ES2373834A1 - Difluorobenzyl ethanolamine derivatives with antimicrobial activity - Google Patents

Abstract

Derivatives of difluorobenzyl ethanolamines with antimicrobial activity, belongs to the field of medical chemistry. It refers to a method of synthesis of fluorinated ethanolamines molecules, specifically difluorobenzyl ethanolamines, as well as to these new molecules and their growth inhibitory activity of species of the Mycobacterium and Nocardia genera, pathogenic bacteria that are involved in the development of diseases such as tuberculosis or leprosy, as well as other infections at the lung, cutaneous or central nervous system level.

Description

Difluorobenzyl ethanolamine derivatives with antimicrobial activity

Technical sector of the invention

The present invention belongs to the field of medical chemistry. It refers to a method of synthesis of fluorinated ethanolamines molecules, specifically difluorobenzyl ethanolamines, as well as to these new molecules and their growth inhibitory activity of species of the Mycobacterium and Nocardia genera, pathogenic bacteria that are involved in the development of diseases such as tuberculosis or leprosy, as well as other infections at the lung, cutaneous or central nervous system level.

Background of the invention

The search for new therapeutic agents is a priority in the fight for the control of infectious diseases. 1 Antimicrobials constitute the fundamental basis of the treatment of infectious diseases, such as those caused by Mycobacterium , specifically Mycobacterium tuberculosis, which is the cause of human tuberculosis and responsible for almost three million deaths per year in the world, being also one of the opportunistic pathogens with the highest incidence in patients with HIV +. Studies conducted in recent years have identified the molecular targets of current drugs in use and their most frequent resistance mechanisms. 2 Some of the active compounds against M. tuberculosis (TB) are isoniazid and pyrazinamide , which affect the synthesis of fatty acids by the bacteria, or ethambutol that is involved in the biosynthesis processes of the bacterial cell wall.

one

The new anti-TB agents must act on a target that is essential for the survival of mycobacteria and must be active throughout their growth cycle, both inside and outside human cells during infection. The M. tuberculosis cell envelope is responsible for its relatively small size compared to other bacteria, has hydrophobic properties and is refractory3, so it resists the action of a drug without changing state or being destroyed. Structurally it consists of a peptidoglycan (PG) layer that contains alternating units of N- acetylglucosamine and N- glycolylmuramic acid. The tetrapeptide side chains are linked in carbon number six of the murmic acid residue which in turn is linked via phosphodiester with arabinogalactan (AG). AG is a polymer composed of residues of D-galactofuranosa and D-arabinofuranosa, it is extremely rare in nature, but necessary for the viability of mycobacteria. On the other hand, it is linked to mycolic acids of 70-90 carbon atoms by esterification of arabinose terminal residues. PG, AG and mycolic acids are the three components that make up the cell wall nucleus of the micolyl-arabinogalactan-peptidoglycan (mAGP) complex. The mAGP contains a lipoarabinomannan (LAM) framework that is involved in the virulence and immuno-pathogenesis of TB. It contains a large number of polar and apolar lipids on the outside of the cell wall that are associated and distributed throughout the cell surface. It is very similar to the cell membrane of gram- negative bacteria . Due to its great complexity and low permeability it contributes significantly to resistance against many therapeutic agents.

Ethambutol, a synthetic amino alcohol that has activity against many strains of the Mycobacterium genus, interrupts the biosynthesis of arabinano that is essential for the formation of AG and LAM, inhibiting the arabinosyl transferase enzyme, 4 interferes with the conversion of D - glucose to D- arabinosa for the transformation of arabinose to AG. Arabinosyl transferase is responsible for the formation of the mAGP complex and if its biosynthetic pathway is inhibited, permeability in the cell wall is increased. An increase in permeability increases, and facilitates the passage of molecules into the cytoplasmic membrane. The design of substrates capable of behaving in this way would represent an important step in the new generation of antibacterials.

In recent decades some have been approved drugs for the treatment of tuberculosis, and others are in clinical phase 5 which highlights the urgent need of new therapies to combat this disease.

In the state of the art there are several documents that describe the obtaining of trifluoromethylamines. Thus, Kuduk et al , 6 "Asymmetric addition reactions of Grignard reagents to chiral 2-trifluoromethyl tert-butyl (Ellman) sulfinimine-ethanol adducts" Tetrahedron Letters 2006 , 47 , 2377-2381, describe that the addition of reagents Grignard to the adducts of trifluoromethyl tert -butyl sulfinimine-ethanol allows to protect the trifluoromethylamines with high yield and a diastereoselectivity between good and excellent. However, the result of stereochemical addition is the opposite of what was expected by controlled chelation the transition state.

Philippe, Christine et al . 7 "Synthesis of New Trifluoromethylated Hydroxyethylamine-Based Scaffolds" Eur. J. Org. Chem 2009 , 5215-5223, refer to the synthesis of new trifluoromethyl-hydroexyethylamines by means of epoxides whose ring opening they obtain with amino compounds, including aliphatic and other amines. The results on the comparison of water and fluorinated and non-fluorinated alcohols in their use as solvents are also given. Regioselectivity is observed and the stereochemistry of the compounds is maintained. Specifically, they describe a process for obtaining trifluoromethyl hydroxyethylamine derivatives from epoxides using nitrogen compounds.

Dos Santos, Mickael et al . 8 "Improved Ritter reaction with CF3-containing oxirane for an Access to central units of protease inhibitors" Tetrahedron Letters 2009 , 50 857-85, investigates the influence of fluorinated alcohols on BF_ {3 } .Et_ {2} in Ritter reactions. The trifluoroethanol / BF 3 .Et 2 O system allows access to the central unit of protease inhibitors through the opening of the fluorinated epoxide cycle with several nitriles.

However, there is still a need for develop new drugs functionally different from those currently described. This search for new active substrates has including both natural products and products of origin synthetic. No one is oblivious to the fact that a problem associated with the discovery of a new drug is the need to study large compound libraries using procedures with good resolution.

Object of the invention

Therefore, the present invention relates to the synthesis of new difluorobenzyl ethanolamines represented by the general formula (I) of Figure 1, as well as its application pharmacological as antimicrobials,

2

where:

a) the protecting group R may be benzyloxycarbonyl, tert- butyloxycarbonyl; Y

b) Ar are derivatives of benzene, furan, thiophene or pyridine, which they can independently possess as substituents fluorine, chlorine, bromine, iodine and methoxy.

Detailed description of the invention

The replacement of hydrogen atoms by atoms of fluoride in the design of new active agents is a strategy general in the design of new therapeutic agents9. Specifically, the design of new agents with activity antimicrobial should have as its main objective the synthesis of active substrates with optimal permeability and stability metabolic

Therefore, the present invention relates to the obtaining and biological behavior of difluorobenzyl ethanolamines that has not been explored in the state of the art since the difluorobenzyl grouping represents a metabolically stable system that can improve the pharmacodynamics of potentially active compounds ^ {10} Regarding the method of obtaining, it has been observed that the epoxidation process used in the present invention leads to the formation of the two possible diastereoisomers, which, once formed, can be separated by chromatography, which has allowed the generation of the four possible diastereoisomers, using the two previously generated allylamines and reducing the total number of stages of the synthesis. In addition, because the double bond of alpha-fluorinated allylamines is strongly deactivated, the use of oxone 11 (potassium peroxymethyl sulfate) as an oxidant has allowed the first example of direct epoxidation thereof to be carried out, which It is not possible with other conventional oxidants. Comparing with the procedures described in the works of Philippe et al . 7 it should be noted that they use a different synthetic methodology and that they only investigate the preparation of one of the diastereoisomers while in the present invention using oxone, they have been achieved The two epoxides.

Given these considerations, it has been designed a difluorobenzyl ethanolamine chemo library formula (I) following the general strategy shown in the Scheme 1. The ethanolamine cluster, as shown, could be prepared by regioselective opening of the epoxy ring of a fluorinated epoxyamime II by reaction with benzyl amines. This epoxide, which contains all the necessary carbon skeleton, could Obtained by stereoselective epoxidation of fluorinated allylamines III conveniently functionalized. Considering the possibility of generating allylamines in chiral form, the subsequent epoxidation allows to prepare all possible isomers

Scheme one

3

         \ vskip1.000000 \ baselineskip
      

Given that there is no collected in the bibliography no example of ethanolamine synthesis difluorobenzyl, and that the presence of CF2 groups in benzyl positions improves the metabolic stability of substrates so it could improve the pharmacodynamics of the potentially active compounds, the investigation was initiated by the Preparation of the library shown in Figure 2.

4

         \ vskip1.000000 \ baselineskip
      

Synthesis of difluorobenzyl ethanolamines of the invention

Fluorinated imines are fundamental feelings and interesting for the introduction of nitrogen and fluorine atoms in polyfunctionalized structures. 12,13 In fact, the Trifluoromethylimines have been previously used in the synthesis of protease inhibitors. 14 In the present invention, the convenient step is used as a key step functionalization of allylamine 4 prepared by alkylation stereoselective of difluorobenzyl imines 5.

Although it was initially considered to exploit the tandem protocol for the synthesis of N- PMP allylamine described by the authors of the present application previously15, which employs a chiral equivalent of vinyl anion, the strategy did not provide in preliminary tests the results that They expected. Alternatively, it was studied to apply the diastereoselective synthesis methodology described by Kuduk et al 6, based on the use of chiral N- tert-butylsulfinimines to prepare chiral amines. 14 This procedure avoids the isolation of N -terc- fluorinated butyl sulphimimines, 15 which are very reactive substrates, and has allowed the preparation of chiral difluorobenzyl allylamines with good yields and selectivity, as shown in Scheme 2 below.

         \ vskip1.000000 \ baselineskip
      

Scheme 2

5

The precursor necessary for the synthesis of imines is not commercial, so it was necessary to synthesize it from ethyl benzoylformate 6. The instability of ethyl hemiacetals 7, make it necessary to directly transform it into hemiaminals. Reaction with ( R ) - tert- butanesulfinamide in the presence of TiOEt 4 at 70 ° C, overnight, provided hemiamine 8, as a 3: 1 mixture of the two possible diastereoisomers, (53% overall yield for the sequence of four steps from 6). The unequivocal separation and allocation of the stereochemistry of both epimers was irrelevant, since both lead to the corresponding difluorobenzyl allylamine 4 with 76% yield when treated with two equivalents of low magnesium vinyl bromide in THF. 18 The compound 4 'was prepared using the same procedure and with similar yields using in this case the (S) -terc- butanesulfinamide, as shown in Scheme 3 below.

         \ vskip1.000000 \ baselineskip
      

Scheme 3

6

7

         \ vskip1.000000 \ baselineskip
      

This strategy provides the 4 and 4 'chiral difluorobenzyl allylamines key to the present invention, with a high enantiomeric purity, 20: 1 dr, as was deduced from the 19 F nuclear magnetic resonance spectrum analysis. The absolute stereochemistry was proposed assuming the model proposed by Kuduk et al . 6, although this end was subsequently confirmed by determining the X-ray structure of brosylate crystal 9 obtained from allylamine 4, as it was shown in Scheme 4 and Figure 2.

8

         \ vskip1.000000 \ baselineskip
      

The next necessary stage was hydrolysis of the chiral auxiliary due to its incompatibility in the conditions epoxidation Hydrolysis using hydrochloric acid (4 M) in dioxane, 19 followed by the reaction of the ammonium salt resulting with base and capture of the free amine with chloride benzyloxycarbonyl provided carbamate 10 with 90% of yield while when the reaction was carried out with Boc 2 O compound 11 was obtained in 82% yield (Scheme 5). Attempts to alternatively also introduce a acetate group were unsuccessful due to its lability in The chromatographic purification process.

         \ vskip1.000000 \ baselineskip
      

Scheme 5

9

         \ vskip1.000000 \ baselineskip
      

Fortunately, epoxidation of difluorobenzyl allylamine 10 with trifluoromethylmethyldioxyran in heterogeneous medium 20 gave a 74% yield an equimolecular mixture (deduced from its 1 H NMR) from the two isomeric epoxides 12a and 12b that were separated by column chromatography on silica gel The application of the same reaction conditions on carbamate 11 yielded the mixture of epoxides 13a and 13b although with a somewhat lower yield. It should be noted at this point that both mixtures showed some instability in the column chromatography process on silica gel, being more pronounced in the case of the N -Boc carbamate.

With the intention of establishing the stereochemistry of the new stereogenic centers created, the epimers 12a and 12b were converted independently into their corresponding oxazolidinones 14, by treatment in a first stage with dibenzylamine in isopropanol at 70 ° C, followed by reaction with sodium hydride in tetrahydrofuran at 0 ° C (Scheme 6). Stereochemistry was determined by careful spectroscopic analysis using different NMR techniques, particularly by correlations observed in the heteronuclear nOe (HOESY) 1 H-19 F experiments. Oxazolidininone 14 shows a spatial proximity between the group CF 2 and the atom H 5, therefore, the assigned stereochemistry is 4 R and 5 S. While for oxazolidinone 15, crossing peaks between the CF2 group and the methylene group are observed, which is in accordance with the stereochemistry shown, 4 R and 5 R , subsequently confirmed by an X-ray diffraction study of it, Figure 3.

10

         \ vskip1.000000 \ baselineskip
      

Following the same sequence of reactions applied to the 4 'enantiomeric difluorobenzyl allylamine, the derived N -Cbz and N -Boc carbamates, compounds 10' and 11 'respectively, were obtained, which were subsequently transformed into the four epoxides 12'a, 12'b , 13'a and 13'b with the yields indicated in Scheme 7.

         \ vskip1.000000 \ baselineskip
      

Scheme 7

eleven

         \ vskip1.000000 \ baselineskip
      

To achieve the proposed objective, it was only necessary to carry out the coupling of the epoxides with the benzylamines to transform them into the objective β-amino alcohols of the work. With the intention of optimizing the process, different opening reaction conditions of epoxide 13b with 3- iodobenzylamine were studied. The best results were when the epoxide was heated to reflux of isopropanol in the presence of the amine to drive 93% yield to the difluorobenzyl amino alcohol 16b. Similarly, epoxide 13a was converted to difluorobenzyl amino alcohol 16a in 87% yield, as indicated in Scheme 8.

         \ newpage
      

Scheme 8

12

The application of the same conditions to epoxides 12, led to the four possible opening isomers 17 with the yields shown in Scheme 9.

Scheme 9

13

14

Finally, the synthesis work was completed following the same protocol as before with the preparation of the chemo library of the difluorobenzyl ethanolamines that are collected in Table 1, with the indicated yields. The structure of the Products obtained are shown below Table 1.

TABLE 1

fifteen

         \ newpage
      

Structures products table 1

17

18

As additional information for a possible study of the relationship between structure and reactivity, it prepared other difluorobenzyl ethanolamines, (27a and 28b) with object of studying on the one hand the influence of the absence of iodine atom and on the other hand the absence of the free NH group of the amine.

         \ newpage
      

Scheme 10

19

         \ vskip1.000000 \ baselineskip
      

Experimental part Synthesis of the compounds

All melting points were measured in a Kofler hot platform. Optical rotations are determined a 5 cm long cuvette and the values of [α] D are expressed in units of 10-1 deg cm 2 g -1. High resolution mass spectra have obtained by electronic impact (EI) at 70 eV, by bombardment fast atom (FAB) or by electrospray ionization (ESI). The 1 H NMR spectra were performed in CDCl 3 at 300 or 400 MHz, and those of 13 C NMR at 75 or 100 MHz. The 1 H spectra they were referenced to residual CHCl 3 (δ 7.26), the 13 C spectra to the central component of the triplet of CDCl 3 to δ 77.0, and the 19 F spectra to the internal reference (CCl_ {F}). The degrees of substitution of Carbon were established by DEPT pulse sequences. A combination of COZY, HMQC and NOE experiments were used when it was necessary for the assignment of chemical shifts of 1 H and 13 C.

Column chromatography refers to flash chromatography and was carried out on silica gel Merck 60, of grain 230-400. All operations involving air sensitive reagents were performed under an inert atmosphere of dry nitrogen or argon, using syringes, oven-dried glassware and freshly dried and distilled solvents.

         \ vskip1.000000 \ baselineskip
      

twenty

         \ vskip1.000000 \ baselineskip
      

Ethyl 2,2-Difluoro-2-phenylacetate . To a solution of ethyl benzoyl formate (2.25 g, 12.0 mmol) dissolved in 8 mL of dry dichloromethane, in a 0 ° C bath, it was added slowly with a 50% deoxofluor addition funnel in toluene (24 mmol). A catalytic amount of absolute ethanol was then added and the reaction mixture was left under continuous stirring at room temperature for 20 h. After confirming by thin layer chromatography the complete disappearance of the starting substrate, hydrolysis was carried out on a water / ice mixture and extraction with dichloromethane. The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The purification of the crude by flash chromatography [ n- hexane: AcOEt (6: 1)] allowed to obtain the product as a transparent liquid, with a yield of 99% (2.4 g). 1 H-NMR (300 MHz, CDCl 3) δ 1.30 (t, J = 7.1 Hz, 3H), 4.30 (c, J = 7.1 Hz, 2H), 7.42-7.53 (m, 3H) , 7.60-7.63 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 12.7, 62.0, 112.4 (t, J CF = 251.7 Hz), 124.4 (t, J CF = 6.0 Hz), 127.6, 129.9, 131.8 (t, J CF = 25.6 Hz), 163.2 t, J CF = 35.4 Hz). 19 F-NMR (282.4 MHz, CDCl 3) δ -104.4 (s, 2F). HRMS (EI +) calculated for C 10 H 10 O 2 F 2 [M +]: 219.0262, found: 219.0263.

         \ vskip1.000000 \ baselineskip
      

twenty-one

2,2-Difluoro-2-phenylethanol . To a suspension of LiAlH 4 (319 mg, 8.4 mmol) in THF (26 mL) at 0 ° C was added, dropwise with an addition funnel, a solution of 2-phenyl-2,2-difluoroacetate of ethyl (560 mg, 2.8 mmol) in 30 mL of THF. It was then left under stirring until the disappearance of the starting substrate by CCF. Then the reaction was treated with Na 2 SO 4 • H2O, the mixture obtained was filtered, washing repeatedly with dichloromethane, and the filtrate was concentrated. The resulting clear oil was purified by flash chromatography [ n- Hexane: AcOEt (4: 1)], providing the alcohol (438 mg, 96%). 1 H-NMR (CDCl 3, 300 MHz) δ 2.23 (sa, 1H), 3.96 (t, J HF = 13.5 Hz, 2H), 7.43-7.48 (m, 3H), 7.48-7.54 (m, 2H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 66.0 (t, J CF = 32.3 Hz), 120.6 (t, J CF = 243.6 Hz), 125.4 (t, J CF = 6.3 Hz), 128.5, 130.3 (t, J CF = 1.3 Hz), 134.3 (t, J CF = 25.4 Hz). 19 F-NMR (CDCl 3, 282.4 MHz) δ-107.7 (t, J HF = 13.5 Hz, 2F). HRMS (EI +) calculated for C 8 H 8 F 2 O [M +]: 158.0543, found 158.0542.

22

         \ vskip1.000000 \ baselineskip
      

N - [(1 RS ) -1-ethoxy-2-phenyl-2,2-difluoroethyl] ( R ) - t -butylsulfinamide (8) . A solution of DMSO (19.6 mmol) in dichloromethane (6 mL) was added to a solution of oxalyl chloride (9.4 mmol) in 6 mL of dichloromethane at -60 ° C. The mixture was allowed to stir at the same temperature for 5 minutes. After this time, a solution of 2-phenyl-2,2-difluoroethanol (1.23 g, 7.8 mmol) in 21 mL of dichloromethane was added dropwise. The resulting mixture was stirred at -60 ° C for 15 minutes. After that time, a solution of triethylamine (39.3 mmol) in 6 mL of dichloromethane was added. After stirring at the same temperature for another 5 minutes, it was allowed to temper for two hours until reaching room temperature. When the starting substrate was consumed, 15 mL of ethanol was added and left under constant stirring at room temperature for 3 hours. The reaction mixture was concentrated to dryness, obtaining a whitish solid residue that was resuspended in ethyl ether and filtered. The ethereal filtrate was concentrated again under reduced pressure, obtaining a light yellow oil that was used immediately in the next reaction step. In a sealed tube under inert conditions, 1-ethoxy-2-phenyl-2,2-difluoroethanol (1.0 mmol), ( R ) -t- butylsulfinamide (1.0 mmol) and titanium tetraethoxide [Ti (OEt) 4 were added }] (5.0 mmol). The reaction mixture was allowed to stir at 70 ° C overnight. After that time it was diluted with 5 mL of ethyl acetate, treated with ice-water and the resulting mixture was extracted with ethyl acetate. The combined organic phases were washed with brine and dried over anhydrous Na2SO4. The solvent was evaporated under reduced pressure and the yellow oil obtained by 1 H NMR indicated that it was a mixture of the diastereoisomers in a 3: 1 ratio. An analytical amount was purified by flash chromatography, using n- hexane: AcOEt (2: 1) as eluent (161 mg, 53%, 2 steps).

Major Diastereoisomer : Yellow solid, mp: 52-54 ° C (CH 2 Cl 2). [α] D 25 = -59.14 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.15 (t, J = 6.9 Hz, 3H), 1.18 (s, 9H), 3.52 (dc, J = 9.3, 6.9 Hz, 1H) , 3.54 (d, J = 10.2 Hz, 1H), 3.96 (dc, J = 9.3, 6.9 Hz, 1H), 4.72 (ddd, J = 10.2, 8.5, 4.2 Hz, 1H), 7.40-7.44 (m, 3H ), 7.49-7.50 (m, 2H). 13 C-NMR (CDCl 3, 75 MHz) δ 14.6, 22.4, 56.8, 65.0, 89.1 (dd, J CF = 35.2 Hz, J CF = 32.5 Hz), 118.7 (t, J CF = 248.4 Hz), 126.3 (t, J CF = 6.2 Hz), 128.0, 130.2, 133.1 (t, J CF = 25.4 Hz). ^ {19} F-NMR (CDCl 3 {}, 282 MHz) \ delta -111.3 (dd, J = 249.0 {FF} Hz, J = 8.5} {HF Hz, 1F), -104.2 (dd, J FF = 249.0, J HF = 4.2 Hz, 1F). EMAR (FAB) calculated for C 14 H 22 F 22 NO 2 S [M + H +]: 306.1339, found: 306.1349.

Minority diastereoisomer: Yellow solid, mp: 45-47 ° C (CH 2 Cl 2). [α] D 25 = -12.05 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.09 (t, J = 6.9 Hz, 3H), 1.23 (s, 9H), 3.40 (dc, J = 9.3, 6.9 Hz, 1H) , 3.76 (dc, J = 9.3, 6.9 Hz, 1H), 4.05 (d, J = 9.3 Hz, 1H), 4.77 (ddd, J = 10.4, 9.3, 3.1 Hz, 1H), 7.42-7.44 (m, 3H ), 7.55-7.57 (m, 2H). 13 C-NMR (CDCl 3, 75 MHz) δ 14.6, 22.5, 57.1, 64.9. 87.7 (dd, J CF = 36.7 Hz, J CF = 32.2 Hz), 118.6 (t, J CF = 249.2 Hz), 126.5 (t, J CF = 6.4 Hz) , 128.1, 130.2, 133.0 (t, J CF = 25.4 Hz). 19 F-NMR (CDCl 3, 282 MHz) δ -113.0 (dd, J FF = 251.9 Hz, J HF = 10.4 Hz, 1F), -103.0 (dd, J FF = 249.0, J HF = 3.1 Hz, 1F). EMAR (FAB) calculated for C 14 H 22 F 22 NO 2 S [M + H +]: 306.1339, found: 306.1335.

         \ vskip1.000000 \ baselineskip
      

2. 3

N - [(1 SR ) -1-Ethoxy-2-phenyl-2,2-difluoroethyl] ( S ) - t -butylsulfinamide (8 ') . When ( S ) -t- butylsulfinamide was used, similar results were obtained. 1.9 g of alcohol led to a mixture of both hemiaminals which, after chromatography, provided 1.0 g (27%) of the majority diastereoisomer [α] D 25 = +71.07 ( c 1.0, CHCl_ {3}, and 351 mg (10%) of the minor diastereoisomer : [α] D 25 = +16.85 ( c 1.0, CHCl 3).

24

         \ vskip1.000000 \ baselineskip
      

[( R ) -t-Butyl sulphinyl] [(1 R ) -1- (phenyldifluoromethyl) allyl] amine (4) . To a solution of the mixture of hemiamine 8 (2.66 g, 8.7 mmol) in dichloromethane (95 mL) at -60 ° C was added a 1 M solution in THF of vinyl magnesium bromide (21.7 mL). The mixture was allowed to slowly warm to -25 ° C, when the starting substrate had completely disappeared. At this point, the mixture was hydrolyzed with aqueous NH4Cl, allowed to warm to room temperature and extracted with dichloromethane. The combined organic phases were washed with brine and dried over anhydrous Na2SO4. Finally, the solvent was removed under reduced pressure and the residue obtained was chromatographed on SiO2 to provide sulfinamide 4 as a yellowish oil, in a 20: 1 diastereomeric ratio, as determined by 1 H-NMR and 19} F-NMR. [α] D 25 = -101.49 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.19 (s, 9H), 3.57 (d, J = 3.0 Hz, 1H), 4.27-4.38 (m, 1H), 5.32 (d, J = 16.8 Hz, 1H), 5.36 (d, J = 9.9 Hz, 1H), 5.62 (ddd, J = 16.8, 9.9, 7.7 Hz, 1H), 7.40.7.47 (m, 5H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 22.4, 56.0, 63.4 (t, J CF = 28.9 Hz), 120.9 (t, J CF = 249.5 Hz), 122.9, 126.1 (t, J CF = 6.3 Hz), 128.3, 130.3 (t, J CF = 4.0 Hz), 130.4 (t, J CF = 1.7 Hz). 133.7 (t; J CF = 26.4 Hz). ^ {19} F-NMR (CDCl 3 {}, 282.4 MHz) \ delta -107.3 (dd, J = 244.3 {FF} Hz, J = 13.0 {HF} Hz, 1F), -103.4 (dd, J FF = 244.3 Hz, J HF = 7.9 Hz, 1F). EMAR (FAB) calculated for C 14 H 20 F 2 NOS [M + H +]: 288.1234, found: 288.1235.

25

         \ vskip1.000000 \ baselineskip
      

( S ) -t-Butylsulfinyl] [(1 S ) -1- (phenyldifluoromethyl) allyl] amine (4 ') . When 1.35 g of the 8 'hemiamine mixture was used, 1.05 g of allylamine (85%) were obtained after chromatography. [α] D 25 = +105.20 ( c 1.0, CHCl 3).

26

         \ vskip1.000000 \ baselineskip
      

Benzyloxycarbonyl [(1 R ) -1- (phenyldifluoromethyl) allyl] amine (10) . To a solution of allylamine 4 (200 mg, 0.7 mmol) in anhydrous methanol (4.5 mL) was added 1.8 mL of a solution of HCl (4 M) in dioxane, and allowed to stir at room temperature for 2 hours After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 4.5 mL of dioxane. Then, K 2 CO 3 (2.1 mmol), benzoyl chloride (3.5 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.1 mmol) were added successively at room temperature. The reaction mixture was left with continuous stirring at room temperature for 18 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to flash chromatography, using n- hexane: ethyl ether (4: 1) as eluent, obtaining a transparent oil corresponding to allylamine 10, with a 91% yield. Mp: 64-66 ° C (CH 2 Cl 2). [α] D 25 = +21.14 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 4.83-5.00 (m, 1H), 5.08 (s, 1H), 5.08 (s, 2H), 5.29 (d, J = 12.0 Hz, 1H), 5.30 (d, J = 16.2 Hz, 1H), 5.84 (ddd, J = 16.2, 10.2, 5.4 Hz, 1H), 7.30-7.37 (m, 5H), 7.37-7.50 (m, 5H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 58.5 (t, J CF = 29.3 Hz), 67.1, 119.2, 120.7 (t, J CF = 258.4 Hz), 125.7 (t, J CF = 6.3 Hz), 128.0, 128.2, 128.3, 128.5, 130.2 (t, J CF = 1.4 Hz), 130.6 (t, J CF = 2.6 Hz), 134.1 (t, J CF = 25.6 Hz), 136.0, 155.6. 19 F-NMR (CDCl 3, 282.4 MHz) δ -106.9 (dd, 2 J FF = 247.2 Hz, J HF = 12.7 Hz, 1F), -104.5 (dd, J = 247.7 {FF} Hz, J = 9.11 {HF} Hz, 1F). EMAR (EI +) calculated for C 18 H 17 F 2 NO 2 [M +]: 317.1227, found: 317.1229.

Benzyloxycarbonyl [(1 R ) -1- (phenyldifluoromethyl) allyl] amine (10 ') . 1.3 g of 4 'allylamine provided 970 mg of 10' N -Cbz allylamine (68%). [α] D 25 = -9.18 ( c 1.0, CHCl 3).

         \ vskip1.000000 \ baselineskip
      

27

( R ) -1-Phenyl-1,1-difluoro-3-butenyl-2-carbamate tert-butyl (11) . To a solution of allylamine 4 (1.0 g, 3.5 mmol) in anhydrous methanol (21.6 mL) 8.7 mL of a solution of HCl (4 M) in dioxane was added, and allowed to stir at room temperature for 2 hours. After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 21.6 mL of dioxane. Then, K 2 CO 3 (10.4 mmol), di- t- butyl dicarbonate (3.5 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.1 mmol) were added successively at room temperature ). The reaction mixture was left with continuous stirring at room temperature for 52 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to flash chromatography, using n- hexane: ethyl ether (7: 1) as eluent, obtaining a white solid corresponding to allylamine 11 (805 mg, 82%). Mp: 49-51 ° C (CH 2 Cl 2). [α] D 25 = +23.67 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.37 (s, 9H), 4.80-4.85 (m, 1H), 4.82 (sa, 1H), 5.28 (d, J = 9.3 Hz, 1H), 5.29 (d, J = 18.0 Hz, 1H), 5.80-5.89 (m, 1H), 7.40-7.43 (m, 3H), 7.47-7.50 (m, 2H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 27.3, 57.8 (t, J CF = 28.1 Hz), 85.1, 118.8, 120.9 (t, J CF = 249.4 Hz ), 125.7 (t, J CF = 6.2 Hz), 128, 130.0, 130.9 (t, J CF = 2.9 Hz), 134.4 (t, J CF = 25.7 Hz), 146.7. ^ {19} F-NMR (CDCl 3 {}, 282.4 MHz) \ delta -106.5 (dd, J = 251.6 {FF} Hz, J = 10.4 {HF} Hz, 1F), -105.3 (dd, J FF = 251.6 Hz, J HF = 11.3 Hz, 1F). EMAR (ESI +) calculated for C 15 H 19 F 2 NNaO 2 [M + Na +]: 306.1282, found: 306.1284.

( S ) -1-Phenyl-1,1-difluoro-3-butenyl-2-carbamate tert-butyl (11 ') . 1.7 g of 4 'allylamine provided 430 mg (41%) of 11' N- Bac-allylamine. [α] D 25 = -20.78 ( c 1.0, CHCl 3).

         \ vskip1.000000 \ baselineskip
      

28

( R ) -2-Phenyl-2,2-difluoro-1 - [( R ) oxyran-2-yl] benzyl ethylcarbamate (12a) and ( R ) -2-phenyl-2,2-difluoro-1- [ ( S ) benzyl oxyran-2-yl] ethylcarbamate (12b) . To a solution of the allylamine 10 (951 mg, 3.0 mmol) in acetonitrile (26 mL) was added 13 mL of an aqueous solution of Na 2 EDTA • 2H 2 O (4 · 10 -4) M). The solution was cooled to 0 ° C and trifluoromethylmethyl ketone (1.0 mL) was added. Subsequently, on the reaction mixture at 0 ° C, NaHCO 3 (17.8 mmol) and Oxone (5.7 mmol) were added together in a single portion. After 3 h of reaction, the suspension was filtered, the phases were separated and the aqueous phase was extracted with dichloromethane. After removing the solvent in vacuo, the solid residue obtained was subjected to flash chromatography, using n- hexane: ethyl ether as eluent with a polarity gradient (10: 1 to 6: 1), which allowed to obtain 441 mg ( 44%) of 12a and 308 mg (31%) of 12b.

Isomer R, R (12a) . White solid, mp: 68-70 ° C (CH 2 Cl 2). [α] D 25 = +4.38 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 2.36 (dd, J = 4.5, 2.4 Hz, 1H), 2.61 (t, J = 4.5 Hz, 1H), 3.18-3.22 (m, 1H), 4.52 (td, J = 12.0, 9Hz, 1H), 4.97 (d, J = 9.0 Hz, 1H), 5.03 (s, 2H), 7.24-7.54 (m, 10H). 13 C-NMR (CDCl 3, 75.5 MHz) δ δ 42.5, 48.0 (t, J CF = 3.6 Hz), 55.0 (t, J CF = 29.3 Hz), 67.3, 121.1 (t, J CF = 249.2 Hz), 125.6 (t, J CF = 6.4 Hz), 127.9, 128.2, 128.5, 128.5, 130.4, 133.9 (t, J CF = 25.9 Hz), 135.9, 156.0. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.9 (d, J HF = 12.0 Hz, 2F). EMAR (EI +) calculated for C 18 H 17 F 2 NO 3 [M +]: 333.1177, found 333.1182.

Isomer R, S (12b) . White solid, mp: 92-94 ° C (CH 2 Cl 2). [α] D 25 = -8.05 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 2.65-2.76 (m, 2H), 3.11-3.16 (m, 1H), 4.27-4.37 (m, 1H), 4.98-5.15 (m , 1H), 5.06 (s, 2H), 7.26-7.56 (m, 10H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 43.9, 48.9 (t, J CF = 3.4 Hz), 57.8 (t, J CF = 29.3 Hz), 67.4, 120.9 (t, J CF = 247.6 Hz), 125.4 (t, J CF = 6.3 Hz), 128.0, 128.3, 128.6, 128.6, 130.6, 133.8 (t, J CF = 25.9 Hz ), 135.8, 155.7. 19 F-NMR (CDCl 3, 282.4 MHz) δ -106.8 (dd, J FF = 251.7 Hz, J HF = 12.5 Hz, 1F), -104.9 (d, J FF = 251.7 Hz, J HF = 13.4 Hz, 1F). HRMS (EI +) calculated for C 18 H 17 F 2 NO 3 [M +]: 333.1177, found 333.1175.

         \ vskip1.000000 \ baselineskip
      

29

( S ) -2-Phenyl-2,2-difluoro-1 - [( S ) oxyran-2-yl] ethyl benzylcarbamate (12'a) and ( S ) -2-phenyl-2,2-difluoro-1 - [( R ) oxyran-2-yl] benzyl ethylcarbamate (12'b) . 965 mg of 10 'allylamine led to a mixture of both diastereoisomeric epoxides which, after flash chromatography provided 307 mg (30%) of the isomer ( S, S ) (12'a) [α] D 25 = -4.98 ( c 1.0, CHCl3), and 441 mg (44%) of the isomer ( S, R ) (12'b) . [α] D 25 = +13.04 ( c 1.0, CHCl 3).

         \ vskip1.000000 \ baselineskip
      

30

( R ) -2-Phenyl-2,2-difluoro-1 - [( R ) oxiran-2-yl] ethylcarbamate tert- butyl (13a) and ( R ) -2-phenyl-2,2-difluoro-1 - [(S) oxiran-2-yl] ethylcarbamate tert -butyl (13b). To a solution of allylamine 11 (631 mg, 2.23 mmol) in acetonitrile (17 mL) was added 8.4 mL of an aqueous solution of Na 2 EDTA • 2H 2 O (4 · 10 ^ {-4} M). The solution was cooled to 0 ° C and trifluoromethylmethyl ketone (4.5 mL) was added. Subsequently, on the reaction mixture at 0 ° C, NaHCO 3 (35.6 mmol) and Oxone (11.1 mmol) were added together in a single portion. After 3 h of reaction, the suspension was filtered, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with brine and dried over Na2SO4. anhydrous. After removing the solvent in vacuo, the solid residue obtained was subjected to flash chromatography, using n- hexane: ethyl ether as eluent with a polarity gradient (10: 1 to 6: 1), which allowed to obtain 187 mg ( 28%) of 13a and 277 mg (42%) of 13b.

Isomer R, R (13a) . White solid, mp: 65-67 ° C (CH 2 Cl 2). [α] D 25 = +8.16 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.32 (s, 9H), 2.51 (dd, J = 4.5, 2.4 Hz, 1H), 2.70 (dd, J = 4.5, 4.5 Hz, 1H), 3.28-3.29 (m, 1H), 4.78 (d, J = 10.2 Hz, 1H), 7.42-7.45 (m, 3H), 7.50-7.53 (m, 2H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 28.0, 42.4, 48.0 (t, J = 3.2 Hz), 54.3 (t, J = 29.1 Hz), 80.2, 121.2 (t, J = 249.3 Hz), 125.6 (t, J = 6.3 Hz), 128.4, 130.2, 134.2 (t, J = 25.4 Hz), 155.1. 19 F-NMR (CDCl 3, 282.4 MHz) δ -106.0 (dd, J FF = 250.5 Hz, J HF = 11.9 Hz, 1F), -104.4 (dd, J FF = 250.5 Hz, J HF = 11.6 Hz, 1F). HRMS (FAB) calculated for C 15 H 20 F 2 NO 3 [M + H +]: 300.1411, found: 300.1402.

Isomer R, S (13b) . White solid, mp: 112-114 ° C (CH 2 Cl 2). [α] D 25 = -5.6 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.35 (s, 9H), 2.67 (dd, J = 4.5, 2.1 Hz, 1H), 2.73 (dd, J = 4.5, 4.5 Hz, 1H), 3.10-3.20 (m, 1H), 4.85 (d, J = 10.2 Hz, 1H), 7.43-7.45 (m, 3H), 7.50-7.53 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.0, 43.8, 49.0 (t, J = 3.2 Hz), 57.1 (t, J = 25.6 Hz), 80.4, 121.0 (t, J = 247.9 Hz), 125.5 (t, J = 6.3 Hz), 128.4, 130.3, 134.1 (t, J = 25.4 Hz), 154.8. 19 F-NMR (CDCl 3, 282.4 MHz) δ -106.4 (dd, J FF = 251.6 Hz, J HF = 11.6 Hz, 1F), -104.8 (dd, J FF = 251.6 Hz, J HF = 13.8 Hz, 1F). EMAR (FAB) calculated for C 15 H 20 F 2 NO 3 [M + H +]: 300.1411, found: 300.1407.

         \ vskip1.000000 \ baselineskip
      

31

( S ) -2-Phenyl-2,2-difluoro-1 - [( S ) oxiran-2-yl] ethylcarbamate tert- butyl (13'a) and ( S ) -2-phenyl-2,2-difluoro -1 - [( R ) oxyran-2-yl] tert -butyl ethylcarbamate (13'b) . 325 mg of 11 'allylamine led to a mixture of both diastereoisomeric epoxides which, after flash chromatography, provided 35 mg (10%) of the isomer ( S, S ) (13'a) [α] D 25 = -7.32 ( c 1.0, CHCl 3) and 67 mg (20%) of the isomer ( S, R ) (13'b) [α] D 25 = +6.13 ( c 1.0 , CHCl3).

         \ vskip1.000000 \ baselineskip
      

32

         \ vskip1.000000 \ baselineskip
      

(4 R , 5 S ) -5 - ((Dibenzylamino) methyl) -4- (phenyl (difluoro) methyl) -2-oxazolidinone (14) . On a solution of epoxy 12a (25 mg, 0.075 mmol) in anhydrous isopropanol (1 mL), and at room temperature, dibenzylamine (0.375 mmol) was added. The resulting mixture was heated in a sealed tube with constant stirring at 70 ° C for 5 hours. After removing the solvent under reduced pressure, the crude was purified by flash chromatography n- hexane: AcOEt (2: 1) on silica gel to remove excess amine. The crude obtained was dissolved in THF (1 mL) and treated at 0 ° C with NaH (pre-washed with hexane to remove the mineral oil and dried under vacuum) (0.75 mmol). The resulting mixture was allowed to stir at room temperature for 12 hours. After that time, it was hydrolyzed with water, extracted with dichloromethane and the organic phases were washed with brine and subsequently dried over anhydrous Na2SO4. The solvent was evaporated in vacuo and the crude obtained was purified by flash chromatography, using n- hexane: ethyl ether (10: 1), as eluent, oxazolidinone 14 (25.2 mg, 80%) being isolated as a white solid, mp: 132-135 ° C (CH 2 Cl 2). [α] D 25 = -11.08 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 2.60 (dd, J = 14.0, 4.6 Hz, 1H), 2.72 (dd, J = 14.0, 6.3 Hz, 1H), 3.57 (s, 4H), 3.72 (td, J = 11.3, 3.6 Hz, 1H), 4.62 (ddd, J = 6.3, 4.6, 3.6 Hz, 1H), 5.50 (sa, 1H), 7.23-7-35 (m, 12H) , 7.38-7.49 (m, 3H). 13 C-NMR (75.5 MHz, CDCl 3) δ 55.1, 59.4, 60.1 (t, J CF = 31.8 Hz), 75.0, 119.7 (t, J CF = 247.6 Hz ), 125.5 (t, J CF = 6.3 Hz), 127.3, 128.4, 128.8, 129.2, 130.8, 132.5 (t, J CF = 25.6 Hz), 138.6, 158.0. 19 F-NMR (CDCl 3, 282.4 MHz) δ -109.8 (dd, J HF = 10.9, 16.1 Hz, 2F). EMAR (FAB) calculated for C 25 H 24 F 24 N 2 O 2 [M + H +]: 423.1884, found 423.1887.

         \ vskip1.000000 \ baselineskip
      

33

         \ vskip1.000000 \ baselineskip
      

(4 R , 5 R ) -5 - ((Dibenzylamino) methyl) -4- (phenyl (difluoro) methyl) -2-oxazolidinone (15) . Following the same procedure, epoxide 12b led to oxazolidinone 15 with 80%, as a white solid, mp: 146-148 ° C (CH 2 Cl 2).
[α] D 25 = -4.48 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 2.94 (d, J = 14.7 Hz, 1H), 3.01 (dd, J = 14.7, 9.3 Hz, 1H), 3.51 (d, J = 13.8 Hz, 2H), 3.76 (d, J = 13.8 Hz, 2H), 3.95 (dt, J = 18.9 , 7.5 Hz, 1H), 4.81 (ddd, J = 18.9, 7.5, 2.4 Hz, 1H), 5.08 (sa, 1H) 7.13-7-43 (m, 15H). 13 C-NMR (75.5 MHz, CDCl 3) δ 52.0 (t, J CF = 3.5 Hz), 53.1, 58.7, 78.9, 120.2 (t, J CF = 250.3 Hz ), 125.3 (t, J CF = 6.3 Hz), 127.0, 128.2, 128.8, 129.0, 131.0, 133.2 (t, J CF = 25.6 Hz), 139.0, 158.1. ^ {19} F-NMR (CDCl 3 {}, 282.4 MHz) \ delta -108.2 (d, J = 251.3 {FF} Hz, 1F), -102.3 (d, J = 251.3 {FF} Hz, 1F ). HRMS (EI +) calculated for C 25 H 24 F 2 N 2 O 2 [M +]: 422.1805, found: 422.1817.

3. 4

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17a) . To a solution of epoxide 12a (40 mg, 0.12 mmol) in anhydrous isopropanol (0.5 mL) at room temperature 3-iodobenzylamine (0.18 mmol) was added and the mixture was heated in a sealed tube at 70 ° C for 15 hours, with constant agitation, until the disappearance of the starting epoxide by CCF was observed. Then, the solvent was removed under reduced pressure and the crude obtained was purified by flash chromatography, using n- hexane: AcOEt (2: 1) as eluent, the amino alcohol 17a being isolated as a yellowish solid (55 mg, 81%). Mp: 85-87 ° C (CH 2 Cl 2). [α] D 25 = -20.82 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 2.34 (sa, 2H), 2.58 (dd, J = 12.3, 9.3 Hz, 1H), 2.68 (dd, J = 12.3, 4.5 Hz, 1H), 3.67 (s, 2H), 4.01 (dd, J = 9.3, 4.3 Hz, 1H), 4.18 (td, J = 13.7, 10.3 Hz, 1H), 5.04 (d, J = 12.3 Hz, 1H), 5.09 (d, J = 12.3 Hz, 1H), 5.59 (d, J = 10.3 Hz, 1H), 7.03 (t, J = 7.7 Hz, 1H), 7.19 (d, J = 7.7 Hz, 1H), 7.26- 7.60 (m, 13H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 51.4, 52.6, 57.2 (t, J CF = 28.0 Hz), 65.7, 67.1, 94.5, 121.7 (t, J CF = 250.9 Hz), 125.6 (t, J CF = 6.4 Hz), 127.2, 127.9, 128.2, 128.4, 128.5, 128.5, 130.2, 134.6 (t, J CF = 25.0 Hz), 136.2, 136.3, 137.0, 141.9, 156.3. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.7 (dd, J FF = 249.1 Hz, J HF = 14.4 Hz, 1F), -103.4 (dd, J FF = 249.1 Hz, J HF = 13.0 Hz, 1F). EMAR (EI +) calculated for C 25 H 25 F 2 IN 2 O 3 [M +]: 566.0878, found: 566.0865.

         \ vskip1.000000 \ baselineskip
      

35

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17b) . Following the same procedure above, 112 mg of epoxy 12b provided 142 mg of 17b (75%). Yellow solid, mp: 109-111 ° C (CH 2 Cl 2). [α] D 25 = -16.8 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 2.69 (dd, J = 12.6, 5.4 Hz, 1H), 2.75 (dd, J = 12.6, 3.9 Hz, 1H), 3.53 (d, J = 13.5 Hz, 1H), 3.59 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 5.4, 3.9 Hz, 1H), 4.34-4.48 (m, 1H), 4.86 (d, J = 12.3 Hz, 1H), 4.97 (d, J = 12.3 Hz, 1H), 6.25 (d, J = 10.3 Hz, 1H), 6.90 (t, J = 7.8 Hz, 1H), 7.10-7.54 (m, 13H). 13 C-NMR (75.5 MHz, CDCl 3) δ 49.8, 52.1, 58.9 (t, J CF = 28.0 Hz), 66.1, 66.4, 93.5, 120.4 (t, J CF = 248.4 Hz), 124.4 (t, J CF = 6.4 Hz), 126.3, 126.9, 127.1, 127.4, 127.5, 129.2, 129.2, 133.8 (t, J CF = 25.7 Hz), 135.1, 135.2, 136.0, 140.9, 155.7. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.9 (dd, J FF = 250.2 Hz, J HF = 11.6 Hz, 1F), -102.0 (dd, J FF = 250.2 Hz, J HF = 15.8 Hz, 1F). EMAR (EI +) calculated for C 25 H 25 F 2 IN 2 O 3 [M +]: 566.0878, found: 566.0876.

(2R, 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17'a) . 50 mg of 12'a epoxide provided 58 mg (68%) of (17'a) after flash chromatography. [α] D 25 = +18.80 ( c 1.0, CHCl 3).

(2 S , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17'b) . 36 mg of epoxy 12'b led, after chromatography, to 43 mg (70%) of (17'b), [α] D 25 = +15.60 ( c 1.0, CHCl 3 ).

         \ vskip1.000000 \ baselineskip
      

36

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate tert -butyl ester (16a) . To a solution of epoxide 13a (48 mg, 0.16 mmol) in anhydrous isopropanol (1.0 mL) at room temperature 3-iodobenzylamine (0.18 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was purified by flash chromatography, using n- hexane: AcOEt (2: 1) as eluent, 74 mg of amino alcohol 16a (87%) being isolated as a clear oil. [α] D 25 = -17.99 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.34 (s, 9H), 2.58 (dd, J = 12.0, 9.0 Hz, 1H), 2.67 (dd, J = 12.0, 4.5 Hz, 1H), 3.65 (d, J = 13.5 Hz, 1H), 3.72 (d, J = 13.5 Hz, 1H), 4.01 (dd, J = 9.0, 4.5 Hz, 1H), 4.06-4.16 (m, 1H), 5.28 (d, J = 10.2 Hz, 1H), 7.03 (t, J = 7.5 Hz, 1H), 7.19 (d, J = 7.5 Hz, 1H), 7.41-7.62 (m, 7H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 51.4, 52.6, 56.5 (t, J CF = 28.2 Hz), 65.7, 79.9, 94.4, 121.8 (t, J _ {CF = 249.3 Hz), 125.6 (t, J CF = 6.3 Hz), 127.2, 128.2, 130.0, 130.2, 134.8 (t, J CF = 25.8 Hz), 136.2, 136.9, 142.0, 155.5. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.3 (dd, J HF = 14.4 Hz, 1F), -104.3 (dd, J HF = 13.0 Hz, 1F ). EMAR (FAB) calculated for C_22 H_ {28} F_ {2} IN_ {2} [M + H +]: 533.1113, found: 533.1092.

         \ vskip1.000000 \ baselineskip
      

37

(2 R , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate tert -butyl ester (16b) . 21.5 mg of epoxide 13b (0.07 mmol) led to 16b (35.6 mg, 93%). White solid, mp: 100-102 ° C (CH 2 Cl 2). [α] D 25 = -10.25 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.22 (sa, 1H), 1.34 (s, 9H), 2.31 (sa, 1H), 2.75-2.84 (m, 2H), 3.69 ( d, J = 13.8 Hz, 1H), 3.74 (d, J = 13.8 Hz, 1H), 3.89 (dd, J = 9.2, 4.7 Hz, 1H), 4.32-4.49 (m, 1H), 5.46 (d, J = 9.6 Hz, 1H), 7.05 (t, J = 7.5 Hz, 1H), 7.26 (d, J = 7.5 Hz, 1H), 7.38-7.45 (m, 3H), 7.45-7.53 (m, 2H), 7.58 (d, J = 7.9 Hz, 1H), 7.65 (s, 1H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 50.8, 53.1, 58.9 (t, J CF = 27.2 Hz), 67.9, 80.1, 94.4, 121.6 (t, J _ {CF = 248.9 Hz), 125.4 (t, J CF = 6.0 Hz), 127.3, 128.3, 130.0, 130.2, 135.0 (t, J CF = 25.4 Hz), 136.1, 137.0, 142.2, 155.8. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.6 (dd, J FF = 249.6 Hz, J HF = 11.6 Hz, 1F), -101.8 (dd, J FF = 249.6 Hz, J HF = 16.4 Hz, 1F). EMAR (FAB) calculated for C_22 H_ {28} F_ {2} IN_ {2} [M + H +]: 533.1113, found: 533.1113.

(2 S , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate tert- butyl (16'a) . 35 mg of epoxide 13'a led to 53 mg, 89% of 16'a [α] D 25 = +22.57 ( c 1.0, CHCl 3).

(2 S , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate tert- butyl (16'b) . 67 mg of epoxy 13'b led to 74 mg, 62% of 16'b. [α] D 25 = +11.32 ( c 1.0, CHCl 3).

         \ vskip1.000000 \ baselineskip
      

38

N - (4-Bromophenyl) sulfonyl [(1 R ) -1- (phenyldifluoromethyl) allyl] amine (9) . To a solution of allylamine 4 (100 mg, 0.3 mmol) in anhydrous methanol (2.2 mL) was added 0.87 mL of a solution of HCl (4 M) in dioxane, and allowed to stir at room temperature for 2 hours After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 2.2 mL of dioxane. Then, K 2 CO 3 (1.0 mmol), (4-bromophenyl) sulfonyl chloride (1.7 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.05) were added successively at room temperature mmol). The reaction mixture was left with continuous stirring at room temperature for 18 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to flash chromatography, using n- hexane: ethyl ether (6: 1) as eluent, obtaining a white solid corresponding to allylamine 9 (92 mg, 66%) as a white solid, mp: 94-96 ° C (CH 2 Cl 2). [α] D 25 = +4.12 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 4.47 (tddt, J = 11.6, 9.6, 6.0, 2.1 Hz, 1H), 5.13 (d, J = 9.6 Hz, 2H), 5.24 ( d, J = 18.0 Hz, 1H), 5.25 (d, J = 10.2 Hz, 1H), 6.75 (ddd, J = 18.0, 10.2, 6.0 Hz, 1H), 7.40-7.43 (m, 3H), 7.47-7.50 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 61.4 (t, J CF = 30.6 Hz), 120.2 (t, J CF = 249.4 Hz), 120.6, 125.7 ( J CF = 6.2 Hz), 127.5, 128.4, 128.4, 130.1 (t, J CF = 2.9 Hz), 130.3, 132.1, 133.5 (t, J CF = 25.7 Hz), 139.6. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.4 (d, J HF = 11.6 Hz, 2F). HRMS (EI +) calculated for C 16 H 14 BrF 2 NO 2 M [M +]: 400.9897, found: 400.9904.

39

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 S ) - N - {2- [1-Phenyl-1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18a) .

Isomer (2 R , 3 S ) . To a solution of the epoxide (49.8 mg, 0.15 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (3-fluorophenylmethyl) amine (0.34 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 16 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 18a (51.8 mg) as a white solid with a white solid. 76% yield Mp: 77-81 ° C. [α] D 25 = -23.68 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.71 (sa, 2H), 2.53-2.60 (m, 2H), 2.65-2.70 (m, 2H), 3.71 (s, 2H), 3.90 (dd J = 9.6, 4.2 Hz, 1H), 4.10-4.23 (m, 1H), 5.00 (d, J = 12.4 Hz, 2H), 5.55 (d, J = 9.9 Hz, 1H), 6.91-7.01 ( m, 3H), 7.22-7.28 (m, 3H), 7.27-7.28 (m, 1H), 7.31-7.35 (m, 2H), 7.40-7.44 (m, 3H), 7.49-7.51 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 51.3, 52.7, 57.1 (t, J CF = 28.1 Hz), 65.6, 67.0, 114.0 (d, J CF = 21.4 Hz), 114.7 (d, J CF = 21.2 Hz), 121.6 (t, J CF = 248.9 Hz), 123.4 (d, J CF = 2.7 Hz), 125.5 (t, J CF = 6.3 Hz), 127.8, 128.1, 128.4 (d, J CF = 8.9 Hz), 129.9 (d, J CF = 8.2 Hz), 130.7, 134.5 (t, J _ {CF = 25.7 Hz), 136.1, 142.0 (d, J CF = 6.7 Hz), 156.3, 161.2, 164.5. 19 F-NMR (CDCl 3, 282.4 MHz) δ -103.4 (dd, J FF = 249.2 Hz, J HF = 14.2 Hz, 1F), -104.6 (d, J FF = 248.9 Hz, J HF = 13.2 Hz, 1F), -113.6 (tdd, J CF = 24.2, 9.0, 6.0 Hz, 1F). EMAR (ESI +) calculated for C 25 H 25 F 3 N 2 O 3 [M + H +]: 459.1896, found: 459.1899.

         \ vskip1.000000 \ baselineskip
      

40

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (47.5 mg, 0.14 mmol) in anhydrous isopropanol (1.9 mL) was added, at room temperature, the primary amine (3-fluorophenylmethyl) amine (0.32 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 18b (38.7 mg) as a white solid with a white solid. 61% yield. Mp: 134-137 ° C. [α] D 25 = -20.51 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 2.1 (sa, 2H), 2.76-2.88 (m, 2H), 3.74 (d, J = 13.8 Hz, 2H), 3.90 (dd J = 9.9, 5.7 Hz, 1H), 4.43-4.57 (m, 1H), 4.27-4.58 (m, 1H), 5.02 (d, J = 12.3 Hz, 2H), 5.06 (s, 2H), 7.26-7.56 ( m, 10H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 51.0, 53.7, 60.0 (t, J CF = 26.5 Hz), 67.5, 68.1, 114.4 (d, J CF = 21.2 Hz), 115.2 (d, J CF = 21.1 Hz), 121.9 (t, J CF = 251.6 Hz), 123.9 (d, J CF = 2.8 Hz), 125.8 (t, J CF = 6.3 Hz), 128.3, 128.5, 128.8, 128.9, 130.3 (d, J CF = 8.3 Hz), 130.6, 134.2, 135.1 (t, J CF = 26.3 Hz), 136.4, 142.6, 156.9. 19 F-NMR (CDCl 3, 282.4 MHz) δ -101.89 (dd, J FF = 251.0 Hz, J HF = 15.8 Hz, 1F), -104.8 (d, J FF = 250.2 Hz, J HF = 12.1 Hz, 1F), -113.7 (tdd, J CF = 28.4, 8.9, 6.1, 2.8 Hz, 1F). EMAR (FAB) calculated for C 25 H 25 F 3 N 2 O 3 [M + H +]: 458.1817, found: 458.1810.

41

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 S ) - N - {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-difluoro-3-hydroxy]} benzyl carbamate (19a) .

Isomer (2 R , 3 S ) . To a solution of the epoxide (43.1 mg, 0.13 mmol) in anhydrous isopropanol (1.8 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.19 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 8 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (1: 1) to obtain product 19a (45.6 mg) as a white solid with a 74% yield. Mp: 89-92 ° C. [α] D 25 = -24.89 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 2.32 (sa, 2H), 2.56-2.72 (m, 2H), 3.71 (s, 2H), 4.02 (dd, J = 4.2 Hz, 1H), 4.14-4.28 (m, 1H), 5.07 (d, J = 12.3 Hz, 2H), 5.61 (d, J = 9.9 Hz, 1H), 7.11-7.15 (m, 1H), 7.24-7.31 (m , 5H), 7.34-7.38 (m, 3 H), 7.40-7.49 (m, 3 H), 7.52, 7.55 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 51.4, 52.7, 57.2 (t, J CF = 28.5 Hz), 65.6, 67.0, 121.6 (t, J CF = 248.5 Hz), 125.5 (t, J CF = 6.4 Hz), 126.0, 127.3, 127.8, 128.0, 128.1, 128.3, 128.4, 129.7, 130.1, 134.3, 134.6 (t, J CF = 25.7 Hz ), 141.6, 156.3. 19 F-NMR (CDCl 3, 282.4 MHz) δ -103.4 (dd, J FF = 249.0 Hz, J HF = 14.6 Hz, 1F), -104.6 (dd, J FF = 249.0 Hz, J HF = 13.2 Hz, 1F). EMAR (EI +) calculated for C 25 H 25 ClF 2 N 2 O 3 [M +]: 474.1522, found: 474.1518.

         \ vskip1.000000 \ baselineskip
      

42

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) -N- {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-difluoro-3-hydroxy]} benzyl carbamate (19b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (46.4 mg, 0.14 mmol) in anhydrous isopropanol (1.9 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.31 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 19b (39.9 mg) as a white solid with a white solid. 60% yield Mp: 120-123 ° C. [α] D 25 = -17.3 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.60 (sa, 2H), 2.76-2.87 (m, 2H), 3.71 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 9.4, 5.4 Hz, 1H), 4.43-4.57 (m, 1H), 5.06 (d, J = 12.3 Hz, 2H), 5.99 (d, J = 9.9 Hz, 1H), 6.91-7.05 (m, 3H ), 7.21-7.50 (m, 11H). 13 C-NMR (75.5 MHz, CDCl 3) δ 51.0, 53.6, 60.1 (t, J CF = 28.7 Hz), 67.5, 68.0, 121.8 (t, J CF = 251.4 Hz), 125.8 (t, J CF = 6.4 Hz), 126.5, 127.7, 128.3, 128.5, 128.8, 128.9, 130.1, 130.6, 134.6, 135.7 (t, J CF = 25.4 Hz), 136.5, 142.1, 157.0. ^ {19} F-NMR (CDCl 3 {}, 282.4 MHz) \ delta -101.8 (dd, J = 250.3 {FF} Hz, J = 16.0 {HF} Hz, 1F), -104.8 (dd, J FF = 250.4 Hz, J HF = 11.2 Hz, 1F), -113.7. EMAR (EI +) calculated for C 25 H 25 ClF 2 N 2 O 3 [M +]: 474.1522, found: 474.1516.

         \ vskip1.000000 \ baselineskip
      

43

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} benzyl carbamate (20b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (42.4 mg, 0.34 mmol) in anhydrous isopropanol (1.7 mL) was added, at room temperature, the primary amine (3-methoxyphenylmethyl) amine (0.48 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (1: 2) and 1% to obtain product 20b (50.0 mg) as a 84% white solid. of performance. Mp: 101-105 ° C. [α] D 25 = -18.95 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 1.59 (sa, 2H), 2.76-2.90 (m, 2H), 3.71 (d, J = 15.0 Hz, 2H), 3.77 (s, 3H), 3.86-3.91 (m, 1H), 4.41-4.57 (m, 1H), 5.03 (d J = 12.3 Hz, 2H), 5.86 (d, J = 10.2 Hz, 1H), 6.78-6.90 (m, 3H), 7.18-7.24 (m, 2H), 7.32-7.50 (m, 9H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 50.6, 53.7, 55.1 59.7, (t, J CF = 28.5 Hz), 67.0, 67.4, 112.6, 113.4, 121.7 (t, J CF = 248.3 Hz), 120.3, 125.4 (t, J CF = 6.3 Hz), 127.8, 128.0, 128.3, 128.4, 129.4, 130.1, 134.8 (t, J CF = 25.5 Hz ), 136.1, 141.1, 156.5, 159.7. 19 F-NMR (CDCl 3, 282.4 MHz) δ -1041.4 (dd, J FF = 250.7 Hz, J HF = 14.6 Hz, 1F), -104.9 (dd, J FF = 250.7 Hz, J HF = 12.1 Hz, 1F). EMAR (ESI <+>) calculated for C_ {26} H_ {28} F_ {N} {2} {4} [M + H +]: 471.2095, found: 471.2090.

         \ vskip1.000000 \ baselineskip
      

44

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) -N- {2- [1-Phenyl-1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} benzyl carbamate (21b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (49.0 mg, 0.12 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.26 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 21 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) to obtain product 21b (43.3 mg) as a white solid in 83% yield. Mp: 133-136 ° C. [α] D 25 = -20.10 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.60 (sa, 2H), 2.76-2.87 (m, 2H), 3.71 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 9.4, 5.4 Hz, 1H), 4.43-4.57 (m, 1H), 5.06 (d, J = 12.3 Hz, 2H), 5.99 (d, J = 9.9 Hz, 1H), 6.91-7.05 (m, 3H ), 7.21-7.50 (m, 11H). 13 C-NMR (75.5 MHz, CDCl 3) δ 48.2, 50.4, 59.6 (t, J CF = 27.4 Hz), 67.0, 67.6, 121.3, (t, J CF) = 248.4 Hz), 124.5, 125.0, 125.4 (t, J CF = 6.3 Hz), 126.6, 127.8, 128.0, 128.3, 128.4, 130.1, 134.8 (t, J CF = 25.6 Hz), 136.1 , 143.2, 156.5. 19 F-NMR (CDCl 3, 282.4 MHz) δ -101.9 (dd, J FF = 250.4 Hz, J HF = 16.0 Hz, 1F), -104.7 (dd, J FF = 250.3 Hz, J HF = 11.8 Hz, 1F). EMAR (ESI +) calculated for C 23 H 24 F 2 N 2 O 3 S [M + H +]: 447.1554, found: 447.1552.

         \ vskip1.000000 \ baselineskip
      

Four. Five

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-4- (2-furylmethyl) amino-3-hydroxy]} benzyl carbamate (22b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (48.7 mg, 0.15 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (2-furylmethyl) amine (0.33 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using pentane: ethyl ether (4: 6) to obtain product 22b (40.7 mg) as a white solid in 65% yield. Mp: 135-140 ° C. [α] D 25 = -19.16 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CD 2 Cl 2) δ 2.26 (sa, 2H), 2.69-2.76 (m, 1H), 2.80-2.85 (m, 1 H), 3.79- 3.84 (m, 1H), 4.37-4.52 (m, 1H), 4.99 (d, J = 12.4 Hz, 1H), 5.80 (d, J = 9.6 Hz, 1H), 6.23 (d, 3.3 Hz, 1H), 6.31-6.32 (m, 1H), 7.22-7.26 (m, 2H), 7.29-7.33 (m, 4H), 7.39-7.49 (m, 5H). 13 C-NMR (75.5 MHz, CDCl 3) δ 46.6, 51.1, 54.4 (t, J CF = 27.4 Hz), 67.6, 68.2, 107.8, 110.9, 122.2 (t, J _ {CF = 253.0 Hz), 126.2 (t, J CF = 6.4 Hz), 128.5, 128.8, 129.1, 129.2, 130.9, 135.7 (t, J CF = 25.6 Hz), 137.3, 142.6, 154.4, 157.1. 19 F-NMR (CDCl 3, 282.4 MHz) δ -101.3 (dd, J FF = 249.9 Hz, J HF = 16.0 Hz, 1F), -104.2 (dd, J FF = 249.6 Hz, J HF = 11.8 Hz, 1F). EMAR (ESI +) calculated for C 23 H 24 F 2 N 2 O 4 [M + H +]: 431.1782, found: 431.1781.

46

(2 R , 3 S ) - N - {2- [1-Phenyl-1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} tert- butyl carbamate (23a) .

Isomer (2 R , 3 S ) . To a solution of the epoxide (50 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (3-fluorophenylmethyl) amine (0.38 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 6 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) to obtain product 23a (53.2 mg) as a yellow oil with a 75% yield. [α] D 25 = -21.9 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.33 (s, 9H), 2.55 (sa, 2H), 2.64-2.69 (m, 2H), 3.73 (d, J = 13.5 Hz, 2H), 4.01-4.05 (m, 1H), 4.06-4.16 (m, 1H), 5.33 (d, J = 9.9 Hz, 1H), 6.90-7.02 (m, 3H), 7.21-7.29 (m, 1H) , 7.38-7.42 (m, 3H), 7.47-7.52 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.5, 51.8, 53.2, 56.9 (t, J CF = 27.9 Hz), 66.1, 80.3, 114.4 (d, J CF = 21.1 Hz), 115.2 (d, J CF = 21.2 Hz), 122.4 (t, J CF = 248.8 Hz), 123.9 (d, J CF = 2.7 Hz), 126.0 ( t, J CF = 6.3 Hz), 128.6, 130.34 (d, J CF = 8.4 Hz), 135.2 (t, J CF = 25.7 Hz), 142.4 (d, J CF } = 6.8 Hz), 155.9, 161.6, 164.9. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.3 (dd, J HF = 19.7, 5.6 Hz, 2 F), -113.6 (td, J CF = 24.5 , 9.2, 6.0 Hz, 1 F). EMAR (ESI <+>) calculated for C_ {22} H_ {27} F_ {3} {2} {3} [M + H +]: 425.2052, found: 425.2053.

47

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} tert- butyl carbamate (23b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (53.2 mg, 0.18 mmol) in anhydrous isopropanol (2.2 mL) was added, at room temperature, the primary amine (3-fluorophenylmethyl) amine (0.67 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) to obtain product 23b (48.9 mg) as a white solid in 65% yield. Mp: 101-104 ° C. [α] D 25 = -12.34 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.33 (s, 9H), 2.48 (sa, 2H), 2.82 (m, 2H), 3.76 (d, J = 13.5 Hz, 2H) , 3.89 (dd, J = 9.6, 5.1 Hz, 1H), 4.32-4.47 (m, 1H), 5.59 (d, J = 9.9 Hz, 1H), 6.91-7.07 (m, 4H), 7.23-7.28 (m , 1H), 7.40-7.50 (m, 4H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 50.8, 53.3, 58.9 (t, J CF = 26.9 Hz), 67.9, 80.1, 113.0 (d, J CF = 21.2 Hz), 114.8 (d, J CF = 21.2 Hz), 121.6 (t, J CF = 251.5 Hz), 123.5 (d, J CF = 2.7 Hz), 125.4 ( t, J CF = 6.3 Hz), 128.3, 130.3 (d, J CF = 8.1 Hz), 135.0 (t, J CF = 25.7 Hz), 142.4 (d, J CF } = 6.7 Hz), 155.7, 161.3, 164.5. 19 F-NMR (CDCl 3, 282.4 MHz) δ -102.7 (dd, J HF = 249.5, 16.3 Hz, 1F), -104.2 (dd, J HF = 249.3, 11.5 Hz, 1F), -113.7 (td, J CF = 28.2, 8.9, 6.1 Hz, 1F). EMAR (FAB) calculated for C_ {22} H_ {27} F_ {3} N2 O_ {3} [M + H +]: 425.2052, found: 425.2047.

48

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 S ) - N - {2- [1-Phenyl-1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert -butyl carbamate (24a) .

Isomer (2 R , 3 S ) . To a solution of the epoxide (50.0 mg, 0.78 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.38 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 24a (45.0 mg) as a yellow oil with a 65% yield. [α] D 25 = -20.90 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.34 (s, 9H), 2.52 (sa, 2H), 2.68-2.74 (m, 2), 3.95 (d, J = 14.4 Hz, 2H), 3.99-4.15 (m, 2H), 5.33 (d, J = 9.9 Hz, 1H), 6.86-6.93 (m, 2H), 7.19 (dd, J = 4.9, 1.2 Hz, 1H), 7.40-7.52 (m, 5H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 47.7, 51.1, 56.5 (t, J CF = 27.9 Hz), 65.6, 79.8, 121.6 (t, J CF = 251.9 Hz), 124.6, 125.1, 125.6 (t, J CF = 6.4 Hz), 126.6, 128.2, 129.9, 134.8 (t, J CF = 25.7 Hz), 143.0, 155.4. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.3 (d, J HF = 14.2 Hz, 2F). EMAR (FAB) calculated for C 20 H 26 C 2 N 2 O 3 S [M + H +]: 413.1710, found: 413.1715.

         \ vskip1.000000 \ baselineskip
      

49

         \ vskip1.000000 \ baselineskip
      

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert -butyl carbamate (24b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (50.8 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.51 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 24b (48.5 mg) as a 69% white solid. of performance. Mp: 95-97 ° C. [α] D 25 = -17.26 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.21 (sa, 2 H), 1.33 (s, 9H), 2.79-2.87 (m, 2H), 3.85-3.90 (m, 1H) , 3.97, (d, J = 14.4 Hz, 2H), 4.32-4.47 (m, 1H), 5.37 (d, J = 9.9 Hz, 1H), 6.90-6.95 (m, 2H), 7.21 (dd, J = 6.3, 1.2 Hz, 1H), 7.40-7.42 (m, 4 H), 7.48-7.5 (m, 2 H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 48.2, 50.5, 55.7, 58.7 (t, J CF = 28.4 Hz), 68.0, 80.1, 121.6 (t, J _ {CF = 251.1 Hz), 124.4, 124.9, 125.4 (t, J CF = 6.4 Hz), 126.6, 128.3, 130.0, 135.0 (t, J CF = 25.4 Hz), 143.4, 155.6. 19 F-NMR (CDCl 3, 282.4 MHz) δ -102.2 (dd, J HF = 247.9, 16.3 Hz, 1F), -104.5 (dd, J HF = 249.9, 11.8 Hz, 1F). EMAR (FAB) calculated for C 20 H 26 C 2 N 2 O 3 S [M + H +]: 412.1632, found: 412.1623.

fifty

(2 R , 3 R ) - N - {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-difluoro-3-hydroxy]} tert- butyl carbamate (25b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (50.1 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.63 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) to obtain product 25b (42.9 mg) as a white solid in 58% yield. Mp: 110-113 ° C. [α] D 25 = -12.34 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.33 (s, 9H), 2.40 (sa, 2H), 2.81 (m, 2H), 3.74 (d, J = 13.6 Hz, 2H) , 3.89 (d, J = 9.9, 5.2 Hz, 1H), 4.32-4.46 (m, 1H), 5.51 (d, J = 9.6 Hz, 1H), 7.16-7.19 (m, 1H), 7.23-7.25 (m , 2H), 7.40-7.42 (m, 3H), 7.47-7.50 (m, 2H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 50.8, 53.3, 58.9 (t, J CF = 26.5 Hz), 68.0, 80.1, 121.6 (t, J CF = 248.5 Hz), 125.4 (t, J CF = 6.2 Hz), 126.1, 127.2, 128.1, 128.3, 129.6, 130.0, 134.2, 135.0 (t, J CF = 25.4 Hz), 141.8, 155.7. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.8 (dd, J HF = 250.2, 11.5 Hz, 1F), -105.5 (dd, J HF = 250.2, 14.6 Hz, 1F). HRMS (FAB) calculated for C 22 H 27 FClN 2 O 3 [M + H +]: 440.1678, found: 439.0976.

51

(2 R , 3 R ) - N - {2- [1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} tert -butyl carbamate (26b) .

Isomer (2 R , 3 R ) . To a solution of the epoxide (54.9 mg, 0.18 mmol) in anhydrous isopropanol (2.3 mL) was added, at room temperature, the primary amine (3-methoxyphenylmethyl) amine (0.55 mmol) and molecular sieve (3 \ ring {A}). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by flash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 26b (17.1 mg) as a 21% yellow oil of performance. [α] D 25 = -12.34 ( c 1.0, CHCl 3). 1 H-NMR (300 MHz, CDCl 3) δ 1.33 (s, 9H), 2.21 (sa, 2H), 2.82 (m, 2H), 3.74 (d, J = 13.5 Hz, 2H) , 3.83 (s, 3H), 3.88 (dd, J = 9.6, 5.4 Hz, 1H), 4.32-4.47 (m, 1H), 5.39 (d, J = 9.9 Hz, 1H), 6.78-6.89 (m, 3H ), 7.20-7.26 (m, 1 H), 7.40-7.51 (m, 5 H). 13 C-NMR (75.5 MHz, CDCl 3) δ 28.1, 50.7, 53.7, 55.1, 58.7 (t, J CF = 26.7 Hz), 67.9, 80.0, 112.4, 113.5, 121.5 ( t, J CF = 249.9 Hz), 120.2, 125.4 (t, J CF = 6.3 Hz), 128.2, 129.4, 130.0, 135.0 (t, J CF = 26.1 Hz), 141.3, 155.6, 159.7. 19 F-NMR (CDCl 3, 282.4 MHz) δ -102.3 (dd, J HF = 254.1, 16.0 Hz, 1F), -104.4 (dd, J HF = 254.1, 11.2 Hz, 1F). HRMS (FAB) calculated for C 23 H 30 F 2 N 2 O 4 [M + H +]: 436.2174, found 436.2127.

52

(2 R , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (dipropylamino) butaneyl-2-carbamate benzyl (27a) . To a solution of epoxide 12a (68 mg, 0.20 mmol) in anhydrous isopropanol (0.7 mL) at room temperature dipropylamine (0.3 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was purified by flash chromatography, using n- hexane: AcOEt (2: 1) as eluent, the product 27a (72 mg, 81%) being isolated as a yellowish solid. Mp = 69-71 ° C (CH 2 Cl 2). [α] D 25 = -51.35 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 0.83 (t, J = 7.3 Hz, 6H), 1.24-1.50 (m, 4H), 2.29-2.46 (m, 4H), 2.29- 2.46 (m, 2H), 3.90 (sa, 1H), 3.92-4.00 (m, 1H), 4.00-4.11 (m, 1H), 5.02 (d, J = 12.3 Hz, 1H), 5.07 (d, J = 12.3 Hz, 1H) 5.58 (d, J = 10.1 Hz, 1H), 7.25.7.46 (m, 8H), 7.50-7.54 (m, 2H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 11.6, 20.1, 55.8, 56.9, 57.1 (t, J CF = 29.6 Hz), 62.9, 66.9, 121.5 (t, J _ {CF = 249.0 Hz), 125.8 (t, J CF = 6.4 Hz), 127.8, 128.0, 128.2, 128.4, 130.0, 134.8 (t, J CF = 26.0 Hz), 136.3, 156.3. 19 F-NMR (CDCl 3, 282.4 MHz) δ -104.8 (dd, J FF = 2489.9 Hz, J HF = 15.1 Hz, 1F), -103.2 (dd, J FF = 248.9 Hz, J HF = 12.2 Hz, 1F). EMAR (FAB) calculated for C_ {24} H_ {33} F_ {2} N2 O_ {3} [M + H +]: 435.2471, found: 435.2467.

53

(2 R , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (benzylamino) butaneyl-2-carbamate benzyl (28b) . To a solution of epoxide 12b (20 mg, 0.06 mmol) in anhydrous isopropanol (0.5 mL) at room temperature, benzylamine (0.15 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was purified by flash chromatography, using n- hexane: AcOEt (2: 1) as eluent, the product 28b (17.2 mg, 65%) being isolated as a white solid. . Mp = 132-134 ° C (CH 2 Cl 2). [α] D 25 = -11.51 ( c 1.0, CHCl 3). 1 H-NMR (CDCl 3, 300 MHz) δ 2.14 (sa, 2H), 2.80 (dd, J = 12.6, 6.2 Hz, 1H), 2.88 (dd, J = 12.6, 3.7 Hz, 1H), 3.72 (d, J = 13.3Hz, 1H), 3.77 (d, J = 13.3Hz, 1H), 3.89 (td, J = 6.2, 3.7 Hz, 1H), 4.43-4.58 (m, 1H), 5.00 (d, J = 12.2 Hz, 1H), 5.06 (d, J = 12.2 Hz, 1H), 6.00 (d, J = 9.9 Hz, 1H), 7.24-7.50 (m, 15H). 13 C-NMR (CDCl 3, 75.5 MHz) δ 50.7, 53.9, 59.7 (t, J CF = 27.0 Hz), 67.1, 67.7, 121.4 (t, J CF = 246.0 Hz), 125.4 (t, J CF = 6.4 Hz), 127.1, 127.9, 128.0, 128.1, 128.4, 128.5, 128.5, 130.2, 134.9 (t, J CF = 25.5 Hz), 136.1, 139.6, 156.5. ^ {19} F-NMR (CDCl 3 {}, 282.4 MHz) \ delta -104.9 (d, J = 250.5 {FF} Hz, 1F), -101.9 (dd, J = 250.5 {FF} Hz, 1F ). EMAR (ESI +) calculated for C 25 H 29 N 2 O 3 [M + H +]: 441.2002, found: 441.1989.

         \ vskip1.000000 \ baselineskip
      

Evaluation of the antimicrobial activity of ethanolamines difluorobenzyl

An initial evaluation was made by studying the antimicrobial activity of the selected synthetic intermediates. Initially all the compounds in Schemes 8, 9 and 10 as well as all those included in Table 1 were subjected to different tests to determine their possible application as antimicrobials, against different species of bacteria, gram positive and gram negative, and against fungi, using agar diffusion technique. This is a semiquantitative technique that allows to determine the inhibition of microbial growth produced by the compound under study.

         \ vskip1.000000 \ baselineskip
      

Agar diffusion method

The in vitro antimicrobial activity of the compounds was determined against various species ( S. aureus ATCC 29213, E. coli ATCC 25922, M. luteus ATCC 49732, M. smegmatis ATCC 19420, and C. albicans ATCC 14053) using a suspension of Frozen cells (0.5 McFarland units) of these microorganisms in Mueller-Hinton culture broth (BBL, BD, Sparks, Md.) with 20% glycerol. After defrosting and homogenizing the previous suspensions, a moistened swab was used to perform a grass seeding on Mueller-Hinton agar plates (BBL, BD, Sparks, Md.). 6 mm sterile paper discs, BBL, BD, Sparks, Md. Were impregnated with the dissolution of antimicrobial agents in DMSO and placed on the surface of the agar. The plates were incubated in the air at 37 ° C overnight before reading the inhibition zones.

A similar methodology was used for the evaluation of activities against M. kansasii, N. asteroides , and N. farcinica . In this case, the suspension used was prepared at 3 McFarland units for the first and 1 McFarland units for Nocardia species. In addition, the medium used for the tests with M. kansasii was Middlebrook and Cohn 7H10 agar (BBL, BD, Sparks, Md.). Plates were incubated at 37 ° C for 7-8 days ( M. kansasii ) or 2-3 days ( Nocardia spp .) Prior to reading the zone of inhibition.

         \ vskip1.000000 \ baselineskip
      

Method of microdilution in broth for Mycobacterium

To polystyrene microtiter plates, 96 wells with round bottom (Corning Inc., Corning, NY) are given added 50 µL of modified 7H10 broth per well. The antimicrobial agents were prepared at a 4X concentration the maximum concentration tested, and 50 µL of this was added dissolution to the first well. They were subsequently performed serial double dilutions, leaving the last well as control positive, without antimicrobial agent. To prepare the insulations bacterial, frozen cultures were thawed and diluted to a final concentration of 1.25 x 10 5 CFU / mL (inoculum of work) in modified 7H10 broth (7H10 agar formulation in the which omitted the agar and malachite green). The final inoculum is measured by titration and seeding on 7H10 agar with 10% OADC; the agar plates were incubated for 1 week at 37 ° C. To each well 50 µL of the work inoculum was added. Plates microtiter were covered with sealing adhesive film SealPlate (Excel Scientific, Wrightwood, CA) and incubated at 37 ° C for 7-8 days prior to reading. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of antimicrobial agent that did not provide turbidity visible. Each bacterial isolate was tested in duplicate.

         \ vskip1.000000 \ baselineskip
      

Broth microdilution method for Nocardia

To polystyrene microtiter plates, 96 wells with round bottom (Corning Inc., Corning, NY) are given added 50 µL of Mueller-Hinton broth modified per well. Antimicrobial agents were prepared to a 4X concentration the maximum concentration tested, and they added 50 µL of this solution to the first well. Subsequently double serial dilutions were made, leaving the last well as a positive control, without antimicrobial agent. To prepare bacterial isolates, frozen cultures thawed and diluted to a final concentration of 1.25 x 10 5 CFU / mL (working inoculum) in broth Mueller-Hinton with cationic supplement (following the procedure approved by the Clinical and Laboratory Standards Insitute, CLSI). The final inoculum was measured by titration and seeding in Mueller-Hinton agar. Agar plates are incubated for 4 days at 37 ° C. 50 wells were added to each well µL of the work inoculum. The microtiter plates are covered with SealPlate sealing adhesive film (Excel Scientific, Wrightwood, CA) and incubated at 37 ° C for 3-4 days before reading. Concentration Minimum inhibitory (MIC) was defined as the lowest concentration of antimicrobial agent that did not provide visible turbidity. Every Bacterial isolate was assayed in duplicate.

The intermediate difluorobenzyl derivatives of the allylamine and epoxide type studied (4 and 12b, respectively) showed no evidence of inhibition of microbial growth while the difluorobenzyl derivative with hydroxyethylamine skeleton 17b showed an antimicrobial activity that proved to be selective against some of the microorganisms tested . Specifically, this compound produced growth inhibition of strains of Mycobacterium smegmatis and Micrococcus luteus but not of other bacteria such as Staphylococcus aureus , Escherichia coli or fungi such as Candida albicans (Table 2, entries 1-5). This selectivity is of special interest since it suggests a specific mechanism of action on special characteristics of some species, as opposed to a nonspecific and generalized mechanism that could be potentially toxic.

         \ vskip1.000000 \ baselineskip
      

54

         \ vskip1.000000 \ baselineskip
      

The bacteria of the Mycobacterium genus constitute a group of special relevance due to their pathogenicity (they are the cause of tuberculosis, leprosy and other pulmonary, cerebral and cutaneous infections among others) and because of the difficulty in their treatment, due to the differential characteristics of Their bacterial wall gives them high resistance to conventional treatments. As a consequence, the search for new active drugs continues to be studied. Therefore, the in vitro evaluation was extended to other Mycobacterium species. It was also extended to Nocardia species, a genus of pathogenic bacteria closely related to Mycobacterium . The results obtained are shown in Table 2, expressed in millimeters, corresponding to the diameter of the growth inhibition halo produced around the disk impregnated in the compound to be tested.

         \ vskip1.000000 \ baselineskip
      

         \ vskip1.000000 \ baselineskip
      

         \ vskip1.000000 \ baselineskip
      

(Table goes to page next)

TABLE 2

55

         \ vskip1.000000 \ baselineskip
      

Compound 17b demonstrated activity against various Mycobacterium species, especially M. smegmatis and M. kansasii . It also produced growth inhibition of N. asteroids and N. farcinica .

         \ vskip1.000000 \ baselineskip
      

Evaluation of in vitro antibacterial activity on Mycobacterium and Nocardia species

Due to the interest of the results obtained in The initial evaluation was decided to deepen the study of others structurally related molecules. So, a small library of products to be tested, in order to carry out a correlation study structure-activity (QSAR). He addressed the Preparation of the rest of diastereoisomers of compound 17b initially studied and also from other molecules with modifications in various functional groups.

Ethambutol (EMB) was used as the reference compound in the tests. An ethambutol analogue, SQ-109 developed by Sequella Inc., which is under investigation due to the good results provided as a potential antituberculous agent, was also included in the study in previous studies ( Journal of Antimicrobial Chemotherapy 2006 , 58 , 332-337, Antimicrobial Agents and Chemotheraphy , 2007 , 1563-1565). Table 3 shows the results of diffusion in agar, in millimeters of inhibition, of the four diastereoisomers of the structure that had provided the best results initially, together with EMB and SQ-109.

56

         \ vskip1.000000 \ baselineskip
      

TABLE 3

57

         \ vskip1.000000 \ baselineskip
      

Agar diffusion assays revealed that the four diastereoisomers were active on various strains of Mycobacteria and Nocardia , especially on N. asteroids 720 , strain on which EMB was inactive, which is especially interesting. Few inhibition differences were observed between diastereoisomers, with 17b antibacterial activity slightly better. EMB proved to be much more active on Mycobacterium kansasii than the new compounds, while in the case of Nocardia the differences were insignificant. SQ-109 provided the best results on both Mycobacterium and Nocardia .

58

         \ vskip1.000000 \ baselineskip
      

Other structural modifications that are performed on the main molecule (compounds, 16a, 16b, 16'a, 16'b, 27a and 28b) following the same process of obtaining that had carried out with 17 but using commercial amines adequate, led to compounds that turned out to be inactive, made which allowed to deduce that both the Cbz cluster on the atom of nitrogen as the presence of the iodine atom on the ring Aromatic are of special importance in biological activity. Preliminary tests of other derivatives with different substitutions aromatic on the amine, subsequently prepared, (compounds 18a, 19a, 18b, 19b, 20b, 21b, 22b, 23a, 24a, 25b, 23b, 26b and 24b), obtained similarly to the previous ones seemed to suggest that the most favorable combination for antimicrobial activity was the of product 17b, resulting in the rest of the less active analogs than this one or even completely inactive.

In view of the results, we proceeded to analyze the data with a technique that allowed to determine the bacteriostatic activity quantitatively and in liquid medium, since sometimes the agar diffusion technique can give false negatives or inaccurate data due to problems of diffusion of molecules in agar. Therefore the method was used of determination of the minimum inhibitory concentration (MIC) by microdilution in culture broth. CMI data (expressed in µg / ml) obtained for the four diastereoisomers together with the of EMB and SQ-109 are shown in Table 4.

         \ vskip1.000000 \ baselineskip
      

TABLE 4

59

It should be noted that while on Mycobacterium kansasii the MIC of EMB is much lower than that of the new derivatives and even better than that of SQ-109, when it comes to Nocardia, the ethanolamine type derivatives are more potent than EMB (this is relatively inactive ), presenting lower CMI, and very similar in activity to SQ-109, being in this case 17b and 17'b almost as active as SQ-109, with slightly significant differences in CMI. Therefore, it can be concluded that fluorinated ethanolamine compounds (especially 17b and 17'b) are more active against Nocardia than EMB with MICs similar to those of SQ-109.

Difluorobenzyl ethanolamines according with the invention they can be used in the preparation of a medication for the treatment of leprosy, tuberculosis or some conditions related to any of these diseases. For this, you can make a composition pharmaceutical characterized in that it comprises at least one stereoisomer of one or more difluorobenzyl ethanolamines or one of its pharmaceutically acceptable salts, together with one or more vehicles and / or diluents necessary to be administered via oral, transdermal, parental or inhalative. In such cases, the Difluorobenzyl ethanolamines are presented as constituents assets in the appropriate forms of presentation for the various forms of administration such as tablets, capsules, suppositories, solutions, juices, emulsions or dispersible powders. The tablets may also be delayed release.

         \ vskip1.000000 \ baselineskip
      

References

1 Spigetman MK, The Journal of Infectious Diseases , 2007 , 196, S28-34.

2 Sacchettini , Jc Rubin , EJ, Freudlinch , JS, Microbiology 2008 , 6, 41-52.

3 Chatterjee , D., Biopolymers , 1997 , 1, 579-588.

4 Gutierrez-Lugo , MT, Bewley , CA, J. Med. Chem . 2008 , 51, 2606-2612.

5 Ralp , AP, Anstey , NM, Kelly , PM, Clinical Infectious Diseases , 2009 , 49, 574-583.

6 Kuduk , SD, Marco , CND, Pitzenberger , SM, Tsou , N., Tetranhedron Lett ., 2006 , 47, 2377-2381.

7 Philippe , Christine et al . "Synthesis of New Triifluoromethylated Hydroxyethylamine-Based Scaffolds" Eur. J. Org. Chem 2009 , 5215-5223.

8 Dos Santos , Mickael et al ., Tetrahedron Letters , 2009 , 50, 857-859.

9 Purser , S., Moore , PR; Swallow , S .; Governeur , V., Chem. Soc. Rev. 2008 , 37, 320-330; Muller , K .; Faeh , C .; Diederich , F., Science , 2007 , 317, 1881-1886; Hagmann , WK, J. Med. Chem . 2008 , 51, 4359-4369.

10 Khetan , SK; Collins , TJ Chem. Rev. 2007 , 107, 2319-2364.

11 Scott E. Denmark et al ., J. Org. Chem 1995 , 60, 1391-1407.

12 R. Smits , CD Cadicamo , K. Burger , B. Koksch , Chemical Society Reviews 2008 , 37, 1727-1739.

13 S. Fustero , JF Sanz-Cerverea , JL Aceña , M. Sánchez-Roselló , Synlett 2009 , 525-549.

14 Sani , M .; Volontery , A .; Zanda , M., ChemMedChem . 2007 , 2, 1693-1700; Pesenti , C .; Arnone , A .; Bellosta , S .; Bravo , P .; Canavesi , M .; Corradi , E .; Frigerio , M .; Meille , SV; Monetti , M .; Panzeri , W.; Viani. F.; Venturini , R .; Zanda , M., Tetrahendron , 2001 , 57, 6511-6522.

15 Fustero , S .; Flores , S .; Cuñat , AC; Jiménez , D .; From the Well ; C., Sanz-Cervera , JF, Journal of Fluorine Chemistry 2007 , 128, 1248-1254.

16 Liu , G .; Cogan , DA; Owens , TD; Tang TP, Ellman , JA, J. Org. Chem 1999 , 64, 1278-1284.

17 Troung , VL; Menard , MS; Dion , I., Org. Lett . 2007 , 9, 683-685.

18 Although a representative sample was conveniently separated for the characterization of both isomers

19 Peltier , HM; McMahon , JP; Patterson , AW; Ellman , JA J. Am. Chem. Soc . 2006 , 128, 16018-16019.

20 Fawcett , J .; Griffith , G .; Percy , JM; Uneyama , E. Org. Lett . 2004 , 6, 1277-1280.

Claims (22)

1. Difluorobenzyl ethanolamines represented by the general formula (I), where R is a protective group and Ar is a derived from benzene, furan, thiophene or pyridine.

           \ vskip1.000000 \ baselineskip
        

60

           \ vskip1.000000 \ baselineskip
        

Where: a) the protecting group R may be benzyloxycarbonyl, tert- butyloxycarbonyl; Y b) Ar are derivatives of benzene, furan, thiophene or pyridine, which they can independently possess as Substituents fluorine, chlorine, bromine, iodine, methoxy.

           \ vskip1.000000 \ baselineskip
        

2. Any difluorobenzyl ethanolamine (I) according to claim 1, characterized in that it is composed of a mixture of different isomers. 3. Any difluorobenzyl ethanolamine (I) according to claim 1, characterized in that it is composed of a single isomer. 4. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert - butyl carbamate and Ar is 3-iodophenyl and is selected from (2 R , 3 S ) -1-Phenyl-1,1 difluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate (16a); 16b: (2 R, 3 R) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate (16b):

           \ vskip1.000000 \ baselineskip
        

61

           \ vskip1.000000 \ baselineskip
        

(2 S , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate tert - butyl (16'a) or (2 S , 3 S ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate (16'b):

           \ vskip1.000000 \ baselineskip
        

62

           \ newpage
        

5. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-iodophenyl and is selected from (2 R , 3 S ) -1-Phenyl-1,1-difluoro Benzyl-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate (17a); (2 R , 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17b):

           \ vskip1.000000 \ baselineskip
        

63

           \ vskip1.000000 \ baselineskip
        

(2S, 3 R ) -1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate of benzyl (17'a) or (2 S , 3 S ) -1 -Phenyl-1,1-difluoro-3-hydroxy-4- (3-iodobenzylamino) benzyl-butane-2-carbamate (17'b).

           \ vskip1.000000 \ baselineskip
        

64

           \ vskip1.000000 \ baselineskip
        

6. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-fluorophenyl and is selected from (2 R , 3 S ) - N - {2- [1-Phenyl -1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18a) or (18b): (2 R , 3 R ) - N - {2- [1-Phenyl- Benzyl 1,1-difluoro-4- (3-fluoro-phenylmethyl) amino-3-hydroxy] carbamate (18b): 65

           \ vskip1.000000 \ baselineskip
        

7. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-chlorophenyl and is selected from (2 R , 3 S ) - N - {2- [4- ( 3-Chlorophenylmethyl) amino-1-phenyl-1,1-difluoro-3-hydroxy]} benzyl carbamate (19a) or (2 R , 3 R ) - N - {2- [4 (3-Chlorophenylmethyl) amino- Benzyl 1-phenyl-1,1-difluoro-3-hydroxy]} carbamate (19b): 66 8. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-methoxyphenyl, such as (2 R , 3 R ) - N - {2- [1- Phenyl-1,1-difluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} benzyl carbamate (20b):

           \ vskip1.000000 \ baselineskip
        

67 9. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 2-thienyl, such as (2 R , 3 R ) -N- {2- [1- Phenyl-1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} benzyl carbamate (21b). 68

           \ vskip1.000000 \ baselineskip
        

10. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 2-furyl, such as (2 R , 3 R ) - N - {2- [1- Phenyl-1,1-difluoro-4- (2-furylmethyl) amino-3-hydroxy]} benzyl carbamate (22b): 69

           \ vskip1.000000 \ baselineskip
        

11. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert - butyl carbamate and Ar is 3-fluorophenyl, as for example (2 R , 3 S ) - N - {2- [1 -Phenyl-1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} tert -butyl carbamate (23a) or (2 R , 3 R ) - N - {2- [1-Phenyl -1,1-difluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} tert - butyl carbamate (23b). 70

           \ vskip1.000000 \ baselineskip
        

12. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protective group is tert - butyl carbamate and Ar is 2-thienyl, such as (2 R , 3 S ) - N - {2- [ 1-Phenyl-1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert - butyl carbamate (24a) or (2 R , 3 R ) - N - {2- [1-Phenyl -1,1-difluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert - butyl carbamate (24b):

           \ vskip1.000000 \ baselineskip
        

71

           \ vskip1.000000 \ baselineskip
        

13. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert - butyl carbamate and Ar is 3-chlorophenyl, such as (2 R , 3 R ) - N - {2- [ 4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-difluoro-3-hydroxy]} tert - butyl carbamate (25b):

           \ vskip1.000000 \ baselineskip
        

72

           \ vskip1.000000 \ baselineskip
        

14. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert - butyl carbamate and Ar is 3-methoxyphenyl, such as (2 R , 3 R ) - N - {2- [ 1-Phenyl-1,1-difluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} tert - butyl carbamate (26b):

           \ vskip1.000000 \ baselineskip
        

73

           \ vskip1.000000 \ baselineskip
        

15. Difluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is phenyl, such as (2 R , 3 R) 1-Phenyl-1,1-difluoro-3 benzylhydroxy-4- (benzylamino) butanyl-2-carbamate (28b)

           \ vskip1.000000 \ baselineskip
        

74

           \ vskip1.000000 \ baselineskip
        

16. Process for obtaining difluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof according to claims 1-15, characterized in that it is obtained from difluorobenzyl allylamines 4, or any of its isomers, by the following steps : a) Introduction of the protective group R by reaction of difluorobenzyl allylamine 4 with the compound that contains the desired protecting group, for example with benzyloxycarbonyl to obtain carbamate 10: 75 b) Epoxides formation intermediates 12a and 12b by reaction of allylamines difluorobenzyl 10 for example with trifluoromethyldioxyran: 76 c) Reaction of epoxides 12a and 12b with the desired compound containing the desired Ar group introduce, for example, with 3-iodobenzylamine to form difluorobenzyl ethanolamine 17a: 77 17. Procedure for obtaining the difluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof, according to claim 16, characterized in that the isomers of the epoxides obtained in step (b), are separated, carrying out the reaction of step (c) with each isomer independently. 18. Procedure for obtaining the difluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof, according to claim 16, characterized in that the isomers of the epoxides obtained in step (b) are separated by column chromatography with silica gel 19. Use of ethanolamines difluorobenzyl (I) or a pharmaceutically salt thereof acceptable, according to the claims 1-15, for the preparation of a medicine for antimicrobial treatment 20. Use of an ethanolamine difluorobenzyl (I) or one of its isomers or one of its salts Pharmaceutically acceptable, according to claim 19, for the preparation of a medicine for the treatment of tuberculosis. 21. Use of an ethanolamine difluorobenzyl (I) or one of its isomers or one of its salts Pharmaceutically acceptable, according to claims 19 and 20, for the preparation of a medicament for the treatment of leprosy. 22. Pharmaceutical composition according to claims 1 to 15, characterized in that it comprises at least one stereoisomer of one or more difluorobenzyl ethanolamines or one of its pharmaceutically acceptable salts, according to claims 1 to 19, together with one or more vehicles and / or diluents necessary to be used according to claims 19-21.