl, optionally substituted -C ₃ -C ₈ -cycloalkylα and optionally substituted from 3 to 8-membered heterocyclic fragment;

m1, m2, m4, n, R ₁₀ , Rn, R ₂₁ , R ₂₂ , R ₃₀ and R ₃₁ have been previously described.

In another embodiment, the compound of formula (I) is represented by formula (XVI) or a pharmaceutically acceptable salt thereof

where R ₂₁ , R21' and R ₂₁ are independently selected from hydrogen, fluorine, methyl, difluoromethyl and trifluoromethyl and

R ₃ 5 is -[CH(R ₃₆ )]pC(R ₃₇ )(R ₃₈ )OH or -CH2-O-CH2-[CH(R ₃₆ )]pC(R ₃₇ )(R ₃₈ )OH, where p is 0 or 1;

R ₃₆ represents hydrogen, methyl or hydroxyl group; a

R ₃₇ and R ₃₈ are independently hydrogen or methyl.

Preferably at least two of R ₂₁ , R ₂₁ ' and R ₂₁ are not hydrogen. More preferably (1) none of R ₂₁ , R ₂₁ ' and R ₂₁ is hydrogen; or (2) R ₂₁ is hydrogen and R ₂₁ ' and R ₂₁ are not hydrogen. Preferably at least two of R ₂₁ , R ₂₁ ' and R ₂₁ are fluorine. In other embodiments of the invention, each of R ₂₁ , R ₂₁ ' and R ₂₁ represents fluorine.

- 12 041670

In another embodiment, the compound of formula (I) is represented by formula (XVII) or a pharmaceutically acceptable salt thereof

where R21, R21' and R21 are independently selected from hydrogen, fluorine, methyl, difluoromethyl and trifluoromethyl; And

R39 is a hydroxyl group and

R ₄₀ is -[C(R ₄₁ )(R ₄₂ )] _q -R ₄₃ where q is 2;

R ₄₁ is hydrogen and R42 is hydroxyl;

or, alternatively, R41 and R ₄₂ of the same C(R ₄₁ )(R ₄₂ ) group together may form oxo; And

R ₄₃ is C ₁ -C ₆ alkyl.

It is clear that the description of the invention in this text is to be construed in accordance with the laws and principles of chemical bonding. In some examples, a hydrogen atom must be substituted to attach a substituent to any given site.

It is also understood that the compounds of this invention may contain one or more asymmetric carbon atoms and may exist in racemic, diastereometric or optically active form. It is also understood that certain compounds of this invention may exist in various tautomeric forms. All tautomers are intended to be within the scope of this invention.

In one aspect, the compounds of the invention are useful in the treatment of HBV by disrupting, accelerating, reducing, slowing down and/or inhibiting the normal function of the viral core protein, including, but not limited to, direct or indirect interaction with the relaxed circular (pk) DNA of the virus, cccDNA and reverse transcriptase, direct or indirect interaction with host proteins, namely with histones, or with host partners, namely kinase, assembly and / or disassembly of mature or immature viral capsid protein particles, thus stimulating the construction of an aberrant capsid and leading to an antiviral effect, namely, disruption of assembly and/or disassembly of virions, maturation of virions and/or release of the virus. In one embodiment, the capsid assembly disruptor interacts with the mature or immature viral capsid and disrupts the stability of the capsid, thereby affecting assembly and/or disassembly. In another embodiment, the capsid assembly disruptor disrupts protein folding and/or salt bridges necessary for the stability, function and/or normal morphology of the viral capsid, hence disrupting and/or accelerating capsid assembly and/or disassembly. In yet another embodiment of the invention, the compounds of the invention bind the capsid and alter the metabolism of cellular polyproteins and precursors, leading to excessive accumulation of protein monomers and/or oligomers and/or abnormal particles, which causes cellular intoxication and death of infected cells. In another embodiment, the compounds of the invention result in a failure of capsid formation of optimal stability, affecting efficient shedding and/or disassembly of viruses (eg, during infectivity).

In one embodiment, the compounds of the invention disrupt and/or accelerate assembly and/or disassembly of the capsid when the capsid protein is immature. In another embodiment, the compounds of the invention disrupt and/or accelerate assembly and/or disassembly of the capsid when the capsid protein is mature. In yet another embodiment of the invention, the compounds of the invention disrupt and/or accelerate the assembly and/or disassembly of the capsid during viral infectivity. In yet another embodiment of the invention, disruption and/or acceleration of capsid assembly and/or disassembly attenuates HBV viral infectivity and/or reduces viral load. In yet another embodiment of the invention, disruption, acceleration, inhibition, retardation, and/or reduction in assembly and/or disassembly of the capsid destroys the virus in the host. In yet another embodiment of the invention, the compounds of the invention disrupt and/or reduce the frequency of interaction between the nuclear protein and viral rcDNA, cccDNA or reverse transcriptase during viral infectivity. In yet another embodiment of the invention, the compounds of the invention disrupt and/or reduce the frequency of interaction between a nuclear protein and partners or host proteins during viral infectivity. In another embodiment of the invention, the destruction of HBV in the host body successfully eliminates the need for chronic long-term therapy and/or reduces the duration of long-term therapy.

In one embodiment, the compounds described herein are suitable for monotherapy and are effective against naturally occurring and related HBV strains and against currently known drug resistant HBV strains. In another implementation

- 13 041670 of the invention the compounds described in this document are suitable for use in combination therapy.

In another embodiment, the compounds of the invention are used in methods of altering (eg, to inhibit, disrupt, or accelerate) HBV cccDNA activity. In yet another embodiment of the invention, the compounds of the invention are used in methods for reducing or preventing the formation of HBV cccDNA. In another embodiment of the invention, the additional therapeutic agent is selected from immunomodulator or immunostimulant preparations that include the T-cell immune response activator AIC649, and biological agents belonging to the class of interferons, namely interferon alfa 2a or 2b, or modified interferons, namely pegylated interferon, alpha 2a, alpha 2b, lambda; or STIN modulators (stimulators of interferon genes); or TLR modulators, namely TLR-7 agonists or TLR-9 agonists; or therapeutic vaccines for stimulating a specific immune response to HBV, namely virus-like particles consisting of HBcAg and HBsAg, immune complexes of HBsAg and HBsAb, or recombinant proteins containing HBx, HBsAg and HBcAg in the context of a yeast vector; or immunoactivators, namely SB-9200 certain cellular viral RNA sensors, namely RIG-I, NOD2 and MDA5 proteins; or RNA interference (mRNA) or small interfering RNA (siRNA), namely ARC-520, ARC-521, ARB-1467 and ALN-HBV mRNA; or other nuclear protein inhibitors or modulators; or antiviral agents that block viral exit or maturation or interfere with HBV polymerase, such as nucleoside or nucleotide or non-nucleo(t)ide polymerase inhibitors, and agents by a specific or unknown mechanism, including agents that interfere with the functioning of other important (s) viral(s) protein(s) or host proteins required for HBV replication or resistance, namely REP 2139 and RG7834. In an embodiment of combination therapy, the reverse transcriptase inhibitor is at least one of zidovudine, didanosine, zalcitabine, DAA, stavudine, lamivudine, abacavir, emtricitabine, entecavir, apricitabine, atevirapine, ribavirin, aciclovir, famciclovir, valaciclovir, ganciclovir, valganciclovir, tenofovir, adefovir, PMRA, cidofovir, efavirenz, nevirapine, delavirdine, or etravirine.

In another 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), AZD 8848 (methyl[3-({[3-( 6amino-2-butoxy-8-oxo-7,8-dihydro-9H-purin-9-yl)propyl][3-(4-morpholine)propyl]amino-1-methyl)phenyl]acetate), GS-9620 (4-amino-2-butoxy-8-[3-(1-pyrrolidinylmethyl)benzyl]-7,8-dihydro-6-(5Hpteridinone) and RO6864018.

In another embodiment of the combination therapy, the TLR-7 agonist is GS9688.

In an embodiment of these combination therapies, a compound and an additional therapeutic agent are combined. In another embodiment, the compound and the additional therapeutic agent are administered together.

In another embodiment of combination therapy, administering a compound of the invention allows the administration of an additional therapeutic agent at a lower dosage or frequency, compared to administering at least one additional therapeutic agent alone, which is necessary to achieve similar results in the prophylactic treatment of HBV infection in an individual in need.

In another embodiment of the combination therapy, before administering a therapeutically effective amount of a compound of the invention, it is verified that the individual is resistant to a compound selected from the group consisting of an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a virus maturation inhibitor, a certain capsid assembly modulator, antiviral compounds of a specific or unknown mechanism and combinations thereof.

In yet another embodiment of the method of the invention, administering a compound of the invention reduces the viral load in an individual to a greater extent than administering a compound selected from the group consisting of an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a virus maturation inhibitor, a certain capsid modulator. assembly, antiviral compounds of a specific or unknown mechanism, and combinations thereof.

In another embodiment, administering a compound of the invention reduces the incidence of virus mutation and/or viral resistance, as compared to administering a compound selected from the group consisting of an HBV polymerase inhibitor, an interferon, a viral entry inhibitor, a virus maturation inhibitor, a certain capsid assembly modulator. , antiviral compounds of a specific or unknown mechanism and combinations thereof.

It should be understood that the compounds covered by this invention are suitable for use as pharmaceuticals.

Definitions.

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

The term aryl as used herein refers to a mono- or polycyclic carbocyclic ring system containing at least one aromatic ring, including but not limited to phenyl, naphthyl, tetrahydronaphthyl, idanyl, and indenyl. A polycyclic aryl is a polycyclic ring system that contains at least one aromatic ring. Polycyclic aryls contain fused rings, covalently linked rings, or combinations thereof.

The term heteroaryl as used herein refers to a mono- or polycyclic aromatic radical having one or more ring-forming atoms selected from S, O and N; and the remaining ring forming atoms are carbon, any N or S contained in the ring being optionally oxidized. Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. Polycyclic heteroaryl contains fused rings, covalently linked rings, or a combination thereof.

In accordance with the invention, aromatic groups may be substituted or unsubstituted.

The term bicyclic aryl or bicyclic heteroaryl refers to a ring system consisting of two rings, at least one ring being aromatic; and two rings are fused or covalently bonded.

The term alkyl as used herein refers to saturated, straight or branched hydrocarbon radicals. C ₁ -C ₃ -alkyl, C ₁ -C6 -alkyl, C ₁ -C ₁ o-alkyl, C ₂ -C ₄ -alkyl or C ₃ -C ₆ -alkyl refer to alkyl groups containing from one to three, one to six, one to ten carbon atoms, 2 to 4 and 3 to 6 carbon atoms, respectively. Examples of C1-C8 alkyl radicals include, but are not limited to, radicals such as methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, neopentyl, n-hexyl, heptyl, and octyl.

The term alkenyl as used herein refers to linear or branched hydrocarbon radicals having at least one carbon-carbon double bond formed by the removal of one hydrogen atom. C ₂ -C ₁₀ -Alkenyl, C ₂ -C ₈ -alkenyl, C ₂ -C ₄ -alkenyl or C ₃ -C ₆ -alkenyl refer to alkenyl groups containing two to ten, two to eight, two to four or three to six carbon atoms, respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

The term alkynyl as used herein refers to linear or branched hydrocarbon radicals having at least one carbon-carbon triple bond formed by the removal of one hydrogen atom. C2-C1o alkynyl, C ₂ -C ₈ alkynyl, C ₂ -C ₄ alkynyl, or C ₃ -C ₆ alkynyl refers to alkynyl groups containing two to ten, two to eight, two to four, or three to six carbon atoms, respectively. Exemplary alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

The term cycloalkyl as used herein refers to a monocyclic or polycyclic saturated carbocyclic ring, or a bi- or tricyclic group in a fused, bridged, or spiro system, and the carbon atoms may be optionally oxo-substituted or optionally substituted with an exocyclic olefinic, imine, or oxime double bond . Preferred cycloalkyl groups include C ₃ -C ₁₂ cycloalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₈ cycloalkyl and C ₄ -C ₇ cycloalkyl. Examples of C ₃ -C ₁₂ -cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 4methylenecyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl , 3methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, etc.

The term cycloalkenyl as used herein refers to a monocyclic or polycyclic carbocyclic ring or a bi- or tricyclic group in a fused, bridged or spiro system having at least one carbon-carbon double bond and the carbon atoms may optionally be oxo- substituted or optionally substituted with an exocyclic olefinic, imine or oxime double bond. Preferred cycloalkenyl groups include C ₃ -C ₁₂ cycloalkenyl, C ₃ -C ₈ cycloalkenyl or C ₅ -C ₇ cycloalkenyl. Examples of C ₃ -C ₁₂ -cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl , spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl, bicyclo[4.2.1]non-3-en-9-yl, etc.

As used herein, the term arylalkyl means a functional group in which an alkylene chain is attached to an aryl group, for example -CH ₂ CH ₂ -phenyl. The term substituted arylalkyl means an arylalkyl functional group in which the aryl group is substituted. Similarly, the term heteroarylalkyl refers to a functional group in which an alkylene chain is bonded to a heteroaryl group. The term "substituted heteroarylalkyl" means a heteroarylalkyl functional group in which the heteroaryl group is substituted.

As used herein, the term alkoski, alone or in combination with other terms, means, unless otherwise stated, an alkyl group having the specified number of carbon atoms connected to the rest of the molecule via an oxygen atom, namely, for example, methoxy, ethoxy, 1-propoxy-, 2-propoxy (isopropoxy) and higher homologues and isomers. Preferred alkoxy are (C ₁ -C ₃ )alkoxy.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, and cycloalkenyl moiety described herein is also an aliphatic group or an alicyclic group.

An aliphatic group is a non-aromatic fragment of a molecule, consisting of any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contains one or more sites of unsaturation, such as double and/or triple bonds. Examples of aliphatic groups are functional groups, namely alkyl, alkenyl, alkynyl, O, OH, NH, NH2, C(O), S(O)2, C(O)O, C(O)NH, OC(O) O, OC(O)NH, OC(O)NH ₂ , S(O) ₂ NH, S(O) ₂ NH ₂ , NHC(O)NH ₂ , NHC(O)C(O)NH, NHS(O ) ₂ NH, NHS(O) ₂ NH ₂ , C(O)NHS(O) ₂ , C(O)NHS(O) ₂ NH or C(O)NHS(O) ₂ NH ₂ etc., groups containing one or more functional groups, non-aromatic hydrocarbons (optionally substituted), and groups in which one or more carbon in the non-aromatic hydrocarbon (optionally substituted) is replaced by a functional group. The carbon atoms in the aliphatic group are optionally oxo-substituted. The aliphatic group is linear, branched, cyclic, or a combination thereof, and preferably contains from about 1 to about 24 carbon atoms, more specifically, from about 1 to about 12 carbon atoms. In addition to the aliphatic hydrocarbon groups used herein, aliphatic groups intentionally include, for example, alkoxyalkyls, polyalkoxyalkyls, such as polyalkylene glycols, polyamines, and polyimines, for example. Aliphatic groups are optionally substituted.

The terms heterocyclic or heterocycloalkyl are used interchangeably and refer to a non-aromatic ring or a bi- or tricyclic group in a fused, bridged, or spiro system, wherein (1) each ring system contains at least one heteroatom independently selected from oxygen, sulfur, and nitrogen , (2) each ring system is saturated or unsaturated, (3) the nitrogen and sulfur heteroatoms are optionally oxidized, (4) the nitrogen heteroatom is optionally quaternary, (5) any of the above rings is fused to an aromatic ring, and (6) the remaining ring-forming atoms are are carbon atoms which are optionally oxo-substituted or optionally substituted with an exocyclic olefinic, imine or oxime double bond. Exemplary heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isooxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinolyl, 2-azabicyclo[ 1]heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 1-ox-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl, and tetrahydrofuryl. Such heterocyclic groups are further substituted. Heteroaryl or heterocyclic groups are attached to the C or N atom (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphatic moiety and the like described herein is also a divalent or multivalent group when used as a link for compounds of two or more groups or substituents that are on the same atom or on different atoms. One skilled in the art will readily determine the valency of any such group from the context in which it is used.

The term substituted refers to substitution by independent replacement of one, two or three or more hydrogen atoms with substituents including, but not limited to, -F, -Cl, -Br, -I, -OH, C ₁ -C ₁₂ -alkyl; C ₂ -C ₁₂ -alkenyl, C ₂ -C ₁₂ -alkynyl, protected hydroxy group, -NO ₂ , -N ₃ , -CN, -NH2, protected amino group, oxo, thioxo, -NH-C ₁ -C ₁₂ -alkyl , -NH-C ₂ -C ₈ -alkenyl, -NH-C ₂ -C ₈ -alkynyl, -NHC ₃ -C ₁₂ -cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocycloalkyl, dialkylamino, diarylamino , diheteroarylamino, -O-C1-C ₁₂ -alkyl, -OC ₂ -C ₈ -alkenyl, -O-C ₂ -C ₈ -alkynyl, -O-C ₃ -C ₁₂ -cycloalkyl, -O-aryl, - O-heteroaryl, -O-heterocycloalkyl, -C (O) -C1-C ₁₂ -alkyl, -C (O) -C ₂ -C ₈ -alkenyl, -C (O) -C ₂ -C ₈ -alkynyl, -C (O) -C ₃ -C ₁₂ -cycloalkyl, -C (O) aryl, -C (O) -heteroaryl, -C (O) -heterocycloalkyl, -CONH2, -CONH-C ₁ -C ₁₂ -alkyl , -CONH-C ₂ -C ₈ -alkenyl, -CONH-C ₂ -C ₈ -alkynyl, -CONH-C ₃ -C ₁₂ -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, - OCO ₂ -C ₁ -C ₁₂ -alkyl, -OCO ₂ -C ₂ -C ₈ -alkenyl, -OCO ₂ -C ₂ -C ₈ -alkynyl, -OCO ₂ -C ₃ -C ₁₂ -cycloalkyl, -OCO ₂ -aryl, -OCO ₂ -heteroaryl, -OSO ₂ -heterocycloalkyl, -CO ₂ -C ₁ -C ₁₂ -alkyl, -CO ₂ -C ₂ -C ₈ -alkenyl, -CO ₂ - C ₂ -C ₈ -alkynyl, CO ₂ -C ₃ -C ₁₂ -cycloalkyl, -CO2 aryl, CO ₂ -heteroaryl, CO ₂ -heterocycloalkyl, -OCONH2, -OCONH-C ₁ -C ₁₂ -alkyl, -OCONH-C2 -C8alkenyl, -OCONH-C ₂ -C ₈ -alkynyl, -OCONH-C ₃ -C ₁₂ -cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl,

- 16 041670

-OCONH-heterocycloalkyl, -NHC(O)H, -NHC(O)-C ₁ -C ₁₂ -alkyl, -NHC(O) -C ₂ -C ₈ -alkenyl, -NHC(O) -C ₂ -C ₈ alkynyl, -NHC(O)-C ₃ -C ₁₂ -cycloalkyl, -NHC(O) aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocycloalkyl, -NHCO ₂ -C ₁ -C ₁₂ - alkyl, -NHCO ₂ -C ₂ -C ₈ -alkenyl, -NHCO ₂ -C ₂ -C ₈ -alkynyl, -NHCO ₂ -C ₃ -C ₁₂ -cycloalkyl, -NHCO ₂ -aryl, -NHCO ₂ -heteroaryl, -NHCO ₂ -heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C ₁ -C ₁₂ -alkyl, -NHC(O)NH-C ₂ -C ₈ -alkenyl, -NHC(O)NH- C ₂ -C ₈ alkynyl, -NHC(O)NH-C ₃ -C ₁₂ -cycloalkyl, -NHC(O)NHaryl, -NHC(O)NH-heteroaryl, -NnC(O)NH-heterocycloalkyl, NHC( S) NH2, -NHC (S) NH-C ₁ -C ₁₂ -alkyl, -NHC (S) NH-C ₂ -C ₈ -alkenyl, -NHC (S) NH-C ₂ -C ₈ -alkynyl, - NHC(S)NH-C ₃ -C ₁₂ -cycloalkyl, -NHC(S)NHaryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH )NH-C ₁ -C ₁₂ -alkyl, -NHC (NH) NH-C ₂ -C ₈ -alkenyl, -NHC (NH) NH-C ₂ -C ₈ -alkynyl, -NHC (NH) NH-C ₃ -C ₁₂ -cycloalkyl, -NHC (NH) NH-aryl, -NHC (NH) NH-heteroaryl, -NHC (NH) NH-heterocycloalkyl, -NHC (NH) -C ₁ -C ₁₂ -al kil, -NnC (NH) -C ₂ -C ₈ -alkenyl, -NHC (NH) -C ₂ -C ₈ -alkynyl, -NHC (NH) -C ₃ -C ₁₂ -cycloalkyl, NnC (NH) aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C ₁ -C ₁₂ -alkyl, -C (NH)NH-C ₂ -C ₈ -alkenyl, -C (NH )NH-C ₂ -C ₈ -alkynyl, -C (NH) NH-C ₃ -C ₁₂ -cycloalkyl, -C (NH) NH-aryl, -C (NH) NH-heteroaryl, -C (NH) NH -heterocycloalkyl, -S (O) -C ₁ -C ₁₂ -alkyl, -S (O) -C ₂ -C ₈ -alkenyl, -S (O) -C ₂ -C ₈ -alkynyl, -S (O) -C ₃ C ₁₂ -cycloalkyl, -S (O) aryl, -S (O) -heteroaryl, -S (O) -heterocycloalkyl, -SO2NH2, -SO ₂ NH-C ₁ -C ₁₂ -alkyl, -SO ₂ NH-C ₂ -C ₈ -alkenyl, -SO ₂ NH-C ₂ -C ₈ -alkynyl, -SO ₂ NH-C ₃ -C ₁₂ -cycloalkyl, -SO ₂ NH-aryl, -SO ₂ NH-heteroaryl, -SO ₂ NH-heterocycloalkyl, -NHSO ₂ -C ₁ -C ₁₂ -alkyl, -NHSO ₂ -C ₂ -C ₈ -alkenyl, -NHSO2-C2-C8 alkynyl, -NHSO ₂ -C ₃ -C ₁₂ -cycloalkyl, -NHSO ₂ -aryl, -NHSO ₂ -heteroaryl, -NHSO ₂ -heterocycloalkyl, -CH2NH2, -CH ₂ SO ₂ CH ₃ , aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, -C ₃ -C ₁₂ cycloalkyl, polyalkoxyalkyl, polyalkoxy , methoxymet hydroxy, methoxyethoxy, -SH, -S-C ₁ -C ₁₂ -alkyl, -SC ₂ -C ₈ -alkenyl, -S-C ₂ -C ₈ -alkynyl, -SC ₃ -C ₁₂ -cycloalkyl, -S- aryl, -S-heteroaryl, -S-heterocycloalkyl or methylthiomethyl. In certain embodiments of the invention, the substituents are independently selected from halogen, preferably Cl and F; N-N-alkyl, preferably methyl and ethyl; C ₂ -C ₄ -alkenyl; halo-C ₁ -C 4 -alkyl, namely fluoromethyl, difluoromethyl and trifluoromethyl; halo-C ₂ -C 4 -alkenyl; C ₃ -C6-cycloalkyl, namely cyclopropyl; -CN; -HE; NH2; C1- _C4 alkylamino; di(C1- _C4 -alkyl)amino and NO2. It is understood that substituents, namely aryls, heteroaryls, alkyls, and the like. are optionally further substituted.

In each case, each substituent on a substituted moiety is additionally optionally substituted with one or more groups, each group being independently selected from C1- _C4 alkyl; CF ₃ , C ₁ -C ₄ alkoxy; -OCF ₃ , -F, -Cl, -Br, -I, -OH, -NO2, -CN and -NH2.

It is understood that aryls, heteroaryls, alkyls, cycloalkyls, and the like. are additionally substituted.

The term halo or halogen, alone or as part of another substituent, as used herein refers to a fluorine, chlorine, bromine or iodine atom.

The term optionally substituted as used herein means that the specified group is substituted or unsubstituted. In one embodiment of the invention, said group is optionally substituted with zero substituents, i. said group is unsubstituted. In another embodiment of the invention, the specified group is optionally substituted with one or more additional(s) group(s), individually and independently selected from the groups described in this document.

The term hydrogen includes hydrogen and deuterium. Additionally, the description of an atom includes other isotopes of that atom, provided that the resulting compound is pharmaceutically acceptable.

In certain embodiments of the invention, it is understood that the compounds of each formula given in this document include isotopically labeled compounds. An isotopically labeled compound is a compound in which at least one atomic site is enriched in a particular isotope of a given element to a level that far exceeds that found in nature. For example, one or more atomic positions in the compound may be enriched in deuterium to a level that is significantly higher than the abundance of deuterium in nature, such as enriched to a level of at least 1%, preferably at least 20%, or preferably at least 50%. Such deuterated compounds may, for example, be metabolized more slowly than their non-deuterated counterparts and therefore exhibit a longer half-life when administered to a subject. Such compounds can be synthesized using techniques known in the art, for example using deuterated starting materials. Unless otherwise stated, isotopically labeled compounds are pharmaceutically acceptable.

The term hydroxyl activating group as used herein refers to a mobile moiety of a chemical molecule known in the art to activate a hydroxyl group such that it will be removed during synthetic processes, namely a substitution or elimination reaction. Examples of the hydroxyl group activating group include, but are not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate, and the like.

The term activated hydroxyl group as used herein refers to a hydroxyl group, an activated group, an activating hydroxyl group as defined above, including, for example, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups.

The term hydroxyl protecting group as used herein refers to a mobile moiety of a chemical molecule known in the art for protecting a hydroxyl group from unwanted reactions during synthetic processes. After said synthetic process(es), the hydroxyl protecting group as described herein is selectively removed. Hydroxyl protecting groups known in the art are generally described in T.N. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, t-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t- butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenylmethyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

The term protected hydroxyl group as used herein refers to a hydroxyl group protected by a hydroxyl protecting group as defined above, including, for example, benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl.

The term inactive hydroxyl group, as used herein, refers to a protecting group known in the art to modify the physicochemical and therefore biological properties of the parent drug in a transient manner by coating or masking the hydroxyl group. Following said synthetic process(es), the inactive hydroxyl group described herein must be capable of being reduced back to a hydroxyl group in vivo. Inactive hydroxyl groups known in the art are described in general terms in Kenneth B. Sloan, Prodrugs, Topical and Ocular Drug Delivery (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York and Prodrugs of Alcohols and Phenols by S.S. Dhareshwar and V.J. Stella, in Prodrugs Challenges and Rewards Part-2 (Biotechnology: Pharmaceutical Aspects), edited by V.J. Stella, et al., Springer and AAPSPress, 2007, pp. 31-99.

The term amino protecting group as used herein refers to a mobile moiety of a chemical molecule known in the art for protecting an amino group from unwanted reactions during synthetic processes. After said process(es) of the synthesis, the amino protecting group as described herein is selectively removed. Amino protecting groups known in the art are described in general terms in T.H. Greene, P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, methoxycarbonyl, t-butoxycarbonyl, (9-fluorenyl)methoxycarbonyl, benzyloxycarbonyl, and the like.

The term protected amino group as used herein refers to an amino group protected by an amino group protecting group as defined above.

The term amino acid refers to naturally occurring and synthesized α, β, γ, or δ-amino acids and includes, but is not limited to, amino acids found in proteins and intermediates in amino acid or protein metabolism, i. glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, aspargine, glutamine, aspartate, glutamate, lysine, citrulline, arginine, and histidine. In certain embodiments of the invention, the amino acid is in the L-configuration. In certain embodiments of the invention, the amino acid is in the D-configuration. In certain embodiments, amino acids are provided as substitutes for a compound described herein, wherein the amino acid is a residue selected from the group consisting of alanyl, valinyl, leucinyl, isoleucinyl, prolinyl, phenylalaninyl, tryptophanyl, methioninyl, glycinyl, serinyl, threoninyl , cysteinyl, tyrosinyl, asparaginyl, glutaminyl, aspartoyl, glutaroyl, lysinyl, argininyl, histidinyl, β-alanyl, β-valinyl, β-leucinyl, β-isoleucinyl, β-prolinyl, β-phenylalaninyl, βtryptophanyl, β-methioninyl, β β-glycinyl, β-serinyl, β-threoninyl, β-cysteinyl, β-tyrosinyl, β-asparaginyl, β-glutaminyl, β-aspartoyl, β-glutaroyl, β-lysinyl, β-argininyl and β-histidinyl.

The term amino acid derivative refers to a group derived from a naturally occurring or non-naturally occurring amino acid described and exemplified herein. Amino acid derivatives are obvious to those skilled in the art and include, but are not limited to, esters, amino alcohols, amino aldehydes, amino lactones, and N-methyl derivatives of naturally occurring or non-naturally occurring amino acids. In one embodiment, an amino acid derivative is provided as a substituent on a compound described herein, wherein the substituent is -NR ^u -G(Sc)-C(O)-Q1, where Q ¹ is -SR ^v , -NR ^v R ^v or alkoxy, R ^v is hydrogen or alkyl, S _c is a naturally occurring or non-naturally occurring side chain

- 18 041670 the nature of the amino acid, G is C1-C2 alkyl and R ^u is hydrogen; or R ^u and S _c together with the atoms to which they are attached form a five-membered heterocyclic ring. In one embodiment, an amino acid derivative is provided as a substituent on a compound described herein, wherein the substituent is -OC(O)-G(S _c )NH-Q2, where Q2 is hydrogen or alkoxy, S _c is a naturally occurring or non-naturally occurring amino acid side chain; and G is C1-C2 alkyl. In certain embodiments of the invention, Q2 and S _c, together with the atoms to which they are attached, form a five-membered heterocyclic ring. In certain embodiments, G is an optionally substituted methylene and Sc is selected from the group consisting of hydrogen, alkyl, arylalkyl, heterocycloalkyl, carboxyalkyl, heteroarylalkyl, aminoalkyl, hydroxyalkyl, aminoiminoaminoalkyl, aminocarbonylalkyl, sulfanylalkyl, carbamoylalkyl, alkylsulfanylalkyl, and hydroxyarylalkyl. In one embodiment, an amino acid derivative is provided as a substituent on a compound described herein, wherein the amino acid derivative is in the D configuration. In one embodiment, an amino acid derivative is provided as a substituent on a compound described herein, wherein the amino acid derivative is in the L-configuration.

The term substitutable group means a functional group or atom that can be replaced by another functional group or atom in a substitution reaction, namely a nucleophilic substitution reaction. By way of example, typical substitutable groups include groups such as chlorine, bromine, and iodine, groups such as sulfonic esters, namely mesylate, tosylate, brosylate, nosylate, and the like, and groups such as alkoxy, namely acetoxy. , trifluoroacetoxy, and the like.

The term aprotic solvent as used herein refers to a solvent that is relatively inert to protic activity, i. which does not act as a proton donor. Examples include, but are not limited to, hydrocarbons, namely, for example, hexane and toluene, halogenated hydrocarbons, namely, for example, methylene chloride, ethylene chloride, chloroform, and the like, heterocyclic compounds, namely, for example, tetrahydrofuran and N- methylpyrrolidinone, and ethers, namely diethyl ether, bis-methoxymethyl ether. Such compounds are well known to those skilled in the art, and it will be apparent to those skilled in the art that individual solvents or mixtures thereof are preferred for certain compounds and reaction conditions, depending on such factors as, for example, the solubility of the reagents, the reactivity of the reagents and preferred temperature ranges. Additional descriptions of aprotic solvents can be found in textbooks of organic chemistry or in specialized monographs such as Organic Solvents Physical Properties and Methods of Purification, 4 ^th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The term protic solvent as used herein refers to a solvent capable of producing protons, namely an alcohol such as methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and it will be apparent to those skilled in the art that individual solvents or mixtures thereof are preferred for certain compounds and reaction conditions, depending on such factors as, for example, the solubility of the reagents, the reactivity of the reagents and preferred temperature ranges. Additional descriptions of protogenic solvents can be found in textbooks of organic chemistry or in specialized monographs such as Organic Solvents Physical Properties and Methods of Purification, ^4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The combinations of substituents and variables contemplated by this invention are only those that form stable compounds. The term stable, as used herein, refers to compounds that are stable enough for manufacture, and that retain the integrity of the compound for a sufficient period of time to be used for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

The synthesized compounds are separated from the reaction mixture and further purified by methods such as column chromatography, high pressure liquid chromatography or recrystallization. Those skilled in the art will recognize that those skilled in the art will recognize additional methods for synthesizing compounds of the formula provided herein. Additionally, the various steps of the synthesis can be performed in an alternative sequence or order to obtain the desired compounds. The chemical transformations during synthesis and the methodology of protecting groups (protection and deprotection) useful in the synthesis of the compounds described herein are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, 2 ^n.d. Ed. Wiley-VCH (1999); TW Greene, PGM Wuts, Protective Groups in Organic Synthesis, ^3rd Ed., John Wiley and Sons (1999); L. Fieser, M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994) and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and their subsequent editions .

- 19 041670

The term subject as used herein refers to an animal. Preferably the animal is a mammal. More preferably, the mammal is a human. The subject also refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, fish, birds, and the like.

The compounds of this invention can be modified by adding suitable functional groups to improve the selective biological properties. Such modifications are known in the art and may include those that increase biopermeability in a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase injectable solubility, alter metabolism, and alter excretion rate.

The compounds described herein contain one or more asymmetric centers and therefore form enantiomers, diastereomers, and other stereoisomeric forms defined in terms of absolute stereochemistry as (R)- or (S)-, or as (D)- or ( L) - for amino acids. This invention is intended to include all such possible isomers as well as their racemic and optically pure forms. Optical isomers are obtained from their respective optically active starting compounds by the methods described above, or by separation of racemic mixtures. Separation is carried out in the presence of a resolving agent, chromatography or recrystallization, or some combination of these methods known to those skilled in the art. Additional details regarding separation can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When the compounds described herein contain an olefinic double bond, other regions of unsaturation, or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are intended to include both E and Z geometric isomers, or cis and trans isomers. Likewise, all tautomeric forms are also intended to be included. Tautomers may be cyclic or acyclic. The configuration of any carbon-carbon double bond shown herein is for convenience only and is not intended to be a specific configuration unless otherwise indicated in the text; therefore, a carbon-carbon double bond or a carbon-heteroatomic double bond, arbitrarily depicted herein as trans, is cis, trans, or a mixture of both in various ratios.

Certain compounds of this invention also exist in different stable conformational forms that can be separated. Torsion asymmetry due to limited rotation around the asymmetric single bond, for example due to steric hindrance or ring deformation, allows separation of different conformers. This invention includes each conformational isomer of these compounds or mixtures thereof.

As used herein, the term pharmaceutically acceptable salt refers to those salts that are medically suitable for contact with human and lower animal tissues without causing undue intoxication, irritation, allergic reaction, etc., and are commensurate with the therapeutic index. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge, et al., describe in detail pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts are obtained in situ during the final isolation or purification of the compounds of the invention, or separately by reaction of the free base with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to: non-toxic acid addition salts are salts of the amino group attached to an inorganic acid, namely hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or organic acids, and specifically with acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or using other methods known in the art, namely ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, salts such as adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate , glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodine, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate , pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Typical alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Additional pharmaceutically acceptable salts include, if necessary, non-toxic ammonium, quaternary ammonium and amine cations formed with counterions, namely halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate .

As used herein, the term pharmaceutically acceptable ester refers to esters that are hydrolyzed in vivo and include those that are readily degraded in the human body to form the parent compound or a salt thereof. Suitable esters

- 20 041670 groups include, for example, derivatives of pharmaceutically acceptable aliphatic carboxylic acids, namely alkanoic, alkenoic, cycloalkanoic and alkenedioic acids, in which each alkyl or alkenyl fragment preferably contains no more than 6 carbon atoms.

Examples of specific esters include, but are not limited to, esters of C1-C6 alkanoic acids, namely acetate, propionate, butyrate and pivalate ester.

pharmaceutical compositions.

Pharmaceutical compositions of this invention contain a pharmaceutically effective amount of a compound of this invention in combination with one or more pharmaceutically acceptable carriers or excipients.

As used herein, the term pharmaceutically acceptable carrier or excipient means a non-toxic, inert solid, soft, or liquid excipient, diluent, encapsulating material, or excipient of any kind. Some examples of materials that serve as pharmaceutically acceptable carriers are sugars, namely lactose, glucose and sucrose; starches, namely corn starch and potato starch; cellulose and its derivatives, namely sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powder tragacanth; malt; gelatin; talc; excipients, namely cocoa butter and suppository waxes; oils, namely peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, namely propylene glycol; esters, namely ethyl oleate and ethyl laurate; agar; buffer substances, namely magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic solution; Ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other non-toxic compatible lubricants, namely sodium lauryl sulfate and magnesium stearate, as well as colorants, anti-adherents, coating agents, sweeteners, flavors and flavors, preservatives and antioxidants are also in the composition, according to pharmacist's point of view.

Pharmaceutical compositions of this invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, preferably by oral or injection route. The pharmaceutical compositions of this invention contain any conventional non-toxic pharmaceutically acceptable carriers, excipients or excipients. In some cases, the pH of the composition is adjusted with pharmaceutically acceptable acids, bases, or buffers to improve the stability of the drug compound or its delivery form. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, introsynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion route of administration.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and infusions. In addition to the active compounds, liquid dosage forms contain inert diluents widely used in the art, namely, for example, water or other solvents, solubilizing agents and emulsifiers, namely ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (particularly cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, compositions for oral administration also include excipients, namely wetting agents, emulsifying agents and suspension stabilizers, sweeteners, flavors and perfumes.

Injectable formulations, for example sterile injectable aqueous or oily suspensions, are prepared according to methods known in the art using suitable dispersing or wetting agents and suspension stabilizers. A sterile injectable preparation is also a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable carriers or solvents that may be used are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. Additionally, sterile fixed oils have traditionally been used as the solvent or dispersion medium. For this purpose, any pure fixed oil is used, including synthetic mono- or diglycerides. Additionally, fatty acids, namely oleic acid, are used in the preparation of injectables.

Injectable formulations are sterilized, for example, by filtration through a bacteria-retaining filter, or by administering sterilizing agents in the form of sterile solid formulations that are dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of a drug, it is often necessary to slow down the absorption of the drug from subcutaneous or intramuscular injection. This is achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility. In this case, the rate of adsorption of the drug depends on the rate of its dissolution, which, in turn, depends on the size of the crystal and the crystalline form. Alternatively, delayed absorption of a parenterally administered dosage form is achieved by dissolving or dispersing the drug in an oil vehicle. Sustained absorption injectable formulations are prepared by incorporating microcapsular matrices of the drug into biodegradable polymers, namely polyactide-polyglycolide. The rate of drug release is controlled depending on the ratio of drug to polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Delayed injectable formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories which are prepared by mixing the compounds of this invention with suitable non-irritating excipients and carriers, namely cocoa butter, polyethylene glycol or suppository wax, which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active ingredient is admixed with at least one inert pharmaceutically acceptable excipient or carrier, namely sodium citrate or dicalcium phosphate, and/or: a) excipients or diluents, namely starches, lactose, sucrose, glucose, mannitol and silicic acid; b) binders, namely, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and gum; c) wetting agents, namely glycerin; d) disintegrators, namely agaragar, calcium carbonate, potato and tapioca starch, alginic acid, certain silicates and sodium carbonate; e) substances that slow down the dissolution, namely paraffin; f) absorption accelerators, namely quaternary ammonium compounds; g) substances that improve wettability, namely, for example, cetyl alcohol and glyceryl monostearate; h) absorbents, namely kaolin and bentonite clay; and i) lubricants, namely talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form also contains buffering agents.

Solid compositions of a similar type are also used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

Solid dosage forms of tablets, dragees, capsules, pills and granules are prepared with coatings or shells, namely enteric coatings or other coatings well known in the art of drug preparation. They optionally contain opacifiers and are formulated to release the active ingredient(s) only, or preferentially, in a specific part of the intestine, optionally in a delayed manner. Examples of useful encapsulating compositions include polymeric materials and waxes.

Dosage forms for surface or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, volatile preparations, and patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or required buffers. It is also intended that the scope of the invention include ophthalmic preparation, ear drops, eye ointments, powders and solutions.

Ointments, pastes, creams and gels contain, in addition to the active compound according to the invention, excipients, namely animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

The powders and sprays contain, in addition to the compounds according to the invention, excipients, namely lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and powdered polyamide, or mixtures of these substances. Sprays additionally contain traditional spray substances, namely chlorofluorocarbons.

Transdermal patches have the added benefit of providing controlled delivery of the compound to the body. Such dosage forms are prepared by dissolving or distributing the compound in the proper medium. Absorption enhancers are also used to increase the flux of the compound through the skin. The rate is controlled either by using a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

For inhalation delivery, the therapeutic composition of this invention is formulated and administered to a patient in solid or liquid aerosol form by direct administration, eg, inhalation into the respiratory system. Solid or liquid aerosol forms of the active compound for carrying out this invention contain particles of respirable size, i. particles are small enough

- 22 041670 size to pass through the mouth and larynx when inhaled and enter the bronchi and alveoli of the lungs. Delivery of aerosol therapeutic agents, in particular aerosol antibiotics, is known in the art (see, for example, US Pat. No. 5767068, Van Devanter et al., US Pat. No. 5508269, Smith et al. and WO 98/43650, Montgomery , all of which are incorporated herein by reference).

antiviral activity.

The inhibitory amount or dose of the compounds of this invention ranges from about 0.01 to about 500 mg/kg, alternatively from about 1 to about 50 mg/kg. Inhibitory amounts or doses also depend on the route of administration, as well as on the possibility of co-administration with other agents.

According to the methods of treatment of this invention, viral infections and diseases are treated or prevented in patients such as humans or other animals by administering to the patient a therapeutically effective amount of a compound of the invention, in such amounts and for such time as is necessary to achieve the desired result.

By a therapeutically effective amount of a compound of the invention is meant an amount of a compound that has a therapeutic effect on the subject being treated, at an acceptable therapeutic index applicable to any medical treatment. The therapeutic effect is objective (ie measurable by some test or marker) or subjective (ie the subject notices or feels the effect). An effective amount of the compound described above is in the range of about 0.1 to about 500 mg/kg, preferably about 1 to about 50 mg/kg. Effective doses also depend on the route of administration, as well as on the possibility of co-administration with other agents. It is understood, however, that the total daily dose of the compounds or compositions of this invention is administered after a visit to a physician within the scope of a thorough medical evaluation. The exact therapeutically effective dose level for any given patient depends on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the particular compound used; the particular composition used; age, body weight, general health, sex and nutrition of the patient; the time of administration, route of administration, and rate of excretion of the particular compound used; duration of treatment; drugs used in combination or simultaneously with the particular compound used; and similar factors well known in the medical arts.

The total daily dose of the compounds of this invention, administered to a human or other animal, in single or divided doses, is in the range, for example, from 0.01 to 50 mg/kg of body weight, or more commonly from 0.1 to 25 mg/kg of body weight. body. A unit dose of the compositions contains such amounts or proportions thereof that constitute a daily dose. Typically, the treatment regimen of this invention comprises administering to a patient in need of such treatment from about 10 to about 1000 mg of the compound(s) of this invention per day in single or divided doses.

The compounds of the invention described herein are administered, for example, by injection, intravenously, intra-arterially, subcutaneously, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in an ophthalmic formulation, or by inhalation, at a dosage in the range of about 0.1 to about 500 mg/kg body weight, alternatively, at a dosage of 1 mg to 1000 mg/kg dose, every 4 to 120 hours, or as required for a particular medicinal product. The methods herein involve administering an effective amount of a compound or a composition with a compound to achieve a desired or permanent effect. Typically, the pharmaceutical compositions of this invention are administered from about 1 to about 6 times per day, or alternatively, as a continuous infusion. Such an introduction is used as a long-term or intensive therapy. The amount of active ingredient that is combined with pharmaceutical excipients or carriers to form a unit dosage form depends on the recipient being treated and the particular route of administration. A typical formulation contains from about 5% to about 95% active compound (w/w). Alternatively, such formulations contain from about 20% to about 80% active compound.

Lower or higher doses than those listed above may be required. The specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the potency of the particular compound employed, age, body weight, general health, sex, diet, time of administration, rate of excretion, combination of drugs, severity and course of illness, disease or symptoms, the patient's susceptibility to the disease, disorder or symptoms, and evaluation by the attending physician.

When the patient's condition improves, if necessary, a maintenance dose of a compound, composition or combination of this invention is administered. Therefore, the dosage or frequency of administration, or both, is reduced, as a function of symptoms, to a level at which improved condition is maintained when symptoms are reduced to the target level. Patients, however, may require intermittent therapy on a long-term basis for any recurrence of disease symptoms.

- 23 041670

In case the compositions of this invention contain a combination of a compound of the formula described herein and one or more additional therapeutic or prophylactic agents, and the compound and additional agent should be in a dosage of from about 1 to 100% and preferably from about 5 to 95 % of the dose usually given as monotherapy. Additional funds are administered separately, as part of multiple doses, of the compounds of this invention. Alternatively, these agents are part of a unit dosage form mixed together with the compounds of this invention into a single composition.

These additional therapeutic or prophylactic agents include, but are not limited to, immunotherapy agents (eg, interferon), therapeutic vaccines, antifibrotic agents, anti-inflammatory agents, namely corticosteroids or NSAIDs, bronchodilators, namely beta-2-adrenergic agonists, and xanthines (eg. , theophylline), mucolytics, antimuscarinics, leukotriene receptor inhibitors, non-specific cell adhesion inhibitors (eg, ICAM antagonists), antioxidants (eg, Nacetylcysteine), cytokine agonists, cytokine antagonists, pulmonary surfactants, and/or antimicrobial and antiviral agents (eg, ribavirin and amantidine). The compositions of the invention are also used in combination with gene replacement therapy. Combination or alternative therapy for HBV

It was found that after long-term treatment with an antiviral agent, drug-resistant variants of HIV, HBV, HCV appear. Drug resistance most often results from a mutation in a gene that codes for a protein, namely an enzyme used in viral replication, and most often in the case of HIV reverse transcriptase, protease or DNA polymerase, and in the case of HBV, DNA polymerase, or in in the case of HCV, RNA polymerase, protease or helicase. It has recently been shown that the efficacy of an HIV drug can be prolonged, improved or restored by administering the compound in combination or modification with a second and possibly third antiviral compound that causes a mutation other than that caused by the parent drug. The compounds are used for combination and may be selected from the group consisting of an HBV polymerase inhibitor, interferon, TLR modulators, namely TLR-7 agonists or TLR-9 agonists, therapeutic vaccines, immunoactivators of certain cellular viral RNA sensors, virus entry inhibitor, maturation inhibitor a virus, a certain capsid assembly modulator, antiviral compounds of a certain or unknown mechanism, and combinations thereof. Alternatively, the pharmacokinetics, biodistribution, and other parameters of the drug are altered by such a combination or alternative therapy. In general, combination therapy is generally preferred over alternative therapies because it creates multiple simultaneous exposures to the virus.

Preferred compounds for combination or alternative therapy for the treatment of HBV include 3TC, FTC, L-FMAU, interferon, adefovir dipivoxil, entecavir, telbivudine (L-dT), valtorcitabine (3'-valinyl L-dC), e-D-dioxolanylguanine (DXG) , β-O-dioxolanyl-2,6-diaminopurine (DAPD) and β-O-dioxolanyl-6-chloropurine (ACP), famciclovir, penciclovir, lobukavir, ganciclovir and ribavirin.

Although the invention has been described in accordance with various preferred embodiments, the invention is not intended to be limited thereto, but rather, those skilled in the art will appreciate that changes or modifications may be made to this document which are within the spirit of the invention and the scope of the appended claims. inventions. Abbreviations

The abbreviations used in the descriptions of circuits and examples are as follows:

Ac - acetyl;

AcOH - acetic acid;

AIBN - azobisisobutyronitrile;

BINAP - 2,2'-bis-(diphenylphosphino)-1,1'-binaphthyl;

Boc ₂ O - di-tert-butyl dicarbonate;

Boc - t-butoxycarbonyl;

Bros - 1-methyl-1-(4-biphenylyl)ethylcarbonyl;

Bz - benzoyl;

Bn - benzyl;

BocNHOH - tert-butyl-N-hydroxycarbamate;

t-BuOK - potassium tert-butoxide;

Bu ₃ SnH - tributyltin hydride;

BOP - (benzotriazol-1-yloxy)-tris-(dimethylamino)phosphonium hexafluorophosphate;

brine - a solution of sodium chloride in water;

BSA - N,O-bis-(trimethylsilyl)acetamide;

- 24 041670

CDI - carbonyldiimidazole;

DCM or CH2C12 - dichloromethane;

CH ₃ - methyl;

CH ₃ CN - acetonitrile;

Cs ₂ CO ₃ - cesium carbonate;

CuCl - copper(1) chloride;

CuI - copper(I) iodide;

dba - dibenzylideneacetone;

dffb - diphenylphosphinobutane;

DBU - 1,8-diazobicyclo[5.4.0]undec-7-ene;

DCC - N,N'-dicyclohexylcarbodiimide;

DEAD - diethyl azodicarboxylate;

DIAD - diisopropyl azodicarboxylate;

DIPEA or (i-Pr)2EtN - N,N,-diisopropylethylamine;

Dess-Martin periodate - 1,1,1-tris-(acetyloxy)-1,1-dihydro-1,2-benzyodoxol-3-( 1 H) -one;

DMAP - 4-dimethylaminopyridine;

DME - 1,2-dimethoxyethane;

DMF - N,N-dimethylformamide;

DMSO - dimethyl sulfoxide;

DMT - di-(p-methoxyphenyl)phenylmethyl or dimethoxytrityl;

DPFA - diphenylphosphoryl azide;

EDC - N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide;

EDC HCl - N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride;

EtOAc-ethyl acetate;

EtOH - ethanol;

Et ₂ O - diethyl ether;

HATU - O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

HCl - hydrogen sulfide;

HOBT - 1-hydroxybenzotriazole;

K ₂ CO ₃ - potassium carbonate;

n-BuLi - n-butyllithium;

i-BuLi - i-butyllithium;

t-BuLi - t-butyllithium;

PhLi - phenyllithium;

LDA - lithium diisopropylamide;

LiTMP - lithium 2,2,6,6-tetramethylpiperidine;

MeOH - methanol;

Mg - magnesium;

MOM - methoxymethyl;

Ms is mesyl- or -SO2-CH3;

Ms ₂ O is methanesulfonic anhydride or mesylanhydride;

MTBE - t-butyl methyl ether;

NaN(TMS) ₂ - sodium bis(trimethylsilyl)amide;

NaCl - sodium chloride;

NaH - sodium hydride;

NaHCO3 - sodium bicarbonate or sodium hydrogen carbonate;

Na2CO3 - sodium carbonate;

NaOH - sodium hydroxide;

Na ₂ SO ₄ - sodium sulfate;

NaHSO ₃ - sodium bisulfite or sodium acid sulfite;

Na ₂ S ₂ O ₃ - sodium thiosulfate;

NH ₂ NH ₂ - hydrazine;

NH ₄ HCO ₃ - ammonium bicarbonate;

NH ₄ Cl - ammonium chloride;

NMO - N-methylmorpholine-N-oxide;

NaIO4 - sodium periodate;

Ni - nickel;

NSFI - N-fluorobenzenesulfonimide;

OH - hydroxyl group;

OsO4 - osmium tetroxide;

RTSA - p-toluenesulfonic acid;

RPTS - pyridinium p-toluenesulfonate;

- 25 041670

TBAF - tetrabutylammonium fluoride;

TEA or Et ₃ N - triethylamine;

TES - triethylsilyl;

TES Cl - triethylsilyl chloride;

TESOTf - triethylsilyl trifluoromethanesulfonate;

TFA - trifluoroacetic acid;

THF - tetrahydrofuran;

TMEEDA - N,N,N',N'-tetramethylethylenediamine;

TFF or PPh ₃ - triphenylphosphine;

Troc - 2,2,2-trichloroethylcarbonyl;

Ts - tosyl or -SO ₂ -C ₆ H ₄ CH ₃ ;

Ts ₂ O - tolylsulfonic anhydride or tosyl anhydride;

TsOH - p-tolylsulfonic acid;

Pd - palladium;

Ph - phenyl;

POPd dihydrogendichlorobis-(di-tert-butylphosphinito-cP)palladate(II);

Pd ₂ (dba) ₃ - dipalladium(0) tris-(dibenzylideneacetone);

Pd(PPh ₃ ) ₄ - palladium(0) tetrakis-(triphenylphosphine);

PdCl ₂ (PPh ₃ ) ₂ - palladium(II) trans-dichlorobis-(triphenylphosphine);

Pt - platinum;

Rh - rhodium;

kt - room temperature;

Ru - ruthenium;

SLC - supercritical liquid chromatography;

TBS - tert-butyldimethylsilyl;

TMS - trimethylsilyl or

TMS Cl - trimethylsilyl chloride.

Synthesis methods.

The compounds and processes of this invention will be better understood in conjunction with the following synthetic schemes, which illustrate the methods by which the compounds of the invention are prepared. These diagrams are for illustrative purposes and are not intended to limit the scope of the invention. Equivalent, similar, or acceptable solvents, reagents, and reaction conditions may be substituted for the specific solvents, reagents, or reaction conditions described herein without deviating from the overall scope of the synthesis process. Certain reactions may be carried out as generally described in WO 2017/136403.

The compounds of formula I can be obtained through several different synthetic routes from a variety of optionally substituted phenyl, heteroaryl, or fused bicyclic aryl or heteroaryl starting materials using chemical transformations known to those skilled in the art. From a strategic point of view, the compound of formula I can be created with the formation of the sulfonyl group at the right end and the subsequent formation of group A at the left end. Alternatively, a compound of formula I can be designed to form an A group at the right end followed by a sulfonyl group at the left end. The production of sulfones can be realized either by sulfide oxidation (reviewed by K. Schank, The Chemistry of Sulfones and Sulfoxides, Wiley, New York, 1988, Chap.7), or by alkylation/arylation of low-valence sulfur compounds, namely salts such as sulfinates (reviewed G. Liu, C. Fan, J. Wu, Org Biomol Chem 2015, 13, 1592). The sulfide can be synthesized from a thiol starting material by nucleophilic substitution of an organic halide or sulfonic acid ester, or nucleophilic addition to an epoxide, arizidine, or unsaturated base (reviewed by G. Solladie, Comprehensive Organic Synthesis, 1991, Vol. 6, 133), or radical addition of a thiol to an unsaturated base. A salt such as a sulfinate can be prepared either by reduction of a sulfonyl halide (reviewed by Schubart, R. Sulfinic Acids and Derivatives, Ullmann's Encyclopedia of Industrial Chemistry, 2000, 677) or by a transition metal catalyzed reaction with an aryl or heteroaryl halide (A. Shavnya, S.S. Coffey, A. C. Smith, V. Mascitti, Org. Lett., 2013, 15, 6226) or boric acid (A. Shavnya, K. D. Hesp, V. Mascitti, A. C. Smith, Angew. Chem. Int. Ed. , 2015, 54, 13571) with potassium metabisulfite. To a compound such as a sulfone, functional groups can be further added by deprotonization with a strong base, followed by reaction of the resulting anion with an electrophilic compound, namely an organic halide, an aldehyde, a ketone, an electrophilic halogenation reagent, or an unsaturated base, namely an additive Michael acceptor; a quaternary sulfone can be prepared from a primary sulfone by a two step sequential deprotonization and an anionic nucleophilic reaction. The amide bond can be prepared either by reacting an acid halide or anhydride with an amine, or by directly coupling a carboxylic acid with an amine in the presence of a coupling reagent, namely DCC,

- 26 041670

EDC or HATU.

As depicted in Scheme 1, where X, Y and R have previously been described, LG1, LG2 are in each case substitutable groups and are each independently halogen, tosylate, mesylate and triflate. In one approach, an optionally substituted aryl- or heteroarylamine 1-1 can selectively react with various acid chlorides 1-2 in a solvent, namely, but not limited to, toluene, tetrahydrofuran, dichloromethane, or mixtures thereof, optionally in the presence of a base, namely, but not limited to, triethylamine, DIPEA or pyridine, to obtain a variety of intermediate amides 1-3. 1-3 is then treated with a reducing agent, namely, but not limited to, triphenylphosphine, SnCl ₂ , Sn/HCl, Zn/HCl, or Pd/HCOOH, to give thiol intermediates 1-4 which react with intermediate 1-5 in order of nucleophilic substitution, optionally in the presence of a base, namely, but not limited to, potassium carbonate, sodium carbonate, triethylamine or DIPEA, to obtain a sulfide intermediate, which is converted to a compound of formula IIa in a suitable solvent in the presence of an oxidizing agent, namely , but not limited to, hydrogen peroxide, metachlorobenzoic acid, perbenzoic acid, or tert-butyl peroxide. Alternatively, the carboxylic acid ester 1-6 is converted to the sulfonic intermediate 1-7 using chemistry similar to that described above or organometallic nucleophilic substitution (RM where M is Mg or Zn). 1-7 can be partially saponified with a base, namely, but not limited to, lithium hydroxide, sodium hydroxide or potassium hydroxide, to obtain carboxylic acid 1-8. Acid 1-8 may be reacted with amine 1-1 in the presence of a coupling reagent, namely, but not limited to, DCC, EDC, or HATU, in a suitable solvent, optionally in the presence of a base, namely, but not limited to, triethylamine, DIPEA or pyridine, to obtain a compound of formula IIa.

Scheme 1

The preparation of the compound of formula IIb is described in Scheme 2 below. Aldehyde 2-1 is reacted with TMSCF3 to form trifluoroethyl alcohol 2-2, which is converted to triflate by reaction with Tf ₂ O in the presence of a base, namely DIPEA, followed by replacement with amine X- NH2 to obtain a compound of formula IIb.

Scheme 2

O O ™SCF ₃ F ₃ C O 1.SchO FjC O

JYSR ---VY-SR --► \_ _y _ _s _r

N y BUT ο 2, * ΝΗ _± Χ-ΝΗ o

2-1 2-2 Hb

The synthesis of a compound of formula IIc containing an aminooxetanyl fragment of the molecule is shown in scheme 3. Arylamine or heteroarylamine 3-1 is condensed with oxetan-3-one in the presence of an acid, namely acetic acid or p-TsA, to obtain imine 3-2, which is treated with a nucleophilic compound 3-3, in which M1 represents organometallic compounds, including, but not limited to, those related to boric acid/boric acid ester, organotin, organozinc, organolithium or organomagnesium fragment of the molecule to obtain compounds of formula IIc.

Scheme 3

As shown in Scheme 4, a compound of formula IId can be prepared from a compound of formula Ha. Na can react with benzyl bromide in the presence of a base, namely, but not limited to, NaH or LDA, to form compound 4-1, which is converted to compound 4-2 by reaction with oxalyl chloride, followed by treatment with a fluorinating reagent, namely, but not limited to DAST, SF4 or Et ₃ N-HF. Compound 4-2 can be treated with a gas such as hydrogen in the presence of a suitable catalyst, namely, but not limited to, Pd/C, PtO ₂ or Pd(OH) ₂ /C, to obtain a compound of formula IId.

- 27 041670

Scheme 4

It will be understood that by suitable means and protection of any chemical functional group, the synthesis of compounds of formula I is carried out by methods analogous to those given above and described in the Experimental Section. Suitable functional groups can be found, but are not limited to, in T.W. Greene and P.G.M. Wuts Protective Groups in Organic Synthesis, 3rd Ed (1999), J Wiley and Sons.

All references listed in this document, whether in print, electronic form, computer media or otherwise, are purposefully referenced in their entirety in this document, including, but not limited to, abstracts, articles, journals, publications, texts, treatises. , internet websites, databases, patents and patent applications.

Various changes and modifications with respect to those disclosed in the embodiments of the invention are obvious to those skilled in the art, and such changes and modifications, including, without limitation, relating to chemical structures, substituents, derivatives, formulations and/or methods of the invention, can be to make without deviating from the essence of the invention and the scope of the attached claims.

While the invention has been described in accordance with various preferred embodiments, the invention is not intended to be limited thereto, but rather, those skilled in the art will appreciate that changes or modifications may be made to this document which are within the spirit of the invention and the scope of the appended claims. inventions.

Examples

The compounds and processes of this invention will be better understood in conjunction with the following examples, which are intended to be illustrative only and do not limit the scope of the invention. Various changes and modifications with respect to those disclosed in the embodiments of the invention are obvious to those skilled in the art, and such changes and modifications, including, without limitation, relating to chemical structures, substituents, derivatives, formulations and/or methods of the invention, can be to make without deviating from the essence of the invention and the scope of the attached claims.

Intermediate 1

Stage prom. 1a. A mixture of 4-chloro-3-(chlorosulfonyl)benzoic acid (0.86 g, 3.4 mmol) in SOCl ₂ (5.0 ml) was heated at reflux overnight. It is concentrated to obtain the desired crude product, which is immediately used in the next step.

Stage prom. 1b. The compound of step 1a (0.91 g, 3.3 mmol) and 3,4,5-trifluoroaniline (0.49 g, 3.3 mmol) in toluene (10 ml) was stirred at 90° C. overnight. The mixture is concentrated to obtain the target crude compound, which is immediately used in the next step.

Stage prom. 1s. The compound of step 1b (0.89 g, 2.3 mmol) and triphenylphosphine (3.4 g, 13 mmol) in toluene (12 ml) were stirred at 80° C. for 4 h. The mixture was diluted with EtOAc and washed with saturated aqueous NaHCO ₃ and brine. The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was chromatographed (silica, hexanes/EtOAc) to give the title compound as a white solid (0.49 g, 71%). ESI-MS m/z=316.0, 318.0 [MH] ^- .

Intermediate 2

Stage prom. 2a. A mixture of 2-methylenepropane-1,3-diyldiacetate (2.69 g, 15.62 mmol), Pd(OAc) ₂ (0.210 g, 0.937 mmol), Ph ₃ P (0.983 g, 3.75 mmol) and 1- (cyclopent-1-en-1-yl)pyrrolidine (3.19 ml, 21.86 mmol) in acetonitrile (89 ml) is heated and maintained at 65° C. for 18 h. Water (45 ml) is added and the reaction the mixture is stirred for 1 hour. Saturated brine is added and the mixture is extracted with ethyl acetate. The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was chromatographed (silica, hexanes/EtOAc) to give the title compound (1.53 g, 71.9% yield) as a colorless oil.

Stage prom. 2b. A solution of the compound of step 2a (41.725 g, 306 mmol) in THF (300 ml) was cooled to 78° C. followed by the addition of LiAlH ₄ (1 M in THF, 92 ml, 92 mmol). After stirring for 15 minutes, the mixture was quenched with water (3.4 ml), NaOH (1 M, 3.4 ml) and water (10.2 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered over Celite and concentrated to give the crude title compound (48.4 g, 97%, containing 15 w/w% THF) which was used in the next step.

Stage prom. 2s. To a stirred compound of step 2b (50.7 g, 367 mmol) and imidazole (62.4 g, 58.8 mmol) in DMF (400 ml) at 0°C was added TBS Cl (66.3 g, 440 mmol). The resulting reaction mixture was stirred at rt for 16 h. The reaction mixture was diluted with hexanes and the mixture was washed with water and brine. The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound as a white solid (101.0 g, 100%).

Stage prom. 2d. To a suspension of the compound of step 2c (101 g, 368 mmol) in dioxane-water (1.1 L/0.36 L) at rt was added 2,6-dimethylpyridine (86 mL, 735 mmol), osmium(VIII) oxide (1 .87 g, 7.35 mmol) and sodium periodate (280 g, 1.31 mol) and the mixture was stirred at rt for 20 h. The mixture was quenched with aqueous Na ₂ S ₂ O ₃ , extracted with MBTE, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the desired product as a white solid (100 g, 99%).

Stage prom. 2e. To a solution of the compound of step 2d (118.5 g, 466 mmol) and MTBE (1.2 L) at 0°C was added LiBH4 (314 mL, 629 mmol, 2 M in THF). The resulting reaction mixture was stirred at 0° C. for 2 hours. The reaction mixture was quenched with aqueous NH4Cl and the mixture was extracted with MBTE, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was used without further purification (117 g, 98%, 10:1 tf (desired ratio, dr, English: desired ratio), contributing to the desired isomer).

Stage prom. 2f To a stirred solution of step 2e (315 mg, 1.23 mmol) and compound of step 1c (390 mg, 1.23 mmol) in toluene (5 ml) was added 2-(tributyl-15-phosphanylidene)acetonitrile (0.81 ml , 3.07 mmol) and the mixture was stirred at 100°C for 60 h. Cooled to rt, diluted with MBTE, washed with NaOH (0.5 N) and brine, dried over Na2SO4, filtered, concentrated, using a column of dioxide silicon to obtain the target compound (362 mg, 53%), ESI-MS m/z=554.15, 556.15 [MH]-.

Stage prom. 2g. Conc. HCl (1.0 ml) and stirred at rt for 24 h. It was concentrated in vacuo to remove most of the MeOH and the residue was extracted with EtOAc. The organic phase was washed with water, 10% K ₂ CO ₃ and brine, dried over Na ₂ SO ₄ , filtered, concentrated and recrystallized from EtOAc/hexanes to give the desired product as a white solid (0.33 g, 78%) . ESI-MdesiredS m/z=440.07, 442.07 [MH] ^- .

Stage prom. 2h. To a solution of the compound of step 2g (1.8 g, 3.8 mmol) in DMSO (10 ml) at rt was added IBX (4.3 g, 15.3 mmol) and the mixture was stirred at rt for 20 h. Na ₂ S ₂ O ₃ NaHCO ₃ and a few drops of Et ₃ N and stirred at rt for 1 h. The mixture was extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound ( 1.65 g, 92%). ESI-MS m/z=470.04, 472.04 [Mn] ^- .

Intermediate 3

Stage prom. 3a. To a solution of intermediate 2 (1.76 g, 4.0 mmol) and trimethylsulfoxonium iodide (1.76 g, 8.0 mmol) in DMSO (20 ml) at 0 10 mmol). The resulting reaction mixture was stirred at rt for 1 h. The reaction mixture was quenched with aqueous NH ₄ Cl and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (1.36 g, 75%). ESI-MS m/z=452.07, 454.07 [MH] ^- .

Stage prom. 3b. In the mixed solution of the compound of the stage prom. 3a (78 mg, 0.17 mmol) in DMF (2.5 ml) was added NH ₄ Cl (17 mg, 0.32 mmol) and NaN ₃ (44 mg, 0.67 mmol) and then stirred at 60° C for 24 h. Dilute the mixture with EtOAc, wash with water and brine, dry over Na2SO4, filter, concentrate, chromatograph (silica, hexanes/EtOAc) to give the title compound as a white solid (73 mg, 88%) . ESI-MS m/z=495.08, 497.08 [MH] ^- .

Stage prom. 3 s. In the solution of the compound of the stage prom. 3b (0.20 g, 0.40 mmol) m-CPBA (0.27 g 77%, 1.2 mmol) was added to ΝΜΠ (2.0 ml) and stirred at rt in t/n. Add aqueous Na ₂ S ₂ O ₃ NaHCO ₃ and a few drops of Et3N and stir at rt for 1 h. The mixture is extracted with EtOAc, washed with water and brine, dried over Na2SO4, filtered, concentrated and purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.21 g, 98%). ESI-MS m/z=527.07, 529.07 [MH] ^- .

Stage prom. 3d. To a solution of Example 257 (540 mg, 1.02 mmol) in MeOH (2 ml) and THF (1 ml) was added Raney nickel (washed with MeOH, 50 mg). Connect a cylinder filled with hydrogen. The mixture was stirred for 2 h at rt. The mixture is filtered through a pad of celite and washed with MeOH. The filtrate was concentrated to give the title compound (440 mg, 86%). ESI-MS m/z=501.08, 503.08 [M-H] ^- .

Intermediate 4

Stage prom. 4a. In the solution of the compound of the stage prom. 2b (7.7 g, 77.18% in THF, 43 mmol) in anhydrous dichloromethane (20 mL) DBU (7.9 g, 5.2 mmol) and 1,1,2,2, 3,3,4,4,4-nonafluorobutane-1sulfonyl fluoride (14.4 g, 48 mmol). The reaction is maintained at 0° C. for 0.5 h before being concentrated to dryness. The residue is dissolved in hexane (70 ml). The solution was washed with HCl (0.5 M), water, NaHCO ₃ , brine and dried (Na ₂ SO ₄ ). It is filtered through a plug of silica gel, washed with hexane (300 ml) and concentrated to give a colorless oil (16.8 g, 92%). ¹ H-NMR (400 MHz, CDCl3) δ 5.13 (t, 1H), 4.85 (s, 2H), 2.67 (d, 2H), 2.46 (brs, 2H), 2.04 (dd, 2H), 1.72 (m, 2H), 1.58 (m, 2H).

Stage prom. 4b. In a mixture of compound stage prom. 4a (2.101 g, 5 mmol) and 4-methylmorpholine-4-oxide (0.703 g, 6.00 mmol) in acetone-water (4.5 ml/0.5 ml) osmium(VIII) oxide (0.628 mml, 2.5% in t-BuOH), and stirred at rt in t/n. Add Na ₂ S ₂ O ₃ (1.58 g, 10 mmol) and water (2 ml) and stir at rt for 30 min. The mixture is separated (EtOAc/water). The organic phase is washed with 1 N HCl, aqueous NaHCO ₃ , brine and dried (Na ₂ SO ₄ ). After filtration, the crude product is concentrated to obtain the desired product (2.24 g, 99%). ¹ H-NMR (400 MHz, CDCl ₃ ) δ 5.03 (t, 1H), 3.60 (s, 2H), 2.44 (brs, 2H), 1.99 (ddd, 4H), 1, 82 (m, 4H), 1.61 (m, 2H).

Stage prom. 4s. Suspension of compounds of the stage prom. 4b (1.84 g, 5.81 mmol), triphenylphosphine (0.063 g, 0.024 mmol) in THF (4 ml) degassed at rt followed by the addition of potassium tert-butoxide (1 M in THF, 5.32 ml, 5. 32 mmol). After 5 minutes, add the compound of step 1c (2.2 g, 4.84 mmol) in THF (9 ml) and stir at 60°C for 24 hours. After cooling, the mixture is diluted with MBTE (60 ml), filtered and washed with MBTE . The combined solution was washed with 0.5 N NaOH, brine and dried (Na ₂ SO ₄ ). It is filtered through a thin layer of silica (10 g silica gel) and washed with EtOAc (50 ml). The combined organic phase is concentrated in vacuo to give 2.5 g of crude product (110%).

Stage prom. 4d. In the solution of the compound of the stage prom. 4c (1.80 g, 3.81 mmol) m-CPBA (77 wt%, 2.14 g, 9.54 mmol) is added to ΝΜΠ (5 ml). It was stirred at rt for 20 h before addition of aqueous NaS2O3 (3 ml) followed by addition of aqueous NaHCO3 (3 ml) and MeOH (5 ml). The white solid is collected in vacuo and washed with aqueous NaHCO ₃ and water. This mixture was further recrystallized from MeOH to give the title compound (1.7 g, 87%). ESI-MS m/z=502.07, 504.07 [MH] ^- .

Intermediate 5

Complex SO ₃ pyridine (1.895 g, 11.9 mmol). The reaction mixture was stirred at rt for 3 h. It was diluted with EtOAc and washed with 1 M HCl and brine. The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by chromatography (silica, hexanes/acetone) to give the title compound (1.59 g, 3.18 mmol, 80% yield). ESI-MS m/z=500.05, 502.05 [MH] ^- .

Intermediate 6

The title compound was prepared using procedures similar to those described for Intermediate 1. ESI-MS m/z=297.99, 299.99 [MH] ^- .

Example 1

Stage 1a. To a solution of intermediate 3 (50 mg, 0.10 mmol) and (tert-butoxycarbonyl)-Lproline (26 mg, 0.12 mmol) in DMF (1.0 ml) was added DIPEA (0.051 ml, 0.30 mmol) and HATU (45 mg,

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0.12 mmol). It is stirred 2 h at rt and purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid (20 mg, 29%). ESI-MS m/z=698.19, 700.19 [MH] ^- .

Stage 1b. To a solution of the compound of step 1a (16 mg, 0.023 mmol) in THF (0.4 ml) was added HCl (4 M, 0.4 ml, 1.6 mmol). It is stirred 3 h at rt and concentrated to give the title compound as a white solid (14 mg, 99%). ESI-MS m/z=598.14, 600.14 [MH] ^- .

Stage 1c. To a solution of the compound of step 1b (12 mg, 0.019 mmol), formaldehyde (0.1 ml, 37% aqueous solution) and DIPEA (0.033 ml, 0.019 mmol), as well as a few drops of acetic acid in THF (0.5 ml ), add NaBH(OAc) ₃ (12 mg, 0.057 mmol). It is stirred 2 h at rt and saturated aqueous NaHCO ₃ is added. It is extracted with EtOAc, washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated, and the crude product is purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid (8 .2 mg, 71%). ESI-MS m/z=612.12, 614.12 [MH] ^- .

Example 3

TEA (4 drops) was added to a solution of the compounds of intermediate 4 (38 mg, 0.06 mmol) and sulfanyl chloride (8.5 mg, 0.072 mmol) in THF (0.5 ml) at rt, and the mixture was stirred at rt for 3 hours. It was concentrated and purified by preparative HPLC (C18 column, acetonitrile/water eluent) to give the title compound (13.5 mg, 38%). ESI-MS m/z=581.04, 583.04 [MH] ^- .

Example 4

Stage 4a. To a solution of intermediate 2 (880 mg, 2.0 mmol) in THF (10 ml) at 0°C was added prop-1-en-1-ylmagnesium chloride (0.5 M, 12 ml) and the solution was stirred at this temperature for 30 min before quenching it with NH ₄ Cl (20 ml). It is extracted with EtOAc, the organic phase is washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, ethyl acetate/hexanes) to give the Z isomer of the title compounds (252 mg, 26% yield) as a white solid and the E isomer (596 mg, 61%). ESI-MS m/z=502.09, 504.07 [MH] ^- .

Stage 4b. To a solution of the Z-isomer of step 4a (252 mg, 0.523 mmol) and NMO (123 mg, 1.05 mmol) in acetone (4 ml)/water (1 ml) was added OsO ₄ (4% in t-BuOH, 0.066 ml , 0.01 mmol), and the solution is stirred at rt in t/n. It is diluted with EtOAc, washed with Na ₂ S ₂ O ₃ , NaHCO ₃ , water and brine, and concentrated. This crude product was dissolved in THF (3 ml), m-CPBA (77 wt%, 234 mg, 1.1 mmol) was added and stirred at rt in t/n. After quenching with aqueous Na ₂ SO ₃ and aqueous NaHCO ₃ , it is extracted with EtOAc before drying and concentration. The crude product is crystallized from hot MeOH to give the title compound (223 mg, 78%, racemic) as a white solid. ESI-MS m/z=546.06, 548.06 [MH] ^- .

Example 5 _F he he

The title compound (racemic, white solid) was prepared from the E isomer of Step 4a following procedures similar to those described in Step 4b. ESI-MS m/z=546.06, 548.06 [MH] ^- .

Example 6

Stage 6a. In a mixture of compound stage prom. 2b (1.400 g, 4.67 mmol) and intermediate 6 (0.922 g, 5.14 mmol) in toluene (30 mL) triphenylphosphine (1.715 g, 6.54 mmol) was added at rt followed by DIAD (1.181 ml, 6.07 mmol). The mixture was stirred at 95° C. overnight, then cooled to rt and immediately purified by column chromatography (silica, hexanes/EtOAc) to give the title product as a white crystal (1.760 g, 90%). ESI-MS m/z=418.07, 420.06 [MH] ^- .

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Stage 6b. To a clear solution of the compound of step 6a (1.760 g, 4.19 mmol) in THF (40 mL) and water (0.5 mL) at rt was added NMO (2.455 g, 20.96 mmol) followed by dropwise addition of osmium tetroxide ( 4 wt% in water, 1.644 ml, 0.210 mmol).

The mixture was stirred at rt overnight. Additional osmium tetroxide (4 wt%, 1.644 ml, 0.210 mmol) is added. The yellow solution was stirred at rt overnight. Saturated Na ₂ S ₂ O ₃ is added to quench the reaction. After 20 min at rt, the mixture is diluted with THF. The aqueous layer is extracted from the THF extract (x 1). The combined organic layers are washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The solid residue is recrystallized from boiling MeOH (40 ml) to give the desired product as a white crystalline solid (1.620 g, 80%). ESI-MS m/z=484.04, 486.04 [MH] ^- .

Stage 6c. To a solution of the compound of step 6b (1.320 g, 2.72 mmol) and DIPEA (2.467 ml, 14.13 mmol) in DCM (8 ml) and DMSO (4.24 ml), cooled to 0°C, was added the sulfur trioxide complex and pyridine (1.340 g, 8.42 mmol). The resulting solution was stirred at 0° C. for 4 hours. The mixture was diluted with EtOAc/THF and then washed with aqueous 0.1 N HCl (x1), water (x 1) and brine (x 1). The organic layer is dried over Na ₂ SO ₄ (TV), filtered and concentrated. The solid was dissolved in DCM/THF (1/1) and purified by filtration through a short column (silica, hexanes/THF) to give the title product as an off-white foam (1.420 g, quant.). ESI-MS m/z=482.04, 484.04 [MH] ^- .

Stage 6d. Glyoxal (40% in water, 0.071 ml, 0.620 mmol) was added to a solution of the compound of step 6c (0.150 g, 0.310 mmol) in DMSO (3 ml) and 7 N ammonia in methanol (1.328 ml, 9.30 mmol) at rt. The resulting solution was stirred at RT overnight. Volatile substances are removed from the mixture. The remaining solution was immediately purified by HPLC (40-90% CH3CN in H2O) to afford the title compound as a white solid (42.0 mg, 26%). ESI-MS m/z=520.07, 522.07 [MH] ^- .

Example 7

Stage 7a. To a solution of the compound of step 6c (0.120 g, 0.248 mmol) in THF (4 mL) cooled to -78°C, methylmagnesium bromide (3 M in Et2O, 0.413 mL, 1.240 mmol) was added dropwise. The reaction mixture was stirred at -78° C. for 30 minutes. Additional methylmagnesium bromide (3 M in Et ₂ O, 0.413 mL, 1.240 mmol) is added. The reaction mixture was stirred at -78°C for 2 h, after which it was allowed to warm to rt and quenched with saturated NH4Cl solution. The mixture is diluted with THF and water. The organic layer is washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was dissolved in DMSO (4 ml) and purified by HPLC (40-90% ACN in water) to give the title compound as a white solid (39.0 mg, 31%, racemic). ESI-MS m/z=498.07, 500.07 [MH] ^- .

Example 9

F

Stage 9a. A solution of ethyl 2-(diethoxyphosphoryl)acetate (673 mg, 3.0 mmol) in THF (10 ml) was treated with NaH (60 w/w%, 120 mg, 3.0 mmol) at rt for 30 min before adding intermediate 5 (502 mg, 1.0 mmol). It is stirred in t / n at rt. It is diluted with EtOAc and washed with aqueous NH ₄ Cl and brine. The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by chromatography (silica, hexanes/acetone) to give the title compound (280 mg, 49%). ESI-MS m/z=570.09, 572.09 [MH] ^- .

Stage 9b. NaBH ₄ (16 mg, 0.42 mmol) was added to a solution of step 9a (80 mg, 0.14 mmol) in THF (1 ml)/EtOH (1 ml) and stirred at rt for 2 h, after which a second portion of NaBH ₄ (20 mg). It is stirred for an additional 3 hours, after which it is quenched with water and extracted with EtOAc. The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by preparative HPLC (C18 column, acetonitrile/water) to give the title compound (13.5 mg, 38%). ESI-MS m/z=530.08, 532.08 [MH] ^- .

Example 10

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Stage 10a. To a solution of triethylphosphonoacetate (0.520 ml, 2.60 mmol) in THF (5.0 ml) at 0°C was added NaH (0.104 g, 60%, 2.6 mmol). The resulting reaction mixture was stirred at 0° C. for 30 minutes. A solution of intermediate 2 (0.15 g, 0.34 mmol) in THF (2.0 ml) was added and stirred at rt for 2 h. The reaction mixture was quenched with aqueous NH ₄ Cl and the mixture was extracted with EtOAc, washed with water and brine . The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.50 g, 98%). ESI-MS m/z=508.10, 510.10 [MH] ^- .

Step 10b. To a solution of the compounds of step 10a (0.32 g, 0.627 mmol) in THF (5.0 ml) at -78°C was added DibAL-H (2.5 ml, 1.0 M solution in hexanes, 2.5 mmol) . The resulting reaction mixture was stirred at -78° C. for 1 hour. The reaction mixture was treated with aqueous potassium sodium tartate for 3 hours and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (132 mg, 45%). ESI-MS m/z=466.06, 468.07 [MH] ^- .

Stage 10s. To a mixture of the compound of step (43 mg, 0.092 mmol) and NMO (64 mg, 0.55 mmol) in acetone (3.0 ml) at rt was added osmium tetroxide (0.58 ml, 4% in water, 0.092 mmol), and the mixture was stirred at rt for 16 h. It was quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO ₃ and brine, dried with Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC (column C18, acetonitrile/water) to give the title compound (18 mg, 37%, racemic). ESI-MS m/z=578.07, 580.07 [M+HCO2] ^- .

Example 11

Stage 11a. To a solution of intermediate 2 (880 mg, 2.0 mmol) in THF (10 ml) was added vinylmagnesium chloride (1.6 M in THF, 3.75 ml, 6.0 mmol). It is stirred for 30 min before aqueous NH4Cl is added. It is extracted twice with EtOAc. The organic phase is washed with brine and dried (Na ₂ SO ₄ ). After concentration to 10 ml, the mixture is filtered under vacuum to obtain the desired product (616 mg, 66%). ESI-MS m/z=466.08, 468.08 [MH] ^- .

Stage 11b. To a solution of step 11a (94 mg, 0.2 mmol) in THF (2 ml) was added m-CPBA (77 wt.%, 224 mg, 1.0 mmol) and stirred at rt in t/n. After quenching with aqueous Na ₂ SO ₃ and aqueous NaHCO ₃ , it is extracted with EtOAc, dried and concentrated. The crude product was purified by chromatography (silica, ethyl acetate/hexanes) to give the title compounds (75 mg, 73% yield) as a white solid. ESI-MS m/z=514.08, 516.08 [MH] ^- .

Stage 11c. To a solution of step 11b (75 mg, 0.14 mmol) in DMF (1 mL) was added NaN3 (29 mg, 0.44 mmol), NH ₄ Cl (8 mg, 1.5 mmol) and stirred at 55°C for t/n. After cooling, it is diluted with EtOAc and filtered. After concentration, it was purified by preparative HPLC (C18 column, acetonitrile/water eluent) to give the title compound (13.5 mg, 38%, racemic) as a white solid. ESI-MS m/z=557.04, 559.04 [MH] ^- .

Example 14

To a solution of intermediate 3 (50 mg, 0.10 mmol) and (S)-tetrahydrofuran-2-carboxylic acid (14.43 μl, 0.149 mmol) in DMF (1 ml) was added EDA (38 mg, 0.20 mmol ) and DMAP (36 mg, 0.12 mmol). The reaction mixture was stirred at rt for 2 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (14 mg, 23%, single enantiomer). ESI-MS m/z=599.125, 601.122 [MH] ^- .

Example 18

To a solution of intermediate 3 (75 mg, 0.15 mmol), DIPEA (52.1 μl, 0.298 mmol) and cyclohex-3-ene-1-carboxylic acid (18.8 mg, 0.15 mmol) in DMF ( 1.0 ml) at rt add HATU (68.0 ml, 0.18 mmol) and stir at rt for 4 days. It is purified by preparative HPLC (C-33 041670

18, acetonitrile/water) to give the title compound as a white solid (65 mg, 71%). ESI-MS m/z=609.14, 611.14 [MH] ^- .

Example 19

A mixture of the compound from Example 18 (40 mg, 0.065 mmol) and NMO (23 mg, 0.196 mmol) in acetone/water (2.0/2.0 ml) at rt was treated with osmium tetroxide (0.042 ml, 4% in water , 0.0065 mmol) at rt for 16 h. It is quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC (column C18, acetonitrile/water) to give the title compound (26 mg, 61%, single isomer, stereochemistry not determined). ESI-MS m/z=643.15, 645.15 [MH] ^- .

Example 20

Stage 20a. To a suspension of (methoxymethyl)triphenylphosphonium chloride (3.43 g, 10 mmol) in THF (16 ml) at 0°C was added t-BuOK (1.68 g, 15 mmol). The resulting mixture was stirred at rt for 30 min. A solution of intermediate 2 (2.2 g, 5.0 mmol) in THF (4.0 ml) was added and stirred at rt for 20 h. It was quenched with aqueous NH4Cl and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (2.08 g, 89%). ESI-MS m/z=466.13, 468.13 [MH] ^- .

Stage 20b. To a solution of the compound of step 20a (1.1 g, 2.35 mmol) in THF (10 ml) at rt was added concentrated HCl (1.5 ml) and stirred at rt for 2 h. It was concentrated in vacuo to remove most of the THF , and the residue is extracted with EtOAc. The organic phase is washed with water, 10% K2CO3 and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (0.95 g, 89%). ESI-MS m/z=452.07, 454.07 [MH] ^- .

Stage 20s. To a solution of the compound of step 20b (0.27 g, 0.59 mmol) in THF (6.0 ml) at 0°C was added vinylmagnesium bromide (2.37 ml, 1 M in THF, 2.37 mmol). The resulting reaction mixture was stirred at 0° C. for 1 hour. The reaction mixture was quenched with aqueous NH4C1 and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a mixture of diastereomers (50 mg, 17%). ESI-MS m/z=480.08, 482.08 [MH] ^- .

Stage 20d. To a mixture of the compound of step 20c (50 mg, 0.104 mmol) and NMO (73 mg, 0.62 mmol) in acetone (3.0 ml) was added osmium tetroxide (0.66 ml, 4% in water, 0.104 mmol) at rt. , and the mixture was stirred at rt for 16 h. It was quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC using C18 column and acetonitrile/water as eluent to give the title compound as a mixture of diastereomers (21 mg, 37%). ESI-MS m/z=546.06, 548.06 [MH] ^- .

Example 21

Stage 21a. BuLi (2.6 M in hexanes, 1.091 ml, 2.73 mmol ). The resulting clear solution was stirred at -78°C for 1 hour. A solution of intermediate 2 (0.150 g, 0.341 mmol) in THF (1 ml) was added at -78°C. The mixture was stirred at -78°C for 1 h, after which it was allowed to warm to rt and stirred at rt for 30 min. Saturated NH4C1 solution is added to quench the reaction. The mixture is diluted with EtOAc and water. The organic layer is washed with brine (x2), dried over Na2SO4 (tv), filtered and concentrated. The residue was purified by column chromatography (silica, hexanes/EtOAc) to give the desired product as a yellow solid (0.152 g, 85%). ESI-MS m/z=522.11, 524.11 [MH] ^- .

Stage 21b. To a solution of the compound of step 21a (140 mg, 0.267 mmol) in ethyl acetate (12 ml) at rt was added Lindlar's catalyst (114 mg, 0.053 mmol). The suspension is stirred with the H ₂ balloon at RT overnight. LC-MS shows ~20% conversion. The mixture is filtered through a thin layer of celite.

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The filtrate is concentrated. The residue was purified by column chromatography (silica, hexanes/acetones) to give the desired product as a white solid (23.5 mg, 17%).

ESI-MS m/z=524.11, 526.11 [MH] ^- .

Stage 21c. To a clear solution of the compound of step 21b (23.5 mg, 0.045 mmol) in THF (2.80 mL) and water (0.200 mL) at rt was added NMO (26.2 mg, 0.223 mmol) followed by osmium tetroxide (4% in water, 0.057 ml, 8.94 µmol). The solution was stirred at rt over the weekend, followed by stirring at 55° C. for 2 nights. Additional osmium tetroxide (4% in water, 0.057 ml, 8.94 µmol) is added. The mixture was stirred at 55° C. overnight, after which it was quenched with saturated Na ₂ S ₂ O ₃ solution and diluted with THF. The organic layer is washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was dissolved in DMSO (2 ml) and purified by HPLC (40-90% ACN in water) to give the title compound as a white solid (3.5 mg, 14%). ESI-MS m/z=556.12, 558.11 [MH] ^- .

Example 22

The title compound (44 mg, 84%) was prepared following the procedure described in Example 14 from Intermediate 3 and cyclopent-3-ene-1-carboxylic acid. ESI-MS m/z=641.13, 643.13 (M+HCO2)·.

Example 26

Stage 26a. The target compound is obtained from the compound of example 11, following a procedure similar to that described in step prom. 3d, ESI-MS m/z=531.09, 533.09 [MH] ^- .

Step 26b. A solution of the compound of step 26a (50 mg, 0.091 mmol) and DMAP (60 mg, 0.49 mmol) in THF-water (1.0/0.1 ml) at rt was treated with MsCl (38 mg, 0.33 mmol) at rt for 1 h. It is concentrated in vacuo and purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (13 mg, 23%). ESI-MS m/z=609.07, 611.07 [MH] ^- .

Example 27

Stage 27a. The title compound is prepared following a procedure similar to that described in Example 14 from Intermediate 3 and (tert-butoxycarbonyl)-L-asparagine. ESI-MS m/z=715.18, 717.18 [MH] ^- .

Stage 27b. The compound of step 27a is treated with HCl (4 M in dioxane) for 2 h at rt. This is concentrated to give the title compound as a white solid HCl salt. ESI-MS m/z=615.12, 617.12 [M-H-56] ^- .

Example 28

Stage 28a. Adonitol (1.0 g, 6.57 mmol) and pyridine hydrochloride (1.215 g, 10.52 mmol) are gently mixed and heated to 150°C for 4 h. The crude product is purified by chromatography (silica, EtOAc/MeOH) to obtain the target compound as a colorless rubbery substance (882 mg, 100%).

Step 28b. To a solution of the compound of step 28a and 2,2-dimethoxypropane (3.23 mL, 26.3 mmol) in acetone (26.303 mL) was added PTCA (250 mg, 1.315 mmol) and then stirred at rt for 30 min. The reaction mixture was quenched with aqueous NaHCO ₃ and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, dichloromethane/MeOH) to give the title compound as a colorless oil (852 mg, 74%).

Stage 28c. To a solution of the compound of step 28b (327 mg, 1.88 mmol) in acetonitrile (2 ml) and water (2 ml) was added TEMPO (59 mg, 0.375 mmol) and iodobenzene diacetate (1.21 g, 3.75 mmol). The reaction mixture was stirred at rt for 1 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude acid was used without further purification (454 mg, 50% purity, 64%).

Stage 28d. To a solution of intermediate 3 (150 mg, 0.30 mmol) and the compound of step 28c (168 mg, 0.447 mmol, 50% purity) in DMF (3 mL) was added EDA (114 mg, 0.60 mmol) and DMAP (109 mg, 0.12 mmol). The reaction mixture was stirred at rt for 2 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (117 mg, 58%). ESI-MS m/z=717.153, 719.151 [M+CO2H] ^- .

Example 29

To a solution of the compound of example 28 (115 mg, 0.171 mmol) in THF (5 ml) and methanol (10 ml) was added HCl (4 ml, 2 M aq., 8 mmol). The reaction mixture is heated to 60°C for 1 h, the Reaction mixture is extracted with EtOAc, washed with NaHCO ₃ , water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (76 mg, 70%). ESI-MS m/z=677.119, 679.117 [M+CO2H] ^- .

Example 32

Stage 32a. BF ₃ diethyl etherate ( 1.11 ml, 8.73 mmol). Stir for 1 h at -78°C, then warm to rt for 1 h. The reaction mixture is quenched with NaHCO ₃ (aqueous). The crude product is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/EtOAc) to give the title compound as a white solid (2.976 g, 80%). ESI-MS m/z=511.8, 513.8 [MH] ^- .

Step 32b. To a solution of the material from step 32a (2.976 g, 5.79 mmol) in ΝΜΠ (29 ml) was added m-CPBA (3.89 g, 17.37 mmol, 77%). The reaction mixture was stirred at rt for 18 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The residue is triturated with methanol and filtered to give the title compound as a white solid (2.50 g, 4.58 mmol). ESI-MS m/z=544.076, 546.074 [MH] ^- .

Example 33

To a solution of the compound of Example 32 (2.50 g, 4.58 mmol) in THF (27 mL) and methanol (10 mL) was added LiOH (9.16 mL, 18.32 mmol, 2 M aq.). It is stirred at rt for 6 h. The reaction mixture is acidified to pH 3 with HCl (2 M, aq.). The crude product is extracted with EtOAc and washed with brine. The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound (2.40 g, 99%). ESI-MS m/z=498.068, 500.066 [M-H] ^- .

Example 34

Step 34a. t-BuOK (1.12 d, 10 mmol). The resulting reaction mixture was stirred at rt for 1 h. The reaction mixture was quenched with aqueous NH4Cl and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (1.36 g, 75%). ESI-MS m/z=452.07, 454.07 [MH] ^- .

Step 34b. To a mixture of the compound of step 34a (0.12 g, 0.264 mmol) and cyclopent-3-en-1-ol (0.523 ml, 6.61 mmol) was added potassium tert-butoxide (297 mg, 2.64 mmol) at rt and heated to 80°C for 1 hour,

- 36 041670 then maintained at 50°C for 16 hours. The reaction mixture was quenched with aqueous NH4C1 and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound (82 mg, 57%). ESI-MS m/z=536.11, 538.11 [MH] ^- .

Stage 34c. Osmium tetroxide (0.29 ml, 4% in water, 0.046 mmol) was added to a mixture of the compound of step 34b (50 mg, 0.093 mmol) and NMO (65.3 mg, 0.558 mmol) in acetone (2.0 ml) at rt. , and the mixture was stirred at rt for 2 days. It is quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO 3 and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC using a C18 column and acetonitrile/water as eluent to give the title in the title compound, the structure of which is determined experimentally (11 mg, 19%). ESI-MS m/z=602.12, 604.12 [MH] ^- .

Example 35

The title compound (12 mg, 21%) was isolated from Example 34. ESI-MS m/z=602.12, 604.12 [MH] ^- .

Example 36

Step 36a. Croton alcohol (341 µl, 4.00 mmol) was added to a suspension of NaH (160 mg, 60%, 4.00 mmol) in DMF (5.0 ml) at 0°C and stirred at rt for 30 min. Step 34a (182 mg, 0.40 mmol) was added and stirred at rt for 16 h. Quenched with aqueous NH4Cl, extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by chromatography (dioxide silicon, hexanes/EtOAc) to obtain the target product (195 mg, 93%). ESI-MS m/z 524.11, 526.11 [MH] ^- .

Step 36b. Osmium tetroxide (1.15 ml, 4% in water, 0.181 mmol) and the mixture was stirred at rt for 2 days. It is quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO ₃ and brine, dried over Na 2 SO 4 , filtered, concentrated and purified by preparative HPLC using a C18 column and acetonitrile/water as eluent to give the title compounds, the structure of which is determined experimentally (36 mg, 17%). ESI-MS m/z=590.11, 592.11 [MH] ^- .

Example 37

Stage 37a. In the solution of the compound of the stage prom. 2g (1.9 g, 4.30 mmol) in ΝΜΠ (12 ml) m-CPBA (2.89 g, 12.90 mmol) was added at rt. The mixture was stirred at rt overnight. It is quenched with an aqueous solution of Na ₂ S ₂ O ₃ and an aqueous solution of NaHCO ₃ with a few drops of triethylamine. The mixture is extracted with EtOAc. The organic layer is washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The solid residue is recrystallized from MeOH to give the desired product as a white solid (1.8 g, 88%). ESI-MS m/z=472.06, 474.06 [M-H] ^- .

Stage 37b. To a solution of the compound of step 37a (1.8 g, 3.8 mmol) in DMSO (10 mL) was added IBX (4.3 g, 15.3 mmol) at rt. The mixture was stirred at 45° C. for 20 hours. An aqueous solution of Na ₂ S ₂ O ₃ and an aqueous solution of NaHCO ₃ were added with a few drops of Et ₃ N. The mixture was stirred at rt for 1 hour. It was extracted with EtOAc. The organic layer was washed with water and brine, dried over Na2SO4, filtered and concentrated to give the title compound (1.65 g, 92%). ESI-MS m/z=470.04, 472.04 [MH] ^- .

Stage 37c. To a solution of the compound of step 37b (104 mg, 0.220 mmol) in THF (2 ml) at 5-10° C., methyl magnesium bromide (3 M in ether, 367 μl, 1.102 mmol) is added dropwise. Additional THF (2.5 ml) was added and the mixture was stirred at rt for 1 h. It was diluted with EtOAc, washed with water, aqueous Na ₂ SO ₃ and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by preparative TLC (silica, hexanes/EtOAc) to give the title compound (85 mg, 79%). ESI-MS m/z=486.08, 488.08 [MH] ^- .

Stage 37d. To a clear solution of the compound of step 37c (30.0 mg, 0.061 mmol) in THF/MeOH (1/1, 2.0 mL) at rt was added 10% Pd/C (6.5 mg, 6.15 µmol) in one portion . The suspension was purged 3 times with H ₂ and then stirred with the H ₂ balloon at RT overnight. The suspension is then stirred at rt

- 37 041670 nitrogen atmosphere (60 psi) for 4 hours Add additional 10% Pd/C (13.0 mg, 12.3 µmol). The slurry was purged with H ₂ 3 times and then stirred at rt under H ₂ (~15 psi) over the weekend. The mixture is filtered through a thin layer of celite. Volatile substances are removed from the filtrate. The solid residue was triturated with DCM to give the title compound as a white solid (20.0 mg, 72%). ESI-MS m/z=452.11.453.11 [MH] ^- .

Example 38

Step 38a. The solution of the compound stage prom. 3a (214 mg, 0.47 mmol) in THF/water (3.0/1.0 mL) was treated with TFA (0.40 mL) at rt for 6 h. It was concentrated in vacuo to remove most of the THF. The residue is extracted with EtOAc. The organic phase is washed with water, 10% K ₂ CO ₃ and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (0.20 g, 90%). ESIMS m/z=470.08, 472.08 [MH] ^- .

Step 38b. To a solution of the compound of step 38a (0.340 g, 0.720 mmol) in DMSO (6 mL) was added IBX (0.303 g, 1.081 mmol) at rt. The resulting milky mixture was stirred at rt for 2 h. The mixture was diluted with EtOAc and water. The organic layer is washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The solid residue was dried in vacuo to give the desired product as a white solid (0.324 g, 96%). ESI-MS m/z=468.05, 470.05 [MH] ^- .

Stage 38c. To a suspension of the compound of step 38b (0.324 g, 0.690 mmol) in t-BuOH (6.0 mL) and water (2.0 mL) at rt was added potassium phosphate monobasic (0.657 g, 4.83 mmol) followed by 2methyl- 2-butene (1.826 ml, 17.24 mmol). Sodium chlorite (80%, 0.702 g, 6.21 mmol) is added to the suspension in one portion. The resulting clear solution was stirred at rt for 1 h. It was diluted with MTBE and 1.0 M NaOH (8 ml). The mixture was diluted with EtOAc, washed with aqueous 0.5 N HCl (x1) and then with brine (x1). The organic phases are dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was dried in vacuo to give the desired product as a white solid (0.306 g, 91%). ESI-MS m/z=484.05, 486.05 [MH] ^- .

Stage 38d. To a suspension of the compound of step 38c (50.0 mg, 0.103 mmol), but-2-en-1-amine hydrochloride (12.18 mg, 0.113 mmol) and DIPEA (0.054 ml, 0.309 mmol) in acetonitrile (3 ml) at HATU (47.0 mg, 0.123 mmol) was added in one portion. The resulting pale milky solution was stirred at RT overnight. Additional DIPEA (0.054 ml, 0.309 mmol) and HATU (47.0 mg, 0.123 mmol) are added. The resulting solution was stirred at rt for 1 h and then at 55°C for 2 h. Volatiles were removed from the mixture. The residue was purified by column chromatography (silica, hexanes/EtOAc) to give the desired product as a white solid (7.0 mg, 12%). ESI-MS m/z=583.11, 585.11 [MH] ^- .

Stage 38e. To a clear solution of the compound of step 38d (7.0 mg, 0.013 mmol) in THF (2.0 mL) and water (0.14 mL) at rt was added NMO (7.61 mg, 0.065 mmol) followed by osmium tetroxide ( 0.083 ml, 0.013 mmol). The solution is stirred at 50° C. overnight. The mixture is quenched with saturated Na ₂ S ₂ O ₃ solution and diluted with THF. The organic layer is washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was dissolved in DMSO (1 ml) and purified by HPLC (40-90% ACN in water) to give the title compound as a white solid (3.0 mg, 38%, racemic mixture of diastereomers). ESI-MS m/z=649.13, 651.13 [M+HCO2] ^- .

Example 42

Step 42a. To a solution of the compound of step 82a (173 mg, 0.37 mmol) in CH ₂ Cl ₂ (3.7 ml) at rt was added (2)-but-2-ene-1,4-diylacetate (234 μl, 1.479 mmol) and a second generation Grubbs-Hoveid catalyst (23.1 mg, 0.037 mmol), then the mixture is degassed and refluxed for 24 h. The mixture is concentrated and the residue is purified by chromatography (silica, hexanes/EtOAc) to give the title compound in as a white solid (160 mg, 80%). ESI-MS m/z=584.10, 586.10 (M+HCO2) ^- .

Step 42b. Potassium carbonate (10.5 mg, 0.076 mmol) was added to a solution of the compound of step 42a (20.5 mg, 0.038 mmol) in MeOH (1.3 ml) at rt, then the mixture was kept at rt for 1 h. The reaction mixture was diluted water and extracted with CH ₂ Cl ₂ , the organic phases are dried over Na ₂ SO ₄ and concentrated to obtain the target product (19 mg) without further purification. ESI-MS m/z=542.10, 544.10 (M+HCO2) ^- .

- 38 041670

Stage 42c. To a solution of the compound of step 42b (18.9 mg, 0.038 mmol) in acetone/H ₂ O (0.8 ml, 4:1) was added NMO (26.7 mg, 0.228 mmol) and osmium tetroxide (298 μl, 0.083 mmol ). The mixture is kept at 50°C for 2 days. It was quenched with aqueous Na2SO3, extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC using a C18 column and acetonitrile/water as eluent to give the title compound (3.9 mg, 18%). ESI-MS m/z=608.10, 610.10 (M+HCO2)·.

Example 51

Step 51a. BuLi (2 .6 M in hexanes, 1.819 ml, 4.55 mmol). The resulting clear solution was stirred at -78°C for 0.5 h. A solution of intermediate 2 (500 mg, 1.137 mmol) in THF (3 ml) was added dropwise at -78°C. The mixture is stirred at 78°C for 1 h, then quenched with a saturated solution of NH ₄ Cl. It was allowed to warm to rt and diluted with EtOAc and water. The organic layer is washed with brine (x1), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was purified by column chromatography (silica, hexanes/EtOAc) to give the desired product as a white solid (0.440 g, 63%). ESI-MS m/z=614.14, 616.14 [MH] ^- .

Step 51b. To a solution of the compound of step 51a (0.140 g, 0.227 mmol) in ethyl acetate (10 mL) was added Lindlar's catalyst (0.097 g, 0.045 mmol) at rt. The suspension is stirred with the H ₂ balloon at RT for 2 hours. The mixture is filtered through a thin pad of celite. The filtrate is concentrated. The residue was purified by column chromatography (silica, hexanes/EtOAc) to give the desired product as a white solid (96.0 mg, 68%). ESI-MS m/z=616.15.618.15 [MH] ^- .

Step 51c. To a clear solution of the compound of step 51c (46.0 mg, 0.074 mmol) in DCM (2 mL) was buffered to pH 7 (0.4 mL) at rt followed by DDC (33.8 mg, 0.149 mmol). The biphasic mixture was stirred at rt for 3 h. It was quenched with saturated NaHCO ₃ solution and diluted with DCM. The organic layer is dried over Na2SO4 (tv), filtered and concentrated. The residue was purified by chromatography (silica, hexanes/EtOAc) to give the desired product as a white solid (20.2 mg, 54%). ESI-MS m/z=496.09, 498.09 [MH] ^- .

Stage 51d. To a clear solution of the compound of step 51c (45.6 mg, 0.092 mmol) in THF (5.60 ml) and water (0.56 ml) at rt was added NMO (53.6 mg, 0.458 mmol) followed by the addition of osmium tetroxide in t-butanol (2.5%, 0.186 ml, 0.018 mmol). The solution was stirred at rt for 4 h and then at 50° C. for 2 nights. The mixture was quenched with saturated Na2S2O3 and diluted with THF. The organic layer is washed with brine (x2), dried over Na ₂ SO ₄ (tv), filtered and concentrated. The residue was dissolved in DMSO (2 ml) and purified by HPLC (30-90% ACN in water) to give the title compound as a white solid (15.0 mg, 29%). ESI-MS m/z=608.09, 610.08 [M+HCO2] ^- .

Example 52

To a solution of the compound of Example 33 (50 mg, 0.094 mmol), 3-methylbut-2-en-1-amide hydrochloride (12.6 mg, 0.103 mmol) and DIPEA (0.5 mg, 0.282 mmol) in DMF (0. 9 ml) add a solution of HATU (54 mg, 0.141 mmol) in DMF (0.5 ml). Stirred for 2 h at rt. The crude reaction mixture was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (40 mg, 71%). ESI-MS m/z=596.8, 598.8 [MH] ^- .

Example 56

To a solution of Example 52 (40 mg, 0.067 mmol) in acetone (5 mL) was added OsO ₄ (0.085 mL, 4 w/w% in water, 0.013 mmol) and NMO (19.6 mg, 0.167 mmol). It is stirred for 48 hours at rt, then evaporated on silica. The residue was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (18 mg, 43%). ESIMS m/z=630.8, 632.8 [MH] ^- .

- 39 041670

Example 61

Step 61a. To a mixture of the compound of Example 34 (0.030 g, 0.050 mmol) and sodium bicarbonate (8.4 mg, 0.10 mmol) in THF-water (1.5/0.5 ml) was added sodium periodate (0.032 g, 0.150 mmol) and stirred at rt for 16 h. It is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and used in the next step without further purification. ESI-MS m/z=600.10, 602.10 [MH] ^- .

Step 61b. To a solution of the compound of step 61a (30 mg, 0.050 mmol) in THF-MeOH (1.5/0.5 ml) at 0°C was added NaBH ₄ (7.57 mg, 0.20 mmol), then stirred at 0° C for 30 min. It is quenched with aqueous NH ₄ Cl, extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC using a C18 column and acetonitrile/water as eluent to give the title compound (7.6 mg, 25%). ESI-MS m/z=604.13, 606.13 [MH] ^- .

Example 64

Step 64a. To a solution of intermediate 5 (250 mg, 0.50 mmol) in THF (5 ml) in an ice bath was added vinylmagnesium chloride in THF (1.6 M, 1.25 ml, 2.0 mmol) and stirred at rt at for 1 h, after which it is quenched with aqueous NH ₄ Cl. It is extracted with EtOAc and washed with brine. After drying (Na ₂ SO ₄ ), it is concentrated to give the desired crude compound, which is used in the next step without further purification. ESI-MS m/z=528.07, 530.07 [MH] ^- .

Step 64b. To a solution of the crude compound of step 64a (not more than 0.3 mmol) in acetone (5 ml) and water (1 ml) was added NMO (67 mg, 0.57 mmol) and OsO ₄ (4% in water, 0.12 ml , 0.02 mmol) and stirred at rt for 2 days, after which it was quenched with aqueous Na ₂ S ₂ O ₃ ; it is extracted with EtOAc, then dried and concentrated. The crude product was purified by chromatography (silica, acetone/hexanes) to give the title compounds as a mixture of two racemic pairs (122 mg, 57%, white solid). ESI-MS m/z=562.08, 564.08 [MH] ^- .

Example 70

Step 70a. HATU (135 mg, 0.355 mmol). The reaction mixture was stirred at rt for 3 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the desired product as a white solid (54 mg, 34%).

Step 70b. To a solution of the compound of step 70a (54 mg, 0.08 mmol) in THF (2 mL) was added HCl (2 mL, 8 mmol, 4 M in dioxane). The reaction mixture was stirred for 2 hours, then evaporated to give the crude product, which was used without further purification.

Stage 70s. To a solution of the compound of step 70b (23 mg, 0.04 mmol) and DIPEA (0.017 ml, 0.1 mmol) in DMF is added isopropyl chloroformate (0.048 ml, 0.048 mmol, 1 M in toluene). Stir for 15 min at rt. The crude reaction mixture was purified by chromatography (preparative HPLC, acetonitrile/water) to give the title compound as a white solid (3.8 mg, 14%). ESI-MS m/z=660.0, 662.0 [M+H]+.

Example 74

To a solution of the compound of example 8 (62 mg, 0.092 mmol) in DMSO (0.6 ml) at rt was added IBX (38.6 mg, 0.138 mmol) and stirred at 45°C for 16 h. It was quenched with MeOH and purified by preparative HPLC using C18 column and acetonitrile/water as eluent to give the title compound (11 mg, 18%). ESI-MS m/z=670.13, 672.13 [MH] ^- .

- 40 041670

Example 76

Step 76 To a solution of intermediate 3 (95.0 mg, 0.189 mmol) and (2S,3R)-1-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid (43.7 mg, 0.189 mmol) in DMF ( 3 ml) DIPEA (0.099 ml, 0.567 mmol) was added at rt followed by HATU (108 mg, 0.283 mmol). The resulting clear solution was stirred at RT for 1 h. Volatiles were removed from the mixture.

The residue was taken up in DCM with some THF and purified by column chromatography (silica, DCM/MeOH) to give the title compound as a colorless sticky oil (128 mg, 95%). ESI-MS m/z=760.20, 762.20 [M+HCO2]-.

Example 78

Step 78a. To a solution of Example 76 (0.064 g, 0.090 mmol) in THF (2 mL) was added HCl (4 M in 1,4-dioxane, 0.900 mL, 3.60 mmol) at rt. The resulting clear solution was stirred at rt for 3 h. It was made into a suspension. The mixture is concentrated. The solid residue is immediately used in the next step. ESI-MS m/z=660.15, 662.15 [M+HCO2]-.

Step 78b. To a solution of the compound of step 78a (0.090 g) in DMF (2.0 ml) at rt was added DIPEA (0.157 ml, 0.900 mmol) followed by a solution of methyl chloroformate (6.97 μl, 0.090 mmol) in DMF (0.1 ml) . The resulting yellow solution was stirred at RT for 1 h, after which volatiles were removed from it. The residue was dissolved in DMSO (2 ml) and purified by HPLC (40-90% ACN in water) to give the title compound as a white solid (20.0 mg, 33% in 2 steps). ESI-MS m/z=718.16, 720.16 [M+HCO2]-.

Example 82

Step 82a. To a solution of the compound of step 150d (210 mg, 0.50 mmol) in THF (5 ml) in an ice bath was added vinylmagnesium chloride in THF (1.6 M, 0.94 ml, 1.5 mmol). It is stirred at rt for 1 h, after which it is quenched with aqueous NH4Cl. It is extracted with EtOAc and washed with brine. After drying (Na ₂ SO ₄ ), it was concentrated and the residue was purified by chromatography (silica, EtOAc/hexanes) to give the title compound (201 mg, 94%) as a white solid. ESI-MS m/z=448.08, 450.08 [MH]-.

Step 82b. To a solution of the compound of step 82a (152 mg, 0.34 mmol) in acetone (1 ml) and water (0.2 ml) was added NMO (79 mg, 0.68 mmol) and OsO ₄ (2.5% in t- BuOH, 0.12 ml, 0.007 mmol). It is stirred at rt in t/n, after which it is quenched with aqueous Na ₂ S ₂ O ₃ . It is extracted with EtOAc, dried and concentrated. The crude product is crystallized from MeOH to give the title compound (141 mg, 84%, racemic) as a white solid. ESI-MS m/z=514.08.516.08 [MH]-.

Example 106

The title compound (single enantiomer) was isolated upon preparation of the compound of Example 13. ESI-MS m/z=645.11, 647.11 [M-H]-.

Example 108

The title compound (single enantiomer, structure determined experimentally) was isolated from the compound of Example 4 by SFC chromatography.

- 41 041670

The title compound (single enantiomer, structure determined experimentally) was isolated from the compound of Example 5 by SFC chromatography.

Example 120

Example 109

To a solution of the compound of example 119 (50 mg, 0.083 mmol) in THF (1 ml) and water (0.67 ml) at rt was added LiOH (0.33 ml, 0.67 mmol, 2 M aq). The reaction mixture is stirred for 1 h, then acidified to pH 3 with 2 M HCl. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound as a white solid (40 mg, 82%). ESI-MS m/z=586.8, 588.8 [MH] ^- .

Example 121

Step 121a. Potassium 2-methylpropan-2-olate (630 mg, 5.62 mmol), then the mixture is stirred at 60° C. for 20 h. It is cooled to rt, quenched with aqueous NH ₄ Cl, extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by chromatography (silica, hexanes/EtOAc) to give the desired product (98 mg, 45%). ESI-MS m/z=572.15, 574.15 [MH] ^- .

Step 121b. To a solution of the compound of step 121a (95 mg, 0.165 mmol) in ΝΜΠ (1.5 ml) was added mCPBA (0.167 g, 77%, 0.745 mmol) and stirred at rt for 20 h. Aqueous Na ₂ S ₂ was added O ₃ NaHCO ₃ and a few drops of Et ₃ N and stirred at rt for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na2SO4, filtered, concentrated and purified by preparative HPLC using a C18 column and acetonitrile/water as eluent to obtain the title compound (44 mg, 44%). ESI-MS m/z=604.14, 606.14 [M-H] ^- .

Example 124

Step 124a. To a suspension of methyltriphenylphosphonium bromide (0.24 g, 0.67 mmol) in THF (1.0 ml) at 0°C was added t-BuOK (0.11 g, 1.0 mmol). The resulting reaction mixture was stirred at rt for 30 min. A solution of intermediate 2 (0.15 g, 0.34 mmol) in THF (1.0 ml) was added and stirred at rt for 24 h. The reaction mixture was quenched with aqueous NH ₄ Cl and the mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (1.36 g, 75%). ESI-MS m/z=436.08, 438.07 [MH] ^- .

Step 124b. To a suspension of the compound of step 124a (0.35 g, 0.80 mmol) and NMO (0.375 g, 3.2 mmol) in acetone-water (6 ml/1 ml) was added osmium tetroxide (1.0 ml, 0.080 mmol) and the mixture was stirred at rt for 18 h. It was quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO 3 and brine, dried over Na 2 SO 4 , filtered and concentrated to give a mixture of sulfone and sulfoxide.

Step 124c. To a solution of the compound of step 124b (156 mg, 0.32 mmol) in DMF (1.5 ml) at 0° C. are added NaH (45 mg, 60%, 1.12 mmol) and MeI (45 mg, 0.32 mmol ). After 1.5 h at 0°C, the reaction mixture is quenched with an aqueous solution of NH ₄ Cl, extracted with EtOAc, and the organic layer is washed with water and brine. The organic layer was dried (Na ₂ SO ₄ ), filtered, concentrated and purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid (61 mg, 38%). ESI-MS m/z=500.09, 502.09 [MH] ^- .

Step 124d. mCPBA (0.11 g, 77%, 0.49 mmol) was added to a solution of the compound of step 124c (61 mg, 0.122 mmol) in ΝΜΠ (1.5 ml) and stirred at rt for 20 h. Aqueous Na ₂ S was added. ₂ O ₃ , Na- 42 041670

HCO ₃ and a few drops of Et ₃ N and stirred at rt for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (63 mg, 100%). ESI-MS m/z=516.08, 518.08 [MH] ^- .

Example 125

Step 125a. In the solution of the compound of the stage prom. 3a (91 mg, 0.2 mmol) in DMF (1 mL) was added (S)-1-aminopropan-2-ol (45 mg, 0.6 mmol) and stirred at 90° C. for 20 h, then cooled. It is concentrated to give the desired crude compound, which is used in the next step without further purification. ESI-MS m/z=527.12, 529.12 [M-H]-.

Step 125b. To a solution of half of the compound of step 125a (~0.1 mmol) in CH ₂ Cl ₂ (1 ml) was added TEA (3 drops) and acetic anhydride (20 mg). The mixture is stirred for 2 hours and then concentrated. ΝΜΠ (1 mL) was added followed by m-CPBA (13.5 mg, 6 mmol) and stirred at rt for 20 h. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (18 mg, 30% in three steps). ESI-MS m/z=601.13, 603.13 [MH]-.

Example 130

Step 130a. To a solution of (R)-propane-1,2-diol (168 mg, 2.20 mmol) in DMF (3 ml) at 0°C was added NaH (88 mg, 60%, 2.20 mmol) and stirred at rt within 30 min. Add compound of step 34a (100 mg, 0.220 mmol) and heat to 55°C for 20 h. Cool to rt, quench with aqueous NH ₄ Cl, extract with EtOAc, wash with water and brine, dry over Na ₂ SO ₄ , filter, concentrated and purified by chromatography (silica, hexanes/EtOAc) to give the desired product (65 mg, 56%). ESI-MS m/z=528.12, 530.12 [MH]-.

Step 13OB. To a solution of the compound of step 130a (80 mg, 0.15 mmol) in ΝΜΠ (1.5 ml) was added m-CPBA (0.169 g, 77%, 0.75 mmol) and stirred at rt for 20 h. Add aqueous Na2S2O3, NaHCO3 and a few drops of Et ₃ N and stir at rt for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC with a C18 column and acetonitrile/ water as eluent to give the title compound (65 mg, 77%, single enantiomer). ESI-MS m/z=560.11, 562.11 [MH]-.

Example 136

The title compound (single enantiomer, structure determined experimentally) was isolated from the compound of Example 4 by SFC chromatography. ESI-MS m/z=546.10 [M+H]+.

Example 137

The title compound (single enantiomer, structure determined experimentally) was isolated from the compound of Example 5 by SFC chromatography. ESI-MS m/z=546.05 [M+H] ⁺ .

Example 141

Step 141a. To a solution of intermediate 1 (1.23 g, 3.88 mmol) and 3exo-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (801 mg, 3.52 mmol) in 50 ml of toluene is added 2-(tributyl-15-phosphanylidene)acetonitrile (2.13 g, 8.81 mmol), then stirred at 85°C in t/n. It is diluted with methyl tert-butyl ether, washed with an aqueous solution of 0.5 N NaOH, water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid.

- 43 041670 (1.63 g, 86%). ESI-MS m/z=525.12, 527.12 [MH] ^- .

Step 141b. To a solution of the compound of step 141a (1.31 g, 2.48 mmol) in N-methyl-2-pyrrolidinone (8.29 ml) at rt was added m-CPBA (1.95 g, 8.70 mmol), then the mixture kept at rt for 1 h. The reaction mixture was quenched with an aqueous solution of Na ₂ S ₂ O ₃ . The mixture is extracted with EtOAc and the combined organic phases are dried over Na2SO4, filtered and concentrated. The residue was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (1.28 g, 92%). ESI-MS m/z=557.11, 559.11 (M-H) ^- .

Step 141c. To a solution of the compound of step 141b (131 mg, 0.234 mmol) at rt was added 2 N HCl in dioxane, then the mixture was kept at rt for 4 h. The solution was concentrated to give a white solid (106 mg, 99%). ESI-MS m/z=457.06, 459.06 (M-H) ^- .

Step 141d. To a solution of the compound of step 141c (45 mg, 0.091 mmol) in DMF (1.8 mL) was added at rt i-Pr ₂ EtN (63.5 μL, 0.363 mmol), (methoxycarbonyl)-L-alanine (13.4 mg , 0.091 mmol) and HATU (51.8 mg, 0.136 mmol). The mixture is kept at rt overnight. The mixture is partitioned between EtOAc and water. The organic phase is washed with aqueous NaHCO3, water and brine, dried over Na2SO4, filtered and concentrated. The residue was purified by chromatography (silica, CH ₂ Cl ₂ /MeOH) to give the title compound as a white solid (37 mg, 69%, single enantiomer). ESI-MS m/z=586.10, 588.10 (M-H) ^- .

Example 144

To a solution of the compound of step 141c (49 mg, 0.099 mmol) in MeOH (0.99 ml) at rt was added (R)-2,2dimethyl-1,3-dioxolane-4-carbaldehyde (19.31 mg, 0.148 mmol) and NaCNBH ₄ (12.4 mg, 0.198 mmol) then the mixture is kept at rt overnight. The reaction mixture was quenched at rt with an aqueous solution of NH ₄ Cl. The reaction mixture is partitioned between EtOAc and water, then the organic phase is washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography (silica, CH ₂ Cl ₂ /MeOH) to give the title compound as a white solid (45 mg, 79%, single enantiomer). ESI-MS m/z=571.12, 573.12 (M-H) ^- .

Example 150

Step 150a. In the solution of the compound of the stage prom. 2e (246 g, 959 mmol) and pyridine (155 ml, 1.92 mol) in DCM (1 L) 3-nitrobenzenesulfonyl chloride (217 g, 978 mmol) was added at 0°C. The reaction mixture was stirred at 0°C for 30 min and at rt for 2 days. It is quenched with water (100 ml) and stirred at rt for 1 h, then extracted with MBTE (4 L). The organic layer was washed with water (2x1 L), 1 N HCl (1 L), water (500 ml), saturated NaHCO ₃ (500 ml) and brine (500 ml), dried over Na ₂ SO ₄ , filtered through celite and concentrated to about (500 ml), then add hexanes (500 ml). The mixture was concentrated in vacuo to initiate precipitation, cooled to rt, filtered and washed with cold hexanes to give the desired product (321 g, 76%).

Step 150b. To a solution of the compound of step 150a (108.6 g, 246 mmol) and intermediate 4 (70 g, 234 mmol) in DMF (250 ml) was added cesium carbonate (96 g, 295 mmol). The reaction mixture is degassed, heated slowly to 70°C and stirred at 70°C for 14 hours. It is cooled to rt, diluted with MBTE, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated to obtain the desired product (123 g , 98%), which is used without further purification.

Stage 150s. To a suspension of the compound of step 150b (50 g, 93 mmol) in MeOH (1.6 L) in an ice bath was added slowly concentrated HCl (200 ml) to keep the temperature below 30°C, then stirred at rt overnight. The reaction mixture is concentrated in vacuo to a volume of ~700 ml, cooled to 0°C, then filtered to collect a solid, which is washed with cold MeOH. The stock solution is then concentrated to 300 ml, cooled to 0° C. and filtered to collect a solid. The solid is dried in air for 14 hours to obtain the desired product (36 g, 91%).

Stage 150d. To a solution of the compound of step 150c (36 g, 85 mmol) in DMSO (150 ml) at rt was added IBX (30.9 g, 110 mmol), then stirred at 50° C. for 2 h. The mixture was poured into 1.2 L cold water, then extracted with EtOAc (2x500 ml). Some white solid is removed by filtration. The EtOAc extracts were washed with saturated aqueous NaHCO ₃ and brine and dried over Na ₂ SO ₄ . Filter and concentrate to ~150 ml, then cool to 0°C to obtain the desired solid, which is collected by filtration. The mother liquor is concentrated to 50 ml to obtain the second part of the target product (34.1 g, 95%).

- 44 041670

Stage 150e. To a solution of trimethylsulfoxonium iodide (36.3 g, 165 mmol) in DMF (180 ml) at 0°C was added potassium tert-butoxide (18.51 g, 165 mmol), then stirred at rt for 30 min. A solution of the compound of step 150d (43.5 g, 103 mmol) in DMF (120 ml) was added to the reaction mixture via cannula (temperature below 10°C) and stirred at rt for 16 h. The reaction mixture was poured into cold saturated NH ₄ Cl (500 ml) and MBTE (1.2 l). The organic layer was washed with water (3x500 ml) and saturated NaCl (2x300 ml). The organic layer is dried over Na ₂ SO ₄ , filtered through a pad of silica, washed with MBTE and concentrated to obtain the crude desired product, which is used without further purification.

Step 150f. To a solution of the crude material from step 150e in THF/water (220/70 ml) in an ice bath was added TFA (30.8 ml, 400 mmol), then stirred at rt for 6.5 h, then cooled to 0°C and slowly add 20 ml of saturated NaHCO ₃ , followed by the addition of solid NaOH (16.00 g, 400 mmol). Add EtOAc (800 ml) and water (600 ml). The organic layer was washed with a mixed solution of NaHCO3 and brine, then brine, dried (Na2SO4), filtered and concentrated to about 150 ml, then 450 ml of cyclohexane cooled to 0°C was added, filtered, washed with cyclohexane/EtOAc (3/1) to obtain target product (39 g, 83 mmol, 81% yield in 2 steps).

Stage 150g. To a solution of the compound of step 150f (5.75 g, 12.67 mmol) in ΝΜΠ (40 mL) at 0°C was added m-CPBA (8.52 g, 38.0 mmol, 77%) in one portion. Mixed in t / n at rt. Dilute with EtOAc, wash with Na ₂ S ₂ O ₃ , NaHCO ₃ , water and brine. Dry over Na ₂ SO ₄ , filter and concentrate. The crude product is recrystallized from MeOH. Dry at rt at t/n in vacuo to give the title compound (4.10 g, 8.44 mmol, 66.6% yield). ESI-MS m/z=484.08, 486.08 [MH] ^- .

Example 153

A solution of the compound of step 144a in AcOH (500 μl, 8.73 mmol) was stirred at rt overnight. The reaction mixture was concentrated and the residue was purified by chromatography (silica, CH ₂ Cl ₂ /MeOH) to give the title compound as a white solid (17 mg, 88%, single enantiomer). ESI-MS m/z=577.10, 579.10 (M+HCO2)-.

Example 156

Intermediate 3 (78 mg, 0.155 mmol) and sulfur diamide (44.7 mg, 0.465 mmol) in dioxane (0.5 ml) were stirred at 105°C for 3 h. The mixture was cooled to rt and purified by preparative HPLC using C18 column and acetonitrile/water as eluent to give the title compound (42 mg, 46%). ESI-MS m/z=580.06, 582.06 [MH] ^- .

Example 163

The two enantiomers of Example 36 are separated by chiral SFC. The title compound (pre-assigned) eluted earlier. ESI-MS m/z=590.11, 592.11 [MH] ^- .

Example 164

The two enantiomers of Example 36 are separated by chiral SFC. The title compound (pre-assigned) is eluted later. ESI-MS m/z=590.11, 592.11 [MH] ^- .

Example 174

The title compound (single enantiomer, preassigned, eluted earlier) was isolated from the compound of Example 82 by SFC chromatography. ESI-MS m/z=514.09, 516.09 [MH] ^- .

- 45 041670

The title compound (single enantiomer, preassigned, eluted later) was isolated from the compound of Example 82 by SFC chromatography. ESI-MS m/z=514.09,

516.09 [MH] ^- .

Example 180

Example 175

The title compound (single enantiomer, preassigned, eluted earlier) was isolated from Example 172 m/z=528.10, 530.10 [MH] ^- .

Example 181 using SFC chromatography. ESI-MS

The title compound (single enantiomer, preassigned, eluted later) was isolated from Example 172 m/z=528.10, 530.10 [MH] ^- .

Example 182 using SFC chromatography. ESI-MS

The title compound (single enantiomer, eluted earlier) was isolated from Example 173 by SFC chromatography. ESI-MS m/z=528.10, 530.10 [MH] ^- .

Example 183

The title compound (single enantiomer, eluted later) was isolated from Example 173 by SFC chromatography. ESI-MS m/z=528.10, 530.10 [MH] ^- .

Example 202

Step 202a. LDA (0.34 mmol ) freshly prepared in THF and stirred at the same temperature for 15 minutes, after which a solution of intermediate 2 (100 mg, 0.23 mmol) in THF (1 ml) was added. The temperature is raised to 0°C for 1 h and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, EtOAc/hexanes) to give the title compound (79 mg, contaminated with intermediate 2). ESI-MS m/z=598.12, 600.12 [MH] ^- .

Step 125b. To a solution of the compound of step 202a (79 mg, 0.13 mmol) in THF (2 ml) was added m-CPBA (112 mg, 0.5 mmol) and stirred at rt in ton. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (13 mg, 10% in two steps). ESIMS m/z=630.11, 632.11 [MH] ^- .

- 46 041670

Example 203

To a solution of Example 150 (207 mg, 0.426 mmol) in DMSO (2.1 ml) was added IBX (179 mg, 0.639 mmol). The reaction mixture was stirred at rt for 2 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (175 mg, 85%). ESI-MS m/z=481.6, 483.6 [MH]-.

Example 204

To a solution of the compound of Example 150 (103 mg, 0.212 mmol) in THF (2 ml) and NaHCO ₃ (aqueous saturated 1 ml) was added NaIO ₄ (140 mg, 0.655 mmol). The reaction mixture was stirred at rt for 2 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (60 mg, 62%). ESI-MS m/z=452.05, 454.05 [MH] ⁺ .

Example 205

To a solution of Example 203 (140 mg, 0.289 mmol), 2-methyl-2-butene (0.77 mL, 7.23 mmol), KH2PO4 (276 mg, 2.025 mmol) in THF (3 mL) and water (1 ml) add NaClO ₂ (294 mg, 2.60 mmol). The reaction mixture was stirred at rt for 1 h. The reaction mixture was acidified to pH 4 with 1 M HCl, then extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (109 mg, 75%). ESI-MS m/z=497.7, 499.6 [MH]-.

Example 206

To a solution of the compound of example 204 (22 mg, 0.048 mmol) in THF (0.5 ml) and MeOH (0.5 ml) was added NaBH ₄ (9 mg, 0.244 mmol). The reaction mixture was stirred at rt for 15 h. The reaction mixture was extracted with EtOAc, washed with water and brine. The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound as a white solid (21 mg, 95%, stereochemistry undetermined). ESI-MS m/z=453.6, 455.6 [MH]-.

Example 210

To a solution of the compound of step 11b (47 mg, 0.091 mmol) in THF (911 μl) at rt was added a solution of 2 N LiBH ₄ (22.77 μl, 0.046 mmol), then the mixture was kept at rt overnight. The reaction mixture was quenched with aqueous NH ₄ Cl and the mixture was extracted with EtOAc. The combined organic phases are washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography (silica, CH ₂ Cl ₂ /MeOH) to give the title compound as a white solid (14 mg, 30%). ESI-MS m/z=562.09, 564.09 (M+HCO2)-.

- 47 041670

Example 272

Step 272a. To a suspension of Me3SOI (1.98 g, 9.0 mmol) in DMF (5.0 ml) at 0°C was added t-BuOK (1.01 g, 9.0 mmol) and stirred at rt for 1 h. The reaction mixture is cooled to 0°C. The reaction mixture was charged dropwise with (1R,5S)-3-methylenebicyclo[3.2.1]octan-8-one-6,6,7,7-d4 in DMF (2.0 mL), which was prepared using the procedures similar to those described in the stage prom. 2a. The reaction mixture was stirred at rt for 2 h. 15% NH ₄ Cl was added dropwise to the reaction mixture and extracted with MTBE. The aqueous phase is re-extracted with MTBE. Then all organic phases are combined and washed with water and brine. The organic phase is dried over Na ₂ SO ₄ , filtered and concentrated to dryness to obtain the desired product (0.59 g, 64%).

Step 272b. To a solution of the compound of step 272a (0.59 g, 3.83 mmol) in THF/water (3.9/1.3 ml) at 0°C was added TFA (0.589 ml, 7.65 mmol). The reaction mixture is stirred at rt for 20 hours. The reaction mixture is cooled to 0° C. and Na ₂ CO ₃ is slowly added and adjusted to pH 7-8. Add EtOAc and water. The two layers are separated (brine is added to aid separation), the organic layer is washed with water and brine. The organic phase is dried over Na ₂ SO ₄ , filtered and concentrated to dryness to obtain the desired product (0.50 g, 76%).

Step 272c. To a solution of the compound of step 272b (0.48 g, 2.8 mmol) and imidazole (0.57 g, 8.4 mmol) in DMF (4.0 ml) was added TBSCl (0.63 g, 4.2 mmol) and stirred at rt for 18 hours. It is quenched by pouring into cold water. The product is extracted with hexanes. The organic phase was washed with water, dried over Na2SO4, filtered and concentrated to give the crude product (0.97 g, 100%).

Step 272d. To a solution of the compound of step 272c (1.43 g, 5.0 mmol) in dioxane-water (18/6.0 ml) was added 2,6-lutidine (1.16 ml, 10 mmol) and OsO ₄ (0.78 ml, 2.5% solution in t-butanol, 0.05 mmol). The mixture is cooled to 0° C. and NaIO ₄ (3.21 g, 15 mmol) is added. The suspension is stirred at rt for 16 h. An aqueous solution of Na2S2O3 is added. The mixture is stirred for 1 hour and filtered through celite. The mixture is extracted with MBTE/hexanes. The organic phase is washed with water, 1 N HCl, saturated NaHCO ₃ and brine. The organic phase is separated, dried over Na ₂ SO ₄ , filtered and concentrated to obtain the crude product (1.0 g, 69%).

Stage 272e. To a solution of LiBD ₄ (0.108 g, 4.2 mmol) at 0°C was added the compound of step 272d (0.577 g, 2.0 mmol) in MBTE (12 ml) dropwise. The resulting solution was stirred for 2 hours at 0°C. The reaction mixture is quenched by slow addition of aqueous NH4Cl keeping the temperature below 15°C. The reaction mixture is diluted with MBTE and water. The mixture is separated and the organic layer is washed with brine. The mixture is dried over Na2SO4, filtered and concentrated in vacuo. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.39 g, 67%).

Step 272f To a solution of the compound of step 272e (0.39 g, 1.34 mmol) and pyridine (0.216 ml, 2.68 mmol) in DCM (2 ml) at rt was added TsCl (0.306 g, 1.60 mmol) and stirred at rt for 40 h. It was quenched with H2O and extracted with EtOAc. The mixture was washed with H2O, 1 M HCl, saturated NaHCO3 and brine. The organic phase is collected, dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.59 g, 99%).

Stage 272g. To a solution of the compound of step 272f (1.7 g, 3.81 mmol) in MeOH (22 ml) was added concentrated HCl (1.0 ml) and stirred at rt for 16 h. It was diluted with EtOAc and the mixture was washed with water, saturated NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product is recrystallized from hexanes/MBTE to give the title compound (1.1 g, 87%).

Step 272h. To a solution of the compound of step 272g (0.16 g, 0.48 mmol), intermediate 6 (0.12 g, 0.40 mmol) and Cs ₂ CO ₃ (0.128 g, 0.392 mmol) in DMF (1.0 ml ) was stirred at 75° C. for 16 hours. It was diluted with EtOAc and the mixture was washed with water and brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.162 g, 88%). ESI-MS m/z=457.12, 459.12 [MH]'.

Step 272i. A solution of the compound of step 272h (0.16 g, 0.35 mmol) and m-CPBA (253 mg, 1.13 mmol, 77%) in ΝΜΠ (2.0 mL) was stirred at rt for 24 h. Aqueous Na was added ₂ S ₂ O ₃ , NaHCO ₃ and a few drops of Et3N and stirred at rt for 1 h. It was extracted with EtOAc, washed with water and brine, dried over Na2SO4, filtered and recrystallized from MeOH to give the title compound (118 mg, 68 %). ESI-MS m/z=489.11, 491.11 [MH] ^- .

- 48 041670

Example 279

DOH

Step 279a. To a suspension of Me3SOI (48.5 g, 220 mmol) in DMF (120 ml) at 0°C was added t-BuOK (24.72 g, 220 mmol) and stirred at rt for 1 h. The reaction mixture was cooled to 0° WITH. Intermediate 2a (20 g, 147 mmol) in DMF (80 ml) was added dropwise to the reaction mixture. The reaction mixture was stirred for 2 hours. 15% NH ₄ Cl was added dropwise to the reaction mixture and extracted with MTBE. The aqueous phase is re-extracted with MTBE. The combined organic layer was washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the desired product (19.9 g, 90%).

Step 279b. TFA (30.8 ml, 399 mmol) is added to a solution of the compound of step 279a (20 g, 133 mmol) in THF/water (150/50 ml) at 0°C. The reaction mixture is stirred at rt for 3 h, then cooled to 0°C, Na ₂ CO ₃ is slowly added and adjusted to pH 7-8. Add EtOAc and water. The two layers are separated (brine is added to aid separation), the organic layer is washed with water and brine, dried over Na2SO4, filtered and concentrated to give the desired product (17.7 g, 79%).

Step 279c. To a solution of the compound of step 279b (10 g, 59.4 mmol) and imidazole (10.2 g, 148 mmol) in DMF (80 ml) was added TBSCl (10.7 g, 71.3 mmol) and stirred at rt for 6 pm It is quenched by pouring into cold water. The product is extracted with hexanes. The organic phase is washed with water, dried over Na ₂ SO ₄ , filtered and concentrated to obtain the crude product (18.1 g, 100%).

Stage 279d. To a solution of the compound of step 279c (16.8 g, 59.4 mmol) in dioxane-water (170/60 ml) was added 2,6-lutidine (13.8 ml, 119 mmol) and OsO ₄ (9.3 ml, 2.5% solution in t-butanol, 0.59 mmol). The mixture is cooled and NaIO ₄ (37.1 g, 178.2 mmol) is added. The suspension is stirred at rt for 16 h. An aqueous solution of Na ₂ S ₂ O ₃ is added. The mixture is stirred for 1 hour and filtered through celite. The mixture is extracted with MBTE/hexanes. The organic phase is washed with water, 1N HCl, saturated NaHCO3 and brine. The organic phase is separated, dried over Na2SO4, filtered and concentrated to give the crude product (16.6 g, 94%).

Stage 279f. To a solution of the compound of step 279d (143 mg, 0.503 mmol) in CD3OD (3.0 mL) was added MeONa (5.4 mg, 0.10 mmol) and the mixture was stirred at rt for 3 h. dissolved in CD ₃ OD (3.0 ml) and stirred for ₃ h. ). After 1 h, it is quenched by the slow addition of aqueous NH4Cl. Add MBTE and water. The mixture is separated and the organic layer is washed with brine. The mixture is dried over Na2SO4, filtered and concentrated in vacuo. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.070 g, 50%).

Stage 279g. To a mixture of intermediate 6 (73 mg, 0.24 mmol) and the compounds of step 279f (71 mg, 0.24 mmol) in toluene (1.5 ml) at rt was added cyanomethylenetributylphosphorane (176 mg, 0.73 mmol) and stirred at 75° C. for 16 hours. The mixture was cooled to rt and purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.11 g, 79%). ESI-MS m/z=571.21, 573.21 [MH] ^- .

Step 279h. To a solution of the compound of step 279g (0.11 g, 0.19 mmol) in MeOH (3.0 ml) was added concentrated HCl (0.3 ml) and stirred at rt for 2 h. It was diluted with EtOAc and the mixture was washed with water saturated with NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was recrystallized from hexanes/MBTE to give the title compound (0.083 g, 94%). ESI-MS m/z=457.12, 459.12 [MH] ^- .

Stage 279i. A solution of the compound of step 279h (0.083 g, 0.18 mmol) and m-CPBA (0.14 mg, 0.63 mmol, 77%) in NMP (2.0 mL) was stirred at rt for 24 h. Aqueous Na was added ₂ S ₂ O ₃ , NaHCO ₃ and a few drops of Et ₃ N and stirred at rt for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC (C18 and acetonitrile /water) to obtain the title compound (48 mg, 54%). ESI-MS m/z=489.11.491.11 [MH] ^- .

Example 281

Step 281a. To a solution of the compound of step 150f (4.0 g, 8.11 mmol) in DMSO (22 mL) was added IBX (3.70 g, 13.22 mmol) at rt. The reaction mixture is heated to 50°C for 3 hours Then the reaction mixture is cooled to rt and diluted with EtOAc. The reaction mixture is filtered, the solid is discarded, and the filtrate is washed with water and saturated NaCl. The organic layer was dried with Na ₂ SO ₄ , filtered and concentrated to give the crude desired product, which was used without further purification (3.90 g, 98%).

Step 281b. To a solution of the compound of step 281a (50 mg, 0.111 mmol) in THF (0.53 ml) at -78°C was added a complex of isoropyl magnesium chloride and lithium chloride (237 μl, 0.243 mmol, 1.3 M in THF). The reaction mixture is stirred for 30 min and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, EtOAc/hexanes) to give the title compound (18 mg, 33%). ESI-MS m/z=540.14, 542.14 [M+HCO2]-.

Step 281c. To a solution of the compound of step 281b (18 mg, 0.01 mmol) in ΝΜΠ (0.5 ml) was added m-CPBA (24 mg, 0.096 mmol, 77%) and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid [M+HCO2]-.

Example 283 colors (14 mg, 73%). ESI-MS m/z=572.13, 574.13

Step 283a. To a solution of the compound of step 281a (50 mg, 0.111 mmol) in THF (0.53 ml) at -78°C was added phenylmagnesium bromide (237 μl, 0.243 mmol, 1 M in THF). The reaction mixture is stirred for 30 min and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, EtOAc/hexanes) to give the title compound (17 mg, 29%). ESI-MS m/z=574.13, 576.13 [M+HCO2]-.

Step 283b. To a solution of the compound of step 283a (17 mg, 0.03 mmol) in ΝΜΠ (0.5 ml) was added m-CPBA (22 mg, 0.096 mmol, 77%) and stirred at rt in t/n. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (3 mg, 17%). ESI-MS m/z=606.12, 608.12 [M+HCO2]-.

Example 284

Step 284a. To a solution of 2-bromopyridine (46 μl, 0.49 mmol) in THF (1 ml) at 0°C was added a complex of isoropyl magnesium chloride and lithium chloride (0.49 ml, 0.49 mmol, 1.3 M in THF). The reaction mixture is heated to rt and stirred for 30 min, then cooled to -78°C followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture is stirred for 18 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (11 mg, 9%). ESI-MS m/z=575.12, 577.12 [M+HCO2]-.

Step 284b. To a solution of the compound of step 284a (11 mg, 0.02 mmol) in ΝΜΠ (1 mL) was added p-TSA (20 mg, 0.105 mmol) and m-CPBA (20 mg, 0.089 mmol, 77%) and stirred at rt in t / n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (6 mg, 51%). ESIMS m/z=607.13, 609.13 [M+HCO2].

Example 284a he

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 284 by SFC chromatography, previously eluting the compound.

- 50 041670 he

A^

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 284 by SFC chromatography, eluting the compound later.

Example 285

Example 284b he

A^'

Step 285a. To a solution of 3-bromopyridine (46 μl, 0.49 mmol) in THF (1 ml) at 0°C was added a complex of isoropyl magnesium chloride and lithium chloride (0.49 ml, 0.49 mmol, 1.3 M in THF). The reaction mixture is heated to rt and stirred for 30 min, then cooled to -78°C followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture is stirred for 18 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (15 mg, 13%). ESI-MS m/z=575.12, 577.12 [M+HCO2] ^- .

Step 285b. To a solution of the compound of step 285a (15 mg, 0.028 mmol) in ΝΜΠ (1 ml) was added p-TSA (20 mg, 0.105 mmol) and m-CPBA (20 mg, 0.089 mmol, 77%) and stirred at rt at t /n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (8 mg, 50%). ESI-MS m/z=607.13, 609.13 [M+HCO2] ^- .

Example 285a he

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 285 by SFC chromatography, previously eluting the compound.

Example 285b he

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 285 by SFC chromatography, eluting the compound later.

Example 286

Step 286a. To a solution of 4-iodopyridine (159 mg, 0.774 mmol) in THF (1.5 ml) at 0°C was added n-butyllithium (0.31 ml, 0.774 mmol, 2.5 M in hexanes). This reaction mixture is heated to rt and stirred for 30 min, then cooled to -78°C followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture was stirred for 18 h, warming slowly to rt, and quenched with aqueous NH4Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (45 mg, 38%). ESI-MS m/z 575.30, 577.30 [M+HCO2] ^- .

Step 286b. To a solution of the compound of step 286a (45 mg, 0.085 mmol) in ΝΜΠ (2 ml) was added p-TSA (48 mg, 0.254 mmol) and m-CPBA (57 mg, 0.254 mmol, 77%) and stirred at rt at t /n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (8 mg, 17%). ESI-MS m/z=607.29, 609.29 [M+HCO2] ^- .

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Example 287

Step 287a. To a solution of 1-(diethoxy)-1H-imidazole (159 mg, 0.487 mmol) in THF (1 ml) at -78°C was added n-butyllithium (0.195 ml, 0.487 mmol, 2.5 M in hexanes). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture is stirred for 18 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (22 mg, 38%). ESI-MS m/z=564.29, 566.29 [M+HCO2] ^- .

Step 287b. To a solution of the compound of step 287a (22 mg, 0.042 mmol) in NMn (2 ml) was added p-TSA (50 mg, 0.263 mmol) and m-CPBA (50 mg, 0.223 mmol, 77%) and stirred at rt at t /n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound as a white solid (6 mg, 26%). ESI-MS m/z=596.28, 598.28 [M+HCO2] ^- .

Example 288

Step 288a. To a solution of 1-methyl-1H-pyrazole (64 μl, 0.774 mmol) in THF (1.5 ml) at 0°C was added n-butyllithium (0.31 ml, 0.774 mmol, 2.5 M in hexanes). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture is stirred for 18 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (79 mg, 67%). ESI-MS m/z=578.32, 580.31 [M+HCO2] ^- .

Step 288b. To a solution of the compound of step 288a (79 mg, 0.148 mmol) in ΝΜΠ (2 ml) was added m-CPBA (99 mg, 0.444 mmol, 77%) and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, acetone/hexane) to give the title compound as a white solid (65 mg, 78%). ESI-MS m/z=610.30, 612.30 [M+HCO2] ^- .

Example 289

Step 289a. To a solution of 4-bromo-1-methyl-1H-pyrazole (69 μl, 0.664 mmol) in THF (2 ml) at -78°C was added n-butyllithium (0.266 ml, 0.664 mmol, 2.5 M in hexanes). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (100 mg, 0.221 mmol) in THF (1 ml). The reaction mixture was stirred for 18 h, warming slowly to rt, and quenched with aqueous NH4Cl. It is extracted with EtOAc, washed with brine and dried (Na2SO4). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (23 mg, 19%). ESI-MS m/z=578.31, 580.31 [M+HCO ₂ ] ^- .

Step 289b. To a solution of the compound of step 289a (23 mg, 0.148 mmol) in ΝΜΠ (2 mL) was added m-CPBA (29 mg, 0.444 mmol, 77%) and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound as a white solid (22 mg, 90%). ESI-MS m/z=610.30, 612.31 [M+HCO2] ^- .

Example 290

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Step 290a. To a solution of oxazole (76 μl, 1.162 mmol) in THF (1.5 ml) at 0°C was added i-PrMgClLiCl (0.894 ml, 1.162 mmol, 1.3 M in THF). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (150 mg, 0.332 mmol) in THF (1.5 ml). The reaction mixture is stirred for 3 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na2SO4). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (22 mg, 38%). ESI-MS m/z=565.28, 567.28 [M+HCO ₂ ] ^- .

Step 290b. To a solution of the compound of step 290a (133 mg, 0.042 mmol) in NMn (2.5 ml) was added rTSA (243 mg, 1.276 mmol) and m-CPBA (172 mg, 0.766 mmol, 77%) and stirred at rt at t /n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to afford the title compound as a white solid (20 mg, 14%). ESI-MS m/z=597.26, 599.26 [M+HCO2] ^- .

Example 291

Step 291a. To a solution of 2-bromo-5-fluoropyridine (302 mg, 1.715 mmol) in THF (3 ml) at -78°C was added n-butyllithium (0.730 ml, 1.826 mmol, 2.5 M in THF) dropwise. After stirring for 1 hour at the same temperature, a solution of the compound of step 281a (250 mg, 0.553 mmol) in THF (1.5 ml) was added to the mixture at -78°C. The reaction mixture is slowly warmed to rt and stirred for 16 hours Add a saturated solution of NH ₄ Cl. The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica, MeOH/DCM) to give the title compound (70 mg, 0.128 mmol, 23% yield) as a pale brown solid.

Step 291b. To a solution of the compound of step 291a (70 mg, 0.128 mmol) and p-TSA (72.8 mg, 0.383 mmol) in ΝΜΠ (1 ml) at 0°C was added m-CPBA (86 mg, 0.383 mmol, 77%). The reaction mixture is slowly heated to rt and stirred for 16 h. Na ₂ S ₂ O ₃ solution and NaHCO ₃ solution are added and the resulting mixture is stirred at rt for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution , NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, MeOH/DCM) to give the title compound (17.0 mg, 0.128 mmol, 23% yield) as a light brown solid. ESI-MS m/z=580.00, 582.01 [MH] ^- .

Example 291a

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 291 by SFC chromatography, previously eluting the compound.

Example 291b he

The title compound (single enantiomer, provisionally assigned) was isolated from the compound of Example 291 by SFC chromatography, eluting the compound later.

Example 292

The crude material of step 291b was purified by column chromatography (0-20% MeOH in DCM) to give the title compound (9.3 mg, 0.016 mmol, 12% yield) as a light brown solid. ESI-MS m/z=596.00, 598.01 [MH] ^- .

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Example 293 he

Step 293a. To a solution of 2-bromo-5-methylpyridine (200 mg, 1.16 mmol) in THF (1.5 ml) at -78°C was added n-butyl lithium (0.465 ml, 1.16 mmol, 2.5 M in hexane ). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (150 mg, 0.332 mmol) in THF (1.5 ml). The reaction mixture is stirred for 1 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (129 mg, 71%). ESI-MS m/z=589.14, 591.14 [M+HCO2] ^- .

Step 293b. To a solution of the compound of step 293a (129 mg, 0.24 mmol) in ΝΜΠ (2.4 ml) was added p-TSA (225 mg, 1.18 mmol) and m-CPBA (159 mg, 0.71 mmol, 77% ), and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (109 mg, 80%). ESI-MS m/z=621.13, 623.13 [M+HCO2] ^- .

Example 297

Step 297a. To a solution of 2-bromo-3-fluoropyridine (204 mg, 1.16 mmol) in THF (1.5 ml) at -78°C was added n-butyllithium (0.465 ml, 1.16 mmol, 2.5 M in hexane ). The reaction mixture was stirred for 30 minutes followed by the addition of a solution of the compound of step 281a (150 mg, 0.332 mmol) in THF (1.5 ml). The reaction mixture is stirred for 1 h, warming slowly to rt, and quenched with aqueous NH ₄ Cl. It is extracted with EtOAc, washed with brine and dried (Na ₂ SO ₄ ). After concentration, the crude product was purified by chromatography (silica, acetone/hexanes) to give the title compound (10 mg, 5.5%). ESI-MS m/z=563.12, 565.10 [M+HCO2] ^- . 3-(((1R,3r,5S,8r)-8-((2-bromo-3fluoropyridin-4-yl)(hydroxu)metul)-8-hydroxubucyclo[3.2.1]octan-3-yl) is also isolated thio)-4-chloro-N-(3,4difluorophenyl)benzamide (77 mg, 37%). ESI-MS m/z=671.03, 673.04 [M+HCO2] ^- .

Step 297b. To a solution of the compound of step 297a (10 mg, 0.018 mmol) in NMn (0.2 mL) was added p-TSA (17 mg, 0.09 mmol) and m-CPBA (12 mg, 0.055 mmol, 77%) and stirred at ct in t / n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (4.7 mg, 44%). ESI-MS m/z=625.11, 627.11 [M+HCO2] ^- .

Example 298 f Ϊ ^Η ίΓ^Τ

TI S _ SoCh ^{he f}

Step 298. To the solution of the by-product of Step 297a (3-(((1R,3r,5S,8r)-8-((2-bromo-3-fluoropyridin-4yl)(hydroxy)methyl)-8-hydroxybicyclo[3.2. 1]octan-3-yl)thio)-4-chloro-N-(3,4-difluorophenyl)benzamide, 77 mg, 0.123 mmol) in NMn (1.3 ml) add p-TSA (117 mg, 0.613 mmol ) and m-CPBA (82 mg, 0.368 mmol, 77%), and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na2SO4), filtered and concentrated. The crude product was purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid (64 mg, 79%). ESI-MS m/z=703.01, 705.12 [M+HCO2] ^- .

Example 307

Step 307a. Nonafluorobutane-1-sulfonyl fluoride ( 5.52 g, 18.27 mmol) and stirred at this temperature for 1 hour. Add 200 ml of

- 54 041670

MTBE. The solution was washed with water, twice with 1N HCl, then with water, twice with aqueous NaHCO3 and twice with brine. After drying (Na2SO4) and concentration, 13.5 g of a yellow solid are obtained. MTBE/hexanes ~1:10 (200 ml) were added to it and stirred at rt for 30 min, after which it was filtered under vacuum. The collected solid was washed with MTBE/hexanes (1:10, 100 mL) to give an off-white solid (10.1 g, 86%). ESI-MS m/z=704.02, 706.02 [MH] ^- .

Step 307b. Methylisopropylamine (0.15 ml) was added to a solution of the compound of step 307a (200 mg, 0.283 mmol) in DMF (2 ml) and stirred at 75° C. for two days. The crude product is diluted with EtOAc, washed twice with water, washed twice with brine, dried (Na ₂ SO ₄ ) and concentrated. The crude product is purified by chromatography (silica, MeOH/DCM) to give the title compound as a mixture with other impurities. ESI-MS m/z=477.15, 479.15 [MH]-.

Step 307c. The title compound was prepared from the compound of step 307b by the procedure described in Example 284b and purified by preparative HPLC. ESI-MS m/z=509.14, 511.14 [M-H]-.

Example 308 he and

F'^4^

H°

To a solution of step 82a (570 mg, 1.27 mmol) in THF (10 mL) was added m-CPBA (77 w/w%, 1.42 g, 6.33 mmol) and stirred at rt in t/n . Aqueous Na ₂ S ₂ O ₃ (10 ml) and NaHCO ₃ (10 ml) were added thereto, followed by 4 drops of TEA and EtOAc (50 ml). It is stirred at rt for 1.5 h before separation. The aqueous phase is extracted with EtOAc. The combined organic phase was washed with NaHCO ₃ and brine and dried (Na ₂ SO ₄ ). After concentration, the crude pale yellow solid was dissolved in MeOH (50 ml) with heating (air jet) and slowly cooled to rt, after which it was kept at 0°C for 30 min. The resulting crystals were collected in vacuo to give the title compound (510 mg, 81%) as a white solid. ESI-MS m/z=496.20, 498.20 [MH] ^- .

Example 309

To a solution of Example 308 (50 mg, 0.10 mmol) in DMF (0.5 mL) was added 1,1dioxoisothiazolidine (18.3 mg, 0.15 mmol) followed by K2CO3 (14 mg, 0.1 mmol) and stirred at 45°C in t/n. The crude product was purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid (16 mg, 26%). ESI-MS m/z=617.25, 619.25 [M-H]-.

Example 314 he

Step 314a. To a suspension of Me3SOI (365 mg, 1.66 mmol) in DMF (1 ml) at 0°C was added potassium tert-butoxide (186 mg, 1.66 mmol). The mixture is heated to rt and stirred for 45 min. The compound of step 281a (150 mg, 0.33 mmol) in DMF (2 mL) was added dropwise to the reaction mixture. After stirring for 4 h at rt, the reaction mixture is cooled to 0°C, a saturated solution of NH ₄ Cl is added, extracted with MTBE and dried over Na ₂ SO ₄ . The crude product was purified by column chromatography (0-50% EtOAc in hexane) to give the title compound (42.0 mg, 0.090 mmol, 27% yield) as a white solid.

Step 314b. To a solution of the compound of step 314a (42 mg, 0.090 mmol) in DMF (2 mL) was added Spirazol (12.27 mg, 0.180 mmol) and potassium carbonate (18.69 mg, 0.135 mmol). The reaction mixture was stirred at 65° C. for 15 hours. The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (0-70% EtOAc in hexane) to obtain the target compound (30.0 mg, 0.056 mmol, 62% yield ) as a whitish solid.

Step 314c. m-CPBA (37.8 mg , 0.169 mmol, 77%). The reaction mixture is slowly heated to rt and stirred for 15 h. Saturated Na ₂ S ₂ O ₃ solution and NaHCO ₃ solution are added to the mixture and stirred for 1 h. The reaction mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution, NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (20 mg, 0.035 mmol, 63% yield) as an off-white solid. ESI-MS m/z=565.30, 567.29 [MH]-.

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0.483 mmol) in DMF (1 ml) at 0° C. is added

Example 315

Step 315a. To a solution of oxazolidin-2-one (42.0 mg, sodium hydride (17.38 mg, 0.435 mmol). The mixture was stirred for 1 h at rt and cooled to 0°C. The compound of step 314a (45 mg , 0.097 mmol) in DMF (1 ml) The mixture is heated to 55°C for 15 hours The reaction mixture is cooled to rt and a saturated solution of NH4Cl is added The mixture is extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica , hexanes/EtOAc) to give the title compound (40 mg, 0.072 mmol, 75% yield) as a white solid.

Step 315b. To a solution of the compound of step 315a (38 mg, 0.069 mmol) in ΝΜΠ (1 ml) at 0°C was added m-CPBA (46.2 mg, 0.206 mmol, 77%). The reaction mixture is slowly heated to rt and stirred for 16 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution, NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, MeOH/DCM) to give the title compound (14.0 mg, 0.024 mmol, 35% yield) as an off-white solid. ESI-MS m/z=584.02, 586.02 [MH]'.

Example 316

Step 316a. To a solution of 1-(diethoxymethyl)-1H-imidazole (0.053 mg, 0.322 mmol) in THF (2 mL) at 78°C was added n-butyl lithium (0.116 mL, 0.290 mmol, 2.5 M in THF). The mixture is heated to 0°C and stirred for 45 minutes and then cooled to -78°C. The compound of step 314a (30 mg, 0.064 mmol) in THF (1 mL) was added to the reaction mixture. The reaction mixture is heated to 50° C. and stirred for 16 h. The reaction mixture is cooled to rt and a saturated solution of NH ₄ Cl is added. The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica, hexanes/acetone) to give the title compound (13 mg, 0.024 mmol, 38% yield) as an off-white solid.

Step 316b. m-CPBA (16.37 mg, 0.073 mmol, 77%) is added to a solution of the compound of step 316a (13 mg, 0.024 mmol) and KSK (8.48 mg, 0.037 mmol) in ΝΜΠ (1 ml) at 0°C. The reaction mixture is slowly heated to rt and stirred for 15 h. The mixture is extracted with EtOAc, washed with saturated Na2S2O3 solution, NaHCO3 solution (x2) and brine, and dried over Na2SO4. The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (5.8 mg, 0.010 mmol, 42% yield) as an off-white solid. ESIMS m/z=565.02, 566.02 [MH] ^- .

Example 317

Step 317a. In a solution of 4-methylpyridine (0.134 ml,

1.372 mmol) in THF (3 ml) at -78° C., lithium diisopropylamide (1.372 ml, 1.372 mmol, 1 M in THF) is added dropwise. The mixture is stirred for 1 hour at the same temperature. A solution of the compound of step 281a (200 mg, 0.443 mmol) in THF (1.5 ml) was added to the mixture at -78°C. The reaction mixture is slowly heated to rt and stirred for 16 h. Saturated NH4C1 solution is added. The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica, MeOH/DCM) to give the title compound (91 mg, 0.167 mmol, 38% yield) as a sticky oil.

Step 317b. m-CPBA (112 mg, 0.501 mmol, 77%) was added to a solution of the compound of step 317a (91 mg, 0.167 mmol) and p-TSA (95 mg, 0.501 mmol) in ΝΜΠ (1 ml) at 0°C. The reaction mixture is slowly heated to rt and stirred for 16 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution, NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (61.0 mg, 0.106 mmol, 63% yield) as a white solid. ESI-MS m/z=576.04, 578.04 [MH] ^- .

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1.372 mmol) in THF (3 ml) at -78°C dropwise

Example 318

Step 318a. To a solution of 3-methylpyridine (0.134 mL, lithium diisopropylamide (1.372 mL, 1.372 mmol, 1 M in THF) was added. After stirring for 30 min at the same temperature, the mixture was heated to 0°C, stirred for 30 min, and cooled to -78° C. A solution of the compound of step 281a (200 mg, 0.443 mmol) in THF (1.5 ml) is added to the mixture at -78° C. The reaction mixture is slowly heated to rt and stirred for 15 h. Saturated NH solution is added ₄ Cl The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica, MeOH/DCM) to give the title compound (70 mg, 0.128 mmol, 29% yield) as a pale yellow solid.

Step 318b. To a solution of the compound of step 318a (70 mg, 0.128 mmol) and p-TSA (73.3 mg, 0.385 mmol) in ΝΜΠ (1 ml) at 0°C was added m-CPBA (86 mg, 0.385 mmol, 77%). The reaction mixture is slowly heated to rt and stirred for 16 h. Saturated Na ₂ S ₂ O ₃ solution and NaHCO ₃ solution are added and the resulting mixture is stirred at rt for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O solution ₃ , NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (27.0 mg, 0.047 mmol, 36% yield) as a white solid. ESI-MS m/z=576.04, 578.04 [MH] ^- .

Example 319

Step 319a. To a solution of 2-methylpyridine (0.135 ml, 1.372 mmol) in THF (3 ml) at -78° C., lithium diisopropylamide (0.549 ml, 1.372 mmol, 1 M in THF) was added dropwise. After stirring for 1 hour at the same temperature, a solution of the compound of step 281a (200 mg, 0.443 mmol) in THF (1.5 ml) was added to the mixture at -78°C. The reaction mixture is slowly warmed to rt and stirred for 16 hours Add a saturated solution of NH ₄ Cl. The mixture was extracted with EtOAc, dried over Na ₂ SO ₄ and purified by column chromatography (silica, MeOH/DCM) to give the title compound (130 mg, 0.239 mmol, 54% yield) as a pale yellow solid.

Step 319b. To a solution of the compound of step 319a (130 mg, 0.239 mmol) and p-TSA (136 mg, 0.716 mmol) in ΝΜΠ (1 ml) at 0°C was added m-CPBA (160 mg, 0.716 mmol, 77%). The reaction mixture is slowly heated to rt and stirred for 16 h. Saturated Na ₂ S ₂ O ₃ solution and NaHCO3 solution are added and the resulting mixture is stirred at rt for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution , NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (80.0 mg, 0.139 mmol, 58% yield) as a white solid. ESI-MS m/z=576.04, 578.04 [MH] ^- .

Example 320

Step 320a. To a solution of Example 82a (450 mg, 1.00 mmol) in DMF (5 mL) was added allyl bromide (363 mg, 3.0 mmol) followed by the second generation Hoveid-Grubbs catalyst (12.5 mg, 0.2 mmol). ) and stirred at rt in t/n. The reaction mixture is concentrated. The crude product was purified by chromatography (silica, EtOAc/hexanes) to give the title compound (410 mg, 1.1:1 mixture of target and starting materials) as an off-white solid. ESI-MS m/z=542.16, 544.16 [MH] ^- .

Step 320b. To a solution of the compound of step 320a (170 mg, 0.31 mmol) in DMF (2 mL) was added dioxoisothiazolidine (38 mg, 0.31 mmol) followed by K ₂ CO ₃ (44 mg, 0.31 mmol) and stirred at 65°С in t/n. The crude product is diluted with EtOAc, washed twice with water, washed twice with brine, dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by chromatography (silica, MTBE/hexanes) to give the title compound (59 mg, 32%) as an off-white solid. ESI-MS m/z=627.12, 629.12 [Mn+nsoon] ^- .

Step 320c. The title compound was prepared from the compound of step 320b following the procedure described in Example 4b. ESI-MS m/z=693.30, 695.30 [M-H+HCOOH] ^- .

- 57 041670

Example 321

Step 321a. To a solution of the compound of step 82a (100 mg, 0.222 mmol) and 3-bromopyridine (26 μl, 0.267 mmol) in DMF (2 ml) was added Et ₃ N (62 μl, 0.445 mmol), mg, 0.022 mmol) and palladium acetate (2.5 mg, 0.011 mmol). The reaction mixture is stirred at 120° C. for 18 hours. The reaction mixture is cooled, then extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by column chromatography (silica, hexane/acetone) to give the title compound (100 mg, 0.190 mmol, 85% yield). ESI-MS m/z=571.30, 573.30 [MH] ^- .

Step 321b. To a solution of the compound of step 321a (52 mg, 0.099 mmol) in acetone (1 ml) and water (0.2 ml) was added OsO ₄ (25 μl, 4.93 μmol, 5% in water) and NMO (58 mg, 0.493 mmol). The reaction mixture is stirred at rt for 72 hours. The reaction mixture is diluted with ethyl acetate, then washed with saturated Na ₂ S ₂ O ₃ and saturated NaCl. The organic layer is dried with Na ₂ SO ₄ , filtered and concentrated. The crude material was used without further purification.

Step 321c. To a solution of the crude material from step 321b in ΝΜΠ (2 ml) was added p-TSA (56 mg, 0.294 mmol) and m-CPBA (110 mg, 0.49 mmol, 77%) and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product is purified by chromatography (silica, hexane/acetone) to give the title compound at m/z=637.32, 639.32 [M+HCO ₂ ] ^- .

Example 322 as a white solid. ESI-MS

Step 322a. To a solution of the compound of step 82a (100 mg, 0.222 mmol) and 4-bromo-1-methyl-1H-pyrazole (28 μl, 0.267 mmol) in DMF (2 ml) was added Et ₃ N (62 μl, 0.445 mmol), three -o-tolylphosphine (6.8 mg, 0.022 mmol) and palladium acetate (2.5 mg, 0.011 mmol). The reaction mixture is stirred at 120° C. for 18 hours. The reaction mixture is cooled and extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by column chromatography (silica, hexane/acetone) to give the title compound (12 mg, 0.023 mmol, 10% yield). ESI-MS m/z=574.32, 576.32 [MH] ^- .

Step 322b. To a solution of the compound of step 322a (12 mg, 0.099 mmol) in acetone (0.5 ml) and water (0.1 ml) was added OsO ₄ (6 μl, 1.13 μmol, 5% in water) and NMO (13 mg , 0.113 mmol). The reaction mixture was stirred at rt for 72 h. The reaction mixture was diluted with ethyl acetate and then washed with saturated Na2S2O3 and saturated NaCl. The organic layer is dried with Na2SO4, filtered and concentrated. The crude material was used without further purification.

Step 322c. To a solution of the crude material from step 322b in ΝΜΠ (1 ml) was added p-CPBA (16 mg, 0.069 mmol, 77%) and stirred at rt in t/n. The reaction mixture is extracted with EtOAc, washed with water and brine. The organic layer is dried (Na ₂ SO ₄ ), filtered and concentrated. The crude product is purified by chromatography (silica, hexane/acetone) to give the title compound as a white solid. ESI-MS m/z=640.33, 642.33 [M+HCO ₂ ] ^- .

Example 323

Step 323a. To a solution of trimethylsilylacetylene (0.50 ml, 3.56 mmol) in THF (1.66 ml) at -78°C was added n-BuLi (1.426 ml, 3.56 mmol, 2.5 M in THF). The resulting mixture was stirred for 1 hour at the same temperature. A solution of the compound of step 281a (0.3 g, 0.664 mmol) in THF (1.66 ml) was added to the reaction mixture at -78°C. The light yellow solution obtained is heated slowly to rt and stirred for 3 h. Saturated NH ₄ Cl solution is added. The mixture is extracted with EtOAc, dried over Na ₂ SO ₄ and used without further purification.

Step 323b. Potassium carbonate (459 mg, 3.32 mmol) was added to a solution of the compound of step 323a (365 mg, 0.664 mmol) in MeOH (3 ml) and water (0.3 ml) at rt. The mixture is stirred for 15 h at rt. The solvent is removed in vacuo, the resulting mixture is extracted with EtOAc, dried over Na ₂ SO ₄ and then purified by column chromatography (silica, hexanes/EtOAc) to obtain the target compound

- 58 041670 (204.9 mg, 0.429 mmol, 65% yield) as a white solid.

Step 323c. A mixture of formaldehyde (109 µl, 1.465 mmol), acetic acid (12.58 µl, 0.220 mmol) and THF (1 ml) was stirred for 15 min at rt. Sodium azide (14.28 mg, 0.220 mmol) is added followed by the compound of step 323b (70 mg, 0.146 mmol). The mixture is stirred for 10 minutes, sodium ascorbate (5.80 mg, 0.029 mmol) is added followed by copper(II) sulfate (1.169 mg, 7.32 µmol) in 150 µl of water. The reaction mixture was heated to 50° C. and stirred for 16 h, and then extracted with EtOAc and dried over Na ₂ SO ₄ and then purified by column chromatography (silica, MeOH/DCM) to give the target compound (53.4 mg, 0.103 mmol, 70% yield) as a greenish solid.

Stage 323d. m-CPBA (68.9 mg, 0.307 mmol, 77% ). The reaction mixture is slowly heated to rt and stirred for 15 h. Saturated Na ₂ S ₂ O ₃ solution and NaHCO ₃ solution are added and the resulting mixture is stirred at rt for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O solution ₃ , NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, MeOH/DCM with 0.1% NH ₃ ) to give the title compound (15.0 mg, 0.027 mmol, 27% yield) as an off-white solid. ESI-MS m/z=549.95, 551.97 [MH] ^- .

Example 327

Step 327a. Processing the compound of step 279d under the conditions described in step prom. 2e gives the target product. ¹ H-NMR (400 MHz, chloroform-d) δ 3.90 (m, 1H), 3.43 (s, 2H), 2.69 (s, 1H), 1.84 (m, 4H), 1 .70 (m 2H), 1.57 (m, 2H), 1.45 (t, J=6.7 Hz, 3H), 0.83 (s, 9H), 0.00 (s, 6H).

Step 327b. Pyridine ( _8.98 ml, ₁₁₁ mmol), DMAP (0.226 g, 1.850 mmol ) and benzenesulfonyl chloride (5.19 ml, 40.7 mmol). After stirring at rt in t/n, water (15 ml) was added and stirred for 30 min. It is concentrated, then diluted with EtOAc (400 mL) and washed with saturated aqueous NaHCO3, water, 1 M HCl, water and brine. The organic layer is dried (Na2SO4) and concentrated to give the crude compound, which is used in the next step without further purification.

Step 327c. To a solution of the compound of step 327b (15.60 g, 36.6 mmol) in 2-methyl-THF (58.5 mL) was added concentrated HCl (11.70 mL, 140 mmol) at rt. The resulting clear solution was stirred at rt for 3 h. The mixture was poured portionwise into a mixture of saturated aqueous NaHCO ₃ and DCM. The aqueous layer is extracted twice with DCM. The combined organic phase is washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue is dried in vacuo to give a white solid which is used immediately in the next step.

Stage 327d. To a solution of the compound of step 327c (11.42 g, 36.6 mmol) in CH ₂ Cl ₂ (122 ml) at 0°C was added DMAP (0.447 g, 3.66 mmol), pyridine (8.87 ml, 110 mmol ) and acetic anhydride (3.79 ml, 40.2 mmol). The reaction mixture was stirred at rt overnight. Add saturated aqueous NaHCO3 (30 ml) and stir 15 min. After separation, the aqueous phase is extracted with CH2Cl2 (100 ml x 2), the combined organic phases are dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by chromatography to give the title compound (11.4 g, 93% in three steps) as an off-white solid.

Step 327e. To a solution of the compound of step 327d (4.000 g, 11.29 mmol) in acetic acid (45 ml) at rt was added manganese bis-(trifluoromethanesulfonate) (0.91 ml, 0.011 mmol) (0.0125 M in acetic acid/water 9 /1) and 2,2'-bipyridine (0.018 g, 0.113 mmol). After stirring at rt for 10 min, the peracetic acid/KOH mixture (prepared by adding 10% KOH (3.0 ml) to 35% peracetic acid (10.0 ml) was added dropwise over ~10 min, 11.79 ml, 39.5 mmol). The mixture was further stirred for 15 min at rt followed by the addition of acetone (130 ml). After 1 min at RT, the cloudy solution is filtered through a thin pad of celite and washed with acetone. The filtrate is concentrated. The crude residue was purified by chromatography (silica, EtOAc/hexanes) to give the title compound as a colorless oil (2.130 g, 51%).

Step 327f Potassium carbonate (0.799 g, 5.78 mmol). The mixture is stirred at 70° C. for 12 hours, after which it is cooled. The mixture is diluted with EtOAc and saturated NH ₄ Cl solution. The organic layer was washed with water/brine (1/1, x2), brine (x1), dried and concentrated. The crude residue was purified by chromatography (silica, EtOAc/DCM) to give the title compound (2.10 g, 70%). ESI-MS m/z=508.07, 510.07 [MH] ^- .

Stage 327g. To a solution of the compound of step 327f (40 mg, 0.078 mmol) in EtOH at 0°C was added NaBH ₄ (6 mg, 0.16 mmol). After 30 minutes, add a second portion of 2 eq. NaBH ₄ and stirred for 3 h

- 59 041670 at ct. In general ~10 eq. Add NaBH4. The mixture is concentrated to remove volatiles. The crude product was dissolved in EtOAc, washed with NH4Cl and brine, dried and concentrated to give the crude title compound (60 mg) as a white solid. The material is used in the next step without further purification. ESI-MS m/z=468.08, 470.08 [M-H]'.

Step 327h. The title compound is obtained at step 327g following the procedure described in example 130b. ESI-MS m/z=500.07, 502.07 [M-H]'.

Example 328 he

Step 328a. NaBH ₄ (360 mg, 9.6 mmol) was added to a solution of the compound of step 327f (610 mg, 1.2 mmol) in EtOH (50 ml)/Meon (15 ml) at 0°C. After 15 minutes, a second portion of NaBH ₄ (200 mg) is added. After 10 minutes, a third portion of NaBH ₄ (100 mg) was added and stirred for another 10 minutes. While still cold, the mixture is diluted by dropwise addition of HCl (0.5M) until bubbling is complete. EtOAc (200 ml) was added followed by 50 ml of water. The organic phase is washed with water and brine, dried (Na ₂ SO ₄ ) and concentrated. The crude residue was purified by chromatography (silica, EtOAc/hexanes) to give the title compound as a white solid (410 mg, 67%). ESI-MS m/z=510.09, 512.09 [MH] ^- .

Step 328b. To a solution of the compound of step 328a (382 mg, 0.77 mmol) in toluene (1 ml) was added 2(tributyl-15-phosphanylidene)acetonitrile (647 mg, 2.68 mmol) in toluene (3 ml). The mixture is heated to 95 to 100°C for 1 hour, a second portion of 2-(tributyl-15-phosphanylidene)acetonitrile (400 mg) is added and heated for another 1.5 hours. A third portion of 2-(tributyl-15phosphanylidene) is added )acetonitrile (500 mg) and heated for an additional 1.5 h, after which the mixture was cooled and concentrated; the crude product was purified by chromatography (silica, MTBE/hexanes) to give the title compound (302 mg, 80%) as a white solid. ESI-MS m/z=492.08, 494.08 [MH] ^- .

Step 328c. To the compound of step 328b (295 mg, 0.60 mmol) in acetic anhydride (5 ml) was added BF ₃ etherate (0.6 ml, 4.8 mmol) and stirred at rt for 30 min. The reaction mixture was slowly added to aqueous NaHCO ₃ (40 ml) in an ice bath. It is extracted twice with EtOAc. The combined organic phase is washed with water and brine and dried (Na ₂ SO ₄ ). After concentration, the crude residue was purified by chromatography (silica, EtOAc/hexanes) to give the title compound as a white solid (245 mg, 69%) and a mixture of bis-acetates and tri-acetates. ESIMS m/z=552.10, 554.10 [M-H] ^- (bis-acetates), m/z=594.11, 596.11[MH]-(tri-acetates).

Step 328d. To a solution of the compound of step 328c (245 mg, 0.44 mmol) in MeOH (3 mL) was added K ₂ CO ₃ (183 mg, 1.33 mmol). After 2 h add additional K ₂ CO ₃ (25 mg) and stirred for another 1 h, then diluted with EtOAc and washed with water. The aqueous phase is extracted with EtOAc. The combined organic phase is washed with brine, dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by chromatography (silica, MTBE/hexanes) to give the title compound (148 mg, 71%) as an off-white solid. ESI-MS m/z=468.08, 470.08 [MH] ^- .

Step 328e. The title compound is prepared from the compound of step 328d according to the procedure described in example 130b. ESI-MS m/z=500.07, 502.07 [M-H]-.

Example 329

To a solution of the compound of example 150 (250 mg, 0.514 mmol) in acetonitrile (5 ml) was added (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4, 5-triyl triacetate (613 mg, 1.543 mmol) and Ag ₂ CO ₃ (1.42 g, 2.57 mmol, 50 w/w% on celite). The reaction mixture is heated to 80° C. for 18 hours. The crude reaction mixture is filtered through celite, concentrated and purified by chromatography (silica, acetone/hexane) to give the title compound (195 mg, 47%). ESI-MS m/z=846.17, 848.16 (M+HCO2)-.

Example 330

To a solution of the compound of example 329 (195 mg, 0.243 mmol) in THF (3 ml), methanol (1 ml) and water (2 ml) at rt was added LiOH (50 mg, 2.09 mmol). The reaction mixture is heated to 40° C. for 1 h. The crude reaction mixture is diluted with EtOAc and acidified to pH 4 with 1 M HCl. The organic layer is washed with brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (10 mg, 6%). ESI-MS m/z=660.11, 662.11 (M-H) ^- .

Example 331

A mixture of Example 182 (159 mg, 0.30 mmol) and IBX (101 mg, 0.36 mmol) in DMSO (1.5 mL) was stirred at rt for 4 h. Na ₂ S ₂ O ₃ aqueous solution was added, and the mixture is extracted with EtOAc. The organic layer was washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC (C18 and acetonitrile/water) to give the title compound (58 mg, 36%). ESI-MS m/z=572.27, 574.27 (M+HCO2) ^- .

Example 332

Methylmagnesium bromide (0.508 ml, 1.525 mmol) is added to a solution of the compound of step 331 (230 mg, 0.436 mmol) in THF (5 ml) at -78°C. It is stirred for 3 hours at the same temperature. Add water, the mixture is extracted with EtOAc and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/acetone) to give the title compound (34.3 mg, 0.063 mmol, 15% yield) as a white solid. ESI-MS m/z=541.01, 543.01 [MH] ^- .

Example 333

Step 333a. A mixture of the compound of step 82a (112 mg, 0.25 mmol), E-hex-3-ene (630 mg, 7.5 mmol) and second generation Hoveid-Grubbs catalyst (15.7 mg, 0.025 mmol) in DCM (2 0 ml) was stirred at rt for 20 h. The mixture was purified by column chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (78 mg, 65%). ESI-MS m/z=476.13, 478.13 [MH] ^- .

Step 333b. Osmium tetroxide (0.205 ml, 2.5 % in t-butanol, 0.016 mmol) and the mixture was stirred at rt for 20 h. It was quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 3 N HCl, NaHCO ₃ and brine, dried over Na ₂ SO ₄ . filtered and concentrated to obtain a mixture of sulfone and sulfoxide, which is used without further purification.

Step 333c. m-CPBA (0.183 g, 0.85 mmol, 77%) was added to a solution of the compound of step 333b in ΝΜΠ (2 ml) and stirred at rt at 20 h. for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na2SO4, filtered, concentrated and purified by preparative HPLC (C18 and acetonitrile/water) to give the racemic product. The racemic product was separated by chiral SFC using MeOH as eluent to give the title compound (18 mg, 20%). ESI-MS m/z=542.29, 544.29 [MH] ^- .

Example 334

Step 334. Title compound (18 m/z=542.29, 544.29 [MH] ^- .

mg, 20%) isolated from example 333. ESI-MS

- 61 041670

Example 336

A solution of the compound of example 182 (1.05 g, 1.98 mmol) and IBX (0.777 g, 2.77 mmol) in DMSO (5 ml) was stirred at rt overnight. It is quenched with an aqueous solution of Na2S2O3 and an aqueous solution of NaHCO3. The reaction mixture is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The resulting crude product was purified by preparative HPLC (C18, acetonitrile/water) to give the title compound (32 mg, 3.1%) as a white solid. ESI-MS m/z=570.26, 572.26 [M+HCO2] ^- .

Example 337

Step 337a. In the solution of the compound of the stage prom. 2a (92.0 g, 676 mmol) in THF (135 ml) at 0° C. a solution of prop-1-yn-1-ylmagnesium bromide (1554 ml, 777 mmol) is added via cannula over 30 minutes. It is stirred for 5 minutes, after which the ice bath is removed. The mixture is heated to rt and kept at rt for 30 min. The reaction mixture was cooled to 0° C., saturated aqueous NH ₄ Cl (500 ml) was added followed by MTBE (500 ml). The aqueous phase is separated and extracted with MTBE (500 ml). The combined organic phases are washed with brine and dried (Na ₂ SO ₄ ). The solution was passed through a short silica gel column and concentrated to give a yellow oil (125 g, 105% yield).

Step 337b. To a solution of the compound of step 337a (50.0 g, 284 mmol) in dioxane (400 ml) and water (133 ml) at 0°C was added 2,6-lutidine (66.1 ml, 567 mmol) and sodium periodate (212 g, 993 mmol) followed by the addition of a solution of OsO ₄ (1.803 ml, 0.284 mmol, 4% in water). This mixture is heated to rt and stirred at rt for 2 days, then cooled to 0° C. and quenched with saturated aqueous Na ₂ S ₂ O ₃ . The mixture was stirred at 0°C for 1 hour, then diluted with 1 L of water and extracted with MTBE (1 L x 3). The combined organic phases are washed twice with 1 N HCl, water, aqueous NaHCO ₃ , water and brine, dried (Na ₂ SO ₄ ) and concentrated. The crude residue was purified by chromatography to give the desired product (40.2 g, 80%) as an off-white solid.

Step 337c. A solution of LAG (1 M in THF, 265 ml, 265 mmol) and DME (430 ml) is cooled to 0°C. A solution of the compound of step 337b (18.9 g, 106 mmol) in DME (80 ml) was added dropwise over 0.5 h and stirred for another 1 h. The mixture was heated and kept at 80° C. for 2 h, after which it was cooled up to 0°С. It is carefully quenched with 10 ml of water, 10 ml of 15% NaOH solution and 30 ml of water, followed by the addition of 72 g of solid Na ₂ SO ₄ and stirred for 1 h, after which it is filtered. After concentration, the crude residue was purified by chromatography to give the desired product (12.6 g, 65%) as an off-white solid.

Stage 337d. To a solution of the compound of step 337c (4.58 g, 25.1 mmol) in dichloromethane (50 ml) at rt were added pyridine (4.1 ml, 50 mmol) and benzenesulfonyl chloride (5.33 g, 30.2 mmol). The reaction mixture was stirred in t/n at rt, after which water (30 ml) was added. The mixture was stirred for 1 hour. After separation, the aqueous phase was extracted with DCM (100 ml). The combined organic phase was washed with HCl (1 M, 20 ml x2), water, NaHCO ₃ and brine and dried (Na ₂ SO ₄ ). After concentration, the crude residue was purified by chromatography to give the desired product (6.67 g, 82%) as an off-white solid.

Stage 337e. In a solution of K2OsO6-2H2O (381 mg, 1.03 mmol), (DHQ^PHAL (1.61 mg, 2.07 mmol), K2CO3 (8.58 g, 62.1 mmol) and K ₃ FeCN6 (20, 4 g, 62.1 mmol) in t-BuOH (40 ml)/water (40 ml) at 0° C. add the compound of step 337d (6.67 g, 20.69 mmol) and MeSO ₂ NH ₂ (5.90 g, 62.1 mmol).The reaction mixture is heated to rt and stirred for one day.It is cooled to 0°C and Na ₂ SO ₃ (30 g) is added. rt, then separated with EtOAc (50 ml) The aqueous phase is extracted from the extract with EtOAc The combined organic phase is washed with aqueous Na ₂ S ₂ O ₃ , water, 1 N HCl (20 ml x 2 ), water, 2 M KOH ( 10 ml x 2), water and brine x 2. It is dried (Na2SO4) and concentrated to give the desired product (7.20 g, 97%) as an off-white solid.The material is used in the next step without further purification.

Step 337f. DIPEA ( _14.11 ml, 81 mmol) and acetic anhydride ( _5.72 ml, 60 .6 mmol). The reaction mixture was stirred at rt overnight. Add saturated aqueous NaHCO ₃ (30 ml) and stir 15 min. After separation, the aqueous phase is extracted with CH2Cl2 (100 ml x2), the combined organic phases are dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by chromatography to give the title compound (8.2 g, 100% in three steps) as an off-white solid.

Stage 337g. To a clear solution of the compound of step 337f (5.09 g, 11.55 mmol) in hexafluoroisopropanol (40 ml) at rt was added manganese bis(trifluoromethanesulfonate) (20 mg ml, 0.058 mmol) and 2,2'bipyridine (0.09 g, 0.58 mmol). After stirring at rt for 10 minutes, a peracetic acid/KOH mixture (prepared by adding 10% KOH (3.0 ml) to 35% peracetic acid (10.0 ml), 11.79 was added using a syringe pump. ml, 39.5 mmol)). Add acetone (100 ml). After stirring for 10 minutes, the mixture was concentrated. The crude residue was purified by chromatography (silica, EtOAc/hexanes) to give a reduced starting material (3.1 g) and a more polar mixture (1.79 g). This more polar mixture was used in the next step without further purification.

Step 337h. Intermediate 6 was added to the highly polar mixture solution of step 337g following step 327f to obtain two batches of mixtures. One batch of the mixture contains a product such as a ketone: ESI-MS m/z=640.31, 642.31 [M+HCOO-] ^- . The other batch contains a product such as alcohol. ESIMS m/z=642.32, 644.32 [M+HCOO-] ^- .

Step 337i. The alcohol product from step 337h is worked up under the conditions of steps 328d and 328e successively and purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid. ESI-MS m/z=590.27, 592.27 [M+HCOO-] ^- .

Example 339

Step 339a. To a solution of intermediate 2a (2.29 g, 16.8 mmol) in THF (18 mL) was added ethynylmagnesium bromide (40.4 mL, 20.2 mmol, 0.5 M in THF) at 0°C and stirred at 0°C for 30 min. The reaction mixture was quenched by slow addition of aqueous NH ₄ Cl. Add MBTE and water. The mixture is separated and the organic layer is washed with water and brine. The mixture is dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to obtain the desired product (2.73 g, 100%).

Step 339b. To a solution of the compound of step 339a (1.7 g, 8.38 mmol) in THF (12 ml) at 0°C was added n-BuLi (7.38 ml, 18.4 mmol, 2.5 M in hexanes) dropwise . After 20 min at 0°C, CD ₃ I (0.69 ml, 10.9 mmol) was added and the mixture was stirred at rt for 20 h. The reaction mixture was quenched with slow addition of aqueous NH ₄ Cl. Add MBTE and water. The mixture is separated and the organic layer is washed with water and brine. The mixture is dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the desired product (1.09 g, 65%).

Step 339c. To a solution of the compound of step 339c (0.986 g, 5.5 mmol) in dioxane-water (12/4 ml) was added 2,6-lutidine (1.45 ml, 11 mmol) and OsO ₄ (0.18 ml, 2. 5% solution in t-butanol, 0.017 mmol). The mixture is cooled and NaIO ₄ (4.12 g, 19.25 mmol) is added. The suspension is stirred at rt for 16 h. An aqueous solution of Na2S2O3 is added. The mixture is stirred for 1 hour and filtered through celite. The mixture is extracted with MBT/hexanes. The organic phase was washed with water, 1 N HCl, saturated NaHCO ₃ and brine, dried over Na 2 SO 4 , filtered and concentrated to give the crude product (0.64 g, 64.2%).

Stage 339d. MeONa (23 mg, 0.42 mmol) was added to a solution of the compound of step 339c (0.64 g, 3.53 mmol) in CD3OD (10 mL) and the mixture was stirred at rt for 3 h. dissolved in CD ₃ OD (5 ml) and stirred for 3 h. The same reaction cycle was repeated two more times and the solution was quenched with D2O. The mixture is extracted with MBTE. The organic layer was washed with D ₂ O and saturated NaCl in D ₂ O, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound (0.62 g, 95%).

Stage 339e. To a solution of LiBH ₄ (3.35 ml, 6.70 mmol, 2 M solution in THF) at -40° C. was added the compound of step 339d (0.62 g, 3.35 mmol) in MBTE (20 ml). The resulting solution is heated to 0°C for 1 h, the Reaction mixture is quenched by slowly adding aqueous NH ₄ Cl. Add MBTE and water. The mixture is separated and the organic layer is washed with brine. The mixture is dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the title compound as a white solid (0.45 g, 72%).

Stage 339f. A solution of the compound of step 339e (0.45 g, 2.40 mmol), LAH (6.01 ml, 6.01 mmol, 1M in THF) and MeONa (26 mg, 0.48 mmol) in DME (12 ml) was heated and incubated at 80°C for 2 hours. The reaction mixture is cooled to 0°C, carefully quenched with 0.23 ml of water and 0.23 ml of 15% NaOH solution, incubated for 10 min, then add 0.69 ml water, followed by the addition of 5 g of solid Na2SO4, and stirred for 1 h, after which it was filtered. The filtrate is concentrated. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the title compound (0.32 g, 70%).

Stage 339g. Pyridine (0.33 ml, 4.3 mmol) and 4-toluenesulfonyl chloride (0.387 g, 2.02 mmol). The reaction mixture is stirred at rt in t/n, and water is added. The mixture is stirred for 1 hour. After separation, the aqueous phase is extracted with DCM. The combined organic phase is washed with 1 N HCl, water, NaHCO ₃ and brine, dried over Na ₂ SO ₄ and concentrated. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the title compound (0.52 g, 90%).

Step 339h. Into a solution of ^OsO^O (27 mg, 0.073 mmol), (DHQ^PHAL (85 mg, 0.109 mmol), K2CO3 (0.604 g, 4.37 mmol) and K ₃ FeCN ₆ (1.44 g, 4.37 mmol) in t-BuOH (7.0 ml)/water (40 ml) at 0° C. add the compound of step 339g (0.50 g, 1.46 mmol) and MeSO ₂ NH ₂ (0.277 g, 2.91 mmol The reaction mixture was allowed to warm slowly to rt and stirred for 3 days.The mixture was cooled to 0°C followed by the addition of Na ₂ SO _3. The mixture was stirred 15 min at 0°C, then 1 h at rt, after which it was separated with EtOAc The aqueous phase is extracted from the extract with EtOAc The combined organic phase is washed with aqueous Na ₂ S ₂ O ₃ , water, 1 N HCl, water, 2 M KOH, water and brine, dried (Na ₂ SO ₄ ), filtered and concentrated The crude product was purified by chromatography (silica, hexanes/EtOAc), then dispersed in MeOH (5.6 ml) for 16 h and filtered to collect the desired product as a white solid (0.39 g, 70%) .

Stage 339i. A solution of the compound of step 339 h (0.39 g, 1.03 mmol), intermediate 6 (0.316 g, 1.06 mmol) and K2CO3 (0.143 g, 1.03 mmol) in DMF (1.1 ml) was stirred at 70°C for 16 hours It is diluted with EtOAc, and the mixture is washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (0.37 g, 71%). ESI-MS m/z=503.16, 505.16 [MH] ^- .

Stage 339j. A solution of the compound of step 339i (0.37 g, 0.73 mmol) and m-CPBA (0.575 g, 2.56 mmol, 77%) in ΝΜΠ (3.0 mL) was stirred at rt for 24 h. Aqueous Na was added ₂ S ₂ O ₃ , NaHCO ₃ and a few drops of Et ₃ N and stirred at rt for 1 h. It is extracted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ , filtered and recrystallized from MeOH to give the title compound (0.33 g, 84%). ESI-MS m/z=535.15, 537.15 [MH] ^- .

Example 340

Step 340a. A solution of the compound of step 337c (3.2 g, 17.6 mmol) and IBX (6.8 g, 24.3 mmol) in DMSO (20 ml) was stirred at 45°C for 14 h. ₂ S ₂ O ₃ and the mixture is extracted with EtOAc. The organic layer is washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the desired product (2.3 g, 72%).

Step 340b. MeONa (138 mg, 2.55 mmol) was added to a solution of the compound of step 340a (2.3 g, 12.8 mmol) in CD3OD (20 ml) and the mixture was stirred at rt for 3 h. dissolved in CD3OD (20 ml) and stirred for 3 hours. The same reaction cycle was repeated two more times and the solution was quenched with D2O. The mixture is extracted with MBTE. The organic layer was washed with D2O and saturated NaCl in D2O, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound (2.0 g, 85%).

Step 340c. To a solution of LiBH ₄ (10.8 ml, 21.6 mmol, 2 M solution in THF) at -40° C. was added the compound of step 340b (2.0 g, 10.8 mmol) in MBTE (60 ml). The resulting solution is heated to 0° C. for 1 h. The reaction mixture is quenched by slowly adding aqueous NH ₄ Cl, then diluted with MBTE and water. The mixture is separated and the organic layer is washed with brine. The mixture is dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the title compound as a white solid (1.4 g, 69%).

Step 340d. Pyridine (0.28 ml, 3.48 mmol) and 3-nitrobenzenesulfonyl chloride (0.385 g, 1.74 mmol) are added to a solution of the compound of step 340c (0.216 g, 1.16 mmol) in dichloromethane (1.5 ml) at rt. . The reaction mixture is stirred at rt in t/n. Water is added, then stirred for 1 hour. After separation, the aqueous phase is extracted with DCM. The combined organic phase is washed with 1N HCl, water, NaHCO3 and brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/MBTE) to give the title compound (0.37 g, 86%).

Step 340e. Solution of the compound of step 340d (0.36 g, 0.97 mmol), methyl 4-chloro-3-mercaptobenzoate (0.206 g, 1.02 mmol) and Cs ₂ CO ₃ (0.316 g, 0.97 mmol) in DMF (1.1 ml) stirred at 80° C. for 16 h. It was diluted with EtOAc and the mixture was washed with water and brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to semi

- 64 041670 the title compound as a white solid (0.25 g, 69%).

Step 340f. To a solution of the compound of step 340e (0.25 g, 0.674 mmol) in THF (3.0 ml) was added LiOH (2.7 ml, 1.35 mmol, 0.5 M solution in water) and the mixture was stirred at rt at within 3 hours the Reaction mixture is quenched by adding 1 N HCl. Add EtOAc and water. The mixture is separated and the organic layer is washed with brine. The mixture was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound as a white solid (0.23 g, 96%). ESI-MS m/z=355.24, 357.24 [MH] ^- .

Stage 340g. To a solution of the compound of step 340f (0.23 g, 0.644 mmol), 3,4-difluorobenzene-2,6-D2-amine (0.127 g, 0.967 mmol), DMAP (0.039 g, 0.322 mmol) in DMF (2.5 ml) add EDC (0.185 g, 0.967 mmol) and stir at rt for 16 h. It is diluted with EtOAc and the mixture is washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound (0.26 g, 86%). ESI-MS m/z=468.31, 470.31 [MH] ^- .

Step 340h. Osmium tetroxide (0.28 ml, 0.055 mmol, 5% in water), and the mixture is stirred at rt for 20 h. It is quenched with aqueous Na ₂ SO ₃ , extracted with EtOAc, washed with water, 1 N HCl, NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered and concentrated to obtain a mixture of sulfone and sulfoxide, which is used without further purification.

Step 340i. To a solution of the compound from step 340h in ΝΜΠ (2.5 mL) was added m-CPBA (0.41 g, 1.83 mmol, 77%) and stirred at rt for 20 h. Add aqueous Na2S2O3, NaHCO3 and a few drops of Et3N and stirred at rt for 1 h. The mixture was extracted with EtOAc, washed with water and brine, dried over Na2SO4, filtered and concentrated to give the title compound (0.27 g, 96%). ESI-MS m/z=580.33, 582.33 (M+HCO2)'.

Step 340j. A mixture of the compound of step 340i (362 mg, 0.675 mmol) and IBX (227 mg, 0.81 mmol) in DMSO (5.0 mL) was stirred at rt for 4 h. Aqueous Na2S2O3 was added and the mixture was extracted with EtOAc. The organic layer was washed with water and brine, dried over Na ₂ SO ₄ , filtered, concentrated and purified by preparative HPLC (C18 column, acetonitrile/water) to give the title compound (150 mg, 41%). ESI-MS m/z=578.32, 580.32 (M+HCO2)'.

Example 341 he he

Step 341a. 1-Methyl-1H-pyrrole-2-carboxylic acid (3 g, 23.98 mmol) was dissolved portionwise in chlorosulfonic acid (12 ml, 179 mmol) in an ice bath. The reaction mixture is stirred for 70 minutes at the same temperature. The mixture was slowly quenched with ice/water (100 ml) (very reactive!!) and stirred for 15 minutes. The precipitate is filtered, rinsed with water and the resulting solid is dissolved in EtOAc and dried over NaSO4. The solvent was removed in vacuo and dried overnight in vacuo to give the title compound (3.60 g, 16.10 mmol, 67% yield) as an off-white solid.

Step 341b. To a mixture of the compound of step 341a (1 g, 4.47 mmol) in DCM (20 ml) and DMF (0.02 ml) was added oxalyl chloride (11.18 ml, 22.36 mmol). The reaction mixture was stirred at rt for 5 h. The solvent was removed in vacuo, then diluted with benzene (x3) to give the title compound (1.08 g, 100% yield) as a pale yellow solid.

Step 341c. To a solution of the compound of step 341b (400 mg, 1.652 mmol) in toluene (15 ml) at rt was added 3,4-difluoroaniline (0.164 ml, 1.652 mmol) in toluene (1 ml). The mixture is heated to 110° C. and stirred for 1 h, then cooled to rt and stirred for 16 h. The solid is filtered off and the resulting filtrate is removed in vacuo, and the solid is used without further purification (553 mg, 100% yield). ).

Step 341d. To a suspension of the compound of step 341c (550 mg, 1.643 mmol) in toluene (10 mL) was added triphenylphosphine (2.16 g, 8.22 mmol). The reaction mixture is heated to 85° C. and stirred for 6 hours. The reaction mixture is cooled to rt and 10 ml of H2O are added. The organic layer was washed twice with water (5 ml) followed by washing with 1N NaOH. The aqueous layer is collected. The aqueous layer is acidified with 1 N HCl to pH 4. After addition of HCl, it turns into a white emulsion. It is extracted with EtOAc and the resulting mixture turns into a clear colorless solution. The organic layer was collected and dried over Na2SO4 to give the title compound (220 mg, 0.820 mmol, 50% yield) as a white solid.

Step 341e. Cesium carbonate (197 mg, 0.606 mmol) was added to a solution of the compound of step 341d (130 mg, 0.485 mmol) and the compound of step 337d (187 mg, 0.505 mmol) in DMF (0.5 ml). The reaction mixture is heated to 70°C and stirred for 17 hours. The mixture is extracted with EtOAc, dried over Na2SO4, and then purified by column chromatography (silica, hexanes/EtOAc) to obtain the target

- 65 041670 units (170 mg, 0.364 mmol, 72% yield) as a white solid.

Step 341f. m-CPBA (50.4 mg, 0.225 mmol, 77%) is added to a solution of the compound of step 341e (35 mg, 0.075 mmol) in ΝΜΠ (0.3 ml) at 0°C. The reaction mixture is slowly heated to rt and stirred for 15 h. Saturated Na ₂ S ₂ O ₃ solution and NaHCO ₃ solution are added and the resulting mixture is stirred at rt for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O solution ₃ , NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes/EtOAc) to give the title compound (24.0 mg, 0.048 mmol, 64% yield) as a white solid. ESI-MS m/z=497.52, 499.53 [MH]'.

Example 344 about

_F I n B I °

Step 344a. To a solution of the compound of step 307a (500 mg, 0.71 mmol) in 2-methyltetrahydrofuran (3 mL) was added (R)-1-aminopropan-2-ol (160 mg, 2.12 mmol), followed by stirring at 75°C. within one day. The crude product is diluted with EtOAc, washed twice with water, washed twice with brine, dried (Na ₂ SO ₄ ) and concentrated. The crude product was purified by chromatography (silica, MeOH/DCM) to give the title compound (255 mg) as a mixture of the title compound with other impurities. ESI-MS m/z=479.27, 48127 [MH]'.

Step 344b. To a solution of Step 344a (255 mg, 0.53 mmol) in 2-methyltetrahydrofuran (2 ml) was added CDI (112 mg, 0.69 mmol) and TEA (0.15 ml, 1.06 mmol). The reaction mixture is heated to 50°C for 1 h, then cooled and concentrated. The crude product is purified by chromatography (silica, EtOAc/hexanes) to give the title compound as a mixture with other impurities. ESI-MS m/z=505.26, 507.26 [M-H]'.

Step 344c. The compound of step 344b is worked up under the conditions described in step 130b and purified by preparative HPLC (C-18, acetonitrile/water) to give the title compound as a white solid. ESI-MS m/z=537.26, 539.26 [M-H]'.

Example 345

Step 345a. Triethylamine (0.586 ml, 4.20 mmol) and Ac ₂ O (0.238 ml, 2.52 mmol) were added to a solution of the compound of step 182 (1.046 g, 2.100 mmol) in CH2C12 (21 ml) at rt, then the mixture was kept at rt during the night. Then the reaction mixture is divided between an aqueous solution of NaHCO ₃ and CH2C12. The combined organic phases are dried over Na2SO4 and concentrated, and the crude product is purified by flash column chromatography to give a mixture (415 mg) of two inseparable monoacetylated products. ESI-MS m/z=584.13, 586.13 [M+HCO2] ^- .

Step 345b. To a solution of the compound of step 345a (415 mg, 0.768 mmol) in CH2C12 was added 2-iodoxybenzoic acid (430 mg, 1.54 mmol). The mixture is kept at rt overnight, then a saturated aqueous solution of Na ₂ S ₂ O ₃ is added. The mixture is partitioned between EtOAc and water. The combined organic phases are washed with brine, dried over Na ₂ SO ₄ and concentrated. The resulting crude product was purified by chromatography (silica, hexanes/EtOAc) to give the title compound as a white solid (39 mg, 8.2% in 2 steps). ESI-MS m/z=582.12, 584.12 [M+HCO2] ^- .

Step 345c. K2CO3 (10 mg, 0.073 mmol) was added to a solution of the compound of step 345b (39.1 mg, 0.073 mmol) in THF (0.3 ml) and MeOH (1.1 ml) at rt. The mixture is then kept at rt for 1 h. The reaction mixture is diluted with MTBE, washed with water, dried over Na ₂ SO ₄ and concentrated to give the crude title product (19 mg, 53%) as a white solid, which is used in the next step. without additional cleaning. ESI-MS m/z=540.11, 542.11 [M+HCO2]'.

Stage 345d. To a solution of the compound of step 345c (19 mg, 0.038 mmol) in CH2C12 (1.9 ml) at rt was added m-CPBA (34.3 mg, 0.153 mmol, 77%) and stirred at rt overnight. The reaction mixture was quenched with saturated aqueous Na2S2O3 and aqueous NaHCO3. The reaction mixture is partitioned between EtOAc and the aqueous phase. The combined organic phases are washed with brine, dried over Na2SO4 and concentrated. The resulting crude product was purified by chromatography to give the title compound as a white solid (11 mg, 29% in 2 steps). ESI-MS m/z=572.09, 574.09 [M+HCO2] ^- .

- 66 041670

Example 346 he he

Step 346a. To a solution of ethynylcyclopropane (1.864 ml, 22.03 mmol) in THF (4.50 ml) at -78°C was added n-BuLi (8.81 ml, 22.03 mmol, 2.5 M in THF). The resulting mixture was stirred for 1 hour at the same temperature. In the reaction mixture at -78°C add a solution of the compounds of stage prom. 2a (2.0 g, 14.68 mmol) in THF (9 ml). The light yellow solution obtained is heated slowly to rt and stirred for 2 h. Saturated NH ₄ Cl solution is added. The mixture was extracted with MTBE and dried over Na ₂ SO ₄ to give 2.95 g (99% yield) of the title compound as a yellow oil, which was used without further purification.

Step 346b. To a solution of the compound of step 346a (2.97 g, 14.68 mmol) in dioxane (122 ml) and water (25 ml) at 5°C was added 2,6-lutidine (3.42 ml, 29.4 mmol) and sodium periodate (10.99 g, 51.4 mmol). OsO ₄ (0.921 ml, 0.073 mmol, 2.5% in t-BuOH) was added and the resulting mixture was stirred at rt for 15 h. Na ₂ S ₂ O ₃ (50 ml) and EtOAc (100 ml) were added. The mixture is stirred for 30 minutes, then filtered. The solid is washed with EtOAc (x3). The filtrate is collected and washed with 0.5 N HCl (x4). The aqueous layer is extracted from the extract with EtOAc and the combined organic layers are dried over Na ₂ SO ₄ . The crude material was purified by chromatography (silica, hexanes/acetone) to give the title compound (2.25 g, 11.01 mmol, 75% yield) as a light yellow oil.

Step 346c. To a solution of LiBH ₄ (13.77 ml, 27.5 mmol, 2 M in THF) in MTBE (22.03 ml) at -50°C was added a solution of the compound of step 346b (2.25 g, 11.01 mmol) in MTBE (5 ml). The reaction mixture was stirred at -50°C for 3 hours Add NH ₄ Cl (50 ml). The mixture was extracted with EtOAc (100 ml), washed with brine, dried over Na ₂ SO ₄ and purified by chromatography (silica, hexanes/EtOAc) to give the title compound (1.62 g, 7.85 mmol, 71% yield) as a solid white substances.

Step 346d. To a mixture of LAH (13.77 ml, 13.77 mmol, 1 M in THF) and NaOMe (0.074 g, 1.377 mmol) in DME (15 ml) was added at rt a solution of the compound of step 346c (1.42 g, 6.88 mmol) in DME (2 ml). The reaction mixture is heated to 80° C. and stirred for 3 hours. The reaction mixture is cooled to 0° C., carefully quenched with water and 1 N NaOH solution. The mixture is filtered. The filtrate is extracted with MTBE and dried over Na ₂ SO ₄ . The resulting material is recrystallized with MTBE and hexane to give the title compound (1.30 g, 6.24 mmol, 91% yield) as a white solid.

Step 346e. To a solution of the compound of step 346d (1.6 g, 7.68 mmol) in DCM (10 mL) at 0°C was added pyridine (1.243 mL, 15.36 mmol) and 4-nitrobenzenesulfonyl chloride (1.904 g, 9.99 mmol) . The reaction mixture is slowly heated to rt and stirred for 16 h. Water is added, the mixture is extracted with DCM and dried over Na ₂ SO ₄ . The crude material was purified by column chromatography (silica, hexanes, EtOAc) to give the title compound (2.10 g, 5.79 mmol, 75% yield) as a colorless oily liquid.

Step 346f. To a mixture of t-BuOH (29.0 ml) and water (29.0 ml) at 0°C was added potassium osmate dihydrate (0.064 g, 0.174 mmol), (DHQ) ₂ PHAL (0.203 g, 0.261 mmol), hexacyanoferrate( III) potassium (5.72 g, 17.38 mmol) and potassium carbonate (2.402 g, 17.38 mmol) followed by the addition of methanesulfonamide (3.31 g, 34.8 mmol) and the compound of step 346e (2.1 g , 5.79 mmol). The reaction mixture was slowly heated to rt and stirred for 50 h. Na ₂ S ₂ O ₃ (3.7 g) was added at 0° C. and the resulting mixture was stirred at rt for 1 h. EtOAc was added, the solid was filtered through Celite and washed with EtOAc (x2). The organic layer was extracted with EtOAc, washed with 1 N HCl (x3), followed by 2 N K2CO3 and brine, and dried over Na ₂ SO ₄ to give 1.85 g (4.67 mmol, 81% yield) of the desired product as a solid. whitish substance, which is used without further purification.

Stage 346g. To a solution of the compound of step 346f (1.85 g, 4.67 mmol) in DMF (15.55 ml) was added K2CO3 (0.632 g, 4.57 mmol) at rt. 4-Chloro-N-(3,4-difluorophenyl)-3-mercaptobenzamide (1.468 g, 4.90 mmol) is added. The resulting mixture is heated to 75° C. and stirred for 15 hours. The reaction mixture is cooled to rt, water is added and extracted with EtOAc (x3), washed with brine and dried over Na ₂ SO ₄ . The crude material was purified by chromatography (silica, hexanes/EtOAc) to give the title compound (1.91 g, 3.64 mmol, 75% yield) as an off-white solid.

Step 346h. To a solution of the compound of step 346g (1.91 g, 3.64 mmol) in ΝΜΠ (10 ml) at 0°C was added m-CPBA (2.451 g, 10.93 mmol, 77%). The mixture is heated to rt and stirred for 15 h. The reaction mixture is quenched with saturated Na2S2O3 solution and NaHCO3 solution and stirred for 1 h. The mixture is extracted with EtOAc, washed with saturated Na ₂ S ₂ O ₃ solution, NaHCO ₃ solution (x2) and brine, and dried over Na ₂ SO ₄ . The crude material was purified by chromatography (silica, hexanes/acetone) to give the title compound (1.26 g, 2.27 mmol, 42% yield) as a white solid. ESI-MS m/z=555.02, 557.02 [MH] ^- .

- 67 041670

Example 347

Step 347a. n-BuLi (8.81 ml, 22.03 mmol, 2.5 M in THF). The resulting mixture was stirred for 1 hour at the same temperature. In the reaction mixture at -78°C add a solution of the compounds of stage prom. 2a (2.0 g, 14.68 mmol) in THF (10 ml). The light yellow solution obtained is heated slowly to rt and stirred for 2 h. Saturated NH ₄ Cl solution is added. The mixture was extracted with MTBE and dried over Na ₂ SO ₄ to give 3.0 g (99% yield) of the title compound as a yellow oil, which was used without further purification.

Step 347b. To a solution of the compound of step 347a (3.0 g, 14.68 mmol) in dioxane (122 ml) and water (25 ml) at 5°C was added 2,6-lutidine (3.42 ml, 29.4 mmol) and sodium periodate (10.99 g, 51.4 mmol). Osmium tetroxide (0.921 ml, 0.073 mmol, 2.5% in tBuOH) was added and the resulting mixture was stirred at rt for 16 h. Na ₂ S ₂ O ₃ (50 ml) and EtOAc (100 ml) were added. The mixture is stirred for 30 minutes, then filtered. The solid is washed with EtOAc (x3). The filtrate is collected and washed with 0.5 N HCl (x4). The aqueous layer is extracted from the extract with EtOAc and the combined organic layer is dried over Na ₂ SO ₄ . The crude material was purified by chromatography (silica, hexanes/acetone) to give the title compound (1.60 g, 7.76 mmol, 53% yield) as a light yellow oil.

Step 347c. To a solution of LiBH ₄ (9.70 ml, 19.39 mmol, 2 M in THF) in MTBE (15.51 ml) at -50°C was added a solution of the compound of step 347b (1.60 g, 7.76 mmol) in MTBE (5 ml). The reaction mixture was stirred at -50°C for 3 hours Add NH ₄ Cl (50 ml). The mixture is extracted with EtOAc (100 ml), washed with brine and dried over Na ₂ SO ₄ . The crude material was purified by chromatography (silica, hexanes/EtOAc) to give the title compound (1.16 g, 5.58 mmol, 72% yield) as a white foamy solid.

Stage 347d. To a mixture of LAH (11.16 ml, 11.16 mmol, 1 M in THF) and sodium methoxide (0.060 g, 1.116 mmol) in DME (15 ml) was added at rt a solution of the compound of step 347c (1.162 g, 5.58 mmol ) in DME (2 ml). The reaction mixture is heated to 80° C. and stirred for 3 hours. The reaction mixture is cooled to 0° C., carefully quenched with water and 1 N NaOH solution. The mixture is filtered. The filtrate is extracted with MTBE and dried over Na ₂ SO ₄ . The resulting material is recrystallized with MTBE and hexane to give the title compound (1.06 g, 5.02 mmol, 90% yield) as a white solid.

Stage 347e. The title compound was obtained from the compound of step 347d by the procedure described in Example 346e as a cloudy oil (1.51 g, 4.14 mmol, 87% yield).

Stage 347f. The title compound was obtained from the compound of step 347e by the procedure described in Example 346f as an off-white solid (1.01 g, 2.53 mmol, 61% yield).

Stage 347g. The title compound was obtained from the compound of step 347f by the procedure described in Example 346g as an off-white solid (0.87 g, 1.65 mmol, 65% yield).

Step 347h. The title compound was obtained from the compound of step 347g by the procedure described in Example 346h as a white solid (0.74 g, 1.32 mmol, 80% yield). ESI-MS m/z=557.03, 559.02 [MH] ^- .

Example 348 he he

Step 348a. To a solution of the compound of Example 82a (50 mg, 0.111 mmol) in DCE (2 mL) at rt was added tert-butyl 2-oxo-5-vinyloxazolidine-3-carboxylate (71.1 mg, 0.333 mmol) and a second generation Hoveid Grubbs catalyst. (6.96 mg, 0.011 mmol). The reaction mixture is degassed by passing N2 for 5 min, heated to 60° C. and stirred for 15 h. The reaction mixture is cooled to rt. Add water. The mixture is extracted with DCM and dried over Na2SO4. The crude material was purified by chromatography (silica, hexanes/EtOAc) to give the title compound (45 mg, 0.071 mmol, 64% yield) as an off-white solid.

Step 348b. To a solution of the compound of step 348a (45 mg, 0.071 mmol) in acetone (3 ml) and water (0.8 ml) was added NMO (33.2 mg, 0.283 mmol) and OsO ₄ (178 μl, 0.014 mmol, 2.5 % in t-BuOH). The reaction mixture was stirred for 16 hours. It was quenched with saturated Na ₂ S ₂ O ₃ solution, extracted with EtOAc, washed with brine and dried over Na ₂ SO ₄ . The crude material is used without further purification.

Step 348c. To a solution of the compound of step 348b (50 mg, 0.071 mmol) in DCM (1 mL) was added TFA (0.027 mg, 0.355 mmol) at rt. The reaction mixture is stirred for 2 hours. The solvent is removed.

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The mixture is extracted with DCM, washed with saturated NaHCO3 solution and dried over Na2SO4. The crude material was purified by column chromatography (silica, MeOH/DCM) to give the title compound (2:1 mixture of isomers, 14.5 mg, 0.024 mmol, 34% yield) as an off-white solid. ESI-MS m/z=600.02, 602.02 [MH] ^- .

The following examples were prepared using procedures similar to those described above.

Example Structure ESIMS (MH)' or (M+H) ⁺ 2 L % th 587.12.589.12 8 12 13 15 16 17 22 23 he F ί” N GL Τι TS ° _F βΎ A ⁰ but ^f ϊ ^f yS ° ' ⁰ A ^s=o N _c OH ,. o—\ Ll ° η H 1 ° , ^0H FJ ^H Η ΓΛ YAA^oCh ° - il TC ⁰ OH ^4--0 JCL u_j=oI ⁰ “° ^F H . /V' y° h [1 1 ⁰ a, 672.13.674.13 569.09, 571.09 573.09 _, 575.09 599.123.601.121 714.19.716.19 714.19.716.18 641.13, 643.13 15 (M+HCO2) 24 pH F ? ^H H frt rt o rt^ ^N Y° ^0H JL Th a ⁼ o 'I ° h U 1 ° 675.13, 677.13 (M+HCO2)

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25 but ,5 Vn ° ru ^b JI JL sso ' F c || sc racemic 603.10, 605.10 thirty Rr ^OH A ^and Q'° 532.05 31 rr ^0H CF ₃ j Ά - ^and 444.08, 446.06, 537.20 [Μ+ΝΗ ₄ ] ⁺ 39 '0 ' IZ c o ,sh—L - O^ll Vs Λ 0 605.07, 607.07 40 IZ c o _χ ω - £ - o*n Vs % 4 At 639.08, 641.08 41 _F G G> I _____J... ^NH. J<Z Ϊΐ u ГЦ 8 η Η 1 ⁰ 641.13, 643.13 (M+HC0 ₂ )" 43 e 2 z g ABOUT- /< o bH ιι<ο _ $ ZX >~c^ \L 494.20 44 S Z Cha i<° f S 466.20

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45 z about i ιζ g - 0*11 VS o >/ S—o z 3 in 494.20 [M + NH ₄ ] ⁺ 46 A CL u<° Ν..ι(η ^ζ O i o 486.15 [M+H] ⁺ , 503.15 [M+NH ₄ ] ⁺ 47 V 2 Z? O- /( O <A lUO o £2 i 450.20 48 & IZ A o ,ω>··^ - 0*11 VS A = \ Z Z 3..... -sA 630.12, 632.12 49 o— X ° H H- x2 % A A i. And. 646.11, 648.11 50 he _n Ά o Xda^z^l ^{0 n 1} n [1 I ° 658.16, 660.16 53 it A n [1 1 ° 584.8, 586.8 54 he o M and ⁿ ' n U 1 ° A^CI 586.8, 588.8

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55 OH O I A n U 1 ⁰ 586.8, 588.8 57 he o he XI A^DL ° ^{n F} ” ΐΓΧ'° 616.8,618.8 58 he o _F i XX JL-jiN ⁿ n u 1 ° 568.8, 570.8 59 bl ΑΑβ ρΑΑ/γ^Α» ⁴¹ ' ° * ^® 675.13, 677.13 (M+HCO ₂ ) 60 OH OH arcHA 4^1^° racemic 606.11.608.11 62 F Г η F 1 Aw'' ^/O ------ xx n Π 1 ° ^s^ci 546.08, 548.08 63 X oh ^! ΐ b. X y IL IL 576.09, 578.09 65 he o R X. ^bry'''*'^ ⁴ N^S^ ^X OH P 8 ^H ™ _ρ ΛΑ _ν ^ί η Π 1 ° 602.8, 604.8

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66 d IZ X - ο^ιι v> Λ \ Z Z y 627.11, 629.11 (M+HCO2)' 67 X X 2 \ ABOUT- Wu zl 661.12, 663.12 (M+HCO2)' 68 he he F^l η Xd ^he U ^JL _t 'Relative stereochemistry 650.11, 652.11 (M+HCO2)' 69 D o ^aVyb^ 484.20 [M+H] ⁺ , 501.20 [M+NH ₄ ] ⁺ 71 he o ₽A XX dL ^{n x f} id ⁰ 600.8, 602.8 72 A ΖΖ X O _ζ Μι κ - ο^ιι vs A s=° zz ъ 598.8, 600.8 73 OH O g—Λ A gA ^L ΝΎν Η Π 1 ° 598.8, 600.8 75 F ___l° ^H Η Λ Ά1 ^H ^s ~ 712.18.714.18

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77 U' IZ O xCOm-s. “O ^X N VS O ) / O \ zz 760.20, 762.20 [M+NSOGG 79 H.O. r -J s b Ά » pdf' t\ ^Η IL 1 ° 718.16, 720.16 [M+HCO2G 80 OH Η Η F III F Α ' ^,χΝ >ι^^ ^Ν >< ^0Μβ λΧ ° ° η || 1 ο 676.14, 678.14 (M+HCO2) 81 G ζ-ζ ζ) To $ N LL U. 676.14, 678.14 (M+HCO2) 83 & ζζ A % \ ζζο _% °5„.ъ ζζ g 673.13.675.13 84 Ο^ΝΗ ₂ OH _u \ 0 ί Ν A L AvrF ^Hn 687.15.689.15 85 ζζ A ο xW”(L Ά 4 * Z \ ZZ Hh Ύ Ο 614.09, 616.09 86 He ,, . ο 1 Ν Λ Α ^f A i 5 η ^ο JL ^s=o Ν n U JL ° 630.13.632.13

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87 g zz \ b RR IUO _ £ zz l 598.8, 600.8 88 Q )=ο χζ ъ BH IUO_ £ LL LL 595.8, 597.8 89 g XZ \ X ? ABOUT- 11° BH 1b0 _ £ A' 612.8,614.8 90 F? ^H Η o A^cG ” 612.8,614.8 91 \ /° XZ\o— /( ABOUT BH IUO_ Z t A 612.8,614.8 92 ZZ O _ζ <Λ κ - o*n UCH A. I Ζ 584.7, 586.7 93 ZZ ² o*n Vs A. U° O X 626.8, 628.8 94 OH o ^F J II Ύί ίϊ JI JL ____. ^s=o N vo η H 1 ^{0 4} ---^Cl 610.8,612.8 95 zz ² o*rfs \ zz “r° o 614.09, 616.09

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96 F? ^H ? ^Η Η sA, ο 43/ Υ DD I ^{0 0} w n η 'X ⁿ Y 1 ° ^^Cl 646.12, 648.12 97 'Λ ZZ - O II vs l ,** Z \ ZZ Ύ o 614.10, 616.10 98 IZ 0 x "P - 0^11 VS d. Z \ o o g No. 560.11, 562.11 99 OH FI _o chl. xDh Jkx^^xS.=O F ^-^ N Χ^Ί< A and π ι ° 604.15, 606.15 100 Π IZ r O O=SL"^ ^S—o Z g f 548.00 101 o ^ MJ \u003d ° y # O xSP * ^ - 0 * 11 VS Lo k ^x o 546.15 102 Mj=° θ < ^w,, 4s h. E ¹ CD 547.20 103 Ο-ΊΙ V> 0 >/ ^A S— ⁰ s ¹ φ 526.15

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104 OME? ^{H H} 1! 1 482.15 105 IZ X Ο _X "'" S - oll V) O iJ S - o ; Z s F 487.15 [M + NH ₄ ] ⁺ 107 OH ο °γ-°χ 'b '° ^h %^C1 702.15.704.15 BY xz tf O /SL "K - Ο-ΊΙ V> 4.$=0 IZ 584.8, 586.8 111 he o ^0" Υί u ^H h U 1 ° 598.8, 600.8 112 it l h 0 1° 586.8, 588.8 113 xz Οχ<Λ···Κ. - ο-Ίΐ Cs A. $=ό zz about z 586.8, 588.8 114 he o %L ZirAA»^ ⁰ * XI Η ϋ 1 ° 586.8, 588.8 115 '0 ' zz X ° a ⁰ ” Vs - O^ll VS X $=° zz s Φ 600.8, 602.8

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116 zz / Ο - 0^11 Vs 4. % zz about s ® 612.8,614.8 117 & zz / ABOUT - OL1 vs 4. zz o z 598.8, 600.8 118 OH 0 F 1 II 1 I /-___J... JI x4 .OMe A. /4/ '^ N Il Ί ° :Cl ^H X<^s N FN Sff x\ hh ι ° 600.8, 602.8 119 he o F 1 II 1 G-----1-.. Ek ,OMe R A. /2Ty '^ N S<S< Ss o ^GT ^{n 11} II : H 0 xk^A x*k^S^xS =OX ΐ— Ν STλ' η Η I ° 600.8, 602.8 122 OH HO f I Y1 u ^*4 F-^NArW ^{N H} H 1 ⁰ 590.12, 592.12 123 F ε o \ o— /( O FA iuo _ ^*·>|<0 ^x o l l 516.08.518.08 126 OH Ac OH F ι ι : 1 Г------- >4. F^ A. n, ZZTy ' ^x 's^'S. Y1 S = N _f A>s _n AfvV 4 h U 1 ° ^4^01 601,13,603,13 127 zz y - O^ll VS A \ ZI "5? O 612.11.614.11 128 o \ ^zx ZZ ^s ir- Ga n=o _ 7···«' and A IL IL 658.12, 660.12 [M- НСО2Г

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129 he is about ηΜ^ ^ν^υ ^οη 601,13,603,12 131 OH HO I ^p D n Η 1 ° S^'ci 560.11.562.11 132 m>° o _χ ω···v. - 0^11 VS <o 3 ¹ Φ 673.0, 675.0, 547.05 133 F ____° ^H ° RF ' ^Me C IL JL ______s=oN h jpY'° 4^N 547.20 134 IZ & - 6Hi VS <os ¹ F 468.15 135 01 s - o ^x n Vs C-Ο 3 ¹ 510.15 138 ί A Λ3 F. xL --^N^N Ύί 8 -EM ^H h |l 1 ° 611.8,613.8 139 IZ O xC0|k. - 0*11 VS %ό IZ; About 3 φ 614.8,616.8

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140 OH 0 and 8 ⁿ n Η I ° 600.8, 602.8 142 About | ° ζΠ ^ΗΝ ___ ^OMe x%^\^s=o Ν π h H 1 ° 614.13.616.13 143 F A^N^OMe 612.11.614.11 145 '^· IZ fT - 0*11 vs 4 \ 471.07, 473.07 146 Y'o. ^4..O £ zx L 525.12, 527.12 147 o o Ύι S ^H JI JL ^s=o N H 0 1° ^^Cl 586.10.588.10 148 -f IZ I >° Ύ J ⁵ ZI th 663.14, 665.14 [M+HCO2G 149 OH Me I OH ^f y5l 1 h U 1 ° S^ci 480.10, 482.10

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151 he _{_} ___1. yun ιΓΧ'° ^''^ύϊι 516.08.518.08 152 Ά VF 0 Gauss/ HN^Ot-Bu UL-Dh JL .S=O N P F N YX O H 0 1° 704.18.706.18 154 o £ /^N^^Ph ^Ρ γ4 ⁰ \ HN^OMe JL S=ON P F'^N'YV S' O h 0 1° ^i^CI 662.13.664.13 155 it 601.14, 603.14 157 X 1 °¥ CHA iuo_ K7 O=(V ZI -ΌIL 553.15 158 xz\O— /1 O <Λ ιι<ρ _ ^A-CHO*' o X ZZ IL 553.05 159 •J ZZ x) V" NA and<° - XZZ 614.10 160 O x 7 ABOUT- /( ABOUT Member and<o _ K7 -=0-0 --0IL 558.15

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161 zz “O*·· \> $=° zz about X 565.15.567.15 162 it 1 Jk X\^OH SA about kA N h U JL ° 565.15.567.15 165 q4 >° zz \ § / ll o CL H^o _ S...U) ^X Q £ zz LL 540.30 166 .OH OH F 1 A^V' ^Me VL 1^./=74 h H 1 ° 482.20 167 /-zz I ABOUT - OL1 VS \ zz o =/ V 580.2 168 F? ^H H fA f^X £χ/'Υ^ ^ν I and ° IL xJL Ji _____ .s=o N h |PV ⁴ o 596.2 169 zz O z ^M, K - 0^11 Vs A. / I \ zz ;5 ^{l o} z 571.2 170 FJ ^H Η fJ. _o Α/^γχ ¹ ii : \l 0 ^{Me Me} ^^ ^ν, ιΓ^Γ ⁸ 'ο ^Ο Η Η 1 ⁰ 585.2 171 0-’ zz n xWM'V. - 0^11 V> A\zz A o z 583.2

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172 he he rapemicheskpn 528.10, 530.10 173 he he rapempchkpp 528.10, 530.10 176 he he TU 1 h π i ° ^i^CI 542.12, 544.12 177 V zz X ο _χ ω>··^ - Ol 1 Vs 4. / = ^X o )-o / z 542.12, 544.12 178 _F OH OH %ί^α 572.13.574.13 179 OH OH '4 Η U 1 ° 572.13.574.13 184 IZ $ O Z«|K - 0^11 VS 4, $=° IZ I I 557.15 185 he o Y1 U ^H “ ^e η Π I ° 571.25

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186 z about BH IUO _ ζχ у 571.20 187 0 ' τζ ο _ζ ί» κ ^_ Ο-'ll VS ο \ Ι Υ—ο .. X ζζ Η ζ 585.20, 587.15 188 he ο F.1 Υη i Πι ^Η J. n Π I ° ^J^ci 585.20, 587.15 189 S ο -Λ p 585.20, 587.15 190 he ο Α _ο ftMr* A ^^s=o N h Η 1 ⁰ 571.20 191 he o N s' η Π 1 ⁰ 599.30 192 z 7; zz °4 Έ· SA n<o u. 583.20 193 F __° ^H y ^F vSi ° /^fZ'^OH CchA. A^zS^fS=oN η η i ° 526.05

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194 F Η Ί R °w° 1 ϋ Λ. 11 's' XJ he ^H ILX 471.95 195 x 7. >And ϋ 571.00 196 7. >«about 571.00 197 F _f ___1 Il Ί I 0..0 Г 1\ У AI _s XJ ^H I 1 / 4A _C | ^H0 530.10 198 R 1 ^l ⁿ h ί I o O g γ-l VX I___χ XJ h (TV /“ ^A _C| HO 530.00 199 F FX OH y—OH Ύί R °h/° l^^lx 11 XX 's' XJ he 'N' ^H h J ⁴ —^Cl 502.05, 504.05 200 F R JL OH Il Ί I ° _v .° rX-^. L A and 1 / \ N Ji ^»ι Η [1 J 443.95 201 F F i <°X > ^H vS i %/Qv° ^H H ]fV'o ^H o 458.00, 460.00

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207 FX ^OH /° ^{H n} 1JL 473.95 208 P XZ O 2 /% o X 501.95 209 X o Y 501.95 211 mix 4:1 413.95 212 R XZ (G O / "'K - OCH1 V" o\u003e / o 4 X S No. 436.05 213 f 2 X ΐ ^β ><D 11-0 _ d 436.30 214 f S X-' ABOUT- /( ABOUT chd u < o _ d 434.20 215 _F OH 0 510.00

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216 P 0-- ΙΖ i οΛ 484.00 217 he r^N Ο n Η 1 ⁰ % ⁱ ^ci 436.05 218 _c OH he NY A n U 1 ° 512.05 219 F π ί 1? ° «° Г ι ^F P J P a* 1 1 ^A^/y ⁸ '^/ ⁿ 1L - XI 462.00 220 F ⁰ OO 429.95 221 He G PM η n C 1 ° 436.25 222 F Il Ί I °w° G H L A X 1 J Y- N' V'XZ'/ η Η 1 ° %i^ci 498.05 223 F ₽ 1 P ^H Al^Shu η || 1° 498.00

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224 F ^/ ^oh η Ί I 0 O 1 T'\ P P JL J )___ Η |[Ύ HO 500.05 225 0"' IZ r o ω< A 500.05, 502.05 226 0"' ΙΖ r o ω< % P 444.00 227 0 ' ΙΖ -f o w< ъ 424.00 228 0 ' IZ A A About z 514.05 229 Z O >"oh i 514.10 230 F no. 488.00 231 512.05

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232 Π ΙΖ £ Α Π 490.05 233 F f^ X. τι ο ο ο AZhd 428.00 234 F ⁰ Ο Ο __rn 488.00 235 A.-'UU' 470.00 236 π O ^- ' ζζ A οϊ ηζ, Α ъ ζ 470.00 237 Π 0ΙΖ Λ Α ⁵ X 484.05 238 F F JL ° ^Η AUD 484.05 239 F ^F yS Oo- 440.00

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240 F K Il O 0 0 YES· Oh 444.00 241 F _______L Ύχ 1 _ V and id xv° _H 7 HO 258.00 242 F OOO TDU OME 471.95 243 F SCS About OO DboA- He 458.00 244 F Ii ⁰ OO Daa O^ OMe HN—' 529.00 245 F tG η ° oo W y—OH ΗΝ—' 503.10 246 F about OO OME HN—( Me 543.00

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247 F F^XX ° O O O^ y—OH HN—ζ Me 517.15 248 F f^^L Τι X 0 0 0 \ome HN—' 'o 542.95 249 F ^F>4 T| ^Z ^I ⁰ OO y OH HN—' 517.15 250a i oL >m o % 442.05 250b Ff. jl Ti X ⁰ 0 0 W- 0 444.05 251 F r. JL Τι X ⁰ O O 0 430.05 252a F A. Il Ί I °O° U- _H X^-X Ti ^*1 'f \ ^H II ^JL x- ^OH 414.05 252b F ^F % ^X X O o o 414.05

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253a F F^ A. TZL a V hj ^H UA VA A O 499.10 253b FJL ABOUT 499.10 254 F ⁰ O O A\sXi^ 0 473.10 255 F ^F %f ^z % ° o o Shd. About Me 515.05 256a F Ύί Y o o U A AND CH' ΥΛ ^{n n} U A \Α·. - ^N ^ Г % 0 Me 485.15 256b F ^F ' ^s ii ^z ^i ° 0 0 O Me 485.15 257a F ^F %ri ^x *^^i ° 0 o ^-CI r %r o o 513.10 257b F F. .X Ti X ⁰ 0 513.15

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258 F ⁰ oo 0 487.15 260 F _______L Ti O o 0 425.95 280 & ζζ 4 “ O'll oVSl_ _D □о 7т ° ο X 292.13, 294.13 [M- Η]' 282 He ΆνΜ 564.29, 566.29 294 π Ο— ζζ if ο _χ ωι ^ “ o ^x u Vs о 1 / >=ζ ο / ζ 621.13, 623.13 [M+HCOg]· 294a ζζ if ο _χ ω> ^ θ ^χ ν> ο 1 / f~W~° / ^ζ ζ 621.13, 623.13 [M+HCO ₂ ]" 294b ζζ Ο /«· _κ θ ^χ ν> Ο 1 / / ^Ζ χ 621.13, 623.13 [M+HCO2G 295 He 621.13, 623.13 [M+HCO2G 295a ζζ ο _χ ωι >^ θ ^χ ν> ο 1 / /—ζ ^S— ο \ Vа ¹ ' ⁼ \ ζ 621.13, 623.13 [M+HCO2]

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295b Oh 621.13, 623.13 [M+NSOGG 296 Oh MJ 621.13, 623.13 [M+NSOGG 296a Oh 621.13, 623.13 [M+NSOGG 296b Oh 621.13, 623.13 [M+NSOD 297a 625.11, 627.11 [M+HCOg]' 297b & IZ A ο o ^x V> o, / / ^z -η 625.11, 627.11 [M+HCOg]' 299 He 625.11, 627.10 [M+HCO2] ^- 300 he luu cf ₃ 675.10, 677.10 [M+HCOg]' 301 θ \ Ζ X )-° o-A. 4a °/ _z o _ A ZZ IL 657.32, 659.32 [M+HCOg]'

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302 Oh A 8 HMGG0 613.24, 615.23 [M+HCO ₂ ]" 302a A IZ A O / I'K. °'ph GC z 613.24, 615.23 [M+HCO ₂ ]" 302b z o zz --0 613.24, 615.23 [M+HCO ₂ ]" 303 & ZZ A O _X W>'·^ o 1 / lip °'^b z 597.27, 599.26 [M+HCO ₂ ]" 304 0ZZ A o _χ ω< .4 "°* ¹¹ V> — 539.16, 541.16 [M- Η]' 305 he rt al ^ϊΑυ M U 0 IL JL .s=o X h |ίΎ'° 624.10, 626.10 [M+SOgG 306 F? ^H GL) _Fx A η^τ7^ ^ν Υ U ° А^з=° N h U 1 ° 553.26, 555.26 [M- Η]' 310 F ^F >iA o 0 o ? ^H 610.12, 612.12 [M+NSOGG 311 F ^F >iA o O o ° ^H 610.12, 612.12 [M+HCO ₂ ]-

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312 ^F αΑ OOO ? ⁿ O 629.11, 631.11 [M+NSOGG 313 he but 20- ^F ^lVY'i GT UAC1 659.31, 661.31 [M+NSOGG 324 OH OH FT Ϊ Ί η ^he iQp 558.14 [M+HCOgG 325 He I χ--.A___.he VS 8 Dh ΛθΑ 514.11 [M-NG 326 OH OH F Ϊ 1 f___I A/DmyA^ ⁴ - ^|| H OH DDxD A>x _z S=ON f^^nAt-4 x' H LA- 576.13 [M+HCOgG 335 _F OH OH f^JL %/Α'ΤγΧ Td iLADh ^oh h jAf ° Me^A^XI 588.30, 590.30 [M+NSOGG 338 D OH OH WF 580.15, 582.15 [M+NSOGG 342 ZZ A n _χ ω··£ 0*11 VS 567.19, 569.19 [M- Η]' 343 H o 1 A A f. JL h /ΣΎ'/ν ^nh Xaa^XV ΑμΎϊ -xwa shomer 3.5:1 548.23, 550.23 [M- Η]'

349 ZZ / O X""··*. ^_ o ^x n vA ° // o °—( ¹ Z \ I >—Z 657.11, 659.11 [M+HCO2G 350 he 1 lASD JLf ^p aD^ /ax/ α^ ^ν ' ύ | Η η T he f W-oy 657.11, 659.11 [M+HCO2G 351 he he F 11 UL ^he V n ifY'° ' S^ci 651.14, 653.14 [M+HCO2G 352 he he F ϊ Ϊ Χ^ΪΙ 8 ^V ^s <° ^{N H} ll 1 Si^ci 637.12, 639.12 [M+HCO2G

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The following examples were prepared using procedures analogous to those described above.

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biological activity.

Ways.

HepAD38 cells are stored as previously described (Ladner et al., Antimicrob. Agents Chemother. 1997, 4, 1715). Briefly, cells at confluence were transferred to DMEM/F12 medium in the presence of 10% PBS, penicillin/streptomycin, 250 g/ml G418, and 1 μg/ml tetracycline. New compounds are tested by first washing the cells three times with PBS to remove tetracycline and placing them in a 96-well plate at 35,000 cells/well. Compounds dissolved in DMSO are then diluted 1:200 into wells containing cells. Five days after adding the compound, the material is collected for analysis. For an extended 8 day assay, cells are placed and processed as described above, but medium and compound are updated at d2 and d5 after initial processing.

On the day of collection, virion DNA is obtained by lysing with stepwise lysis and stabilizing buffer and then quantified by quantitative real-time PCR. After diluting the samples according to the linear range of their respective assays, following the manufacturer's recommended protocol for the quantitation of HBsAg (Alpco) or HbeAg (US Biological) viral proteins, commercially available ELISA kits are used. Regardless of the reading, compound concentrations are given that reduce the accumulation of viral product in cell lysates or supernatants by 50% compared to no drug control (EC50); the ranges (EC ₅₀ ) are as follows: A<0.1 μM; B 0.1-1 μM; C>0.2 μM.

Compound toxicity is assessed by cell seeding at 15,000 cells/well and compound treatment as described above. Three days after compound administration, cells are treated with ATPLite reagent and compound concentrations are adjusted that reduce total ATP levels in wells by 50% compared to no drug control (CC ₅₀ ); CC ₅₀ ranges are as follows: A>25 μM; B 10-25 μM; C<10 µM.

Table 1

List of activities

Connection number HepAD38 EC ₅₀ (µM) Connection number HepAD38 EC ₅₀ (µM) 1 A 2 IN 3 A 4 A 5 A 6 A 7 A 8 WITH 9 A 10 A AND A 12 A 13 A 14 A 15 A 16 A 17 A 18 A 19 WITH 20 A 21 A 22 A 23 A 24 WITH 25 WITH 26 A 27 WITH 28 A 29 WITH thirty A 31 V 32 WITH 33 With 34 A 35 A 36 A 37 IN 38 WITH

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39 A 40 C 41 A 42 A 43 C 44 C 45 C 46 C 47 IN 48 A 49 c 50 A 51 A 52 A 53 A 54 A 55 A 56 IN 57 B 58 A 59 IN 60 IN 61 A 62 A 63 A 64 IN 65 A 66 A 67 C 68 A 69 A 70 A 71 A 72 A 73 A 74 IN 75 A 76 A 77 C 78 C 79 C 80 A 81 A 82 A 83 C 84 C 85 C 86 A 87 A 88 A 89 A 90 A 91 A 92 A 93 A 94 A 95 C 96 IN 97 IN 98 A 99 A 100 A 101 C 102 C 103 A 104 C 105 A 106 A 107 C 108 A 109 A 110 A 111 A 112 A 113 A 114 A 115 A 116 A 117 A 118 A 119 C 120 C 121 A 122 A 123 A 124 A 125 A 126 A 127 C 128 C 129 A 130 A 131 A 132 C 133 C 134 C

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135 A 136 A 137 A 138 A 139 C 140 WITH 141 IN 142 IN 143 c 144 IN 145 c 146 With 147 A 148 V 149 A 150 A 151 A 152 WITH 153 IN 154 With 155 A 156 A 157 A 158 A 159 A 160 WITH 161 A 162 A 163 A 164 A 165 A 166 A 167 A 168 A 169 IN 170 A 171 C 172 A 173 A 174 A 175 A 176 A 177 A 178 A 179 A 180 A 181 A 182 A 183 A 184 A 185 A 186 A 187 A 188 A 189 A 190 A 191 A 192 A 193 A 194 IN 195 C 196 WITH 197 C 198 IN 199 c 200 With 201 c 202 A 203 A 204 IN 205 c 206 V 207 c 208 With 209 c 210 A 211 c 212 WITH 213 c 214 With 215 A 216 V 217 c 218 A 219 c 220 WITH 221 c 222 V 223 V 224 V 225 V 226 With 227 With 228 With 229 With 230 With

- 103 041670

231 C 232 C 233 c 234 c 235 c 236 c 237 c 238 c 239 c 240 c 241 c 242 c 243 c 244 c 245 c 246 c 247 c 248 c 249 c 250a c 250b c 251 c 252a c 252b c 253a c 253b c 254 c 255 c 256a c 256b c 257a c 257b c 258 c 260 c 272 A 279 A 280 A 281 c 282 A 283 V 284 A 284a A 284b A 285 A 285a A 285b A 286 IN 287 c 288 A 289 A 290 A 291 A 291a A 291b A 292 C 293 A 294 A 294a A 294b A 295 A 296 A 296a A 296b A 297 A 298 A 299 A 300 C 301 A 302 A 303 A 304 A 305 A 306 A 307 IN 308 A 309 IN 310 C 311 WITH 312 C 313 A 314 A 315 WITH 316 C 317 A 318 C 319 A 320 A 321 A 322 A 323 IN 324 A 325 A 326 A 327 IN 328 IN 329 A 330 C 331 A 332 A 333 A 334 A 335 A 336 A 337 C 338 A 341 C 342 A 343 A 344 A 345 A 346 347 348 A 349 IN 350 A

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Claims (13)

table 2 Summary results for cytotoxicity Connection number ATPlite CC50 (µM) Connection number ATPlite CC50 (µM) 1 >3 4 >12.5 5 >3 9 >6.25 10 >12.5 eleven >6.25 12 >3 13 >6.25 36 A 39 >6.25 41 >6.25 52 A 53 >6.25 54 A 55 A 58 >12.5 61 >6.25 62 >12.5 66 IN 68 A 70 A 71 >3 72 >3 73 >6.25 88 >6.25 89 >6.25 90 >6.25 91 IN 92 >6.25 93 >6.25 111 >3 112 >6.25 113 >6.25 114 >6.25 116 >3 117 >6.25 122 A 125 >6.25 126 >6.25 129 A 130 >12.5 131 >12.5 135 WITH 136 >12.5 137 >6.25 149 A 150 A 151 A 152 A 155 A 162 >6.25 163 A 167 >12.5 168 IN 172 A 173 >6.25 176 >6.25 177 >6.25 178 A 179 A 180 A 182 A 183 A 185 >12.5 186 >12.5 187 >12.5 188 >12.5 189 >12.5 191 A 192 >12.5 193 A 215 A 218 IN 272 A 279 A 280 A 284b A 285a A 285b A 289 A 290 A 291 A 294 A 296 A 302 A 303 A 308 A 317 A 319 A 325 A 326 A 329 A 331 A 332 A 335 A 338 A 343 A 344 A 345 A While the invention is specifically illustrated and described by reference to the preferred embodiments of the invention, those skilled in the art will appreciate that numerous changes in form and detail may be made herein without departing from the scope of the invention as set forth in the appended claims. CLAIM 1. A compound of formula (XVII) or a pharmaceutically acceptable salt thereof where R ₂₁ , R ₂₁ ' and R ₂₁ are independently selected from hydrogen, fluorine, methyl, difluoromethyl and trifluoromethyl; R39 is a hydroxyl group and R40 is -[C(R ₄₁ )(R ₄₂ )]qR ₄₃ where q is 2; R ₄₁ is hydrogen and R42 is hydroxyl; or, alternatively, R _4i and R ₄₂ of the same C(R ₄ i)(R 4 ₂ ) group together form oxo; and R ₄₃ is C1- _C6 alkyl. 2. A compound according to claim 1, characterized in that R _2i ' and R ₂₁ are fluoro and R _2i is hydrogen. 3. The compound according to claim 1, characterized in that R _2i , R _2i ' and R ₂₁ are fluoro. 4. The compound according to claim 1, having the structure he he 5. The compound according to claim 1, having the structure 6. The compound according to claim 1, having the structure 7. The compound according to claim 1, having the structure he he 8. The compound according to claim 1, having the structure 9. The compound according to claim 1, having the structure he he 10. The compound according to claim 1, having the structure he he 11. Connection by π. 1 having the structure 12. A pharmaceutical composition containing a compound according to any one of claims 1-11 and a pharmaceutically acceptable carrier or excipient. 13. A method of treating or preventing hepatitis B virus infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-11. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky per., 2 - 106 -