RU2300521C2 - Novel derivatives of mandelic acid and their using as thrombin inhibitors - Google Patents

Abstract

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel compounds chosen from the following substances: Ph-(3-Cl)-(5-OCHF₂)-(R)-CH(OH)C(O)-Aze-Pab(OMe); Ph-(3-Cl)-(5-OCHF₂)-(R)-CH(OH)C(O)-Aze-Pab(OH), and Ph-(3-Cl)-(5-OCHF₂)-(R)-CH(OH)C(O)-Aze-Pab or their pharmaceutically acceptable salts. Proposed compounds inhibit thrombin and can be used in pharmaceutical composition for treatment state wherein thrombin inhibition is required.

EFFECT: valuable medicinal properties of compounds.

16 cl, 49 ex

Description

Field of Invention

This invention relates to new pharmaceutically useful compounds, in particular to compounds that are competitive inhibitors of trypsin-like serine proteases, in particular thrombin, or compounds that are metabolized to such competitive inhibitors, to their use as medicines containing their pharmaceutical compositions and preparation them by synthesis.

BACKGROUND OF THE INVENTION

Blood coagulation is a key process involved in both hemostasis (i.e. preventing the loss of blood from a damaged vessel) and thrombosis (i.e. the formation of a blood clot in a blood vessel, sometimes leading to blockage of the vessel).

Coagulation is the result of a complex group of enzymatic reactions. One of the final stages in this group of reactions is the conversion of the prothrombin proenzyme to the active thrombin enzyme.

Thrombin is known to play a central role in coagulation. It activates platelets, leading to platelet aggregation, converts fibrinogen into fibrin monomers, which spontaneously polymerize into fibrin polymers, and activates factor XIII, which in turn crosslinks these polymers to form insoluble fibrin. In addition, thrombin activates factor V and factor VIII, leading to the formation of thrombin from prothrombin by the "positive feedback" mechanism.

It is expected that, by inhibiting platelet aggregation and the formation and crosslinking of fibrin, effective thrombin inhibitors will exhibit antithrombotic activity. In addition, antithrombotic activity is expected to be enhanced by effective inhibition of the positive feedback mechanism.

State of the art

An earlier development of low molecular weight thrombin inhibitors was described by Claesson in Blood Coagul. Fibrinol. (1994) 5, 411.

Blombäck et al. (in J. Clin. Lab. Invest. 24, suppl. 107, 59, (1969)) thrombin inhibitors based on the amino acid sequence located around the fibrinogen Aα cleavage site have been reported. Regarding the discussed amino acid sequences, these authors suggested that the tripeptide sequence Phe-Val-Arg (P9-P2-P1, hereinafter referred to as the P3-P2-P1 sequence) would be the most effective inhibitor.

Thipebin inhibitors based on dipeptidyl derivatives with α, ω-aminoalkylguanidine in position P1 are known from US patent No. 4346078 and international patent application WO 93/11152. Similar, structurally related dipeptidyl derivatives have also been reported. For example, international patent application WO 94/29336 discloses compounds, for example, with aminomethylbenzamidines, cyclic aminoalkylamidines and cyclic aminoalkylguanidines at position P1 (international patent application WO 97/23499 discloses prodrugs of some of these compounds); European patent application 0648780 discloses compounds, for example, with cyclic aminoalkylguanidines at position P1.

Peptidyl derivative thrombin inhibitors also having as part cyclic aminoalkylguanidines (e.g., either 3- or 4-aminomethyl-1-amidinopiperidine) at position P1 are known from European patent applications 0468231, 0559046 and 0641779.

Thrombin inhibitors based on tripeptidyl derivatives with arginine aldehyde at position P1 were first disclosed in European patent application 0185390.

Later, peptidyl derivatives based on arginine aldehyde modified at the position of the RE were reported. For example, international patent application WO 93/18060 discloses hydroxy acids, European patent application 0526877 - desamino acids and European patent application 0542525 - O-methylmindal acids in position P3.

Inhibitors of serine proteases (e.g., thrombin) based on electrophilic ketones at position P1 are also known. For example, European Patent Application 0195212 discloses peptidyl-α-ketoesters and amides, European Patent Application 0362002 discloses fluoroalkylamide ketones, European Patent Application 0364344 discloses α, β, δ-triketo compounds and European Patent Application 0530167 discloses α-alkoxy ketones in position P1.

Other structurally different trypsin-like serine protease inhibitors based on the C-terminus of arginine derivatives and their isothiouronium analogues are known from European Patent Application 0293881.

More recently, peptidyl derivative thrombin inhibitors have been described in European Patent Application 0669317 and International Patent Applications WO 95/35309, WO 95/23609, WO 96/25426, WO 97/02284, WO 97/46577, WO 96/32110, WO 96/31504, WO 96/03374, WO 98/06740, WO 97/49404, WO 98/57932, WO 99/29664, WO 00/35869 and WO 00/42059.

In particular, WO 97/02284 and WO 00/42059 describe thrombin inhibitors with substituted mandelic acids at position P3.

However, there remains a need for effective trypsin-like serine protease inhibitors such as thrombin. There is also a need for compounds that have a convenient pharmacokinetic profile and are selective in inhibiting thrombin compared to other serine proteases, in particular those involved in hemostasis. Compounds that exhibit the activity of a competitive inhibitor against thrombin are expected to be especially useful as anticoagulants and, therefore, in the therapeutic treatment of thrombosis and related disorders.

Description of the invention

The invention provides a compound of formula I

where R ^a represents —OH or —CH ₂ OH;

R ¹ represents at least one possible halogen substituent;

R ² represents one or two C _1-3 alkoxy substituents, the alkyl parts of these substituents being substituted by one or more fluoro substituents (i.e., R ² represents one or two fluoroalkoxy (C _1-3 ) groups);

Y represents —CH ₂ - or - (CH ₂ ) ₂ - and

R ³ represents a structural fragment of the formula I (1) or I (2)

where R ⁴ represents H or one or more than one fluoro substituent and

one or two of X ₁ , X ₂ , X ₃ and X ₄ are —N— and the rest are —CH—,

or a pharmaceutically acceptable derivative thereof.

The term “pharmaceutically acceptable derivatives” includes pharmaceutically acceptable salts (for example, salts obtained by addition of an acid).

Abbreviations are provided at the end of this specification. The wavy lines on the bonds in the fragments of formulas I (1) and I (2) indicate the attachment positions of these fragments.

Halo groups with which R ¹ may be represented include fluoro, chloro, bromo and iodo. To avoid ambiguity, by saying that R ¹ represents at least one possible halogen group, it is understood that R ¹ may either not be present (and thus be replaced by H, while the valency rules are observed), or it may represent one or more than one halogen atom.

When R ³ is a structural fragment of formula I (1) in which R ⁴ is one or more fluoro substituents, preferred compounds of formula I include those in which R ⁴ is a single fluoro substituent at position 2 or 3 or two fluoro substituents or at positions 2 and 5, or, more preferably, at positions 2 and 6 (where the positions of the substituents are determined relative to the point of attachment of the structural fragment of formula I (1) to the rest of the molecule (i.e. to the —NHCH ₂ - group)).

When R ³ is a structural fragment of formula I (2), preferred compounds of formula I include those in which either

(a) one of X ₁ , X ₂ , X ₃ and X ₄ represents —N—, and the rest represents —CH—, or

(b) both X ₁ and X ₃ or both X ₂ and X ₄ are —N— and the other two (as appropriate) are —CH—.

Preferred compounds of formula I include those in which

R ¹ represents a single fluoro, chloro or bromo substituent;

R ² is C _1-2 alkoxy substituted with one or more fluoro substituents, such as —OCHF ₂ , —OCF ₃ , —OCH ₂ CF ₃ , —OCH ₂ CHF ₂ , —OCH ₂ CH ₂ F, or - OCH (CH ₂ F) ₂ ;

R ³ represents a structural fragment of formula I (1);

R ⁴ represents N.

More preferred compounds of formula I include those in which

R ^a represents OH;

R ¹ represents a single chloro substituent;

R ² represents —OCF ₂ , preferably —OCH ₂ CHF ₂ or, more preferably, —OCHF ₂ or —OCH ₂ CH ₂ F.

Preferred substitution points of R ¹ and R ² on the corresponding phenyl group of the compound of formula I include both meta positions relative to the point of attachment of this phenyl group to the rest of the molecule (i.e., at position 3 and / or 5 (preferably 3,5 substitution) relative to carbon an atom carrying an α- or β-hydroxy acid group).

Compounds of formula I that may be mentioned include those in which

R ² represents one or two C _1-2 alkoxy substituents, the alkyl parts of which are themselves substituted by one or more fluorine substituents, or

R ² represents one or two C ₃ alkoxy substituents, the alkyl parts of which are themselves substituted by one or more fluorine substituents.

The compounds of formula I can be obtained in accordance with methods well known to specialists in this field of technology, for example, as described below.

According to another aspect of the present invention, a method for producing compounds of formula I, wherein

(1) carry out the combination of compounds of formula II

where R ^a , R ¹ and R ² are as defined above, with a compound of formula III

where Y and R ³ are as defined above, for example, in the presence of a coupling agent (e.g., oxalyl chloride in DMF, EDC, DCC, HBTU, HATU, RuBOP or TBTU), an appropriate base (e.g., pyridine, DMAP, TEA, 2, 4,6-collidine or DIPEA) and a suitable organic solvent (e.g. dichloromethane, acetonitrile, EtOAc or DMF);

(2) carry out the combination of compounds of formula IV

where R ^a , R ¹ , R ² and Y are as defined above, with a compound of formula V

where R ³ is as defined above, for example, under the conditions described above in method (1);

(3) for compounds of formula I in which R ¹ is not present, the corresponding compound of formula Ia is reduced as defined above, in which R ⁵ is OR ⁶ , where R ⁵ and R ⁶ are as defined above, for example, by hydrogenation in the presence of a suitable catalyst (e.g., a supported metal catalyst, such as Pd / C (e.g. 10% (w / w) Pd / C)) and a suitable solvent (e.g. lower (e.g. C _1-6 ) alkyl alcohol, such as ethanol), and possibly in the presence of a suitable acid (e.g., acetic acid oty), and / or as described in Synth. Comm. (1998) 4351, or

(4) reacting the corresponding compound of formula XVIA or XVIB, as defined below, with a suitable source of ammonia (e.g., ammonium acetate or gaseous ammonia) under conditions known to those skilled in the art, for example, by reacting an ethylimidoate intermediate (formed in by reacting a compound of formula XVIA or XVIB with HCl (g) in ethanol) with gaseous ammonia in ethanol or under the conditions described in Tetrahedron Lett. 40, 7067 (1999), the disclosure of this document is incorporated herein by reference (for example, to obtain compounds of formula I in which R ³ is a structural fragment of formula I (2) in which X ² or X ⁴ is N, the interaction of the corresponding compounds of formula XVIB with ammonium acetate (for example, with 1-30 equivalents of ammonium acetate) in the presence of N-acetylcysteine (for example, 1-30 equivalents of N-acetylcysteine) and the corresponding solvent (for example, lower alkyl (C _1-6 ) alcohol such as methanol)).

Compounds of formula II are available by using known and / or standard techniques.

For example, compounds of formula II in which R ³ is OH can be prepared by reacting an aldehyde of formula VI

where R ¹ and R ² are as defined above, with

(a) a compound of formula VII

where R "represents H or (CH ₃ ) ₃ Si, for example, at room or elevated temperature (for example, below 100 ° C) in the presence of a suitable organic solvent (for example, chloroform or methylene chloride) and, if necessary, in the presence of a suitable base (e.g., TEA) and / or a suitable catalyst system (e.g., benzylammonium chloride or zinc iodide or using a chiral catalyst, e.g. as described in Chem. Rev., (1999) 99, 3649) followed by hydrolysis under conditions that are good known to specialists in this area parts of technology (for example, as described below);

(b) NaCN or KCN, for example, in the presence of NaHSO ₃ and water, followed by hydrolysis;

(c) chloroform, for example, at elevated temperature (for example, above room temperature, but below 100 ° C) in the presence of a suitable organic solvent (e.g. chloroform) and, if necessary, in the presence of a suitable catalyst system (e.g. benzylammonium chloride) followed by hydrolysis ;

(g) a compound of formula VIII

where M represents Mg or Li, followed by oxidative cleavage (for example, ozonolysis or cleavage catalyzed by osmium or ruthenium) under conditions that are well known to specialists in this field of technology, or

(e) Tris (methylthio) methane under conditions well known to those skilled in the art, followed by hydrolysis in the presence of, for example, HgO and HBF ₄ .

Compounds of formula II in which R ^a is —CH ₂ OH can be prepared by reducing a compound of formula IX

where R is C _1-6 alkyl or C _1-3 alkyl phenyl and R ¹ and R ² are as defined above, for example, at room or lower temperature in the presence of a suitable reducing agent (e.g. sodium borohydride) and an appropriate organic solvent ( e.g. methanol, ethanol, THF or mixtures thereof), followed by hydrolysis of the resulting tropic acid ester intermediate of formula IXA

where R, R ¹ and R ² are as defined above, under conditions well known to those skilled in the art, for example, as described below. One skilled in the art will recognize that the reduction and hydrolysis steps can be performed in one tank, for example, as described below.

Compounds of formula II in which R ^a is —OH may alternatively be prepared by oxidizing a compound of formula IXB

or a derivative thereof, which is optionally protected by a secondary hydroxyl group, where R ¹ and R ² are as defined above, in the presence of a suitable oxidizing agent (e.g., a combination of a suitable free radical oxidizing agent (such as TEMPO) and an appropriate hypochlorite salt (such as sodium hypochlorite) ) under conditions known to those skilled in the art, for example at a temperature between -10 ° C and room temperature in the presence of a suitable solvent (e.g. water, acetone or a mixture thereof), an appropriate salt (e.g., g alkali metal halides such as potassium bromide) and a suitable base (e.g., alkali metal carbonate or hydrogencarbonate such as sodium hydrogencarbonate).

Enantiomeric forms of a compound of formula II in which R ^a is —OH (that is, those compounds that have different configurations of substituents around the C atom at the α position relative to the CO ₂ H group) can be separated using the step of forming the enantiospecific derivatives. This can be done, for example, in an enzymatic manner. Such enzymatic methods include, for example, transesterification of the α-OH group at a temperature between room temperature and reflux temperature (e.g. between 45 and 65 ° C) in the presence of a suitable enzyme (e.g. PS Amano Lipase), corresponding ester (e.g. vinyl acetate) and a suitable solvent (e.g. methyl tert-butyl ether). An isomer whose derivative is formed can then be separated from the unreacted isomer using conventional separation techniques (e.g., chromatography).

Groups added to compounds of formula II at this derivative formation stage can be removed either prior to any further reactions or at any later stage in the synthesis of compounds of formula I. Additional groups can be removed using conventional techniques (e.g., for esters α-OH groups — by hydrolysis under conditions known to those skilled in the art (e.g., at a temperature between room temperature and reflux temperature in the presence of a suitable base (e.g., NaOH) and correspondingly solvent (e.g. MeOH, water, or mixtures thereof))).

The enantiomeric forms of the compound of formula II in which R ^a is —CH ₂ OH can be separated using chiral chromatography techniques (eg, chiral HPLC).

Compounds of formula III can be prepared by coupling a compound of formula X

where Y is the same as defined above with a compound of formula V as defined above, for example, under conditions similar to those described herein for the preparation of compounds of formula I.

Compounds of formula IV can be prepared by combining a compound of formula II as defined above with a compound of formula X as defined above, for example, under conditions similar to those described herein for the preparation of compounds of formula I.

Compounds of formula VI are available using known and / or standard techniques. For example, they can be obtained in the following way:

(1) metallization (where the metal may be, for example, an alkali metal such as Li, or preferably a divalent metal such as Mg) of a compound of formula XI

where Hal represents a halogen atom selected from Cl, Br and I, and R ¹ and R ² are as defined above, followed by reaction with a suitable source of the formyl group (for example, N, N-dimethylformamide), for example, under conditions described below;

(2) reduction of the compounds of formula XII

where R ¹ and R ² are as defined above, in the presence of a suitable reducing agent (for example, DIBAL-H), or

(3) oxidizing a compound of formula XIII

where R ¹ and R ² are as defined above, in the presence of a suitable oxidizing agent (for example, MnO ₂ , pyridinium chlorochromate, a combination of DMSO and oxalyl chloride, or the SO ₃ pyridine complex in DMSO).

The compounds of formula IX can be obtained from the corresponding phenylacetate (which, for example, can be obtained from the corresponding acetophenone, as described in J. Am. Chem. Soc. 98, 6750 (1976), or from the corresponding benzyl cyanide using standard hydrolytic procedures) with using traditional techniques, for example, by analogy with the methods described in J. Org. Chem. 54, 3831 (1989), and / or as described below.

Compounds of formula IX can be prepared by dihydroxylation of the corresponding compound of formula XIIIA

where R ¹ and R ² are as defined above in the presence of a suitable dihydroxylating agent (for example, a reagent or a mixture of reagents that give OsO ₄ , such as AD-mix-α or, in particular, AD-mix-β), for example under conditions known to those skilled in the art, for example at a temperature between -10 ° C and room temperature in the presence of an appropriate solvent (for example, water, tert-butanol or a mixture thereof). If asymmetric oxidizing agents such as AD-mix-α or AD-mix-β are used, this method can be used to obtain compounds of formula IXB having certain configurations (i.e., R or S) of groups around C atoms to which primary and secondary hydroxyl are attached groups.

Compounds of formula XIIIA can be prepared by reacting the corresponding compound of formula XI, as defined above, with a suitable vinyl anion source (e.g., tributyl (vinyl) tin) under conditions known to those skilled in the art, for example, between room temperature and reflux temperature (e.g. 50 ° C) in the presence of an appropriate solvent (e.g. toluene), a suitable coupling agent (e.g., a palladium (0) coordination complex such as tetrakis (triphenylphosphine) pa lalladium (0)) and possibly in the presence of an appropriate catalyst (e.g., 2,6-di-tert-butyl-4-methylphenol).

Compounds of formulas V, VII, VIII, X, XI, XII and XIII are either commercially available, well known from the literature, or can be obtained by analogy to the methods described herein or using traditional synthesis procedures according to standard procedures from readily available starting materials using appropriate reagents and reaction conditions. Compounds of formulas Ia, XVIA and XVIB can be prepared by the methods described below.

Substituents on the phenyl ring in the compounds of formulas I, II, III, IV, V, VI, IX, IXA, IXB, XI, XII, XIII and XIIIA can be introduced and / or interconverted using methods well known to specialists in this field of technology , by standard interconversion of functional groups according to standard procedures from readily available starting materials using appropriate reagents and reaction conditions.

For example, compounds of formulas I, II, IV, VI, IXA, XI, XII and XIII can be obtained from the corresponding compounds of formulas XIVA, XIVB, XIVC, XIVD, XIVE, XIVF, XIVG and XIVH, respectively

where R ^a , R, R ¹ , R ³ , Y and Hal (as appropriate) are as defined above, for example:

(a) by reacting the corresponding fluorinated haloalkane (e.g., fluorinated chloralane), for example, at room or higher temperature (e.g., reflux temperature) in the presence of a suitable base (e.g. tert-butylate, KOH or NaOH, e.g. aqueous solution) and an appropriate organic solvent (e.g. THF, chloroform or isopropanol) or

(b) as a result of the interaction of the compounds of formula XIVJ

where R ^x represents C _1-4 alkyl, C _1-4 perfluoroalkyl or phenyl (optionally substituted with methyl, nitro or halogen) and R ^y represents CH ₂ CF ₃ , CH ₂ CHF ₂ , CH ₂ CH ₂ F or CH ( CH ₂ F) ₂ , for example, in the presence of a suitable base (e.g. K ₂ CO ₃ ) and an appropriate solvent (e.g. DMF),

for example, in both cases, as described below.

One skilled in the art will understand that such conversions with functional groups can also be carried out at an earlier stage in the complete synthesis scheme for compounds of formulas II, IV, VI, IXA, XI, XII and XIII (i.e., on the corresponding precursors of compounds of formulas XIVB, XIVC, XIVD, XIVE, XIVF, XIVG and XIVH respectively).

Compounds of formulas XIVA, XIVB, XIVC, XIVD, XIVE, XIVF, XIVG, XIVH and XIVJ are either commercially available, known from the literature, or can be obtained either by analogy to the methods described herein or by using traditional synthesis procedures according to standard procedures from available starting materials using appropriate reagents and reaction conditions. For example, compounds of formulas XIVA, XIVB, XIVC, XIVD, XIVE, XIVF, XIVG and XIVH can be obtained by deprotecting the corresponding protected phenols (where the protecting group may be, for example, methyl, allyl, benzyl or tert-butyl) in standard conditions. In addition, compounds of formula XIVD in which R ¹ is a single chloro substituent can be prepared from di- or trihalogenated benzene (e.g., 1-Br-, 3-Cl-, 5-F-benzene by substituting a fluorine atom with a methoxy group (e.g. by reacting with NaOMe in a 1-methyl-2-pyrrolidinone / methanol mixture at elevated temperature), replacing the bromo group with a formyl group (for example, as described above to obtain compounds of formula VI) and then demethylating (for example, using PhSH in 1-methyl -2-pyrrolidinone in the presence of K ₂ CO ₃ )).

In addition, compounds of formula I in which R ^{1 is} absent can be prepared from the corresponding compounds of formula I (or using their respective precursors) in which R ¹ is halogen (such as chloro), for example, by hydrogenation under conditions known to specialists in this field of technology.

Compounds of formula I can be isolated from their reaction mixtures using conventional techniques.

According to the present invention, pharmaceutically acceptable derivatives of compounds of formula I also include “protected” derivatives of compounds of formula I and / or compounds that act as prodrugs.

Compounds that may act as prodrugs of compounds of formula I that may be mentioned include compounds of formula Ia

where R ^3a represents a structural fragment of the formula I (3) or I (4)

where R ⁵ represents OR ⁶ or C (O) OR ⁷ ;

R ⁶ represents H, C _1-10 alkyl, C _1-3 alkylaryl or C _1-3 alkyloxyaryl (moreover, the alkyl parts of the last two groups are possibly interrupted by one or more oxygen atoms and the aryl parts of the last two groups are possibly substituted by one or more than one substituent selected from halogen, phenyl, methyl or methoxy, the last three groups being also optionally substituted with one or more halogen substituents);

R ⁷ is C _1-10 alkyl (wherein the last group is optionally interrupted by one or more oxygen atoms) or C _1-3 alkylaryl or C _{1-3 is} alkyloxyaryl (wherein the alkyl moieties of the last two groups are optionally interrupted by one or more than one atom oxygen and aryl moieties of the last two groups are optionally substituted with one or more substituents selected from halogen, phenyl, methyl or methoxy, the last three groups being also optionally substituted with one or more halogen substituents), and

R ^a , R ¹ , R ² , Y, R ⁴ , X ₁ , X ₂ , X ₃ and X ₄ are as defined above,

and their pharmaceutically acceptable derivatives.

The term “pharmaceutically acceptable derivatives” of compounds of formula Ia includes pharmaceutically acceptable salts (for example, salts obtained by addition of an acid).

Wavy lines on the bonds in the fragments of formulas I (3) and I (4) indicate the attachment positions of these fragments.

Alkyloxyaryl groups, which may be R ⁶ and R ⁷ , include an alkyl and aryl group bonded through an oxygen atom. Alkylaryl and alkyloxyaryl groups are attached to the remainder of the molecule through the alkyl part of these groups, and these alkyl parts (if there are a sufficient number of carbon atoms, i.e. three) can have a branched chain. The aryl portions of the alkylaryl and alkyloxyaryl groups which R ⁶ and R ⁷ may or may be substituted include carbocyclic and heterocyclic aromatic groups such as phenyl, naphthyl, pyridinyl, oxazolyl, isoxazolyl, thiadiazolyl, indolyl, benzofuranyl and the like.

Alkyl groups, which may be R ⁶ and R ⁷ , may be straight chain or, if there is a sufficient number (i.e. at least three) carbon atoms, they may be branched chain and / or cyclic. In addition, if there is a sufficient number (i.e., at least four) of carbon atoms, then such alkyl groups can also be partially cyclic / acyclic. Such alkyl groups may also be saturated or, if there is a sufficient amount (i.e., at least two) of carbon atoms, be unsaturated.

Halo groups with which R ⁶ and R ⁷ may be substituted include fluoro, chloro, bromo and iodo.

When R ⁵ is C (O) OR ⁷ , preferred R ⁷ groups include

(a) linear, branched or cyclic C _3-6 alkyl, for example C _4-6 cycloalkyl;

(b) C _1-2 alkylaryl groups, such as benzyl, optionally substituted as described above.

Preferred compounds of formula Ia include those in which R ⁵ is OR ⁶ .

When R ⁵ is OR ⁶ , preferred R ⁶ groups include

(a) H;

(b) unsubstituted linear, branched or cyclic C _3-8 alkyl (e.g. C _1-6 ), such as linear C _1-3 alkyl (e.g. ethyl or, especially methyl), branched C _3-8 alkyl ( for example iso-propyl, iso-butyl or 4-heptyl) or cyclic C _4-7 alkyl (i.e. C _4-7 cycloalkyl, e.g. cyclobutyl or cyclohexyl);

(c) C _1-3 alkyloxyphenyl (e.g., C ₂ alkyloxyphenyl), wherein the phenyl group is optionally substituted with one or more substituents, as indicated above (e.g., trifluoromethyl);

(d) C _1-2 alkylaryl (e.g. methyl aryl), wherein the aryl group is phenyl, pyridinyl, oxazolyl or isoxazolyl, wherein the last three groups are optionally substituted with one or more substituents as indicated above (e.g. methoxy, methyl, bromo and / or chloro).

Preferred compounds of formula Ia include those in which R ⁵ is OR ⁶ and R ⁶ is linear, branched (as appropriate) or cyclic (as appropriate) C _1-6 alkyl (e.g. C _1-4 ), such as methyl, ethyl, n-propyl, iso-propyl or cyclobutyl.

Compounds of formula Ia can be prepared using one or more of the following methods:

(a) reacting the corresponding compound of formula II, as defined above, with a compound of formula XV

where Y and R ^3a are as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I;

(b) reacting the corresponding compound of formula IV, as defined above, with a compound of formula XVI

where R ^3a is as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I;

(c) for compounds of formula Ia in which R ⁵ is OH, the corresponding compound of formula XVIA or XVIB is reacted

where R ^a , R ¹ , R ² , R ⁴ , Y, X ₁ , X ₂ , X ₃ and X ₄ are as defined above with hydroxylamine, for example, under conditions known to those skilled in the art;

(d) for compounds of formula Ia in which R ⁵ is OR ⁶ , a protected derivative of the corresponding compound of formula I is reacted, which is, for example, a compound of formula XVII

where R ^3b represents a structural fragment of the formula I (5) or I (6)

where R ^b represents, for example, -CH ₂ CH ₂ -Si (CH ₃ ) ₃ or benzyl, or a tautomer thereof, and R ^a , R ¹ , R ² , Y, R ⁴ , X ₁ , X ₂ , X ₃ and X ₄ are as defined above with a compound of formula XVIII

where R ⁶ is as defined above or its salt obtained by addition of an acid, for example, at a temperature between room temperature and reflux temperature in the presence of an appropriate organic solvent (e.g. THF, CH ₃ CN, DMF or DMSO), followed by removal of the group -C ( O) OR ^b under conditions known to those skilled in the art (for example, by reacting with QF or TFA (for example, as described below));

(e) for compounds of formula Ia in which R ⁵ is OH, a compound of formula XVII, as defined above, is reacted in which R ^b is benzyl with hydroxylamine or a salt thereof obtained by addition of an acid, for example, under conditions which known to specialists in this field of technology;

(e) for compounds of formula Ia in which R ⁵ is COOR ⁷ , the corresponding compound of formula I, as defined above, is reacted with a compound of formula XIX

where L ¹ represents a suitable leaving group, such as halogen or nitrophenyl (for example, 4-nitrophenyl), and R ⁷ as defined above, for example, at approximately room temperature in the presence of a suitable base (for example, NaOH, for example, in aqueous solution) and an appropriate organic solvent (e.g. methylene chloride) or

(g) for compounds of formula Ia in which R ⁵ is OCH ₃ or OCH ₂ CH ₃ , the corresponding compound of formula Ia in which R ⁵ is OH is reacted with dimethyl sulfate or diethyl sulfate, respectively, for example, in the presence of a suitable base ( for example, an alkali metal hydroxide such as KOH (for example, in an aqueous solution at a concentration of, for example, 50 wt.%) and an appropriate catalyst (for example, a quaternary ammonium halide, such as benzyltrimethylammonium chloride (for example, in solution CH ₂ Cl ₂ or THF at a concentration of, e.g., 10 wt.%)).

The wavy lines on the bonds in the fragments of formulas I (5) and I (6) indicate the binding positions of these fragments.

Compounds of formulas XVIA and XVIB can be prepared by reacting a corresponding compound of formula II, as defined above, with a compound of formula XIXA or XIXB

where R ⁴ , Y, X ₁ , X ₂ , X ₃ and X ₄ are as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I.

Compounds of formulas XVIA and XVIB can alternatively be prepared by reacting a corresponding compound of formula IV, as defined above, with a compound of formula XIXC or XIXD

where R ⁴ , X ₁ , X ₂ , X ₃ and X ₄ are as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I.

Compounds of formula XVII can be prepared by reacting the corresponding compound of formula II, as defined above, with a compound of formula XX

where Y and R ^3b are as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I.

Alternatively, compounds of formula XVII can be prepared by reacting a corresponding compound of formula I with a compound corresponding to a compound of formula XIX, in which instead of R ⁷ there is a group R ^b , where R ^b is as defined above, for example, under the conditions described above regarding for preparing compounds of formula Ia.

Compounds of formulas XV and XX may be prepared by reacting a corresponding compound of formula X as defined above, respectively, with a compound of formula XVI as defined above, or with a compound of formula XXI

where R ^3b is as defined above, for example, under conditions similar to the conditions described above for the synthesis of compounds of formula I.

Compounds of the formula XVI, XVIII, XIX, XIXA, XIXB, XIXC, XIXD and XXI are either commercially available, are well known in the literature, or can be obtained either by analogy with the methods described above or using conventional synthesis procedures according to standard procedures from readily available starting materials using appropriate reagents and reaction conditions. For example, compounds of formulas XIXA and XIXB can be prepared by reacting a corresponding compound of formula XIXC or XIXD (as appropriate) with a compound of formula X, for example, under conditions similar to those described above.

The compounds of formulas I and Ia as defined above, and derivatives of both of them are hereinafter referred to as “compounds of the invention”.

Thus, preferred compounds of the invention include those of the examples described below. In this regard, the compounds of the invention that may be mentioned include the following:

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OEt);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-n-Pr);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Pr);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (COO-cyclopentyl);

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Z);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH ₂ -3- (5-Me-isoxazole));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH ₂ -3-pyridine);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Bu);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OEt);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclohexyl);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu);

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH ₂ CH ₂ OPh (3-CF ₃ ));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (4-Cl));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (3-MeO));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (2-Br));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (4-Me));

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-4-heptyl);

Ph (3-Cl) (5-OCHF ₂ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (OMe);

Ph (3-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab;

Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe);

Ph (3-Cl, 5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH);

Ph (3-Cl, 5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OH);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Pro-Pab;

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Pro-Pab (OMe);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((2-amidino) -5-pyridinyl);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((2-methoxyamidino) -5-pyridinyl);

Ph (3-Cl, 5-OCHF ₂ (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((5-amidino) -2-pyrimidinyl);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((5-methoxyamidino) -2-pyrimidinyl);

Ph (3-Cl, 5-OCHF ₂ (R) CH (OH) C (O) -Aze-Pab (3-F);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (OMe);

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) and

Ph (3-Cl, 5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (OMe).

The compounds of the invention may exhibit tautomerism. All their tautomeric forms and mixtures are included in the scope of this invention. Specific tautomeric forms that may be mentioned include those associated with the position of the double bond in the amidine functional group in the compound of formula Ia and with the position of the substituent R ⁵ .

In addition, the compounds of the invention contain one or more asymmetric carbon atoms and therefore may exhibit optical and / or diastereoisomerism. Diastereoisomers can be resolved using conventional techniques, for example, chromatography. Many stereoisomers can be isolated by resolution of a racemic or other mixture of these compounds using conventional techniques, for example, HPLC. Alternatively, the desired optical isomers can be obtained by reacting suitable optically active starting materials under conditions that do not result in racemization or epimerization, or by preparing derivatives, for example, with homochiral acid, followed by resolution of the diastereomeric derivatives by conventional methods (e.g., HPLC, chromatography on silica). All stereoisomers are included within the scope of this invention.

The compounds of the invention in which the fragment

is in S-configuration, are preferred.

Preferred compounds of the invention include those in which a structural fragment

is in the R configuration when R ^a is —OH, or is in the S configuration when R ^a is —CH ₂ OH.

The wavy lines on the bonds in the above two fragments indicate the binding positions of these fragments.

Compounds of the invention that may be mentioned include Ph (3-Cl) (5-OCHF ₂ ) -CH (OH) C (O) -Aze-Pab (where, in this case, Aze means azetidine-2-carboxylate ( that is, in (R) - and / or (S) -conformations)) as well as equivalent compounds in which, instead of a hydrogen atom in the amidine part, in the Pab there is an -OR ⁶ group (as defined above) in which R ⁶ represents C _1-3 alkyl (i.e., Ph (3-Cl) (5-OCHF ₂ ) -CH (OH) C (O) -Aze-Pab (OMe), Ph (3-Cl) (5-OCHF ₂ ) -CH (OH) C (O) -Aze-Pab (OEt), Ph (3-Cl) (5-OCHF ₂ ) -CH (OH) C (O) -Aze-Pab (O-n-Pr) or Ph (3-Cl) (5-OCHF ₂ ) -CH (OH) C (O) -Aze-Pab (O-iso-Pr)). In addition, compounds of the invention that may be mentioned include those that are not specific compounds identified in the previous sentence.

It will be apparent to those skilled in the art that in the methods described above and below, functional groups of intermediates may require protection with protecting groups.

Functional groups that it is desirable to protect include hydroxy, amino and carboxylic acid groups. Suitable hydroxy protecting groups include optionally substituted and / or unsaturated alkyl groups (e.g. methyl, allyl, benzyl or tert-butyl), trialkylsilyl or diarylalkylsilyl groups (e.g. tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyrilyl) and tetra. Suitable protecting groups for carboxylic acids include C _1-6 alkyl or benzyl esters. Suitable protecting groups for amino and amidino include tert-butyloxycarbonyl, benzyloxycarbonyl or 2-trimethylsilylethoxycarbonyl (Teos). Amidine nitrogen can also be protected by hydroxy or alkoxy groups and can be either single or bi-protected.

Protection and deprotection of functional groups may occur before or after combination, or before or after any other reaction in the above schemes.

Protecting groups can be removed in accordance with techniques that are well known to those skilled in the art and as described below.

It will be apparent to those skilled in the art that in order to obtain the compounds of the invention in an alternative and in some cases more convenient way, the specific steps of the methods mentioned above can be performed in a different order and / or specific reactions can be performed at another step in the general process (that is, in accordance with a particular reaction, substituents can be added to various intermediates and / or chemical transformations of various intermediates mentioned above are carried out). This may negate or necessitate the need for protective groups.

The type of chemistry used will dictate the need and type of protective groups, as well as the sequence of the synthesis.

The use of protecting groups is thoroughly described in "Protective Groups in Organic Chemistry", edited by JWF McOmie, Plenum Press (1973), and "Protective Groups in Organic Synthesis", 3 ^nd edition, TWGreene & PGM Wutz, Wiley-lnterscience (1999).

Protected derivatives of the compounds of the invention can be chemically converted to the compounds of the invention using standard deprotection techniques (e.g., hydrogenation). It will also be apparent to those skilled in the art that certain compounds of formula Ia may also be mentioned as being “protected derivatives” of compounds of formula I.

Medical and pharmaceutical applications

The compounds of the invention may have pharmacological activity per se. Compounds of the invention that may possess such activity include, but are not limited to compounds of formula I.

However, other compounds of the invention (including compounds of formula Ia) may not have such activity, but may be administered parenterally or orally and then metabolized in the body to form compounds that are pharmacologically active (including, but not limited to, the corresponding compounds of formula I ) Such compounds (also including compounds that may have some pharmacological activity, but this activity is significantly lower than that of the "active" compounds to which they are metabolized) can therefore be described as "prodrugs" of the active compounds.

Thus, the compounds of the invention are useful because they have pharmacological activity and / or are metabolized in the body after oral or parenteral administration to form compounds that have pharmacological activity. Therefore, the compounds of the invention are indicated as pharmaceuticals.

Thus, according to another aspect of the present invention, there are provided compounds of the invention for use as pharmaceuticals.

In particular, the compounds of the invention are potent thrombin inhibitors either alone and / or (for example, in the case of prodrugs) are metabolized after administration to form potent thrombin inhibitors, for example, as can be demonstrated in the tests described below.

The term “thrombin inhibitor prodrug” includes compounds that form a thrombin inhibitor in an experimentally detectable amount and within a predetermined time (eg, approximately 1 hour) after oral or parenteral administration (see, for example, Test E below) or, alternatively , after incubation in the presence of liver microsomes (see, for example, Test G below).

Thus, it is expected that the compounds of the invention will be useful in conditions where thrombin inhibition is required and / or conditions where anticoagulant therapy is indicated, including the following.

Treatment and / or prevention of thrombosis and hypercoagulation in the blood and / or tissues of animals, including humans

It is known that hypercoagulation can lead to thromboembolic diseases. Conditions associated with hypercoagulation and thromboembolic diseases that may be mentioned include hereditary or acquired resistance to activated protein C, such as a mutation of factor V (Leiden factor V), and hereditary or acquired deficiencies of antithrombin III, protein C, protein S , heparin cofactor II. Other conditions known to be associated with hypercoagulation and thromboembolic disease include circulating antibodies against phospholipids (lupus anticoagulant), heparin-induced homocysteinemia and thrombocytopenia and fibrinolysis disorders, as well as coagulation syndromes, such as disseminated blood vessels, syndrome (DIC)) and vascular damage in general (for example, as a result of surgery).

Treating conditions in which there is an undesirable excess of thrombin without symptoms of hypercoagulation, such as neurodegenerative diseases such as Alzheimer's disease

Specific disease conditions that may be mentioned include therapeutic and / or prophylactic treatment of venous thrombosis (e.g., deep vein thrombosis) and pulmonary embolism, arterial thrombosis (e.g., myocardial infarction, unstable angina pectoris caused by thrombosis, stroke, and peripheral arterial thrombosis) ), and systemic embolism, usually from the atrium during atrial fibrillation (for example, non-valve atrial fibrillation) or from the left ventricle after transmural myocardial infarction, or caused by oh congestive heart failure, prophylaxis of reocclusion (i.e. thrombosis) after thrombolysis, percutaneous transluminal angioplasty (PTA) and coronary bypass surgery, prevention of re-thrombosis after microsurgery and vascular surgery in general.

Additional indications include therapeutic and / or prophylactic treatment of disseminated intravascular coagulation caused by bacteria, multiple trauma, intoxication, or any other mechanism; treatment with anticoagulants when the blood comes in contact with foreign surfaces in the body, such as vascular grafts, vascular stents, vascular catheters, mechanical and biological artificial valves or any other medical device, and treatment with anticoagulants, when the blood comes in contact with medical devices outside the body, such as , during cardiovascular surgery using a cardiopulmonary bypass or hemodialysis, therapeutic and / or prophylactic treatment of idiopathic and respiratory distress syndrome of adults, pulmonary fibrosis after treatment with radiation or chemotherapy, septic shock, septicemia, inflammatory reactions, which include edema, acute or chronic atherosclerosis, such as coronary heart disease and the formation of atherosclerotic plaques, disease cerebral arteries, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral artery disease, ischemia, angina pectoris (including unstable angina pectoris), turnip perfusion injury, restenosis after intraluminal angioplasty (PTA), and coronary artery bypass surgery.

Compounds of the invention that inhibit trypsin and / or thrombin may also be useful in the treatment of pancreatitis.

Thus, the compounds of the invention are indicated for both therapeutic and / or prophylactic treatment of these conditions.

In accordance with another aspect of the present invention, there is provided a method of treating a condition in which thrombin inhibition is required, wherein a subject suffering from or suffering from such a condition is administered a therapeutically effective amount of a compound of the invention.

The compounds of this invention will usually be administered orally, intravenously, subcutaneously, buccally, rectally, through the skin, nasally, into the trachea, in the bronchi, by any other parenteral route or by inhalation, in the form of pharmaceutical preparations containing the compound of the invention in the form of or free base or a pharmaceutically acceptable non-toxic salt of joining an organic or inorganic acid in a pharmaceutically acceptable dosage form.

Preferred routes of administration of the compounds of the invention are oral. Preferred pharmaceutical preparations include modified release pharmaceutical compositions containing the compounds of the invention. The term “modified release” pharmaceutical composition is well understood by those skilled in the art as including any composition for which the onset and / or rate of release of a drug (i.e., a compound of the invention) is altered by galenical procedures, and thus includes the definition provided for in United States Pharmacopeia (USP XXII) on pages xIiii and xIiv of the introduction / introductory part, with a relevant description of this document is incorporated herein by reference.

Thus, suitable modified release formulations may be prepared by one skilled in the art according to standard pharmaceutical techniques (see, for example, Pharmaceutisch Weekblad Scientific Edition, 6, 57 (1984); Medical Applications of Controlled Release, Vol II, eds. Langer and Wise (1984) Bocaration, Florida, on pages 1-34; Industrial Aspects of Pharmaceuticats, ed. Sandel, Swedish Pharmaceutical Press (1993) on pages 93-104; pages 191-211 "Pharmaceutics: The Science of Dosage Form Design", ed. ME Aulton (1988) (Churchill Livingstone)).

Thus, preferred modified release formulations include those in which the corresponding compound of the invention is enclosed in a polymer matrix. In this regard, the authors prefer that preparations comprising the compounds of the invention be proposed for oral administration in the form of a so-called “swelling” modified release system or a “gel-forming matrix” modified release system in which the compound of the invention is proposed together with a polymer that swells in an aqueous medium (that is, with a "hydrophilic gel-forming component").

In particular, the authors prefer that the compounds of the invention be formulated together in a gelling matrix composition comprising an iota-carrageenan and one or more than one neutral gelling polymer.

Iota-carrageenan is preferably present in such a preferred preparation at a level of more than 15% by weight. Preferred types of iota-carrageenan include “pharmaceutical” iota-carrageenan (available from FMC Biopolymer), whose viscosity is at least 5 centipoise (cP), preferably in the range of 5-10 cP (for a 1.5% solution heated up to 82 ° C, after which the viscosity is measured at 75 ° C using a Brookfield LV viscometer equipped with a No. 1 axis moving at a speed of 30 rpm) and a technical iota-carrageenan (available from Fluka Biochemica), whose viscosity is preferably is not less than 14 MPa · s for a 0.3% aqueous solution heated to 20 ° C, after which the viscosity measured by a falling ball viscometer, Haake type used together with thermostat Lauda C3 and Hakke Mess-System III, and using gold-coated stainless steel balls of a density of 7.8 g / cm ^3.

The neutral gelling polymer may be single or a mixture of more than one neutral degradable polymer having gelling properties and having significant pH-independent solubility. The level of neutral gelling polymer in the preparation is preferably more than 10%, but preferably more than 20% by weight.

Suitable neutral gelling polymers include polyethylene oxide (PEO), derivatives and members of the PEO family (e.g., polyethylene glycol (PEG), preferably existing in nature in the solid state, with a suitable molecular weight or viscosity). If PEO is used as a single neutral gelling polymer, then it preferably has an MM (molecular weight) ≥4 million (4M), corresponding to an interval of viscosity of an aqueous solution of 1650-5500 mPa · s (or 1650-5500 cP; measured for 1% aqueous solution at 25 ° C using a Brookfield RVF viscometer with axis No. 2 at 2 rpm). Other examples of suitable PEOs include PEOs with an MM of approximately 5 million (5M) corresponding to an aqueous solution viscosity range of 5500-7500 mPa · s, or PEOs with an MM of approximately 8 million (8M) corresponding to an aqueous solution viscosity range of 10,000-15,000 MPa · from. This range covers the viscosity value for a typical solution (in cP), measured at 25 ° C, guaranteed for this polymer in USP 24 / NF (National Formulary) 19, 2000 edition, pp.2285-2286. If PEG is used as a single neutral gelling polymer, it preferably has a high molecular weight, for example, MM of approximately 20,000, which corresponds to a viscosity range of 2700-3500 mPa · s (or 2700-3500 cP), measured using a 50% aqueous solution (w / w) at 20 ° C. using a capillary viscometer (Ubbelohde or equivalent). [Ref: European Pharmacopoeia 3 ^rd Ed., 2000, Supplement, pp.908-909.]

Other suitable gelling polymers include cellulose derivatives such as hydroxypropyl methyl cellulose (HPMC) or hydroxyethyl cellulose (HEC) with suitable high viscosities (for example, "HPMC 10,000 cP", "HPMC 15,000 cP", "HEC, type HH" or " HEC type H "). If hydroxypropylmethyl cellulose polymers such as HPMC 10,000 cP and HPMC 15,000 cP are used as a single neutral polymer, they have an apparent viscosity of 7500-14000 mPa · s (or 7500-14000 cP) and 11250-21000 mPa · respectively s (or 11250-21000 cP) when measured at 20 ° C using a 2% (w / w) aqueous solution, calculated relative to the dry matter, using a capillary viscometer (Ubbelohde or equivalent). One type of hydroxyethyl cellulose polymer, for example, Natrosol 250 Pharma, type HH, from Hercules Incorporated (Aqualon), typically shows a Brookfield viscosity of approximately 20,000 mPa · s when using a Brookfield Synchro-Lectric Model LVF at 1% concentration solution, with axis No. 4, axis speed of 30 rpm, with a factor of 200, 25 ° C (see Natrosol Physical and Chemical Properties booklet, 33.007-E6 (1993), p.21).

Specific preparations that may be mentioned include those in which the compound of the invention is prepared in conjunction with iota-carrageenan and HPMC (10,000 cP) in a ratio of 50:50 (wt%) or together with iota-carrageenan and REO 4M in a ratio of 50:50 (wt.%).

Preferred additional excipients in such formulations include lubricants, such as sodium fumarate.

Depending on the disorder and the patient who needs treatment and the method of administration, these compositions may be administered in various doses.

In addition, the compounds of the invention can be combined and / or co-administered with antithrombotic agent (s) with another mechanism of action, such as one or more of the following: with antiplatelet agents, acetylsalicylic acid, ticlopidine and clopidogrel; thromboxane receptor and / or synthetase inhibitors; fibrinogen receptor antagonists; prostacyclin mimetics; phosphodiesterase inhibitors; antagonists of ADP (adenosine diphosphate) receptors (P ₂ T) and carboxypeptidase U inhibitors (CPU).

The compounds of the invention can be further combined and / or co-administered with thrombolytic agents, such as one or more tissue plasminogen activators (natural, recombinant or altered), streptokinase, urokinase, prourokinase, anisoylated plasminogen-streptokinase activator complex (APSAC), plasminogen activators of the salivary glands of animals and the like, in the treatment of thrombotic diseases, in particular myocardial infarction.

Thus, according to another aspect of the present invention, there is provided a pharmaceutical preparation comprising a compound of the invention in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

Suitable daily doses of the compounds of the invention for the therapeutic treatment of humans are approximately 0.001-100 mg / kg body weight for oral administration and 0.001-50 mg / kg body weight for parenteral administration.

For the avoidance of doubt, the term “treatment,” as used herein, includes therapeutic and / or prophylactic treatment.

The compounds of this invention have the advantage over the compounds known from the prior art in that they can be more effective, less toxic, have a longer duration of action, have a wider range of activity, be more potent, have fewer side effects, more easily absorbed and / or have a better pharmacokinetic profile (for example, higher oral bioavailability and / or lower clearance), and / or have other useful pharmacological, physical or chemical skie properties. The compounds of the invention may have an additional advantage over prior art compounds in that they can be administered less frequently.

Biological tests

The following analysis procedures may be used.

Test a

Determination of coagulation time of blood plasma after the addition of thrombin (thrombin time) (TT)

The inhibitor solution (25 μl) was incubated with plasma (25 μl) for 3 minutes. Then add human thrombin (T 6769, Sigma Chem. Co or Hematologic Technologies) in a buffer solution, pH 7.4 (25 μl, 4.0 NIH (National Institute of Health) units / ml), and clotting time is measured on an automatic device ( COP 10, Amelung).

Thrombin time (TT) is expressed in absolute values (seconds), as well as in the ratio of TT without inhibitor (TT ₀ ) to TT with inhibitor (TT _i ). The dependence curves of the last relations (1-0 interval) on the inhibitor concentration (in the log transformation) are built, and they correspond to the sigmoidal dose-response curves according to the equation

y = a / [1+ (x / IC ₅₀ ) ^S ],

where a = maximum interval, i.e. 1; s = slope of the dose-response curve and IC ₅₀ = inhibitor concentration that doubles the clotting time. The calculations are performed on a personal computer (PC) using GraFit Version 3 software, setting the equation to 0 first, determine the final value = 1 (Erithacus Software, Robin Leatherbarrow, Imperial College of Science, London, United Kingdom of Great Britain and Northern Ireland).

Test b

Determination of thrombin inhibition using an automated chromogenic assay

The efficacy of a thrombin inhibitor is measured by the method using a chromogenic substrate on a Plato 3300 automated microplate processor (Rosys AG, CH-8634 Hombrechtikon, Switzerland) using half-well 96-well microtiter microtiter plates (Costar, Cambridge, MA, USA; 3690) . Stock solutions of the test substance in DMSO (72 μl), 0.1-1 mmol / L, are subsequently diluted with DMSO as 1: 3 (24 + 48 μl) to obtain ten different concentrations, which are analyzed as samples for analysis. 2 μl of the test sample is diluted with 124 μl of assay buffer, 12 μl of a chromogenic substrate solution (S-2366, Chromogenix, Mölndal, Sweden) in the assay buffer, and 12 μl of a solution of α-thrombin (human α-thrombin, Sigma Chemical Co . or Hematologic Technologies) in an assay buffer and the samples are mixed. The final analyzed concentrations are: test substance 0.00068-13.3 μmol / L, S-2366 0.30 mmol / L, α-thrombin 0.020 NIH units / ml. To calculate the percent inhibition for the test samples, a linear increase in absorption was used during 40 minutes of incubation at 37 ° C compared with control samples without an inhibitor. An automatically recorded IC ₅₀ value corresponding to the concentration of the inhibitor causing 50% inhibition of thrombin activity is calculated from the curve of the log of the concentration versus% inhibition.

Test c

Determination of the inhibition constant K _i for human thrombin

The K _i determinations are made using the method using a chromogenic substrate at 37 ° C on a Cobas Bio centrifuge analyzer (Roche, Basel, Switzerland). The residual enzymatic activity after incubation of human α-thrombin with a test compound taken in various concentrations is determined at three different substrate concentrations and measured as a change in optical density at 405 nm.

Solutions of the test compound (100 μl; usually in a buffer or physiological saline containing 10 g / L BSA) are mixed with 200 μl of human α-thrombin (Sigma Chemical Co) in assay buffer (0.05 mol / L Tris-HCl, pH 7.4, with an ionic strength of 0.15, adjusted with NaCl) containing BSA (10 g / L), and analyzed as samples on a Cobas Bio. To 320 μl of S-2238 substrate (Chromogenix AB, Mölndal, Sweden), 60 μl of the sample together with 20 μl of water are added to the assay buffer and the density change (ΔA / min) is recorded. Final concentrations of S-2238 are 16, 24 and 50 μmol / L, and thrombin 0.125 units NIH / ml.

The reaction rate in the stationary state is used to plot Dixon graphs, that is, diagrams showing the change in 1 / (ΔA / min) versus inhibitor concentration. For reversible competitive inhibitors, the experimental points for various substrate concentrations usually form straight lines that intersect at x = -K _i .

Test d

Determination of Partially Activated Thromboplastin Time (ARTT)

ARTT is determined in pooled samples of normal human citrate plasma using the PTT Automated 5 reagent manufactured by Stago. Inhibitors are added to plasma (10 μl of inhibitor solution to 90 μl of plasma), incubated with ARTT reagent for 3 minutes, followed by the addition of 100 μl of calcium chloride solution (0.025 M) and ARTT is determined using a K10 coagulation analyzer (Amelung) in accordance with Reagent manufacturer instructions.

Coagulation time is expressed in absolute values (seconds), as well as in the ratio of ARTT without inhibitor (ARTT ₀ ) to ARTT with inhibitor (APTT _i ). The dependence curves of the last relations (1-0 interval) on the inhibitor concentration (in the log transformation) are built, and they correspond to the sigmoidal dose-response curves according to the equation

y = a / [1+ (x / IC ₅₀ ) ^s ],

where a = maximum interval, i.e. 1; s = slope of the dose-response curve and IC ₅₀ = inhibitor concentration that doubles the clotting time. The calculations are performed on a PC using GraFit Version 3 software, setting the equation to 0 at first, determining the final value = 1 (Erithacus Software, Robin Leatherbarrow, Imperial College of Science, London, United Kingdom of Great Britain and Northern Ireland).

IC ₅₀ APTT is defined as the concentration of inhibitor in human plasma that doubled the partial activated thromboplastin time.

Test e

Ex vivo thrombin time assay

Thrombin inhibition after oral or parenteral administration of the compounds of the invention dissolved in ethanol / Solutol K / water (5: 5: 90) is tested in conscious rats who are given a catheter for sampling one or two days before the experiment. blood from the carotid artery. On the day of the experiment, at a fixed time after the administration of the compound, blood samples are taken in plastic tubes containing 1 part sodium citrate solution (0.13 mol per 1 liter) and 9 parts blood. The tubes are centrifuged to obtain platelet-poor plasma.

Plasma samples in a volume of 50 μl precipitated with 100 μl of cooled acetonitrile. Samples are centrifuged for 10 minutes at 4000 rpm. 75 μl of the supernatant is diluted with 75 μl of 0.2% formic acid. The resulting solutions in a volume of 10 μl are analyzed using LC-MS / MS and, using calibration curves, the concentrations of the thrombin inhibitor are determined.

Test f

Determination of plasma clearance in rats

Plasma clearance is evaluated in male Sprague Dawley rats. The compound is dissolved in water and administered as a subcutaneous bolus injection at a dose of 4 μmol / kg. Blood samples are collected at repeated intervals up to 5 hours after drug administration. Blood samples are centrifuged, the plasma is separated from blood cells and transferred to tubes containing citrate (at a final concentration of 10%). Plasma samples in a volume of 50 μl precipitated with 100 μl of cooled acetonitrile. Samples are centrifuged for 10 minutes at 4000 rpm. 75 μl of the supernatant is diluted with 75 μl of 0.2% formic acid. The resulting solutions in a volume of 10 μl are analyzed using LC-MS / MS and, using calibration curves, the concentrations of the thrombin inhibitor are determined. The area under the plasma concentration-time curve is estimated using the log / linear trapezoidal formula and extrapolated to infinite time. The plasma clearance (CL) of the compound is then determined as

CL = dose / AUC (area under the curve).

Values are expressed in ml / min / kg.

Test g

In vitro stability determination

Liver microsomes are obtained from liver samples of Sprague-Dawley rats and humans according to internal SOP (standard operating procedures). Compounds are incubated at 37 ° C with a total concentration of microsomal protein of 3 mg / ml in 0.05 mol / L Tris buffer at pH 7.4 in the presence of cofactors NADH (2.5 mmol / L) and NADPH (0, 8 mmol / l). The initial concentration of the compound is 5 or 10 μmol / L. Samples were taken for analysis up to 60 minutes after the start of incubation. Enzymatic activity in the selected sample is immediately stopped by the addition of 20% muristinic acid in a volume corresponding to 3.3% of the total volume of the sample. The concentration of the compound remaining in the sample after 60 minutes (FINAL CONC) is determined by LCMS using the sample collected at time zero as a reference sample (START CONC).% Of the destroyed thrombin inhibitor is calculated as

100% × {[START CONC] - [FINAL CONC]} / [START CONC].

Test n

Arterial thrombosis model

Vascular damage is induced by application of iron chloride (FeCl ₃ ) locally to the carotid artery. Rats were anesthetized with an intraperitoneal injection of sodium pentobarbital (80 mg / kg; Apoteksbolaget; Umeå, Sweden) followed by continuous infusion (12 mg / kg / h) for the duration of the experiment. The body temperature of the rat is maintained at 38 ° C during the experiment using external heating. The experiment begins with a 5-minute control period. Five minutes later, human ¹²⁵ I-fibrinogen (80 kBq; IM53; Amersham International, Buckinghamshire, United Kingdom of Great Britain and Northern Ireland) was intravenously administered and used as a marker for the subsequent incorporation of fibrin (ogen) into the thrombus. The proximal end of the carotid artery segment is placed in a plastic tube (6 mm; Silastic®, Dow Corning, MI, USA), open along containing impregnated FeCl ₃ (2 μl; 55 wt.%; Merck, Darmstadt, Germany) filter paper (diameter 3 mm; 1F; Munktell, Grycksbo, Sweden). The left carotid artery is exposed to FeCl ₃ for 10 minutes and then removed from the plastic tube and soaked in saline. Fifty minutes later, the carotid artery is removed and washed in physiological saline. Reference blood samples were also taken to determine ¹²⁵ I-activity of blood, 10 minutes after injection of ¹²⁵ I-fibrinogen and at the end of the experiment. ¹²⁵ I-activity in reference blood samples and a segment of a vessel is measured in a gamma counter (1282 Compugamma; LKB Wallac Oy, Turku, Finland) on the day of the experiment. The thrombus size is defined as the amount of ¹²⁵ I-activity included in the segment of the vessel, relative to ¹²⁵ I-activity in the blood (imp / min / mg).

General experimental techniques

TLC was performed on silica gel. Chiral HPLC analysis was performed using a 46 mm x 250 mm Chiralcel OD column with a 5 cm pre-column. The column temperature was maintained at 35 ° C. A flow rate of 1.0 ml / min was used. A Gilson 115 UV detector was used at 228 nm. The mobile phase consisted of hexanes, ethanol and trifluoroacetic acid, and the corresponding ratios were given for each compound. Typically, the product was dissolved in a minimal amount of ethanol and diluted with the mobile phase.

LC-MS / MS was performed using an HP-1100 instrument equipped with a CTC-PAL injector and a ThermoQuest column, Hypersil BDS-C18, 5 μm, 4 × 100 mm. Used the MS-detector API-3000 (Sciex). The flow rate was 1.2 ml / min, and the mobile phase (gradient) consisted of 10-90% acetonitrile with 90-10% 4 mm aqueous ammonium acetate, each of which contained 0.2% formic acid .

¹ H NMR spectra were recorded using tetramethylsilane as the internal standard. ¹³ C NMR spectra were recorded using nomenclature deuterated solvents as an internal standard.

Example 1

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu)

(1) 3-Chloro-5-methoxybenzaldehyde

3,5-Dichloroanisole (74.0 g; 419 mmol) in THF (200 ml) was added dropwise to magnesium metal (14.2 g; 585 mmol; pre-washed with 0.5 N HCl) in THF (100 ml) at 25 ° C. After this addition, 1,2-dibromethane (3.9 g; 20.8 mmol) was added dropwise. The resulting dark brown mixture was refluxed for 3 hours. The mixture was cooled to 0 ° C. and N, N-dimethylformamide (60 ml) was added in one portion. The mixture was distributed using diethyl ether (3 × 400 ml) and 6 N. HCl (500 ml). The combined organic extracts were washed with brine (300 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give an oil. Flash chromatography (2x) on silica gel eluting with a Hex / EtOAc mixture (4: 1) gave the subtitled compound (38.9 g; 54%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.90 (s, 1H), 7.53 (s, 1H), 7.38 (s, 1H,), 7.15 (s, 1H), 3.87 (s, 3H).

(2) 3-Chloro-5-hydroxybenzaldehyde

A solution of 3-chloro-5-methoxybenzaldehyde (22.8 g; 134 mmol; see step (1) above) in CH ₂ Cl ₂ (250 ml) was cooled to 0 ° C. Boron tribromide (15.8 ml; 167 mmol) was added dropwise over 15 minutes. After stirring the reaction mixture for 2 hours, H ₂ O (50 ml) was slowly added. Then the solution was extracted with Et ₂ O (2 × 100 ml). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (4: 1) gave the subtitled compound (5.2 g; 25%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.85 (s, 1H), 7.35 (s, 1H), 7.20 (s, 1H), 7.10 (s, 1H), 3.68 (s, 1H).

(3) 3-Chloro-5-difluoromethoxybenzaldehyde

A solution of 3-chloro-5-hydroxybenzaldehyde (7.5 g; 48 mmol; see step (2) above) in 2-propanol (250 ml) and 30% KOH (100 ml) was heated to reflux. The reaction mixture was bubbled with CHCIF ₂ with stirring for 2 hours. The reaction mixture was cooled, acidified with 1 N. HCl and extracted with EtOAc (2 × 100 ml). The organic extracts were washed with brine (100 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (4: 1) gave the subtitled compound (4.6 g; 46%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.95 (s, 1H), 7.72 (s, 1H), 7.52 (s, 1H), 7.40 (s, 1H), 6.60 (t, J _HF = 71.1 Hz, 1H )

(4) Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OTMS) CN

A solution of 3-chloro-5-difluoromethoxybenzaldehyde (4.6 g; 22.3 mmol; see step (3) above) in CH ₂ Cl ₂ (200 ml) was cooled to 0 ° C. Znl ₂ (1.8 g; 5.6 mmol) and trimethylsilyl cyanide (2.8 g; 27.9 mmol) were added, the reaction mixture was allowed to warm to room temperature and stirred for 15 hours. This mixture was partially concentrated in vacuo to give the subtitled compound in the form of a liquid, which was used directly in the step (5) below without further purification or characterization.

(5) Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (NH) OEt

Ph (3-Cl) (5-OCHF ₂ - (R, S) CH (OTMS) CN (6.82 g; presumably 22.3 mmol; see step (4) above) was added dropwise to the HCl / EtOH mixture ( 500 ml) The reaction mixture was stirred for 15 hours, then partially concentrated in vacuo to give the subtitle compound as a liquid, which was used in step (6) below without further purification or characterization.

(6) Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (O) OEt

Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (NH) OEt (6.24 g; presumably 22.3 mmol; see step (5) above) was dissolved in THF ( 250 ml), 0.5 M H ₂ SO ₄ (400 ml) was added and the reaction mixture was stirred at 40 ° C for 65 hours, cooled and then partially concentrated in vacuo to remove most of the THF. The reaction mixture was then extracted with Et ₂ O (3 × 100 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to obtain the subtitle compound as a solid, which was used in step (7) without further purification or determination characteristics.

(7) Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (O) OH

A solution of Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (O) OEt (6.25 g; presumably 22.3 mmol; see step (6) above) in 2- propanol (175 ml) and 20% KOH (350 ml) were stirred at room temperature for 15 hours. The reaction mixture was then partially concentrated in vacuo to remove most of the 2-propanol. The remaining mixture was acidified with 1 M H ₂ SO ₄ , extracted with Et ₂ O (3 × 100 ml), dried (Na ₂ SO ₄ ) and concentrated in vacuo to give a solid. Flash chromatography on silica gel eluting with a mixture of CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1) gave the ammonium salt of the subtitle compound. This ammonium salt was then dissolved in a mixture of EtOAc (75 ml) and H ₂ O (75 ml) and acidified with 2 N HCl. The organic layer was separated and washed with brine (50 ml), dried (Na ₂ SO ₄ ) and concentrated in vacuo to give the subtitle compound (3.2 g; 57% from steps (4) to (7)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, J _HF = 71.1 Hz, 1H), 5.16 (s, 1H).

(8) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (a) and Ph (3-Cl) (5-OCHF ₂ ) - (S) CH ( OAc) C (O) OH (b)

Mixture of Ph (3-Cl) (5-OCHF ₂ ) - (R, S) CH (OH) C (O) OH (3.2 g; 12.7 mmol; see step (7) above) and Lipases PS " Amano "(~ 2.0 g) in vinyl acetate (125 ml) and MTBE (125 ml) was heated under reflux for 48 hours. The reaction mixture was cooled, filtered through Celite® and the filter cake was washed with EtOAc. The filtrate was concentrated in vacuo and flash chromatographed on silica gel eluting with a mixture of CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1) to give the ammonium salt of the subtitled compounds (a) and (b). Compound (a) as a salt was dissolved in H ₂ O, acidified with 2 N HCl and extracted with EtOAc. The organic layer was washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitle compound (a) (1.2 g; 37%).

For the subtitle compound (a)

¹ H NMR (300 MHz; CD ₃ OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, J _HF = 71.1 Hz, 1H), 5.17 (s, 1H).

(9) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (1.1 g; 4.4 mmol; see step (8) above) and H-Aze- Pab (Teoc) (see international patent application WO 00/42059; 2.6 g; 5.7 mmol) in DMF (50 ml) at 0 ° C, RuBOP (2.8 g; 5.3 mmol) and collidine ( 1.3 g; 10.6 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature for an additional 15 hours. The reaction mixture was concentrated in vacuo and flash chromatographed on silica gel (3x), eluting first with CHCl ₃ / EtOH (9: 1) then EtOAc / EtOH (20: 1) and finally eluting with a mixture of CH ₂ Cl ₂ / CH ₃ OH (95: 5) to obtain the subtitled compound (1.0 g; 37%) as a white solid.

¹ H NMR (300 MHz; CD ₃ OD; mixture of rotamers) δ 7.79-7.85 (d, J = 8.7 Hz, 2H), 7.15-7.48 (m, 5H), 6.89 and 6.91 (t, j _HF = 71.1 Hz, 1H), 5.12 and 5.20 (s, 1H), 4.75-4.85 (m, 1H), 3.97-4.55 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.15 (m, 2H), 0.09 ( s, 9H).

MS (m / z) 611 (M + 1) ⁺ .

(10) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu, Teoc)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.051 g; 0.08 mmol; see step (9) above) was dissolved in 3 ml of acetonitrile and 0.062 g (0.5 mmol) of O-cyclobutylhydroxylamine hydrochloride was added. The mixture was heated at 70 ° C. for 4.5 hours. The solvent was evaporated and the residue was partitioned between water and ethyl acetate. The aqueous phase was extracted two more times with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na ₂ SO ₄ ), filtered and evaporated. Yield 0.054 g (95%).

¹ H-NMR (400 MHz; CD ₃ OD): δ 8.66-8.50 (m, 1H), 7.45 (d, 2H), 7.29 (m, 3H), 7.15 (m, 2H), 6.88 (t, 1H basic rotamer), 6.85 (t, 1H secondary rotamer), 5.18 (s, 1H secondary rotamer), 5.12 (s, 1H secondary rotamer), 5.16 (m, 1H secondary rotamer), 4.78 (m, 1H main rotamer), 4.70 ( m, 1H), 4.50-4.30 (m, 3H), 4.19-3.93 (m, 3H), 2.71-2.44 (m, 1H), 2.34-2.11 (m, 5H), 1.78 (m, 1H), 1.62 ( m, 1H), 0.96 (m, 2H), 0.01 (s, 9H).

(11) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu, Teoc) (0.054 g; 0.08 mmol; see step (10) ) above) was dissolved in 0.5 ml of CH ₂ Cl ₂ and 3 ml of TFA. The reaction was allowed to proceed for 60 minutes. TFA was evaporated and the residue was purified using preparative HPLC. Fractions of interest were combined and freeze dried (2x) to give 23 mg (54%) of the title compound.

MS (m / z) 536 (M-1) ^- ; 538 (M + 1) ⁺ .

¹ H-NMR (400 MHz; CD ₃ OD): δ 7.56 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H main rotamer), 6.86 (t, 1H minor rotamer), 5.18 (s, 1H main rotamer, and m, 1H secondary rotamer), 5.11 (s, 1H secondary rotamer), 4.77 (m, 1H main rotamer), 4.58 (m, 1H), 4.42 (m, 2H) , 4.34 (m, 1H main rotamer), 4.15 (m, 1H main rotamer), 4.06 (m, 1H secondary rotamer), 3.97 (m, 1H secondary rotamer), 2.66 (m, 1H secondary rotamer), 2.52 (m, 1H main rotamer), 2.33-2.25 (m, 3H), 2.01-2.20 (m, 2H), 1.75 (m, 1H), 1.59 (m, 1H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.4, 172.3, 171.9, 171.4, 152.3.

Example 2

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH, Teoc)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.148 g; 0.24 mmol; see Example 1 (9) above) was dissolved in 9 ml of acetonitrile and 0.101 g (1.45 mmol) of hydroxylamine hydrochloride was added. The mixture was heated at 70 ° C for 2.5 hours, filtered through Celite® and evaporated. The crude product (0.145 g; 75% purity) was directly used in the next step without further purification.

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH, Teoc) (0.145 g; 0.23 mmol; see step (1) above) was dissolved in 0.5 ml of CH ₂ Cl ₂ and 9 ml of TFA. Reactions were allowed to proceed for 60 minutes. TFA was evaporated and the residue was purified using preparative HPLC. Fractions of interest were collected and freeze-dried (2x) to give 72 mg (62% yield in two steps) of the title compound.

MS (m / z) 482 (M-1) ^- ; 484 (M + 1) ⁺ .

¹ H-NMR (400 MHz; CD ₃ OD): δ 7.58 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H main rotamer), 6.86 (t, 1H minor rotamer), 5.18 (s, 1H main rotamer, and m, 1H secondary rotamer), 5.12 (s, 1H secondary rotamer), 4.77 (m, 1H main rotamer), 4.42 (m, 2H), 4.34 (m, 1H main rotamer), 4.14 (m, 1H secondary rotamer), 4.06 (m, 1H secondary rotamer), 3.95 (m, 1H secondary rotamer), 2.66 (m, 1H secondary rotamer), 2.50 (m, 1H primary rotamer), 2.27 ( m, 1H primary rotamer), 2.14 (m, 1H secondary rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.4, 172.3, 172.0, 171.4, 152.3, 152.1.

Example 3

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.045 g; 0.074 mmol; see Example 1 (9) above) was dissolved in 3 ml TFA and allowed to proceed for 1 h. TFA was evaporated and the residue was dried by freezing from a mixture of water / acetonitrile to obtain 0.043 g (100%) of the title compound as its TFA salt.

¹ H-NMR (400 MHz; CD ₃ OD) rotamers: δ 7.8-7.75 (m, 2H), 7.55-7.5 (m, 2H), 7.35 (m, 1H, main rotamer), 7.31 (m, 1H, minor rotamer), 7.19 (m, 1H, primary rotamer), 7.15 (m, 1H), 7.12 (m, 1H, secondary rotamer), 6.89 (t, 1H, primary rotamer), 6.87 (t, 1H, secondary rotamer), 5.22 (m, 1H, secondary rotamer), 5.20 (s, 1H, primary rotamer), 5.13 (s, 1H, secondary rotamer), 4.80 (m, 1H, primary rotamer), 4.6-4.4 (m, 2H), 4.37 (m, 1H, primary rotamer), 4.19 (m, 1H, primary rotamer), 4.07 (m, 1H, secondary rotamer), 3.98 (m, 1H, secondary rotamer), 2.70 (m, 1H, secondary rotamer), 2.55 (m, 1H, main rotamer), 2.29 (m, 1H, main rotamer), 2.15 (m, 1H, secondary rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD): (carbonyl and / or amidine carbons, rotamers) δ 172.6, 172.5, 172.0, 171.7, 167.0.

MS (m / z) 465 (M-1) ^- ; 467 (M + 1) ⁺ .

Example 4

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (COO-cyclopentyl)

To a solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) (OH) C (O) -Aze-Pab × TFA (74 mg; 0.13 mmol; see Example 3 above) and cyclopentyl chloroformate (44 mg ; 0.30 mmol) in methylene chloride (5 ml) was added aqueous NaOH (0.5 ml; 2 M; 1 mmol). The mixture was stirred at room temperature and the reaction was monitored by HPLC. After 2.5 hours, water was added and the liquid phases were separated. The aqueous phase was extracted twice with methylene chloride. The combined organic phases were dried (MgSO ₄ ) and purified on silica gel (first methylene chloride, then EtOAc). After removing the solvents in vacuo, the solid residue was dissolved in water / acetonitrile and freeze dried to give the title compound as a white solid. Yield 33 mg (44%).

MS (m / z) 579 (M + 1) ⁺ .

¹ H NMR (400 MHz; CD ₃ OD): δ 7.79 (d, 2H), 7.43-7.30 (t, 5H), 7.20-7.11 (t, 2H), 6.90 (t, 1H, main rotamer), 6.87 ( t, 1H, secondary rotamer), 5.19 (dd, 1H, secondary rotamer), 5.18 (s, 1H, primary rotamer), 5.13 (m, 1H), 5.11 (s, 1H, secondary rotamer), 4.78 (dd, 1H , main rotamer), 4.45 (m, 2H), 4.35 (m, 1H, main rotamer), 4.16 (s, 1H, main rotamer), 4.06 (s, 1H, secondary rotamer), 3.97 (s, 1H, secondary rotamer ), 2.68 (m, 1H, secondary rotamer), 2.52 (s, 1H, primary rotamer), 2.28 (s, 1H, primary rotamer), 2.16 (s, 1H, secondary rotamer), 1.90 (m, 2H), 1.77 (m, 4H), 1.61 (m, 2H).

¹³ C NMR (carbonyl and / or amidine protons; 100 MHz): δ 173.6, 173.1, 172.6, 170.3, 165.6.

Example 5

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Z)

The title compound was prepared according to the procedure described in Example 4 above, starting with Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA (73 mg; 0.13 mmol; see example 3 above) and benzyl chloroformate (35 mg; 0.21 mmol). Further purification is required using reverse phase HPLC (0.1 M ammonium acetate / MeCN (40:60)). The appropriate fractions were concentrated in vacuo and extracted with EtOAc. Yield 24 mg (32%).

MS (m / z) 602 (M + 1) ⁺ .

¹ H NMR (400 MHz; CD ₃ OD): δ 7.80 (d, 2H), 7.43-7.25 (m, 8H), 7.20-7.10 (m, 2H), 6.90 (t, 1H, main rotamer), 6.88 ( t, 1H, secondary rotamer), 5.18 (dd, 1H, secondary rotamer), 5.18 (s, 1H, rotamer), 5.11 (s, 1H, rotamer), 4.78 (dd, 1H, main rotamer), 4.45 (m, 2H), 4.34 (m, 1H, main rotamer), 4.15 (s, 1H, main rotamer), 4.06 (s, 1H, secondary rotamer), 3.97 (s, 1H, secondary rotamer), 2.66 (m, 1H, secondary rotamer), 2.51 (s, 1H, primary rotamer), 2.27 (s, 1H, primary rotamer), 2.15 (s, 1H, secondary rotamer).

¹³ C NMR (carbonyl and / or amidine protons; 100 MHz): δ 173.6, 173.1, 172.6, 170.5, 164.9.

Example 6

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 2-Nitro-5-trifluoromethoxybenzoic acid

To a solution of 3-trifluoromethoxybenzoic acid (49.0 g; 0.24 mol) in sulfuric acid (500 ml) at a temperature of less than 0 ° C (ice-MeOH bath) was added a solution of potassium nitrate (31.3 g; 0.31 mol ) in sulfuric acid (200 ml) for 20 minutes. The resulting solution was stirred at 0 ° C. for 2 hours, then warmed to room temperature and stirred for 18 hours. The reaction mixture was poured onto ice and the resulting acid solution was extracted with EtOAc (5x). The combined organic extracts were washed with H ₂ O (1x), brine (2x), H ₂ O (1x) and brine (1x), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (65.7 g) as a solid mixed with HOAc. The crude subtitled compound was dissolved in EtOAc and toluene and concentrated in vacuo to give a HOAc-free solid (58.4 g; 97%), which was used in the next step without further purification.

¹ H NMR (300 MHz; CDCl ₃ ): δ 10.10 (br s, 1H), 8.02 (d, 1H, J = 8 Hz), 7.69 (d, 1H, J = 2 Hz), 7.54 (dd, 1H, J = 2 Hz, J = 8 Hz).

(2) 2-amino-5-trifluoromethoxybenzoic acid

To a solution of 2-nitro-5-trifluoromethoxybenzoic acid (56.8 g; 0.23 mol; see step (1) above) in EtOH (1000 ml) was added 10% Pd / C (5.7 g). A stream of H _{2 was} passed through the resulting solution for 5 hours, filtered through Celite® and concentrated in vacuo to give the crude subtitled compound (49.7 g; 98%) as a solid, which was used in the next step without further purification.

¹ H NMR (300 MHz; CD ₃ OD): δ 7.66 (m, 1H), 7.17 (d, 1H, J = 8 Hz), 6.77 (d, 1H, J = 8 Hz).

(3) 2-amino-3-chloro-5-trifluoromethoxybenzoic acid

Sulfuryl chloride (41.8 g; 0.31 mol) was slowly added to a solution of 2-amino-5-trifluoromethoxybenzoic acid (49.0 g; 0.22 mol; see step (2) above) in HOAc (1200 ml). Gas evolution was observed. The resulting heterogeneous mixture was stirred at room temperature for 1 h. An additional amount of HOAc (300 ml) was added to facilitate mixing, followed by the addition of sulfuryl chloride in 5 ml portions until the starting material was consumed (according to TLC analysis) . The reaction mixture was concentrated in vacuo to give solids, which were washed on an EtOAc rotary evaporator (2x) to remove HOAc, followed by washing with Et ₂ O (1x). The resulting solids were further dried to obtain the HCl salt of the crude subtitled compound (60.5 g; 94%), which was used in the next step without further purification.

¹ H NMR (300 MHz; CD ₃ OD): δ 7.72 (s, 1H), 7.44 (s, 1H), 7.22 (s, interchangeable).

(4) 3-Chloro-5-trifluoromethoxybenzoic acid

To a solution of 2-amino-3-chloro-5-trifluoromethoxybenzoic acid (60.5 g; presumably 0.22 mol; see step (3) above) in 1,4-dioxane (1000 ml) was added 6N. HCl (750 ml). A certain amount of organic matter in the form of oil fell out of solution. The dioxane solution was cooled to a temperature of less than 0 ° C (ice-Meon bath). A solution of sodium nitrite (18.2 g; 0.26 mol) in H ₂ O (250 ml) was added over a dropping funnel over a period of 15 minutes. The resulting solution was stirred for 45 minutes. Phosphoric acid (221.5 ml of 50 wt% in H ₂ O; 291.2 g; 2.20 mol) was slowly added via a dropping funnel. The solution was stirred at 0 ° C. for 1.5 hours, then warmed to room temperature (gas evolution was observed) and stirred for 18 hours. The crude solution was transferred to a separatory funnel and extracted with Et ₂ O (4x). The combined organic extracts were extracted with aqueous NaHCO ₃ (3x). The main aqueous layer was carefully acidified with 6 N HCl and was extracted with CH ₂ Cl ₂ (3x). The CH ₂ Cl ₂ extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (26.5 g; 46% based on 3-trifluoromethoxybenzoic acid) as a solid, which was used in the following stage without further purification.

¹ H NMR (300 MHz; CD ₃ OD): δ 7.98 (s, 1H), 7.83 (s, 1H), 7.58 (s, 1H).

(5) 3-Chloro-5-trifluoromethoxybenzyl alcohol

To a solution of 3-chloro-5-trifluoromethoxybenzoic acid (22.5 g; 93.5 mmol; see step (4) above) in anhydrous THF (1200 ml) in a N ₂ atmosphere at room temperature was added a solution of the BH ₃ · THF complex ( 140 ml 1 M in THF; 140.3 mmol). The solution was refluxed for 2 hours, cooled to room temperature and stirred for 18 hours, carefully quenched with H ₂ O and concentrated in vacuo to remove as much THF as possible. The residue was diluted with EtOAc and the organic phase was washed with brine (3x), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (21.2 g; 100%) as an oil, which was used without further cleaning up.

¹ H NMR (300 MHz; CDCl ₃ ): δ 7.33 (s, 1H), 7.17 (s, 1H), 7.14 (s, 1H), 4.72 (s, 2H), 2.05 (br s, 1H).

(6) 3-Chloro-5-trifluoromethoxybenzaldehyde

A solution of DMSO (16.1 g; 205.9 mmol) in anhydrous CH ₂ Cl ₂ (300 ml) was cooled to -78 ° C. Oxalyl chloride (13.1 g; 103.0 mmol) was added slowly via syringe (gas evolution was observed). The resulting solution was stirred at -78 ° C for 15 minutes. A solution of 3-chloro-5-trifluoromethoxybenzyl alcohol (21.2 g; 93.6 mmol; see step (5) above) in CH ₂ Cl ₂ (200 ml) was added over a dropping funnel over a period of 15 minutes. The opaque solution was stirred at -78 ° C for 40 minutes and DIPEA (60.5 g; 468.0 mmol) was added over a dropping funnel over 10 minutes. The resulting homogeneous solution was stirred at -78 ° C for 1.5 hours, then warmed to room temperature and stirred for 18 hours. The crude solution was concentrated in vacuo, the residue was diluted with EtOAc and washed with H ₂ O (1x), 2 N. HCl (1x), brine (1x), aqueous NaHCO ₃ (1x) and brine (1x). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (19.9 g; 95%), which was used in the next step without further purification.

¹ H NMR (300 MHz; CDCl ₃ ): δ 10.00 (s, 1H), 7.83 (s, 1H), 7.66 (s, 1H), 7.51 (s, 1H).

(7) Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (OTMS) CN

Znl ₂ (1.4 g; 4) was added to a solution of 3-chloro-5-trifluoromethoxybenzaldehyde (19.9 g; 88.6 mmol; see step (6) above) in CH ₂ Cl ₂ (600 ml) at 0 ° C. 4 mmol) and trimethylsilyl cyanide (9.7 g; 97.5 mmol). After stirring at 0 ° C for 1.5 hours and at room temperature for 2 hours, TLC analysis showed only the starting material. Znl _{2 was} added in portions until the reaction was complete (a total of more than 30.0 g Znl _{2 was} added). After stirring at room temperature for 18 hours, the reaction was quenched with water and the organic phase was separated. The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (27.7 g; 96%) as a liquid, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ): δ 7.43 (s, 1H), 7.28 (s, 1H), 7.25 (s, 1H), 5.49 (s, 1H), 0.38 (s, 9H).

(8) Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (OH) C (O) OH

Suspension Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (OTMS) CN (27.7 g; 85.6 mmol;

see step (7) above) in concentrated HCl (300 ml) was refluxed for 3 hours. The resulting brown heterogeneous mixture was cooled to room temperature and extracted with Et ₂ O (2x). The initial organic extracts were extracted with 2 N. NaOH (2x), then the main layer was acidified with 2 N. HCl and extracted with Et ₂ O. The Et ₂ O extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (4.9 g; 21%). TLC analysis of the initial organic extracts still showed the presence of the compound indicated in the subtitle, therefore, the main extraction / acidification procedure was repeated using 6 N NaOH to give an additional amount of the crude subtitled compound (2.8 g; 12%). TLC analysis of the initial organic extracts still showed the presence of the subtitled compound, so the organic extracts were dried (Na ₂ SO ₄ ) and concentrated in vacuo to give the sodium salt of the subtitled compound (18.3 g) as an oil. This salt was then redissolved in Et ₂ O and the organic phase acidified with 2 N HCl and washed with brine. The resulting organic phase was dried (Na ₂ SO ₄ ), treated with activated carbon, filtered through Celite® and concentrated in vacuo to give the crude subtitled compound (14.3 g; 62%) as a solid, which was used in the next step without further cleaning.

¹ H NMR (300 MHz; CD ₃ OD): δ 7.53 (s, 1H), 7.38 (s, 1H), 7.29 (s, 1H), 5.23 (s, 1H).

(9) Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) OH (a) and Ph (3-Cl) (5-OCF ₃ ) - (S) CH ( OAc) C (O) OH (b)

Mixture of Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (OH) C (O) OH (7.7 g; 28.5 mmol; see step (8) above) and Lipase PS " Amano "(3.8 g) in MTBE (100 ml) and vinyl acetate (50 ml) was stirred at 60 ° C for 26 hours. The reaction mixture was cooled, filtered through Celite® and the filter cake was washed with EtOAc. The combined organic phases were concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1) gave a mixture of the ammonium salts of the subtitle compound (a) and the subtitle compound (b) (6.7 g) and a pure sample of the ammonium salt of the subtitle compound (a) (1.2 g) with an enantiomeric excess (e.e.) of less than 95%. The appropriate fractions were dissolved in Et ₂ O and washed with 2 N. HCl (1x) and brine (1x), dried (Na ₂ SO ₄ ), filtered and concentrated to give the corresponding carboxylic acids (6.7 g and 1.1 g, respectively). Then, these fractions were individually re-converted to the conditions for separation and re-purified, as necessary, by chromatography on silica gel eluting with a mixture of CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1, or 75: 20: 5, or 145: 45: 10) as necessary. The purified subtitled compound (a) was acidified with aqueous HCl or aqueous citric acid before further use. The ammonium salt of the subtitle of compound (b) was used without characterization.

For the subtitle compound (a)

¹ H NMR (300 MHz; CD ₃ OD): δ 7.53 (s, 1H), 7.38 (s, 1H), 7.29 (s, 1H), 5.23 (s, 1H).

¹³ C NMR (75 MHz; CD ₃ OD): δ 174.9, 150.9, 145.4, 136.3, 126.8, 122.0, 120.6, 118.9, 72.9,

MS (m / z) 269 (M-1) ^- .

(10) Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

A solution of Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) OH (0.73 g 2.70 mmol; see step (9) above) in DMF (40 ml) in nitrogen atmosphere was cooled to 0 ° C. To this solution were added H-Aze-Pab (Teoc) (1.46 g; 3.24 mmol), collidine (0.82 g; 6.75 mmol) and RuBOP (1.83 g; 3.51 mmol). The solution was stirred at 0 ° C for 2 hours, warmed to room temperature and stirred for 18 hours, quenched with water and concentrated in vacuo. The residue was diluted with EtOAc and washed with H ₂ O (1x), aqueous NaHCO ₃ (1x), aqueous citric acid (1x) and brine (1x), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound. Flash chromatography on silica gel (2x), eluting with EtOAc / MeOH (30: 1), then CH ₂ Cl ₂ / MeOH (93: 7) gave the subtitled compound (0.73 g; 43%) as a crushable foam.

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers): δ 7.78-7.82 (d, 2H, J = 8 Hz), 7.25-7.54 (m, 5H), 5.25 and 5.16 (s, 1H), 5.22 and 4.79 (m, 1H), 3.92-4.58 (m, 6H), 2.20-2.76 (m, 2H), 1.04-1.13 (m, 2H), 0.08 (s, 9H).

MS (m / z) 629 (M + 1) ⁺ .

(11) Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab

Trifluoroacetic acid (1.0 ml) was added to a stirred solution cooled in an ice / water bath solution of Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) ( 101 mg; 160 μmol; see step (10) above) in methylene chloride (10 ml). The cooling bath was removed after 1 hour. After 1.5 hours at room temperature, acetonitrile (30 ml) was added and the solvents were carefully removed under reduced pressure. The residue was dissolved in water and freeze dried to give 90 mg (92%) of the title compound as its TFA salt.

MS (m / z) 483 (M-1) ^- ; 485 (M + 1) ⁺ .

¹ H NMR (300 MHz; CD ₃ OD) (complex spectrum due to the presence of diastereomers / rotamers): δ 7.70-7.80 (m, 2H), 7.45-7.58 (m, 3H), 7.24-7.38 (m, 2H), 5.26 (s, 1H), 5.17 (m, 1H, secondary rotamer), 4.82 (m, 1H, primary rotamer), 4.35-4.6 (m, 3H), 4.22 (m, 1H, primary rotamer), 3.92-4.12 (m , 2H, secondary rotamer), 2.70 (m, 1H, secondary rotamer), 2.55 (m, 1H, primary rotamer), 2.30 (m, 1H, primary rotamer), 2.16 (m, 1H, secondary rotamer).

¹³ C NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers): δ 173.7, 173.4, 173.0, 172.8, 168.1.

Example 7

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

HATU (71 mg; 0.19 mmol) was added to a stirred ice-water-cooled solution of Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) OH (39 mg; 0 , 14 mmol; see example 6 (9) above) in DMF (3 ml). After 30 minutes, a solution of H-Aze-Pab (OMe) × 2HCl (69 mg; 0.21 mmol; see international patent application WO 00/42059) and 2,4,6-collidine (0.080 ml; 0.58 mmol) were added. in DMF (1.5 ml). The reaction mixture was allowed to stand overnight, allowing the temperature to slowly rise to ambient temperature. The solvents were removed in vacuo and the crude product was purified using reverse phase HPLC (acetonitrile / 0.1 M aq. Ammonium acetate), which allowed to obtain, after drying, the corresponding fractions were frozen, the title compound (61 mg; 97%) as colorless solid.

MS (m / z) 513 (M-1) ^- ; 515 (M + 1) ⁺ .

¹ H NMR (500 MHz; CD ₃ OD) δ 7.97 (bt, 1H), 7.53 (d, 2H), 7.27 (t, 1H), 7.22 (d, 2H), 7.19 (t, 1H), 7.11 (t , 2H), 6.77 (s, 1H), 4.92 (s, 1H), 4.9 (bs, 3H), 4.81 (m, 2H), 4.40 (m, 2H), 4.09 (m, 1H), 3.87 (s, 3H), 2.58 (m, 1H), 2.37 (m, 1H).

¹³ C NMR (125 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 171.8, 169.9, 156.8.

Example 8

Parallel synthesis of alkoxyamidines

This synthesis was performed in a 96-well Robbins block. A solution of Ph (3-Cl) (5-OCF ₃ ) - (R) CH () was added to wells containing an appropriate amount of O-substituted hydroxylamine (specified below; all of the above are commercially available or obtained using well-known methods from the literature) OH) C (O) -Aze-Pab (Teos) (10 mg; 17 μmol; see Example 6 (10) above) in acetonitrile (1.0 ml). The block was sealed and the reaction mixture was rotated overnight in an oven at 60 ° C. After cooling and filtering, the solids were washed with acetonitrile (3 × 0.3 ml). The combined liquid fractions were concentrated in a vacuum centrifuge. The residue was partitioned between water (0.4 ml) and ethyl acetate (0.4 ml). At the end of the liquid-liquid extraction, everything was filtered through a Hydromatrix ™ column. After washing three times with ethyl acetate, the combined filtrates were concentrated in a vacuum centrifuge. Deprotection was carried out by adding methylene chloride (0.1 ml) and trifluoroacetic acid (0.3 ml). After stirring at room temperature for 3 hours, the solvents were removed in vacuo. The residue was partitioned between aqueous saturated sodium bicarbonate (0.5 ml) and ethyl acetate (0.5 ml). After extraction, filtration through Hydromatrix ™ and concentration (see below), the residue was dissolved in isopropanol / water (7: 3) (1 ml). Approximately 2% of this solution was taken and diluted with isopropanol / water (7: 3) (1 ml) for LC-MS analysis. After removing the solvents in vacuo, the solid residue was transferred to a 96-well plate using acetonitrile and ethyl acetate to dissolve the compound. The solvents were evaporated in a vacuum centrifuge, which allowed to obtain the following title compounds:

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH ₂ -3- (5-Me-isoxazole))

(from 3 - [(aminooxy) methyl] -5-methylisoxazole × HCl (18 mg; 0.11 mmol)), yield 3.64 mg (35%), MS (m / z) 596 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH2-3-pyridine)

(from 3 - [(aminooxy) methyl] pyridine × 2HCl (19 mg; 96 μmol), yield 5.14 mg (50%), MS (m / z) 592 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Bu)

(from O-isobutylhydroxylamine × HCl (17 mg; 140 μmol), yield 4.4 mg (45%), MS (m / z) 557 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OEt)

(from O-ethylhydroxylamine × HCl (14 mg; 140 μmol), yield 4.04 mg (42%), MS (m / z) 529 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn)

(from O-benzylhydroxylamine × HCl (17 mg; 110 μmol), yield 3.22 mg (29%), MS (m / z) 591 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclohexyl)

(from O-cyclohexylhydroxylamine × HCl (15 mg; 99 μmol), yield 2.9 mg (26%), MS (m / z) 583 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-cyclo-Bu)

(from O-cyclobutylhydroxylamine × HCl (17 mg; 140 μmol), yield 3.3 mg (30%), MS (m / z) 555 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OCH ₂ CH ₂ OPh (3-CF ₃ ))

(from O- [2- [3- (trifluoromethyl) phenoxy] ethyl] hydroxylamine × HCl (24 mg; 93 μmol), yield 6.52 mg (46%), MS (m / z) 689 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (4-Cl))

(from O- (4-chlorobenzyl) hydroxylamine × HCl (16 mg; 82 μmol), yield

3.47 mg (29%), MS (m / z) 625 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (3-MeO))

(from O- (3-methoxybenzyl) hydroxylamine × HCl (18 mg; 94 μmol), yield

4.33 mg (36%), MS (m / z) 621 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (2-Br))

(from O- (2-bromobenzyl) hydroxylamine × HCl (23 mg; 96 μmol), yield

3.87 mg (30%), MS (m / z) 671 (M + 1) ⁺ ;

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OBn (4-Me))

(from O- (4-methylbenzyl) hydroxylamine × HCl (14 mg; 81 μmol), yield

2.91 mg (25%), MS (m / z) 605 (M + 1) ⁺ and

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (O-4-heptyl)

(from O- (4-heptyl) hydroxylamine × HCl (15 mg; 89 μmol), yield 17 mg (100%), MS (m / z) 599 (M + 1) ⁺ .

Example 9

Ph (3-Cl) (5-OCHF ₂ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab × HOAc

(1) 3-Chloro-5-methoxybenzoic acid

Magnesium shavings (Fluka purum for Grignard reactions) were pretreated as follows. The chips were placed in a funnel of porous glass and 0.1 M hydrochloric acid was poured onto it. The chips were mixed with a glass rod for several seconds and then the acid was washed with 3 portions of water. Finally, the chips were washed with 2 portions of acetone and placed in a bottle. Tetrahydrofuran (100 ml; 99.95%) was dried by adding RedAI (1 g, 70 wt.% In toluene). Pre-treated magnesium chips (5 g, 200 mmol) were placed in a round bottom flask and purged with nitrogen 3 times. Dichloroanisole (26 g, 146 mmol) was dissolved in THF (100 ml dried using RedAI) and dibromethane (1.8 g; 10 mmol) was added. The reaction mixture was purged with nitrogen and then refluxed for 2 hours. The heating was interrupted and dry ice (10 g) was added portionwise over 2 minutes. When all the dry ice was dissolved, the reaction mixture was poured onto ice containing hydrochloric acid (400 ml, 2 M). Extractive treatment (diethyl ether, 300 ml) afforded 11.2 g (60.2 mmol; 41% yield) of the subtitle compound.

¹ H-NMR (500 MHz; acetone-d ₆ ): δ 7.57 (m, 1H), 7.49 (m, 1H), 7.23 (m, 1H), 3.91 (s, 3H).

(2) 3-Chloro-5-hydroxybenzoic acid

Alumina (1.65 g; 60 mmol) and iodine (21 g; 82 mmol) were refluxed in toluene (200 ml) for 2 hours. Then 3-chloro-5-methoxybenzoic acid (11.2 g; 60.2 mmol; see step (1) above) dissolved in toluene (50 ml) was added together with tetrabutylammonium iodide (1.5 g; 4 mmol), and this mixture was refluxed for another 2 hours. After cooling to ambient temperature, extractive treatment yielded 8.7 g (50 mmol; 83% yield) of the subtitle compound.

¹ H-NMR (300 MHz; acetone-d ₆ ): δ 9.27 (s, 1H), 7.48 (m, 1H), 7.44 (m, 1H), 7.11 (m, 1H).

(3) 3-Chloro-5-difluoromethoxybenzoic acid

3-Chloro-5-hydroxybenzoic acid (6.4 g; 37.2 mmol; see step (2) above) dissolved in chloroform (200 ml) was transferred to a 500 ml three-necked round-bottom flask equipped with an ice-cooled refrigerator and gas inlet pipe. Sodium hydroxide (100 ml, 5 M) was added with vigorous stirring. Chlorodifluoromethane (Freon 22; 25 g, 290 mmol) was added portionwise through a gas inlet tube at ambient temperature. After 2 hours, the reaction was completed. Extractive treatment yielded 6.2 g (28 mmol; 75% yield) of the subtitle compound.

¹ H-NMR (500 MHz; acetone-d ₆ ): δ 7.87 (m, 1H), 7.74 (m, 1H), 7.54 (m, 1H), 7.19 (t, 1H, J _HF = 73 Hz).

(4) 3-Chloro-5-difluoromethoxy-N-methoxy-N-methylbenzamide

3-Chloro-5-difluoromethoxybenzoic acid (1.8 g; 8 mmol; see step (3) above) and oxalyl chloride (1.5 g; 11.8 mmol) were dissolved in methylene chloride (50 ml). DMF (2 drops) was added and the reaction mixture was stirred at ambient temperature for 30 minutes. Then, N, O-dimethylhydroxylamine (1 g, 10.2 mmol) and triethylamine (3 g, 30 mmol) were added, and after stirring for another 10 minutes at ambient temperature, the reaction mixture was concentrated under reduced pressure. The residue was taken up in ether (100 ml) and water (50 ml). After separation, the organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated. This residue was chromatographed on silica (hexane / ethyl acetate (2: 1)) to give 2 g (7.5 mmol; 93%) of the subtitle compound.

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.54 (m, 1H), 7.37 (m, 1H), 7.27 (m, 1H), 6.53 (t, 1H, J _HF = 73 Hz).

(5) 3-Chloro-5-difluoromethoxyacetophenone

3-Chloro-5-difluoromethoxy-N-methoxy-N-methylbenzamide (2 g; 7.5 mmol; see step (4) above) was dissolved in ether (100 ml) and cooled in a stream of nitrogen to -70 ° C. Methyl lithium (7 ml, 11 mmol; 1.6 M in ether) was added dropwise to the stirred reaction mixture via syringe over 1 minute. The dry ice bath was removed and the mixture was left to reach ambient temperature before quenching the reaction with a solution of ammonium chloride (50 ml, 5% NH ₄ Cl in water). The organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexane / ethyl acetate (2: 1)) to give 1.5 g (6.8 mmol; 90% yield) of the subtitled compound.

¹ H-NMR (600 MHz; CDCl ₃ ): δ 7.77 (m, 1H), 7.59 (m, 1H), 7.35 (m, 1H), 6.56 (t, 1H, J _HF = 73 Hz), 2.60 (s , 3H).

(6) 3-Chloro-5-difluoromethoxyphenylacetic acid methyl ester

3-Chloro-5-difluoromethoxyacetophenone (1.5 g; 6.8 mmol; see step (5) above) was dissolved in methylene chloride (200 ml). Thallium (III) nitrate × 3MeOH on montmorillonite K-10 (6 g, 10 mmol (approximately 0.6 mmol / g)) was added; see J. Am. Chem. Soc., 98, 6750 (1976)) and the mixture was stirred at ambient temperature for 20 hours. The mixture was filtered and the filtrate was washed with sodium bicarbonate (100 ml; 0.5 M), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexane / ethyl acetate (2: 1)) to give 1 g (4 mmol; 56% yield) of the subtitled compound.

¹ H-NMR (500 MHz; CDCl ₃ ): δ 7.14 (m, 1H), 7.06 (m, 1H), 6.96 (m, 1H), 6.50 (t, 1H, J _HF = 73 Hz), 3.72 (s , 3H); 3.60 (s, 1H).

(7) α-Formyl (3-chloro-5-difluoromethoxyphenyl) acetic acid methyl ester

3-Chloro-5-difluoromethoxyphenylacetic acid methyl ester (1 g, 4 mmol; see step (6) above) and methyl formate (1 g, 16 mmol) were dissolved in ether (100 ml) and cooled in an ice bath (approximately 2 ° C) ) Finely chopped sodium (180 mg; 7.8 mmol) and methanol (1 ml) were then added and the mixture was allowed to stir in an ice bath overnight. Water (100 ml) was carefully added and the phases were separated. The water-containing phase was acidified with hydrochloric acid (2 M) to pH 1 and extracted with ether (2 × 100 ml). The extract was dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexane / ethyl acetate (1: 1)) to give 400 mg (1.4 mmol; 36% yield) of the subtitled compound.

¹ H-NMR (400 MHz): δ 12.10 (d, 1H), 7.32 (d, 1H), 7.11 (m, 1H), 7.07 (m, 1H), 6.94 (m, 1H), 6.51 (t, 1H , J _HF = 73 Hz), 3.83 (s, 3H).

(8) 3-Chloro-5-difluoromethoxytropic acid

α-Formyl (3-chloro-5-difluoromethoxyphenyl) acetic acid methyl ester (400 mg, 1.4 mmol; see step (7) above) was dissolved in a mixture of THF / methanol (50 ml, 9: 1). Sodium borohydride was added and the mixture was stirred at ambient temperature for 30 minutes. Water (100 ml) was added and the mixture was concentrated to form an aqueous suspension, which was converted into ethyl acetate and water. The phases were separated and the organic phase was washed with sodium chloride (15% in water), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in methanol (30 ml) and subjected to hydrolysis with sodium hydroxide (1 ml, 10 M) at ambient temperature for 10 minutes. Extractive work up gave 180 mg (0.68 mmol; 48% yield) of the subtitled compound.

¹ H-NMR (500 MHz; CDCl ₃ ): δ 7.18 (m, 1H), 7.10 (m, 1H), 7.00 (m, 1H), 6.50 (t, 1H, J _HF = 73 Hz), 4.11 (m , 1H), 3.90 (m, 1H), 3.84 (m, 1H).

(9) Ph (3-Cl) (5-OCHF ₂ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab × HOAc

3-Chloro-5-difluoromethoxytropic acid (180 mg; 0.7 mmol; see step (8) above), H-Aze-Pab (Teoc) × HCl (450 mg, 1 mmol) and RuBOP (530 mg, 1 mmol) ) was dissolved in DMF (10 ml), after which DIPEA (550 mg; 3.9 mmol) was added. The mixture was stirred at ambient temperature for 1 h before being diluted with brine (20 ml, 15% NaCl) and extracted with ethyl acetate (40 ml). The extract was dried over sodium sulfate, filtered and evaporated to dryness. The residue was dissolved in methylene chloride (5 ml) and trifluoroacetic acid (5 ml) was added. After 1 h at ambient temperature, the mixture of diastereomers was evaporated to dryness and the residue was chromatographed on a reverse phase column (acetonitrile / water (30:70), buffer: 0.1 M ammonium acetate). Freeze-drying gave 36 mg (0.067 mmol; 10.4% yield) of the title compound.

MS (ES) 481 (M + 1) ⁺ .

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.77 (d, 2H), 7.57 (d, 2H), 7.30 (m, 1H), 7.13 (m, 2H), 6.87 (t, 1H, J _HF = 73 Hz), 4.76 (m, 1H), 4.55 (s, 2H), 4.37 (m, 1H), 4.03 (m, 2H), 3.82 (m, 1H), 3.72 (m, 1H), 2.53 (m, 1H), 2.28 (m, 1H), 1.92 (3, 1.5H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 172.3, 171.9, 167.2.

Example 10

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab × TFA

(1) 3-Chloro-5-trifluoromethoxybenzyl mesylate

To a solution of 3-chloro-5-trifluoromethoxybenzyl alcohol (6.1 g; 26.9 mmol; see example 6 (5) above) in CH ₂ Cl ₂ (250 ml) at 0 ° C under nitrogen atmosphere was added DIPEA (4, 2 g; 32.3 mmol) and methanesulfonyl chloride (3.4 g; 29.6 mmol). The solution was stirred at 0 ° C for 1.5 hours and quenched with H ₂ O. The organic layers were separated and washed with H ₂ O (1x), 1 N. HCl (1x), H ₂ O (1x) and aqueous NaHCO ₃ (1x) and then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (8.2 g; 99%) as oils.

¹ H NMR (300 MHz; CDCl ₃ ): δ 7.37 (s, 1H), 7.28 (s, 1H), 7.18 (s, 1H), 5.23 (s, 2H), 3.07 (s, 3H).

(2) 3-Chloro-5-trifluoromethoxybenzyl cyanide

To a solution of 3-chloro-5-trifluoromethoxybenzylmesylate (8.2 g; 26.8 mmol; see step (1) above) in DMSO (50 ml) was added sodium cyanide (2.6 g; 53.6 mmol). The resulting heterogeneous solution was heated to 50 ° C and sonicated for 1 hour. The reaction mixture was cooled and partitioned between Et ₂ O and H ₂ O. The organic phase was washed with H ₂ O (2x) and brine (2x). The combined aqueous phases were extracted with Et ₂ O (1x). The combined organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated under mild heating and partial vacuum, which afforded the subtitle compound (6.3 g; 100%) as a reddish volatile oil, which was used in the next step without further cleaning up.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.32 (s, 1H), 7.24 (s, 1H), 7.12 (s, 1H), 3.78 (s, 2H).

(3) 3-Chloro-5-trifluoromethoxyphenylacetic acid

To a solution of 3-chloro-5-trifluoromethoxybenzyl cyanide (6.3 g; 26.7 mmol; see step (2) above) in 2-propanol (100 ml) was added water (200 ml) and sodium hydroxide (7.5 g; 133.5 mmol). The solution was refluxed for 18 hours, cooled to room temperature, and 2-propanol was removed in vacuo. The aqueous phase was washed with CH ₂ Cl ₂ (2x) and the washings were discarded. The main aqueous phase was acidified with 2 N. HCl and was extracted with CH ₂ Cl ₂ (3x). The CH ₂ Cl ₂ extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (5.2 g; 76%) as an oil, which was used in the next step without further purification.

¹ H NMR (300 MHz; CDCl ₃ ): δ 7.25 (s, 1H), 7.19 (s, 1H), 7.08 (s, 1H), 3.68 (s, 2H).

(4) Ethyl 3-chloro-5-trifluoromethoxyphenyl acetate

To a solution of 3-chloro-5-trifluoromethoxyphenylacetic acid (5.2 g; 20.4 mmol; see step (3) above) in EtOH (600 ml) was added sulfuric acid (a few drops). The solution was refluxed for 18 h, cooled to room temperature, neutralized with solid NaHCO ₃ and EtOH was removed in vacuo. The residue was diluted with EtOAc, then washed with H ₂ O (1x), aqueous NaHCO ₃ (1x) and brine (1x). The organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (5.5 g; 96%) as an oil, which was used in the next step without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.24 (s, 1H), 7.16 (s, 1H), 7.07 (s, 1H), 4.13-4.22 (q, J = 8 Hz, 2H), 3.63 (s, 2H), 1.24-1.32 (t, J = 8 Hz, 3H).

(5) Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (CHO) C (O) OEt

To a solution of ethyl 3-chloro-5-trifluoromethoxyphenyl acetate (4.5 g; 15.9 mmol; see step (4) above) in anhydrous THF (400 ml) in a nitrogen atmosphere at a temperature of less than 0 ° C (in an ice bath MeOH) sodium ethoxide (4.5 g; 63.6 mmol) was added. The cooled solution was stirred for 40 minutes and ethyl formate (8.1 g; 111.3 mmol) was added. The solution was stirred at 0 ° C for 30 minutes, warmed to room temperature and stirred for 2 hours. Then THF was removed in vacuo. The residue was diluted with Et ₂ O and extracted with H ₂ O (1x) and 0.5 M NaOH (3x). The aqueous extracts were acidified with 2 N. HCl and was extracted with CH ₂ Cl ₂ (3x). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound (3.9 g). Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (4: 1) afforded the subtitled compound (3.0 g; 61%) as an oil.

¹ H NMR (300 MHz; CDCl ₃ ; mixture of isomers): δ 12.30 and 12.25 (s, 1H), 7.39 and 7.34 (s, 1H), 7.21 (s, 1H), 7.17 (s, 1H), 7.08 (s , 1H), 4.27-4.37 (q, J = 8 Hz, 2H), 1.28-1.38 (t, J = 8 Hz, 3H).

(6) Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (CH ₂ OH) C (O) OEt

To a solution of Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (CHO) C (O) OEt (3.0 g; 9.66 mmol; see step (5) above) in MeOH ( 200 ml) at -10 ° C (in an ice-MeOH bath) sodium borohydride (0.7 g; 19.32 mmol) was added in portions over 5 min. The solution was stirred at −10 ° C. for 45 minutes and an additional amount of sodium borohydride (0.4 g) was added. After another 15 minutes, the reaction was quenched with aqueous ammonium chloride, the mixture was made slightly acidic using 2 N. HCl and MeOH were removed in vacuo. The residue was diluted with EtOAc and washed with H ₂ O (1x), aqueous NaHCO ₃ (1x) and brine (1x). The organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the crude subtitled compound. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (5: 1) afforded the subtitled compound (2.0 g; 66%) as an oil.

¹ H NMR (300 MHz; CDCl ₃ ): δ 7.26 (s, 1H), 7.19 (s, 1H), 7.07 (s, 1H), 4.16-4.28 (m, 2H), 4.04-4.15 (m, 1H) , 3.76-3.94 (m, 2H), 2.33 (t, J = 6 Hz, 1H), 1.18-1.30 (t, J = 8 Hz, 3H).

(7) Ph (3-Cl) -OCF ₃ ) - (R, S) CH (CH ₂ OH) C (O) OH

To a solution of Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (CH ₂ OH) C (O) OEt (2.0 g; 6.24 mmol; see step (6) above) in THF (50 ml) and H ₂ O (25 ml) was added lithium hydroxide monohydrate (0.5 g; 12.48 mmol). The solution was stirred at room temperature for 1 hour and THF was removed in vacuo. The residue was diluted with H ₂ O, then washed with CHCl ₃ (2x) and the washings were discarded. The main aqueous layer was acidified with 2 N. HCl and extracted with CHCl ₃ (4x). The CHCl ₃ extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the crude subtitled compound (1.5 g) as an oil. Flash chromatography on silica gel with elution with a mixture of CHCl ₃ / MeOH / concentrated NH ₄ OH (gradient from 7.0: 2.5: 0.5 to 6: 3: 1) yielded the ammonium salt of the subtitle compound (1.1 d). Ammonium salt was distributed between 1 N. HCl and CHCl ₃ . The organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (also known as 3-chloro-5-trifluoromethoxytropic acid) as an oil (1.1 g; 62%).

¹ H NMR (300 MHz; CD ₃ OD): δ 7.41 (s, 1H), 7.27 (s, 1H), 7.24 (s, 1H), 4.03 (m, 1H), 3.75-3.87 (m, 2H).

(8) Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (Teoc) (a) and Ph (3-Cl) (5-OCF ₃ ) - (R) CH (CH ₂ OH) C (O) -Aze-Pab (Teoc) (b)

To a solution of Ph (3-Cl) (5-OCF ₃ ) - (R, S) CH (CH ₂ OH) C (O) OH (0.65 g; 2.28 mmol; see step (7) above) in DMF (40 ml) at a temperature below 0 ° C (in an ice-MeOH bath) was added H-Aze-Pab (Teoc) (0.90 g; 2.39 mmol), collidine (0.71 g; 5.70 mmol) ) and RuVOR (1.31 g; 2.51 mmol). The resulting solution was stirred at a temperature below 0 ° C for 1 h, warmed to room temperature and stirred for 1 h. DMF was then removed in vacuo. The residue was diluted with EtOAc and washed with dilute aqueous HCl (1x), brine (1x), aqueous NaHCO ₃ (1x) and brine (1x). The organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the crude subtitled compound (2.1 g) as a mixture of diastereomers. Flash chromatography (3x) on silica gel, eluting first with EtOAc / MeOH (95: 5), then CH ₂ Cl ₂ / MeOH (97: 3) and finally CH ₂ Cl ₂ / MeOH (95: 5), allowed to obtain the subtitle of the compound is diastereomer (a) (0.51 g; 35%) and diastereomer (b) (0.45 g; 31%) as a crushable foam.

For the diastereomer compound (a) indicated in the subtitle

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.79-7.85 (d, J = 8 Hz, 2H), 7.22-7.49 (m, 5H), 5.17-4.77 (m, 1H), 4.53-4.18 (m, 4H), 3.58-4.11 (m, 5H), 2.47-2.73 (m, 1H), 2.11-2.34 (m, 1H), 1.08-1.12 (m, 2H), 0.07 (s, 9H )

MS (m / z) 643 (M + 1) ⁺ .

(9) Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (Teoc) (78 mg; 0.121 mmol; see step (8) above - diastereomer (a )) was dissolved in 5 ml of trifluoroacetic acid. After 10 minutes, the reaction was completed and the solvent was evaporated. The residue was dried by freezing from water and acetonitrile to obtain the desired product. Yield 70 mg (94%).

MS (m / z) 483 (M-1) ^- ; 485 (M + 1) ⁺

¹ H-NMR (400 MHz, D ₂ O) rotamers (1: 1): δ 8.83 (bt, 1H), 7.79 (d, 1H), 7.72 (d, 1H), 7.54 (d, 1H), 7.43 ( d, 2H), 7.35 (m, 1H, rotamer), 7.28 (m, 1H, rotamer), 7.20 (m, 1H, rotamer), 7.05 (m, 1H, rotamer), 5.22 (m, 1H, rotamer), 4.83 (m, 1H, rotamer), 4.57 (m, 2H, rotamer), 4.38 (m, 2H, rotamer), 4.3-3.7 (m, 5H), 2.77 (m, 1H, rotamer), 2.55 (m, 1H , rotamer), 2.27 (m, 1H).

¹³ C-NMR (100 MHz; D ₂ O) (carbonyl and / or amidine carbons, rotamers) δ 172.9, 172.2, 172.0, 171.8, 166.9.

Example 11

Ph (3-Cl) (5-OCF ₃ ) - (3) CH (CH ₂ OH) C (O) -Aze-Pab (OMe)

(1) Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (Ohm, Teoc)

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (Teos) (100 mg; 0.155 mmol; see example 10 (8) above) was dissolved in 12 ml of tetrahydrofuran. O-methylhydroxylamine hydrochloride (44 mg; 0.53 mmol) was added and the reaction mixture was heated at 50 ° C. overnight. The reaction mixture was evaporated and the residue was purified by preparative HPLC (CH ₃ CN / 0.1 M NH ₄ OAC (70:30)). The appropriate fractions were evaporated, the residue was dissolved in a small amount of acetonitrile and water and dried by freezing. Freeze-drying was repeated again. Yield 80 mg (76%) of pure material.

¹ H-NMR (400 MHz; CD ₃ OD; rotamers): δ 7.5-7.4 (m, 3H), 7.35-7.2 (m, 4H), 5.15 (m, 1H, minor rotamer), 4.74 (m, 1H, main rotamer), 4.5-4.25 (m, 3H), 4.2-3.95 (m, 4H), 3.91 (b, 3H), 3.9-3.6 (m, 2H), 2.63 (m, 1H, minor rotamer), 2.50 ( m, 1H, main rotamer), 2.3-2.1 (m, 1H), 0.95 (m, 2H), 0.02 (s, 9H, main rotamer), 0.01 (s, 9H, minor rotamer).

(2) Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (OMe)

Ph (3-Cl) (5-OCF ₃ ) - (S) CH (CH ₂ OH) C (O) -Aze-Pab (OMe, Teoc) (80 mg; 0.12 mmol; see step (1) above) ) was dissolved in 1 ml of methylene chloride and cooled in an ice bath. 3 ml of trifluoroacetic acid was added and the reaction vessel was kept in an ice bath for two hours. The mixture was evaporated, dissolved in ethyl acetate and washed three times with NaHCO ₃ (aq), then with water and brine. The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated. The residue was freeze-dried from a small amount of acetonitrile and water. Yield 60 mg (95%) of the pure title product.

MS (m / z) 528 (M-1) ^- ; 531 (M + 1) ⁺ .

¹ H-NMR (500 MHz; CD ₃ OD; rotamers): δ 7.65-7.55 (m, 3H, rotamers), 7.45 (m, 1H, main rotamer), 7.4-7.2 (m, 4H), 5.15 (m, 1H, minor rotamer), 4.74 (m, 1H, main rotamer), 4.5-4.3 (m, 3H), 4.05-3.95 (m, 2H), 3.85 (m, 1H, main rotamer), 3.82 (s, 3H, main rotamer), 3.81 (s, 3H, minor rotamer), 3.73 (m, 1H, minor rotamer), 3.67 (m, 1H, minor rotamer), 3.62 (m, 1H, minor rotamer), 2.63 (m, 1H, minor rotamer), 2.50 (m, 1H, main rotamer), 2.24 (m, 1H, main rotamer), 2.16 (m, 1H, minor rotamer).

¹³ C-NMR (125 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 174.0, 173.2, 172.7, 172.6, 155.1.

Example 12

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe. Teoc)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.40 g; 0.65 mmol; see example 1 (9) above) dissolved in 20 ml of acetonitrile and 0.50 g (6.0 mmol) of O-methylhydroxylamine hydrochloride was added. The mixture was heated at 70 ° C. for 2 hours. The solvent was evaporated and the residue was partitioned between water and ethyl acetate. The aqueous phase was extracted twice more with ethyl acetate, the combined organic phase was washed with water, brine, dried (Na ₂ SO ₄ ), filtered and evaporated. Yield 0.41 g (91%).

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.83 (bt, 1H), 7.57 (bs, 1H), 7.47 (d, 2H), 7.30 (d, 2H), 7.20 (m, 1H), 7.14 ( m, 1H), 7.01 (m, 1H), 6.53 (t, 1H), 4.89 (s, 1H), 4.87 (m, 1H), 4.47 (m, 2H), 4.4-4.2 (b, 1H), 4.17 -4.1 (m, 3H), 3.95 (s, 3H), 3.67 (m, 1H), 2.68 (m, 1H), 2.42 (m, 1H), 0.97 (m, 2H), 0.01 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe, Teoc) (0.40 g; 0.62 mmol; see step (1) above) ) was dissolved in 5 ml of TFA and allowed to interact for 30 minutes. TFA was evaporated and the residue was partitioned between ethyl acetate and NaHCO ₃ (aq). The aqueous phase was extracted twice more with ethyl acetate, the combined organic phase was washed with water, brine, dried (Na ₂ SO ₄ ), filtered and evaporated. The product was dried by freezing from a mixture of water / acetonitrile. No cleaning was required. Yield 0.28 g (85%).

¹ H-NMR (600 MHz; CDCl ₃ ): δ 7.89 (bt, 1H), 7.57 (d, 2H), 7.28 (d, 2H), 7.18 (m, 1H), 7.13 (m, 1H), 6.99 ( m, 1H), 6.51 (t, 1H), 4.88 (s, 1H), 4.87 (m, 1H), 4.80 (bs, 2H), 4.48 (dd, 1H), 4.43 (dd, 1H), 4.10 (m , 1H), 3.89 (s, 3H), 3.68 (m, 1H), 2.68 (m, 1H), 2.40 (m, 1H).

¹³ C-NMR (125 MHz; CDCl ₃ ) (carbonyl and / or amidine carbons, rotamers) δ 172.9, 170.8, 152.7, 152.6.

MS (m / z) 495 (M-1) ^- , 497 (M + 1) ⁺ .

Example 13

Ph (3-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × HOAc

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe) (13 mg; 0.026 mmol; see example 12 above) was dissolved in absolute ethanol (5 ml ) and 30 mg of 10% Pd / C was added. At the end, acetic acid (5 μl) was added and the mixture was hydrogenated at atmospheric pressure for 20 hours. The mixture was filtered through Celite®, evaporated and purified by reverse phase HPLC (0.1 M aqueous ammonium acetate / MeCN). The appropriate fractions were freeze dried, which afforded the title compound as a white solid (8.5 mg, 66%).

¹ H-NMR (400 MHz; CD ₃ OD, rotamers): δ 7.73-7.78 (m, 2H), 7.55 (d, 2H), 7.19-7.43 (m, 3H), 7.06-7.13 (m, 1H), 6.83 (t, 1H, J _HF = 74 Hz, main rotamer), 6.81 (t, 1H, main rotamer), 5.20 (s, 1H, main rotamer), 5.19 (m, 1H, minor rotamer), 5.15 (s, 1H, minor rotamer), 4.78 (m, 1H, main rotamer), 4.4-4.6 (several peaks, 2H), 4.35 (m, 1H, main rotamer), 4.08 (m, 1H), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.52 (m, 1H, main rotamer), 2.30 (m, 1H, main rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 173.7, 172.9, 168.3.

MS (m / z) 433 (M + 1) ⁺ ; 431 (M-1) ^- .

Example 14

Ph (3-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCF ₃ ) - (R) CH (OH) C (O) -Aze-Pab × TFA (34 mg; 0.057 mmol; from Example 6) was dissolved in 5 ml of ethanol and 20 mg was added 10% Pd / C. The mixture was hydrogenated at atmospheric pressure overnight. The mixture was filtered through Celite®, evaporated and dried by freezing from a mixture of water / acetonitrile.

¹ H-NMR (400 MHz; CD ₃ OD; rotamers): δ 7.8-7.7 (m, 2H), 7.55 (m, 2H), 7.5-7.2 (m, 4H), 5.24 (s, 1H, main rotamer) , 5.23 (m, 1H, minor rotamer), 5.18 (s, 1H, minor rotamer), 4.77 (m, 1H, main rotamer), 4.6-4.45 (m, 2H), 4.36 (m, 1H, main rotamer), 4.08 (m, 1H), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.52 (m, 1H, main rotamer), 2.30 (m, 1H, main rotamer), 2.15 (m , 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 174.1, 173.9, 173.5, 172.9, 168.2.

¹⁹ F-NMR (282 MHz; CD ₃ OD): -59.8 and -59.9 (3F, minor and major rotamers, respectively), -77.4 (3F) indicates that it is a TFA salt.

MS (m / z) 451.3 (M + 1) ⁺ .

Example 15

Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 3-Chloro-5-trifluoroethoxybenzaldehyde

To a stirred solution of 3-chloro-5-hydroxybenzaldehyde (2.0 g; 12.8 mmol; see example 1 (2) above) and potassium carbonate (2.3 g; 16.6 mmol) in DMF (35 ml) 2,2,2-trifluoroethyl-p-toluenesulfonate (4.2 g; 16.6 mmol) was added under nitrogen atmosphere at room temperature. The mixture was heated to 110 ° C. for 7 hours and then stirred overnight at room temperature. The reaction mixture was cooled to 0 ° C, poured into chilled in ice 2 N. HCl (100 ml) and was extracted with EtOAc (2 × 75 ml). The combined organic extracts were washed with 0.5 N. HCl (2 × 50 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with a Hex / EtOAc mixture (6: 1), which afforded the subtitled compound (1.9 g; 61%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.44 (s, 1H), 7.56 (s, 1H), 7.33 (s, 1H), 7.28 (s, 1H), 4.42 (q, J = 8 Hz, 2H) .

(2) Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R, S) CH (OTMS) CN

To a solution of 3-chloro-5-trifluoroethoxybenzaldehyde (5.2 g; 21.7 mmol; see step (1) above) and zinc iodide (1.7 g; 5.4 mmol) in CH ₂ Cl ₂ (200 ml) in a nitrogen atmosphere, trimethylsilyl cyanide (4.3 g; 43.3 mmol) was added dropwise with a syringe at 0 ° C. The mixture was stirred at 0 ° C. for 3 hours, then diluted with H ₂ O (150 ml). The organic layer was separated, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (6.9 g; 95%) as a yellow oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.27 (s, 1H), 6.98 (s, 2H), 5.44 (s, 1H), 4.38 (q, J = 8 Hz, 2H), 0.30 (s, 9H) .

(3) Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R, S) CH (OH) C (O) OH

Concentrated hydrochloric acid (170 ml) was added to Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R, S) CH (OTMS) CN (6.9 g; 20.4 mmol; see step (2) above) and stirred at 100 ° C for 1 h. After cooling to room temperature, the reaction mixture was further cooled to 0 ° C and slowly made alkaline with 3 N NaOH (300 ml). This mixture was washed with Et ₂ O (2 × 100 ml) and the aqueous layer was acidified with 2 N. HCl (50 ml). The aqueous layer was then extracted with EtOAc (2 × 100 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (5.3 g; 92%) as a pale yellow oil, which used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.18 (s, 1H), 7.07 (s, 1H), 7.02 (s, 1H), 5.13 (s, 1H), 4.58 (q, J = 8 Hz, 2H )

(4) Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) OH (a) and Ph (3-Cl) (5-OCH ₂ CF ₃ ) - ( S) CH (OAc) C (O) OH (b)

Solution Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R, S) CH (OH) C (O) OH (7.06 g; 24.8 mmol; see step (3) above) and Lipases PS "Amano" (4.30 g) in vinyl acetate (250 ml) and MTBE (250 ml) was heated at 70 ° C. under nitrogen for 40 hours. The reaction mixture was cooled to room temperature, the enzyme was removed by filtration with EtOAc washing and the filtrate was concentrated in vacuo. Silica gel chromatography, eluting with CHCl ₃ / MeOH / Et ₃ N (92: 6: 2), afforded the subtitled triethylamine salt of the subtitled compound (a) (3.02 g) as a yellow oil. The salt of the subtitle compound (a) was dissolved in H ₂ O (150 ml), acidified with 2 N. HCl and extracted with EtOAc (2 × 75 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (a) (2.18 g) as an off-white solid. In addition, after carrying out the above column chromatography, a triethylamine salt of the subtitle compound (b) (4.73 g) was obtained.

Data for the subtitled compound (a)

Mp 98-103 ° C.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.18 (s, 1H), 7.07 (s, 1H), 7.02 (s, 1H), 5.13 (s, 1H), 4.58 (q, J = 8 Hz, 2H )

¹³ C NMR (75 MHz; CD ₃ OD) δ 175.4, 159.6, 144.6, 136.2, 125.0 (q, J = 277 Hz), 121.8, 115.9, 113.1, 73.3, 67.0 (q, J = 35 Hz).

HPLC analysis: 98.6%,> 99% e.e., Chiralcel OD column (Hex / EtOH / TFA mobile phase (97: 3: 0.5)).

[α] ²⁵ _D = -81.5 ° (c = 1.0, MeOH).

APCl-MS: (M-1) = 283 m / z.

(5) Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) OH (0.50 g; 1.8 mmol; see step (4) above (compound (a) ))) in DMF (20 ml) under nitrogen atmosphere was added H-Aze-Pab (Teoc) × HCl (1.03 g; 2.3 mmol), RuBOP (1.01 g; 1.9 mmol) and DIPEA ( 0.57 g; 4.4 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo, the residue was chromatographed twice on silica gel, eluting first with CHCl ₃ / EtOH (10: 1) and then EtOAc / EtOH ( 10: 1), which allowed to obtain the compound indicated in the subtitle (0.55 g; 48%) in the form of crushed white foam.

Mp 90-95 ° C.

R _f = 0.42 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.78-7.81 (m, 2H), 7.38-7.41 (m, 2H), 7.12-7.16 (m, 1H), 7.00-7.06 (m, 2H ), 5.09-5.22 and 4.75-4.79 (m, 2H), 3.94-4.61 (m, 8H), 2.09-2.75 (m, 2H), 1.04-1.11 (m, 2H), 0.70 (s, 9H).

APCl-MS: (M + 1) = 643 m / z.

(6) Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.066 g; 0.103 mmol; see step (5) above) was dissolved in 3 ml of TFA and allowed to interact for 30 minutes. TFA was evaporated and the residue was freeze-dried from water / acetonitrile to give 0.060 g (94%) of the title compound as its TFA salt.

¹ H-NMR (400 MHz; CD ₃ OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.2-7.0 (m, 3H), 5.21 (m, 1H, minor rotamer ), 5.17 (s, 1H, main rotamer), 5.11 (s, 1H, minor rotamer), 4.81 (m, 1H, main rotamer), 4.6-4.4 (m, 4H), 4.37 (m, 1H, main rotamer) , 4.16 (m, 1H, main rotamer), 4.06 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.54 (m, 1H, main rotamer) 2.29 (m, 1H, main rotamer); 2.15 (m, 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.2, 171.8, 171.7, 167.0.

MS (m / z) 499.3 (M + 1) ⁺ .

Example 16

Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

To a solution of Ph (3-Cl) (5-OCH ₂ CF ₃ ) - (R) CH (OH) C (O) OH (0.48 g; 1.7 mmol; see example 15 (4) above (compound ( a))) in DMF (20 ml) under nitrogen atmosphere was added H-Aze-Pab (OMe) × 2HCl (0.74 g; 2.2 mmol), PyBOP (0.97 g; 1.9 mmol) and DIPEA (0.55 g; 4.2 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo, the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (10: 1) and a second time with EtOAc / EtOH (10: 1), which afforded the title compound (0.62 g; 69%) as a crushable white foam.

Mp 75-80 ° C.

R _f = 0.43 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.57-7.60 (m, 2H), 7.32-7.36 (m, 2H), 7.13-7.17 (m, 1H), 7.00-7.06 (m, 2H ), 5.09-5.19 and 4.74-4.80 (m, 2H), 3.93-4.62 (m, 6H), 3.81 (s, 3H), 2.10-2.73 (m, 2H).

APCl-MS: (M + 1) = 529 m / z.

Example 17

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) methanesulfonic acid 2,2-difluoroethyl ester

To a stirred magnetic stirrer solution of 2,2-difluoroethanol (1.52 g; 18.5 mmol) in CH ₂ Cl ₂ (20 ml) in a nitrogen atmosphere were added triethylamine (5.61 g; 55.5 mmol) and methanesulfonyl chloride ( 2.54 g; 22.2 mmol) at 0 ° C. The mixture was stirred at 0 ° C for 1.5 h, diluted with CH ₂ Cl ₂ (50 ml) and washed with 2 N. HCl (50 ml). The aqueous layer was extracted with CH ₂ Cl ₂ (30 ml), the combined organic extracts were washed with brine (30 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (2.52 g; 85%) as a yellow oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 6.02 (tt, J = 3 Hz, J = 55 Hz, 1H), 4.39 (dt, J = 3 Hz, J = 13 Hz, 2H), 3.13 (s, 3H )

(2) 3-Chloro-5-difluoroethoxybenzaldehyde

To a solution of 3-chloro-5-hydroxybenzaldehyde (1.50 g; 9.6 mmol; see example 1 (2) above) and potassium carbonate (1.72 g; 12.5 mmol) in DMF (10 ml) in the atmosphere nitrogen was added dropwise a solution of methanesulfonic acid 2,2-difluoroethyl ester (2.0 g; 12.5 mmol; see step (1) above) in DMF (10 ml) at room temperature. The mixture was heated to 100 ° C. for 6 hours, then stirred overnight at room temperature. The reaction mixture was cooled to 0 ° C, poured into chilled in ice 2 N. HCl (100 ml) and was extracted with EtOAc (2 × 75 ml). The combined organic extracts were washed with 0.5 N. HCl (2 × 50 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with a Hex / EtOAc mixture (5: 1), which afforded the subtitled compound (1.35 g; 64%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.92 (s, 1H), 7.52 (s, 1H), 7.31 (s, 1H), 7.22 (s, 1 H), 6.12 (tt, J = 3 Hz, J = 55 Hz, 1H), 4.26 (dt, J = 3 Hz, J = 15 Hz, 2H).

(3) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R, S) CH (OTMS) CN

To a solution of 3-chloro-5-difluoroethoxybenzaldehyde (1.35 g; 6.1 mmol; see step (2) above) and zinc iodide (0.48 g; 1.5 mmol) in CH ₂ Cl ₂ (50 ml) under nitrogen atmosphere, trimethylsilyl cyanide (1.21 g; 12.2 mmol) was added dropwise. The mixture was stirred at 0 ° C for 3 hours, then diluted with H ₂ O (50 ml). The organic layer was separated, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (1.85 g; 95%) as a brown oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.13 (s, 1H), 6.94 (s, 2H), 6.10 (tt, J = 3 Hz, J = 55 Hz, 1H), 5.43 (s, 1H), 4.20 (dt, J = 3 Hz, J = 15 Hz, 2H), 0.28 (s. 9H).

(4) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R, S) CH (OH) C (O) OH

Concentrated hydrochloric acid (60 ml) was added to Ph (3-Cl) (5-OCH ₂ CHF ₃ ) - (R, S) CH (OTMS) CN (1.85 g; 5.8 mmol; see step (3) above) and stirred at 100 ° C for 1 h. After cooling to room temperature, the reaction mixture was further cooled to 0 ° C, slowly made alkaline with 3 N NaOH (~ 180 ml) and washed with Et ₂ O (2 × 75 ml). The aqueous layer was acidified with 2 N. HCl (20 ml) and was extracted with EtOAc (2 × 75 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to afford the subtitled compound (1.50 g; 97%) as a pale yellow oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.15 (s, 1H), 7.05 (s, 1H), 6.98 (s, 1H), 6.19 (tt, J = 4 Hz, J = 55 Hz, 1 N) 5.12 (s, 1 H), 4.25 (dt, J = 4 Hz, J = 17 Hz, 2H).

(5) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (S) CH (OAc) C (O) OH (a) and Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - ( R) CH (OH) C (O) OH (b)

Solution Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R, S) CH (OH) C (O) OH (3.90 g; 14.6 mmol; see step (4) above) and Lipases PS "Amano" (2.50 g) in vinyl acetate (140 ml) and MTBE (140 ml) was heated at 70 ° C. under nitrogen for 40 hours. The reaction mixture was cooled to room temperature, the enzyme was removed by filtration with washing with EtOAc and the filtrate was concentrated in vacuo. Silica gel chromatography, eluting with CHCl ₃ / MeOH / Et ₂ N (92: 6: 2), afforded the triethylamine salt of the subtitle compound (a) as a yellow oil. In addition, a triethylamine salt of the subtitle compound (b) (1.47 g) was obtained, this salt was dissolved in H ₂ O (100 ml), acidified with 2 N HCl and extracted with EtOAc (2 × 75 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (b) (1.00 g) as an off-white solid.

Data for the subtitle compound (b)

Mp 103-106 ° C.

R _f = 0.39 (CHCl ₃ / MeOH / Et ₂ N (90: 8: 2)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.13 (s, 1H), 7.04 (s, 1H), 6.97 (s, 1H), 6.17 (tt, J = 4 Hz, J = 55 Hz, 1H), 5.12 (s, 1 H), 4.24 (dt, J = 4 Hz, J = 8 Hz, 2H).

¹³ C NMR (75 MHz; CD ₃ OD) δ 175.5, 160.3, 144.5, 136.1, 121.3, 115.7, 115.3 (t, J = 240 Hz), 112.9, 73.4, 68.6 (t, J = 29 Hz).

HPLC analysis: 96.2%,> 95.0% e.e., ChiralPak AD column (Hex / EtOH / TFA mobile phase (95: 5: 0.5)).

[α] ²⁵ _D = -84.0 ° (c = 0.85, MeOH).

APCl-MS: (M-1) = 265 m / z.

(6) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) OH (0.35 g; 1.3 mmol; see step (5) above (compound (b) ))) in DMF (18 ml) under nitrogen atmosphere, H-Aze-Pab (Teoc) × HCl (0.76 g; 1.7 mmol), PyBOP (0.75 g; 1.4 mmol) and DIPEA ( 0.43 g; 3.3 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo and the residue was chromatographed twice on silica gel, eluting first with CHCl ₃ / EtOH (10: 1) and then EtOAc / EtOH ( 10: 1), which made it possible to obtain the compound indicated in the subtitle (0.69 g; 84%) as a crushable white foam.

Mp 108-118 ° C.

R _f = 0.48 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.78-7.81 (m, 2H), 7.40-7.43 (m, 2H), 7.09-7.12 (m, 1H), 6.96-7.02 (m, 2H ), 6.16 (t, J = 57 Hz, 1H), 5.09-5.20 and 4.75-4.80 (m, 2H), 3.95-4.55 (m, 8H), 2.10-2.75 (m, 2H), 1.04-1.11 (m, 2H), 0.07 (s, 9H).

APCl-MS: (M + 1) = 625 m / z.

(7) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.086 g; 0.138 mmol; see step (6) above) was dissolved in 3 ml of TFA and allowed to react for 1 h. TFA was evaporated and the residue was dried by freezing from a mixture of water / acetonitrile to obtain 0.080 g (98%) of the title compound as its TFA salt.

¹ H-NMR (300 MHz; CD ₃ OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.15-6.95 (m, 3H), 6.35-5.95 (m, 1H) 5.20 (m, 1H, minor rotamer), 5.14 (s, 1H, primary rotamer), 5.10 (s, 1H, minor rotamer), 4.80 (m, 1H, primary rotamer), 4.6-4.0 (m, 6H), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, primary rotamer), 2.29 (m, 1H, primary rotamer), 2.15 (m, 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 174.0, 173.8, 173.4, 172.9, 168.2.

MS (m / z) 481.2 (M + 1) ⁺ .

Example 18

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) (R) CH (OH) C (O) -Aze-Pab (OMe)

To a solution of Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) OH (0.30 g; 1.7 mmol; see example 17 (5) above (compound ( b))) in DMF (15 ml) under nitrogen atmosphere, H-Aze-Pab (OMe) × 2HCl (0.49 g; 1.5 mmol), RuBOP (0.65 g; 1.2 mmol) and DIPEA were added (0.36 g; 2.8 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo, the residue was chromatographed three times on silica gel, eluting first with CHCl ₃ / EtOH (10: 1), then EtOAc / EtOH ( 10: 1) and finally CHCl ₃ / MeOH (20: 1), which allowed to obtain the title compound (0.47 g; 81%) as a crushable white foam.

Mp 65-75 ° C.

R _f = 0.37 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.58-7.60 (m, 2H), 7.32-7.35 (m, 2H), 7.09-7.12 (m, 1H), 6.96-7.02 (m, 2H ), 6.16 (t, J = 55 Hz, 1H), 5.08-5.18 and 4.74-4.80 (m, 2H), 3.96-4.50 (m, 6H), 3.80 (s, 3H), 2.10-2.75 (m, 2H )

APCl-MS: (M + 1) = 511 m / z.

Example 19

Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) Ph (3-Cl) (5-TMSO) - (R, S) CH (OTMS) CN

To a solution of 3-chloro-5-hydroxybenzaldehyde (9.8 g; 62.6 mmol; see Example 1 (2) above) and Znl ₂ (5.0 g; 15.7 mmol) in anhydrous CH ₂ Cl ₂ (500 ml) trimethylsilyl cyanide (13.7 g; 138 mmol) was added at 0 ° C. The reaction mixture was allowed to warm to room temperature and was stirred overnight. Water (250 ml) was added and the layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (2 × 300 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (16.9 g; 83%) as a yellow oil, which was used without further purification.

R _f = 0.42 (Hex / EtOAc (3: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.06 (s, 1H), 6.86 (s, 2H), 5.40 (s, 1H), 0.30 (s, 9H), 0.24 (s, 9H).

(2) Ph (3-Cl) (5-OH) - (R, S) CH (OH) C (O) OH

A solution of Ph (3-Cl) (5-OTMS) - (R, S) CH (OTMS) CN (22.6 g; 68.8 mmol; see step (1) above) in concentrated HCl (200 ml) was boiled with reflux in nitrogen atmosphere for 3 hours. The reaction mixture was cooled to 0 ° C and slowly alkalinized 2 N. NaOH. The mixture was washed with Et ₂ O (3 × 100 ml) to remove organic impurities. The aqueous layer was acidified with 2 N. HCl and was extracted with EtOAc (3 × 200 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (9.3 g; 67%) as a brown oil, which was used without further purification.

R _f = 0.23 (CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.05 (s, 1H), 6.94 (s, 1H), 6.73 (s, 1H), 5.03 (s, 1H).

(3) Ph (3-Cl) (5-OH) - (R, S) CH (OH) C (O) OEt

To a solution of Ph (3-Cl) (5-OH) - (R, S) CH (OH) C (O) OH (9.3 g; 46.0 mmol; see step (2) above) in absolute EtOH ( 200 ml) concentrated sulfuric acid (0.25 ml) was added and the reaction mixture was refluxed under nitrogen for 4 hours. The reaction mixture was cooled to 0 ° C. and solid NaHCO ₃ (0.2 g) was added. The reaction mixture was concentrated in vacuo and partitioned between saturated NaHCO ₃ (100 ml) and Et ₂ O (3 × 50 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (6.9 g; 65%) as a yellow oil, which was used without further purification.

R _f = 0.62 (CHCl ₃ / MeOH / concentrated NH ₄ OH (6: 3: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 6.99 (s, 1H), 6.81 (s, 2H), 5.07 (s, 1H), 4.16-4.32 (m, 2H), 1.23 (t, J = 7 Hz, 3H).

(4) Ph (3-Cl) (5-OCH ₂ F) - (R, S) CH (OH) C (O) OEt

To a solution of Ph (3-Cl) (5-OH) - (R, S) CH (OH) C (O) OEt (6.1 g; 26.8 mmol; see step (3) above) in DMF (100 ml) in a sealed flask in a nitrogen atmosphere at 0 ° C was added cesium carbonate (13.1 g; 40.2 mmol). The reaction mixture was stirred at 0 ° C for 15 minutes, followed by the addition of potassium iodide (0.5 g; 2.7 mmol). The reaction mixture was cooled to -78 ° C and the vessel was bubbled with chlorofluoromethane (18.4 g; 268 mmol). The sealed flask was then allowed to warm to room temperature and stirred for 18 hours. The reaction mixture was cooled to 0 ° C., carefully vented to remove any excess chlorofluoromethane and partitioned between H ₂ O (20 ml) and Et ₂ O (3 × 50 ml). The combined organic phases were washed with brine (2 × 50 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (gradient from 9: 1 to 3: 1) afforded the subtitled compound (2.4 g; 35%) as a light yellow oil. Note: this compound is poorly distinguishable in UV on TLC. It can be made visible by staining the TLC plate with bromocresol green.

R _f = 0.46 (Hex / EtOAc (2: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.21 (s, 1H), 7.08 (s, 1H), 7.05 (s, 1H), 5.70 (d, J _HF = 54 Hz, 2H), 5.12 (d, J = 5 Hz, 1H), 3.80-4.35 (m, 2H), 3.50 (d, J = 5 Hz, 1H), 1.26 (t, J = 7 Hz, 3H).

(5) Ph (3-Cl (5-OCH ₂ F) - (R, S) CH (OH) C (O) OH

To a solution of Ph (3-Cl) (5-OCH ₂ F) - (R, S) CH (OH) C (O) OEt (1.8 g; 6.8 mmol; see step (4) above) in a mixture H ₂ O / THF (30 ml, 1: 2) at 0 ° C. under a nitrogen atmosphere was added lithium hydroxide monohydrate (0.40 g; 10.3 mmol). The mixture was stirred at 0 ° C for 2 hours. The reaction mixture was concentrated in vacuo and partitioned between H ₂ O (5 ml) and Et ₂ O (2 × 20 ml). The aqueous layer was carefully acidified with 0.2 N HCl at 0 ° C and was extracted with EtOAc (3 × 30 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (1.4 g; 87%) as a colorless oil, which solidified upon standing to a white solid.

R _f = 0.43 (CHCl ₃ / MeOH / Et ₃ N (6: 2: 1)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.24 (s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 5.78 (d, J _HF = 54 Hz, 2H), 5.13 (s, 1H).

(6) Ph (3-Cl (5-OCH ₂ F) - (R) CH (OH) C (O) OH (a) and Ph (3-Cl) (5-OCH ₂ F) - (S) CH (OAc) C (O) OH (b)

Mixture of Ph (3-Cl) (5-OCH ₂ F) - (R, S) CH (OH) C (O) OH (3.2 g; 13.9 mmol; see step (5) above) and Lipase PS Atapo (1.9 g) in vinyl acetate (150 ml) and MTBE (150 ml) were heated at 70 ° C in a nitrogen atmosphere for 3 days. The reaction mixture was cooled, filtered through Celite® and the filter cake was washed with EtOAc. The filtrate was concentrated in vacuo and flash chromatographed on silica gel eluting with CHCl ₃ / MeOH / Et ₃ N (15: 1: 0.5), which afforded the triethylamine salt of the subtitle compound (a) (0.50 g; 21%), which was used without neutralization. In addition, a triethylamine salt of the subtitle compound (b) was obtained (0.46 g; 20%).

Data for the subtitled compound (a)

R _f = 0.19 (CHCl ₃ / MeOH / Et ₃ N (15: 1: 0.5)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.26 (s, 1H), 7.18 (s, 1H), 6.97 (s, 1H), 5.74 (d, J _HF = 54 Hz, 2H), 4.81 (s, 1H), 3.17 (q, J = 7 Hz, 6H), 1.28 (t, J = 7 Hz, 9H).

Data for the subtitle compound (b)

R _f = 0.33 (CHCl ₃ / MeOH / Et ₃ N (15: 1: 0.5)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.28 (s, 1H), 7.19 (s, 1H), 7.09 (s, 1H), 5.76 (d, J _HF = 54 Hz, 2H), 5.75 (s, 1H), 3.17 (q, J = 7 Hz, 6H), 2.16 (s, 3H), 1.28 (t, J = 7 Hz, 9H).

(7) Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of the triethylamine salt Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) OH (0.50 g; 1.50 mmol; see step (6) above) and HAze -Pab (Teoc) · HCl (0.87 g; 1.90 mmol) in anhydrous DMF (15 ml) in a nitrogen atmosphere at 0 ° C were added RuBOP (0.85 g; 2.60 mmol) and DIPEA (0, 48 g; 3.70 mmol). The reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction mixture was concentrated in vacuo and flash chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (9: 1) and a second time with EtOAc / EtOH (20: 1), which afforded the subtitled compound (0.23 g ; 26%) as crushable white foam.

Mp 88-92 ° C.

R _f = 0.61 (CHCl ₃ / EtOH (9: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.81 (d, J = 8 Hz, 2H), 7.40-7.42 (m, 2H), 7.06-7.23 (m, 3H), 5.76 (d, J _HF = 51 Hz, 2H), 5.10-5.16 and 4.77-4.83 (m, 2H), 3.80-4.49 (m, 6H), 2.30-2.53 (m, 2H), 1.08 (t, J = 7 Hz, 2H ), 0.08 (s, 9H).

APCl-MS: (M + 1) = 593 m / z.

(8) Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.051 g; 0.086 mmol; see step (7) above) was dissolved in 3 ml TFA and allowed to interact for 20 minutes. TFA was evaporated and the residue was dried by freezing from a mixture of water / acetonitrile. The purity of the product was 95% with 5% defluoromethylated material. Attempts to clean it using preparative OFLC (CH ₃ CN / 0.1 M NH ₄ OAc) were unsuccessful, and the material, partially in the form of acetate, was dissolved in 5 ml of TFA, evaporated and freeze-dried to obtain 26 mg (51%) the title compound as its TFA salt. Purity 95%.

¹ H-NMR (600 MHz; CD ₃ OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.21 (s, 1H, main rotamer), 7.17 (s, 1H, minor rotamer), 7.13 (s, 1H, main rotamer), 7.09 (s, 1H, minor rotamer), 7.07 (m, 1H, main rotamer), 7.04 (m, 1H, minor rotamer), 5.73 (d, 2H), 5.18 (m, 1H, minor rotamer), 5.16 (s, 1H, primary rotamer), 5.09 (s, 1H, minor rotamer), 4.78 (m, 1H, minor rotamer), 4.56 (d, 1H, main rotamer), 4.50 (d, 1H, minor rotamer), 4.46 (d, 1H, minor rotamer), 4.45 (d, 1H, main rotamer), 4.35 (m, 1H, main rotamer), 4.14 (m, 1H, main rotamer), 4.05 (m, 1H, minor rotamer), 3.97 (m, 1H, minor rotamer), 2.68 (m, 1H, minor rotamer), 2.52 (m, 1H, main rotamer), 2.28 (m, 1H, main rotamer), 2.19 (m, 1H, minor rotamer).

¹³ C-NMR (150 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 173.9, 173.3, 172.9, 168.2.

ESI-MS +: (M + 1) = 449 (m / z).

Example 20

Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OMe)

To a solution of the triethylamine salt Ph (3-Cl) (5-OCH ₂ F) - (R) CH (OH) C (O) OH (0.60 g; 1.80 mmol; see Example 19 (6) above) and HAze-Pab (OMe) · 2HCl (0.79 g; 2.30 mmol) in DMF (15 ml) in a nitrogen atmosphere at 0 ° C was added RuBOP (1.04 g; 1.90 mmol) and DIPEA (0, 58 g; 4.50 mmol). The reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction mixture was concentrated in vacuo and flash chromatographed on silica gel three times, eluting first with CHCl ₃ / EtOH (9: 1) and then twice with EtOAc / EtOH (20: 1) to give the title compound (0.22 g ; 26%) as crushable white foam.

Mp 66-70 ° C.

R _f = 0.45 (CHCl ₃ / EtOH (9: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.59 (d, J = 8 Hz, 2H), 7.32 (d, J = 7 Hz, 2H), 7.06-7.23 (m, 3H), 5.75 (s, J _HF = 54 Hz, 1H), 5.10-5.16 and 4.78-4.84 (m, 2H), 4.11-4.45 (m, 4H), 3.80 (s, 3H), 2.10-2.75 (m, 2H).

¹³ C-NMR (150 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 173.0, 170.8, 170.7, 152.5.

APCl-MS: (M + 1) = 479 m / z.

Example 21

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) (2-Monofluoroethyl) methanesulfonate

To a stirred solution of 2-fluoroethanol (5.0 g; 78.0 mmol) in CH ₂ Cl ₂ (90 ml) under nitrogen atmosphere at 0 ° C was added triethylamine (23.7 g; 234 mmol) and methanesulfonyl chloride ( 10.7 g; 93.7 mmol). The mixture was stirred at 0 ° C for 1.5 h, diluted with CH ₂ Cl ₂ (100 ml) and washed with 2 N. HCl (100 ml). The aqueous layer was extracted with CH ₂ Cl ₂ (50 ml), the combined organic extracts were washed with brine (75 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (9.7 g; 88%) as a yellow oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 4.76 (t, J = 4 Hz, 1H), 4.64 (t, J = 4 Hz, 1H), 4.52 (t, J = 4 Hz, 1H), 4.43 (t , J = 4 Hz, 1H), 3.09 (s, 3H).

(2) 3-Chloro-5-monofluoroethoxybenzaldehyde

To a solution of 3-chloro-5-hydroxybenzaldehyde (8.2 g; 52.5 mmol; see example 1 (2) above) and potassium carbonate (9.4 g; 68.2 mmol) in DMF (10 ml) in the atmosphere nitrogen was added dropwise a solution of (2-monofluoroethyl) methanesulfonate (9.7 g; 68.2 mmol; see step (1) above) in DMF (120 ml) at room temperature. The mixture was heated to 100 ° C. for 5 hours, then stirred overnight at room temperature. The reaction mixture was cooled to 0 ° C, poured into chilled in ice 2 N. HCl and extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with a Hex / EtOAc mixture (4: 1), which afforded the subtitled compound (7.6 g; 71%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.92 (s, 1H), 7.48 (s, 1H), 7.32 (s, 1H), 7.21 (s, 1H), 4.87 (t, J = 4 Hz, 1H) 4.71 (t, J = 3 Hz, 1H), 4.33 (t, J = 3 Hz, 1H), 4.24 (t, J = 3 Hz, 1H).

(3) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R, S) CH (OTMS) CN

To a solution of 3-chloro-5-monofluoroethoxybenzaldehyde (7.6 g; 37.5 mmol; see step (2) above) and zinc iodide (3.0 g; 9.38 mmol) in CH ₂ Cl ₂ (310 ml) trimethylsilyl cyanide (7.4 g; 75.0 mmol) was added dropwise at 0 ° C. under a nitrogen atmosphere. The mixture was stirred at 0 ° C. for 3 hours and at room temperature overnight. The reaction mixture was diluted with H ₂ O (300 ml), the organic layer was separated, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (10.6 g; 94%) as a brown oil, which was used without further purification or characterization.

(4) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R, S) CH (OH) C (O) OH

Concentrated hydrochloric acid (100 ml) was added to Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R, S) CH (OTMS) CN (10.6 g; 5.8 mmol; see step (3) ) above), and the solution was stirred at 100 ° C for 3 h. After cooling to room temperature, the reaction mixture was further cooled to 0 ° C, slowly made alkaline with 3 N NaOH (~ 300 ml) and washed with Et ₂ O (3 × 200 ml). The aqueous layer was acidified with 2 N. HCl (80 ml) and extracted with EtOAc (3 × 300 ml). The combined EtOAc extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (8.6 g; 98%) as a pale yellow solid that was used without further purification.

R _f = 0.28 (CHCl ₃ / MeOH / concentrated NH ₄ OH (90: 8: 2)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77-4.81 (m, 1H), 4.62 -4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).

(5) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (S) CH (OAc) C (O) OH (a) and Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) OH (b)

Solution Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R, S) CH (OH) C (O) OH (8.6 g; 34.5 mmol; see step (4) above) and Lipases PS "Amano" (4.0 g) in vinyl acetate (250 ml) and MTBE (250 ml) were heated at 70 ° C in a nitrogen atmosphere for 3 days. The reaction mixture was cooled to room temperature and the enzyme was removed by filtration through Celite®. The filter cake was washed with EtOAc and the filtrate was concentrated in vacuo. Silica gel chromatography, eluting with CHCl ₃ / MeOH / Et ₃ N (90: 8: 2), afforded the triethylamine salt of the subtitle compound (a) as a yellow oil. In addition, a triethylamine salt of the subtitle compound (b) (4.0 g) was obtained. The salt of the subtitle compound (b) was dissolved in H ₂ O (250 ml), acidified with 2 N. HCl and was extracted with EtOAc (3 × 200 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitle compound (b) (2.8 g; 32%) as a yellow oil.

Data for the subtitle compound (b):

R _f = 0.28 (CHCl ₃ / MeOH / concentrated NH ₄ OH (90: 8: 2)).

¹ H NMR (300 MHz; CD ₃ OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77-4.81 (m, 1H), 4.62 -4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).

(6) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) OH (940 mg; 3.78 mmol; see step (5) above) in DMF (30 ml) under a nitrogen atmosphere at 0 ° C, HAze-Pab (Teoc) · HCl (2.21 g; 4.91 mmol), RuBOP (2.16 g; 4.15 mmol) and DIPEA (1.22 g; 9.45 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature for 4 hours. The mixture was concentrated in vacuo and the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1), which made it possible to obtain the compound indicated in the subtitle (450 mg; 20%) as a crushable white foam.

Mp 80-88 ° C.

R _f = 0.60 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CO ₃ CD; complex mixture of rotamers) δ 7.79 (d, J = 8 Hz, 2H), 7.42 (d, J = 8 Hz, 2H), 7.05-7.08 (m, 1H), 6.93 -6.99 (m, 2H), 5.08-5.13 (m, 1H), 4.75-4.80 (m, 2H), 4.60-4.68 (m, 1H), 3.95-4.55 (m, 8H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

APCl-MS: (M + 1) = 607 m / z.

(7) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.357 g; 0.589 mmol; see step (6) above) was dissolved in 10 ml of TFA and allowed to react for 40 minutes. TFA was evaporated and the residue was freeze-dried from water / acetonitrile to give 0.33 g (93%) of the title compound as its TFA salt.

¹ H-NMR (600 MHz; CD ₃ OD, rotamers): δ 7.8-7.7 (m, 2H), 7.54 (d, 2H), 7.08 (s, 1H, main rotamer), 7.04 (s, 1H, minor rotamer ), 6.99 (s, 1H, main rotamer), 6.95 (s, 1H), 6.92 (s, 1H, minor rotamer), 5.18 (m, 1H, minor rotamer), 5.14 (s, 1H, main rotamer), 5.08 (s, 1H, minor rotamer), 4.80 (m, 1H, main rotamer), 4.73 (m, 1H), 4.65 (m, 1H), 4.6-4.4 (m, 2H), 4.35 (m, 1H, main rotamer ), 4.21 (doublet of multiplets, 2H), 4.12 (m, 1H, main rotamer), 4.06 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.69 (m, 1H, minor rotamer), 2.53 (m, 1H, main rotamer), 2.29 (m, 1H, main rotamer), 2.14 (m, 1H, minor rotamer).

¹³ C-NMR (150 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 172.8, 172.1, 167.4.

ESI-MS +: (M + 1) = 463 (m / z).

Example 22

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OMe)

To a solution of Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) OH (818 mg; 3.29 mmol; see Example 21 (5) above) in DMF ( 30 ml) under a nitrogen atmosphere at 0 ° C, HAze-Pab (OMe) · 2HCI (1.43 g; 4.27 mmol), RuBOP (1.89 g; 3.68 mmol) and DIPEA (1.06 g) were added ; 8.23 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo, and the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1) to give the title compound (880 mg; 54%) in the form of crushed white foam.

Mp 65-72 ° C.

R _f = 0.60 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CO ₃ CD; complex mixture of rotamers) δ 7.58-7.60 (d, J = 8 Hz, 2H), 7.34 (d, J = 7 Hz, 2H), 7.05-7.08 (m, 2H) , 6.95-6.99 (m, 1H), 5.08-5.13 (m, 1H), 4.77-4.82 (m, 1H), 4.60-4.68 (m, 1H), 3.99-4.51 (m, 7H), 3.82 (s, 3H), 2.10-2.75 (m, 2H).

¹³ C-NMR (150 MHz; CD ₃ CD) (carbonyl and / or amidine carbons) δ 173.3, 170.8, 152.5.

APCI-MS: (M + 1) = 493 m / z.

Example 23

Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 1,3-Difluoroisopropylmethanesulfonate

To a solution of 1,3-difluoro-2-propanol (7.0 g; 72.8 mmol) in CH ₂ Cl ₂ (100 ml) stirred under a magnetic stirrer in a nitrogen atmosphere at 0 ° C was added triethylamine (22.1 g; 219 mmol) and methanesulfonyl chloride (10.0 g; 87.4 mmol). The mixture was stirred at 0 ° C. for 3 hours. The mixture was washed with 2 N. HCl (150 ml) and the layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (200 ml), the combined organic extracts were washed with brine (100 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (11.5 g; 91%) as a yellow oil, which is used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 4.97-5.08 (m, 1H), 4.75-4.77 (m, 2H), 4.59-4.61 (m, 2H), 3.12 (s, 3H).

(2) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) CHO

To a solution of 3-chloro-5-hydroxybenzaldehyde (8.0 g; 50.7 mmol; see example 1 (2) above) and potassium carbonate (9.1 g; 66.0 mmol) in DMF (75 ml) in the atmosphere nitrogen was added dropwise a solution of 1,3-difluoroisopropylmethanesulfonate (11.5 g; 66.0 mmol; see step (1) above) in DMF (75 ml) at room temperature. The mixture was heated to 110 ° C for 18 hours. The reaction mixture was cooled to 0 ° C, poured into 2N chilled in ice. HCl (200 ml) and was extracted with EtOAc (3 × 250 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with a Hex / EtOAc mixture (4: 1), which afforded the subtitled compound (4.4 g; 37%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.92 (s, 1H), 7.51 (s, 1H), 7.36 (s, 1H), 7.26 (s, 1H), 4.70-4.89 (m, 3H), 4.63- 4.68 (m, 2H).

(3) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R, S) CH (OTMS) CN

To a solution of Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) CHO (4.4 g; 18.7 mmol; see step (2) above) and zinc iodide (1.5 g; 4.67 mmol) in CH ₂ Cl ₂ (200 ml) at 0 ° C under nitrogen atmosphere was added dropwise trimethylsilyl cyanide (3.7 g; 37.3 mmol). The mixture was stirred at 0 ° C. for 3 hours and at room temperature overnight, then diluted with H ₂ O (200 ml). The organic layer was separated, dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (5.5 g; 87%) as a brown oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.12 (s, 1H), 7.00 (s, 2H), 5.42 (s, 1H), 4.70-4.80 (m, 3H), 4.59-4.64 (m, 2H), 0.26 (s, 9H).

(4) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R, S) CH (OH) C (O) OH

Concentrated hydrochloric acid (50 ml) was added to Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R, S) CH (OTMS) CN (5.5 g; 16.3 mmol; see step (3) above), and the solution was stirred at 100 ° C. for 1.5 hours. After cooling to room temperature, the reaction mixture was further cooled to 0 ° C., slowly made basic with 3 N NaOH (~ 200 ml) and washed with Et ₂ O (3 × 200 ml). The aqueous layer was acidified with 2 N. HCl (75 ml) and was extracted with EtOAc (3 × 200 ml). The combined EtOAc extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitled compound (4.6 g; 100%) as a brown oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.14 (s, 1H), 7.08 (s, 1H), 7.02 (s, 1H), 5.12 (s, 1H), 4.70-4.90 (m, 3H), 4.52 -4.67 (m, 2H).

(5) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (S) CH (OAc) C (O) OH (a) and Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) OH (b)

Solution Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R, S) CH (OH) C (O) OH (4.6 g; 16.4 mmol; see step (4) above) ) and Lipases PS "Amano" (3.0 g) in vinyl acetate (150 ml) and MTBE (150 ml) were heated at 70 ° C in a nitrogen atmosphere for 2.5 days. The reaction mixture was cooled to room temperature, the enzyme was removed by filtration through Celite®. The filter cake was washed with EtOAc and the filtrate was concentrated in vacuo. Silica gel chromatography, eluting with CHCl ₃ / MeOH / Et ₃ N (90: 8: 2), afforded the triethylamine salt of the subtitle compound (a) as a yellow oil. In addition, a triethylamine salt of the subtitle compound (b) (2.2 g) was obtained, this salt was dissolved in H ₂ O (100 ml), acidified with 2 N HCl and was extracted with EtOAc (3 × 200 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give the subtitle compound (b) (1.4 g; 29%) as a yellow oil.

Data for the subtitle compound (b)

¹ H NMR (300 MHz; CD ₃ OD) δ 7.14 (s, 1H), 7.08 (s, 1H), 7.02 (s, 1H), 5.12 (s, 1H), 4.70-4.90 (m, 3H), 4.52 -4.67 (m, 2H).

(6) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) OH (824 mg; 2.94 mmol; see step (5) above) in DMF (30 ml) under a nitrogen atmosphere at 0 ° C, HAze-Pab (Teoc) · HCl (1.71 g; 3.81 mmol), RuBOP (1.68 g; 3.23 mmol) and DIPEA (949 mg; 7.34 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo, the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1), which afforded the subtitled compound (720 mg; 38%) in crushed white foam.

Mp 78-84 ° C.

R _f = 0.62 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ CD; complex mixture of rotamers) δ 7.79 (d, J = 8 Hz, 2H), 7.42 (d, J = 8 Hz, 2H), 7.00-7.12 (m, 3H), 5.08 -5.20 (m, 1H), 3.97-4.80 (m, 12H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 639 m / z.

(7) Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.129 g; 0.202 mmol; see step (6) above) dissolved in 3 ml of TFA and allowed to react for 20 minutes. TFA was evaporated and the residue was freeze-dried from water / acetonitrile to give 0.123 g (100%) of the title compound as its TFA salt.

¹ H-NMR (400 MHz; CD ₃ CD) rotamers: δ 7.8-7.7 (m, 2H), 7.55 (d, 2H), 7.2-7.0 (m, 3H), 5.18 (m, 1H, minor rotamer), 5.15 (s, 1H, main rotamer), 5.08 (s, 1H, minor rotamer), 4.80 (m, 1H, main rotamer partially covered by the peak of CD ₃ OH), 4.75-4.4 (m, 7H), 4.38 (m, 1H, main rotamer), 4.15 (m, 1H, main rotamer), 4.1-3.9 (m, 2H, 2 signals from the minor rotamer), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, main rotamer) 2.30 (m, 1H, main rotamer); 2.15 (m, 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.9, 172.6, 172.2, 171.7, 167.1.

ESI-MS +: (M + 1) = 495 (m / z).

Example 24

Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) Aze-Pab (OMe)

To a solution of Ph (3-Cl) (5-OCH (CH ₂ F) ₂ ) - (R) CH (OH) C (O) OH (513 mg; 1.83 mmol; see Example 23 (5) above) in DMF (30 ml) in a nitrogen atmosphere at 0 ° C was added HAze-Pab (OMe) · 2HCl (797 mg; 2.38 mmol), RuBOP (1.04 g; 2.01 mmol) and DIPEA (591 mg; 4 , 57 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo, the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1), which afforded the title compound (370 mg; 39%) in crushed white foam.

Mp 58-63 ° C.

R _f = 0.66 (CHCl ₃ / EtOH (10: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.58-7.60 (d, J = 8 Hz, 2H), 7.34 (d, J = 8 Hz, 2H), 7.00-7.12 (m, 3H) 5.08-5.20 (m, 1H), 4.65-4.82 (m, 3H), 4.28-4.65 (m, 5H), 3.92-4.18 (m, 2H), 3.82 (s, 3H), 2.10-2.75 (m, 2H).

¹³ C-NMR (150 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 173.2, 170.8, 152.5.

APCI-MS: (M + 1) = 525 m / z.

Example 25

Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 1-Bromo-3-fluoro-5-benzyloxybenzene

Sodium hydride (60% dispersion in oil; 24.0 g, 0.48 mol) was added portionwise to a stirred solution of anhydrous benzyl alcohol (64.5 g; 0.60 mol) in THF (1.0 L). After the mixture was stirred for 1 hour, a solution of 1-bromo-3,5-difluorobenzene (76.8 g; 0.40 mol) in THF (100 ml) was added dropwise over a period of 1 hour. The reaction mixture was stirred at room temperature within 2 days. Water (400 ml) was added and THF was removed in vacuo. The aqueous layer was extracted with hexane (3 × 150 ml). The combined organic extracts were washed with 2 N. NaOH (2 × 100 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (110.7 g; 98%) as a light yellow oil, which was used without further cleaning up.

R _f = 0.47 (Hex).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.36-7.41 (m, 5H), 6.94 (bs, 1H), 6.87 (d, J _HF = 8 Hz, 1H), 6.63 (d, J _HF = 10 Hz, 1H), 5.03 (s, 2H).

(2) 3-Bromo-5-fluorophenol

To a solution of 1-bromo-3-fluoro-5-benzyloxybenzene (110.0 g; 0.39 mol; see step (1) above) and N, N-dimethaniline (474.0 g; 3.92 mol) in anhydrous CH ₂ Cl ₂ (1.0 L) was added at 0 ° C. aluminum chloride (156.0 g; 1.17 mol). After 10 minutes, the ice bath was removed and stirring was continued for 2 hours. The reaction was quenched by carefully adding 3N. HCl (600 ml). The layers were separated and the aqueous layer was extracted with CH ₂ Cl ₂ (2 × 150 ml). The combined organic extracts were washed with 2 N. HCl (250 ml) and H ₂ O (3 × 250 ml). 15% KOH (500 ml) was added to the organic layer and the layers were separated. The organic layer was extracted again with 2 N. HCl (2 × 70 ml). The combined aqueous layers were washed with CH ₂ Cl ₂ (3 × 100 ml) and then acidified with 4 N. HCl. Then the aqueous layer was extracted with Et ₂ O (3 × 125 ml), the combined Et ₂ O extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (69.0 g; 92%) in the form of a brown oil, which was used without further purification.

Mp 33-35 ° C.

R _f = 0.25 (CHCl ₃ ).

¹ H NMR (300 MHz; DMSO-d ₆ ) δ 10.38 (s, 1H), 6.90 (dd, J _HF = 11 Hz, J = 2 Hz, 1H), 6.81 (s, 1H), 6.59 (dt, J _HF = 11 Hz, J = 2 Hz, 1H).

APCI-MS: (M-1) = 189 m / z.

(3) 1-Bromo-3-fluoro-5-difluoromethoxybenzene

A mixture of 3-bromo-5-fluorophenol (6.1 g; 31.0 mmol; see step (2) above) and chlorodifluoromethane (13.0 g; 150.0 mmol) in iso-PrOH (100 ml) and 30% -nom KOH (80 ml) was heated in a hermetically sealed flask for 18 h at 80-85 ° С. The reaction mixture was cooled to room temperature and the layers were separated. The organic layer was concentrated in vacuo, resulting in a colorless oil. The aqueous layer was extracted with Et ₂ O (3 × 30 ml). The crude oil and the combined organic extracts were washed with 2 N. NaOH (3 × 30 ml) and H ₂ O (3 × 30 ml). The organic phase was then dried (Na ₂ SO ₄ ), filtered through a small pad of silica gel and concentrated in vacuo, which afforded the subtitle compound (6.1 g; 79%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.11-7.14 (m, 2H), 6.84 (dt, J = 9 Hz, J = 2 Hz, 1H), 6.50 (t, J _HF = 72 Hz, 1H).

(4) 1-Fluoro-3-difluoromethoxy-5-vinylbenzene

Tri (butyl) vinyl stannane (7.0 g; 22.2 mmol) was added to a suspension of 1-bromo-3-fluoro-5-difluoromethoxybenzene (4.9 g; 20.2 mmol; see step (3) above), dichlorobis (triphenylphosphine) palladium (II) (1.42 g; 2.02 mmol) and anhydrous lithium chloride (0.90 g; 20.2 mmol) in THF (40 ml) under nitrogen at 65 ° C and the mixture was stirred in for 5 hours. The reaction mixture was cooled to 0 ° C and 1 N was added. NaOH (90 ml). The biphasic mixture was vigorously stirred for 1 hour, then the layers were separated. The aqueous layer was extracted with Et ₂ O (3 × 70 ml). The combined organic layers were washed with 2 N. NaOH (2 × 40 ml) and H ₂ O (40 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexane afforded the subtitled compound (2.2 g; 57%) as a colorless oil.

R _f = 0.47 (Hex).

¹ H NMR (300 MHz; CDCl ₃ ) δ 6.93-6.99 (m, 2H), 6.73-6.78 (m, 1H), 6.67 (dd, J = 18 Hz, J = 11 Hz, 1H), 6.51 (t, J _HF = 73 Hz, 1H), 5.77 (d, J = 18 Hz, 1H), 5.36 (d, J = 11 Hz, 1H).

(5) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OH

2-Methyl-2-propanol (140 ml), H ₂ O (140 ml) and AD-mix-β (39.2 g) were combined and cooled to 0 ° C. 1-Fluoro-3-difluoromethoxy-5-vinylbenzene (5.0 g; 26.4 mmol; see step (4) above) was immediately dissolved in a small amount of 2-methyl-2-propanol and the heterogeneous mixture was vigorously stirred at 0 ° C. until TLC showed a lack of starting material. The reaction was quenched at 0 ° C by adding sodium sulfite (42.0 g) and then warmed to room temperature and stirred for 60 minutes. The reaction mixture was extracted with Et ₂ O (3 × 120 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel, eluting with CHCl ₃ / EtOAc (3: 2), afforded the subtitled compound (5.8 g; 98%) as a colorless oil.

R _f = 0.41 (CHCl ₃ / EtOAc (3: 2)).

¹ H NMR (300 MHz; COCl ₃ ) δ 6.96-6.99 (m, 2H), 6.77-6.82 (m, 1H), 6.51 (t, J _HF = 73 Hz, 1H), 4.79-4.85 (m, 1H) , 3.76-3.84 (m, 1H), 3.58-3.66 (m, 1H), 2.66 (d, J = 3 Hz, 1H), 2.00 (t, J = 6 Hz, 1H).

HPLC analysis: 89.2%,> 99% e.e., ChiralPak AD column (Hex / EtOH mobile phase (95: 5)).

(6) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OTBS

Solution Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OH (5.5 g; 24.7 mmol; see step (5) above), 4- (dimethylamino) pyridine ( 121 mg; 1.0 mmol) and triethylamine (3.0 g; 29.6 mmol) in anhydrous CH ₂ Cl ₂ (100 ml) were cooled to 0 ° C. A 1.0 M solution of tert-butyldimethylsilyl chloride in CH ₂ Cl ₂ (26.0 ml; 26.0 mmol) was added dropwise, the reaction mixture was allowed to warm to room temperature and was stirred overnight. A saturated solution of ammonium chloride (60 ml) was added and the layers were separated. The organic layer was washed with a saturated solution of ammonium chloride (60 ml) and H ₂ O (2 × 35 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel, eluting with CHCl ₃ / Hex (3: 1), afforded the subtitled compound (7.9 g; 85%) as a yellow oil.

R _f = 0.47 (CHCl ₃ / Hex (3: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 6.95-6.98 (m, 2H), 6.76-6.79 (m, 1H), 6.51 (t, J _HF = 73 Hz, 1H), 4.71-4.74 (m, 1H) , 3.75-3.80 (m, 1H), 3.48-3.54 (m, 1H), 2.99 (bs, 1H), 0.91 (s, 9H), 0.05 (s, 3H), 0.00 (s, 3H).

(7) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OTBS

To a solution of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OTBS (7.9 g; 0.51 mmol; see step (6) above) and DIPEA (4.9 g ; 48.1 mmol) in anhydrous CH ₂ Cl ₂ (50 ml) at 0 ° C. under nitrogen atmosphere, 2-methoxyethoxymethyl chloride (6.6 g; 48.1 mmol) was added dropwise. The mixture was stirred for 24 hours. A saturated solution of ammonium chloride (70 ml) was added and the layers were separated.

The organic layer was washed with a saturated solution of ammonium chloride (70 ml) and H ₂ O (3 × 60 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (8.8 g; 99%) as a yellow oil, which was used without further purification.

R _f = 0.41 (CHCl ₃ / EtOAc (4: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.20 (s, 1H), 7.06 (s, 1H), 7.02 (s, 1H), 6.50 (t, J _HF = 73 Hz, 1H), 4.79-4.81 (m, 1H), 4.66-4.68 (m, 2H), 3.47-3.82 (m, 6H), 3.36 (s, 3H), 0.85 (s, 9H), 0.01 (s, 3H), 0.00 (s, 3H) )

(8) Ph (3-F) (5-OCHF ₂ ) (R) CH (OMEM) CH ₂ OH

To a solution of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OTBS (9.3 g; 21.9 mmol; see step (7) above) in THF (60 ml) at at room temperature, a 1.0 M solution of tetrabutylammonium fluoride in THF (70.0 ml; 70.0 mmol) was added and the mixture was stirred overnight under nitrogen. The reaction mixture was concentrated in vacuo. The yellow residue was dissolved in Et ₂ O (100 ml) and hexane (100 ml) and washed successively with a saturated solution of ammonium chloride (2 × 150 ml) and H ₂ O (3 × 70 ml). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (1: 1) afforded the subtitled compound (4.2 g; 62%) as a yellow oil.

R _f = 0.42 (Hex / EtOAc (1: 1)).

¹ H NMR (300 MHz; CDCl ₃ ) δ 6.91-6.95 (m, 2H), 6.75-6.81 (m, 1H), 6.51 (t, J _HF = 73 Hz, 1H), 4.80-4.82 (m, 1H) , 4.70-4.74 (m, 2H), 3.88-3.93 (m, 1H), 3.67-3.71 (m, 3H), 3.53-3.56 (m, 2H), 3.39 (s, 3H), 2.96-2.99 (m, 1H).

(9) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH

A solution of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OH (4.2 g; 13.4 mmol; see step (8) above) in acetone (100 ml) was added to aqueous 5% solution of NaHCO ₃ (35 ml). This heterogeneous mixture stirred on a magnetic stirrer was cooled to 0 ° C and potassium bromide (159 mg; 1.3 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical (2.2 g; 14, 1 mmol). Then, sodium hypochlorite (5.25%; 30 ml) was added dropwise over a period of 20 minutes while stirring the mixture vigorously and maintaining the temperature at 0 ° C. After 1 h, additional sodium hypochlorite (30 ml) and a 5% NaHCO ₃ solution (35 ml) were added, stirring was continued at 0 ° C for 2 h. Acetone was removed in vacuo. The aqueous layer was washed with Et ₂ O (4 × 40 ml). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (4 × 50 ml). Then, the combined EtOAc extracts were washed successively with H ₂ O (4 × 30 ml) and brine (60 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (4.3 g ; 98%) as a colorless oil, which was used without further purification.

R _f = 0.74 (CHCl ₃ / MeOH / Et ₃ N (8.0: 1.5: 0.5)).

¹ H NMR (300 MHz; acetone-d ₆ ) δ 7.16-7.18 (m, 2H), 7.16 (t, J _HF = 89 Hz, 1H), 7.00-7.03 (m, 1H), 5.30 (s, 1H) 4.88 (d, J = 7 Hz, 1H), 4.80 (d, J = 7 Hz, 1H), 3.54-3.75 (m, 2H), 3.46-3.49 (m, 2H), 3.28 (s, 3H).

(10) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH (1.1 g; 3.4 mmol; see step (9) above) in DMF (20 ml ) under a nitrogen atmosphere at 0 ° C, HAze-Pab (Teoc) · HCl (2.0 g; 4.4 mmol), RuBOP (1.9 g; 3.7 mmol) and DIPEA (1.1 g; 8) were added 4 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1), which afforded the subtitled compound (1.3 g; 56% ) in the form of crushed white foam.

R _f = 0.65 (CHCl ₃ / EtOH (15: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.80-7.84 (m, 2H), 7.40-7.46 (m, 2H), 6.95-7.16 (m, 3H), 6.92 and 6.88 (t, J _HF = 73 Hz, 1H), 5.28 and 5.08 (s, 1H), 5.18-5.22 and 4.70-4.78 (m, 1H), 4.50-4.75 (m, 1H), 4.30-4.49 (m, 2H), 4.21- 4.26 (m, 1H), 3.97-4.08 (m, 1H), 3.35-3.72 (m, 6H), 3.30 (s, 3H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

(11) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

Mixture of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc) (590 mg; 0.87 mmol; see step (10) above) and tetrabromide carbon (287 mg; 0.87 mmol) in 2-propanol (20 ml) was refluxed for 1.5 hours. Then this mixture was concentrated in vacuo, partitioned between H ₂ O (50 ml) and EtOAc (3 × 50 ml). The aqueous layer was extracted with additional EtOAc (2 × 10 ml). The combined organic extracts were washed with brine (30 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel, eluting with CHCl ₃ / EtOH (15: 1), afforded the subtitle compound (60 mg; 12%) as a crushable white foam. R _f = 0.46 (CHCl ₃ / EtOH (15: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.74 (d, J = 8 Hz, 2H), 7.35-7.37 (m, 2H), 6.97-7.07 (m, 2H), 6.80-6.84 ( m, 1H), 6.82 and 6.80 (t, J _HF = 73 Hz, 1H), 5.10 and 5.06 (s, 1H), 4.68-4.70 (m, 1H), 3.97-4.60 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 595 m / z.

(12) Ph (3-F) (5-OCHF ₂ ) (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.053 g; 0.089 mmol; see step (11) above) was dissolved in 3 ml of TFA and allowed to interact for 80 minutes while cooling in an ice bath. TFA was evaporated and the residue was freeze dried from a mixture of water / acetonitrile to give 0.042 g (80%) of the title compound as its TFA salt.

¹ H-NMR (300 MHz; CD ₃ OD) rotamers: δ 7.7-7.6 (m, 2H), 7.5-7.4 (m, 2H), 7.1-6.6 (m, 4H), 5.2-5.0 (m, 1H plus minor rotamer 1H), approximately 4.8 (main rotamer of the previous signal, blocked by the signal from CD ₃ OH), 4.6-4.3 (m, 2H), 4.26 (m, 1H, main rotamer), 4.10 (m, 1H, main rotamer), 3.96 (m, 1H, minor rotamer), 3.89 (m, 1H, minor rotamer), 2.60 (m, 1H, minor rotamer), 2.44 (m, 1H, main rotamer), 2.19 (m, 1H, main rotamer), 2.05 (m, 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 172.8, 172.0, 167.0.

ESI-MS +: (M + 1) = 451 (m / z).

Example 26

Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

(1) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (OMe)

To a solution of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH (1.0 g; 3.1 mmol; see Example 25 (9) above) in DMF (30 ml) under a nitrogen atmosphere at 0 ° C, HAze-Pab (OMe) · 2HCl (1.4 g; 4.1 mmol), RuBOP (1.8 g; 3.4 mmol) and DIPEA (1.0 g; 7.8 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo, the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and EtOAc a second time, which afforded the subtitled compound (1.5 g; 79%) as a crushed white foam.

R _f = 0.24 (EtOAc).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.58-7.62 (m, 2H), 7.32-7.38 (m, 2H), 7.03-7.16 (m, 3H), 6.92 and 6.88 (d, J _HF = 73 Hz, 1H), 5.27 and 5.08 (s, 1H), 5.22-5.15 and 4.75-4.80 (m, 1H), 4.38-4.65 (m, 5H), 3.92-4.27 (m, 1H), 3.82 ( s, 3H), 3.43-3.68 (m, 4H), 3.29 (s, 3H), 2.28-2.85 (m, 2H).

(2) Ph (3-F) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

Mixture of Ph (3-F) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (OMe) (828 mg; 2.33 mmol; see step (1) above) and tetrabromide carbon (525 mg; 2.33 mmol) in 2-propanol (20 ml) was refluxed for 8 hours and then stirred at room temperature overnight. The mixture was concentrated in vacuo and the residue was partitioned between H ₂ O (70 ml) and EtOAc (50 ml). The aqueous layer was extracted with EtOAc (2 × 25 ml). Then, the combined organic extracts were washed with brine (35 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl ₃ / EtOH (15: 1) afforded the title compound (520 mg; 74%) as a crushable white foam.

Mp 73-81 ° C.

R _f = 0.43 (CHCl ₃ / EtOH (15: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.59 (d, J = 8 Hz, 2H), 7.32-7.37 (m, 2H), 7.05-7.14 (m, 2H), 6.87-6.92 ( m, 1H), 6.90 and 6.86 (t, J _HF = 73 Hz, 1H), 5.13-5.18 and 4.75-4.85 (m, 2H), 4.15-4.45 (m, 4H), 3.81 (s, 3H), 2.10 -2.75 (m, 2H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 172.0, 171.4, 153.9.

APCI-MS: (M + 1) = 481 m / z.

Example 27

Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 1,3-Dibromo-5-benzyloxybenzene

Sodium hydride (9.9 g; 0.414 mol, 95%, anhydrous) was added portionwise to a stirred solution of benzyl alcohol (41.0 g; 0.394 mol) in THF (1.0 L) at room temperature under nitrogen and stirred over 1 h. To this solution was added dropwise 1,3-dibromo-5-fluorobenzene (100.0 g; 0.394 mol). After stirring overnight, the mixture was partitioned between H ₂ O (600 ml) and EtOAc (4 × 600 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the subtitled compound (101.3 g; 75%) as a yellow oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.30-7.48 (m, 5H), 7.18 (s, 1H), 7.06 (s, 2H), 4.99 (s, 2H).

(2) 3,5-Dibromophenol

Aluminum chloride (11.7 g; 87.6 mmol) was added portionwise to a solution of 1,3-dibromo-5-benzyloxybenzene (10.0 g; 29.2 mmol; see step (1) above) and N, N-dimethylaniline (35.4 g; 292 mmol) in CH ₂ Cl ₂ (100 ml) at room temperature under a nitrogen atmosphere. After 30 min, the mixture was distributed between 1 N. HCl (300 ml) and EtOAc (5 × 150 ml). The combined organic extracts were washed with saturated NaHCO 3 (150 ml) and then with brine (150 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (9: 1) afforded the subtitle compound (6.1 g; 82%) as a white solid.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.21 (s, 1H), 6.97 (s, 2H), 5.88 (bs, 1 H).

(3) 1,3-Dibromo-5-monofluoromethoxybenzene

To a calibrated, hermetically sealed, pressurized 350 ml round-bottom flask containing a suspension of 3,5-dibromophenol (10.0 g; 39.7 mmol; see step (2) above) and Cs ₂ CO ₃ (20.7 g ; 63.5 mmol) in DMF (150 ml), chlorofluoromethane was added at -78 ° C by bubbling for 5 min through the membrane. The membrane was replaced with a Teflon stopper, the flask was then sealed and allowed to warm to room temperature, then the flask was weighed and it was determined that it contained 9.0 g (131 mmol) of chlorofluoromethane. This solution was heated in an oil bath set at 70 ° C. overnight. The flask was cooled to room temperature, carefully depressurized and the contents diluted with water (100 ml). Then the aqueous layer was extracted with Et ₂ O (3 × 100 ml), the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the subtitle compound (7.9 g; 71%) as a white solid.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.40 (s, 1H), 7.18 (s, 2H), 5.67 (d, J _HF = 53 Hz, 2H).

(4) 1-Bromo-3-monofluoromethoxy-5-vinylbenzene

Tri (butyl) vinyltin (10.0 g; 31.4 mmol) was added dropwise to a solution of 1,3-dibromo-5-monofluoromethoxybenzene (8.5 g; 29.9 mmol; see step (3) above), tetrakis (triphenylphosphine) palladium (0) (690 mg; 0.599 mmol) and 2,6-di-tert-butyl-4-methylphenol (at the tip of a spatula) in toluene (100 ml) in a nitrogen atmosphere. The mixture was stirred at 70 ° C. for 8 hours. The mixture was cooled to 0 ° C. and 1 N was added. NaOH (70 ml). After 1 h, the mixture was then extracted with CH ₂ Cl ₂ (3 × 300 ml), the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the subtitle compound (4.3 g; 57%) as a colorless oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.30 (s, 1H), 7.16 (s, 1H), 7.01 (s, 1H), 6.60 (dd. J = 6 Hz, J = 11 Hz, 1H), 5.74 (d, J = 16 Hz, 1H), 5.67 (d, J _HF = 53 Hz, 2H), 5.32 (d, J = 8 Hz, 1H).

(5) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) CH ₂ OH

2-Methyl-2-propanol (100 ml), H ₂ O (100 ml) and AD-mix-β (27.5 g) were combined and cooled to 0 ° C. 1-Bromo-3-monofluoromethoxy-5-vinylbenzene (4.3 g; 17.3 mmol; see step (4) above) was added immediately and this heterogeneous mixture was vigorously stirred at 0 ° C until confirmation by TLC of the absence of starting material . The reaction was quenched at 0 ° C by the addition of saturated sodium sulfite (200 ml), then warmed to room temperature and stirred for 60 minutes. The reaction mixture was extracted with EtOAc (3 × 150 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (4.9 g; 100%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.30 (s, 1H), 7.15 (s, 1H), 7.11 (s, 1H), 5.70 (d, J _HF = 53 Hz, 2H), 4.62-4.70 ( m, 1H), 3.52-3.70 (m, 2H).

HPLC analysis: 92.1%; 96.9% e.e., ChiralPak AD column (Hex / EtOH mobile phase (95: 5)).

(6) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) CH ₂ OTBS

To a solution of Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) CH ₂ OH (4.9 g; 18.6 mmol; see step (5) above), 4- (dimethylamino) pyridine (453 mg; 3.71 mmol) and DIPEA (8.9 g; 93.0 mmol) in anhydrous CH ₂ Cl ₂ (200 ml) was added dropwise a 1.0 M solution of tert-butyldimethylsilyl chloride in CH ₂ Cl ₂ ( 22.3 ml; 22.3 mmol). The reaction mixture was stirred for 10 hours at room temperature. DIPEA (8.9 g; 93.0 mmol) and 2-methoxyethoxymethyl chloride (13.9 g; 111 mmol) were added dropwise to the mixture. After 16 hours, an additional amount of 2-methoxyethoxymethyl chloride (2.2 g) was added and the reaction mixture was stirred overnight. The mixture was diluted with H ₂ O (100 ml) and the layers were separated. Then the aqueous layer was extracted with CH ₂ Cl ₂ (3 × 200 ml), the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (5: 1) afforded the subtitled compound (4.8 g; 55%) as a colorless oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.29 (s, 1H), 7.22 (s, 1H), 7.05 (s, 1H), 5.74 (d, J _HF = 53 Hz, 2H), 4.84 (d, J = 7 Hz, 1H), 4.70-4.74 (m, 2H), 3.50-3.91 (m, 6H), 3.42 (s, 3H), 0.90 (s, 9H), 0.05 (s, 3H), 0.01 ( s, 3H).

(7) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) CH ₂ OH

To a solution of Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) CH ₂ OTBS (4.7 g; 10.1 mmol; see step (6) above) in THF (100 ml) a 1.0 M solution of tetrabutylammonium fluoride in THF (13.1 ml; 13.1 mmol) was added at room temperature and the mixture was stirred for 1 h. The mixture was then partitioned between H ₂ O (100 ml) and EtOAc (3 × 100 ml), combined the organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (3.3 g; 92%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.22 (s, 1H), 7.14 (s, 1H), 7.03 (s, 1H), 5.71 (d, J _HF = 53 Hz, 2H), 4.80-4.82 ( m, 1H), 4.58-4.66 (m, 2H), 3.71-3.77 (m, 1H), 3.39-3.65 (m, 5H), 3.27 (s, 3H).

(8) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) C (O) OH

A solution of Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) CH ₂ OH (2.1 g; 6.0 mmol; see step (7) above) in acetone (40 ml) was added to an aqueous 5% solution of NaHCO ₃ (15 ml). This heterogeneous mixture mixed with a magnetic stir bar was cooled to 0 ° C, potassium bromide (70 mg; 0.60 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy free radical (976 mg; 5) were added to it. , 8 mmol). Then, sodium hypochlorite (5.25%; 15 ml) was added dropwise over a period of 10 minutes with vigorous stirring of the mixture and maintaining the temperature at 0 ° C. After 1 h, additional sodium hypochlorite (10 ml) and NaHCO ₃ solution (20 ml) were added and stirring was continued at 0 ° C for an additional 4 h. Acetone was removed on a rotary evaporator. The aqueous layer was diluted with 10% NaHCO ₃ solution (30 ml) and washed with Et ₂ O (3 × 20 ml). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (3 × 40 ml). Then, the combined EtOAc extracts were washed with H ₂ O (3 × 50 ml) and brine (50 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (1.7 g; 78%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.38 (s, 1H), 7.25 (s, 1H), 7.18 (s, 1H), 5.76 (d, J _HF = 53 Hz, 2H), 5.21 (s, 1H), 4.83 (d, J = 7 Hz, 1H), 4.75 (d, J = 7 Hz, 1H), 3.62-3.78 (m, 2H), 3.48-3.52 (m, 2H), 3.32 (s, 3H )

(9) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) C (O) OH (1.0 g; 2.72 mmol; see step (8) above) in DMF (20 ml) in a nitrogen atmosphere at 0 ° C, HAze-Pab (Teoc) · HCl (1.6 g; 3.5 mmol), RuBOP (1.6 g; 3.0 mmol) and DIPEA (880 mg; 6, 81 mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue was chromatographed twice on silica gel, eluting initially with CHCl ₃ / EtOH (15: 1) and a second time with EtOAc / EtOH (20: 1), which afforded the subtitle compound (1.2 g ; 62%) as crushable white foam.

¹ H NMR (300 MHz; CD ₃ OD, complex mixture of rotamers) δ 7.80-7.84 (m, 2H), 7.40-7.46 (m, 2H), 7.13-7.32 (m, 3H), 5.84-5.87 (m, 1H ), 5.67-5.69 (m, 1H), 5.25 and 5.07 (s, 1H), 5.18-5.23 and 4.80-4.88 (m, 1H), 3.97-4.79 (m, 8H), 3.60-3.71 (m, 2H) 3.40-3.53 (m, 2H), 3.32 (s, 3H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

(10) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc)

Mixture of Ph (3-Br) (5-OCH ₂ F) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc) (347 mg; 0.478 mmol; see step (9) above) and carbon tetrabromide (159 mg; 0.478 mmol) in 2-propanol (10 ml) was refluxed for 1.5 hours. The mixture was concentrated in vacuo, then partitioned with H ₂ O (20 ml) and EtOAc (3 × 30 ml) . The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel, eluting with CHCl ₃ / EtOH (15: 1), provided the subtitle compound (59 mg; 19%) as a crushable white foam.

Mp 81-87 ° C.

R _f = 0.58 (CHCl ₃ / EtOH (9: 1)).

¹ H NMR (300 MHz; CD ₃ OD, complex mixture of rotamers) δ 7.84 (d, J = 8 Hz, 2H), 7.40-7.48 (m, 2H), 7.18-7.30 (m, 3H), 5.80 (d, J _HF = 53 Hz, 2H), 5.21 and 5.15 (s, 1H), 5.18-5.24 and 4.80-4.88 (m. 1H), 3.98-4.54 (m, 6H), 2.10-2.70 (m, 2H), 1.05 -1.11 (m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 637 m / z.

(11) Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Br) (5-OCH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.073 g; 0.11 mmol; see step (10) above) was dissolved in 5 ml of TFA and allowed to interact for 90 minutes while cooling in an ice bath. TFA was evaporated and the residue was purified by preparative HPLC using CH ₃ CN / 0.1 M NH ₄ OAc (30:70). Suitable fractions were evaporated and freeze-dried from water / acetonitrile to give 49 mg (77%) of the title compound as its acetate salt.

¹ H-NMR (300 MHz; CD ₃ OD; rotamers) δ 7.8-7.7 (m, 2H), 7.54 (m, 2H), 7.37 (s, 1H, main rotamer), 7.33 (s, 1H, minor rotamer) , 7.25-7.1 (m, 2H), 5.75 (d, 2H), 5.22 (m, 1H, minor rotamer), 5.18 (s, 1H, primary rotamer), 5.11 (s, 1H, minor rotamer), 4.80 (m , 1H, main rotamer), 4.6-4.4 (m, 2H), 4.37 (m, 1H, main rotamer), 4.16 (m, 1H, main rotamer), 4.1-3.9 (m, 2H, two signals from the minor rotamer) 2.70 (m, 1H, minor rotamer), 2.52 (m, 1H, primary rotamer), 2.30 (m, 1H, primary rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H).

ESI-MS +: (M + 1) = 493/495 (m / z).

Example 28

Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

(1) 1,3-Dibromo-5-difluoromethoxybenzene

To a calibrated, hermetically sealed, pressurized 350 ml round bottom flask containing a solution of 3,5-dibromophenol (10.0 g; 39.7 mmol; see example 27 (2) above) in 2-propanol (100 ml) and 30% KOH (80 ml), at -78 ° C, chlorodifluoromethane was added by bubbling for 15 min through the membrane. The membrane was replaced with a Teflon plug, then the flask was hermetically sealed and allowed to warm to room temperature, then the flask was weighed and it was determined that it contained 12.0 g (138 mmol) of chlorodifluoromethane. This solution was refluxed overnight in an oil bath set to 80 ° C. The flask was cooled to room temperature, carefully depressurized and the contents diluted with H ₂ O (200 ml). The aqueous layer was extracted with CHCl ₃ (2 × 150 ml), then the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The residue was purified by distillation using a spherical expansion tube (Kugelrohr) at 80 ° C and 26.7 Pa (0.2 mm Hg), which allowed to obtain the compound indicated in the subtitle (9.6 g; 80%) as clear liquid.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.55 (s, 1H), 7.26 (s, 2H), 6.52 (t, J _HF = 68 Hz, 1H).

(2) 1-Bromo-3-difluoromethoxy-5-vinylbenzene

Tri (butyl) vinyltin (10.5 g; 33.1 mmol) was added dropwise to a solution of 1,3-dibromo-5-difluoromethoxybenzene (9.1 g; 30.1 mmol; see step (1) above), tetrakis (triphenylphosphine) palladium (0) (700 mg; 0.60 mmol) and 2,6-di-tert-butyl-4-methylphenol (at the tip of a spatula) in toluene (125 ml) in a nitrogen atmosphere. The mixture was stirred at 50 ° C. overnight. The mixture was cooled to 0 ° C. and 1N was added. NaOH (70 ml). After 1 h, the mixture was then extracted with CH ₂ Cl ₂ (3 × 300 ml), the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the subtitle compound (5.1 g; 68%) as a colorless oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.53 (s, 1H), 7.18 (s, 1H), 7.08 (s, 1H), 6.60 (dd, J = 6 Hz, J = 11 Hz, 1H), 6.57 (t, J _HF = 68 Hz, 1H), 5.77 (d, J = 11 Hz, 1H), 5.36 (d, J = 8 Hz, 1H).

(3) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OH

2-Methyl-2-propanol (150 ml), H ₂ O (150 ml) and AD-mix-β (27.8 g) were combined and cooled to 0 ° C. 1-Bromo-3-difluoromethoxy-5-vinylbenzene (4.6 g; 18.6 mmol; see step (2) above) was added immediately and this heterogeneous suspension was vigorously stirred at 0 ° C until confirmation by TLC of the absence of starting material , then the solution was warmed to room temperature and stirred overnight. The reaction was quenched at 0 ° C by adding saturated sodium sulfite (300 ml) and then warmed to room temperature and stirred for 60 minutes. The reaction mixture was extracted with EtOAc (3 × 200 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (5.0 g; 95%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.43 (s, 1H), 7.23 (s, 1H), 7.16 (s, 1H), 6.86 (t, J _HF = 75 Hz, 1H), 4.64-4.67 ( m, 1H), 3.54-3.59 (m, 2H).

HPLC analysis: 88.6%; 96.3% e.e., ChiralPak AD column (Hex / EtOH mobile phase (95: 5)).

(4) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OTBS

To a solution of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) CH ₂ OH (4.9 g; 17.3 mmol; see step (3) above), 4- (dimethylamino) pyridine (420 mg; 3.5 mmol) and DIPEA (11.2 g; 86.3 mmol) in anhydrous CH ₂ Cl ₂ (250 ml) was added dropwise a 1.0 M solution of tert-butyldimethylsilyl chloride in CH ₂ Cl ₂ (20 7 ml; 20.7 mmol). The reaction mixture was stirred overnight at room temperature. DIPEA (11.2 g; 86.3 mmol) and 2-methoxyethoxymethyl chloride (12.9 g; 104 mmol) were added dropwise to the mixture. After 3 days. additional 2-methoxyethoxymethyl chloride (3.3 g) was added and the reaction mixture was stirred overnight. The mixture was diluted with water (250 ml) and the layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (2 × 250 ml), then the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with a Hex / EtOAc mixture (4: 1) afforded the subtitled compound (4.3 g; 51%) as a colorless oil.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.40 (s, 1H), 7.25 (s, 1H), 7.08 (s, 1H), 6.58 (t, J _HF = 75 Hz, 1H), 4.84 (d, J = 7 Hz, 1H), 4.70-4.74 (m, 2H), 3.50-3.91 (m, 6H), 3.42 (s, 3H), 0.90 (s, 9H), 0.12 (s, 3H), 0.05 ( s, 3H).

(5) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OH

To a solution of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OTBS (3.3 g; 6.9 mmol; see step (4) above) in THF (60 ml) was added A 1.0 M solution of tetrabutylammonium fluoride in THF (9.0 ml; 9.0 mmol) at room temperature. The reaction mixture was stirred for 45 minutes, then the mixture was partitioned between water (150 ml) and EtOAc (2 × 120 ml). The combined organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitle compound (2.5 g; 98%) as a yellow oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.35 (s, 1H), 7.21 (s, 1H), 7.08 (s, 1H), 6.83 (t, J _HF = 73 Hz, 1H), 4.73 (d, J = 7 Hz, 1H), 4.59-4.68 (m, 2H), 3.40-3.80 (m, 6H), 3.26 (s, 3H).

(6) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH

A solution of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) CH ₂ OH (3.0 g; 8.1 mmol; see step (5) above) in acetone (60 ml) was added to aqueous 5% solution of NaHCO ₃ (25 ml). This heterogeneous mixture stirred with a magnetic stir bar was cooled to 0 ° C, then potassium bromide (100 mg; 0.81 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy free radical (1.3 g; 8.5 mmol). Then, sodium hypochlorite (5.25%; 19 ml) was added dropwise over a period of 10 minutes with vigorous stirring of the mixture and maintaining the temperature at 0 ° C. After 1 h, additional sodium hypochlorite (17 ml) and NaHCO ₃ solution (34 ml) were added, stirring was continued at 0 ° C for an additional 4 h. Acetone was removed on a rotary evaporator. The aqueous layer was diluted with 10% NaHCO ₃ solution (30 ml) and washed with Et ₂ O (3 × 20 ml). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (3 × 40 ml). The combined EtOAc layers were washed with H ₂ O (3 × 50 ml) and brine (50 ml) and then dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, which afforded the subtitled compound (2.1 g; 66%) as a colorless oil, which was used without further purification.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.51 (s, 1H), 7.32 (s, 1H), 7.24 (s, 1H), 6.88 (t, J _HF = 73 Hz, 1H), 5.21 (s, 1H), 4.84 (d, J = 7 Hz, 1H), 4.76 (d, J = 7 Hz, 1H), 3.62-3.80 (m, 2H), 3.48-3.52 (m, 2H), 3.32 (s, 3H )

(7) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc)

To a solution of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH (1.0 g; 2.62 mmol; see step (6) above) in DMF (50 ml ) in a nitrogen atmosphere at 0 ° C, HAze-Pab (Teoc) · HCl (1.5 g; 3.38 mmol), RuBOP (1.5 g; 2.9 mmol) and DIPEA (840 mg; 6.50) were added mmol). The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue was chromatographed on silica gel eluting with CHCl ₃ / EtOH (15: 1), which afforded the subtitled compound (1.1 g; 59%) as a crushable white foam.

¹ H NMR (300 MHz; CD ₃ OD, complex mixture of rotamers) δ 7.79-7.83 (m, 2H), 7.26-7.52 (m, 5H), 6.94 and 6.91 (t, J _HF = 73 Hz, 1H), 5.27 and 5.07 (s, 1H), 5.20-5.23 and 4.80-4.88 (m, 1H), 4.01-4.79 (m, 8H), 3.60-3.71 (m, 2H), 3.40-3.53 (m, 2H), 3.32 ( s, 3H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

(8) Ph (3-Br) (5-OCHF ₂₎ - (R) CH (OH) C (O) -Aze-Pab (Teoc)

Mixture of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (Teoc) (369 mg; 0.496 mmol; see step (7) above) and carbon tetrabromide ( 165 mg; 0.496 mmol) in 2-propanol (10 ml) was refluxed for 12 hours. The mixture was concentrated in vacuo, then partitioned with H ₂ O (15 ml) and EtOAc (5 × 20 ml). The combined organic phases were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel, eluting with CHCl ₃ / EtOH (15: 1), provided the subtitle compound (134 mg; 41%) as a crushable white foam.

Mp. 92-98 ° C.

R _f = 0.37 (CHCl ₃ / EtOH (9: 1)).

¹ H NMR (300 MHz; CD ₃ OD, complex mixture of rotamers) δ 7.80-7.86 (m, 2H), 7.40-7.48 (m, 2H), 7.10-7.33 (m, 3H), 6.92 and 6.88 (t, J _HF = 73 Hz, 1H), 5.18 and 5.11 (s, 1H), 5.18-5.24 and 4.76-4.80 (m, 1H), 3.98-4.54 (m, 6H), 2.10-2.70 (m, 2H), 1.05- 1.11 (m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 655 m / z.

(9) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab × TFA

Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (0.081 g; 0.124 mmol; see step (8) above) was dissolved in 5 ml of TFA and allowed to interact for 80 minutes while cooling in an ice bath. TFA was evaporated and the residue was purified by preparative HPLC using CH ₃ CN / 0.1 M NH ₄ OAc (30:70). The appropriate fractions were evaporated and freeze-dried from water / acetonitrile to give 59 mg (83%) of the title compound as its acetate salt.

¹ H-NMR (300 MHz; CD ₃ OD; rotamers): δ 7.8-7.7 (m, 2H), 7.6-7.4 (m, 3H), 7.3-7.2 (m, 2H), 6.89 (t, 1H, basic rotamer), 6.87 (t, 1H, minor rotamer), 5.23 (m, 1H, minor rotamer), 5.21 (s, 1H, primary rotamer), 5.13 (s, 1H, minor rotamer), 4.80 (m, 1H, main rotamer), 4.6-4.4 (m, 2H), 4.38 (m, 1H, main rotamer), 4.20 (m, 1H, main rotamer), 4.1-3.9 (m, 2H, two signals from the minor rotamer), 2.70 (m , 1H, minor rotamer), 2.54 (m, 1H, main rotamer), 2.29 (m, 1H, main rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H).

¹³ C-NMR (75 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 172.0, 171.7, 167.0.

MS (m / z) 511/513 (M + 1) ⁺ .

Example 29

Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

(1) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (OMe)

To a solution of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) OH (957 mg; 2.48 mmol; see example 28 (6) above) in DMF (30 ml) in a nitrogen atmosphere at 0 ° C, HAze-Pab (OMe) · 2HCl (1.1 g; 3.2 mmol), RuBOP (1.4 g; 2.7 mmol) and DIPEA (804 mg; 6.2 mmol) were added ) The reaction mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo, the residue was subjected to silica gel chromatography twice, eluting initially with CHCl ₃ / EtOH (9: 1) and a second time with EtOAc / EtOH (15: 1), which afforded the subtitle compound (1.1 g; 72%) as crushable white foam.

¹ H NMR (300 MHz; CD ₃ OD, complex mixture of rotamers) δ 7.59-7.65 (m, 2H), 7.20-7.55 (m, 5H), 6.95 and 6.91 (t, J _HF = 73 Hz, 1H), 5.27 and 5.07 (s, 1H), 5.18-5.23 and 4.75-4.84 (m, 1H), 3.87-4.89 (m, 6H), 3.84 (s, 3H), 3.60-3.71 (m, 2H), 3.40-3.53 ( m, 2H), 3.32 (s, 3H), 2.10-2.75 (m, 2H).

(2) Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe)

Mixture of Ph (3-Br) (5-OCHF ₂ ) - (R) CH (OMEM) C (O) -Aze-Pab (OMe) (1.1 g; 1.8 mmol; see step (1) above) and carbon tetrabromide (583 mg; 1.8 mmol) in 2-propanol (30 ml) was refluxed for 2.5 days. During this time, to complete the reaction, an additional amount of carbon tetrabromide was added (5 portions of 50 mg at intervals for an additional 0.90 mmol). The mixture was concentrated in vacuo, then distributed with H ₂ O (50 ml) and EtOAc (5 × 25 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl ₃ / EtOH (15: 1) afforded the title compound (460 mg; 50%) as a crushable white foam.

Mp 71-75 ° C.

R _f = 0.63 (CHCl ₃ / EtOH (9: 1)).

¹ H NMR (300 MHz; CD ₃ OD; complex mixture of rotamers) δ 7.59 (d, J = 8 Hz, 2H), 7.20-7.54 (m, 5H), 6.90 and 6.87 (t, J _HF = 73 Hz, 1H ), 5.18 and 5.11 (s, 1H), 4.76-4.80 (m, 1H), 3.98-4.54 (m, 4H), 3.82 (s, 3H), 2.10-2.70 (m, 2H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.5, 172.1, 171.6, 154.1.

APCI-MS: (M + 1) = 542 m / z.

Example 30

Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH)

(1) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Z)

Boc-Aze-Pab (Z) (see international patent application WO 97/02284, 92 mg; 0.197 mmol) was dissolved in 10 ml of EtOAc saturated with HCl (g) and allowed to react for 10 minutes. The solvent was evaporated, the residue was mixed with Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) OH (50 mg; 0.188 mmol; see example 17 (5) above), RuBOP (109 mg; 0.209 mmol) and finally with diisopropylethylamine (96 mg; 0.75 mmol) in 2 ml of DMF. The mixture was stirred for 2 hours, then poured into 50 ml of water and extracted with EtOAc three times. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel eluting with EtOAc / MeOH (9: 1). Yield 100 mg (87%).

¹ H NMR (300 MHz; CD ₃ OD, mixture of rotamers) δ 7.85-7.75 (m, 2H), 7.45-7.25 (m, 7H), 7.11 (m, 1H, main rotamer), 7.08 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.13 (bt, 1H), 5.25-5.05 (m, 3H), 4.77 (m, 1H, partially invisible behind the signal from CD ₃ OH), 4.5-3.9 (m, 7H), 2.64 (m, 1H, minor rotamer), 2.47 (m, 1H, primary rotamer), 2.25 (m, 1H, primary rotamer), 2.13 (m, 1H, minor rotamer).

(2) Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH)

Hydroxylamine hydrochloride (65 mg; 0.94 mmol) and triethylamine (0.319 g; 3.16 mmol) were mixed in 8 ml of THF and sonicated for 1 h at 40 ° C. Ph (3-Cl) (5-OCH ₂ CHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Z) (96 mg; 0.156 mmol; was added; see step (1) above) with additional 8 ml THF. The mixture was stirred at 40 ° C. for 4.5 days. The solvent was evaporated and the crude product was purified by preparative RPLC in a mixture of CH ₃ CN / 0.1 M NH ₄ OAC (40:60). Yield 30 mg (38%). Purity 99%.

¹ H NMR (300 MHz; CD ₃ OD, mixture of rotamers) δ 7.6-7.55 (m, 2H), 7.35-7.3 (m, 2H), 7.12 (m, 1H, main rotamer), 7.09 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.15 (triplet of multiplets, 1H), 5.15 (m, 1H, minor rotamer), 5.13 (s, 1H, main rotamer), 5.08 (s, 1H, minor rotamer), 4.77 (m, 1H, main rotamer), 4.5-4.2 (m, 5H), 4.08 (m, 1H, main rotamer), 3.97 (m, 1H, minor rotamer), 2.66 (m, 1H, minor rotamer), 2.50 (m, 1H, main rotamer), 2.27 (m, 1H, main rotamer), 2.14 (m, 1H, minor rotamer),

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 172.8, 172.2, 171.4, 159.1, 158.9, 154.2.

APCI-MS: (M + 1) = 497/499 m / z.

Example 31

Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OH)

(1) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Z)

Boc-Aze-Pab (Z) (130 mg; 0.279 mmol) was dissolved in 15 ml EtOAc saturated with HCl (g) and allowed to react for 10 minutes. The solvent was evaporated and the residue was mixed with Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) OH (63 mg; 0.188 mmol; see Example 21 (5) above) in 3 ml DMF, RuBOP (147 mg; 0.279 mmol) and finally with diisopropylethylamine (134 mg; 1.03 mmol). The mixture was stirred for 130 minutes and then poured into 75 ml of water and extracted with EtOAc three times. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc / MeOH (95: 5). Yield 119 mg (79%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 8.06 (bt, 1H), 7.67 (d, 2H), 7.45-7.25 (m, 5H), 7.18 (d, 2H), 6.89 (m, 1H), 6.84 ( m, 1H), 6.76 (m, 1H), 5.16 (s, 2H), 4.84 (s, 1H), 4.79 (m, 1H), 4.66 (doublet of multiplets, 2H), 4.4-4.3 (m, 2H), 4.10 (doublet of multiplets, 2H), 4.02 (m, 1H), 3.67 (m, 1H), 2.46 (m, 1H), 2.28 (m, 1H).

(2) Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (OH)

Hydroxylamine hydrochloride (80 mg; 1.16 mmol) and triethylamine (0.392 g; 3.87 mmol) were mixed in 9 ml of THF and sonicated for 1 h at 40 ° C. Ph (3-Cl) (5-OCH ₂ CH ₂ F) - (R) CH (OH) C (O) -Aze-Pab (Z) (96 mg; 0.156 mmol; was added; see step (1) above) s an additional 9 ml of THF. The mixture was stirred at 40 ° C. for 48 hours and 3 days at room temperature. The solvent was evaporated and the crude product was purified by preparative RPLC in a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (30:70). Yield 72 mg (78%). Purity 100%.

¹ H NMR (400 MHz; CD ₃ OD, mixture of rotamers) δ 7.6-7.55 (m, 2H), 7.35-7.25 (m, 4H), 7.07 (m, 1H, main rotamer), 7.04 (m, 1H, minor rotamer), 7.0-6.9 (m, 2H), 5.12 (m, 1H, minor rotamer), 5.08 (s, 1H, minor rotamer), 5.04 (s, 1H), 4.78 (m, 1H, main rotamer), 4.68 (doublet of multiplets, 2H), 4.5-4.25 (m, 3H), 4.20 (doublet of multiplets, 2H), 4.06 (m, 1H, main rotamer), 3.97 (m, 1H, minor rotamer), 2.65 (m, 1H, minor rotamer), 2.48 (m, 1H, main rotamer), 2.27 (m, 1H, main rotamer), 2.14 (m, 1H, minor rotamer).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 172.3, 171.5, 159.8, 154.3.

APCI-MS: (M + 1) = 479/481 m / z.

Example 32

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Pro-Pab

(1) Boc-Pro-Pab (Teoc)

Boc-Pro-Pab (Z) (see WO 97/02284; 15.0 g; 0.0321 mol) was dissolved in 150 ml of ethanol and 200 mg of 10% Pd / C (50%) was added. humidity). The mixture was stirred and hydrogenated at atmospheric pressure for 2 hours, filtered through Hyflo and concentrated. The product was used without further purification. 10 g of this product (0.029 mol) was dissolved in 300 ml of THF. Theos-n-nitrophenyl carbonate (10 g; 0.035 mol) was added. Over a period of 3 minutes, a solution of potassium carbonate (5.2 g; 0.038 mol) in 50 ml of water was added, the resulting solution was stirred for 3 days, concentrated, and the residue was extracted three times with EtOAc. The combined organic layer was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel using methylene chloride / acetone (4: 1). Yield 9.8 g (69%).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Pro-Pab (Teoc)

Boc-Pro-Pab (Teoc) (107 mg; 0.218 mmol; see step (1) above) was dissolved in 10 ml of HCl saturated EtOAc (g) and allowed to react for 10 minutes. The solvent was evaporated, the residue was mixed with Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (50 mg; 0.198 mmol; see example 1 (8) above) in 3 ml of DMF , RuBOP (115 mg; 0.218 mmol) and finally with diisopropylethylamine (104 mg; 0.80 mmol). The mixture was stirred for 2 hours, then poured into 75 ml of water and extracted with EtOAc three times. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc / MeOH (95: 5). Yield 89 mg (72%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.54 (bt, 1H), 7.47 (d, 2H), 7.12 (m, 1H), 7.08 (d, 2H), 7.02 (m, 1H), 6.95 (m, 1H), 6.50 (t, 1H), 5.21 (s, 1H), 4.42 (m, 1H), 4.35-4.15 (m, 3H), 3.59 (m, 1H), 2.94 (m, 1H), 2.1-1.7 (m, 4H), 1.06 (m, 2H), 0.04 (s, 9H).

(3) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Pro-Pab × TFA

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Pro-Pab (Teoc) (85 mg; 0.136 mmol; see step (2) above) was dissolved in 1 ml of methylene chloride and cooled in an ice bath. TFA (4 ml) was added and the reaction mixture was stirred for 90 minutes. TFA was evaporated and the residue was freeze dried from water and acetonitrile. Yield 72 mg (92%). Purity 94%.

¹ H NMR (400 MHz; CD ₃ OD, mixture of rotamers) δ 7.85-7.7 (m, 2H), 7.58 (d, 2H, main rotamer), 7.47 (d, 2H, minor rotamer), 7.35 (m, 1H, main rotamer), 7.27 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H), 6.88 (t, 1H), 5.38 (s, 1H, main rotamer), 5.22 (s, 1H, minor rotamer), 4.58 (d, 1H), 4.5-4.2 (m, 2H), 3.8-3.5 (m, 1H), 3.35 (m, 1H), 2.2-1.8 (m, 4H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons) δ 173.6, 171.1, 167.0.

APCI-MS: (M + 1) = 481/483 m / z.

Example 33

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Pro-Pab (OMe)

(1) 4-Azidomethyl-N-methoxybenzamidine

4-Azidomethylbenznitrile (17.3 g; 0.109 mol; Nishiyama et al .; Chem. Lett. (1982) 1477) was dissolved in 500 ml of toluene and 200 ml of absolute ethanol. The solution was cooled to -10 ° C and HCl (g) was bubbled through it to saturation. The mixture was kept in the refrigerator for 2 days, during which most of the solvents evaporated. Diethyl ether was added and decantation was carried out. The product was redissolved in a solution of O-methylhydroxylamine (10.5 g; 0.125 mol) and triethylamine (56 ml) in 200 ml of methanol. The mixture was allowed to stand for 3 days, after which the methanol was evaporated with EtOAc. The organic phase was washed with water, diluted with HOAc and aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and again diluted with EtOAc to a total volume of 500 ml. A 25 ml sample was evaporated to dryness. The residue was 932 mg. The total yield of 18.6 g (83%).

(2) 4-Aminomethyl-N-methoxybenzamidine

To a solution of 4-azidomethyl-N-methoxybenzamidine (11.3 g; 0.055 mol; see step (1) above) in 200 ml of ethanol was added 200 mg of PtO ₂ . The mixture was hydrogenated with constant bubbling with hydrogen for 4 hours and then filtered through Celite® and evaporated. Yield 7.34 g (74%).

(3) Boc-Pro-Pab (OMe)

To a suspension of Boc-Pro-OH (9.7 g; 0.045 mol), 4-aminomethyl-N-methoxybenzamidine (7.33 g; 0.041 mol; see step (2) above) and dimethylaminopyridine (7.8 g; 0.064 mol ) in 300 ml of acetonitrile was added EDC, base (11.7 ml; 0.068 mol). The mixture was stirred for 18 hours, concentrated and partitioned between water and EtOAc. The organic layer was washed with water, aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc. Yield 9.73 g (63%).

(4) H-Pro-Pab (OMe) × 2HCl

Boc-Pro-Pab (OMe) (9.7 g; 0.026 mol; see step (3) above) was dissolved in 250 ml EtOAc. This ice-cooled solution was saturated with HCl (g) by sparging for 5 minutes. The product immediately precipitated and 125 ml of absolute ethanol was added. The mixture was sonicated until most of the material solidified. Diethyl ether (200 ml) was added and the suspension was filtered. Several lumps that did not solidify were again treated with absolute ethanol and diethyl ether. The solid was dried. Yield 7.57 g (86%).

¹ H NMR (400 MHz; CD ₃ OD) δ 7.74 (d, 2H), 7.58 (d, 2H), 4.55 (s, 2H), 4.38 (m, 1H), 3.98 (s, 3H), 3.45-3.3 (m, 2H), 2.50 (m, 1H), 2.15-2.0 (m, 3H).

(5) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Pro-Pab (OMe)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (50 mg; 0.198 mmol; see Example 1 (8) above), H-Pro-Pab (OMe) ( 76 mg; 0.218 mmol; see step (4) above) and RuBOP (115 mg; 0.218 mmol) was dissolved in 2 ml of DMF. Diisopropylethylamine (104 mg; 0.80 mmol) was added and the mixture was stirred for 2.5 hours. The mixture was poured into 50 ml of water, extracted with EtOAc three times, the combined organic phase was washed with brine, dried (Na ₂ SO ₄ ) and evaporated. The residue was flash chromatographed on silica gel in EtOAc / MeOH (95: 5). Yield 37 mg (36%). Purity 98%.

¹ H NMR (400 MHz; CD ₃ OD, mixture of rotamers) δ 7.60 (d, 2H, main rotamer), 7.57 (d, 2H, minor rotamer), 7.4-7.1 (m, 5H), 6.89 (t, 1H, main rotamer), 6.87 (t, 1H, minor rotamer), 5.35 (s, 1H, main rotamer), 5.21 (s, 1H, minor rotamer), 4.72 (m, 1H, minor rotamer), 4.5-4.35 (m, 1H and 2H, main rotamer), 4.3-4.25 (m, 2H, minor rotamer), 3.814 (s, 3H, main rotamer), 3.807 (s, 3H, minor rotamer), 3.75-3.5 (m, 1H), 3.35 (m, 1H), 2.2-1.8 (m, 4H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 173.3, 173.2, 171.3, 171.0, 153.9, 152.4.

APCI-MS: (M + 1) = 511/513 m / z.

Example 34

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((2-amidino) -5-

pyridinyl)

(1) 6-cyanonicotinic acid

To a solution of nicotinic acid N-oxide (51 g; 0.37 mol) in 1.2 L of DMF was added NaCN (54 g; 1.1 mol) followed by triethylamine (255 ml; 1.83 mol) and TMSCl (185 ml). The reaction mixture was stirred at 110 ° C. for 10 hours, filtered and the filtrate was concentrated. The residue was dissolved in 100 ml of 2 N. HCl and was extracted with methylene chloride. The organic layers were combined, concentrated and recrystallized from water to obtain 12 g (22%) of the product.

(2) 5- (Hydroxymethyl) pyridin-2-carbonitrile

To a solution of 6-cyanonicotinic acid (12 g; 0.081 mol; see step (1) above) in THF at 0 ° C was added Et ₃ N (12.4 ml; 0.0892 mol) followed by ethyl chloroformate (8.53 ml ; 0.0892 mol). The reaction mixture was stirred for 15 minutes and NaBH ₄ (6.14 g; 0.162 mol) was added. The mixture was then stirred at RT overnight, quenched with water and extracted with methylene chloride. The organic layer was concentrated and purified by column chromatography to give 4 g (20%) of alcohol.

(3) 5- (Azidomethyl) pyridin-2-carbonitrile

5- (Hydroxymethyl) pyridin-2-carbonitrile (4 g; 0.03 mol; see step (2) above) was dissolved in 25 ml of methylene chloride and cooled in an ice bath. Mesyl chloride (2.32 ml; 0.0300 mol) was added dropwise and then triethylamine (4.6 ml; 0.033 mol). The reaction mixture was stirred and, after mixing, the crude mesylate was treated with NaN ₃ (7.35 g; 0.113 mol) in 20 ml of DMF. The reaction mixture was stirred at 40 ° C. for 2 hours, diluted with water and extracted with ethyl acetate. The organic layer was concentrated to give 3.95 g (83%) of crude azide.

(4) 5- (tert-Butoxycarbonylaminomethyl) pyridin-2-carbonitrile

To a solution of 5- (azidomethyl) pyridin-2-carbonitrile (3.95 g; 0.0248 mol; see step (3) above) in 30 ml THF and 10 ml water was added triphenylphosphine (7.8 g; 0.0298 mol ) and the resulting mixture was stirred for 24 hours. Then, triethylamine (3.8 ml; 0.027 mol) was added, followed by the addition of Boc anhydride (5.4 g; 0.025 mol) with stirring for 2 hours. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was concentrated and purified by column chromatography to give 2.1 g (36%) of the subtitle compound.

¹ H NMR (300 MHz; CDCl ₃ ) δ 8.6 (s, 1H), 8.0 (d, 1H), 8.9 (d, 1H), 4.1 (m, 2H), 1.4 (s, 9H).

(5) 5- (Aminomethyl) pyridin-2-carbonitrile × 2HCl

5- (tert-Butoxycarbonylaminomethyl) pyridin-2-carbonitrile (0.200 g; 0.86 mmol; see step (4) above) was dissolved in 10 ml of HCl saturated EtOAc (g) and stirred for 30 minutes. The solvent was evaporated, and 0.175 g (99%) of the subtitle compound was obtained as its dihydrochloride salt.

¹ H NMR (500 MHz; D ₂ O) δ 8.79 (s, 1H), 8.17 (d, 1H), 8.05 (d, 1H), 4.38 (s, 2H).

(6) Boc-Aze-NH-CH ₂ -5-Py (2-CN)

To a mixture of 5- (aminomethyl) pyridin-2-carbonitrile × 2HCl (0.175 g; 0.85 mmol; see step (5) above), Boc-Aze-OH (0.201 g; 1.00 mmol) and TBTU (0.321 g ; 1.00 mmol) in 5 ml of DMF was added dimethylaminopyridine (0.367 g; 3.00 mmol). The mixture was stirred overnight and then poured into water and extracted with EtOAc three times. The combined organic phase was washed with aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated. The crude product starting to crystallize was used as such in the next step. Yield 0.23 g (73%).

¹ H NMR (500 MHz; CDCl ₃ ) δ 8.66 (s, 1H), 8.2-7.8 (broad, 1H), 7.79 (d, 1H), 7.67 (d, 1H), 4.73 (m, 1H), 4.65- 4.5 (m, 2H), 3.94 (m, 1H), 3.81 (m, 1H), 2.6-2.35 (m, 2H), 1.8 (broadened, 1H), 1.45 (s, 9H).

(7) H-Aze-NH-CH ₂ -5-Py (2-CN) × 2HCl

Boc-Aze-NH-CH ₂ -5-Py (2-CN) (0.23 g; 0.73 mmol; see step (6) above) was dissolved in 10 ml of EtOAc saturated with HCl (g) and stirred in within 30 minutes The solvent was evaporated, and 0.21 g (100%) of the subtitle compound was obtained as its dihydrochloride salt.

¹ H NMR (500 MHz; D ₂ O) δ 8.64 (s, 1H), 8.0-7.9 (m, 2H), 5.19 (m, 1H), 4.65-4.55 (m, 2H), 4.20 (m, 1H) 4.03 (m, 1H); 2.88 (m, 1H); 2.64 (m, 1H).

(8) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Aze-NH-CH ₂ -5-Py (2-CN)

To the mixture H-Aze-NH-CH ₂ -5-Py (2-CN) × 2HCl (0.206 g; 0.713 mmol; see step (7) above), Ph (3-Cl) (5-OCHF ₂ ) - ( R) CH (OH) C (O) OH (0.180 g; 0.713 mmol; see Example 1 (8) above) and RuBOP (0.408 g; 0.784 mmol) in 5 ml of DMF was added dimethylaminopyridine (0.367 g; 3.00 mmol) . The mixture was stirred overnight, then poured into water and extracted with EtOAc three times. The combined organic phase was washed with aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc to give the pure product. Yield 0.197 g (61%).

¹ H NMR (500 MHz; CDCl ₃ ) δ 8.63 (m, 1H), 8.22 (bt, 1H), 7.78 (m, 1H), 7.67 (m, 1H), 7.21 (m, 1H), 7.16 (m, 1H), 7.04 (m, 1H), 6.56 (t, 1H), 4.97 (bd, 1H), 4.92 (m, 1H), 4.6-4.5 (m, 2H), 4.40 (bd, 1H), 4.18 (m , 1H), 3.80 (m, 1H), 2.69 (m, 1H), 2.46 (m, 1H), 1.92 (s, 1H).

APCI-MS: (M + 1) = 451/453 m / z.

(9) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Aze-NH-CH ₂ - ((2-amidino) -5-pyridinyl) × HOAc

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Aze-NH-CH ₂ -5-Py (2-CN) (0.200 g; 0.444 mmol; see step (8) above), ammonium acetate (1.00 g; 0.0130 mol) and N-acetylcysteine (2.0 g; 0.0122 mol) in 10 ml of methanol was heated at 50 ° C for 2 days. Preparative OFLC in a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (30:70) and repeated transmission of the corresponding fractions in a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (5: 95-40: 60) yielded 60 mg (26%) of the pure title compound as its acetate salt after freeze-drying from water and acetonitrile. Purity 100%.

¹ H NMR (500 MHz; D ₂ O, mixture of rotamers) δ 8.68 (s, 1H, main rotamer), 8.62 (s, 1H, minor rotamer), 8.05-7.9 (m, 2H), 7.33 (m, 1H, rotamer), 7.27 (m, 1H, rotamer), 7.22 (m, 1H, rotamer), 7.17 (m, 1H, rotamer), 7.01 (m, 1H, rotamer), 6.84 (t, 1H), 5.32 (s, 1H, main rotamer), 5.20 (m, 1H, minor rotamer), 5.13 (s, 1H, minor rotamer), 4.88 (m, 1H, main rotamer), 4.65-4.55 (m, 2H, main rotamer), 4.45- 4.35 (m, 1H, rotamer plus 1H, minor rotamer), 4.31 (d, 1H, minor rotamer), 4.2-4.05 (m, 1H plus 1H, rotamer), 2.80 (m, 1H, minor rotamer), 2.61 (m , 1H, main rotamer), 2.33 (m, 1H, main rotamer), 2.24 (m, 1H, minor rotamer), 1.93 (s, 3H).

¹³ C-NMR (100 MHz; D ₂ O) (carbonyl and / or amidine carbons, mixture of rotamers) δ 181.6, 173.3, 172.7, 172.6, 172.3, 162.6, 162.3.

APCI-MS: (M + 1) = 468/470 m / z.

Example 35

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((2-methoxyamidino) -5-pyridinyl)

(1) Boc-NH-CH ₂ - [(2- (amino (hydroxylimino) methyl)) - 5-pyridinyl]

5- (tert-Butoxycarbonylaminomethyl) pyridin-2-carbonitrile (1.00 g; 4.29 mmol; see example 34 (4) above) was dissolved in 10 ml of ethanol and hydroxylamine hydrochloride (0.894 g; 0.0129 mol) was added and triethylamine (1.30 g; 0.0129 mol). The mixture was stirred at room temperature for 6 days. The mixture was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 0.96 g (84%).

¹ H NMR (400 MHz; acetone-d ₆ ) δ 9.01 (bs, 1H), 8.50 (bs, 1H), 7.87 (m, 1H), 7.70 (m, 1H), 6.58 (broad, 1H), 5.70 ( broad, 2H), 4.31 (d, 2H), 1.41 (s, 1H).

(2) Boc-Aze-NH-CH ₂ - (2- (amidino) -5-pyridinyl) × HOAc

This reaction was carried out according to the method described in Judkins et al., Synth. Comm. (1998) 4351. Suspension Boc-NH-CH ₂ - [(2- (amino (hydroxylimino) methyl)) - 5-pyridinyl] (0.910 g; 3.42 mmol; see step (1) above), acetic anhydride ( 0.35 ml; 3.7 mmol) and 0.35 g of 10% Pd / C (50% humidity) in 100 ml of acetic acid were hydrogenated under a pressure of 490 kPa (5 atm) for 5 hours. The mixture was filtered. through Celite and concentrated. The residue was freeze dried from water and acetonitrile to give 0.97 g (92%) of the subtitled compound.

¹ H NMR (500 MHz; CD ₃ OD) δ 8.74 (s, 1H), 8.12 (d, 1H), 7.98 (d, 1H), 4.38 (s, 2H), 1.92 (s, 3H), 1.46 (s , 9H).

(3) Boc-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl)

To a suspension of Boc-NH-CH ₂ - (2- (amidino) -5-pyridinyl) × HOAc (0.96 g; 3.1 mmol; see step (2) above) in 75 ml of THF was added a solution of potassium carbonate (1 07 g; 7.7 mmol) and Teos-p-nitrophenyl carbonate (1.14 g; 4.02 mmol) in 15 ml of water. The mixture was stirred overnight. An excess of glycine and potassium carbonate was added and the reaction was continued for 2 hours. THF was evaporated and the residue was extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification.

¹ H NMR (500 MHz; CDCl ₃ ) δ 9.31 (broad, 1H), 8.52 (s, 1H), 8.41 (d, 1H), 8.35 (broad, 1H), 7.74 (d, 1H), 4.97 (broad, 1H), 4.39 (m, 2H), 4.26 (m, 2H), 1.46 (s, 9H), 1.14 (m, 2H), 0.07 (s, 9H).

(4) H ₂ N-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl) × 2HCl

Boc-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl-5-pyridinyl) (0.23 g; 0.58 mmol; see step (3) above) was dissolved in 25 ml of EtOAc saturated with HCl (g) and stirred for 30 minutes The solvent was evaporated and the product was used without further purification Yield 0.21 g (98%).

¹ H NMR (500 MHz; D ₂ O) δ 8.89 (s, 1H), 8.25 (s, 2H), 4.55 (m, 2H), 4.42 (s, 2H), 1.20 (m, 2H), 0.09 (s , 9H).

(5) Boc-Aze-HN-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl)

To a solution of H ₂ N-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl) × 2HCl (0.21 g; 0.57 mmol; see step (4) above), Boc-Aze-OH (0.127 g; 0.631 mmol) and TBTU (233 mg; 0.726 mmol) in 5 ml of DMF was added dimethylaminopyridine (269 mg; 2.20 mmol). The mixture was stirred overnight, poured into 100 ml of water and extracted with EtOAc three times. The combined organic phase was washed with aqueous sodium bicarbonate and water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc to give 170 mg (56%) of the desired product.

¹ H NMR (500 MHz; CDCl ₃ ) δ 9.33 (broad, 1H), 8.54 (s, 1H), 8.41 (d, 1H), 8.36 (broad, 1H), 7.75 (m, 1H), 4.72 (m, 1H), 4.56 (m, 2H), 4.26 (m, 2H), 3.93 (m, 1H), 3.80 (m, 1H), 2.6-2.4 (m, 2H), 1.42 (s, 9H), 1.14 (m , 2H), 0.07 (s, 9H).

(6) H-Aze-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl) × 2HCl

Boc-Aze-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl) (170 mg; 0.356 mmol; see step (5) above) was dissolved in 25 ml of EtOAc saturated with HCl (g), and stirred for 30 minutes The solvent was evaporated and the product was used without further purification. Yield 160 mg (100%).

¹ H NMR (500 MHz; CD ₃ OD) δ 9.00 (m, 1H), 8.84 (m, 1H), 8.23 (d, 2H), 8.10 (m, 1H), 5.09 (m, 1H), 4.7-4.6 (m, 2H), 4.51 (m, 2H), 4.14 (m, 1H), 3.97 (m, 1H), 2.86 (m, 1H), 2.58 (m, 1H), 1.22 (m, 2H), 0.11 ( s, 9H).

(7) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl)

To a solution of H-Aze-NH-CH ₂ - (2- (amino (trimethylsilylethylimino) methyl) -5-pyridinyl) × 2HCl (160 mg; 0.462 mmol; see step (6) above), Ph (3-Cl) ( 5-OCHF ₂ ) - (R) CH (OH) C (O) OH (131 mg; 0.462 mmol; see Example 1 (8) above) and RuBOP (263 mg; 0.505 mmol) in 5 ml of DMF was added diisopropylethylamine (0 30 ml; 1.71 mmol). The mixture was stirred overnight, poured into 100 ml of water and extracted with EtOAc three times. The combined organic phase was washed with aqueous sodium bicarbonate and water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in EtOAc / MeOH (95: 5) to give 148 mg (52%) of the desired product.

(8) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - (2- (methoxyamino (trimethylsilylethylimino) methyl) -5-pyridinyl )

Suspension Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) -C (O) -Aze-NH-CH ₂ - (2- (methoxyamino (trimethylsilylethylimino) methyl) -5-pyridinyl) (148 mg; 0.242 mmol; see step (7) above) and O-methylhydroxylamine (202 mg; 2.42 mmol) in 10 ml of acetonitrile was heated at 70 ° C for 3 hours. The mixture was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude material was flash chromatographed on silica gel in EtOAc / MeOH (95: 5) to give 44 mg (28%) of pure material.

¹ H NMR (500 MHz; CDCl ₃ ) δ 8.55 (m, 1H), 8.05 (bt, 1H), 7.70 (m, 1H), 7.58 (s, 1H), 7.56 (d, 1H), 7.22 (m, 1H), 7.16 (m, 1H), 7.03 (m, 1H), 6.50 (t, 1H), 4.92 (s, 1H), 4.89 (m, 1H), 4.55-4.45 (m, 2H), 4.38 (broad , 1H), 4.2-4.1 (m, 3H), 4.00 (s, 3H), 3.73 (m, 1H), 2.69 (m, 1H), 2.44 (m, 1H), 0.97 (m, 2H), 0.02 ( s, 9H).

(9) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((2-methoxyamidino) -5-pyridinyl)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - (2- (methoxyamino (tri-methylsilylethylimino) methyl) -5-pyridinyl) ( 44 mg; 0.069 mmol; see step (8) above) was dissolved in 2 ml of TFA and allowed to react for 1 h. The TFA was evaporated and the residue was partitioned between EtOAc and aqueous sodium bicarbonate. The aqueous layer was extracted with EtOAc and the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 30 mg (88%). Purity> 95%.

¹ H NMR (500 MHz; CDCl ₃ ) δ 8.44 (m, 1H), 8.03 (bt, 1H), 7.91 (m, 1H), 7.60 (m, 1H), 7.19 (m, 1H), 7.13 (m, 1H), 7.00 (m, 1H), 6.52 (t, 1H), 5.6-5.45 (broad, 2H), 4.90 (s, 1H), 4.89 (m, 1H), 4.55-4.4 (m, 2H), 4.27 (broad 1H), 4.12 (m, 1H), 3.92 (s, 3H), 2.68 (m, 1H), 2.41 (m, 1H).

¹³ C-NMR (100 MHz; CDCl ₃ ) (carbonyl and / or amidine carbons) δ 173.0, 170.9, 152.6.

APCI-MS: (M + 1) = 498/500 m / z.

Example 36

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((5-amidino) -2-pyrimidinyl)

(1) 2-amino-2-iminoethylcarbamate · AcOH

N-Boc-aminoacetonitrile (40.2 g; 257.4 mmol) and N-acetylcysteine (42.0 g; 257.4 mmol) were dissolved in methanol (300 ml) at 60 ° C and ammonia was passed over 18 hours. The solvent was removed in vacuo. After ion exchange chromatography (Amberlit IRA-400 (AcOH)) and recrystallization from acetone, 28.4 g (53%) of the subtitle compound was obtained as a white solid.

¹ H NMR (300 MHz; CD ₃ OD) δ 4.41 (t, J = 4.9 Hz, 1 H), 4.01 (s, 2H), 2.91 (d, J = 5.0 Hz, 2H), 2.01 (s, 3H) 1.46 (s, 9H).

(2) 1,3-Bis (dimethylamino) -2-cyanotrimethine perchlorate

A solution of 3-dimethylaminoacrylonitrile (25.0 g; 260.0 mmol) in chloroform (75 ml) was added dropwise to a solution of (chloromethylene) dimethylammonium chloride (50.0 g; 390.1 mmol) in chloroform (175 ml) at 0 ° FROM. The reaction mixture was stirred an additional 2 hours at 0 ° C, then allowed to warm to room temperature overnight, then was heated for 8 hours at reflux temperature. The solvent was removed in vacuo. The residue was added to a mixture of sodium perchlorate (110 mg; 0.898 mmol) in water (150 ml) and ethanol (300 ml). The mixture was heated at reflux for 15 minutes, then cooled and left to stand overnight in the refrigerator. The precipitate was collected and recrystallized from ethanol to obtain 23.8 g (52%) of the subtitle compound as colorless needle crystals.

Mp 140-141 ° C.

¹ H NMR (300 MHz; CDCl ₃ ) δ 8.24 (s, 2H), 3.59 (s, 6H), 3.51 (s, 6H).

(3) Boc-NH-CH ₂ - (5-cyano) -2-pyrimidine

A mixture of tert-butyl 2-amino-2-iminoethylcarbamate · AcOH (5.0 g; 23.8 mmol; see step (1) above) and 1,3-bis (dimethylamino) -2-cyanotrimethine perchlorate (6.0 g ; 23.8 mmol; see step (2) above) in pyridine (300 ml) was stirred under nitrogen at 70-75 ° C for 16 hours and then heated at reflux for 6 hours. The mixture was cooled to room temperature and the solvent was removed in vacuo. The residue was extracted with a hot mixture (1: 1) of ethyl acetate and chloroform, filtered through a small pad of silica and concentrated to give a crude product. Flash chromatography on silica eluting with chloroform afforded 4.0 g (71%) of the subtitled compound as a colorless oil, which solidified upon standing.

Mp 86-87 ° C.

R _f = 0.77 (silica, ethyl acetate / chloroform (3: 2)).

¹ H NMR (300 MHz; DMSO-d ₆ ) δ 9.25 (s, 2H), 7.39 (bt, 1H), 4.39 (d, J = 6 Hz, 2H), 1.38 (s, 9H).

¹³ C NMR (750 MHz; DMSO-d ₆ ) δ 170.4, 160.3, 155.8, 115.2, 106.9, 80.0, 46.3, 28.1.

APCl-MS: (M + 1) = 235 m / z.

(4) Boc-Aze-NH-CH ₂ - ((5-cyano) -2-pyrimidinyl)

Boc-NH-CH ₂ - (5-cyano) -2-pyrimidine (1.14 g; 4.87 mmol; see step (3) above) was dissolved in 50 ml of EtOAc saturated with HCl (g), allowed to react for 1 h and concentrated. The residue was dissolved in 20 ml of DMF and cooled in an ice bath. Diisopropylethylamine (3.5 ml; 0.020 mol), Boc-Aze-OH (1.08 g; 5.37 mmol) and HATU (2.80 g; 5.38 mmol) were added, and the reaction mixture was stirred at room temperature in overnight. The solvent was evaporated and the product was purified by preparative RPLC using CH ₃ CN / 0.1 M NH ₄ OAc (40:60). Acetonitrile was evaporated and the aqueous layer was extracted three times with EtOAc. The combined organic layer was dried (MgSO ₄ ) and evaporated. Yield 1.12 g (72%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 8.95 (s, 2H), 4.82 (d, 2H), 4.74 (m, 1H), 3.95 (m, 1H), 3.84 (m, 1H), 2.6-2.4 ( m, 2H), 1.47 (s, 9H).

(5) Boc-Aze-NH-CH ₂ - ((5-amidino) -2-pyrimidinyl) × HOAc

Solution Boc-Aze-NH-CH ₂ - ((5-cyano) -2-pyrimidinyl) (0.83 g; 2.6 mmol; see step (4) above), N-acetylcysteine (0.43 g; 2 , 6 mmol) and ammonium acetate (0.60 g; 7.8 mmol) in 10 ml of methanol was heated at 60 ° C in a nitrogen atmosphere for 2 days. The solvent was evaporated and the crude material was purified by preparative RPLC using a gradient of CH ₃ CN / 0.1 M NH ₄ OAc (5:95 to 100: 0). Fractions of interest were freeze dried to give 1.0 g (93%) of the desired material.

¹ H NMR (300 MHz; D ₂ O; signals blocked by the signal from HDO) δ 9.17 (s, 2H), 4.1-3.9 (m, 2H), 2.60 (m, 1H), 2.29 (m, 1H), 1.93 (s, 3H), 1.44 (s, 9H).

(6) Boc-Aze-NH-CH ₂ - [(5- (amino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl]

To a suspension of Boc-Aze-NH-CH ₂ - ((5-amidino) -2-pyrimidinyl) × HOAc (0.95 g; 2.41 mmol; see step (5) above) in 50 ml THF was added a solution of Teos- p-nitrophenyl carbonate (0.85 g; 3.0 mmol) and potassium carbonate (1.0 g; 7.2 mmol) in 10 ml of water. The mixture was stirred for 24 hours, concentrated and partitioned between water and methylene chloride. The organic layer was washed twice with saturated aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel in heptane / EtOAc (1: 1). Yield 1.04 g (90%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 9.16 (s, 2H), 4.80 (d, 2H), 4.73 (m, 1H), 4.26 (m, 2H), 4.0-3.8 (m, 2H), 2.6- 2.4 (m, 2H), 1.47 (s, 9H), 1.12 (m, 2H), 0.07 (s, 9H).

(7) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5- (amino (trimethylsilylethylimino) methyl)) - 2- pyrimidinyl]

Boc-Aze-NH-CH ₂ - [(5- (amino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl] (0.209 g; 0.437 mmol; see step (6) above) was dissolved in 25 ml of EtOAc saturated with HCl (g ), and allowed to interact for 15 minutes. The solvent was evaporated and the residue was dissolved in 4 ml of DMF. Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (0.100 g; 0.396 mmol; added Example 1 (8) above), RuBOP (0.231 g; 0.444 mmol) were added. and diisopropylethylamine (0.208 g; 1.61 mmol) and the mixture was stirred for 80 minutes. The reaction mixture was poured into 100 ml of water and extracted with EtOAc three times. The combined organic layer was washed with brine, dried (Na ₂ SO ₄ ) and evaporated. The crude product was purified by preparative RPLC using a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (1: 1). Yield 63 mg (26%).

¹ H NMR (400 MHz; CDCl ₃ ; mixture of rotamers) δ 9.3 (broad, 1H), 9.03 (s, 2H, minor rotamer), 9.00 (s, 2H, main rotamer), 8.25 (m, 1H, main rotamer) 7.9 (broad, 1H), 7.80 (m, 1H, minor rotamer), 7.2-6.9 (m, 3H), 6.50 (t, 1H), 5.14 (s, 1H, minor rotamer), 5.08 (m, 1H, minor rotamer), 4.94 (s, 1H, main rotamer), 4.80 (m, 1H, main rotamer), 4.7-4.4 (m, 2H), 4.3-3.9 (m, 3H), 3.74 (m, 1H, main rotamer ), 2.7-2.1 (m, 2H), 1.03 (m, 2H), 0.01 (s, 9H).

(8) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5-amidino) -2-pyrimidinyl) × TFA

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5- (amino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl] ( 21 mg; 0.034 mmol; see step (7) above) was dissolved in 0.5 ml of methylene chloride and cooled in an ice bath. TFA (2 ml) was added, the mixture was stirred for 60 minutes and then concentrated. The product was freeze dried from water and acetonitrile. Yield 20 mg (100%). Purity 100%.

¹ H NMR (400 MHz; CD ₃ OD; mixture of rotamer signals blocked by the signal from HDO) δ 9.08 (s, 2H), 7.4-7.1 (m, 3H), 6.88 (t, 1H, main rotamer), 6.85 (t , 1H, minor rotamer), 5.30 (m, 1H, minor rotamer), 5.22 (s, 1H, minor rotamer), 5.20 (s, 1H, main rotamer), 4.73 (m, 1H, main rotamer), 4.34 (m , 1H, rotamer), 4.21 (m, 1H, rotamer), 4.15-3.95 (m, 2H, rotamers), 2.73 (m, 1H, rotamer), 2.57 (m, 1H, rotamer), 2.45-2.25 (m, 2H, rotamers).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 173.0, 172.6, 172.1, 171.0, 163.4.

APCl-MS: (M + 1) = 469/471 m / z.

Example 37

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((5-methoxyamidino) -2-pyrimidinyl)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5-methoxyamino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl ]

Suspension Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5-amino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl] ( 40 mg; 0.065 mmol; see example 36 (7) above) and O-methylhydroxylamine (33 mg; 0.40 mmol) in 3 ml of acetonitrile was heated at 70 ° C for 3 hours. The mixture was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 33 mg (79%).

¹ H NMR (400 MHz; CDCl ₃ ; mixture of rotamers) δ 8.76 (s, 2H, main rotamer), 8.70 (s, 2H, rotamer), 8.18 (m, 1H), 7.62 (s, 1H), 7.4-6.9 (m, 4H), 6.50 (bt, 1H), 5.3-4.5 (m, 4H), 4.2-4.05 (m, 3H), 3.96 (s, 3H), 3.68 (m, 1H), 2.8-2.2 (m , 2H), 2.1 (broad, 1H), 0.96 (m, 2H), 0.01 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - ((5-methoxyamidino) -2-pyrimidinyl)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NH-CH ₂ - [(5-methoxyamino (trimethylsilylethylimino) methyl)) - 2-pyrimidinyl] (33 mg; 0.052 mmol; see step (1) above) was dissolved in 0.5 ml of methylene chloride and cooled in an ice bath. TFA (2 ml) was added, the mixture was stirred for 2 hours and then concentrated. The product was freeze dried from water and acetonitrile. Yield 31 mg (81%). Purity 100%.

¹ H NMR (400 MHz; CD ₃ OD; mixture of rotamers, signals blocked by the signal from HDO) δ 8.96 (s, 2H, rotamer), 8.94 (s, 2H, rotamer), 7.4-7.3 (m, 1H), 7.2- 7.1 (m, 2H), 6.88 (t, 1H, rotamer), 6.85 (t, 1H, rotamer), 5.29 (m, 1H, rotamer), 5.24 (s, 1H, rotamer), 5.20 (s, 1H, rotamer ), 4.75-4.55 (m, 2H), 4.33 (m, 1H, rotamer), 4.19 (m, 1H, rotamer), 4.15-3.95 (m, 2H, rotamers), 3.88 (s, 3H, rotamer), 3.86 (s, 3H, rotamer), 2.72 (m, 1H, rotamer), 2.56 (m, 1H, rotamer), 2.45-2.25 (m, 2H, rotamers).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 172.8, 172.6, 172.1, 171.8, 167.8, 167.7, 155.1, 152.3, 152.1.

APCl-MS: (M + 1) = 499/501 m / z.

Example 38

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (3-F)

(1) 2-Fluoro-4-vinylbenznitrile

A solution of 4-bromo-2-fluorobenznitrile (4.92 g; 0.0246 mol), vinyl tributyltin (0.78 g; 0.246 mol) and tetrakistriphenylphosphine (0.67 g; 0.58 mmol) in 250 ml of toluene was refluxed refrigerator under nitrogen overnight. The solvent was evaporated and the residue was flash chromatographed on silica gel in a solvent system from heptane / CH ₂ Cl ₂ (1: 1) to pure CH ₂ Cl ₂ . A colorless oil was obtained which crystallizes. Yield 3.0 g (82%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.56 (m, 1H), 7.3-7.2 (m, 2H), 6.69 (m, 1H), 5.89 (d, 1H), 5.51 (d, 1H).

(2) 2-Fluoro-4-hydroxymethylbenznitrile

A cooled solution (-78 ° C) of 2-fluoro-4-vinylbenznitrile (1.3 g; 8.8 mmol; see step (1) above) in 40 ml of CH ₂ Cl ₂ and 5 ml of methanol were bubbled with ozone (50 l / h; 29 g / m ³ ) for 30 minutes To remove excess ozone, the mixture was then bubbled with argon. Sodium borohydride (0.67 g; 0.018 mol) was added and the cooling bath was removed. The mixture was stirred and the reaction allowed to proceed for 1 h. The mixture was evaporated and 2 M HCl was added. The mixture was extracted twice with diethyl ether and the combined ether fraction was dried (Na ₂ SO ₄ ) and evaporated. The crude product crystallized. Yield 1.1 g (81%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.59 (m, 1H), 7.3-7.2 (m, 2H), 4.79 (d, 2H), 2.26 (t, 1H).

(3) 4-cyano-3-fluorobenzylmethanesulfonate

2-Fluoro-4-hydroxymethylbenznitrile (1.3 g; 8.6 mmol; see step (2) above) was dissolved in 50 ml of CH ₂ Cl ₂ and cooled in an ice bath. Triethylamine (0.87 g; 8.6 mmol) and methanesulfonyl chloride (0.99 g; 8.7 mmol) were added. After stirring for 1.5 hours, the reaction mixture was washed with 1 M HCl. The organic phase was dried (Na ₂ SO ₄ ) and evaporated. The product could be used without purification. Colorless oil yield 1.8 g (92%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.66 (m, 1H), 7.35-7.3 (m, 2H), 5.26 (s, 2H), 3.07 (s, 3H).

(4) Azidomethyl-2-fluorobenznitrile

To an ice-cooled solution of 4-cyano-3-fluorobenzylmethanesulfonate (1.8 g; 7.9 mmol; see step (3) above) was added sodium azide (0.8 g; 0.012 mol). The mixture was stirred overnight and then poured into 200 ml of water and extracted three times with diethyl ether. The combined ether phase was washed five times with water, dried (Na ₂ SO ₄ ) and evaporated. The crude, colorless oil could be used without further purification. Yield 1.2 g (87%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.64 (m, 1H), 7.25-7.18 (m, 2H), 4.47 (s, 2H).

(5) 4-aminomethyl-2-fluorobenznitrile

To a suspension of stannous tin dihydrate chloride (0.45 g; 2.4 mmol) in 20 ml of acetonitrile, thiophenol (1.07 g; 9.7 mmol) and triethylamine (0.726 g; 7.17 mmol) were added with stirring. After this, a solution of 4-azidomethyl-2-fluorobenznitrile (0.279 g; 1.58 mmol; see step (4) above) in several milliliters of acetonitrile was added. After 1.5 hours, the azide was consumed and the solvent was evaporated. The residue was dissolved in methylene chloride and washed three times with 2 M NaOH. The organic phase was extracted twice with 1 M HCl. The combined acidic aqueous phase was washed with methylene chloride and then made basic with 2 M NaOH and extracted three times with methylene chloride. The organic phase was dried (Na ₂ SO ₄ ) and evaporated to give 0.172 g (72%) of the desired subtitled compound, which could be used without purification.

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.58 (m, 1H), 7.3-7.2 (m, 2H), 3.98 (s, 2H), 1.55-1.35 (broad, 2H).

(6) Boc-Aze-NHCH ₂ -Ph (3-F, 4-CN)

To an ice-cooled solution of Boc-Aze-OH (0.194 g; 0.96 mmol) in 5 ml of DMF was added TBTU (0.50 g; 9.6 mmol). After 30 minutes, another solution was added containing 4-aminomethyl-2-fluorobenzonitrile (0.17 g; 0.81 mmol; see step (5) above) and diisopropylethylamine (0.326 g; 2.53 mmol) in 7 ml of DMF. The resulting solution was stirred overnight at room temperature. The solvent was evaporated and the product was purified using preparative RPLC using a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (50:50). Freeze-drying gave 0.237 g (74%) of the desired subtitled compound.

¹ H NMR (300 MHz; CD ₃ OD) δ 7.70 (m, 1H), 7.35-7.25 (m, 2H), 4.65-4.35 (m, 3H), 4.0-3.85 (m, 2H), 2.51 (m, 1H), 2.19 (m, 1H), 1.40 (s, 9H).

(7) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NHCH ₂ -Ph (3-F, 4-CN)

Boc-Aze-NHCH ₂ -Ph (3-F, 4-CN) (0.118 g; 0.354 mmol; from step (6) above) was dissolved in 30 ml of EtOAc saturated with HCl (g). The reaction mixture was stirred for 20 minutes and evaporated. The resulting dihydrochloride and HATU (0.152 g; 0.400 mmol) were dissolved in 5 ml of DMF. This solution was added to an ice-cooled solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (0.101 g; 0.400 mmol; see Example 1 (8) above) in 5 ml DMF. The reaction mixture was stirred overnight at ambient temperature. The solvent was evaporated and the product was purified by preparative OFLC in a mixture of CH ₃ CN / 0.1 M NH ₄ OAc (50:50). Freeze-drying gave 0.130 g (77%) of the desired subtitled compound.

¹ H NMR (500 MHz; CD ₃ OD, mixture of rotamers) δ 7.7-7.6 (m, 1H), 7.35-7.1 (m, 5H), 6.88 (t, 1H, rotamer), 6.86 (t, 1H, rotamer) , 5.25-5.1 (m, 1H plus minor rotamer from the next proton), 4.80 (m, 1H, main rotamer), 4.6-4.4 (m, 2H), 4.36 (m, 1H, main rotamer), 4.18 (m, 1H , main rotamer), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, main rotamer), 2.29 (m, 1H , main rotamer), 2.16 (m, 1H, minor rotamer).

(8) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (3-F)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-NHCH ₂ -Ph (3-F, 4-CN) (0.130 g; 0.278 mmol; see step ( 7) above) was dissolved in 80 ml of ethanol saturated with HCl (g). The mixture was allowed to react at room temperature overnight. The solvent was evaporated and the residue was redissolved in 100 ml of ethanol saturated with NH ₃ (g). The reaction was allowed to proceed slowly at room temperature for two days. The temperature was raised to 50 ° C and the reaction was continued for the next 3 days. After the starting material was consumed, the solvent was evaporated. The product was purified by preparative HPLC and freeze-drying gave 17 mg (13%) of the title compound as its HOAc salt.

¹ H NMR (600 MHz; CD ₃ OD, mixture of rotamers) δ 7.65-7.6 (m, 1H), 7.4-7.3 (m, 3H), 7.25-7.1 (m, 2H), 7.15-6.7 (m, 1H) , 5.25-5.1 (m, 1H plus minor rotamer from the next proton), 4.8 (m, 1H, main rotamer, partially invisible due to CD ₃ OH), 4.6-3.95 (m, 4H), 2.69 (m, 1H, minor rotamer), 2.56 (m, 1H, main rotamer), 2.28 (m, 1H, main rotamer), 2.14 (m, 1H, minor rotamer), 1.90 (s, 3H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 180.6, 173.4, 173.1, 172.9, 164.5, 162.3, 159.8.

APCl-MS: (M + 1) = 485/487 m / z.

Example 39

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF)

(1) 2,6-Difluoro-4 - [(methylsulfinyl) (methylthio) methyl] benznitrile

(Methylsulfinyl) (methylthio) methane (7.26 g; 0.0584 mol) was dissolved in 100 ml of anhydrous THF in an argon atmosphere and cooled to -78 ° C. Butyl lithium in hexane (16 ml; 1.6 M; 0.0256 mol) was added dropwise with stirring. The mixture was stirred for 15 minutes. Meanwhile, a solution of 3,4,5-trifluorobenzonitrile (4.0 g; 0.025 mmol) in 100 ml of anhydrous THF was cooled to -78 ° C in argon atmosphere and the first solution was added via cannula over a period of 35 minutes to the last solution . After 30 minutes, the cooling bath was removed and, when the temperature of the reaction mixture reached room temperature, the reaction mixture was poured into 400 ml of water. THF was evaporated and the remaining aqueous layer was extracted three times with diethyl ether. The combined ether phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 2.0 g (30%).

¹ H NMR (500 MHz; CDCl ₃ ) δ 7.4-7.25 (m, 2H), 5.01 (s, 1H, diastereomer), 4.91 (s, 1H, diastereomer), 2.88 (s, 3H, diastereomer), 2.52 (s , 3H, diastereomer), 2.49 (s, 3H, diastereomer), 2.34 (s, 3H, diastereomer), 1.72 (broad, 1H).

(2) 2,6-Difluoro-4-formylbenznitrile

2,6-Difluoro-4 - [(methylsulfinyl) (methylthio) methyl] benznitrile (2.17 g; 8.32 mmol; see step (1) above) was dissolved in 90 ml of THF and 3.5 ml of concentrated sulfuric acid was added . The mixture was left at room temperature for 3 days and then poured into 450 ml of water. This was followed by a three-time extraction with EtOAc, and the combined ether phase was washed twice with aqueous sodium bicarbonate and brine, dried (Na ₂ SO ₄ ) and evaporated. Yield 1.36 g (98%). The position of the formyl group was determined using ¹³ C NMR. The signal from the fluorinated carbons at 162.7 million ^-1 showed the expected bifurcated form with two coupling constants of the order of 260 Hz and 6.3, respectively, relating to ipso- and meta-bound fluorine atoms.

¹ H NMR (400 MHz; CDCl ₃ ) δ 10.35 (s, 1H), 7.33 (m, 2H).

(3) 2,6-Difluoro-4-hydroxymethylbenznitrile

2,6-Difluoro-4-formylbenznitrile (1.36 g; 8.13 mmol; see step (2) above) was dissolved in 25 ml of methanol and cooled in an ice bath. Sodium borohydride (0.307 g; 8.12 mmol) was added in portions with stirring and the reaction mixture was left for 65 minutes. The solvent was evaporated and the residue was partitioned between diethyl ether and aqueous sodium bicarbonate. The ether layer was washed with additional aqueous sodium bicarbonate and brine, dried (Na ₂ SO ₄ ) and evaporated.

The crude product crystallized quickly and could be used without further purification. Yield 1.24 g (90%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.24 (m, 2H), 4.81 (s, 2H), 2.10 (broad, 1H).

(4) 4-cyano-2,6-difluorobenzylmethanesulfonate

To a solution of 2,6-difluoro-4-hydroxymethylbenznitrile (1.24 g; 7.32 mmol; see step (3) above) and methanesulfonyl chloride (0.93 g; 8.1 mmol) in 60 ml of methylene chloride triethylamine (0.81 g; 8.1 mmol) was added with stirring. After 3 hours at 0 ° C, the mixture was washed twice with 1 M HCl and once with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 1.61 g (89%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.29 (m, 2H), 5.33 (s, 2H), 3.07 (s, 3H).

(5) 4-Azidomethyl-2,6-difluorobenznitrile

A mixture of 4-cyano-2,6-difluorobenzylmethanesulfonate (1.61 g; 6.51 mmol; see step (4) above) and sodium azide (0.72 g; 0.0111 mol) in 10 ml of water and 20 ml of DMF stirred at room temperature overnight. The resulting material was further poured into 200 ml of water and extracted three times with diethyl ether. The combined ether phase was washed five times with water, dried (Na ₂ SO ₄ ) and evaporated. A small sample was evaporated for NMR and the product was crystallized. The residue was carefully evaporated, but not completely dry. The yield (theoretically 1.26 g) was considered completely quantitative based on NMR and analytical HPLC.

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.29 (m, 2H), 4.46 (s, 2H).

(6) 4-aminomethyl-2,6-difluorobenznitrile

This interaction was carried out in accordance with the procedure described in J. Chem. Res. (M) (1992) 3128. To a suspension of 520 mg of 10% Pd / C (50% humidity) in 20 ml of water was added a solution of sodium borohydride (0.834 g; 0.0221 mol) in 20 ml of water. As a result, gas was released. 4-Azidomethyl-2,6-difluorobenznitrile (1.26 g; 6.49 mmol; see step (5) above) was dissolved in 50 ml of THF and added to the water mixture in an ice bath over 15 minutes. The mixture was stirred for 4 hours, after which 20 ml of 2 M HCl was added and the mixture was filtered through Celite. Celite was washed with additional water, the combined aqueous phase was washed with EtOAc, then made basic with 2 M NaOH. This was followed by three times extraction with methylene chloride and the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 0.87 g (80%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.20 (m, 2H), 3.96 (s, 2H), 1.51 (broad, 2H).

(7) 2,6-Difluoro-4-tert-butoxycarbonylaminomethylbenznitrile

A solution of 4-aminomethyl-2,6-difluorobenzonitrile (0.876 g; 5.21 mmol; see step (6) above) was dissolved in 50 ml of THF and di-tert-butyl dicarbonate (1.14 g; 5.22 mmol) was added to 10 ml THF. The mixture was stirred for 3.5 hours. THF was evaporated and the residue was partitioned between water and EtOAc. The organic layer was washed three times with 0.5 M HCl and water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 1.38 g (99%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.21 (m, 2H), 4.95 (broad, 1H), 4.43 (broad, 2H), 1.52 (s, 9H).

(8) Boc-Pab (2,6-diF) (OH)

A mixture of 2,6-difluoro-4-tert-butoxycarbonylaminomethylbenznitrile (1.38 g; 5.16 mmol; see step (7) above), hydroxylamine hydrochloride (1.08 g; 0.0155 mol) and triethylamine (1.57 g; 0.0155 mol) in 20 ml of ethanol was stirred at room temperature for 36 hours. The solvent was evaporated and the residue was partitioned between water and methylene chloride. The organic layer was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 1.43 g (92%).

¹ H NMR (500 MHz; CD ₃ OD) δ 7.14 (m, 2H), 4.97 (broad, 1H), 4.84 (broad, 2H), 4.40 (broad, 2H), 1.43 (s, 9H).

(9) Boc-Pab (2,6-diF) × HOAc

This interaction was carried out in accordance with the method described by Judkins et al., Synth. Comm. (1998) 4351. Boc-Pab (2,6-diF) (OH) (1.32 g; 4.37 mmol; see step (8) above), acetic anhydride (0.477 g; 4.68 mmol) and 442 mg of 10% Pd / C (50% humidity) in 100 ml of acetic acid was hydrogenated at 490 kPa (5 atm) for 3.5 hours. The mixture was filtered through Celite, washed with ethanol and evaporated. The residue was dried by freezing from acetonitrile, water and a few drops of ethanol. The subtitled product could be used without further purification. Yield 0.149 g (99%).

¹ H NMR (400 MHz; CD ₃ OD) δ 7.45 (m, 2H), 4.34 (s, 2H), 1.90 (s, 3H), 1.40 (s, 9H).

(10) Boc-Pab (2.6-diF) (Teoc)

To a solution of Boc-Pab (2,6-diF) × HOAc (1.56 g; 5.49 mmol; see step (9) above) in 100 ml of THF and 1 ml of water was added 2- (trimethylsilyl) ethyl-p- nitrophenyl carbonate (1.67 g; 5.89 mmol). A solution of potassium carbonate (1.57 g; 0.0114 mol) in 20 ml of water was added dropwise over 5 minutes. The mixture was stirred overnight. THF was evaporated and the residue was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride, the combined organic phase was washed twice with aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated. Flash chromatography on silica gel in a mixture of heptane / EtOAc (2: 1) gives 1.71 g (73%) of pure compound.

¹ H NMR (400 MHz; COCl ₃ ) δ 7.43 (m, 2H), 4.97 (broad, 1H), 4.41 (broad, 2H), 4.24 (m, 2H), 1.41 (s, 9H), 1.11 (m, 2H), 0.06 (s, 9H).

(11) Vos-Aze-Pab (2.6-diF) (Theos)

Boc-Pab (2,6-diF) (Teos) (1.009 g; 2.35 mmol; see step (10) above) was dissolved in 50 ml EtOAc saturated with HCl (g). The mixture was left for 10 min, evaporated, dissolved in 18 ml of DMF and then cooled in an ice bath. Boc-Aze-OH (0.450 g; 2.24 mmol), RuBOP (1.24 g; 2.35 mmol) and finally diisopropylethylamine (1.158 g; 8.96 mmol) were added. The reaction mixture was stirred for 2 hours, then poured into 350 ml of water and extracted with EtOAc three times. The combined organic phase was washed with brine, dried (Na ₂ SO ₄ ) and evaporated. Flash chromatography on silica gel in a mixture of heptane / EtOAc (1: 3) gave 1.097 g (96%) of the desired compound.

¹ H NMR (500 MHz; CDCl ₃ ) δ 7.46 (m, 2H), 4.65-4.5 (m, 3H), 4.23 (m, 2H), 3.87 (m, 1H), 3.74 (m, 1H), 2.45- 2.3 (m, 2H), 1.40 (s, 9H), 1.10 (m, 2H), 0.05 (s, 9H).

(12) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (Teoc)

Boc-Aze-Pab (2,6-diF) (Teos) (0.256 g; 0.500 mmol; see step (11) above) was dissolved in 20 ml EtOAc saturated with HCl (g). The mixture was left for 10 minutes, evaporated and dissolved in 5 ml of DMF. Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (0.120 g; 0.475 mmol; see Example 1 (8) above), RuBOP (0.263 g; 0.498 mmol) were added. and finally diisopropylethylamine (0.245 g; 1.89 mmol). The reaction mixture was stirred for 2 hours, then poured into 350 ml of water and extracted with EtOAc three times. The combined organic phase was washed with brine, dried (Na ₂ SO ₄ ) and evaporated. Silica gel flash chromatography in EtOAc afforded 0.184 g (60%) of the desired subtitled compound.

¹ H NMR (400 MHz; CD ₃ OD, mixture of rotamers) δ 7.55-7.45 (m, 2H), 7.32 (m, 1H, main rotamer), 7.27 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H), 6.90 (t, 1H, main rotamer), 6.86 (t, 1H, minor rotamer), 5.15 (s, 1H, main rotamer), 5.12 (m, 1H, minor rotamer), 5.06 (s, 1H, minor rotamer), 4.72 (m, 1H, main rotamer), 4.6-4.45 (m, 2H), 4.30 (m, 1H, main rotamer), 4.24 (m, 2H), 4.13 (m, 1H, main rotamer), 4.04 (m, 1H, minor rotamer), 3.95 (m, 1H, minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, main rotamer), 2.22 (m, 1H, main rotamer), 2.10 (m, 1H, minor rotamer), 1.07 (m, 2H), 0.07 (m, 9H).

(13) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (Teos) (81 mg; 0.127 mmol; see step (12) above) was dissolved in 0.5 ml of methylene chloride and cooled in an ice bath. TFA (3 ml) was added and the reaction mixture was left for 75 minutes. TFA was evaporated and the residue was freeze dried from water and acetonitrile. The crude product was purified by preparative OFLC in a mixture of CH ₃ CN / 0.1 M NH ₄ OAC (35:65) to give 39 mg (55%) of the title compound as its HOAc salt, 99% pure.

¹ H NMR (400 MHz; CD ₃ OD, mixture of rotamers) δ 7.5-7.4 (m, 2H), 7.32 (m, 1H, main rotamer), 7.28 (m, 1H, minor rotamer), 7.2-7.1 (m, 3H), 6.90 (t, 1H, main rotamer), 6.86 (t, minor rotamer), 5.15 (s, 1H, main rotamer), 5.14 (m, 1H, minor rotamer), 5.07 (s, 1H, minor rotamer) , 4.72 (m, 1H, main rotamer), 4.65-4.45 (m, 2H), 4.30 (m, 1H, main rotamer), 4.16 (m, 1H, main rotamer), 4.03 (m, 1H, minor rotamer), 3.95 (m, 1H, minor rotamer), 2.63 (m, 1H, minor rotamer), 2.48 (m, 1H, primary rotamer), 2.21 (m, 1H, primary rotamer), 2.07 (m, 1H, minor rotamer), 1.89 (s, 3H).

¹³ C-NMR (75 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 171.9, 171.2, 165.0, 162.8, 160.4.

APCl-MS: (M + 1) = 503/505 m / z.

Example 40

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (OMe)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (Ohm, Theos)

Mixture of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (Teos) (64 mg; 0.099 mmol; see example 39 ( 12) above) and O-methylhydroxylamine hydrochloride (50 mg; 0.60 mmol) in 4 ml of acetonitrile was heated at 70 ° C for 3 hours. The solvent was evaporated and the residue was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 58 mg (87%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 7.90 (bt, 1H), 7.46 (m, 1H), 7.25-6.95 (m, 5H), 6.51 (t, 1H), 4.88 (s, 1H), 4.83 ( m, 1H), 4.6-4.5 (m, 2H), 4.4-3.9 (m, 4H), 3.95 (s, 3H), 3.63 (m, 1H), 2.67 (m, 1H), 2.38 (m, 1H) , 1.87 (broad, 1H), 0.98 (m, 2H), 0.01 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (OMe)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (OMe, Teos) (58 mg; 0.086 mmol; see step ( 1) above) was dissolved in 3 ml of TFA, cooled in an ice bath and allowed to react for 2 hours. TFA was evaporated and the residue was dissolved in EtOAc. The organic layer was washed twice with aqueous sodium carbonate and water, dried (Na ₂ SO ₄ ) and evaporated. The residue was freeze dried from water and acetonitrile to give 42 mg (92%) of the title compound. Purity 94%.

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.95 (bt, 1H), 7.2-7.1 (m, 4H), 6.99 (m, 1H), 6.52 (t, 1H), 4.88 (s, 1H), 4.85- 4.75 (m, 3H), 4.6-4.45 (m, 2H), 4.29 (broad, 1H), 4.09 (m, 1H), 3.89 (s, 3H), 3.69 (m, 1H), 2.64 (m, 1H) , 2.38 (m, 1H), 1.85 (broad, 1H).

¹³ C-NMR (100 MHz; CDCl ₃ ) (carbonyl and / or amidine carbons) δ 172.1, 169.8, 151.9.

APCl-MS: (M + 1) = 533/535 m / z.

Example 41

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF)

(1) 2,5-Difluoro-4 - [(methylsulfinyl) (methylthio) methyl] benznitrile

(Methylsulfinyl) (methylthio) methane (3.16 g; 0.0255 mol) was dissolved in 50 ml of anhydrous THF in an argon atmosphere and then cooled to -78 ° C. Butyllithium in hexane (16 ml; 1.6 M; 0.0256 mol) was added dropwise using a syringe. The mixture was stirred for 15 minutes. Meanwhile, a solution of 2,4,5-trifluorobenzonitrile (2.0 g; 0.013 mol) in 50 ml of anhydrous THF was cooled to -78 ° C under argon and the first solution was added via cannula over a period of 3-5 minutes last solution. After 30 minutes, the cooling bath was removed and, when the temperature of the reaction mixture reached room temperature, the reaction mixture was poured into 200 ml of water. THF was evaporated and the remaining aqueous layer was extracted three times with diethyl ether. The combined ether phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product began to crystallize and could be used as it is in the next step. Yield 2.8 g (84%).

¹ H NMR (500 MHz; CDCl ₃ ) δ 7.51-7.44 (m, 2H, main diastereomer), 7.39 (dd, 1H, minor diastereomer), 5.00 (s, 1H, minor diastereomer), 4.92 (s, 1H, basic diastereomer), 2.59 (s, 3H, minor diastereomer), 2.56 (s, 1H, primary diastereomer), 2.46 (s, 1H, minor diastereomer), 2.40 (s, 1H, primary diastereomer).

(2) 2,5-Difluoro-4-formylbenznitrile

2,5-Difluoro-4 - [(methylsulfinyl) (methylthio) methyl] benznitrile (2.8 g; 0.0107 mol; see step (1) above) was dissolved in 100 ml of THF and 6.5 g of concentrated sulfuric acid was added . The mixture was left at room temperature for 6 days and then poured into 500 ml of water. This was followed by three times extraction with diethyl ether, the combined ether phase was washed several times with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product was flash chromatographed on silica gel using heptane / EtOAc (8: 2). Yield 1.2 g (67%). The position of the formyl group was determined using ¹³ C NMR. The carbon signals from fluorinated carbons at 160.1 and 158.4, respectively, were doublets, not the quartets they would be if the formyl group were in position 2.

¹ H NMR (300 MHz; CDCl ₃ ) δ 10.36 (d, 1H), 7.72 (dd, 1H), 7.54 (dd, 1H).

(3) 2,5-Difluoro-4-hydroxymethylbenznitrile

2,5-Difluoro-4-formylbenznitrile (3.60 g; 0.0215 mol; see step (2) above) was dissolved in 50 ml of methanol and cooled in an ice bath. Sodium borohydride (0.815 g; 0.0215 mol) was added in portions with stirring, and the reaction mixture was left for 45 minutes. Water (300 ml) was added, and then 2 M HCl was carefully added until an acidic pH was reached. The mixture was extracted three times with diethyl ether, the combined ether phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The crude product crystallized quickly and could be used without further purification. Yield 3.1 g (85%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.45 (dd, 1H), 7.30 (dd, 1H), 4.85 (s, 2H), 2.10 (broad, 1H).

(4) 4-cyano-2,5-difluorobenzylmethanesulfonate

To a solution of 2,5-difluoro-4-hydroxymethylbenznitrile (3.10 g; 0.0183 mol; see step (3) above) and methanesulfonyl chloride (2.21 g; 0.0192 mol) in 60 ml of methylene chloride triethylamine (1.95 g; 0.0192 mol) was added with stirring. After 1.5 hours at 0 ° C., the mixture was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 4.5 g (99%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.45-7.35 (m, 2H), 5.32 (s, 2H), 3.13 (s, 3H).

(5) 4-Azidomethyl-2,5-difluorobenznitrile

A mixture of 4-cyano-2,5-difluorobenzylmethanesulfonate (4.5 g; 0.0182 mol; see step (4) above) and sodium azide (2.0 g; 0.031 mol) in 20 ml of water and 40 ml of DMF was stirred at room temperature for 2 hours. Then it was poured into 300 ml of water and extracted three times with diethyl ether. The combined ether phase was washed several times with water, dried (Na ₂ SO ₄ ) and evaporated. A small sample was evaporated for NMR and the product was crystallized. The residue was carefully evaporated, but not completely dry. The yield (theoretically 3.5 g) was considered completely quantitative based on NMR and analytical HPLC.

¹ H NMR (500 MHz; CDCl ₃ ) δ. 38 (dd, 1H), 7.32 (dd, 1H), 4.54 (s, 2H).

(6) 4-Aminomethyl-2,5-difluorobenznitrile

This interaction was carried out in accordance with the procedure described in J. Chem. Res. (M) (1992) 3128. To a suspension of 300 mg of 10% Pd / C (50% humidity) in 20 ml of water was added a solution of sodium borohydride (0.779 g; 0.0206 mol) in 20 ml of water. As a result, gas was released. 4-Azidomethyl-2,5-difluorobenznitrile (1.00 g; 5.15 mmol; from step (5) above) was dissolved in 60 ml of THF and added to the aqueous mixture in an ice bath. The mixture was stirred for 1.5 hours, after which 10 ml of 2 M HCl was added and the mixture was filtered through Celite. Celite was washed with additional water, the combined aqueous phase was washed with EtOAc, then made basic with 2 M NaOH. This was followed by three times extraction with methylene chloride, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Yield 0.47 g (54%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.39 (dd, 1H), 7.29 (dd, 1H), 3.99 (s, 2H), 1.45 (broad, 2H).

(7) 2,5-Difluoro-4-tert-butoxycarbonylaminomethylbenznitrile

A solution of 4-aminomethyl-2,5-difluorobenzonitrile (0.46 g; 2.7 mmol; see step (6) above) and di-tert-butyl dicarbonate (0.60 g; 2.7 mmol) in 10 ml THF was stirred during the night. THF was evaporated and the residue was partitioned between water and EtOAc. The organic layer was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 0.71 g (97%).

¹ H NMR (300 MHz; CDCl ₃ ) δ 7.35-7.2 (m, 2H), 5.11 (broad triplet, 1H), 4.38 (d, 2H), 1.45 (s, 9H).

(8) Boc-Pab (2.5-diF) (OH)

A mixture of 2,5-difluoro-4-tert-butoxycarbonylaminomethylbenznitrile (0.70 g; 2.6 mmol; see step (7) above), hydroxylamine hydrochloride (0.54 g; 7.8 mmol) and triethylamine (0.79 g; 7.8 mmol) in 10 ml of ethanol was stirred at room temperature for 6 days. Then it was distributed between water and methylene chloride. The aqueous layer was extracted with methylene chloride, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 0.72 g (92%).

¹ H NMR (500 MHz; CD ₃ OD) δ 7.27 (dd, 1H), 7.12 (dd, 1H), 4.29 (s, 2H), 1.47 (s, 9H).

(9) Boc-Pab (2.5-diF) × HOAc

This interaction was carried out in accordance with the method described by Judkins et al., Synth. Comm. (1998) 4351. Boc-Pab (2.5-diF) (OH) (0.70 g; 2.3 mmol; see step (8) above), acetic anhydride (0.25 g; 2.4 mmol) and 230 mg of 10% Pd / C (50% humidity) in 70 ml of acetic acid was hydrogenated at 490 kPa (5 atm) for 2.5 hours. The mixture was filtered through Celite and evaporated. The residue was freeze dried from acetonitrile and water. The product could be used without further purification in the next step. Yield 0.80 g (100%).

¹ H NMR (500 MHz; CD ₃ OD) δ 7.49 (dd, 1H), 7.31 (dd, 1H), 4.33 (s, 2H), 1.91 (s, 3H), 1.46 (s, 9H).

(10) Boc-Pab (2.5-diF) (Teoc)

To a suspension of Boc-Pab (2.5-diF) × HOAc (0.80 g; 2.3 mmol; see step (9) above) in 50 ml of THF was added 2- (trimethylsilyl) ethyl p-nitrophenyl carbonate (0, 85 g; 3.0 mmol). A solution of potassium carbonate (0.80 g; 5.8 mmol) in 10 ml of water was added dropwise. The mixture was stirred overnight. The excess Teos reagent was destroyed by adding glycine (0.100 g) and potassium carbonate (0.75 g) to the solution and allowing it to react for an additional 2 hours. THF was evaporated and the residue was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Flash chromatography on silica gel in a mixture of heptane / EtOAc (2: 1) gives 0.72 g (72%) of pure compound.

¹ H NMR (400 MHz; CDCl ₃ ) δ 8.00 (dd, 1H), 7.15 (dd, 1H), 4.98 (broad, 1H), 4.36 (bd, 2H), 4.24 (m, 2H), 1.45 (s, 9H), 1.12 (m, 2H), 0.07 (s, 9H).

(11) H-Pab (2.5-diF) (Teoc) × 2HCl

Boc-Pab (2.5-diF) (Teos) (0.38 g; 0.88 mmol; see step (10) above) was dissolved in 50 ml EtOAc saturated with HCl (g). The mixture was left for 30 minutes and evaporated.

¹ H NMR (500 MHz; CD ₃ OD) δ 7.75-7.6 (m, 2H), 4.46 (m, 2H), 4.3 (s, 2H), 1.15 (m, 2H), 0.07 (s, 9H) .

(12) Vos-Aze-Pab (2.5-diF) (Theos)

To a stirred solution of Boc-Aze-OH (0.189 g; 0.94 mmol), H-Pab (2.5-diF) (Teos) × 2HCl (0.36 g; 0.89 mmol; see step (11) above) ) and RuBOP (0.54 g; 1.03 mmol) in 5 ml of DMF was added diisopropylethylamine (0.49 g; 3.8 mmol) and the mixture was allowed to react overnight. The resulting product was poured into aqueous sodium bicarbonate and extracted with EtOAc three times. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Flash chromatography on silica gel in a mixture of heptane / EtOAc (3: 7) gave a sufficiently pure compound. Yield 0.25 g (48%).

¹ H NMR (500 MHz; CDCl ₃ ) δ 7.98 (dd, 1H), 7.13 (dd, 1H), 4.69 (m, 1H), 4.53 (m, 2H), 4.22 (m, 2H), 3.92 (m, 1H), 3.79 (m, 1H), 2.55-2.35 (m, 2H), 1.44 (s, 9H), 1.11 (m, 2H), 0.06 (s, 9H).

(13) H-Aze-Pab (2.5-diF) (Teoc) × 2HCl

Boc-Aze-Pab (2.5-diF) (Teos) (0.25 g; 0.49 mmol; see step (12) above) was dissolved in 50 ml of EtOAc saturated with HCl (g). The mixture was left for 30 minutes and evaporated. The product was used in the next step without further purification. Yield 0.23 g (97%).

¹ H NMR (400 MHz; CD ₃ OD) δ 7.59 (dd, 1H), 7.47 (dd, 1H), 5.14 (m, 1H), 4.54 (m, 2H), 4.48 (m, 2H), 4.15 (m , 1H), 3.96 (m, 1H), 2.87 (m, 1H), 2.56 (m, 1H), 1.17 (m, 2H), 0.05 (s, 9H).

(14) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (Teoc)

To a solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) OH (0.12 g; 0.47 mmol; see example 1 (8) above), H-Aze -Pab (2.5-diF) (Teos) × 2HCl (0.23 g; 0.47 mmol; see step (13) above) and RuBOP (0.27 g; 0.52 mmol) in 10 ml of DMF were added diisopropylethylamine (0.245 g; 1.90 mmol) and the mixture was stirred overnight. The resulting product was poured into water and extracted with EtOAc three times. The combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. Flash chromatography on silica gel in EtOAc gave 100 mg of the pure fraction and 30 mg of the fraction with 90% purity. Total yield 0.13 g (41%).

¹ H NMR (400 MHz; CDCl ₃ ) δ 9.80 (broad, 1H), 8.05 (bt, 1H), 7.94 (dd, 1H), 7.20 (m, 1H), 7.2-7.1 (m, 2H), 7.02 ( m, 1H), 6.54 (t, 1H), 4.93 (s, 1H), 4.91 (m, 1H), 4.51 (m, 2H), 4.28 (broad, 1H), 4.23 (m, 2H), 4.13 (m , 1H), 3.74 (m, 1H), 2.69 (m, 1H), 2.43 (m, 1H), 1.73 (broad, 1H), 1.11 (m, 2H), 1.11 (s, 9H).

(15) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5di-F)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (Teoc) (60 mg; 0.093 mmol; pure fraction from step ( 14) above) was dissolved in 3 ml of TFA and left for 1 h at room temperature. TFA was evaporated and the residue was freeze dried from water and acetonitrile to give 55 mg (96%) of the title compound as its TFA salt, purity> 99%.

¹ H NMR (500 MHz; CD ₃ OD, mixture of rotamers) δ 7.55-7.3 (m, 3H), 7.2-7.1 (m, 2H), 6.88 (t, 1H, main rotamer), 6.86 (t, 1H, minor rotamer), 5.22 (m, 1H, minor rotamer), 5.20 (s, 1H, minor rotamer), 5.13 (s, 1H, minor rotamer), 4.80 (m, 1H, main rotamer), 4.6-4.45 (m, 2H ), 4.36 (m, 1H, main rotamer), 4.19 (m, 1H, main rotamer), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer ), 2.54 (m, 1H, main rotamer), 2.28 (m, 1H, main rotamer), 2.14 (m, 1H, minor rotamer).

¹³ C-NMR (75 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, mixture of rotamers) δ 173.0, 172.6, 172.1, 172.0, 162.4.

APCl-MS: (M + 1) = 503/505 m / z.

Example 42

Ph (3-Ch (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (OMe)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (OMe, Teoc)

Mixture of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,6-diF) (Teoc) (40 mg; 0.062 mmol; see example 41 ( 14) above) and O-methylhydroxylamine hydrochloride (58 mg; 0.70 mmol) in 5 ml of acetonitrile was heated at 70 ° C for 2 hours. The solvent was evaporated and the residue was partitioned between water and EtOAc. The aqueous layer was extracted with EtOAc, the combined organic phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The product could be used without further purification. Yield 35 mg (84%).

¹ H NMR (600 MHz; CDCl ₃ ) δ 7.99 (bt, 1H), 7.72 (s, 1H), 7.20 (m, 1H), 7.15-7.1 (m, 1H), 7.07 (dd, 1H), 7.01 ( m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.88 (m, 1H), 4.48 (m, 2H), 4.2-4.1 (m, 3H), 3.95 (s, 3H), 3.67 (m, 1H), 2.68 (m, 1H), 2.41 (m, 1H), 0.97 (m, 2H), 0.07 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2.5-diF (OMe)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (2,5-diF) (OMe, Teoc) (35 mg; 0.052 mmol; see step ( 1) above) was dissolved in 3 ml of TFA and allowed to interact for 30 minutes. TFA was evaporated and the residue was freeze dried from water and acetonitrile to give 29 mg (99%) of the title compound. Purity 97%.

¹ H NMR (300 MHz; CDCl ₃ ) δ 8.01 (bt, 1H), 7.45 (dd, 1H), 7.20 (m, 1H), 7.15 (m, 1H), 7.09 (dd, 1H), 7.02 (m, 1H), 6.54 (t, 1H), 5.2-5.0 (m, 2H), 4.95-4.85 (m, 2H), 4.6-4.4 (m, 2H), 4.25 (broad, 1H), 4.13 (m, 1H) 3.90 (s, 3H), 3.71 (m, 1H), 2.69 (m, 1H), 2.43 (m, 1H).

¹³ C-NMR (75 MHz; CDCl ₃ ) (carbonyl and / or amidine carbons) δ 173.0, 170.9, 152.6.

APCl-MS: (M + 1) = 533/535 m / z.

Example 43

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OEt)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Oet, Theos)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (55 mg; 0.090 mmol; see Example 1 (9) above) and O-ethylhydroxylamine hydrochloride (53 mg; 0.54 mmol) was dissolved in 4 ml of THF. The mixture was stirred at 60 ° C. for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel, eluting with a methylene chloride / methanol mixture (95: 5), which provided 55 mg (93%) of the subtitle compound.

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.84 (bt, 1H), 7.59 (bs, 1H), 7.47 (bd, 1H), 7.29 (bd, 1H), 7.21 (m, 1H), 7.14 ( m, 1H), 7.02 (m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.86 (m, 1H), 4.55-4.4 (m, 2H), 4.25-4.1 (m, 5H) , 3.69 (m, 1H), 2.66 (m, 1H), 2.41 (m, 1H), 1.33 (t, 3H), 0.98 (m, 2H), 0.02 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OEt)

To an ice-cooled solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OEt, Teoc) (55 mg; 0.084 mmol; see step (1) above) in 0.5 ml of methylene chloride was added 3 ml of TFA. The mixture was stirred (ice bath) for 160 minutes. The material was purified using preparative HPLC. The fractions of interest were combined and freeze dried (2x), collecting 20 mg (47%) of the title compound.

¹ H-NMR (400 MHz; CD ₃ OD) rotamers: δ 7.59 (bd, 2H), 7.35 (m, 1H), 7.32 (bd, 2H), 7.25-7.1 (m, 2H), 6.89 (t, 1H , main rotamer), 6.86 (t, 1H, minor rotamer), 5.18 (s, 1H, main rotamer), 5.18 (m, 1H, minor rotamer), 5.11 (s, 1H, minor rotamer), 4.77 (m, 1H ), 4.5-4.3 (m, 3H), 4.2-3.9 (m, 3H), 2.67 (m, 1H, minor rotamer), 2.52 (m, 1H, main rotamer), 2.28 (m, 1H, main rotamer), 2.15 (m, 1H, minor rotamer); 1.28 (t, 3H).

¹³ C-NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 172.4, 171.9, 171.4, 153.8, 152.3.

MS (m / z) 509 (M-1) ^- , 511 (M + 1) ⁺ .

Example 44

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-n-Pr)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-n-Pr, Teoc)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (53 mg; 0.087 mmol; see Example 1 (9) above) and O-n α-propylhydroxylamine hydrochloride (58 mg, 0.52 mmol) was dissolved in 4 ml of THF. The mixture was stirred at 60 ° C. for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel, eluting with a methylene chloride / methanol mixture (95: 5), which afforded 51 mg (88%) of the subtitle compound.

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.84 (m, 1H), 7.59 (bs, 1H), 7.47 (bd, 2H), 7.28 (bd, 2H), 7.21 (m, 1H), 7.14 ( m, 1H), 7.02 (m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.85 (m, 1H), 4.55-4.4 (m, 2H), 4.2-4.05 (m, 5H) , 3.69 (m, 1H), 2.65 (m, 1H), 2.41 (m, 1H), 1.74 (m, 2H), 1.05-0.95 (m, 5H), 0.03 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-n-Pr)

To an ice-cooled solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-n-Pr, Teoc) (51 mg; 0.078 mmol; see step (1) above) in 0.5 ml of methylene chloride was added 3 ml of TFA. The mixture was stirred (ice bath) for 110 minutes. The material was purified using preparative HPLC. The fraction of interest was evaporated and freeze dried, collecting 20 mg (47%) of the title compound.

¹ H-NMR (500 MHz; CD ₃ OD; rotamers) δ 7.61 (bd, 2H), 7.38 (m, 1H), 7.35 (bd, 2H), 7.22 (m, 1H, main rotamer), 7.18 (m, 1H), 7.15 (m, 1H, minor rotamer), 6.92 (t, 1H, main rotamer), 6.89 (t, 1H, minor rotamer), 5.20 (s, 1H, main rotamer), 5.20 (m, 1H, minor rotamer), 4.80 (m, 1H, main rotamer), 4.5-4.4 (m, 2H, including the minor rotamer corresponding to the main at 4.37), 4.37 (m, 1H, main rotamer), 4.18 (m, 1H, main rotamer) 4.09 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.54 (m, 1H, primary rotamer), 2.30 (m, 1H, primary rotamer), 2.18 ( m, 1H, minor rotamer), 1.73 (m, 2H), 1.01 (t, 3H).

¹³ C-NMR (125 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 171.4, 153.8, 152.3.

MS (m / z) 523 (M-1) ^- , 525 (M + 1) ⁺ .

Example 45

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Pr)

(1) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Pr, Teoc)

Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (Teoc) (50 mg; 0.082 mmol; see example 1 (9) above) and O-iso α-propylhydroxylamine hydrochloride (55 mg; 0.49 mmol) was dissolved in 4 ml of THF. The mixture was stirred at 60 ° C. for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel eluting with a methylene chloride / methanol mixture (95: 5), which afforded 46 mg (84%) of the subtitle compound.

¹ H-NMR (400 MHz; CDCl ₃ ): δ 7.84 (m, 1H), 7.57 (bs, 1H), 7.48 (bd, 2H), 7.29 (bd, 2H), 7.21 (m, 1H), 7.14 ( m, 1H), 7.02 (m, 1H), 6.53 (t, 1H), 4.91 (s, 1H), 4.87 (m, 1H), 4.55-4.45 (m, 2H), 4.42 (m, 1H), 4.2 -4.1 (m, 3H), 3.69 (m, 1H), 2.66 (m, 1H), 2.42 (m, 1H), 1.30 (d, 6H), 0.98 (m, 2H), 0.02 (s, 9H).

(2) Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Pr)

To an ice-cooled solution of Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (O-iso-Pr, Teoc) (46 mg; 0.069 mmol; see step (1) above) in 0.5 ml of methylene chloride was added 3 ml of TFA. The mixture was stirred (ice bath) for 150 minutes. The material was purified using preparative HPLC. The fraction of interest was evaporated and freeze dried (2x) to give 22 mg (58%) of the title compound.

¹ H-NMR (400 MHz; CD ₃ OD; rotamers): δ 7.59 (d, 2H), 7.35 (m, 1H), 7.32 (d, 2H), 7.19 (m, 1H, main rotamer), 7.15 (m , 1H), 7.12 (m, 1H, minor rotamer), 6.89 (t, 1H, minor rotamer), 6.86 (t, 1H, minor rotamer), 5.18 (s, 1H, main rotamer), 5.18 (m, 1H, minor rotamer), 5.12 (s, 1H, minor rotamer), 4.78 (m, 1H, main rotamer), 4.5-3.9 (m, 5H), 2.67 (m, 1H, minor rotamer), 2.52 (m, 1H, main rotamer), 2.28 (m, 1H, main rotamer), 2.15 (m, 1H, minor rotamer), 1.26 (d, 6H).

¹³ C NMR (100 MHz; CD ₃ OD) (carbonyl and / or amidine carbons, rotamers) δ 171.9, 171.4, 153.6.

MS (m / z) 523 (M-1) ^- , 525 (M + 1) ⁺ .

Example 46

The title compounds of Examples 3, 6, 9, 10, 13-15, 17, 19, 21, 23, 25, 27, 28, 32, 34, 36, 38, 39 and 41 were tested in accordance with the above Test And it was found that for all of them, the IC ₅₀ TT values are less than 3.5 μM. It was found that the compounds of examples 3, 6, 9, 10, 13.15, 17, 19, 21, 23, 27, 32, 34 and 39 show values less than 0.02 μM; the compounds of examples 25 and 28 are less than 0.03 μM, the compounds of example 14 are less than 0.04 μM and the compounds of examples 38 and 41 are less than 0.15 μM.

Example 47

The title compounds of Examples 3, 6, 13, 15, 17, 19, 21, 23, 25, 27, 28, 32 and 34 were tested in accordance with Test D above and it was found that for all of them, the IC ₅₀ APTT value less than 1 μM.

Example 48

The title compounds of Examples 1, 2, 4, 5, 7, 12, 16, 18, 20, 22, 24, 26, 29, 30, 33 and 43-45 were tested in accordance with Test E above and it was found that they exhibit oral and / or parenteral bioavailability in rats as the corresponding active inhibitor (free amidine).

Example 49

The title compounds of Examples 1, 2, 7, 8, 11, 12, 16, 18, 22, 24, 26, 29, 33, 37, 40, 43, and 45 were tested in accordance with Test G above, and it was found that they turn into the corresponding active inhibitor (free amidine) in the microsomes of the liver of humans and rats.

Abbreviations

Ac = Acetyl

AcOH = acetic acid

APCl = chemical ionization at atmospheric pressure (in relation to MS)

API = atmospheric pressure ionization (in relation to MS)

aq. = water

AUC = area under the curve

Aze = (S) -azetidine-2-carboxylate (unless otherwise specified)

AzeOH = azetidine-2-carboxylic acid

Bn = benzyl

Boc = tert-butyloxycarbonyl

BSA = Bovine Serum Albumin

Bu = butyl

Bzl = benzyl

Cl = chemical ionization (in relation to MS)

day = day

DCC = Dicyclohexylcarbodiimide

DIBAL-H = diisobutylaluminum hydride

DIPEA = Diisopropylethylamine

DMAP = 4- (N, N-dimethylamino) pyridine

DMF = dimethylformamide

DMSO = Dimethyl Sulfoxide

DVT = Deep Vein Thrombosis

EDC = 1- (3-Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

her. = enantiomeric excess

Et = ethyl

ether = diethyl ether

EtOAc = ethyl acetate

EtOH = Ethanol

Et ₂ O = diethyl ether

h = hour (hours)

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

HBTU = [N, N, N ', N'-tetramethyl-O- (benzotriazol-1-yl) uronium hexafluorophosphate]

HCl = hydrochloric acid, gaseous hydrogen chloride or hydrochloride salt (as appropriate)

Hex = Hexanes

HOAc = acetic acid

HPLC = high performance liquid chromatography

LC = liquid chromatography

Me = methyl

MEM = methoxyethoxymethyl

MeOH = methanol

min = minute (s)

MS = mass spectroscopy

MTBE = methyl tert-butyl ether

NADH = nicotinamide adenine dinucleotide, reduced form

NADPH = nicotinamide adenine dinucleotide phosphate, reduced form

NIH = National Institute of Health (USA)

NIHU = thrombin unit of the National Institute of Health

NMR = nuclear magnetic resonance

OAc = Acetate

Pab = p-amidinobenzylamino

H-Pab = p-amidinobenzylamine

Ph = phenyl

Pr = cut

Pro = (S) -proyninyl

RuBOP = (benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate

QF = tetrabutylammonium fluoride

RedAI = sodium bis (2-methoxyethoxy) aluminum hydride

RPLC = reverse phase high performance liquid chromatography

CT = room temperature

SOP = standard operating procedure (s)

TBTU = [N, N, N ', N'-tetramethyl-O- (benzotriazol-1-yl) uronium tetrafluoroborate]

TEA = triethylamine

Theos = 2- (trimethylsilyl) ethoxycarbonyl

TEMPO = 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical

TFA = trifluoroacetic acid

THF = tetrahydrofuran

THP = tetrahydropyranyl

TLC = thin layer chromatography

TMSCl = trimethylsilyl chloride

TMSCN = Trimethylsilyl Cyanide

UV = UV

Z = benzyloxycarbonyl

The prefixes n-, sec-, iso- and tert- have their usual meanings: normal, secondary, iso- and tertiary. The prefix c means cyclo.

Claims (18)

1. A compound selected from Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe); Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OH) or Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab, or a pharmaceutically acceptable salt of any of these compounds. 2. The compound Ph (3-Cl) (5-OCHF ₂ ) - (R) CH (OH) C (O) -Aze-Pab (OMe) according to claim 1 or its pharmaceutically acceptable salt. 3. A pharmaceutical composition for inhibiting thrombin, comprising any of the compounds of claim 1, or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier. 4. The pharmaceutical composition according to claim 3, comprising a compound according to claim 2 or a pharmaceutically acceptable salt thereof in a mixture with a pharmaceutically acceptable adjuvant, diluent or carrier. 5. The pharmaceutical composition according to claim 3 or 4, where the adjuvant, diluent and / or carrier causes an altered release of the compound. 6. The pharmaceutical composition according to claim 3 or 4, which is suitable for oral administration. 7. The pharmaceutical composition according to claim 5 or 6, which is in the form of a variable release system with a gelling matrix containing a hydrophilic gelling component and an active ingredient. 8. The compound according to claim 1 or 2 for use in the treatment of a condition in which thrombin inhibition is required. 9. The compound according to claim 1 or 2 for use in the treatment of a condition in which anticoagulant therapy is indicated. 10. The compound according to claim 1 or 2 for use in the treatment of thrombosis. 11. The compound according to claim 1 or 2 for use as an anticoagulant. 12. The use of a compound according to claim 1 or 2 as an active ingredient for the manufacture of a medicament for the treatment of a condition in which thrombin inhibition is required. 13. The use of a compound according to claim 1 or 2 as an active ingredient for the manufacture of a medicament for the treatment of a condition in which anticoagulant therapy is indicated. 14. The use of claim 13 or 14, wherein the condition is thrombosis. 15. The application of item 13 or 14, where this condition is hypercoagulation in the blood and / or tissues. 16. The use of compounds according to claim 1 or 2 as an active ingredient for the production of anticoagulant. Priority on points and signs: 12/01/2000 according to claim 1, except for the second compound, claim 2, claims 3-16, except for compound 2 of claim 1; 03/19/2001 according to claim 1, except for compounds 1 and 3, claims 3, 5-16, except for compounds 1 and 3 in the order of claim 1.