Carboxamide derivatives and the use thereof as medicaments for the treatment of hepatitis b.

(54) Title: CARBOXAMIDE DERIVATIVES AND THEIR USE AS DRUGS FOR THE TREATMENT OF HEPATITIS B.

(54) Title: CARBOXAMIDE DERIVATIVES AND THE USE THEREOF AS MEDICAMENTS FOR THE TREATMENT OF HEPATITIS B.

(57) Summary

HBV replication inhibitors of formula (I) (see Formula) including the stereochemically isomeric forms, and salts, hydrates and solvates thereof, wherein X and from R1 to R7 have the meanings defined herein ; The present invention also relates to processes for preparing such compounds, to pharmaceutical compositions containing them and to their use, alone or in combination with other HBV inhibitors, in therapy against HBV.

(57) Abstract

Inhibitors of HBV replication of formula (I) including stereochemically isomeric forms, and salts, hydrates, solvates thereof, where X, R1 to R7 have the meaning as defined herein. The present invention also relates to processes for preparing said compounds, pharmaceutical compositions containing them and their use, alone or in combination with other HBV inhibitors, in HBV therapy.

CARBOXAMIDE DERIVATIVES AND THEIR USE AS DRUGS FOR THE TREATMENT OF HEPATITIS B

BACKGROUND OF THE INVENTION

Hepatitis B virus (HBV) is an enveloped, partially double-stranded DNA (dsDNA) virus that belongs to the Hepadnavirus family (Hepadnavirídae). Its genome contains 4 overlapping reading frames: the prenuclear / nuclear gene; the polymerase gene; the L, M and S genes, which encode the 3 envelope proteins; and gene X.

When infection occurs, the partially double-stranded DNA genome (relaxed circular DNA, cDNA) is converted to a covalently closed circular DNA (ccDNA) in the nucleus of the host cell, and the viral mRNAs are transcribed. Once encapsulated, the pregenomic RNA (pRNA), which also encodes the nuclear protein and Pol, acts as a model for reverse transcription, which regenerates the partially double-stranded DNA (cDNA) genome in the nucleocapsid.

HBV has caused epidemics in parts of Asia and Africa, and is endemic in China. HBV has infected approximately 2 billion people worldwide, of which approximately 350 million have developed chronic infections. The virus causes hepatitis B disease, and chronic infection correlates with a markedly increased risk of developing cirrhosis and hepatocellular carcinoma.

Transmission of the hepatitis B virus occurs due to exposure to infectious blood or body fluids, although viral DNA has been detected in the saliva, tears, and urine of chronic carriers with a high concentration of serum DNA.

There is an effective and well-tolerated vaccine, but direct treatment options are currently limited to interferon and the following antiviral agents: tenofovir, lamivudine, adefovir, entecavir, and telbivudine.

Likewise, heteroaryldihydropyrimidines (PAHs) were identified as a class of HBV inhibitors in animal models and tissue culture (Weber et al., Antiviral Res. 54: 6978).

Document W02013 / 006394, published on January 10, 2013, refers to sulfamoylarylamides that are active against HBV.

WO / 2013/096744, published on June 26, 2013, refers to compounds that are active against HBV.

Among the problems that direct antiviral agents against HBV can present are toxicity, mutagenicity, lack of selectivity, unsatisfactory efficacy, unsatisfactory bioavailability and difficulty in their synthesis.

There is a need for additional HBV inhibitors that can overcome at least one of these disadvantages or that have additional advantages such as higher potency or a broader safety framework.

BRIEF DESCRIPTION OF THE INVENTION

The present invention relates to a compound of formula (I)

<img file="MX2016005342A_D0001.tif" />

(I) or a stereoisomer or a tautomeric form thereof, in which:

»/ V \ [\ | I'VXZ '/

* X represents

<img file="MX2016005342A_D0002.tif" />

, Ra, Rb, Re, Rd, Re, Rf and Rg are each independently selected from the group consisting of hydrogen and methyl;

Rh is hydrogen;

Ri is hydrogen;

R¹, R² and R³are independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, -CHF₂, -CH₂F, -CF₃, -CN and methyl;

R⁶ is selected from the group consisting of alkyl CrC₆ and a 3-7 membered saturated ring optionally containing one or more heteroatoms each independently selected from the group consisting of O, S, and N, said CiQ alkyl or 3-7 membered saturated ring being optionally substituted with one or more substituents selected from the group consisting of fluoro, Ci-C alkyl₃ optionally substituted with one or more fluoro, -CN, OH;

R⁷ represents hydrogen;

or a pharmaceutically acceptable salt or a solvate thereof.

The invention further relates to a pharmaceutical composition comprising a compound of Formula (I) and a pharmaceutically acceptable carrier.

The invention also relates to compounds of formula (I) for use as a medicament, preferably for use in the prevention or treatment of an infection caused by HBV in a mammal.

In a further aspect, the invention relates to a combination of a compound of formula (I) and another HBV inhibitor.

Definitions

The expression Ci alkyl.₃ or CrC alkyl₄, as a group or part of a group, refers to a hydrocarbyl radical of Formula C_nH_{2n +}i where n is a number between 1 and 3. In the case that the alkyl Ci-₃ is coupled to another radical, it will refer to a Formula C_nH_2n. The Ci_ alkyl groups₃ they comprise 1 to 3 carbon atoms, more preferably 1 to 2 carbon atoms. Alkyl C_b3 includes all linear or branched alkyl groups with 1 to 3 carbon atoms, and therefore includes groups such as, for example, methyl, ethyl, n-propyl and ipropyl.

C ^ -alkyl, as a group or part of a group, defines straight or branched chain saturated hydrocarbon radicals containing from 1 to 4 carbon atoms such as the group defined for C-alkyl_b3 and butyl, and the like.

The alkyl Ci-₆, alkyl C₂.₆ and alkyl C₃.₆, as a group or part of a group, defines straight or branched chain saturated hydrocarbon radicals containing from 1 to 6 carbon atoms or from 2 to 6 carbon atoms or from 3 to 6 carbon atoms such as groups defined for alkyl Ci-₄ and pentyl, hexyl, 2-methylbutyl, and the like.

The term 3-7 membered saturated ring, as used herein, refers to a cyclic hydrocarbon saturated with 3, 4, 5, 6 or 7 carbon atoms and is generic for cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl or cycloalkyl C₃, CL, C₅, C₆or C7.

Such a saturated ring optionally contains one or more heteroatoms, so that at least one carbon atom is replaced by a heteroatom selected from N, O and S, in particular from N and O. Examples include oxethane, tetrahydro-2 // - pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, thiolane 1,1-dioxide and pyrrolidinyl. Cyclic hydrocarbons saturated with 3 or 4 carbon atoms and 1 oxygen atom are preferred. Examples include oxetane and tetrahydrofuranyl.

The terms halo and halogen are generic for fluoro, chloro, bromo or iodo. The preferred halogens are fluoro and chloro.

It should be noted that the positions of the radical in any molecular residue used in the definitions can be at any point of said residue, provided it is chemically stable. For example, pyridyl includes 2-pyridyl, 3-pyridyl, and 4-pyridyl; pentyl includes 1-pentyl, 2pentyl and 3-pentyl.

A link indicated with indicates the junction of the indicated fragment with respect to the main structure of the molecule.

The positions indicated on the phenyl (eg, ortho, meta, and / or para) are indicated relative to the bond that connects the phenyl to the backbone. In an example regarding the position of R¹, any location is indicated with respect to the nitrogen (*) connected to the main structure:

<img file="MX2016005342A_D0003.tif" />

(I)

When any variable (eg, halogen or Ci_alkyl₄) appears more than once in any constituent, each definition will be independent.

For therapeutic use, the salts of the compounds of Formula (I) are those in which the counterion is pharmaceutically or physiologically acceptable. However, salts having a non-pharmaceutically acceptable counterion may also be useful, for example, in the preparation or purification of a pharmaceutically acceptable compound of Formula (I). All salts, whether pharmaceutically acceptable or not, are included within the scope of the present invention.

The pharmaceutically acceptable or physiologically tolerable addition salt forms that the compounds of the present invention are capable of forming can be conveniently prepared using suitable acids such as, for example, inorganic acids such as hydrochloric acids, e.g. eg, hydrochloric or hydrobromic acid; sulfuric; hemisulfuric; nitric; phosphoric and similar acids; or organic acids such as, for example, acetic, aspartic, dodecylsulfuric, heptanoic, hexanoic, nicotinic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic, malonic, succinic, maleic, fumaric, methyl, tartaric, citric, methanesulfonic, benzenesulfonium, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and similar acids.

Conversely, such acid addition salt forms can be converted to the free base form by treating them with a suitable base.

The term salts also encompasses hydrates and solvent addition forms that the compounds of the present invention are capable of forming. Some examples of such forms are, eg. eg, hydrates, alcoholates, and the like.

The compounds herein can also exist in their tautomeric forms. For example, the tautomeric forms of amide groups (-C (= O) -NH-) are iminoalcohols (-C (OH) = N). Tautomeric forms, although not explicitly indicated in the structural formulas represented herein, are intended to be included within the scope of the present invention.

The term "stereochemically isomeric forms" of the compounds of the present invention, as used hereinabove, defines all possible compounds consisting of the same atoms linked by the same sequence of bonds, but which have different three-dimensional structures that are not they can interchange, that the compounds of the present invention can possess. Unless otherwise mentioned or indicated, the chemical designation of a compound encompasses the mixture of all possible stereochemically isomeric forms that said compound may possess. Said mixture can contain all the diastereomers and / or enantiomers of the basic molecular structure of said compound. All stereochemically isomeric forms of the compounds of the present invention, both in pure form and in admixture with one another, are intended to be encompassed within the scope of the present invention.

The pure stereoisomeric forms of the compounds and intermediates mentioned herein are defined as isomers substantially free of other enantiomeric or diastereomeric forms of the same basic molecular structure of said compounds or intermediates. In particular, the term "stereoisomerically pure" refers to compounds or intermediates that have a stereoisomeric excess of at least 80% (ie, a minimum of 90% of one isomer and a maximum of 10% of the other possible isomers. ) and up to a stereoisomeric excess of 100% (i.e. 100% of one isomer and none of the others), more specifically, compounds or intermediates having a stereoisomeric excess of from 90% to 100%, even more specifically that they have a stereoisomeric excess from 94% to 100% and, even more specifically, that they have a stereoisomeric excess from 97% to 100%. The terms enantiomerically pure and diastereomerically pure should be interpreted in a similar way, but with reference to the enantiomeric excess and the diastereomeric excess of the mixture in question, respectively.

Pure stereoisomeric forms of the compounds and intermediates of this invention can be obtained by applying procedures known in the art. For example, enantiomers can be separated from each other by selective crystallization of their diastereomeric salts with optically active acids or bases. Some examples of these are tartaric acid, dibenzoyltartaric acid, ditholuoyltartaric acid and canphosulfonic acid. Alternatively, the enantiomers can be separated by chromatographic techniques using chiral stationary phases. Such pure stereochemically isomeric forms can also be obtained from the corresponding pure stereochemically isomeric forms of suitable starting materials, provided that the reaction occurs stereospecifically. Preferably, if it is desired to obtain a specific stereoisomer, said compound will be synthesized by stereospecific preparation methods. In these methods, enantiomerically pure starting materials will be conveniently employed.

The diastereomeric forms of formula (I) can be obtained separately by conventional methods. Suitable physical separation methods that can be beneficially used are, for example, selective crystallization and chromatography, eg. eg, column chromatography.

The present invention is also intended to include all isotopes of the atoms that appear in the compounds of the present. Isotopes include those atoms that have the same atomic number but that have different mass numbers. By way of general example and without limitation, hydrogen isotopes include tritium and deuterium. Carbon isotopes include C-13 and C-14.

DETAILED DESCRIPTION OF THE INVENTION

As long as they are used hereinafter, the expressions compounds of Formula (I),

<img file="MX2016005342A_D0004.tif" />

(I) or the compounds of the present or similar expressions are intended to include the compounds of the general formula (I), (II), (III), salts, stereoisomeric forms and racemic mixtures or any subgroups thereof.

In a first aspect, the invention provides a compound of formula (I)

<img file="MX2016005342A_D0005.tif" />

or a stereoisomer or a tautomeric form thereof, in which:

/ It represents

<img file="MX2016005342A_D0006.tif" />

or, each of Ra, Rb, Re, Rd, Re, Rf and Rg is independently selected from the group consisting of hydrogen and methyl;

Rh is hydrogen;

Ri is hydrogen;

R¹, R² and R³ are independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, -CHF₂, -CH₂F, -CF_3í -CN and methyl;

R⁶ is selected from the group consisting of Ci-Q alkyl and a 3-7 membered saturated ring optionally containing one or more heteroatoms each independently selected from the group consisting of O, S, and N, said alkyl being CrC₆ or 3-7 membered saturated ring optionally substituted with one or more substituents selected from the group consisting of fluoro, CrC alkyl₃ optionally substituted with one or more fluoro, -CN, OH;

R⁷ represents hydrogen;

or a pharmaceutically acceptable salt or a solvate thereof.

In a second aspect, the invention provides a compound of formula (II)

<img file="MX2016005342A_D0007.tif" />

(II) or formula (III) (ΠΙ) or a stereoisomer or a tautomeric form thereof, in which:

n indicates an integer 1 or 2;

R¹, R² and R³are independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, -CHF₂, -CH₂F, -CF₃, -CN and methyl;

R⁴ and R⁵ they are independently selected from hydrogen or methyl;

R⁶ is selected from the group consisting of Q-Cg alkyl and a 3-7 membered saturated ring optionally containing one or more heteroatoms each independently selected from the group consisting of O, S and N, said Ci-C alkyl being₆ or 3-7 membered saturated ring optionally substituted with one or more substituents selected from the group consisting of fluoro, CrC alkyl₃ optionally substituted with one or more fluoro, -CN, OH;

R⁷ represents hydrogen;

or a pharmaceutically acceptable salt or a solvate thereof.

In a first embodiment, compounds of formula (I), (II) or (III) are provided wherein R⁶ is selected from the group consisting of alkyl CrC₆ and a 3-7 membered saturated ring optionally containing one or more heteroatoms each independently selected from the group consisting of O, S, and N, said alkyl being CrC₆ or 3-7 membered saturated ring optionally substituted with one or more substituents selected from the group consisting of fluoro, C-alkyl_t-C₃, -CN, OH.

In one embodiment, compounds of the present invention are provided wherein R¹ selected from hydrogen, fluoro, chloro, -CHF₂, -CN, -CF₃ or methyl. In a further embodiment, at least two of R¹, R² and R³ they are fluoro, chloro or bromo. In a further embodiment, R¹ it is not hydrogen.

In another embodiment, R⁴ it is methyl.

In a still further embodiment, compounds according to the invention are indicated where R⁵it contains a 3-7 membered saturated ring optionally containing an oxygen, said 3-7 membered saturated ring being optionally substituted with methyl. Preferably R⁶ it is an oxygen-containing 4- or 5-membered saturated ring, said 4- or 5-membered saturated ring being optionally substituted with methyl.

In another embodiment, R⁶ is a Ci-C alkyl₆ branched optionally substituted with one or more fluoro.

Preferred compounds according to the invention are provided in which the stereochemical configuration of an atom (*) is as follows

<img file="MX2016005342A_D0008.tif" />

Another embodiment of the present invention relates to those compounds of formula (I), (II) or (III) or any subgroup of these, as mentioned in any of the other embodiments, where one or more of the following restrictions apply :

<img file="MX2016005342A_D0009.tif" />

and R⁶ is selected from the group consisting of Ci-C alkyl₆ which is optionally substituted with one or more fluoro;

<img file="MX2016005342A_D0010.tif" />

hydrogen or fluoro.

(c) R¹ and R³ they are independently selected from the group consisting of hydrogen, fluoro, chloro, -CN and methyl.

(d) R² is hydrogen or fluoro, and R¹ and R³ they are independently selected from the group consisting of hydrogen, fluoro, chloro and -CN.

(e) R⁶ it comprises a branched C3-C6 alkyl optionally substituted with one or more fluoro, or in which R⁶ comprises a C3-C cycloalkyl₆ where said cycloalkyl C₃-C₅ is substituted with alkyl CrC₃ replaced with one or more fluoro.

Further combinations of any of the embodiments are also included within the scope of the present invention.

Preferred compounds according to the invention are compounds 1-35 or a stereoisomer or tautomeric form thereof as mentioned in Table 1.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a therapeutically or prophylactically effective amount of a compound of formula (I) as specified herein and a pharmaceutically acceptable carrier. In this context, a prophylactically effective amount is an amount sufficient to prevent HBV infection in patients who are at risk of being infected. In this context, a therapeutically effective amount is an amount sufficient to stabilize an infection caused by HBV, to reduce an infection caused by HBV, or to eradicate an infection caused by HBV in infected subjects. In a still further aspect, this invention relates to a process for preparing a pharmaceutical composition as specified herein, which comprises intimately mixing a pharmaceutically acceptable carrier with a therapeutically or prophylactically effective amount of a compound of formula (I ), as specified in this document.

Accordingly, the compounds of the present invention or any subgroup of these can be formulated into various pharmaceutical forms for administration. As suitable compositions, there may be cited all the compositions normally used to administer drugs systemically. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound is combined, optionally in the form of an addition salt, as an active ingredient, in intimate mixture with a pharmaceutically acceptable carrier, said carrier being able to take a variety of forms depending on the form of the preparation desired for administration. It is desirable that these pharmaceutical compositions adopt a unit dosage form suitable, in particular, for administration orally, rectally, percutaneously, or by parenteral injection. For example, in the preparation of compositions in an oral dosage form, any of the usual pharmaceutical media such as, for example, water, glycols, oils, alcohols and the like can be employed in the case of oral liquid preparations such as suspensions, syrups , elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents, and the like in the case of powders, lozenges, capsules, and tablets. Because they can be easily administered, tablets and capsules represent the most convenient oral unit dosage forms, in which case solid pharmaceutical carriers are employed. For parenteral compositions, the carrier will normally comprise sterile water, at least to a large extent, although it may include other ingredients, for example, to increase solubility. Injectable solutions can be prepared, for example, in which the carrier comprises saline, glucose solution or a mixture of saline and glucose. Injectable suspensions can also be prepared, in which case suitable liquid carriers, suspending agents and the like can be employed. Also included are solid form preparations that are designed to be converted, shortly before use, to liquid form preparations. In compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and / or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, where the additives do not cause any significant detrimental effect on the skin. The compounds of the present invention can also be administered by inhalation or oral insufflation, in the form of a solution, a suspension or a dry powder, using any delivery system known in the art.

It is especially convenient to formulate the aforementioned pharmaceutical compositions in unit dosage forms because of the uniformity of the dose and because they can be easily administered. The term unit dosage form, as used herein, refers to physically discrete units suitable as unit doses, where each unit contains a predetermined amount of active ingredient calculated to produce the desired therapeutic effect associated with the required pharmaceutical carrier. Some examples of such unit dosage forms are tablets (including scored or scored tablets), capsules, lozenges, suppositories, powder sachets, wafers, injectable solutions or suspensions, and the like, and segregated multiples thereof.

The compounds of formula (I) are active as inhibitors of the HBV replication cycle and can be used in the treatment and prophylaxis of an infection caused by HBV or diseases associated with HBV. The latter include progressive liver fibrosis, inflammation and necrosis causing cirrhosis, end-stage liver disease, and hepatocellular carcinoma.

Due to their antiviral properties, particularly their anti-HBV properties, the compounds of formula (I) or any subgroup of these are useful in inhibiting the HBV replication cycle, in particular in the treatment of warm-blooded animals, in particular humans, infected with HBV, and for the prophylaxis of infections caused by HBV. The present invention further relates to a method of treatment for a warm-blooded animal, in particular a human being, infected with HBV or at risk of being infected with HBV, wherein said method comprises administering a therapeutically amount effective of a compound of formula (I).

Accordingly, the compounds of formula (I), as specified herein, can be used as a medicine, in particular as a medicine to treat or prevent an infection caused by HBV. Said use as a medicine or method of treatment comprises the systemic administration, to subjects infected with HBV or to subjects susceptible to being infected with HBV, of an amount effective to combat the conditions associated with an infection caused by HBV or an effective amount for prevent an infection caused by HBV.

The present invention also relates to the use of the compounds herein in the manufacture of a medicament for the treatment or prevention of an infection caused by HBV.

In general, it is considered that an effective daily antiviral amount would be between approximately 0.01 and approximately 50 mg / kg or between approximately 0.01 and approximately 30 mg / kg of body weight. It may be appropriate to administer the required dose as two, three, four, or more subdoses at appropriate intervals throughout the day. Such subdoses may be formulated as unit dosage forms, for example, containing from about 1 to about 500 mg, or from about 1 to about 300 mg, or from about 1 to about 100 mg, or from about 2 to about 50 mg of the Active ingredient per unit dosage form.

The present invention also relates to combinations of a compound of Formula (I) or any subgroup thereof, as specified herein, with other anti-HBV agents. The term combination can refer to a product or kit containing (a) a compound of formula (I), as specified above, and (b) at least one other compound capable of treating an infection caused by HBV (which is herein referred to as anti-HBV agent), as a combined preparation for simultaneous, sequential or separate use in the treatment of HBV infections. In one embodiment, the invention relates to a combination of a compound of Formula (I) or any subgroup thereof with at least one anti-HBV agent. In a particular embodiment, the invention relates to a combination of a compound of Formula (I) or any subgroup thereof with at least two anti-HBV agents. In a particular embodiment, the invention relates to a combination of a compound of Formula (I) or any subgroup thereof with at least three anti-HBV agents. In a particular embodiment, the invention relates to a combination of a compound of Formula (I) or any subgroup thereof with at least four anti-HBV agents.

The term "anti-HBV agent" also includes compounds capable of treating an infection caused by HBV by immunomodulation. Some examples of immunomodulators are interferon-α (IFN-a), pegylated interferon-a, or innate immune system stimulants such as Toll 7 and / or 8 receptor agonists. An embodiment of the present invention relates to combinations of a compound of Formula (IA) or any subgroup thereof, as specified herein, with an immunomodulatory compound, more specifically a Toll 7 and / or 8 receptor agonist. .

The combination of previously known anti-HBV agents, such as interferon-α (IFN-a), pegylated interferon-a, 3TC, adefovir, or a combination thereof, and a compound of Formula (I) or any subgroup thereof It can be used as a medicine in a combination therapy.

Generic synthesis:

The substituents represented by R are intended to^1,2,3, R⁷ or R⁶ In this general synthesis section, include any substituent or reactive species that is suitable for transformation into any substituent R^1,2,3 or R⁶ according to the present invention without too much complication for the person skilled in the art.

A possible synthesis of compounds of general formula (I) is described in Synthesis Equation 1. An N-protected aminocarboxylic acid (where Pg is a protecting group) of general formula (IV) can be reacted selectively with an aniline of general formula (V), for example by adding aniline (V) to a mixture of compound (IV), and a coupling agent (eg HATU) in an aprotic solvent (eg dichloromethane, DMF), together with an organic base (eg triethylamine) resulting in a compound (VI). The protecting group (Pg) can subsequently be deprotected according to known methods (for example for the Boc group, deprotection involves the addition of a strong acid such as HCI. The benzyl protecting groups are removed by means of catalytic hydrogenation by means of methods known to a person skilled in the art) giving rise to the amine salt which after removal of the solvent and addition of base (for example diisopropylethylamine) can be further reacted in one step with ethyl chloroxoacetate at reduced temperature in an aprotic solvent (for example dichloromethane) to give compounds of type (VIII). The ester group of (VIII) is then hydrolyzed by known methods (eg addition of an aqueous base). In a single step, the newly formed acid is generated after lowering the pH and removing the solvent under reduced pressure. The acidic functional group is converted to an amide functional group by using a coupling agent (eg HATU) in an aprotic solvent (eg dichloromethane, DMF), together with an organic base (eg triethylamine), and amines ( IX) resulting in compounds of formula (I). Alternatively, the ester functionality in compounds (VIII) can be converted to an amide by reaction with an amine (IX) in a closed container, or optionally in the presence of lithium bis (trimethylsilyl) amide at 0 ° C in a solvent like THF.

SYNTHESIS EQUATION 1

<img file="MX2016005342A_D0011.tif" />

IV

<img file="MX2016005342A_D0012.tif" />

Pg_s

N

<img file="MX2016005342A_D0013.tif" />

SAW

<img file="MX2016005342A_D0014.tif" />

R¹

<img file="MX2016005342A_D0015.tif" />

VIII

R⁷

<img file="MX2016005342A_D0016.tif" />

Synthesis Equation 2 describes another possible synthesis of a compound of general formula I. A compound of general formula X is reacted with ethyl chloroxoacetate, resulting in a compound of general formula XI. Upon selective hydrolysis, for example in the presence of a base such as NaOH at 0 ° C in MeOH, a compound XII is formed. This compound can be coupled with an amine of general formula IX in the presence of a coupling agent (for example HATU) in an aprotic solvent (for example dichloromethane, DMF), together with an organic base (for example triethylamine). Alternatively, compound XI can be directly converted to a compound of general formula XIII by reaction with an amine IX (for example in the case that IX is equal to isopropylamine, in EtOH at 60 ° C) resulting in selective formation of a compound of formula XIII. Hydrolysis of the ester functionality of XIII results in a compound of general formula XIV, which can be coupled with an amine of general formula V, for example under the influence of a coupling agent (for example HATU) in an aprotic solvent ( for example dichloromethane, DMF), together with an organic base (for example triethylamine), resulting in the formation of a compound of general formula I.

SYNTHESIS EQUATION 2

<img file="MX2016005342A_D0017.tif" />

SYNTHESIS EQUATION 3

<img file="MX2016005342A_D0018.tif" />

A reagent of general formula XVI can be formed, starting by reacting ethyl chloroxoacetate with an amine of general formula IX, followed by hydrolysis of ester, as shown in Synthesis Equation 3. This reagent XVI can be coupled with an amine, for example obtained after deprotection of VI, in the presence of a coupling agent (for example HATU) in an aprotic solvent (for example dichloromethane, DMF), together with an organic base (for example triethylamine), resulting in a compound of general formula I.

Procedure generates! the LCMS methods

High performance liquid chromatography (HPLC) measurement was carried out using an LC pump, a diode beam (DAD) or a UV detector and a column, as specified in the respective methods. When necessary, additional detectors were included (see Table of Methods below).

The flow from the column was fed into a mass spectrometer (MS), which was configured with an ion source at atmospheric pressure. A person skilled in the art will be able to set the adjustable parameters (eg, sweep interval, residence time, etc.) to obtain ions that allow the nominal monoisotopic molecular weight of the compound (PM) to be identified. Data acquisition was carried out with the appropriate software.

The compounds are described according to their ions and experimental retention times (t_R). If not otherwise specified in the data table, the described molecular ion corresponds to [M + H]⁺ (protonated molecule) and / or [MH] '(deprotonated molecule). In the event that the compound cannot be directly ionized, the type of adduct is specified (ie, [M + NH₄]⁺, [M + HCOOy, etc.). All results were obtained with the experimental uncertainties that are usually associated with the method used.

Hereinafter, SQD stands for Single Quadrupole Detector, MSD Mass Selective Detector, RT Room Temperature, Hybrid Ethylsiloxane / Silica Bridge, DAD Diode Beam Detector, HSS High Strength Silica, Q-Tof Spectrometers time-of-flight quadrupole mass detector, CLND, chemiluminescent nitrogen detector, ELSD evaporative light scanning detector.

LCMS methods (Flow is expressed in mL / min, column temperature (T) in ° C and analysis time in minutes). The instrument used was a Waters: Acquity® UPLC® - DAD and SQD

<td>Method code</td><td>Column</td><td>Mobile phase</td><td>Gradient</td><td>Flow T col</td><td>Analysis time</td>

<td>TO</td><td>Waters: BEH C18 (1.7 pm, 2.1 x 50 mm)</td><td>A: 0.1% HCOOH + 5% CH₃OH on H₂OR B: CH₃CN</td><td>From 95% A to 0% A in 2.5 min, up to 5% A in 0.5 min</td><td> 0.8 55</td><td> 3</td>

<td>B</td><td>Waters: BEH C18 (1.7 pm, 2.1 x 50 mm)</td><td>A: CH₃COONH₄ 10 mM at 95% H₂O + 5% CH₃CN B: CH₃CN</td><td>From 95% A to 5% A in 1.3 min, it stays for 0.7 min</td><td> 0.8 55</td><td> 2</td>

<td>C</td><td>Waters: HSS T3 (1.8 pm, 2.1 x 100 mm)</td><td>A: CH3COONH4 10 mM at 95% H₂O + 5% CH₃CN B: CH₃CN</td><td>From 100% A to 5% A in 2.10 min, up to 0% A in 0.90 min, up to 5% A in 0.5 min</td><td> 0.8 55</td><td> 3.5</td>

<td>D</td><td>Waters: HSS T3 (1.8 pm, 2.1 * 100mm)</td><td>A: CH₃COONH₄ 10 mM at 95% H₂O + 5% CH₃CN B: CH3CN</td><td>From 100% A to 5% A in 2.10 min, up to 0% A in 0.90 min, up to 5% A in 0.5 min</td><td> 0.7 55</td><td> 3.5</td>

Synthesis of the compounds:

Compound _______ 1: _______ (S) -N- (3-bromo-4,5-difluorophenin-f2-oxo-2 - (((/?) - l, l, ltr¡fluoropropan-2-yl) amino^,) acetyl^,) p¡rrol¡d¡n-3-carboxam¡da

<img file="MX2016005342A_D0019.tif" />

<img file="MX2016005342A_D0020.tif" />

Step 1. Synthesis of (5) -N- (3-bromo-4.5-dfluorophenyl) pyrrolidin-3-carboxamide. N-Boc- (3S) -l-pyrrolidin-3-carboxylic acid [CAS 140148-70-5] (1 g, 4.65 mmol), 3-bromo4,5-difluoroaniline (0.96 g, 4.65 mmol) and HATU were added (2.12 g, 5.58 mmol) to CH₂CI₂ (10 mi). N, Ndiisopropylethylamine (2.4 mg, 13.9 mmol) was added and the resulting mixture was stirred at room temperature for 4 hours. The mixture was partitioned with HCI (1M, aq., 20 ml). The organic phase was separated and the solvent was removed under reduced pressure. The crude was purified by silica gel column chromatography using a gradient of heptane to ethyl acetate to give an oil. Subsequent deprotection of the Boc group in HCI (6M in isopropanol, 15 h at room temperature) gave (S) -N- (3-bromo-4,5-difluorophenyl) p! Rrolidin-3-carboxamide hydrochloride which was used as such in the next step without further purification.

Step 2. Synthesis of ethyl (S) -2- (3 - ((3-bromo-4,5-difluorophenyl) carbamoyl) pyrroyldyl-yl) -2oxoacetate. A mixture of (S) -N- (3-bromo-4,5difluorophenyl) pyrrolidin-3-carboxamide hydrochloride (1.8 g), and triethylamine (1.47 ml, 10.54 mmol) in CH was cooled₂CI₂ (20 mi) up to 0 ° C. Ethyl chloroxoxoacetate (0.65 ml, 5.8 mmol) was added dropwise to this mixture, and the reaction mixture was stirred for one hour at 0 ° C, followed by the addition of ethyl acetate (100 ml). The organic phase was washed (1M aq. HCI, NaHCO₃ ac. and brine), dried over magnesium sulfate, the solids were removed by filtration, and the solvent was removed from the filtrate under reduced pressure. The crude intermediate was used without further purification in the next step.

Step 3. (S) -2- (3 - ((3-Bromo-4,5difluorophenyl) carbamoyl) pyrrolidin-l-yl) -2-oxoacetic acid was provided after hydrolyzing the corresponding ethyl ester using sodium hydroxide in ethanol for 15 minutes at room temperature. The reaction mixture was cooled to 0 ° C. HCI (1M aq.) Was added to bring the mixture to approximately pH 2. Brine (30 ml) was added and the mixture was partitioned with ethyl acetate (3 x 50 ml). The organic phases were pooled, washed with brine (20 ml), dried with sodium sulfate, solids were removed by filtration, and the solvent was removed under reduced pressure to provide the title compound as an oil. No further purification was performed.

Step 4. Preparation of (¿) -N- (3-bromo-4,5-difluorophenyl) -l- (2-oxo-2 - (((/?) - l, l, ltr¡fluoropropan-2-¡ l) amino) acetíl) p¡rrol¡d¡n-3-carboxam¡da. A mixture of (S) -2 (3 - ((3-bromo-4,5-difluorophenyl) carbamoyl) pyrrolidin-yl) -2-oxoacetic acid (450 mg), HATU () was allowed to stir. 0.499 g, 1.31 mmol), diisopropylethylamine (463 mg, 3.58 mmol), (/?) - l, l, l-trifluoro-2-propylamine (135 mg, 1.19 mmol), and DMF (8 ml) at room temperature for 2 hours. Ethyl acetate (100 ml) was added to the reaction mixture. The organic phase was washed with 1M HCI (aq), sodium bicarbonate (sat., Aq) and brine. Solvents were removed under reduced pressure and the crude was purified by preparative reverse phase HPLC (stationary phase: RP Vydac Denali C18-10pm, 200g, 5cm, mobile phase: NH solution₄HCO₃ 0.25% in water, CH₃CN). The desired fractions were pooled and the solvent was removed under reduced pressure to obtain compound 1 as a white solid. Method A, Rt = 1.63 min, m / z = 470.0 (MH) ', exact mass: 471.0,^XH NMR (400 MHz, DMSO-o ^) δ ppm 1.30 (d,

J = 7.0 Hz, 3 H), 1.97 - 2.31 (m, 2 H), 3.10 - 3.27 (m, 1 H), 3.39 - 3.96 (m, 4 H), 4.51 - 4.75 (m, 1 H), 7.57 - 7.80 (m, 2H), 9.26 (bs, 1H), 10.41 (bs, 1H)

Compound 2: (jN- (3-bromo-4,5-dfluorophenyl) -l- (2 - ((3-methyloxetan-3-yl) amino) -2oxoacetillpyrrolidin-3-carboxamide.

<img file="MX2016005342A_D0021.tif" />

<img file="MX2016005342A_D0022.tif" />

Compound 2 was prepared according to the method described for compound 1 with the exception that, in step 4, 3-methyloxetan-3-amine was used instead of (/?) - l, l, l-trifluoro-2-propylamine . Method A, Rt = 1.44 min, m / z = 444.0 (MH), exact mass: 445.0. 'H NMR (400 MHz, DMSO-í /₆) δ ppm 1.46 - 1.57 (m, 6H), 1.92 - 2.32 (m, 4H), 3.08 - 3.24 (m, 2H), 3.43 (dt, 7 = 12.3, 7.5 Hz, 1H), 3.49 - 3.61 (m, 2H), 3.62 - 3.77 (m, 2H), 3.78 - 3.90 (m, 2H), 3.99 (dd, 7 = 11.8, 7.6 Hz, 1H), 4.25 - 4.37 (m, 4 H), 4.58 - 4.70 (m, 4H), 7.55 - 7.86 (m, 4H), 9.18 (bs, 2H), 10.40 (bs, 2H), as a mixture of rotamers.

Compound ______ 3; ______ (5) -N- (3-bromo-4.5-d¡fluorophen¡nl- (2- (tert-butylamino) -2oxoacetyl) pyrroyldin-3-carboxamide

<img file="MX2016005342A_D0023.tif" />

<img file="MX2016005342A_D0024.tif" />

Compound 3 was prepared according to the method described for compound 1 with the exception that, in step four, 2-methylpropan-2-amine was used instead of (/?) -L, l, ltrifluoro-2-propylamine . Method A, Rt = 1.63 min, m / z = 430.0 (MH)^-, Exact mass: 431.1. NMR (400 MHz, DMSO-ok) δ ppm 1.24 - 1.36 (m, 9H), 1.91 - 2.29 (m, 2H), 3.06 - 3.25 (m, 1H), 3.37 4.01 (m, 4H), 7.60 - 7.80 (m, 2H), 7.96 - 8.03 (m, 1H), 10.39 (bs, 1H).

Compound _________ 4: __________ (35) -N- (4-fluoro-3-methylphenyl) -l- {r (l-met¡let¡l) amino1 ίθΧθ) 306ϋΙ&gt; ρίΓΓθΙίΰίη-3- € 8Γί) ΟΧ8ΓΤ1ί ( ΐ3

<img file="MX2016005342A_D0025.tif" />

<img file="MX2016005342A_D0026.tif" />

Step 1. Preparation of (S) -3 - ((4-fluoro-3-methylphenyl) carbamo, DPrrol, din-l-carboxylate tere-butyl. N-Boc (3S acid) was added ) -l-pyrrolidin-3-carboxylic CAS [140148-70-5] (20 g, 92.9 mmol), 4-fluoro-3-methylaniline (11.63 g, 92.9 mmol), and N, / V-diisopropylethylamine ( 48 mi, 279 mmol) to CH₂CI₂ (300 ml) at room temperature. HATU (42.4 g, 111.5 mmol) was added in small portions and the resulting mixture was stirred at room temperature for 15 hours. The mixture was partitioned with HCI (1M, aq, 20 ml). The organic phase was separated and the solvent was removed under reduced pressure. The crude was purified by silica gel column chromatography using a gradient of heptane to ethyl acetate to give an oil. Subsequent deprotection of the Boc group in HCI (6M in isopropanol, 15 hours at room temperature) gave (S) -N- (4fluoro-3-methylphenyl) prrolidin-3-carboxamide hydrochloride that was used as such in the next step without further purification.

Step 2. Ethyl (S) -2- (3 - ((4-fluoro-3-methlfen¡DcarbamoiDpirrol¡dl-yl) -2oxoacetate. A mixture of (S) - hydrochloride was cooled. N- (4-fluoro-3-methylphenyl) pyrrolidin-3carboxamide (0.5 g), and triethylamine (587 ml, 5.80 mmol) in CH₂CI₂ (10 mi) up to 0 ° C. Ethyl chloroxoxoacetate (290 ml, 2.13 mmol) was added dropwise to this mixture, and the reaction mixture was stirred for one hour and 20 minutes at 0 ° C, followed by the addition of ethyl acetate. The organic phase was washed (1M aq. HCI, NaHCO₃ ac. and brine), dried over magnesium sulfate, the solids were removed by filtration, and the solvent was removed from the filtrate under reduced pressure. The crude intermediate was used without further purification in the next step.

Step 3. Preparation of (35) -N- (4-fluoro-3-methylphenyl ') -l- {r (l-metylletyl) aminol (oxo) acetyl&gt; pyrroxydin-3-carboxamide . Ethyl (S) -2- (3 - ((4-fluoro-3-methylphenyl) carbamoyl) pyrrolidinl-yl) -2-oxoacetate (300 mg) was dissolved in ethanol (8 ml) and to this was added isopropylamine ( 211 mg, 3.58 mmol) as a solution in ethanol (2 ml). After 3 hours, isopropylamine (1 ml, 11.64 mmol) was added. The reaction mixture was stirred at room temperature in a closed container for 3 days. Solvents were removed under reduced pressure and the crude was purified by preparative HPLC (stationary phase: RP Vydac Denali C18, 10 pm, 200 g, 5 cm), mobile phase: 0.25% NH4HCO3 solution in water, CH₃CN). Fractions were pooled and solvents were removed under reduced pressure to obtain compound 4 as a white solid. Method A, Rt = 1.35 min, m / z = 336.4 (M + H)⁺, exact mass: 335.2. ^XH NMR (400 MHz, DMSO-¿/₆) δ ppm 1.02 - 1.16 (m, 12H), 1.93 - 2.20 (m, 4H), 2.18 - 2.22 (m, 6H), 3.04 - 3.24 (m, 2H), 3.40 (dt, X = 12.1 , 7.7 Hz, 1 H), 3.48 - 3.60 (m, 2 H), 3.60 - 3.72 (m, 2 H), 3.73 - 3.85 (m, 2 H), 3.85 - 4.01 (m, 3 H), 6.97 - 7.14 (m, 2H), 7.33 - 7.43 (m, 2H), 7.46 - 7.61 (m, 2H), 8.44 (s, 1H), 8.46 (s, 1H), 10.02 (s, 1H ), 10.05 (s, 1H), as a mixture of rotamers. Differential scanning calorimetry (from 30 to 300 ° C at 10 ° C / min), peak: 137.99 ° C.

Compound ________ 5; ________ (S) -l- (2- (c¡cloDent¡lam¡no) -2-oxoacet¡0-N- (4-fluoro-3met¡lfen¡l) p¡rrol¡d¡n-3 -carboxamida.

<img file="MX2016005342A_D0027.tif" />

Compound 5 was prepared according to the method described for compound 4 with the exception that in step 3, cyclopentylamine (10 eq.) Was used instead of isopropylamine and the duration of the reaction at room temperature was two days instead of three. Method A, Rt = 1.49 min, m / z = 362.1 (M + H)⁺, exact mass: 361.2. 'H NMR (400 MHz, DMSO-c /₆) δ ppm 1.37 1.56 (m, 7H), 1.57 - 1.72 (m, 4H), 1.75 - 1.89 (m, 4H), 1.96 - 2.20 (m, 5H), 2.18 - 2.23 (m, 6H ), 3.03 - 3.25 (m, 2H), 3.34 - 3.45 (m, 1H), 3.48 - 3.59 (m, 2H), 3.60 - 3.70 (m, 2H), 3.71 - 3.83 (m, 2H ), 3.87 - 3.97 (m, 1H), 3.97 - 4.11 (m, 2H), 6.99 - 7.13 (m, 2H), 7.38 (dd, .7 = 8.1, 3.7 Hz, 2H), 7.47 - 7.59 (m, 2H), 8.52 (s, 1H), 8.54 (s, 1H), 10.03 (s, 1H), 10.05 (s, 1H), as a mixture of rotamers. Differential scanning calorimetry (from 30 to 300 ° C at 10 ° C / min), peak: 163.50 ° C.

Compound 6: (S) -N- (4-fluoro-3-methylphenyl) -l- (2 - (((/?) - lh¡drox¡propan-2-yl) amino) -2oxoacetyl) p¡rrol¡d¡n-3-carboxamide.

<img file="MX2016005342A_D0028.tif" />

Compound 6 was prepared according to the method described for compound 4 with the exception that in step 3, (R) -2-aminopropanol (10 eq.) Was used instead of isopropylamine and the duration of the reaction at room temperature it was two days instead of three. Method A, Rt 1.14 min, m / z = 352.0 (M + H)⁺, exact mass: 351.2. ^XH NMR (400 MHz, DMSO-ofe) δ ppm 1.06 (d, 7-6.6 Hz, 6 H), 1.93 - 2.15 (m, 3 H), 2.18 - 2.22 (m, 6 H), 3.07 - 3.18 (m , 3 H), 3.26 - 3.30 (m, 1 H), 3.32 - 3.46 (m, 4 H), 3.49 - 3.61 (m, 2 H), 3.61 - 3.75 (m, 2 H), 3.76 - 3.90 (m , 4 H), 3.99 (dd, 7 = 11.7, 7.7 Hz, 1 H), 4.67 - 4.80 (m, 2 H), 7.00 - 7.11 (m, 2 H), 7.31 - 7.45 (m, 2 H), 7.46 - 7.58 (m, 2H), 8.29 (s, 1H), 8.31 (s, 1H), 10.03 (s, 1H), 10.05 (s, 1H), as a mixture of rotamers.

Compound 7: ('3S) -N-f4-fluoro-3-methylfeniD-l- {í (3-methyloxetan-3-inamino1ioxo) acetyl} pyrrolidin-3-carboxamide

<img file="MX2016005342A_D0029.tif" />

<img file="MX2016005342A_D0030.tif" />

Compound 7 was prepared according to the method described for compound 4 with the exception that in step 3, 3-methyloxetan-3-amine (2 eq.) Was used instead of isopropylamine. The reaction took place at 50 ° C for 1 week instead of at room temperature for three days as described for compound 4. Method B, Rt = 0.73 min, m / z = 364.4 (M + H)⁺, exact mass: 363.2. 'H NMR (400 MHz, DMSO- &lt;7₆) δ ppm 1.49 - 1.56 (m, 6H), 1.93 - 2.22 (m, 5H), 2.19 -

2.21 (m, 6H), 3.07 - 3.25 (m, 2H), 3.37 - 3.47 (m, 2H), 3.50 - 3.60 (m, 2H), 3.62 - 3.75 (m, 2H), 3.76 - 3.89 (m, 2H), 3.98 (dd, 7 = 11.6, 7.6 Hz, 1H), 4.27 - 4.35 (m, 4H), 4.60 - 4.70 (m, 4H), 7.01 - 7.11 (m, 1 H), 7.35 - 7.45 (m, 1H), 7.49 - 7.57 (m, 2H), 9.20 (bs, 1H), 9.25 (s, 1H), 10.10 (bs, 1H), 10.12 (s , 1 H), as a mixture of rotamers.

Compound _______________ 8; _______________ (3S) -N- (4-fluoro-3-methylphenyl) -lr {laugh! /?) -! - met¡lprop¡llam¡no} xoxo) acet¡llp¡rrolid¡n-3-carboxam gives

HN

OR

Compound 8 was prepared according to the method described for compound 4, with the exception that in step 3, (/?) -Butane-2-amine (2 eq.) Was used instead of isopropylamine. The reaction duration at room temperature was 18 hours instead of three days as described for compound 4. Method B, Rt = 0.87 min, m / z = 348.2 (MH), exact mass: 349.2.^XH NMR (400 MHz, DMSO-J₆) δ ppm 0.77 - 0.87 (m, 6H), 1.05 - 1.10 (m, 6H), 1.37 - 1.55 (m, 4H), 1.93 - 2.27 (m, 4H), 2.19 - 2.22 (m, 6 H), 3.07 - 3.26 (m, 2H), 3.37 - 3.46 (m, 1H), 3.49 - 3.60 (m, 2H), 3.62 - 3.86 (m, 6H), 3.96 (dd, J = 11.7 , 7.7 Hz, 1 H), 7.02 - 7.11 (m, 2 H), 7.35 - 7.44 (m, 2 H), 7.49 - 7.56 (m, 2 H), 8.38 (s, 1 H), 8.40 (s, 1H), 10.03 (s, 1H), 10.06 (s, 1H), as a mixture of rotamers.

Compound 9: (3S) -N- (4-fluoro-3-methylphenyl) -l- &lt;oxorf35) -tetrahydrofuran-3ylaminolacetylpyrrolidin-3-carboxamide

Compound 9 was prepared according to the method described for compound 4, except that in step 3, (5) -tetrahydrofuran-3-amine (2 eq.) Was used instead of isopropylamine. The reaction took place at 50 ° C for 2.5 days instead of at room temperature for three days as described for compound 4. Method B, Rt = 0.72 min, m / z = 364.1 (M + H)⁺, exact mass: 363.2. ^JH NMR (400 MHz, DMSO- &lt;7₆) δ ppm 1.80 - 1.91 (m, 2H), 1.96 - 2.26 (m, 6H), 2.19 - 2.21 (m, 6H), 3.07 - 3.23 (m, 2H), 3.36 - 3.45 (m, 1 H), 3.47 - 3.59 (m, 4H), 3.61 - 3.73 (m, 4H), 3.74 - 3.85 (m, 6H), 3.93 (dd, 7 = 11.4, 7.7 Hz, 1H), 4.20 - 4.35 (m, 2H), 7.01 7.12 (m, 2H), 7.33 - 7.45 (m, 2H), 7.47 - 7.57 (m, 2H), 8.80 (s, 1H), 8.82 (s, 1 H), 10.03 (s, 1H),

10.05 (s, 1H), as a mixture of rotamers.

Compound 10: (25, 35) -N- (4-fluoro-3-methylphenyl) -2-methyl-l- {T (3-methylloxetan-3Íl) amínoKOXO) acetíl'Í- P¡rrolÍd¡n-3-CarbOXamÍda

<img file="MX2016005342A_D0031.tif" />

Step 1. Preparation of methyl (S) -2-methyl- (l-phenylet, D-4,5-dihydro-IH-prolol-3-carboxylate) The title compound was prepared according to methods provided in Tetrahedron Letters, vol. 33, No. 30, pp. 4311-4312, 1992 and the references cited therein.

Step 2. Preparation of methyl ί25,35) -2-ΠΊ6ϋΙ-1 - ((5) -1-ί6ηίΐ6ϋΙ) ρίπΌΐκΙίη-3-θ3Γ0οχίΐ3ΐο. Acetic acid (2.07 ml, 36.2 mmol) was added to a solution of methyl (S) -2-methyl-l- (l-phenylethyl) 4,5-dihydro-lH-pyrrole-3-carboxylate (5.92 g, 24.1 mmol) in acetonitrile (190 ml). The reaction mixture was cooled to 0 ° C, then sodium triacetoxyborohydride (7.67g, 36.17mmol) was added and stirring was continued at 0 ° C for 3 hours. The solvent was removed under reduced pressure, the crude was reconstituted in CH₂CI₂ and Na was added₂COj (sat., Ac.). The mixture was vigorously stirred. The organic phase was removed, washed with water, and then dried over magnesium sulfate. Solids were removed by filtration and the solvent was removed from the filtrate under reduced pressure. The obtained crude oil was purified by silica gel column chromatography using a heptane / ethyl acetate gradient (from 100/0 to 70/30). The best fractions were pooled and solvents were removed under reduced pressure. The oil was triturated in heptane to give a white solid, (2S, 3S) -2-methyl - ((S) -l-phenyletyl) p-roll-din-3-carboxylate. Method C, Rt = 1.75 min, m / z = 248.4 (M + H)⁺, exact mass: 247.2. ^XH NMR (chloroform - &lt;/) conforms to the data described in Tetrahedron Letters, vol. 33, No. 30, p. 4311-4312, 1992.

Step 3. Preparation of (2S, 3S^,) -2-met¡ll- (^,('S) -l-phenyletyl') lithium pinOl¡din-3-carboxylate (2S, 3S) -2-methyl-l - (((S) -l-phenylethyl) pyrrolidin-3- methyl carboxylate (100mg, 0.40mmol) in THF (1.2ml). To this, lithium hydroxide (14 mg, 0.61 mmol) in distilled water (200 µl) and methanol (50 µΙ) was added and the mixture became clear. The resulting mixture was stirred for 18 hours. Solvent was removed under reduced pressure and the residue was used without further purification in the next step.

Step 4. Preparation of (2S, 3S) -N- (4-fluoro-3-methylfenl) -2-methyl-l- (YS) -lfenylletl) p¡rrolidin-3-carboxamide . 4-Fluoro-3-methylaniline (253 mg, 2.02 mmol) was added to a mixture of (2S, 3S) -2-methyl-l - ((S) -l-phenylethyl) pyrroyl-3-carbox Lithium tin (472 mg), HATU (1.15 g, 3.03 mmol), and Ν, Ν-diisopropylethylamine (0.7 ml, 4.04 mmol) in CH₂CÍ2. The mixture was stirred at room temperature for 1 hour. The solution was diluted in CH₂CI₂ and water, the organic phase was removed, dried over MgSO₄ and the solids were removed by filtration. The solvent was removed under reduced pressure and the crude was purified by chromatography on silica gel using a heptane / ethyl acetate gradient (from 100/0 to 70/30). The best fractions were pooled and the solvent was removed under reduced pressure to give a white solid, (2S, 3S) -N- (4-fluoro-3methylpheniO ^ -methyl-l- ^ Sj-l-phenylethiopinOlidin-S-carboxamide. Method C, Rt = 1.87 min, m / z = 341.2 (M + H)⁺, exact mass: 340.2. ^JH NMR (360 MHz, CHLOROFORM- ^) δ ppm 1.26 (d, J = 6.6 Hz, 3 H), 1.36 (d, -7 = 7.0 Hz, 3 H), 1.82 - 1.97 (m, 1 H), 2.02 - 2.18 (m, 1H), 2.26 (d, J = 1.8 Hz, 3H), 2.56 2.73 (m, 2H), 2.76 - 2.88 (m, 1H), 2.88 - 2.99 (m, 1H) , 4.08 - 4.25 (m, 1H), 6.85 - 6.98 (m, 1H),

7.22 - 7.45 (m, 7H), 9.52 (bs, 1H)

Step 5. Preparation of (2S, 3S) -N- (4-fluoro-3-methylphenyl) -2-methylpyrrolidin-3carboxamide. 10% Pd / C (123 mg) was added to a solution containing (2S, 3S) -N- (4-fluoro-3methylphenyl) -2-methyl - ((S) -l-phenylethyl) prolrol-3-carboxamide (395 mg, 1.16 mmol) in methanol (20 ml) under a nitrogen atmosphere. The reaction mixture was placed under a nitrogen atmosphere and stirred for 24 hours. Hydrogen was removed, the reaction mixture was filtered through decalite, and the residue was concentrated under reduced pressure to give a clear oil which was used without further purification in the next step.

Step 6. Preparation of ethyl 2- (í2S, 3S) -3 - (((4-fluoro-3-methylphenylDcarbamoyl) -2methylpyrrolidyl-yl) -2-oxoacetate. Ethyloxalyl chloride (0.23 ml, 2.06 mmol) is added dropwise to a solution of (2S, 3S) -N- (4-fluoro-3-methylphenyl) -2-methylpyrrolidin-3-carboxamide (244 mg, 1.03 mmol) and diisopropylethylamine (0.71 ml, 4.12 mmol) in CH₂CI₂ anhydrous (10 ml) under nitrogen atmosphere at room temperature. The reaction mixture was stirred at room temperature overnight. HCI (0.5M, aq.) Was added to the reaction mixture. The organic phase was removed, washed with NaHCO₃ (ac., sat.) and brine, dried over Na₂SW₄Solids were removed by filtration and solvents were removed from the filtrate under reduced pressure. The residue was purified by silica gel column chromatography using a heptane / ethyl acetate gradient (100/0 to 30/70) to give the title compound as an oil which was dried in vacuo at 50 ° C for 2 hours and used without further purification.

Step 7. Preparation of 2 - ((2S, 3S) -3 - ((4-fluoro-3-methylphenyl) carbamo¡0-2met¡lp¡rrol¡din-l-yl) -2-oxoacetic acid. dropwise NaOH (1M aq., 1.82 ml) to a solution of

2- ((2S, 3S) -3 - ((4-fluoro-3-methylphenyl) carbamoyl) -2-metylprolidel-l) -2-oxoacetate (204 mg, 0.61 mmol) in ethanol (5 ml). The reaction was stirred at room temperature for 2 hours, then diluted in CH₂CI₂ and water. The phases were separated and the aqueous phase was acidified with HCI (1 M aq.), The acid precipitated and was reconstituted in CH₂CI₂. The aqueous phase was extracted with CH₂CI₂. The combined organic phases were dried over Na₂SW₄Solids were removed by filtration and the solvent was removed from the filtrate under reduced pressure to provide the title compound. Method C, Rt = 1.02 min, m / z = 307.0 (MH) ', exact mass: 308.1.

Step 8. Preparation of (2S, 3S) -N- (4-fluoro-3-methylphenyl) -2-methyl-l-ir (3-methylhexetan-

3- l) Aminol (oxo) acetyl&gt; prrolidin-3-carboxamide. 3-Methyloxetan-3-amine (36 mg, 0.42 mmol) was added to a solution of 2 - ((2S, 3S) -3 - ((4-fluoro-3-methylphenyl) carbamoyl) -2-methylpyrrole ¡Din-l-il) 2-oxoacetic (128 mg, 0.42 mmol), HATU (236.79 mg, 1.5 eq) and DIPEA (145 μΙ, 2 eq) in CH₂CI₂ (5 ml) and the reaction mixture was stirred overnight at room temperature. CH added₂CI₂ and HCI (1M, aq.) to the reaction mixture. The phases were separated and the organic phase was washed with NaHCO₃(sat., ac.) and brine. The combined organic phases were dried over Na₂SW₄Solids were removed by filtration and the filtrate was concentrated under reduced pressure. The crude was purified by preparative HPLC (stationary phase: RP X-Bridge Prep C18 OBD-10 pm, 30 x 150 mm); mobile phase: NH solution₄HCO₃ 0.25% in water, CH₃CN). The best fractions were pooled and the solvent was removed under reduced pressure to obtain compound 10.

Method C, Rt = 1.46 min, m / z = 376.0 (MH) ', exact mass: 377.2. ^XH NMR (400 MHz, DMSO-cfe) δ ppm 0.99 - 1.05 (m, 6 H), 1.53 (m, J = 4.2 Hz, 6 H), 1.86 - 2.05 (m, 2 H), 2.18 -

2.23 (m, 6H), 2.25 - 2.36 (m, 2H), 3.02 - 3.23 (m, 2H), 3.38 - 3.70 (m, 3H), 3.83 - 3.95 (m, 1H), 4.27 - 4.35 (m, 4H), 4.46 - 4.57 (m, 1H), 4.60 - 4.66 (m, 4H), 4.81 - 4.94 (m, 1H), 6.99 - 7.12 (m, 2H), 7.33 - 7.42 (m, 2H), 7.45 - 7.55 (m, 2H), 9.17 (s, 1H), 9.26 (s, 1H), 9.94 (s, 1H), 10.00 (s, 1H), as a 1/1 mixture of rotamers.

Compound ________ 11: ________ (S) -N- (3-chloro-4,5-d¡fluorophenin-l- (2-oxo-2-ff (R) -l, l, ltr¡fluoropropan-2-¡l) am No) acet¡npirrol¡d¡n-3-carboxam¡da

<img file="MX2016005342A_D0032.tif" />

Compound 11 was prepared according to the method described for compound 1, step one, with the exception that 3-chloro-4,5-difluoroaniline was used instead of 3-bromo-4,5difluoroaniline. The coupling reaction to give the title compound was performed according to the procedure described for compound 13, step two, with the exception that (/?) L, l, l-trifluoro-2-propylamine was used instead of l - (trifluoromethyl) -cyclopropanamine. Method B, Rt = 1.02 min, m / z = 426.1 (MH), exact mass: 427.1. NMR (400 MHz, DMSO-cfc) δ ppm 1.30 (d, J = 7.0 Hz, 3H) 1.98 - 2.28 (m, 2H) 3.07 - 3.27 (m, 1H) 3.41 - 4.04 (m, 4H) 4.54 - 4.75 (m, 1H) 7.46 7.72 (m, 2H) 9.17 - 9.33 (m, 1H) 10.43 (m, 1H), as a mixture of rotamers.

Compound _________ 12: _________ (3S) -N- (4-fluoro-3-met¡lfenil) -l- (rfl-met¡let¡0am¡no1 (oxo) acet¡l&gt; p¡perid¡n-3-carboxam¡ gives

<img file="MX2016005342A_D0033.tif" />

Step 1. Preparation of tere-butyl (5) -3 - ((4-fluoro-3-methylphenylD carbamoyl) piperidin-l-carboxylate. A mixture of (S) -l-boc-piperidin- acid was cooled. 3-carboxylic CAS [88495-54-9] (9 g, 39.3 mmol), 4-fluoro-3-methylaniline (4.91 g, 39.3 mmol) and CH2CI2 (90 ml) up to 0 ° C followed by the addition of diisopropylethylamine (20.5 ml, 117.8 mmol) and HATU (17.9 g, 47.1 mmol). The reaction mixture was stirred at 0 ° C for 2 hours followed by the addition of citric acid (sat., Aq., 100 ml), NaHCÜ3 (sat., Aq., 100 ml), and brine. The organic phase was dried over Na₂SO4, solids were removed by filtration, and solvents were removed under reduced pressure. The crude was purified using a gradient of petroleum ether / ethyl acetate (from 100/1 to 3/1). The best fractions were pooled and the solvent was removed under reduced pressure.^JH NMR (400 MHz, CHLOROFORMS) δ ppm 1.26 - 1.37 (m, 1 H), 1.39 (s, 9 H), 1.59 (qd, 7 = 12.1, 3.4 Hz, 1 H), 1.69 (d, 7 = 13.2 Hz, 1 H), 1.91 (d, 7 = 12.6 Hz, 1 H), 2.19 (d, 7 = 1.8 Hz, 3 H), 2.40 (tt, 7 = 11.0, 3.7 Hz, 1 H),

2.75 (t, 7 = 11.7 Hz, 1H), 2.97 (bs, 1H), 3.86 (d,&gt; = 13.1Hz, 1H), 4.03 (bs, 1H), 7.05 (t, 7 = 9.3Hz , 1 H), 7.31 - 7.42 (m, 1 H), 7.51 (dd, 7 = 7.0, 2.3 Hz, 1 H), 9.97 (s, 1 H)

Subsequent deprotection of the Boc group was possible by adding CH₂CI₂(100 ml) and HCI (100 ml, in dioxane) at room temperature for 24 hours to give the intermediate (5) -N- (4-fluoro-3-methylphenyl) piperidin-3-carboxamide.

^JH NMR (400 MHz, CHLOROFORM- ^ δ ppm 1.49 - 1.87 (m, 3H), 1.95 - 2.08 (m, 1H), 2.19 (d,&gt; = 2.0 Hz, 3H), 2.80 - 2.93 (m, 2 H), 3.00 (q, 7 = 10.4 Hz, 1 H), 3.17 (d, 7 = 12.0 Hz, 1 H), 3.29 (d, 7 = 11.0 Hz, 1 H), 7.07 (t, 7 = 9.2 Hz, 1 H), 7.35 - 7.45 (m, 1 H), 7.52 (dd, 7 = 7.0, 2.3 Hz, 1 H), 8.90 (d, 7 = 11.2 Hz, 1 H), 9.12 (m, 7 = 9.5 Hz, 1H), 10.31 (s, 1H)

Step 2. The preparation of compound 12 followed procedures analogous to step 2 of synthesis of compound 4 with the exception that (S) -N- (4-fluoro-3methylphenyl) p¡per¡d¡n hydrochloride was used. -3-carboxyamide in reaction with ethyl chloroxoacetate instead of (S) -N- (4-fluoro-3-methylphenyl) pyrrolidin-3-carboxamide hydrochloride. Then, as in step three below in the method described for compound 4, isopropylamine was used in a closed container to give compound 12. Method C, Rt = 1.47 min, m / z = 350.2 (M + H)⁺, exact mass: 349.2. * H NMR (400 MHz, DMSO-ok) δ ppm 1.03 - 1.12 (m, 12 H) 1.30 - 1.52 (m, 2 H) 1.60 - 1.71 (m, 2 H) 1.71 - 1.81 (m, 2 H) 1.92 - 2.09 (m, 2H) 2.17 - 2.21 (m, 6H) 2.38 - 2.46 (m, 1H) 2.53 - 2.58 (m, 1H) 2.69 2.81 (m, 2H) 3.03 (t, 7 = 11.5 Hz, 1 H) 3.26 (dd, 7 = 13.3, 10.5 Hz, 1 H) 3.68 (d, 7 = 13.3 Hz, 1 H) 3.77 (d, 7 = 13.3 Hz, 1 H) 3.83 - 3.96 (m, 2 H) 4.18 (d, 7 = 12.9 Hz, 1 H) 4.36 (d, 7 = 12.9 Hz, 1 H) 7.02 7.09 (m, 2 H) 7.33 - 7.44 (m, 2 H) 7.50 (d, 7 = 6.9 Hz, 2H) 8.47 - 8.58 (m, 2H) 9.96 (s, 2H), a mixture of rotamers.

Compound ___________ 13: ___________ (S) -N- (3-chloro-4,5-difluorophenin-l- (2-oxo-2 - ((ltrifluoromethylkylpropropinoaminoacetylpyrrolidin-S-carboxamide

<img file="MX2016005342A_D0034.tif" />

Step 1. Preparation of (S) -3 - ((3-chloro-4,5-difluorophenyl) carbamo, d-pyrrolidin-l-carboxylate t-Butyl compound was prepared according to the procedure in step 1 of compound 1 with the exception that 3-chloro-4,5-difluoroaniline was used instead of 3-bromo-4,529 difluoroaniline, then deprotection of the Boc group and reaction with ethyl chloroxoacetate were carried out according to the described methods .

Step 2. Preparation of (S) -N- (3-chloro-4,5-d¡fluorofen¡ll- (2-oxo-2 - ((l-ftr¡fluoromet¡nc¡clopropiOamino)) acetiDpirrolid¡n -3-carboxamida · A solution of (S) -2- (3 - ((3-chloro-4,5-difluorophenyl) carbamoyl) pyrrolidin-l-yl) -2-oxoacetic acid (0.33 g, 0.99 mmol) was cooled ) in DMF (10 ml) up to 5 ° C. Then diisopropylethylamine (0.513 ml, 2.98 mmol) and l- (trifluoromethyl) cyclopropanamine (0.092 ml, 0.992 mmol) were added and stirred at 5 ° C. A solution of HATU (0.414 g, 1.091 mmol) in DMF (2 ml) was added dropwise at 5 ° C. The solution was stirred at 5 ° C for 1 hr. The reaction was quenched with water and neutralized with HCI (1M, aq.), Brine (15 ml) was added, and the compound was extracted with ethyl acetate. The organic phase was removed, dried with MgSO₄Solids were removed by filtration and solvents were removed under reduced pressure to give a solid. Solid dissolved in CH₃CN with heat and cooled to room temperature. The precipitate was removed by filtration and the filtrate was concentrated under reduced pressure. The crude was purified by flash column chromatography on silica using a heptane / ethyl acetate gradient (30/70 to 0/100). The desired fractions were collected and evaporated to dryness to give compound 13 as a white solid. Method B, Rt = 1.02 min, m / z = 438.1 (MH) ', exact mass: 439.1.^XH NMR (400 MHz, DMSO-tfc) δ ppm 1.04 - 1.13 (m, 2H) 1.22 - 1.31 (m, 2H) 1.97 - 2.27 (m, 2H) 3.09 - 3.24 (m, 1H) 3.36 - 4.00 (m, 4H) 7.49 - 7.72 (m, 2H) 9.44 (s, 1H) 10.43 (bs, 1H), as a mixture of rotamers.

Compound 14: (S) -N- (4-fluoro-3- (trifluoromet¡Dphenyl) -l- (2-oxo-2 - (((R) -l, l, ltrifluoropropan-2-inamino) acetyl ) pyrrolidin-3-carboxamide

<img file="MX2016005342A_D0035.tif" />

Compound 14 was prepared according to the method described for compound 1, with the exception that, in step 1, 4-fluoro-3- (trifluoromethyl) aniline was used instead of 3-bromo-4, 5difluoroaniline. The coupling reaction to give the title compound was performed according to the procedure described for compound 13, step two, with the exception that (/?) L, l, l-trifluoro-2-propylamine was used instead of l - (trifluoromethyl) -cyclopropanamine. Method B, Rt - 1.01 min, m / z = 442.1 (MH), exact mass: 443.1. ^TH NMR (400 MHz, DMSO - &lt;/₆) δ ppm 1.30 (d, J = 7.Q

Hz, 3 Η), 1.87 - 2.37 (m, 2 Η), 3.13 - 3.27 (m, 1 Η), 3.37 - 3.98 (m, 4 Η), 4.34 - 4.77 (m, 1 Η), 7.41

- 7.55 (m, 1 Η), 7.76 - 7.90 (m, 1 Η), 8.01 - 8.25 (m, 1 Η), 9.27 (bs, 1 H), 10.50 (bs, 1 H)

Compound 15: (S) -N- (3-chloro-4-fluorophen¡nl- (2-oxo-2 - (((R) -l, l, l-trifluoropropan2-yl) amino) acetyl) p¡rrolid¡n-3-carboxam¡da

<img file="MX2016005342A_D0036.tif" />

Compound 15 was prepared according to the methods described for the synthesis of compound 1, with the exception that, in step one, 3-chloro-4-fluoroaniline was used instead of 3bromo-4,5-difluoroaniline. The coupling reaction to give the title compound was performed according to the procedure described for compound 13, step two, with the exception that (/?) - l, l, l-trifluoro-2-propylamine was used instead of l- (trifluoromethyl) -cyclopropanamine. Method B, Rt 0.96 min, m / z = 408.1 (MH) ', exact mass: 409.1. * H NMR (400 MHz, DMSO-z £) δ ppm 1.30 (d, J = 7.0 Hz, 3 H), 1.91 - 2.30 (m, 2 H), 3.10 - 3.27 (m, 1 H), 3.38 - 4.02 (m, 4 H), 4.52 - 4.71 (m, 1 H), 7.32 - 7.41 (m, 1 H), 7.43 - 7.51 (m, 1 H), 7.86 - 7.99 (m, 1 H), 9.26 (sa , 1H), 10.34 (bs, 1H), a mixture of rotamers.

Compound 16: (S) -N- (3-chloro-4,5-difluorophenyl) -l- (2-oxo-2 - ((l, l, l-tr, fluoro-2-methylDroDan-2-yl) amino) acet¡nD¡rrol¡d¡n-3-carboxam¡da

<img file="MX2016005342A_D0037.tif" />

Compound 16 was prepared according to the method of preparing compound 13 with the exception that l, l, l-trifluoro-2-methylpropan-2-amine was used in step two, instead of 1 (trifluoromethyl) - dclopropanamine. Method B, Rt = 1.08 min, m / z = 440.1 (MH)^-, exact mass: 441.1. ^JH NMR (400 MHz, DMSO-í /₆) δ ppm 1.54 (s, 6H) 1.98 - 2.31 (m, 2H) 3.06 - 3.28 (m, 1H) 3.40 - 3.97 (m, 4H) 7.50 - 7.80 (m, 2H) 8.56 (m, 1H) 10.44 (bs, 1H), as a mixture of rotamers.

Synthesis of Compound 17 N ^ -fluoro-S-methylphenyl-S-metlI-l-fZ-fS-metlIoxetan-Sil) amino) -2-oxoacetyl) prolrolid-3-carboxamide.

<img file="MX2016005342A_D0038.tif" />

Step 1. Preparation of l- (r-butoxycarbonyl) -5-methylprolopin-3-carboxylic acid. The title compound was prepared as a mixture of diastereomers according to methods found in W02010059658 (p. 211), starting from methyl 2-chloro-5-methyl-lH-pyrrole-3-carboxylate described in Foley, L ., Tetrahedron Letters 1994, vol. 35, p. 5989.

Step 2. Preparation of f-Butyl 4-((4-fluoro-3-methylfenyl) carbamoiD-2-methylpyrrolidin-carboxylate, 4-Fluoro-3-methylaniline (1.09 g, 8.72 mmol) to a solution of l- (f-butoxycarbonyl) -5-methylprolopin-3-carboxylic acid (2 g, 8.72 mmol), DIPEA (4.33 ml, 26.17 mmol), and HATU (4.98 g, 14.09 mmol) in CH₂CI₂ (50 mi). The reaction mixture was stirred for 1 h at room temperature, then partitioned with water. The organic phase was removed, dried over MgSO₄Solids were removed by filtration and the solvent was removed from the filtrate under reduced pressure. The crude was purified by silica gel column chromatography resulting in the title compound. Method C, Rt = 1.96 min, m / z = 335.0 (MH) ', and 1.98 min, m / z - 335.1 (MH)' exact mass: 336.2.

Step 3. Ethyl ethyl 2- (4 - ((4-fluoro-3-methylphenyl) carbamo, D-2-methylp, rrol, dl-l) 2-oxoacetate. TFA was added dropwise to a solution of i-butyl 4-((4-0uoro-3-methylphenyl) carbamoyl) -2-methylpyrrolidin-l-carboxylate in CH₂CI₂ under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure and the crude was reconstituted in CH₂CI₂ and NaOH (1M, aq.). The mixture was vigorously stirred for 5 minutes. The phases were separated and the aqueous phase was extracted with CH₂CI₂. The combined organic phases were dried over MgSO₄Solids were removed by filtration and the filtrate was concentrated under reduced pressure to give an oil. CH were added₂CI₂anhydrous (50 ml) and triethylamine (1.09 g, 7.83 mmol) to this oil. Ethyloxalyl chloride (0.44 ml, 3.92 mmol) was added dropwise to the resulting solution at room temperature, then stirred for 18 hours. HCI (0.5 M aq.) Was added to the reaction mixture. The organic phase was removed, dried over MgSO₄The solids were removed by filtration and the filtrate was concentrated to give an oil, dried in vacuo at 50 ° C for 4 hours and used without further purification.

Step 4. Preparation of 2- (4 - ((4-fluoro-3-methylphenyl) carbamoyl) -2-methylpyrrolyldln-iD-2-oxoacetic acid. The ester hydrolysis of 2 Ethyl ethyl (4 - ((4-fluoro-3-methylphenyl) carbamoyl) -2-methylpyrrolydin-lyl) -2-oxoacetate was achieved according to the method described in step 7 of compound 10.

Step 5. Preparation of / V- (4-fluoro-3-methylphenyl) -5-methyll- (2 - ((3-methylhexeta-3yl) amino) -2-oxoacetyl) pyrrolidin-3- Carboxamide The title compound was prepared according to the procedure in step 8 in the synthesis of compound 10. Isomers were isolated by preparative SFC (stationary phase: Whelk-0 (R, R) 20 x 250 mm, mobile phase: CO₂, EtOH / iPrOH (50/50) with 0.2% ¡PrNH₂). The desired fractions were collected and the solvent was removed under reduced pressure to give compounds 17a (119 mg), 17b (116 mg), 17c (78 mg), and 17d (94 mg) named in order of elution.

<td>Compound</td><td>LC-MS Method, Rt (min)</td><td>m / z (M + H)⁺</td><td>Setting</td>

<td>17a</td><td>C, 1.39</td><td> 378.2</td><td>(3R, 5S) 0 (3S, 5R)</td>

<td>17b</td><td>C, 1.39</td><td> 378.2</td><td>(3R, 5S) or (3S, 5R)</td>

<td>17c</td><td>C, 1.37</td><td> 378.2</td><td>(3S, 5S) or (3R, 5R)</td>

<td>17d</td><td>C, 1.37</td><td> 378.2</td><td>(3S, 5S) 0 (3R, 5R)</td>

Compound 17a: NMR (600 MHz, DMSO-í /₆) δ ppm 1.21 (d, 7 = 6.3 Hz, 3 H), 1.26 (d, J = 6.2 Hz, 3 H), 1.53 (s, 3 H), 1.54 (s, 3 H), 1.75 (ddd, 7 = 12.7, 10.1, 8.1 Hz, 1 H), 1.87 (ddd, 7 = 13.0, 7.5, 5.6 Hz, 1 H), 2.19 - 2.22 (m, 6 H), 2.41 (dt, J = 12.6, 7.5 Hz, 1H), 2.46 - 2.53 (m, 1H), 3.01 - 3.12 (m, 2H), 3.52 (dd, 7 = 12.2, 7.9 Hz, 1H), 3.65 (dd, 7 = 11.4, 9.8 Hz, 1 H), 3.90 (dd, 7 = 12.2, 8.1 Hz, 1 H), 4.01 - 4.07 (m, 1 H), 4.09 (dd, 7 = 11.4, 7.5 Hz, 1 H), 4.29 - 4.35 (m, 4H), 4.37 - 4.48 (m, 1H), 4.62 - 4.67 (m, 4H), 7.05 - 7.09 (m, 2H), 7.37 - 7.42 (m, 2H), 7.49 - 7.53 (m, 2H), 9.19 (s, 1H), 9.23 (s, 1H), 10.02 (s, 1H), 10.04 (s, 1H) ), as a mixture of rotamers.

Compound 17b: ^JH NMR (400 MHz, DMSO-cfc) δ ppm 1.21 (d, 7 = 6.2 Hz, 3 H), 1.26 (d, Jt = 6.2 Hz, 3 H), 1.49 - 1.56 (m, 6 H), 1.75 ( ddd, 7 = 12.7, 10.0, 8.0 Hz, 1 H), 1.87 (ddd, 7 = 13.0, 7.4, 5.8 Hz, 1 H), 2.17 - 2.23 (m, 6 H), 2.41 (dt, 7 = 12.7, 7.5 Hz, 1 H), 2.45 - 2.54 (m, 1 H), 2.96 3.13 (m, 2 H), 3.52 (dd, 7 = 12.1, 7.9 Hz, 1 H), 3.65 (dd, 7 = 11.4, 9.8 Hz, 1 H), 3.91 (dd, 7 = 12.2, 8.0 Hz, 1 H), 3.98 - 4.15 (m, 2 H), 4.27 - 4.36 (m, 4 H), 4.37 - 4.49 (m, 1 H) , 4.59 - 4.70 (m, 4H), 7.07 (t, 7 = 9.1Hz, 2H), 7.34 - 7.44 (m, 2H), 7.46 - 7.55 (m, 2H), 9.18 (s, 1H), 9.22 (s, 1H), 10.01 (s, 1H), 10.03 (bs, 1H ), as a mixture of rotamers.

Compound 17c: ^LH NMR (400 MHz, DMSO-cQ δ ppm 1.13 - 1.27 (m, 6 H), 1.51 (s, 3 H), 1.53 (s, 3 H), 1.86 (ddd, 7 = 12.3, 6.8, 2.9 Hz, 1 H), 1.98 (dd, 7 = 12.0, 6.9 Hz, 1 H), 2.07 - 2.17 (m, 2 Η), 2.18 - 2.23 (m, 6 Η), 3.26 - 3.31 (m, 2 Η), 3.58 - 3.70 (m, 2 Η), 3.84 (dd, 7 = 11.7, 7.9 Hz, 1H), 3.92 - 4.01 (m, 1H), 4.17 - 4.26 (m, 1H), 4.27 - 4.36 (m, 4H), 4.54 - 4.62 (m, 1H), 4.61 4.66 (m, 4H), 7.01 - 7.12 (m, 2H), 7.32 - 7.43 (m, 2H), 7.47 - 7.57 (m, 2 H), 9.17 (s, 1H), 9.20 (s, 1H), 10.03 (s, 1H), 10.07 (s, 1H), as a mixture of rotamers.

Compound 17d: * H NMR (600 MHz, DMSO-ofe) δ ppm 1.20 (d, 7 = 6.5 Hz, 3 H), 1.21 (d, 7 = 6.5 Hz, 3 H), 1.51 (s, 3 H), 1.53 (s , 3 H), 1.86 (ddd, 7 = 12.3, 6.8, 2.9 Hz, 1 H), 1.98 (dd, 7 = 12.1, 6.8 Hz, 1 H), 2.10 - 2.18 (m, 2 H), 2.18 - 2.23 (m, 6H), 3.28 - 3.32 (m, 2H), 3.60 - 3.68 (m, 2H), 3.84 (dd, 7 = 11.6, 7.9 Hz, 1H), 3.97 (dd, 7 = 11.7, 7.8 Hz, 1 H), 4.18 - 4.26 (m, 1 H), 4.28 4.35 (m, 4 H), 4.56 - 4.61 (m, 1 H), 4.62 - 4.67 (m, 4 H), 7.03 - 7.11 ( m, 2 H), 7.35 - 7.42 (m, 2 H), 7.48 - 7.55 (m, 2 H), 9.19 (s, one H), 9.22 (s, 1H), 10.04 (s, 1H), 10.09 (s, 1H), as a mixture of rotamers.

Compound 18: Μ (3- &lt;ΙθΓθ-4.5 ^ ίΑυθΓθί6ηιΌ-2.2 ^ ίηΊ6ϋΙ-1-Γ2-οχο-2-ΓΓ (1 /?) - 2.2,2trifluoro-l-methylethylaminolacetylpyrrolidine-3-carboxamide

F

<img file="MX2016005342A_D0039.tif" />

A mixture of diethyl fumarate (19.05 ml / 113,848 mmol) and 2-nitropropane (10.2 ml / 113.8 mmol) was treated with KF / basic alumina (20 g). The reaction mixture was stirred overnight and the mixture was filtered. The filtrate was concentrated to provide crude diethyl 2- (l-methyl-l-nitroethyl) butanedioate (20 g) which was used as such.

^JH NMR (400 MHz, DMSO-d₆) δ ppm 1.10 - 1.22 (m, 6H) 1.54 (s, 3H) 1.58 (s, 3H) 2.55 - 2.76 (m, 2H) 3.52 (dd, J = 11.00, 3.96 Hz, 1H) 3.99 - 4.13 (m, 4H). Pd / C (10%) (448.04 .mg / 0.421 mmol) was added to a solution of crude Diethyl 2- (l-methyl-l-nitro-ethyl) butanedioate (2200 mg, 8.42 mmol), triethylamine (1.17 ml / 8.42 mmol) and ethanol (100 ml) under a flow of nitrogen. The resulting mixture was stirred under a hydrogen atmosphere at room temperature until 3 equivalents of hydrogen were absorbed. The catalyst was removed by filtration over decalite and the filtrate was evaporated to produce ethyl 2,2-dimethyl-5-oxo-pyrrolidin-3-carboxylate (1.05 g) as a solid which was used as such. A mixture of ethyl 2,2-dimethyl-5-oxo-prolidin-3-carboxylate (750 mg / 4.05 mmol) and Lawesson's reagent (983 mg / 2.43 mmol) in toluene was heated on molecular sieves (15 ml ) to 70 ° C for 1 hour, cooled and concentrated in vacuo, resulting in a solid residue. The crude was purified using silica gel column chromatography (gradient elution: EtOAc-heptane, 0: 100 to 100: 0) to give ethyl 2,2-dimethyl-5-thioxo pyrrolidin-3-carboxylate (432 mg ) as a slightly yellow powder, which was used as such. Method B, Rt = 0.66 min, m / z = 202.1 (M + H)⁺, exact mass: 201.1. Ethyl 2,2-dimethyl-5-thioxopyrrolidin-3-carboxylate (100 mg, 0.5 mmol) was dissolved in tetrahydrofuran (2 ml). Ethanol (2 ml) was added thereto and the mixture was stirred overnight. The mixture was filtered over a bed of decalite, rinsed with ethanol and concentrated in vacuo to provide crude ethyl 2,2-dimethylpyrroletin-3-carboxylate (50 mg) as a beige powder which was used as such .

Ethyloxalyl chloride (65.35 μΙ / 0.58 mmol) was added dropwise to a solution of crude ethyl 2,2-dimethylpyrrole-3-carboxylate (50 mg, 0.29 mmol) and DIPEA (0.25 ml / 1.46 mmol ) in CH₂CI₂ (2 ml) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. NaHCO added₃ saturated aqueous (5 ml) and CH₂CI₂ (5 ml) to the reaction mixture and the phases were separated. The organic phase was dried over MgSO₄, filtered and evaporated to dryness. The obtained residue was purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100). The desired fractions were concentrated in vacuo to provide ethyl l- (2-ethoxy-2-oxo-acetyl) -2,2-dimethyl-pyrrolidin-3carboxylate (80 mg) as a clear, colorless oil that was used as such. Ethyl 1 (2-ethoxy-2-oxo-acetyl) -2,2-dimethyl-pyrrolidin-3-carboxylate (80 mg, 0.29 mmol) was dissolved in ethanol (1 ml / 17.13 mmol) and cooled in a bath of ice. NaOH (0.59 ml / 1M / 0.59 mmol) was added and the mixture was stirred while continuing to cool for 10 minutes. HCI (0.59 ml, 1M, 0.59 mmol) was added dropwise with cooling. The mixture was concentrated in vacuo. The residue was partitioned between water and Me-THF. The organic phase was separated, dried (Na₂SW₄), filtered and concentrated in vacuo, resulting in 2- (3-ethoxycarbonyl-2,2-dimethyl-pyrrolidin-l-yl) -2-oxo-acetic acid (70 mg) as an oil that was used as such. A solution of Z-IS-ethoxycarbonyl ^^ - dimethylpyrrolidin-l-yl) -2-oxoacetic acid (70mg, 0.29mmol) in DMF (10ml) was cooled to 5 ° C in an ice-water bath. DIPEA (0.15 ml, 0.75 g / ml, 0.86 mmol) and (R) -l, l, l-trifluoro-2-propylamine (39.05 mg, 0.35 mmol) were then added and stirred. A solution of HATU (120.36 mg, 0.32 mmol) in DMF (5 ml) was added dropwise while continuing to cool. The obtained solution was stirred for 1 hour with cooling. The reaction was quenched with water and neutralized with a 1N HCI solution. Brine (10 ml) was added and the compound was extracted with EtOAc (3 x 20 ml). The combined organic phases were dried with Na₂SW₄, filtered and evaporated to dryness. This was purified by flash column chromatography on silica, from heptane to EtOAc (100/0 - 0/100). The desired fractions were collected and evaporated to dryness to obtain 2,2-dimethyll- [2-oxo-2 - [[(lR) -2,2,2-trifluoro-l-methyl-ethyl] am Ethyl] acetyl] pyrrolidin-3-carboxylate (70 mg) as a white solid which was used as such. 2,2-Dimethyl-l- [2-oxo-2 [[(lR) -2,2,2-trifluoro-l-methyl-ethyl] amino] acetyl] pyrroxydin-3- was dissolved. Ethyl carboxylate (70mg, 0.21mmol) in THF (5ml). LiOH was added thereto (17.7 mg, 0.74 mmol) in water (5 ml). MeOH (0.2 mL) was added to dissolve all reagents. The mixture was stirred overnight at room temperature. Then it was concentrated in vacuo until only water remained. Next, HCI (0.74 ml, 1M, 0.74 mmol) was added and extracted using Me-THF (3 X 10 ml). The combined extracts were washed with brine (20 ml), dried over Na₂SW₄, filtered and concentrated in vacuo to obtain 2,2-dimethyll- [2-oxo-2 - [[(l /?) - 2,2,2-trifluoro-l-methyl methyl acid ] amino] acetyl] pyrrolidin-3-carboxylic acid (45 mg) as a white powder that was used as such. 2,2-Dimethyl-l- [2-oxo-2 [[(l /?) - 2,2,2-trifluoro-l-methylethyl] amino] acet¡l] p¡rrol¡d¡ was dissolved. n-3-carboxylic (45mg, 0.15mmol), 3-chloro-4,5difluoro-aniline (58.02mg, 0.29mmol), HATU (110.3mg, 0.29mmol) and DIPEA (0.12ml, 0.75g / ml , 0.73 mmol) in DMF (0.34 ml, 4.34 mmol). This mixture was stirred at room temperature for 2 hours. Extra DIPEA (0.12 ml, 0.75 g / ml, 0.73 mmol) was added and the mixture was stirred at 60 ° C for 2 hours. This mixture was purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100) and further by preparative HPLC (stationary phase: Uptisphere C18 ODB-10 pm, 200g, 5 cm, mobile phase: NH dissolution₄HCO₃ 0.25% in water, MeOH). The desired fractions were concentrated in vacuo, coevaporated twice using MeOH and dried in a vacuum oven at 55 ° C for 24 hours to provide N- (3-chloro-4,5-difluoro-phenyl) -2,2- d-methyl-l- [2-oxo-2 - [[(lR) -2,2,2-trifluoro-l-methylethyl] amino] acetyl] pyrroyl-d-3-carboxamide (6.3 mg) as a white solid. NMR (400 MHz, CHLOROFORM-d) δ ppm 1.35 - 1.39 (m, 3H), 1.46 - 1.49 (m, 3H), 1.69 - 1.80 (m, 3H), 2.01 - 2.20 (m, 1 H), 2.23 - 2.43 (m, 1H), 2.58 - 2.74 (m, 1H), 3.86 - 4.09 (m, 1H), 4.20 - 4.47 (m, 1H), 4.48 - 4.67 (m, 1 H), 7.08 (s, 1H), 7.28 - 7.36 (m, 1H), 7.41 - 7.49 (m, 1H), 7.49 - 7.65 (m, 1H). LC method B; Rt: 1.11 min. m / z: 454.2 (MH) - Exact mass: 455.1

Compound 19: (3S) -l-r2- (tert-butylamine) -2-oxo-acetyl1-N- (3-chloro-2,4-difluorophenyljpyrrolidin-3-carboxamide ci

<img file="MX2016005342A_D0040.tif" />

Ethyl 2 - [(3S) -3 - [(3-chloro-2,4-difluoro-phenyl) carbamoyl] pyrrolin-l-yl] -2-oxoacetate was obtained in a similar manner as described for Ethyl (S) -2- (3 - ((3-bromo-4,5-difluorophenyl) carbamoyl) pyrrolidin-l-yl) -2-oxoacetate using 3-chloro-2,4-difluoro-aniline instead of 3-bromo-4,5difluoroaniline in step one. Ethyl 2 - [(3S) -3 - [(3-chloro-2,4-difluoro-phenyl) carbamoyl] pyrrolidin-yl] -2-oxo-acetate (0.6 g, 1.66 mmol) was dissolved in tetrahydrofuran (15 me). To this was added tert-butylamine (0.18 g, 2.49 mmol) and the mixture was cooled in an ice-water bath. Lithium bis (trimethylsilyl) amide (1M in toluene) (4.99 ml, 1M, 4.99 mmol) was then added dropwise over a period of 5 minutes. The resulting mixture was stirred for 1 hour while cooling continued. Then it went extinct using NH₄CI (saturated / 50 mi). This was extracted using EtOAc (3 x 50 ml). The combined extracts were washed with brine (50 ml), dried with Na₂SW₄, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100) and further by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD-10 pm, 30x150 mm, mobile phase: NH dissolution₄HCO₃ 0.25% in water, MeOH), to provide compound 19 (136 mg) as a white powder. Method B, Rt = 0.95 min, m / z = 386.2 (MH) ', Exact Mass: 387.1. NMR (400 MHz, DMSO-d₆) δ ppm 1.31 (s, 9H), 1.85 - 2.30 (m, 2H), 3.15 - 4.33 (m, 5H), 7.26 - 7.34 (m, 1H), 7.65 - 7.86 (m, 1H) , 8.00 (m, 1H), 10.08 (bs, 1H) as a mixture of rotamers.

Compound ________ 20: ________ (3S ~) -l-r2- (tert-butylamine) -2-oxo-acetyl1-N- (3-cyano-4fluorophenyl) prrolidin-3-carboxamide

<img file="MX2016005342A_D0041.tif" />

Compound 20 was prepared in a similar manner to that described for compound 19, using 5-amine-2-difluorobenzonyl in place of 3-bromo-2,4-fluoroaniline in step one. Method D, Rt = 1.66 min, m / z = 359.1 (MH) ', Exact mass: 360.2. * H NMR (400 MHz, DMSO-d₆) δ ppm 1.30 (m, 9H), 1.92 - 2.29 (m, 2H), 3.06 - 3.27 (m, 1H), 3.34 - 4.01 (m, 4H), 7.38 - 7.58 (m, 1H) , 7.77 - 7.89 (m, 1H), 7.91 - 8.07 (m, 1H), 8.09 - 8.19 (m, 1H), 10.32 - 10.59 (m, 1H) as a mixture of rotamers.

Compound 21: (3S) - / V- (3-chloro-2,4-dfluorophenyl) -l-12-oxo-2-rr (l /?) - 2,2,2-trfluoro -lmethyl-ethylaminolacetylpyrrolidin-3-carboxamide

<img file="MX2016005342A_D0042.tif" />

<img file="MX2016005342A_D0043.tif" />

Compound 21 was prepared in a similar manner to that described for compound 19 using (R) -l, l, l-trifluoro-2-propylamine instead of tert-butylamine. Method B, Rt = 0.97 min, m / z = 426.2 (MH) ', Exact Mass: 427.1.^XH NMR (400 MHz, DMSO-d₆) δ ppm 1.27 - 1.33 (m, 3H), 1.95 - 2.28 (m, 2H), 3.33 - 4.00 (m, 5H), 4.52-4.72 (m, 1H), 6.97 - 7.48 (m, 1 H), 7.60 - 7.91 (m, 1H), 9.01 - 9.47 (m, 1H), 9.90 - 10.28 (m, 1H) as a mixture of rotamers.

Compound 22: (3S) - / V- (3-cyano-4-fluorophenyl ') - l-r2-oxo-2-rail /?) - 2,2.2-trifluoro-l-methyl-ethylaminolacetylpyrrolidin-3-carboxamide

<img file="MX2016005342A_D0044.tif" />

Compound 22 was prepared in a similar manner to that described for compound 20 using (R) -l, l, l-trifluoro-2-propylamine instead of tert-butylamine. Method B, Rt = 0.87 min, m / z = 399.2 (MH) ', Exact Mass: 400.1.^JH NMR (400 MHz, DMSO-d₅) δ ppm 1.30 (d,&gt; 7.0 Hz, 3H), 1.96 - 2.30 (m, 2H), 3.11 - 3.28 (m, 1H), 3.38 - 4.00 (m, 4H), 4.41 - 4.77 (m , 1 H), 7.42 - 7.56 (m, 1 H), 7.78 - 7.90 (m, 1 H), 8.04 - 8.23 ​​(m, 1 H), 9.26 (bs, 1H), 10.50 (bs, 1H) like a mixture of rotamers.

Compound 23: (3S) -N-r4-fluoro-3- (trifluoromet¡Dfen¡l1-l-r2- (¡sopropilamino) -2-oxoacetillpirrolidin-3-carboxamide

<img file="MX2016005342A_D0045.tif" />

Compound 23 was prepared in a similar manner to that described for compound 14 using isopropylamine instead of (R) -l, l, l-trifluoro-2-propylamine. Method B, Rt = 0.94 min, m / z = 388.2 (MH), Exact mass: 389.1.^XH NMR (400 MHz, DMSO-d₆) δ ppm 1.00 - 1.17 (m, 6H), 1.94 - 2.30 (m, 2H), 3.10 - 3.26 (m, 1H), 3.35 - 4.02 (m, 5H), 7.36 - 7.58 (m, 1 H), 7.75 - 7.95 (m, 1H), 8.04 - 8.19 (m, 1H), 8.36 - 8.53 (m, 1H), 10.37 10.63 (m, 1H) as a mixture of rotamers.

Compound ________ 24: ________ (3S) -N-r4-fluoro-3- (trifluoromethyl) fen¡l1-l-r2-rr (lR) -lmet¡lprop¡l1am¡no1-2-oxo-acetyl1p¡rrolid¡n -3-carboxam¡da

Compound 24 was prepared in a similar manner to that described for the compound using (R) - (-) - 2-aminobutane instead of (R) -l, l, l-trifluoro-2-propylamine. Method B, Rt = 0.99 min, m / z = 402.2 (MH) ', Exact Mass: 403.2 / 14 NMR (400 MHz, DMSO-d₆) δ ppm 0.76 - 0.88 (m, 3H), 1.00 - 1.15 (m, 3H), 1.35 - 1.53 (m, 2H), 1.94 - 2.29 (m, 2H), 3.11 - 3.26 (m, 1 H), 3.37 - 4.01 (m, 5H), 7.40 - 7.53 (m, 1H), 7.79 - 7.89 (m, 1H), 8.05 - 8.16 (m, 1H), 8.29 - 8.46 (m, 1 H), 10.35-10.60 (m, 1H) as a mixture of rotamers.

Compound 25: (3S) -N- (3-chloro-4-fluorophenyl) -l-r2-oxo-2-rr 1-trifluoromethyl) cidopropinaminolacetiDpirrolidin-S-carboxamide

Cl

Compound 25 was prepared in a similar manner to that described for the compound using l- (trifluoromethyl) cyclopropan-l-amine instead of (R) -l, l, l-trifluoro-2-propylamine.

Method B, Rt = 0.97 min, m / z = 420.1 (MH) ', Exact Mass: 421.1. ^ NMR (400

MHz, DMSO-d₆) δ ppm 0.95 - 1.14 (m, 2H), 1.22 - 1.29 (m, 2H), 1.95 - 2.29 (m, 2H), 3.09 - 3.24 (m, 1H), 3.34 - 3.98 (m, 4 H), 7.32 - 7.41 (m, 1 H), 7.42 - 7.53 (m, 1 H), 7.88 - 7.97 (m, .1 H), 9.44 (s, 1 H), 10.19 - 10.35 (m, 1 H ) as a mixture of rotamers.

Compound 26: (3S) - / V- (3-chloro-4-fluorophenyl) -l-r2-oxo-2-rr (lS) -2,2,2-trifluoro-l-methyl-ethylaminolacetylpyrrolidin-3-carboxamide

<img file="MX2016005342A_D0046.tif" />

Compound 26 was prepared in a similar manner to that described for compound 15 using (S) -l, l, l-trifluoro-2-propylamine instead of (R) -l, l, l-trifluoro-2-propylamine . Method B, Rt = 0.97 min, m / z = 408.1 (MH) ', Exact Mass: 409.1^LH NMR (400 MHz, DMSO-d₆) δ ppm 1.26 - 1.37 (m, 3H), 1.95 - 2.29 (m, 2H), 3.10 - 3.27 (m, 1H), 3.34 - 3.98 (m, 4H), 4.52 - 4.71 (m, 1 H), 7.32 - 7.41 (m, 1 H), 7.43 - 7.52 (m, 1 H), 7.86 - 7.99 (m, 1 H), 9.17 - 9.33 (m, 1 H), 10.22 10.35 (m, 1 H) ) as a mixture of rotamers

Compound 27: (2S) - / V- (3-cyano-4-fluorophenyl) -lT2- (isopropylamine) -2-oxoacetyl1-2methyl-pyrrolidin-3-carboxamide

<img file="MX2016005342A_D0047.tif" />

Methyl (2S, 3S) -2-methyl-1 - ((S) -l-phenylethyl) pyrroletin-3-carboxylate (1.9 g, 7.68 mmol) was dissolved in methanol (50 ml). This was added to Pd / C (10% / 0.82 g, 0.77 mmol) under nitrogen. The mixture was stirred under a hydrogen atmosphere at room temperature for 24 hours. The resulting mixture was filtered over a bed of decalite and rinsed using methanol (100 ml). The filtrate was concentrated in vacuo to provide methyl (2S, 3S) -2-methylpyrrolidin-3-carboxylate (830 mg) as a clear oil. 2-Chloro-2-oxo-ethyl acetate (1.3 ml, 11.59 mmol) was added dropwise to a solution of methyl (2S, 3S) -2-methylpyrrolidin-3-carboxylate (0.83 g, 5.8 mmol) and diisopropylethylamine (4.99 ml, 28.98 mmol) in dry dichloromethane (5 ml) at room temperature. The reaction mixture was stirred at room temperature for 1 hr.

NaHCO added₃ saturated aqueous (5 ml) to the reaction mixture and the phases were separated. It was then extracted using dichloromethane (2 x 10 ml). The combined extracts were dried over Na₂SW₄, filtered and concentrated in vacuo. The crude obtained was purified by means of silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100). The desired fractions were concentrated in vacuo to provide methyl (2S, 3S) -l- (2-ethoxy-2-oxo-acetyl) -2-methyl, lpr, rdn-3-carboxylate (890 mg) as a yellow oil.

Methyl (2S, 3S) -l- (2-ethoxy-2-oxo-acetyl) -2-methyl-pyrrole-3-carboxylate (250 mg, 1 mmol) was dissolved in ethanol (10 ml) and isopropylamine (1698 pl, 19.94 mmol) and the mixture was stirred at 60 ° C for 2 hours. The mixture was concentrated in vacuo. The obtained oil was purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100). The desired fractions were concentrated under reduced pressure to provide (2S) -l- [2- (isopropylamino) -2-oxo-acetyl] -2-metylprrrolidin-3-carboxylate of methyl (380 g) as a clear oil that was used as such.

Methyl (2S) -l- [2- (isopropylamine) -2-oxo-acetyl] -2-methlp, rrol, dn-3-carboxylate (0.38 g, 1.48) was dissolved mmol) in tetrahydrofuran (10 ml) and this was stirred at room temperature. To this was added LiOH (178 mg, 7.41 mmol) in water (2 ml) followed by methanol (2 ml). The resulting mixture was stirred at room temperature for 2 hours. Then, HCI (1 M in H₂O) (7.41 ml, 1M, 7.41 mmol) and the mixture was concentrated in vacuo until only water remained. Water (5 ml) was added and this solution was extracted using 2-methyl-tetrahydrofuran (3 x 15 ml). The combined extracts were washed with brine (15 ml), dried over Na₂SW₄, filtered and concentrated in vacuo to obtain (2S) -l- [2- (isopropylamino) -2-oxo-acetyl] -2-methyl-pyrrolidin-3-carboxylic acid (312 mg) which was used as such.

(2S) -l- [2- (isopylamino) -2-oxo-acetyl] -2-methyl-pyrroletin-3-carboxylic acid (104 g, 0.43 mmol) was dissolved in Ν, Ν-dimethylformamide (1 me). HATU (0.18 g, 0.47 mmol) was then added and this mixture was stirred for 20 minutes. DIPEA (0.22 ml, 0.75 g / ml, 1.29 mmol) was then added followed by 5-amino-2-fluorobenzonitrile (0.12 mg, 0.86 mmol). The reaction mixture was stirred at 50 ° C for 4 hours. This mixture was then cooled to room temperature and injected directly onto a bed of silica. The mixture was purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100) and further by preparative HPLC (stationary phase: RP SunFire Prep C18 OBD-10 pm, 30x150 mm; mobile phase: NH dissolution₄HCO₃ 0.25% in water, MeOH). The desired fractions were concentrated under reduced pressure and coevaporated twice with methanol (2 X 15 ml) and dried in a vacuum oven at 55 ° C for 18 hours to provide compound 27 (57 mg) as a white powder. Method B, Rt = 0.81 (31%) and 0.83 min (69%), m / z = 359.2 (MH) ', Exact mass: 360.2

Compound 28: (2S) -M (3-chloro-2,4-dfluoro-phenyl) -l-r2- (isopropylamine ') - 2-oxoacetyll1-2-methlp¡rrol¡din-3 -carboxam¡da

<img file="MX2016005342A_D0048.tif" />

Compound 28 was prepared from (2S) -l- [2- (isopropylamino) -2-oxoacetyl] -2-methyl-pyrrolidin-3-carboxylic acid in a similar manner as described for compound 27 , using 3-chloro-2,4-difluoro-aniline instead of 5-amino-2-fluorobenzonitrile. Method B, Rt = 0.91 (48%) and 0.92 min (52%), m / z - 386 (MH) ', Exact mass: 387.1.

Compound 29: (2S ') - / y- (3-chloro-4,5-dfluoro-phenyl) -l-r2- (isopropylamino) -2-oxoacetill ^ -methyl-pyrrolidin-S-carboxamide

<img file="MX2016005342A_D0049.tif" />

Compound 29 was prepared from (2S) -l- [2- (isopropylamino) -2-oxoacetyl] -2-methyl-pyrrolidin-3-carboxylic acid in a similar manner as described for compound 27, using 3- chloro-4,5-difluoro-aniline instead of 5-amino-2-fluorobenzonitrile. Diastereomeric mixture 29 (63 mg) was separated by preparative SFC (stationary phase: Chiralpak Diacel AD 20 x 250 mm; mobile phase: CO₂, MeOH with 0.2% of PrNH₂), resulting in 29a (second eluent, 20 mg) and 29b (first eluent, 13.2 mg after further purification by silica gel column chromatography using a gradient elution from heptane to l PrOH (from 100: 0 to 65: 35)). 29: Method B, 0.98 (42%) and 1.02 min (58%), m / z = 386 (MH) ', Exact mass: 387.1.29a: Method D, Rt = 1.89, m / z = 386.1 (MH) ', Exact mass: 387.1; NMR (400 MHz, DMSO-d₆) δ ppm 0.95 - 1.05 (m, 3H), 1.06 - 1.16 (m, 6H), 1.82 - 2.11 (m, 1H), 2.14 - 2.44 (m, 1H), 3.04 - 3.26 (m, 1 H), 3.35 - 4.10 (m, 3H), 4.32 - 4.97 (m, 1H), 7.33 - 7.85 (m, 2H), 8.20 - 8.73 (m, 1H), 10.07 - 10.68 (m, 1 H) as a mixture of rotamers. 29b: Method B, Rt = 0.97 m / z = 386.2 (ΜΗ) ', Exact mass: 387.1. 'H NMR (400 MHz, DMSO-d₆) δ ppm 1.03 -1.14 (m, 6H), 1.23 - 1.31 (m, 3H), 1.93 - 2.11 (m, 1H), 2.14 - 2.30 (m, 1H), 2.72 - 2.93 (m, 1 H), 3.30-4.70 (m, 4H), 7.56 - 7.73 (m, 2H), 8.28 - 8.54 (m, 1H), 10.22-10.60 (m, 1H) as a mixture of rotamers.

Compound 30: (3S) - / V-r3-dfluoromethyl-4-fluorophenyl-1-l-r2-oxo-2-rí (lS) -2,2,2trifluoro-l-methyl-ethylaminoJacetillpyrrolidin-3-carboxamide

<img file="MX2016005342A_D0050.tif" />

<img file="MX2016005342A_D0051.tif" />

<img file="MX2016005342A_D0052.tif" />

Ethyl 2 - [(3S) -3 - [[3- (difluoromethyl) -4-fluoro-phenyl) carbamoyl] pyrrolidin-l-yl] -2-oxoacetate was obtained in a similar way as described for (S) - Ethyl 2- (3 - ((3-bromo-4,5difluorophenyl) carbamoyl) pyrroyl-l-l) -2-oxoacetate using 3- (difluorometrl) -4-fluoro-aniline instead of 3-bromo -4,5-difluoroaniline. Compound 30 was prepared from ethyl 2 - [(3S) -3 - [[3 (difluoromethyl) -4-fluoro-phenyl] carbamoyl] pyrrolidin-l-yl] -2-oxo-acetate in a manner similar to that described for the synthesis of compound 19 from 2 - [(3S) -3 - [(3-chloro-2,4-difluorophenyl) carbamoyl] pyrrolidyl-yl] -2-oxo-ethyl acetate using (S) -l, l, l-trifluoro-2-propylamine instead of tert-butylamine. Method B, Rt - 0.92 min., M / z = 424.1 (MH) ', Exact mass: 425.1.^XH NMR (400 MHz, DMSO-d₆) δ ppm 1.19 - 1.40 (m, 3H), 1.92 - 2.30 (m, 2H), 3.08 - 3.27 (m, 1H), 3.37 - 4.03 (m, 4H), 4.47 - 4.78 (m, 1 H), 7.20 (m, J = 54.4 Hz, 1 H), 7.29 - 7.41 (m, 1 H), 7.55 7.80 (m, 1 H), 7.86 - 8.04 (m, 1 H), 9.25 (sa, 1 H), 10.30-10.40 (m, 1H) as a mixture of rotamers.

Compound 31: (3S) -N-r3- (difluoromethyl) -4-fluoro-phenyl1-lT2- (isopropylamino) -2-oxoacetylpyrrolidin-3-carboxamide

<img file="MX2016005342A_D0053.tif" />

<img file="MX2016005342A_D0054.tif" />

<img file="MX2016005342A_D0055.tif" />

Compound 31 was prepared in a similar manner to that described for compound 30 using isopropylamine instead of (S) -l, l, l-trifluoro-2-propylamine. Method B, Rt = 0.83 min., M / z = 370.2 (Μ-H) ', Exact mass; 371.1.^XH NMR (400 MHz, DMSO-d₅) δ ppm 0.75 - 1.42 (m, 6H), 1.95 - 2.29 (m, 2H), 3.05 - 3.26 (m, 1H), 3.36 - 4.04 (m, 5H), 7.20 (m, J = 54.1 , l H), 7.28 - 7.37 (m, 1 H), 7.63 - 7.78 (m, 1 H), 7.87 - 8.03 (m, 1 H), 8.40-8.50 (m, 1 H), 10.25-10.41 (m , 1 H) as a mixture of rotamers.

Compound 32: (3S1-l-r2-oxo-2-rr (lR ') - 2,2,2-tr¡fluoro-l-metíl-etíl1am¡no1acet¡l1-N (3,4, 5-tr¡fluorophenyl) p¡rrol¡d¡n-3-carboxam¡da

Boc- (3S) -l-pyrrolidin-3-carboxylic acid (1.5g, 6.97mmol) and 3,4,5trifluoroaniline (2.51g, 17.05mmol) and HATU (3.18g, 8.36mmol) were dissolved in DMF (5ml ). To this was added Ν, Ν-diisopropylethylamine (3.6 ml, 0.75 g / ml, 20.91 mmol). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was loaded onto a column and purified by silica gel column chromatography using a gradient elution from heptane to EtOAc (from 100: 0 to 0: 100). The desired fractions were concentrated in vacuo to provide tere-butyl (3S) -3 - [(3,4,5-trifluorophenyl] carbamoyl] prolidin-1-carboxylate (2.32 g). Method B, Rt = 1.13 min., M / z - 343.1 (MH), Exact mass: 344.1. HCI (6 M in PrOH, 10 ml, 6 M, 60 mmol) was added to (3S) -3 - [(3,4,5-trifluorophenyl) carbamoyl] p¡rrolidin-l-carboxylate of tere- butyl (2.3 g, 6.35 mmol) in CH₂CI₂ (50 ml) and stirred at room temperature for 5 days. The reaction mixture was concentrated. The residue was brought to CH₂CI₂ (40 ml) and a white precipitate formed which was collected on a glass filter and dried in a vacuum oven at 55 ° C to provide (3S) -N- (3,4,5-trifluorophenyl) p hydrochloride Rrolidin-3-carboxamide (1600 mg) as a bright white powder that was used as such. Method B, Rt = 0.69 min., M / z = 243.0 (MH), Exact mass: 244.1.

Ethyl 2-chloro-2-oxo-acetate (1.98 ml, 1.22 g / ml, 17.69 mmol) was added to a solution of (R) -l, l, l-trifluoro-2-propylamine (2 g, 17.69 mmol ) and triethylamine (4.9 ml, 35.37 mmol) in CH₂CI₂ (20 mi). The reaction mixture was stirred for 1 hour. NaOH (1M in H) was added₂O) (26.5 ml, 1M, 26.53 mmol) and the reaction mixture was stirred vigorously for 2 hours. The organic phase was removed and the aqueous phase was acidified with HCI. Compounds were extracted with diethyl ether (4 x 25 ml). The combined organic phases were dried with Na₂SW₄, filtered and evaporated to dryness resulting in 2-oxo-2 - [[(lR) -2,2,2-tr¡fluoro-l-methyl-ethyl] amino] acetic acid (2.72 g ) as a white powder.

(3S) -N- (3,4,5-Trifluorophenyl) pyrrolidin-3-carboxamide hydrochloride (200 mg) and 2-oxo-2 acid - [[(lR) -2,2,2-tr! fluoro-l-methyl-ethyl] amino] acetic (118 mg, 0.64 mmol) in DMF (2 ml). HATU (266.74 mg, 0.7 mmol) and DIPEA (0.44 ml, 0.75 g / ml, 2.55 mmol) were successively added. The reaction mixture was stirred at room temperature. The reaction mixture was loaded onto a column and purified using silica gel column chromatography (0 to 100% ethyl acetate in heptane a) to provide compound 32 (83 mg) as a white powder. Method B, Rt = 1.04 min., M / z = 410.1 (MH) ', Exact Mass: 411.1. Differential scanning calorimetry: melting point at 197.3 ° C (from 30 to 300 ° C at 10 ° C / min).^XH NMR (400 MHz, DMSO-d₆) δ ppm 1.30 (d, J = 7.0 Hz, 3H), 1.92 - 2.30 (m, 2H), 3.09 - 3.26 (m, 1H), 3.38 - 3.99 (m, 4H), 4.50-4.70 ( m, 1H), 7.40-7.60 (m, 2H), 9.20-9.31 (m, 1H), 10.42-10.49 (m, 1H) as a mixture of rotamers.

Compound 33: (25) -Μ (3- &lt;ΙθΓθ-4,5-όίΑυθΓθί6ηίΙ) -2-ηΓΐ6ίίΙ-1-Γ2-οχο-2-Γ (Ί /?) 2,2,2trifluoro-l-met¡let L) amino1acet¡llpirrolid¡n-3-carboxamida

<img file="MX2016005342A_D0056.tif" />

<img file="MX2016005342A_D0057.tif" />

Methyl (2S, 3S) -l- (2-ethoxy-2-oxo-acetyl) -2-methyl-pyrrole-3-carboxylate (2200 mg, 9.04 mmol) was cooled in methanol (50 my) in an ice water bath. NaOH (1M in H) was added dropwise₂O) (9.95 ml, 1M, 9.95 mmol) and the mixture was stirred for 30 minutes. The reaction was quenched with HCI (1M in H₂O) (9.5 ml, 1M, 9.5 mmol) and concentrated to preserve 20 ml of residue. The residue was extracted with 2-methyl-THF (2 x 20 ml). The combined organic phases were dried (Na₂SW₄) and evaporated to dryness to give 2 - [(25,35) -3-ΓηεΙοχϊθ3Γ0οηιΤ2methyl-pyrrolidin-l-yl] -2-oxo-acetic acid (1930 mg) as a light yellow solid.

A solution of 2 - [(2S, 3S) -3-methoxycarbonyl-2-methyl-pyrrolidin-l-yl] 2-oxo-acetic acid (800 mg, 3.64 mmol) in DMF (4 mL, 54.44 mmol) was cooled and (R) -l, l, l-trifluoro-2-propylamine (494 mg, 4.37 mmol) up to 0 ° C in an ice-water bath. HATU (1524 mg, 4.01 mmol) was then added while continuing to cool. The reaction mixture was stirred at 0 ° C for 30 minutes and allowed to reach room temperature for 1 h. The reaction mixture was loaded on a column and purified using silica gel column chromatography (0 to 100% ethyl acetate in heptane a) to give (2S, 3S) -2-methyl-l- [ 2-oxo-2 - [[(lR) -2,2,2ΡΊΑυοΓΟ-Ι-ΓηεΡΤείΐΠθΓηίηοΙθοείίΙΙρίπΌΐκΙίη-Β-οθΓόοχϊΙβίο of methyl (1000 mg) as a colorless oil. Method D, Rt = 1.59 min., M / z - 309.3 (MH) ', Exact mass: 310.1.

(2S, 3S) -2-Methyl-l- [2-oxo-2 - [[(lR) -2,2,2-trifluoro-l-methylethyl] amino] acetyl] p¡rrol¡d¡n was stirred. -3-methyl carboxylate (400 mg, 1.29 mmol) in methanol (10 ml) at room temperature. NaOH (1 M in H) was added dropwise₂O) (1.35 ml, 1M, 1.35 mmol) and the mixture was stirred for 20 hours. After 20 hours, more NaOH (1M in H) was added₂O) (0.26 mi, 1 M,

0.26 mmol) to the reaction mixture which was stirred at room temperature for 2 hours. The reaction was quenched with HCI (1M in H₂O) (1.61 ml, 1M, 1.61 mmol) and concentrated to preserve 3 ml of residue. The residue was extracted with 2-methyl-THF (2 x 20 ml). The combined organic phases were dried (Na₂SW₄) and evaporated to dryness to give (2S, 3S) -2-methyl-l- [2-oxo-2 [[(lR) -2,2,2-trifluoro-l-methyl-et-l acid ] amino] acetyl] pyrroletin-3-carboxylic acid (440 mg) as a white solid after standing.

A solution of (2S, 3S) -2-methyl-l- [2-oxo-2 - [[(lR) -2,2,2-tr¡fluoro-lmethylethyl] amino] acetyl was cooled. ] prrolidin-3-carboxylic acid (190 mg, 0.64 mmol) in DMF (2 ml) and 3-chloro-4,5-difluoroaniline (115.4 ml, 0.71 mmol) up to 0 ° C in an ice-water bath. Then HATU (292.6 mg, 0.77 mmol) was added while continuing to cool. The reaction mixture was stirred at 0 ° C for 30 minutes and allowed to reach room temperature for 24 h. The reaction mixture was loaded onto a column and purified using silica gel column chromatography (0 to 100% ethyl acetate in heptane a) and further by preparative HPLC (stationary phase: RP XBridge Prep C18 OBD- 10 pm, 30x150 mm; mobile phase: NH dissolution₄HCO₃ 0.25% in water, CH₃CN) resulting in compound 33a (40mg) and compound 33b (33mg). 33a: (2S, 3R) - / V- (3-chloro-4,5-difluorophenyl) -2-methyl-l- [2-oxo-2 - [[(l /?) - 2.2, 2-trifluoro-methylmethyl] amino] acetyl] pyrroletin-3-carboxyamide Method B, Rt - 1.06 min., M / z = 440.1 (MH) ', Exact mass: 441.1. 33b: (2S, 3S) - / V- (3-chloro-4,5-difluorophenyl) -2-methyl-l- [2-oxo-2 - [[(l /?) - 2,2,2- trifluoro-methylmethyl] amino] acetyl] pyrrolidin-3-carboxamide Method B, Rt = 1.11 min., m / z = 440.1 (MH) ', Exact mass: 441.1.^XH NMR (400 MHz, DMSO-d₆) δ ppm 0.99 - 1.05 (m, 3H), 1.26 - 1.34 (m, 3H), 1.95 2.06 (m, 1H), 2.23 - 2.39 (m, 1H), 3.11 - 3.27 (m, 1H) ), 3.38 - 3.84 (m, 2H), 4.46 - 4.87 (m, 2H), 7.60 - 7.69 (m, 2H), 9.17 - 9.43 (m, 1H), 10.24 - 10.51 (m, 1H) ) as a mixture of rotamers

Compound ________ 34: ________ (2S) -2-methyl-lT2-oxo-2Tr (lR) -2,2,2-trifluoro-l-methylethylamino1acet¡l1-N- (3,4,5-tr¡fluorophenyl) pyrrolid N-3-carboxamide

<img file="MX2016005342A_D0058.tif" />

Compound 34a (44 mg) and 34b (52 mg) were prepared in a similar manner to that described for compound 33a and 33b, using 3,4,5-trifluoroaniline instead of 3-chloro-4,5-methylaniline. 34a: (2S, 3R) -2-methyl-l- [2-oxo-2 - [[(lR) -2,2,2-trifluoro-l-methyl-ethyl] amino] acetyl ] -N (3,4,5-trifluorophenyl) pyrrolidin-3-carboxamide Method B, Rt = 1.02 min., M / z = 424.1 (MH) ', Exact mass: 425.1. 34b: (2S, 3S) -2-methyl-l- [2-oxo-2 - [[(lR) -2,2,2-tr¡fluoro-l-methyl-ethyl] amino] acet L] -N (3,4,5-trifluorophenyl) pyrrolidin-3-carboxamide Method B, Rt = 1.05 min., M / z - 424.1 (MH) ', Exact mass: 425.1. NMR (400 MHz, DMSO-d₆) δ ppm 0.99 - 1.05 (m, 3H), 1.26 - 1.34 (m, 3H), 1.92 2.07 (m, 1H), 2.19 - 2.41 (m, 1H), 3.08 - 3.28 (m, 1H) ), 3.38 - 3.85 (m, 2H), 4.45 - 4.87 (m, 2H), 7.43 - 7.57 (m, 2H), 9.30 (bs, 1H), 10.41 (bs, 1H) as a mixture of rotamers.

<td>Compound 35:</td><td>/ V- (3-chloro-2,4-difluorophenyl) -2-methyl-ir2-oxo-2-r ((l /?) - 2,2,2-</td>

tr¡fluoro-l-met¡let¡l) am¡nolacetillp¡per¡d¡n-3-carboxam¡da

<img file="MX2016005342A_D0059.tif" />

Ethyl l - ^ - ethoxy ^ -oxo-acetyl ^ -methyl-piperidin-S-carboxylate was prepared from ethyl 2-methylpiperin-3-carboxylate, similarly as described for (2S, Methyl 3S) -l- (2-ethoxy-2-oxoacetyl) -2-methyl-pyrrolidin-3-carboxylate from methyl (2S, 3S) -2-methylpyrrolidin-3-carboxylate. Compound 35 was prepared in a similar manner to that described for compound 33, starting from ethyl l- (2-ethoxy-2-oxo-acetyl) -2-methyl-piperidin-3-carboxylate instead of (2S, 3S) -l- (2-ethoxy-2-oxo-acetyl) -2-methyl-methylprolidin-3-carboxylate and using 3-doro-2,4-difluoro-aniline instead of 3-chloro-4,5-difluoroaniline. Compound 35 (550 mg) was separated into diastereoisomers 35a, 35b, 35c and 35d by preparative SFC (stationary phase: Chiralpak Daicel IC 20 x 250 mm; mobile phase: CO₂, EtOH with 0.2% PrNH₂). Compound 35a ((2S, 3S) or (2R, 3R), first eluent in SFC, 70 mg), Method D, Rt = 1.86 min., M / z = 454.1 (MH) ', Exact mass: 455.1. Compound 35b ((2S, 3S) or (2R, 3R), second eluent in SFC, 88 mg) Method D, Rt - 1.87 min., m / z = 454.1 (MH) ', Exact Mass: 455.1.

Compound 35c ((2S, 3R) or (2R, 3S), third eluent in SFC, 86 mg), Method D, Rt = 1.89 min., M / z = 454.1 (MH), Exact mass: 455.1. Compound 35d ((2S, 3R) or (2R, 3S), fourth eluent in SFC, 106 mg), Method D, Rt - 1.88 min., M / z - 454.1 (MH), Exact mass: 455.1.

BIOLOGICAL EXAMPLES anti-HBV activity of the compounds of formula II)

Anti-HBV activity was measured using a stable transfected cell line,

HepG2.2.15. This cell line has been reported to secrete relatively uniformly high levels of HBV virion particles, which have been shown to cause both acute and chronic infection and disease in chimpanzees.

For the antiviral assay, cells were treated twice for three days with the serially diluted compound in 96-well plates in duplicate. After 6 days of treatment, antiviral activity was determined by quantifying purified HBV DNA from secreted virions using real-time PCR and a HBV-specific probe and primer set.

Anti-HBV activity was also measured using the HepG2.117 cell line, a stable cell line that produces HBV in an induced manner, which replicates HBV in the absence of doxycycline (Tet-off system). For the antiviral assay, HBV replication was induced, followed by treatment with the serially diluted compound in duplicate 96-well plates. After 3 days of treatment, antiviral activity was determined by quantifying intracellular HBV DNA using real-time PCR and a HBV-specific probe and primer set.

The cytotoxicity of the compounds was tested using HepG2 cells, incubated for 4 days in the presence of the compounds. Cell viability was evaluated using a resazurin assay. The results are presented in Table 1.

TABLE 1

<td>Comp. no.</td><td>HepG2 2.15 EC₅₀im)</td><td>HepG2 117 CE₅₀(p.m)</td><td>HepG2 4 days CC₅₀ (p.m)</td>

<td> 1</td><td> 0.020</td><td> 0.018</td><td> &gt;25</td>

<td> 2</td><td> 0.070</td><td> 0.033</td><td> &gt;25</td>

<td> 3</td><td> 0.141</td><td> 0.026</td><td> &gt;25</td>

<td> 4</td><td> 0.126</td><td> 0.071</td><td> &gt;25</td>

<td> 5</td><td> 0.112</td><td> 0.046</td><td> &gt;25</td>

<td> 6</td><td> 0.301</td><td> 0.257</td><td> &gt;25</td>

<td> 7</td><td> 0.067</td><td> 0.117</td><td> &gt;25</td>

<td> 8</td><td> 0.065</td><td> 0.038</td><td> &gt;25</td>

<td> 9</td><td> 0.120</td><td> 0.134</td><td> &gt;25</td>

<td> 10</td><td> 0.008</td><td> 0.009</td><td> &gt;25</td>

<td> 11</td><td> 0.032</td><td> 0.017</td><td> &gt;25</td>

<td> 12</td><td> 0.321</td><td> 0.115</td><td> &gt;25</td>

<td> 13</td><td> 0.020</td><td> 0.035</td><td> &gt;25</td>

<td> 14</td><td> 0.064</td><td> 0.045</td><td> &gt;25</td>

<td> 15</td><td> 0.025</td><td> 0.047</td><td> &gt;25</td>

<td> 16</td><td> 0.058</td><td> 0.035</td><td> &gt;25</td>

<td>17a</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>

TABLE 1

<td>17b</td><td> 0.918</td><td> 0.796</td><td> &gt;25</td>

<td>17c</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>

<td>17d</td><td> 0.070</td><td> 0.032</td><td> &gt;25</td>

<td> 18</td><td></td><td> 0.670</td><td> &gt;25</td>

<td> 19</td><td> 0.496</td><td> 0.449</td><td> &gt;25</td>

<td> 20</td><td> 0.289</td><td> 0.645</td><td> &gt;25</td>

<td> 21</td><td> 0.063</td><td> 0.063</td><td> &gt;25</td>

<td> 22</td><td> 0.110</td><td> 0.128</td><td> &gt;25</td>

<td> 23</td><td> 0.380</td><td> 0.575</td><td> &gt;25</td>

<td> 24</td><td> 0.134</td><td> 0.384</td><td> &gt;25</td>

<td> 25</td><td> 0.042</td><td> 0.031</td><td> &gt;25</td>

<td> 26</td><td> 0.168</td><td> 0.122</td><td> &gt;25</td>

<td> 27</td><td> 0.119</td><td> 0.126</td><td> &gt;25</td>

<td> 28</td><td> 0.050</td><td> 0.083</td><td> &gt;25</td>

<td>29a</td><td> 0.010</td><td> 0.011</td><td> &gt;25</td>

<td>29b</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>

<td> 29</td><td> 0.018</td><td> 0.048</td><td> &gt;25</td>

<td> 30</td><td> 0.161</td><td> 0.125</td><td> &gt;25</td>

<td> 31</td><td> 0.134</td><td> 0.143</td><td> &gt;25</td>

<td> 32</td><td></td><td> 0.052</td><td> &gt;25</td>

<td>33a</td><td></td><td> &gt;0.5</td><td> &gt;25</td>

<td>33b</td><td></td><td> 0.005</td><td> &gt;25</td>

<td>34a</td><td></td><td> &gt;0.5</td><td> &gt;25</td>

<td>34b</td><td></td><td> 0.004</td><td> &gt;25</td>

<td>35a</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>

<td>35b</td><td> 0.195</td><td> 0.483</td><td> &gt;25</td>

<td>35c</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>

<td>35d</td><td> &gt;1</td><td> &gt;1</td><td> &gt;25</td>