Organoselenium compounds, method for producing same, and pharmaceutical uses thereof in particular as antitumor agents

Abstract

The invention relates to a selenium compound. Said selenium compound has formula (I), where R.sup.1=alkyl; R.sup.2=H, R.sup.4C(=0), R.sup.4OC(=0), a-aminoacyl, CH.sub.3SeCH.sub.2CH.sub.2CH(NH.sub.2)C(=0), CH.sub.3SeCH.sub.2CH.sub.2CH(OH)C(=0); X=OH, OR.sup.3, NH.sub.2, NR.sup.4R.sup.5, -amino acid, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)NH, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)0-; R.sup.3=alkyl; R.sup.4=alkyl, aryl; R.sup.5=H, alkyl, aryl; R.sup.4 and R.sup.5 which can together form a 5- or 6-membered cycloalkyl radical which can comprise a heteroatom; provided that when X=NH-terbutyl, R.sup.2C(=0)CH.sub.3. Said compound can be used as a pharmaceutical substance, in particular as an antitumour substance.

Claims

1. A selenium compound having the following general formula (I): ##STR00143## where R.sup.1=a radical selected from a non substituted linear, branched or cyclic saturated carbon containing radical with 1 to 26 carbon atoms, and a substituted linear, branched or cyclic saturated carbon containing radical with 1 to 26 carbon atoms which is substituted by at least one substituent selected from the group consisting of one or more fluorine atoms, one or more carbon-carbon double bonds, at least one OH group, and at least one amino group; R.sup.2=H, R.sup.4C(O), R.sup.4OC(O), -aminoacyl, CH.sub.3SeCH.sub.2CH.sub.2CH(NH.sub.2)C(O), CH.sub.3SeCH.sub.2CH.sub.2CH(OH)C(O); X=OH, OR.sup.3, NH.sub.2, NR.sup.4R.sup.5, -amino acid, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)NH, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)O; R.sup.3=alkyl; R.sup.4=alkyl, aryl; R.sup.5=H, alkyl, aryl; R.sup.4 and R.sup.5 being capable of forming together a 5- or 6-membered cycloalkyl radical which can comprise a heteroatom; provided that XNH-tert-butyl, and any stereoisomers or the pharmaceutically accepted acid or base salts thereof.

2. The selenium compound according to claim 1, wherein R.sup.1 represents a methyl, ethyl, or allyl group.

3. The selenium compound according to claim 1, wherein R.sup.2 is selected from the group consisting of H, -aminoacyls, R.sup.4(CO), R.sup.4O(CO), and CH.sub.3SeCH.sub.2CH.sub.2CH(OH)C(O).

4. The selenium compound according to claim 1, wherein X is selected from the group consisting OH, -amino acid, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)NH, and CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)O.

5. The selenium compound according to claim 1, wherein R.sup.1 represents a methyl, ethyl, or allyl group; R.sup.2 represents R.sup.4(CO), or R.sup.4O(CO), and X represents OH or OR.sup.3.

6. The selenium compound according to claim 1, wherein the pharmaceutically acceptable acids are selected from mineral acids comprising hydrochloric, hydrobromic, hydroiodic, sulfuric, and phosphoric acids, or selected from organic acids comprising formic, acetic, trifluoroacetic, propionic, tartaric, benzoic, maleic, fumaric, succinic, citric, oxalic, glyoxylic, and aspartic acids, alkanesulfonic acids comprising methanesulfonic, trifluoromethanesulfonic, and ethanesulfonic, and arylsulfonic acids comprising benzene- and paratoluenesulfonic acids.

7. The selenium compound according to claim 1, wherein the pharmaceutically acceptable bases are selected from mineral bases comprising sodium, lithium, calcium, potassium, magnesium, ammonium or zinc hydroxides, carbonates of alkali or alkaline earth metals comprising sodium, lithium, calcium, potassium, magnesium, ammonium or zinc carbonates and bicarbonates, or organic bases comprising methylamine, propylamine, trimethylamine, diethylamine, triethylamine, N,N-dimethylethanolamine, tris(hydroxymethyl)aminomethane, ethanolamine, pyridine, picoline, dicyclohexylamine, morpholine, proceine, lysine, arginine, histidine, and N-methylglucamine, or else phosphonium salts comprising the alkyl-phosphonium salts, the aryl-phosphonium salts, the alkyl-aryl-phosphonium salts, and the alkenyl-aryl-phosphonium salts, or quaternary ammonium salts.

8. The selenium compound according to claim 1, wherein said compound is selected from the group consisting of: 2-hydroxy-3-(methylseleno)propanoic acid methyl ester; (R)-2-hydroxy-3-(methylseleno)propanoic acid methyl ester; (S)-2-hydroxy-3-(methylseleno)propanoic acid methyl ester; 2-hydroxy-3-(methylseleno)propanoic acid ethyl ester; (S)-(2-hydroxy-3-(methylseleno)propanoic acid benzyl ester; 3-(ethylseleno)-2-hydroxypropanoic acid methyl ester; 2-hydroxy-3-(isobutylseleno)propanoic acid methyl ester; 2-hydroxy-3-(methylseleno)propanoic acid isopropyl ester; 2-hydroxy-3-(methylseleno)propanoic acid; (R)-2-hydroxy-3-(methylseleno)propanoic acid; (S)-2-hydroxy-3-(methylseleno)propanoic acid; 3-ethylseleno-2-hydroxypropanoic acid; 2-hydroxy-3-(isobutylseleno)propanoic acid; dicyclohexylammonium 2-hydroxy-3-(methylseleno)propanoate salt; sodium 2-hydroxy-3-(methylseleno)propanoate salt; magnesium bis(2-hydroxy-3-(methylseleno)propanoate salt; zinc bis(2-hydroxy-3-(methylseleno)propanoate salt; calcium bis(2-hydroxy-3-(methylseleno)propanoate salt; 2-hydroxy-3-(methylseleno)propanamide; N-cyclopropyl-2-hydroxy-3-(methylseleno)propanamide; N-[2-(dimethylamino)ethyl]-2-hydroxy-3-(methylseleno)propanamide; 2-hydroxy-3-(methylseleno)-1-(pyrrolidin-1-yl)propan-1-one; 2-hydroxy-3-(methylseleno)-1-(piperidin-1-yl)propan-1-one; 2-hydroxy-3-(methylseleno)-1-(morpholin-4-yl)propan-1-one; N, N-diethyl-2-hydroxy-3-(methylseleno)propanamide; 2-hydroxy-N-(2-hydroxyethyl)-3-(methylseleno)propanamide; [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-alanine; [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine methyl ester; [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine; [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine methyl ester; [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine; [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine methyl ester; [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine; N()-[(2RS)-2-hydroxy-3-methylselenopropanoyl]-N()-tert-butoxycarbonyl-(S)-lysine methyl ester; N()-[(2RS)-2-hydroxy-3-methylselenopropanoyl]-N()-fluorenylmethyloxycarbonyl-(S)-lysine methyl ester; N()-[(2 RS)-2-hydroxy-3-methylselenopropanoyl]-N()-benzyloxycarbonyl-(S)-lysine methyl ester; [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine methyl ester; [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine; [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine methyl ester; [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine; [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine methyl ester; [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine; 2-(acetyloxy)-3-(methylseleno)propanoic acid; 2-(dodecanoyloxy)-3-(methylseleno)propanoic acid; 2-(benzoyloxy)-3-(methylseleno)propanoic acid methyl ester; 2-(benzoyloxy)-3-(methylseleno)propanoic acid; 3-(methylseleno)-2-[(3-pyridine)oxycarbonyl]propanoic acid methyl ester; 2-[(tert-butoxycarbonyl)oxy]-3-(methylseleno)propanoic acid methyl ester; 2-[(tert-butoxycarbonyl)oxy]-3-(methylseleno)propanoic acid; (2RS)[N-(tert-butoxycarbonyl)-S-methionyl]-3-(methylseleno)propanoic acid methyl ester; 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid methyl ester; 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid; 3-methylseleno-2-(2-acetoxy-4-methylselenobutanoyl)propanoic acid methyl ester; 3-methylseleno-2-(2-hydroxy-4-methylselenobutanoyl)propanoic acid; 2-(pentanoyloxy)-3-(methylseleno)propanoic acid; 2-(nonanoyloxy)-3-(methylseleno)propanoic acid; 2-(linoleoyloxy)-3-(methylseleno)propanoic acid; 2-(linoleoyloxy)-3-(methylseleno)propanoic acid methyl ester; 2-(pivaloyloxy)-3-(methylseleno)propanoic acid; 2-(3-chloropropanoyloxy)-3-(methylseleno)propanoic acid; 2-{(1H-imidazoyl-4-ylcarbonyl)oxy)}-3-(methylseleno)propanoic acid; 2-(pivaloyloxy)-3-(methylseleno)propanoic acid methyl ester.

9. A method for preparing the selenium compounds of general formula (I), defined in claim 1, wherein it includes the following steps: a) the reaction of a racemic (DL) oxirane-2-carboxylic acid ester or of one of the enantiomers thereof (D or L) which are commercially available, with: either an alkylselenol R.sup.1SeH, which is itself prepared in situ from an alkali metal salt of alkylselenolate of formula R.sup.1Se-M.sub.1 which is itself obtained by reduction of dialkyl diselenide where M.sub.1 represents an alkali metal atom), which is reacted with ammonium chloride; or a dialkylaluminum alkylselenolate derivative of formula Al(R.sup.1).sub.2SeR.sup.1, which is itself generated in situ from the corresponding trialkylaluminum Al(R.sup.1).sub.3 and elemental selenium Se(0), b) if applicable, one or more of the following reactions or series of reactions: hydrolysis of the ester function, then acidification of the reaction medium in order to obtain the corresponding acids of formula (I) where X=OH; then esterification of the acids of formula (I) or of the alkali metal salts thereof with an alcohol or an alkyl halide in order to obtain the corresponding esters of general formula (I) where X=OR.sup.3, with R.sup.3 as defined above; amidification of the acids of formula (I) with an appropriate amine of formula R.sup.4R.sup.5NH or NH.sub.3 where R.sup.5 is as defined above, in order to obtain the compound of general formula (I) where X=NH.sub.2, NR.sup.4R.sup.5 or -amino acid, CH.sub.3SeCH.sub.2CH.sub.2CH(COOH)NH, esterification, when R.sup.2=H, of the hydroxyl function by an appropriate acid in order to obtain the compound of general formula (I) where OR.sup.2 is different from the OH group; salification by an acid or by a base.

10. The method according to claim 9, wherein the selenium reagent is: either a dialkylaluminum alkylselenolate, in an aprotic polar solvent; or an alkylselenol, generated in situ from metal selenium Se(0) and alkyl lithium, in an aprotic polar solvent, and then put in the presence of ammonium chloride.

11. The method according to claim 9, wherein reaction 1) takes place in an aprotic polar solvent, and in that the subsequent reactions leading to the different compounds of formula (I) include at least one acidification, or esterification, or amidification and salification.

12. A method of carrying out treatment of tumors or cancers comprising: providing at least one selenium compound of formula (I) as defined in claim 1 as a pharmaceutical agent alone or combined with at least one other agent selected from at least one other pharmaceutical agent and at least one other antitumor agent, and carrying out a treatment of the tumors or cancers using the pharmaceutical agent alone or combined with at least one other agent selected from at least one other pharmaceutical agent and at least one other antitumor agent.

13. A pharmaceutical composition, wherein it includes at least one pharmaceutically active ingredient including at least one selenium compound of general formula (I) as defined in claim 1, alone or combined with at least one other pharmaceutically active ingredient.

14. The pharmaceutical composition according to claim 13, wherein the selenium compound of general formula (I) constitutes a pharmaceutically active ingredient for carrying out the treatment of tumors or cancers selected from the group consisting of tumors or cancers of the prostate, of the liver, of the pancreas, of the breast, and of the colon, either alone or in combination with one or more other known anticancer or cytotoxic agents, either by pre-administration or by co-administration.

15. The pharmaceutical composition according to claim 13, wherein at least one of the other anticancer agents is selected from the following compounds: aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, matrix metalloproteinase inhibitors; VEGF inhibitors selected from the group comprising anti-VEGF antibodies (Avastin (R)) and the small molecules selected from the group comprising ZD6474 and SU6668, vatalanib, BAY-43-9006, SU11248, CP-547632 and CEP-7055, the SA-1 and SA-2 inhibitors selected from the group comprising the anti-HER2 antibodies (Herceptin), EGFR inhibitors selected from the group comprising gefitinib, erlotinib, ABX-EGF, EMD72000, 11F8, and cetuximab, Eg5 inhibitors selected from the group comprising SB-715992, SB-743921, and MKI-833; PAN inhibitors selected from the group comprising canertinib, EKB-569, CI-1033, AEE-788, and XL-647; kinase inhibitors selected from the group comprising 2C4, and GW-572016, dasatinib, bicalutamide, tamoxifen, MAPK kinase inhibitors, PI3 kinase inhibitors, PDGF inhibitors selected from the group comprising imatinib, anti-angiogenic agents and the antivascular agents, receptor and non-receptor tyrosine kinase inhibitors, inhibitors of integrin signaling; tubulin, anticancer agents selected from the group comprising vinblastine, vincristine, vinorelbine, vinflunine, paclitaxel, docetaxel, 7-O-methylthiomethylpaclitaxel, 4-deacetyl-4-methylcarbonatepaclitaxel, C-4 methyl carbonate paclitaxel, epothilone A, epothilone B, epothilone C, epothilone D, deoxyepothilone A, deoxyepothilone B, oxabicyclo[14.1.0]-heptadecane-5-9-dione (ixabepilone), leucovorin, tegafur, CDK inhibitors, antiproliferative cell cycle inhibitors, epidophyllotoxin, etoposide, VM-26; anti-neoplastic enzymes, topoisomerase I or II inhibitors selected from the group comprising camptothecin, topotecan, SN-38, procarbazine, mitoxantrone; platinum coordination complexes selected from the group comprising cisplatin, carboplatin and oxaliplatin, purine antagonists selected from the group comprising 6-thioguanine and 6-mercaptopurine, and glutamine antagonists.

16. The pharmaceutical composition according to claim 13, wherein at least one of the other cytotoxic agents is selected from at least one compound that follows: cyclophosphamide, doxorubicin, daunorubicin, mitoxantrone, melphalan, hexamethyl melamine, thiotepa, cytarabine, idatrexate, trimetrexate, dacarbazine, L-asparaginase, bicalutamide, leuprolide, pyridobenzoindole derivatives, the interferons, the interleukins.

17. The pharmaceutical composition according to claim 13, wherein it includes at least one selenium compound of formula (I) at a concentration between 0.02% and 0.15% by weight in selenium equivalent.

18. The pharmaceutical composition according to claim 13, wherein it includes at least one pharmaceutically acceptable carrier and a therapeutically effective amount of one or more of the selenium compounds selected from the group consisting of a selenium compound as defined in claim 1, of a stereoisomer thereof, of a tautomer thereof, and of a pharmaceutically acceptable salt thereof, said carrier consisting essentially of: an injectable or drinkable solution, a solid medium composed of one or more excipients which are selected from the group consisting of vitamins, natural antioxidants, L-ergothioneine, mineral salts, mono-, di- or polysaccharides, folic acid, vitamins B.sub.6, vitamin E, vitamin C, lactose, starch.

19. The pharmaceutical composition according to claim 13, wherein it is formulated for a route of administration selected from the oral route, intravenous route, parenteral route, topical route selected from a transdermal route, a nasal route, an ocular route, and inhalation.

20. The pharmaceutical composition according to claim 13, wherein said at least one selenium compound of formula (I) is selected from the group consisting of ##STR00144##

21. The pharmaceutical composition according to claim 20, wherein said at least one selenium compound of formula (I) is incorporated in an effective amount to treat a tumor or cancer selected from the group consisting of a tumor or cancer of the prostate, a tumor or cancer of the liver, a tumor or cancer of the pancreas, a tumor or cancer of the breast, and a tumor or cancer of the colon.

22. The method of claim 12, wherein said at least one selenium compound of formula (I) is selected from the group consisting of ##STR00145##

23. The method according to claim 22, wherein said at least one selenium compound of formula (I) is incorporated in an effective amount to treat a tumor or cancer selected from the group consisting of a tumor or cancer of the prostate, a tumor or cancer of the liver, a tumor or cancer of the pancreas, a tumor or cancer of the breast, and a tumor or cancer of the colon.

Description

DESCRIPTION OF THE FIGURES

(1) FIG. 1: Diagram of the process of synthesis of the compounds according to general formula (I)

(2) FIG. 2: Viability percentage of cells DU145 and LS174T versus the concentration of compound 4, after 96 hours of treatment

(3) FIG. 3: Viability percentage of cells DU145, LS174T and HT-29 versus the concentration of compound 10, after 96 hours of treatment

(4) FIG. 4: Viability percentage of cells PC3, DU145, PANC-1 and MIA PaCa-2 versus the concentration of compound 38, after 96 hours of treatment

(5) FIG. 5: Viability percentage of cells HT-29, LS174T, HepG2 and MCF-7 versus the concentration of compound 38, after 96 hours of treatment

EXAMPLES

(6) The following examples, as well as the diagram of the method according to the invention (FIG. 1) and the other figures, are provided only for the purpose of illustration and can in no way limit the scope of the invention.

(7) In the examples described below, all the percentages are given in weight, the temperature is the ambient temperature or given in degree Celsius, and the pressure is atmospheric pressure unless otherwise indicated.

(8) The used reagents are as commercially available from international suppliers such as SAF (France), Alfa Aesar, Fisher Scientific, TCI Europe, Bachem (Switzerland), except for the following compounds, which were prepared according to the protocol cited: oxirane-2-carboxylate ethyl ester (according to Org. Synth. 2006, 83, 162-169); oxirane-2-carboxylate tert-butyl ester (according to J. Am. Chem. Soc. 2008, 130 (31), 10096-10102), and (R)-oxirane-2-carboxylate benzyl ester (J. Org. Chem. 1992, 57 (11), 3380-3387).

(9) I. Preparation Examples of the Compounds According to the Invention

(10) 1aPreparations of Compounds a by the Introduction of Selenium: 3-(Alkylseleno)-2-Hydroxypropanoic Acid Esters

(11) The compounds A are prepared by reacting an alkylselenol R.sup.1SeH or a dialkylaluminum alkylselenolate Al(R.sup.1).sub.2SeR.sup.1 (generated in situ from trialkyl aluminum and elemental selenium Se(0) according to A. P. Kozikowski and A. Ames, J. Org. Chem. 1978, 43, 2735), with an alkyl oxirane carboxylate.

(12) ##STR00003##

Example A1

Preparation of the methyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 1) using dimethylaluminum methylselenolate

(13) ##STR00004##

(14) 8.6 g (12.6 mL; 25.2 mmol; 1.1 equiv.) of a 2 M solution of trimethylaluminum in toluene are added dropwise (duration of addition 15 min) under nitrogen to 2.0 g (25.2 mmol; 1.1 equiv.) of selenium Se(0). The suspension is stirred for 15 min at ambient temperature, then for 2 h at reflux in a closed environment. The environment is allowed to return to ambient temperature and then cooled to 0 C. under nitrogen.

(15) 2.386 g (22.9 mmol) of methyl oxirane-2-carboxylate in solution in 12 mL of dichloromethane are added dropwise to the reaction medium (duration of addition 15 min). The medium is left under stirring for 30 min at 0 C. and then for 16 h at ambient temperature.

(16) The reaction medium is cooled to 4 C. for 15 min. 1.286 g (24.05 mmol; 1.05 equiv.) of ammonium chloride in 10 mL of water are added very slowly dropwise to the reaction medium (duration of addition 15 min), because a very strong gaseous evolution occurs (Attention: generation and evolution of methane). 20 mL of dichioromethane are added dropwise, then the medium is stirred for 10 min without heating. 20 mL of a saturated NH.sub.4Cl aqueous solution are added dropwise, then the medium is stirred for 10 min without heating.

(17) The reaction medium is filtered on Celite which is rinsed with dichloromethane (720 mL). The organic phase is decanted, and the aqueous phase is extracted with dichloromethane (320 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated.

(18) The yellow oil obtained is distilled at a reduced pressure of 8 mbar (125 C.). The yield consists of 1.998 g (42%) of compound 1 in the form of a yellow oil.

(19) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.09 (s, 3H); 2.91 (m, 1H); 3.01 (m, 1H); 3.18 (m, 1H); 3.83 (s, 3H); 4.51 (m, 1H).

(20) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=5.9; 29.9; 53.1; 70.7; 174.1

(21) UPLC-MS (AP+): 220.8 (M+Na).sup.+

Example A2

Preparation of the methyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 1) using methylselenol

(22) ##STR00005##

(23) 992 mg (12.5 mmol; 1 eq.) of selenium Se(0) are suspended under nitrogen in 44 mL of THF. The suspension is cooled to 3 C., then 6.2 mL (18.6 mmol; 1.49 eq.) of a 3 M methyllithium solution in diethoxymethane are added dropwise (addition time 7 min). The completely discolored medium is stirred for 20 min without heating, then 802 mg (15 mmol; 1.2 eq.) of ammonium chloride dissolved in 44 mL of methanol are added dropwise. The medium is stirred for 20 min without heating, then 2.11 mL (15 mmol; 1.2 eq.) of triethylamine are added. The medium is stirred for 20 min without heating, then 1.71 g (16.25 mmol; 1.3 eq.) of methyl oxirane-2-carboxylate are added. The medium is stirred for 1 h at 0 C., then for 22 h at ambient temperature.

(24) The reaction medium is cooled to 0 C. for 15 min. 90 mL of dichloromethane are added dropwise, then the medium is stirred for 10 min without heating. 90 mL of a saturated NH.sub.4Cl aqueous solution are added dropwise, then the medium is stirred for 10 min without heating. The medium is diluted with 90 mL of water. The organic phase is recovered, and the aqueous phase is extracted with dichloromethane (290 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated. The yellow oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(25) The yield consists of 330 mg (14%) of compound 1.

Example A3

Preparation of the methyl ester of (R)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 2)

(26) ##STR00006##

(27) Compound 2 is obtained using the conditions of Example A1, starting with 4.959 g of methyl (S)-oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(28) The yield consists of 3.257 g (33%) of compound 2 in the form of a yellow oil.

(29) The .sup.1H NMR spectrum is identical to the one obtained in Example A1.

(30) [].sub.D: 11.5 (c=1.0; MeOH)

Example A4

Preparation of the methyl ester of (S)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 3)

(31) ##STR00007##

(32) Compound 3 is obtained using the conditions of Example A1, starting with 7 g of methyl (R)-oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(33) The yield consists of 4.04 g (29%) of compound 3 in the form of an orange oil.

(34) The .sup.1H NMR spectrum is identical to the one obtained in Example A1.

(35) [].sub.D: 11.1 (c=1.0; MeOH)

Example A5

Preparation of the ethyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 4)

(36) ##STR00008##

(37) Compound 4 is obtained using the conditions of Example A1, starting with 1.56 g of ethyl oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(38) The yield consists of 1.188 g (38%) of compound 4 in the form of a yellow oil.

(39) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.35 (t, J=7.0, Hz, 3H); 2.10 (s, 3H); 2.91 (dd, J=13.0, Hz, J=5.5 Hz, 1H); 3.01 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 3.16 (d, J=6.0 Hz, 1H); 4.3 (q, J=7.0 Hz, 2H); 4.5 (td, J=5.5 Hz, J=4.0 Hz, 1H).

(40) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=6.0; 14.6; 29.9; 62.4; 70.8; 173.7

(41) UPLC-MS (AP+): 234.8 (M+Na).sup.+

Example A6

Preparation of the tert-butyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 5)

(42) ##STR00009##

(43) Compound 5 is obtained using the conditions of Example A1, starting with 1.0 g of tert-butyl oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(44) The yield consists of 889 mg (54%) of compound 5 in the form of a slightly yellow oil.

(45) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.53 (s, 9H); 2.13 (s, 3H); 2.87 (dd, J=13.0 Hz, J=5.5 Hz, 1H); 2.97 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 3.19 (d, J=5.5 Hz, 1H); 4.39 (td, J=5.5 Hz, J=4.0 Hz, 1H).

Example A7

Preparation of the benzyl ester of (S)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 6)

(46) ##STR00010##

(47) Compound 6 is obtained using the conditions of Example A1, starting with 622 mg of benzyl (R)-oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(48) The yield consists of 325 mg (33%) of compound 6 in the form of a slightly yellowish oil.

(49) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.05 (s, 3H); 2.91 (dd, J=13.0 Hz, J=5.5 Hz, 1H); 3.02 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.18 (d, J=5.5 Hz, 1H); 4.55 (td, J=5.5 Hz, J=4.5 Hz, 1H), 5.26 (s, 2H); 7.41 (m, 5H).

Example A8

Preparation of the methyl ester of 3-(ethylseleno)-2-hydroxypropanoic acid (Compound 7)

(50) ##STR00011##

(51) Compound 7 is obtained using the conditions of Example A1, starting with 2 g of selenium Se(0), 10.98 g of a 25% triethylaluminum solution in toluene and 2.386 g of methyl oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(52) The yield consists of 1.977 g (40%) of compound 7 in the form of a yellow oil.

(53) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.41 (t, J=7.5 Hz, 3H); 2.67 (q, J=7.5 Hz, 2H); 2.92 (dd, J=13.0 Hz, J=5.5 Hz, 1H); 3.02 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.18 (d, J=4.5 Hz, 1H); 3.82 (s, 3H), 4.5 (m, 1H).

(54) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm): 16.2; 18.9; 27.8; 53.1; 70.8; 174.1

Example A9

Preparation of the methyl ester of 2-hydroxy-3-(isobutylseleno)propanoic acid (Compound 8)

(55) ##STR00012##

(56) Compound 8 is obtained using the conditions of Example A1, starting with 2.0 g of selenium Se(0), 19.08 g of a 25% solution of triisobutyl aluminum in toluene and 2.386 g of methyl oxirane-2-carboxylate. After rinsing the Celite and evaporation of the solvents, the oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(57) The yield consists of 2.291 g (42%) of compound 8 in the form of a yellow oil.

(58) .sup.1H NMR (CDCl.sub.3, 300 MHz): (ppm)=0.77 (d, J=6.5 Hz, 6H); 1.61 (m, 11-1); 2.39 (d, J=7.0 Hz, 2H); 2.66 (dd, J=13.0 Hz, J=5.5 Hz, 1H); 2.76 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.58 (s, 3H); 4.25 (m, 1H).

(59) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=22.9; 29.0; 29.7; 35.7; 53.1; 70.7; 174.0

(60) 1bPreparation of the Compounds a by Esterification

(61) The Compounds A can be obtained by introducing selenium reagents, as described in paragraph 1a above, but also by esterification of the Compounds B (for their preparation, see paragraph 2 below).

(62) ##STR00013##

Example A10

Preparation of the ethyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 4)

(63) ##STR00014##

(64) 500 mg (2.68 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10, paragraph 2) are dissolved in 11 mL of absolute ethanol under nitrogen. 66 mg (1.07 mmol; 0.4 eq.) of orthoboric acid are added to the medium. The medium is left at reflux under stirring and under nitrogen for 48 h.

(65) 33 mg (535 mol; 0.2 eq.) of orthoboric acid are added again to the medium.

(66) The medium is left at reflux for 24 h.

(67) The medium is concentrated to dryness, then the concentrate is redissolved with a semi-saturated NH4Cl aqueous solution (40 mL). The medium is extracted with ethyl acetate (340 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated.

(68) The yield consists of 446 mg (77%) of compound 4 in the form of a yellow oil.

(69) The .sup.1H NMR spectrum is identical to the one obtained in Example A5.

Example A11

Preparation of the isopropyl ester of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 9)

(70) ##STR00015##

(71) 374 mg (2 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved in 8 mL of 2-propanol under nitrogen. 49 mg (800 mol; 0.4 eq.) of orthoboric acid are added to the medium. The medium is left at reflux under stirring and under nitrogen for 48 h.

(72) 25 mg (400 mol; 0.2 eq.) of orthoboric acid are added again to the medium.

(73) The medium is left at reflux for 24 h.

(74) The medium is concentrated to dryness, then the concentrate is redissolved with a semi-saturated NaHCO.sub.3 aqueous solution (40 mL). The medium is extracted with ethyl acetate (340 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated. The oil obtained is purified on a silica column (cyclohexane/ethyl acetate).

(75) The yield consists of 285 mg (62%) of compound 9 in the form of a yellow oil.

(76) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.32 (d, J=6.0 Hz, 6H); 2.11 (s, 3H); 2.89 (dd, J=13.0 Hz, J=5.5 Hz, 1H); 2.99 (m, 1H); 3.20 (s, 1H); 4.45 (m, 1H); 5.14 (m, 1H).

(77) .sup.13C NMR (CDCl.sub.3 75 MHz): (ppm)=6.0; 22.2; 29.8; 70.4; 70.9; 173.2

(78) UPLC-MS (AP+): 248.9 (M+Na).sup.+

(79) 2Preparation of the Compounds B and C: the 3-(alkylseleno)-2-hydroxypropanoic Acids and the Corresponding Salts

(80) The Compounds B are prepared by hydrolysis of the ester function of the Compounds A, and the corresponding salts C are obtained by reacting the compounds B with oxides or hydroxides:

(81) ##STR00016##

Example B1

Preparation of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 10) from the methyl ester

(82) ##STR00017##

(83) 4.908 g (23.53 mmol) of compound 1 are dissolved in 14 mL of THF, 5 mL of methanol and 5 mL of demineralized water. 47.1 mL (47.07 mmol; 2 equiv.) of 1 M lithium hydroxide aqueous solution are added, the solution is stirred at ambient temperature for 16 h.

(84) The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid solution (14 mL). The medium is extracted with ethyl acetate (4100 mL). The organic phases are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(85) The yield consists of 4.177 g (95%) of compound 10 in the form of a pale yellow solid.

(86) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.08 (s, 3H); 2.92 (dd, J=13.5 Hz, J=6.5 Hz, 1H); 3.03 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.57 (dd, J=6.5 Hz, J=4.5 Hz, 1H).

(87) .sup.1H NMR (DMSO, 400 MHz): (ppm)=1.99 (s, 3H); 2.70 (dd, J=12.5 Hz, J=6.5 Hz, 1H); 2.79 (dd, J=12.5 Hz, J=5.0 Hz, 1H); 4.18 (dd, J=6.5 Hz, J=5.0 Hz, 1H).

(88) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=6.0; 29.6; 70.1; 178.0

(89) UPLC-MS (AP): 182.6 (MH.sup.+)

(90) Elemental analysis: C.sub.4H.sub.8O.sub.3Se; Theoretical: C (26.24%); H (4.4%); Experimental: C (26.9%); H (4.42%)

Example B2

Preparation of 2-hydroxy-3-(methylseleno)propanoic acid (Compound 10) from the tert-butyl ester

(91) The compound 10 is obtained using the conditions of Example B1, starting with 100 mg of tert-butyl 2-hydroxy-3-(methylseleno)propanoate (5).

(92) The yield consists of 64 mg (82%) of the desired product in the form of a solid.

(93) The .sup.1H NMR spectrum is identical to the one obtained in Example B1.

Example B3

Preparation of (R)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 11)

(94) ##STR00018##

(95) Compound 11 is obtained using the conditions of Example B1, starting with 2.117 g of methyl (R)-2-hydroxy-3-(methylseleno)propanoate (2).

(96) The yield consists of 1.875 g (96%) of the desired product in the form of a yellow solid.

(97) The .sup.1H NMR spectrum is identical to the one obtained in Example B1.

(98) [].sub.D=+1.07 (c=6.0; EtOH)

Example B4

Preparation of (S)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 12) from the methyl ester

(99) ##STR00019##

(100) Compound 12 is obtained using the conditions of Example B1, starting with 2.95 g of methyl (S)-2-hydroxy-3-(methylseleno)propanoate (3).

(101) The yield consists of 2.61 g (95%) of the desired product in the form of a yellow solid.

(102) The .sup.1H NMR spectrum is identical to the one obtained in Example B1.

(103) [].sub.D=1.03 (c=6.0; EtOH)

Example B5

Preparation of (S)-2-hydroxy-3-(methylseleno)propanoic acid (Compound 12) from benzyl ester

(104) ##STR00020##

(105) Compound 12 is obtained using the conditions of Example B1, starting with 494 mg of benzyl (S)-2-hydroxy-3-(methylseleno)propanoate (6).

(106) After 16 h of stirring at ambient temperature, the reaction medium is extracted with ethyl acetate (230 mL). The pH of the medium is adjusted to 1 by adding 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (230 mL). The organic phases of this second extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(107) The yield consists of 280 mg (84%) of compound 12 in the form of a pale yellow solid.

(108) The .sup.1H NMR spectrum is identical to the one obtained in Example B1.

Example B6

Preparation of 3-ethylseleno-2-hydroxypropanoic acid (Compound 13) from the methyl ester

(109) ##STR00021##

(110) Compound 13 is obtained using the conditions of Example B1, starting with 500 mg of 3-(ethylseleno)-2-hydroxypropanoic acid methyl ester (7). The yield consists of 471 mg (100%) of compound 13 in the form of a pale yellow solid.

(111) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.44 (t, J=7.5 Hz, 3H); 2.71 (q, J=7.5 Hz, 2H); 3.00 (dd, J=13.5 Hz, J=6.0 Hz, 1H); 3.11 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.55 (dd, J=6.0 Hz, J=4.5 Hz, 1H).

(112) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=16.1; 19.1; 27.6; 70.3; 177.9

(113) UPLC-MS (AP): 197.2 (MH.sup.+)

Example B7

Preparation of 2-hydroxy-3-(isobutylseleno)propanoic acid (Compound 14) from the methyl ester

(114) ##STR00022##

(115) Compound 14 is obtained using the conditions of Example B1 starting with 500 mg of 2-hydroxy-3-(isobutylseleno)propanoic acid methyl ester (8).

(116) The yield consists of 423 mg (90%) of compound 14 in the form of a pale yellow solid.

(117) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.03 (d, J=6.5 Hz, 6H); 1.88 (m, 1H); 1.95 (dd, J=7.0 Hz, 2H); 2.97 (dd, J=13.0 Hz, J=6.0 Hz, 1H); 3.08 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 4.53 (m, 1H).

(118) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=22.9; 28.7; 29.7; 35.8; 70.2; 177.9

(119) UPLC-MS (AP): 225.3 (MH.sup.+)

Example C1

Preparation of the dicyclohexylammonium 2-hydroxy-3-(methylseleno)propanoate salt (Compound 15)

(120) ##STR00023##

(121) 373 mg (2 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid are dissolved in 2 mL of acetone. 733 mg (805 L; 4 mmol; 2 eq.) of dicyclohexylamine are added to the medium.

(122) A precipitate appeared instantaneously and is filtered, rinsed with acetone (210 mL), then with a 1/1 ethyl acetate/cyclohexane mixture (210 mL).

(123) The yield consists of 524 mg (70%) of compound 15 in the form of a white solid.

(124) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.26 (m, 7H); 1.50 (m, 4H); 1.68 (m, 2H); 1.84 (m, 4H); 2.06 (m, 4H); 2.12 (s, 3H); 2.91 (dd, J=12.5 Hz, J=6. Hz, 1H); 3.02 (m, 3H); 4.22 (m, 1H).

(125) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=5.7; 25.2; 25.5; 29.4; 31.6; 53.2; 72.2; 177.7

Example C2

Preparation of sodium 2-hydroxy-3-(methylseleno)propanoate salt (Compound 16)

(126) ##STR00024##

(127) 80 mg (2 mmol) of 60% sodium hydroxide in mineral oil are suspended under nitrogen in 2 mL of THF. 373 mg (2 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid dissolved in 2 mL of THF are added dropwise in 5 min.

(128) The medium is cooled in a water-ice bath, then cyclohexane (3 mL) is added.

(129) A precipitate forms. The medium is filtered, then the solid is washed with cyclohexane (3 mL), then with TBME (33 mL).

(130) The yield consists of 315 mg (75%) of compound 16 in the form of an off-white solid.

(131) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.08 (s, 3H); 2.88 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 2.99 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.32 (dd, J=6.5 Hz, J=4.0 Hz, 1H).

(132) .sup.13C NMR (D.sub.2O, 75 MHz): (ppm)=4.8; 30.0; 71.7; 179.4

(133) UPLC-MS (AP): 182.7 (MNa.sup.+)

Example C3

Preparation of the magnesium bis(2-hydroxy-3-(methylseleno)propanoate salt (Compound 17)

(134) ##STR00025##

(135) 495 mg (2.65 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid are dissolved in 1 mL of demineralized water, then 50 mg (1.25 mmol) of magnesium oxide are added. The medium is stirred at ambient temperature for 16 h.

(136) The medium is diluted with acetone (2 mL), then the medium is filtered through fritted glass. The solid is rinsed with water (0.5 mL), then with TBME (2 mL). The solid is dried under a vacuum.

(137) The yield consists of 565 mg of compound 17 in the form of a white solid (quantitative).

(138) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.04 (s, 3H); 2.83 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 2.95 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.25 (dd, J=6.5 Hz, J=4.0 Hz, 1H).

Example C4

Preparation of the zinc bis(2-hydroxy-3-(methylseleno)propanoate salt (Compound 18)

(139) ##STR00026##

(140) 500 mg (2.65 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid are dissolved in 1 mL of demineralized water, then 145 mg of basic zinc carbonate are added. 1 mL of demineralized water is added, then the medium is stirred at ambient temperature for 16 h.

(141) The medium is diluted with acetone (2 mL), then the medium is filtered through fritted glass. The solid is rinsed with water (1 mL), then with acetone (32 mL). The solid is dried under a vacuum.

(142) The yield consists of 496 mg (79%) of compound 18 in the form of a white solid.

(143) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.05 (s, 3H); 2.87 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 2.98 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.32 (dd, J=6.5 Hz, J=4.0 Hz, 1H).

Example C5

Preparation of the calcium bis(2-hydroxy-3-(methylseleno)propanoate salt (Compound 19)

(144) ##STR00027##

(145) 374 mg (2 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid are dissolved in 0.9 mL of demineralized water, then 70 mg (945 mol) of calcium hydroxide are added. 1.1 mL of demineralized water are added, then the medium is stirred at ambient temperature for 16 h.

(146) The medium is filtered through fritted glass. The solid is rinsed with water (22 mL), then with tert-butyl methyl ether (TBME) (22 mL). The solid is dried under a vacuum. The yield consists of 74 mg (19%) of the desired product in the form of an off-white solid.

(147) The filtrate is concentrated to dryness. The residue is triturated with TBME (5 mL). The solid is dried under a vacuum.

(148) The yield consists of 205 mg (53%) of compound 19 in the form of an off-white solid.

(149) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.07 (s, 3H); 2.88 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 2.99 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.30 (dd, J=6.5 Hz, J=4.0 Hz, 1H).

(150) .sup.13C NMR (D.sub.2O, 300 MHz): (ppm)=4.8; 30.4; 72.1; 180.1

(151) 3Preparation of Compounds D: Amides Derived from 2-hydroxy-3-(alkylseleno)propanoic acid

(152) The Compounds D are prepared either by aminolysis of compounds A, or they are synthesized from compounds B by peptide coupling.

(153) ##STR00028##

Example D1

Preparation of 2-hydroxy-3-(methylseleno)propanamide (Compound 20)

(154) ##STR00029##

(155) 295 mg (1.5 mmol) of compound 1 are dissolved in 4.04 g (5.25 mL; 36.76 mmol; 24 equiv.) of a 20% aqueous ammonia solution. The medium is stirred at ambient temperature for 22 h.

(156) The medium is evaporated to dryness then coevaporated with ethyl acetate.

(157) The yield consists of 284 mg (100%) of compound 20 in the form of an off-white solid.

(158) .sup.1H NMR (MeOD, 400 MHz): (ppm): 2.11 (s, 3H); 2.85 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 2.98 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.3 (dd, J=7.0 Hz, J=4.0 Hz, 1H).

(159) .sup.1H NMR (DMSO, 400 MHz): (ppm)=1.97 (s, 3H); 2.68 (dd, J=12.5 Hz, J=7.0 Hz, 1H); 2.80 (dd, J=12.5 Hz, J=4.0 Hz, 1H); 4.04 (m, 1H); 5.58 (d, J=5.5 Hz, 1H); 7.19 (d, J=15.5 Hz, 2H)

(160) .sup.13C NMR (MeOD, 75 MHz): (ppm): 5.2; 31.0; 73.3; 179.2

(161) UPLC-MS (AP+): 183.8 (M+H).sup.+

(162) UPLC-MS (AP+): 206.0 (M+Na).sup.+

Example D2

Preparation of N-cyclopropyl-2-hydroxy-3-(methylseleno)propanamide (Compound 21)

(163) ##STR00030##

(164) 513 mg (2.5 mmol) of compound 1 are dissolved in 428 mg (519 L; 7.5 mmol; 3 equiv.) of cyclopropylamine. The medium is stirred at 85 C. for 72 h.

(165) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (dichioromethane/methanol).

(166) The yield consists of 452 mg (79%) of compound 21 in the form of a pale yellow solid.

(167) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=0.56 (m, 2H); 0.82 (m, 2H); 2.03 (s, 3H); 2.76 (m, 1H); 2.89 (dd, J=13.0 Hz, J=7.5 Hz, 1H); 3.08 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.52 (d, J=3.5 Hz, 1H); 4.17 (m, 1H); 6.88 (s, 1H).

(168) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=4.9; 6.8; 6.9; 22.7; 31.6; 69.9; 173.8

(169) UPLC-MS (AP+): 245.9 (M+Na).sup.+

Example D3

Preparation of N-[2-(dimethylamino)ethyl]-2-hydroxy-3-(methylseleno)propanamide (Compound 22)

(170) ##STR00031##

(171) 417 mg (2 mmol) of compound 1 are dissolved in 539 mg (674 L; 6 mmol; 3 equiv.) of N,N-dimethylethylenediamine. The medium is stirred at 85 C. for 72 h.

(172) The medium is allowed to return to ambient temperature, evaporated to dryness then purified on a silica column (dichloromethane/methanol).

(173) The yield consists of 460 mg (89%) of compound 22 in the form of an orange oil.

(174) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.05 (s, 3H); 2.28 (s, 6H); 2.48 (t, J=6.0 Hz, 2H); 2.91 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 3.06 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 3.40 (m, 2H); 4.26 (dd, J=7.0 Hz, J=4.0 Hz, 1H); 7.34 (m, 1H).

(175) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=5.2; 31.6; 36.7; 45.6; 58.6; 71.0; 172.9

(176) UPLC-MS (AP+): 254.9 (M+H).sup.+

Example D4

Preparation of 2-hydroxy-3-(methylseleno)-1-(pyrrolidin-1-yl)propan-1-one (Compound 23)

(177) ##STR00032##

(178) 513 mg (2.5 mmol) of compound 1 are dissolved in 533 mg (616 L; 7.5 mmol; 3 equiv.) of pyrrolidine. The medium is stirred at 85 C. for 48 h.

(179) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (cyclohexane/ethyl acetate).

(180) The yield consists of 402 mg (66%) of compound 23 in the form of an orange oil.

(181) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.85-2.07 (m, 4H); 2.11 (s, 3H); 2.77 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 2.85 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.45-3.65 (m, 4H); 3.78 (d, J=8.0 Hz, 1H); 4.45 (m, 1H).

(182) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=5.8; 24.3; 26.5; 29.7; 46.7; 46.8; 70.2; 171.6

(183) UPLC-MS (AP+): 237.9 (M+H).sup.+

(184) UPLC-MS (AP+): 259.9 (M+Na).sup.+

Example D5

Preparation of 2-hydroxy-3-(methylseleno)-1-(piperidin-1-yl)propan-1-one (Compound 24)

(185) ##STR00033##

(186) 417 mg (2 mmol) of compound 1 are dissolved in 517 mg (600 L; 6 mmol; 3 equiv.) of piperidine. The medium is stirred at 85 C. for 72 h.

(187) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (dichloromethane/acetone).

(188) The yield consists of 151 mg (29%) of compound 24 in the form of a yellow oil.

(189) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.56-1.74 (m, 6H); 2.13 (s, 3H); 2.72 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 2.82 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 3.40 (d, J=4.0 Hz, 2H); 3.62 (m, 2H); 4.03 (d, J=7.0 Hz, 1H); 4.61 (m, 1H).

(190) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=6.0; 24.8; 25.8; 26.6; 30.4; 44.2; 46.5; 68.7; 171.3

(191) UPLC-MS (AP+): 251.9 (M+H).sup.+

(192) UPLC-MS (AP+): 273.9 (M+Na).sup.+

Example D6

Preparation of 2-hydroxy-3-(methylseleno)-1-(morpholin-4-yl)propan-1-one (Compound 25)

(193) ##STR00034##

(194) 486 mg (2.4 mmol) of compound 1 are dissolved in 641 L (7.2 mmol; 3 equiv.) of morpholine. The medium is stirred at 85 C. for 72 h.

(195) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (dichloromethane/acetone).

(196) The yield consists of 262 mg (42%) of compound 25 in the form of a pale yellow oil.

(197) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.12 (s, 3H); 2.79 (m, 2H); 3.52 (m, 2H); 3.65 (m, 1H); 3.74 (m, 5H); 3.82 (d, J=8.0 Hz, 1H); 4.59 (m, 1H).

(198) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=6.1; 30.1; 43.3; 46.2; 66.8; 67.1; 68.6; 171.8

(199) UPLC-MS (AP+): 253.9 (M+H).sup.+

(200) UPLC-MS (AP+): 276.0 (M+Na).sup.+

Example D7

Preparation of N,N-diethyl-2-hydroxy-3-(methyl seleno)propanamide (Compound 26)

(201) ##STR00035##

(202) 417 mg (2 mmol) of compound 1 are dissolved in 443 mg (633 L; 6 mmol; 3 equiv.) of diethylamine. The medium is stirred at 85 C. for 72 h.

(203) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (dichloromethane/methanol).

(204) The yield consists of 105 mg (19%) of compound 26 in the form of an orange oil.

(205) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.38 (t, J=7.0 Hz, 6H); 2.10 (s, 3H); 2.91 (dd, J=12.5 Hz, J=6.0 Hz, 1H); 3.02 (m, 5H); 4.33 (m, 1H).

Example D8

Preparation of 2-hydroxy-N-(2-hydroxyethyl)-3-(methylseleno)propanamide (Compound 27)

(206) ##STR00036##

(207) 250 mg (1.22 mmol) of compound 1 are dissolved in 228 mg (225 L; 3.66 mmol, 3 equiv.) of ethanolamine. The medium is stirred at 85 C. for 72 h.

(208) The medium is allowed to return to ambient temperature, evaporated to dryness, then purified on a silica column (dichloromethane/methanol).

(209) The yield consists of 203 mg (72%) of compound 27 in the form of a greenish oil.

(210) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.06 (s, 3H); 2.92 (dd, J=13.0 Hz, J=7.5 Hz, 1H); 2.95 (s, 1H); 3.11 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 3.49 (m, 2H); 3.65 (d, J=4.5 Hz, 1H); 3.78 (m, 2H); 4.26 (m, 1H); 7.26 (s, 1H).

(211) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm)=5.2; 31.2; 42.5; 62.2; 70.6; 173.9

(212) UPLC-MS (AP+): 227.9 (M+H).sup.+

(213) UPLC-MS (AP+): 249.8 (M+Na).sup.+

Example D9

Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-alanine (Compound 29)

D9.1. Preparation of the tert-butyl ester of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-alanine (Compound 28)

(214) ##STR00037##

(215) 100 mg (546 mol) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved under nitrogen in 5 mL of dichloromethane. The medium is cooled to 0 C., then 183 mg (600 mol, 1.1 eq.) of 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one are added. The medium is stirred for 30 min at 0 C., then for 1 h 30 at ambient temperature. The medium is cooled to 0 C., then 99 mg (546 mol, 1 eq.) of the hydrochloride of the tert-butyl ester of (S)-alanine and 156 mg (200 L; 1.20 mmol; 2.2 eq.) of N,N-diisopropylethylamine are added. The medium is stirred for 30 min at 0 C., then for 16 h at ambient temperature.

(216) The reaction medium is diluted with dichloromethane (25 mL), then the medium is washed with a 1N hydrochloric acid aqueous solution (210 mL), then with an aqueous solution of NaHCO.sub.3 (1N, 210 mL), then with a saturated NaCl aqueous solution (10 mL). The organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(217) The yield consists of 116 mg (67%) of compound 28 in the form of a colorless oil.

(218) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 28a (R,S) and 28b (S,S): (ppm)=1.42 and 1.43 (2d, J=3.5 Hz, 3H); 1.49 and 1.50 (2s, 9H); 2.04 and 2.05 (2s, 3H); 2.92 (dd, J=13.0 Hz, J=7.5 Hz, 1H); 3.09 (m, 1H); 3.43 (d, J=3.5 Hz, 1H); 4.21 (m, 1H); 4.48 (m, 1H); 7.27 and 7.33 (2d, J=6.5 Hz, 1H).

(219) UPLC-MS (AP): 309.8 (MH+)

(220) UPLC-MS (AP+): 333.9 (M+Na).sup.+

D9.2: Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-alanine (Compound 29)

(221) ##STR00038##

(222) Compound 29 is obtained using the conditions of Example B1, starting with 58 mg of the ester 28.

(223) The yield consists of 40 mg (81%) of compound 29 in the form of a colorless oil.

(224) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 29a (R,S) and 29b (S,S): (ppm)=1.51 and 1.53 (2d, J=3.0 Hz, 3H); 2.05 and 2.06 (2s, 3H); 2.90 (m, 1H); 3.10 (m, 1H); 3.52 (s, 1H); 4.24 and 4.31 (2dd, J=8.0 Hz, J=4.0 Hz, 1H); 4.61 (m, 1H); 7.36 and 7.43 (2d, J=7.5 Hz, 1H).

Example D10

Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31)

D10.1. Preparation of the methyl ester of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 30)

(225) ##STR00039##

(226) Compound 30 is obtained using the conditions of Example D9, starting with 500 mg (2.65 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) and 557 mg (2.65 mmol; 1 eq.) of the methyl ester of the hydrochloride of (S)-methionine. The yield consists of 549 mg (60%) of the desired product in the form of a slightly yellow oil.

(227) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 30a (R,S) and 30b (S,S): (ppm)=2.05 (m, 4H); 2.13 (m, 3H); 2.23 (m, 1H); 2.56 (t, J=7.5 Hz, 2H); 2.92 (m, 1H); 3.09 (m, 1H); 3.45 (m, 1H); 3.79 and 3.80 (2s, 3H); 4.25 (m, 1H); 4.75 (m, 1H); 7.39 (m, 1H).

D10.2: Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31)

(228) ##STR00040##

(229) Compound 31 is obtained using the conditions of Example B1, starting with 445 mg of compound 30.

(230) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (40 mL), then extracted with ethyl acetate (225 mL). The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (425 ml). The organic phases of this second extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(231) The yield consists of 400 mg (92%) of the desired product in the form of a pale yellow oil.

(232) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 31a (R,S) and 31b (S,S): (ppm)=2.05 and 2.08 (2s, 3H); 2.11 (m, 1H); 2.15 (s, 3H); 2.27 (m, 1H); 2.62 (t, J=7.5 Hz, 2H); 2.91 (m, 1H); 3.10 (m, 1H); 4.28 and 4.38 (2dd, J=8.0 Hz, J=4.5 Hz, 1H); 4.75 (m, 1H); 7.51 and 7.57 (2d, J=8.0 Hz, 1H).

Example D11

Preparation of [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31a)

D11.1 Preparation of the methyl ester of [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 30a)

(233) ##STR00041##

(234) Compound 30a is obtained using the conditions of Example D9, starting with 100 mg (513 mol; 1 eq.) of (R)-2-hydroxy-3-(methylseleno)propanoic acid (11) and 108 mg (513 mol; 1 eq.) of the methyl ester of the hydrochloride of (S)-methionine. The yield consists of 89 mg (50%) of the desired product in the form of a colorless oil.

(235) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.06 (s, 3H); 2.07 (m, 1H); 2.13 (s, 3H); 2.22 (m, 1H); 2.57 (t, J=7.5 Hz, 2H); 2.93 (dd, J=13.0 Hz, J=7.5 Hz, 1H); 3.09 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.40 (d, J=4.0 Hz, 1H); 3.80 (s, 3H); 4.28 (m, 1H); 4.75 (m, 1H); 7.40 (d, J=8.0 Hz, 1H).

D11.2. Preparation of [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31a)

(236) ##STR00042##

(237) Compound 31a is obtained using the conditions of Example D10.2, starting with 89 mg of compound 30a.

(238) The yield consists of 82 mg (99%) of the desired product in the form of a colorless oil.

(239) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.08 (s, 3H); 2.11 (m, 1H); 2.15 (s, 3H); 2.28 (m, 1H); 2.63 (m, 2H); 2.91 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 3.08 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.39 (dd, J=7.0 Hz, J=4.0 Hz, 1H); 4.74 (m, 1H); 7.59 (d, J=8.0 Hz, 1H).

Example D12

Preparation of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31b)

D12.1. Preparation of the methyl ester of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 30b)

(240) ##STR00043##

(241) Compound 30b is obtained using the conditions of Example D9, starting with 100 mg (535 mol; 1 eq.) of (S)-2-hydroxy-3-(methylseleno)propanoic acid (12) and 112 mg (535 mol; 1 eq.) of the methyl ester of the hydrochloride of (S)-methionine. The yield consists of 115 mg (57%) of the desired product in the form of a colorless oil.

(242) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.04 (s, 3H); 2.07 (m, 1H); 2.13 (s, 3H); 2.23 (m, 1H); 2.57 (t, J=7.5 Hz, 2H); 2.92 (dd, J=13.0 Hz, J=8.0 Hz, 1H); 3.10 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.42 (d, J=4.5 Hz, 1H); 3.79 (s, 3H); 4.22 (m, 1H); 4.75 (m, 1H); 7.35 (d, J=8.0 Hz, 1H).

D12.2. Preparation of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-methionine (Compound 31b)

(243) ##STR00044##

(244) Compound 31 b is obtained using the conditions of Example D10.2, starting with 115 mg of compound 30b.

(245) The yield consists of 98 mg (98%) of the desired product in the form of a yellow oil.

(246) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.05 (s, 3H); 2.10 (m, 1H); 2.15 (s, 3H); 2.28 (m, 111); 2.62 (t, J=7.5 Hz, 2H); 2.91 (dd, J=13.0 Hz, J=8.0 Hz, 1H); 3.11 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.27 (dd, J=8.0 Hz, J=4.5 Hz, 1H); 4.75 (m, 1H); 7.48 (d, J=8.0 Hz, 1H).

Example D13

Preparation of the methyl ester of N()-[(2RS)-2-hydroxy-3-methylselenopropanoyl]-N()-tert-butoxycarbonyl-(S)-lysine (Compound 32)

(247) ##STR00045##

(248) Compound 32 is obtained using the conditions of Example D9, starting with 100 mg (535 mol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) and 162 mg (546 mol; 1 eq.) of the methyl ester of the hydrochloride of N()-tert-butoxycarbonyl-(S)-lysine.

(249) The yield consists of 171 mg (73%) of the desired product in the form of a colorless oil.

(250) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers (R,S) and (S,S): (ppm)=1.37 (m, 2H); 1.44 (s, 9H); 1.48 (m, 2H); 1.73 (m, 1H); 1.89 (m, 1H); 2.02 and 2.04 (2s, 3H); 2.89 (m, 1H); 3.07 (m, 3H); 3.74 and 3.75 (s, 3H); 3.85 (m, 1H); 4.26 (m, 1H); 4.59 (m, 1H); 4.69 (m, 1H); 7.30 (m, 1H).

(251) UPLC-MS (AP+): 449.0 (M+Na).sup.+

Example D14

Preparation of the methyl ester of N()-[(2RS)-2-hydroxy-3-methylselenopropanoyl]-N()-fluoroenylmethyloxycarbonyl-(S)-lysine (Compound 33)

(252) ##STR00046##

(253) Compound 33 is obtained using the conditions of Example D9, starting with 200 mg (1.059 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) and 448 mg (1.059 mmol; 1 eq.) of the methyl ester of the hydrochloride of N()-fluorenylmethyloxycarbonyl-(S)-lysine.

(254) The yield consists of 430 mg (59%) of the desired product in the form of a colorless oil.

(255) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.42 (m, 2H); 1.58 (m, 2H); 1.76 (m, 2H); 1.91 (m, 1H); 2.03 (s, 3H); 2.92 (m, 1H); 3.10 (m, 1H); 3.21 (m, 2H); 3.78 (s, 3H); 4.25 (m, 2H); 4.43 (m, 2H); 4.64 (m, 1H); 4.90 (m, 1N); 7.25 (m, 1H); 7.35 (m, 2H); 7.42 (m, 2H); 7.63 (m, 2H); 7.80 (m, 2H).

(256) UPLC-MS (AP+): 571.3 (M+Na).sup.+

Example D15

Preparation of the benzyl ester of N()-[(2RS)-2-hydroxy-3-methylselenopropanoyl]-N()-benzyloxycarbonyl-(S)-lysine (Compound 34)

(257) ##STR00047##

(258) Compound 34 is obtained using the conditions of Example D9, starting with 400 mg (2.12 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) and 862 mg (2.12 mmol; 1 eq.) of the benzyl ester of the hydrochloride of N()-benzyloxycarbonyl-(S)-lysine.

(259) The yield consists of 944 mg (76%) of the desired product in the form of a yellow oil.

(260) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 34a (R,S) and 34b (S,S): (ppm): 1.35 (m, 2H); 1.51 (m, 2H); 1.75 (m, 2H); 1.91 (m, 1H); 2.00 and 2.03 (2s, 3H); 2.89 (m, 1H); 3.07 (m, 1H); 3.17 (m, 2H); 3.37 and 3.49 (2s, 1H); 4.23 (m, 1H); 4.67 (m, 1H); 4.81 (m, 1H); 5.12-5.26 (m, 4H); 7.33-7.43 (m, 10H).

Example D16

Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-(Compound 36) D16.1. Preparation of the methyl ester of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 35)

(261) ##STR00048##

(262) Compound 35 is obtained using the conditions of Example D9, starting with 136 mg (720 mol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid (10) and 187 mg (720 mol; 1 eq.) of the methyl ester of the hydrochloride of (S)-selenomethionine.

(263) The yield consists of 180 mg (63%) of the desired product in the form of a colorless oil.

(264) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 35a (R,S) and 35b (S,S): (ppm): 2.03 (s, 3H); 2.04 and 2.07 (2s, 3H); 2.13 (m, 1H); 2.28 (m, 1H); 2.57 (t, J=7.5 Hz, 2H); 2.92 (m, 1H); 3.10 (m, 1H); 3.38 (m, 1H); 3.79 and 3.80 (2s, 3H); 4.25 (m, 1H); 4.75 (m, 1H); 7.33 and 7.37 (2d, J=8.5 Hz, 1H).

D16.2. Preparation of [(2RS)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 36)

(265) ##STR00049##

(266) Compound 36 is obtained using the conditions of Example B1, starting with 130 mg of compound 35.

(267) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (210 mL). The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (310 mL). The organic phases of this second extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(268) The yield consists of 112 mg (89%) of the desired product in the form of a yellow oil.

(269) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 36a (R,S) and 36b (S,S): (ppm): 2.05 (s, 3H); 2.08 (s, 3H); 2.17 (m, 1H); 2.33 (m, 1H); 2.63 (m, 2H); 2.92 (m, 1H); 3.10 (m, 1H); 4.26 and 4.35 (2dd, J=8.0 Hz, J=4.5 Hz, 1H); 4.75 (m, 1H); 7.43 and 7.49 (2d, J=8.0 Hz, 1H).

Example D17

Preparation of [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 36a)

D17.1. Preparation of the methyl ester of [(2R)-2-hydroxy-3-methylselenopropanoyl]-S-selenomethionine (Compound 35a)

(270) ##STR00050##

(271) Compound 35a is obtained using the conditions of Example D9, starting with 185 mg (950 mol; 1 eq.) of (R)-2-hydroxy-3-(methylseleno)propanoic acid (11) and 260 mg (950 mol; 1 eq.) of the methyl ester of the hydrochloride of (S)-selenomethionine.

(272) The yield consists of 233 mg (62%) of the desired product in the form of a pale yellow oil.

(273) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.03 (s, 3H); 2.07 (s, 3H); 2.12 (m, 1H); 2.29 (m, 1H); 2.57 (m, 2H); 2.93 (dd, J=13.0 Hz, J=7.5 Hz, 1H); 3.09 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.32 (m, 1H); 3.80 (s, 3H); 4.27 (m, 1H); 4.75 (m, 1H); 7.37 (d, J=8.0 Hz, 1H).

D17.2. Preparation of [(2R)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 36a)

(274) ##STR00051##

(275) Compound 36a is obtained using the conditions of Example B1, starting with 183 mg of compound 35a.

(276) The reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (210 mL). The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (310 mL). The organic phases are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(277) The yield consists of 178 mg (100%) of the desired product in the form of a cream colored solid.

(278) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.05 (s, 3H); 2.08 (s, 3H); 2.18 (m, 1H); 2.35 (m, 1H); 2.64 (m, 2H); 2.91 (dd, J=13.0 Hz, J=7.0 Hz, 1H); 3.09 (dd, J=13.0 Hz, J=4.0 Hz, 1H); 4.38 (dd, J=7.0 Hz, J=4.0 Hz, 1H); 4.74 (m, 1H); 7.53 (d, J=8.0 Hz, 1H).

Example D18

Preparation of the methyl ester of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 36b)

D18.1. Preparation of the methyl ester of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 35b)

(279) ##STR00052##

(280) Compound 35b is obtained using the conditions of Example D9, starting with 177 mg (949 mol; 1 eq.) of(S)-2-hydroxy-3-(methylseleno)propanoic acid (12) and 260 mg (950 mol; 1 eq.) of the methyl ester of the hydrochloride of (S)-selenomethionine.

(281) The yield consists of 236 mg (58%) of the desired product in the form of a pale yellow oil.

(282) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.03 (s, 3H); 2.04 (s, 3H); 2.12 (m, 1H); 2.29 (m, 1H); 2.57 (t, J=7.5 Hz, 2H); 2.92 (dd, J=13.0 Hz, J=8.0 Hz, 1H); 3.10 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 3.79 (s, 3H); 4.22 (dd, J=8.0 Hz, J=4.5 Hz, 1H); 4.75 (m, 1H); 7.33 (d, J=8.5 Hz, 1H).

D18.2. Preparation of [(2S)-2-hydroxy-3-methylselenopropanoyl]-(S)-selenomethionine (Compound 36b)

(283) ##STR00053##

(284) Compound 36b is obtained using the conditions of Example B1, starting with 186 mg of compound 35b.

(285) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (210 mL). The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (310 mL). The organic phases are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue is purified on a silica column (cyclohexane/ethyl acetate, then dichloromethane/methanol). The oil obtained is triturated with cyclohexane, then with pentane, dissolved in TBME and dichloromethane, then concentrated to dryness.

(286) The yield consists of 90 mg (50%) of the desired product in the form of a yellow oil.

(287) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.05 (s, 6H); 2.19 (m, 1H); 2.34 (m, 1H); 2.63 (m, 2H); 2.91 (dd, J=13.5 Hz, J=8.0 Hz, 1H); 3.12 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.26 (dd, J=8.0 Hz, J=4.5 Hz, 1H); 4.75 (m, 1H); 7.41 (d, J=8.0 Hz, 1H).

(288) 4Preparation of the Compounds E: 2-acyloxy-3-(alkylseleno)propanoic acids and corresponding esters

(289) The Compounds E are prepared either in one step by reacting the compounds B with carboxylic anhydrides, or in two steps from the compounds A by reactions with carboxylic anhydrides, followed by hydrolysis.

(290) ##STR00054##

Example E1

Preparation of 2-(acetyloxy)-3-(methylseleno)propanoic acid (Compound 37)

(291) ##STR00055##

(292) 374 mg (2 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved under nitrogen in 33 mL of dichloromethane. 817 mg (756 L; 8 mmol; 4 eq.) of acetic anhydride, then 2.5 mg (20 mol; 0.01 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is left under stirring under nitrogen and at ambient temperature for 6 h.

(293) 817 mg (756 L; 8 mmol; 4 eq.) of acetic anhydride are again added to the medium. The medium is stirred under nitrogen and at ambient temperature for 16 h.

(294) 10 mL of water are added, then the dichloromethane is eliminated from the medium by evaporation. 40 mL of a saturated NH.sub.4Cl aqueous solution are added, then the medium is extracted with ethyl acetate (340 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated. The oil obtained is purified on a silica column (cyclohexane/ethyl acetate with 1% TFA). The yield consists of 292 mg (63%) of compound 37 in the form of a yellow oil.

(295) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm): 2.15 (s, 3H); 2.20 (s, 3H); 2.97 (dd, J=13.5 Hz, J=7.5 Hz, 1H); 3.04 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 5.37 (dd, J=7.5 Hz, J=4.5 Hz, 1H); 9.90 (s, 1H).

(296) .sup.13C NMR (CDCl.sub.3, 75 MHz): (ppm): 6.2; 21.0; 25.0; 72.6; 170.7

(297) UPLC-MS (AP): 224.7 (MH.sup.+)

Example E2

Preparation of 2-(dodecanoyloxy)-3-(methylseleno)propanoic acid (Compound 38)

(298) ##STR00056##

(299) 400 mg (2.14 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved under nitrogen in 35 mL of dichloromethane. 3.344 g (8.56 mmol; 4 eq.) of lauric anhydride, then 2.6 mg (21 mol; 0.01 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 6 h.

(300) 3.344 g (8.56 mmol; 4 eq.) of lauric anhydride are added again to the medium. The medium is stirred under nitrogen and at ambient temperature for 16 h.

(301) 10 mL of water are added, then the medium is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate, then dichloromethane/methanol). The yield consists of 394 mg (47%) of compound 38 in the form of a yellow oil.

(302) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm): 0.91 (m, 311); 1.29 (m, 16H); 1.68 (m, 2H); 2.13 (s, 3H); 2.44 (t, J=7.5 Hz, 2H); 3.00 (m, 2H); 5.30 (m, 1H).

Example E3

Preparation of 2-(benzoyloxy)-3-(methylseleno)propanoic acid (Compound 40)

Example 3.1

Preparation of the methyl ester of 2-(benzoyloxy)-3-(methylseleno)propanoic acid (Compound 39)

(303) ##STR00057##

(304) 500 mg (2.46 mmol) of compound 1 are dissolved under nitrogen in 40 mL of dichloromethane. 1.136 g (4.92 mmol; 2 eq.) of benzoic anhydride, then 30 mg (246 mol; 0.1 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen at ambient temperature for 25 h.

(305) The medium is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 633 mg (81%) of compound 39 in the form of a colorless oil.

(306) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.17 (s, 3H); 3.13 (m, 2H); 3.83 (s, 3H); 5.60 (t, J=6.0 Hz, 1H); 7.50 (t, J=7.5 Hz, 2H); 7.63 (t, J=7.5 Hz, 1H); 8.13 (d, J=7.5 Hz, 2H).

E3.2: Preparation of 2-(benzoyloxy)-3-(methylseleno)propanoic acid (Compound 40)

(307) ##STR00058##

(308) Compound 40 is obtained using the conditions of Example B1, starting with 372 mg of compound 39.

(309) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (10 mL), then extracted with ethyl acetate (210 mL). The pH of the medium is adjusted to 1 by adding a 2 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (310 mL). The organic phases of this second extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(310) The yield consists of 283 mg (81%) of the desired product in the form of a white solid.

(311) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.13 (s, 3H); 2.99 (dd, J=13.5 Hz, J=6.0 Hz, 1H); 3.09 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.58 (dd, J=6.0 Hz, J=4.5 Hz, 1H); 7.52 (t, J=7.5 Hz, 2H); 7.66 (t, J=7.5 Hz, 1H); 8.15 (d, J=7.5 Hz, 2H).

Example E4

Preparation of the methyl ester of 3-(methylseleno)-2-[(3-pyridine)oxycarbonyl]propanoic acid (Compound 41)

(312) ##STR00059##

(313) 384 mg (1.85 mmol) of compound 1 are dissolved under nitrogen in 30 mL of dichloromethane. 872 mg (3.7 mmol; 2 eq.) of 3-pyridinecarboxylic anhydride, then 23 mg (185 mol; 0.1 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 18 h. The medium is filtered through fritted glass, then the filtrate is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(314) The yield consists of 490 mg (83%) of compound 41 in the form of a colorless oil.

(315) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.16 (s, 3H); 3.13 (m, 2H); 3.84 (s, 3H); 5.60 (dd, J=7.0 Hz, J=5.0 Hz, 1H); 7.47 (dd, J=8.0 Hz, J=5.0 Hz, 1H); 8.39 (d, J=8.0 Hz, 1H); 8.85 (d, J=5.0 Hz, 1H); 9.31 (s, 1H).

Example E5

Preparation of 2-[(tert-butoxycarbonyl)oxy]-3-(methylseleno)propanoic acid (Compound 43)

E5.1: Preparation of the methyl ester of 2-[(tert-butoxycarbonyl)oxy]-3-(methylseleno)propanoic acid (Compound 42)

(316) ##STR00060##

(317) 100 mg (507 mol) of compound 1 are dissolved in 5 mL of ethyl acetate. The medium is cooled to 10 C., then 103 mg (143 L; 1.01 mmol; 2 eq.) of triethylamine are added dropwise. 120 mg (533 mol; 1.05 eq.) of di-tert-butyl dicarbonate dissolved in 1 mL of ethyl acetate are added rapidly dropwise. The medium is heated at 90 C. for 48 h.

(318) The reaction medium is diluted with ethyl acetate (25 mL), then the medium is washed with a 5% citric acid aqueous solution (210 mL), then with a saturated NaCl aqueous solution (10 mL). The organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(319) The yield consists of 101 mg (63%) of compound 42 in the form of a colorless oil.

(320) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.55 (s, 9H); 2.13 (s, 3H); 2.94 (dd, J=13.5 Hz, J=7.5 Hz, 1H); 3.00 (dd, J=13.5 Hz, J=5.0 Hz, 1H); 3.82 (s, 3H); 5.17 (dd, J=7.5 Hz, J=5.0 Hz, 1H).

E5.2: Preparation of 2-[(tert-butoxycarbonyl)oxy]-3-(methylseleno)propanoic acid (Compound 43)

(321) ##STR00061##

(322) Compound 43 is obtained using the conditions of Example B1, starting with 363 mg of compound 42.

(323) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (25 mL), then extracted with ethyl acetate (220 mL). The pH of the medium is adjusted to 4 by adding a 5% citric acid aqueous solution (210 mL). The medium is extracted with ethyl acetate (525 mL). The organic phases of the second extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated.

(324) The residue is purified on a silica column (dichloromethane/methanol), then purified again on a silica column (cyclohexane/ethyl acetate).

(325) The yield consists of 39 mg (11%) of the desired product in the form of a colorless oil.

(326) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.52 (s, 9H); 2.14 (s, 3H); 2.96 (dd, J=13.5 Hz, J=7.5 Hz, 1H); 3.03 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 5.20 (dd, J=7.5 Hz, J=4.5 Hz, 1H); 10.04 (s, 1H).

Example E6

Preparation of the methyl ester of (2RS)[N-(tert-butoxycarbonyl)-S-methionyl]-3-methylselenopropanoic acid (Compound 44)

(327) ##STR00062##

(328) 500 mg (2.0 mmol) of BOC(S)-methionine are dissolved in 25 mL of dichloromethane. 417 mg (2.0 mmol; 1 eq.) of N,N-dicyclohexylcarbodiimide are added. The medium is stirred for 10 min at ambient temperature, then 416 mg (2.0 mmol; 1 eq.) of compound 1 and 25 mg (200 mol; 0.1 eq.) of 4-dimethylaminopyridine are added. The mixture is stirred at ambient temperature for 16 h.

(329) The medium is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(330) The yield consists of 709 mg (78%) of compound 44 in the form of a colorless oil.

(331) .sup.1H NMR (CDCl.sub.3, 400 MHz) 50/50 mixture of 2 diastereoisomers 44a (R,S) and 44b (S,S): (ppm)=1.48 (s, 9H); 2.04 (m, 1H); 2.12 and 2.14 (2s, 3H); 2.15 and 2.16 (2s, 3H); 2.26 (m, 1H); 2.65 (m, 2H); 2.99 (m, 2H); 3.80 and 3.81 (2s, 3H); 4.58 (m, 1H); 5.17 (m, 1H); 5.39 (m, 1H).

(332) UPLC-MS (AP+): 451.9 (M+Na).sup.+

Example E7

Preparation of the methyl ester of 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid (Compound 45) and 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid (Compound 46)

E7.1. Preparation of the methyl ester of 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid (Compound 45)

(333) ##STR00063##

(334) 128 mg (522 mol) of 2-(acetyloxy)-3-(methylseleno)propanoic acid (37) are dissolved under nitrogen in 5 mL of dichloromethane. 108.7 mg (522 mol) of N,N-dicyclohexylcarbodiimide are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 10 min. 116 mg (522 mol) of methyl 2-hydroxy-4-(methylseleno)butanoate (EP1778706, prepared in a similar manner to Example 9 using methanol instead of ethanol) in solution in 5 mL of dichloromethane, then 6.4 mg (52 mol; 0.01 eq.) of 4-dimethylaminopyridine are added. The medium is stirred at ambient temperature for 16 h.

(335) The medium is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(336) The yield consists of 189 mg (77%) of compound 45 in the form of a colorless oil.

(337) .sup.1H NMR (CDCl.sub.3, 400 MHz) mixture of 2 diastereoisomers: (ppm)=2.03 and 2.04 (2s, 3H); 2.15 and 2.16 (2s, 3H); 2.20 (s, 3H); 2.23-2.3 (m, 2H); 2.54-2.7 (m, 2H); 2.95-3.02 (m, 1H); 3.11 (dd, J=13.5 Hz, J=4.0 Hz, 1H); 3.79 and 3.80 (2s, 3H); 5.15 to 5.45 (m, 2H).

(338) UPLC-MS (AP+): 442.9 (M+Na).sup.+

E7.2. Preparation of 4-methylseleno-2-(2-acetyloxy-3-methylselenopropanoyl)butyric acid (Compound 46) by hydrolysis of the methyl ester

(339) ##STR00064##

(340) Compound 46 is obtained using the conditions of Example B1, starting with 180 mg of compound 45.

(341) After 16 h of stirring at ambient temperature, the reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (25 mL). The aqueous phase is lyophilized. The lyophilizate is redissolved with a 4 N hydrochloric acid solution in dioxane. The medium is stirred for 10 min and then concentrated to dryness. The concentrate is redissolved in 10 mL of water, then the solution is lyophilized.

(342) The yield consists of 138 mg (71%) of the desired product containing 2 LiCl in the form of a yellow sticky solid.

(343) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.00 (s, 3H); 2.05 (s, 3H); 2.07-2.24 (m, 2H); 2.65 (m, 2H); 2.89 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 3.00 (dd, J=13.0 Hz, J=4.5 Hz, 1H); 4.36 (dd, J=8.0 Hz, J=4.0 Hz, 1H); 4.50 (dd, J=6.5 Hz, J=4.5 Hz, 1H).

Example E8

Preparation of 3-methylseleno-2-(2-hydroxy-4-methylselenobutanoyl)propanoic acid (Compound 48)

E8.1: Preparation of the methyl ester of 3-methylseleno-2-(2-hydroxy-4-methylselenobutanoyl)propanoic acid (Compound 47)

(344) ##STR00065##

(345) Compound 47 is obtained using the conditions of Example E7, starting with 89 mg of 2-acetyloxymethylselenobutyric acid (EP1778706, Example 11) and 72 mg of compound 1.

(346) The medium is filtered through fritted glass, then the filtrate is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate).

(347) The yield consists of 97 mg (60%) of the desired product in the form of a colorless oil.

(348) .sup.1H NMR (CDCl.sub.3, 400 MHz) mixture of diastereoisomers: (ppm)=2.05 and 2.06 (2s, 3H); 2.11 and 2.14 (2s, 3H); 2.18 and 2.19 (2s, 3H); 2.27-2.37 (m, 2H); 2.60-2.80 (m, 2H); 2.92-3.07 (m, 2H); 3.80 and 3.81 (2s, 3H); 5.24 (dd, J=12.5 Hz, J=6.0 Hz, 1H); 5.34 and 5.45 (2dd, J=8.0 Hz, J=4.5 Hz, 1H).

(349) UPLC-MS (AP+): 443.0 (M+Na).sup.+

E8.2: Preparation of 3-methylseleno-2-(2-hydroxy-4-methylselenobutanoyl)propanoic acid (Compound 48)

(350) ##STR00066##

(351) Compound 48 is obtained using the conditions of Example E7.2, starting with 79 mg of compound 47.

(352) The yield consists of 69 mg (84%) of the desired product containing 2 LiCl in the form of a colorless sticky solid.

(353) .sup.1H NMR (D.sub.2O, 400 MHz): (ppm)=2.04 (m, 3H); 2.09-2.14 (m, 3H); 2.20-2.27 (m, 2H); 2.64-2.74 (m, 2H); 2.93 (dd, J=13.0 Hz, J=6.5 Hz, 1H); 3.04 (dd, J=13.5 Hz, J=4.5 Hz, 1H); 4.55 (dd, J=6.5 Hz, J=4.5 Hz, 1H); 5.05 (m, 1H).

Example E9

Preparation of 2-(pentanoyloxy)-3-(methylseleno)propanoic acid (Compound 49)

(354) ##STR00067##

(355) 407 mg (1.98 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved under nitrogen in 33 mL of dichloromethane. 752 mg (817 L; 3.96 mmol; 2 eq.) of valeric anhydride, then 2.4 mg (19.8 mol; 0.01 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 24 h.

(356) 752 mg (817 L; 3.96 mmol; 2 eq.) of valeric anhydride, then 2.4 mg (19.8 mol; 0.01 eq.) of 4-dimethylaminopyridine are again added to the medium. The medium is stirred under nitrogen and at ambient temperature for 48 h. 10 mL of water are added, then the dichloromethane is eliminated from the medium by evaporation. 25 mL of a saturated NH.sub.4C aqueous solution are added, then the medium is extracted with ethyl acetate (310 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated. The oil obtained is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 260 mg (48%) of compound 49 in the form of a colorless oil.

(357) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=0.96 (t, J=7.4 Hz, 3H); 1.42 (m, 2H); 1.69 (m, 2H); 2.15 (s, 3H); 2.46 (td, J=1.9 Hz, J=7.4 Hz, 2H); 3.01 (m, 2H); 5.36 (dd, J=4.3 Hz, J=7.6 Hz, 1H).

(358) UPLC-MS (AP): 267.3 (MH.sup.+)

Example E10

Preparation of 2-(nonanoyloxy)-3-(methylseleno)propanoic acid (Compound 50)

(359) ##STR00068##

(360) 260 mg (1.39 mmol) of 2-hydroxy-3-(methylseleno)propanoic acid (10) are dissolved under nitrogen in 23 mL of dichloromethane. 874 mg (963 L; 2.78 mmol; 2 eq.) of nonanoic anhydride, then 1.7 mg (13.9 mol; 0.01 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 24 h.

(361) 874 mg (963 L; 2.78 mmol; 2 eq.) of nonanoic anhydride, then 1.7 mg (13.9 mol; 0.01 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 24 h.

(362) 10 mL of water are added, then the dichloromethane is eliminated from the medium by evaporation. 25 mL of a saturated NH.sub.4Cl aqueous solution are added, then the medium is extracted with ethyl acetate (310 mL). The organic phases are combined, dried with Na.sub.2SO.sub.4, filtered and concentrated. The oil obtained is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 116 mg (25%) of compound 50 in the form of a colorless oil.

(363) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=0.91 (t, J=6.9 Hz, 3H); 1.34 (m, 10H); 1.70 (m, 2H); 2.14 (s, 3H); 2.45 (td, J=1.7 Hz, J=7.4 Hz, 2H); 3.01 (m, 2H); 5.37 (dd, J=4.3 Hz, J=7.6 Hz, 1H).

(364) UPLC-MS (AP): 322.9 (MH.sup.+)

Example E11.1

Preparation of the methyl ester of 2-(linoleoyloxy)-3-(methylseleno)propanoic acid (Compound 51)

(365) ##STR00069##

(366) 818 mg (2.83 mmol) of linoleic acid are dissolved in 35 mL of dichloromethane. 589 mg (2.83 mmol; 1 eq.) of N,N-dicyclohexylcarbodiimide are added. The medium is stirred for 10 min at ambient temperature, then 575 mg (2.83 mmol; 1 eq.) of compound 1 and 35 mg (283 mol; 0.1 eq.) of 4-dimethylaminopyridine are added. The medium is stirred at ambient temperature for 48 h.

(367) The medium is filtered through fritted glass, then the filtrate is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 801 mg (60%) of compound 51 in the form of a colorless oil.

(368) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=0.92 (t, J=6.9 Hz, 3H); 1.34 (m, 14H); 1.70 (m, 2H); 2.08 (m, 4H); 2.12 (s, 3H); 2.45 (td, J=2.8 Hz, J=7.4 Hz, 2H); 2.8 (t, J=6.6 Hz, 2H); 2.97 (m, 2H); 3.8 (s, 3H); 5.38 (m, 5H).

(369) UPLC-MS (AP+): 461.2 (M+H.sup.+)

Example E11.2

Preparation of 2-(linoleoyloxy)-3-(methylseleno)propanoic acid (Compound 52)

(370) ##STR00070##

(371) 459 mg (1.59 mmol) of linoleic acid are dissolved in 19.8 mL of dichloromethane. 331 mg (2.83 mmol; 1 eq.) of N,N-dicyclohexylcarbodiimide are added. The medium is stirred for 30 min at ambient temperature, then 300 mg (1.59 mmol; 1 eq.) of 2-hydroxy-3-(methylseleno)propanoic acid and 20 mg (159 mol; 0.1 eq.) of 4-dimethylaminopyridine are added. The medium is stirred at ambient temperature for 24 h.

(372) The medium is filtered through fritted glass, then the filtrate is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate, then dichloromethane/methanol). The yield consists of 198 mg (26%) of compound 52 in the form of a colorless oil.

(373) .sup.1H NMR (CDCl.sub.3, 300 MHz): (ppm)=0.93 (t, J=6.8 Hz, 3H); 1.35 (m, 14H); 1.70 (m, 2H); 2.1 (m, 4H); 2.15 (s, 3H); 2.45 (m, 2H); 2.8 (t, J=5.9 Hz, 2H); 3.0 (m, 2H); 5.4 (m, 5H).

(374) UPLC-MS (AP): 444.7 (M+H.sup.+)

Example E12

Preparation of the methyl ester of 2-[(1H-imidazol-4-ylcarbonyl)oxy]-3-(methylseleno)propanoic acid (Compound 53)

(375) ##STR00071##

(376) 438 mg (3.83 mmol) of 4-imidazolecarboxylic acid are suspended in 20 mL of dichloromethane and 30 L of N,N-dimethylformamide (383 mol; 0.1 eq.). The medium is cooled to 0 C., then 503 L (5.74 mmol; 1.5 eq.) of oxalyl chloride are added. The medium is cooled to 0 C. for 15 min, then stirred for 2 h at ambient temperature. The medium is concentrated to dryness.

(377) The concentrate is redissolved in 20 mL of dichloromethane. The medium is cooled to 0 C., then 1.26 mL (7.66 mmol; 2 eq.) of N,N-diisopropylethylamine are added. 794 mg (3.83 mmol; 1 eq.) of compound 1 are added dropwise. The medium is stirred at ambient temperature for 48 h.

(378) The medium is diluted with 50 mL of ethyl acetate, then washed with 20 mL of water. The aqueous phase is extracted with 20 mL of ethyl acetate. The organic phases are combined, washed with 20 mL of a saturated NaCl aqueous solution. The organic phase is dried with Na.sub.2SO.sub.4, filtered and concentrated. The residue is purified on a silica column (cyclohexane/ethyl acetate, then dichloromethane/methanol). The yield consists of 806 mg (70%) of compound 53 in the form of a pale yellow solid.

(379) .sup.1H NMR (DMSO, 400 MHz): (ppm)=2.12 (s, 3H); 3.03 (m, 21H); 3.70 (s, 3H); 5.42 (dd, J=4.6 Hz, J=7.3 Hz, 1H); 7.84 (s, 1H); 7.88 (s, 1H); 12.74 (br s, 1H).

(380) UPLC-MS (AP+): 314.7 (M+Na.sup.+)

Example E13.1

Preparation of the methyl ester of 2-(pivaloyloxy)-3-(methylseleno)propanoic acid (Compound 54)

(381) ##STR00072##

(382) 400 mg (1.99 mmol) of compound 1 are dissolved under nitrogen in 50 mL of dichloromethane. 1.5 g (7.95 mmol; 4 eq.) of trimethylacetic anhydride, then 24 mg (199 mol; 0.1 eq.) of 4-dimethylaminopyridine are added to the medium. The medium is stirred under nitrogen and at ambient temperature for 48 h. 748 mg (3.98 mmol; 2 eq.) of trimethylacetic anhydride are again added to the medium. The medium is stirred under nitrogen and at ambient temperature for 96 h.

(383) The medium is concentrated to dryness. The residue is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 484 mg (83%) of compound 54 in the form of a colorless oil.

(384) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.29 (s, 9H); 2.13 (s, 3H); 2.99 (m, 2H); 3.79 (s, 3H); 5.31 (dd, J=4.6 Hz, J=7.5 Hz, 1H).

(385) UPLC-MS (AP+): 281.9 (M+H.sup.+)

Example E13.2

Preparation of 2-(pivaloyloxy)-3-(methylseleno)propanoic acid (Compound 55)

(386) ##STR00073##

(387) 240 mg (0.83 mmol) of compound 54 are dissolved in 12 mL of THF. 830 L (0.83 mmol; 1 equiv.) of a 1M lithium hydroxide aqueous solution are added, the solution is stirred at ambient temperature for 16 h. 41 L (41 mol, 0.05 equiv.) of a 1M lithium hydroxide aqueous solution are added, the solution is stirred at ambient temperature for 24 h.

(388) The reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (320 mL). The pH of the medium is adjusted to 4 by adding a 1 M hydrochloric acid aqueous solution. The medium is extracted with ethyl acetate (320 mL). The organic phases of this extraction are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated. The yield consists of 151 mg (66%) of the desired product 55 in the form of a colorless oil.

(389) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=1.29 (s, 9H); 2.15 (s, 3H); 3.03 (m, 2H); 5.32 (dd, J=7.7 Hz, J=4.3 Hz, 1H).

(390) UPLC-MS (AP): 266.6 (M+H.sup.+)

Example E14

Preparation of the methyl ester of 2-(3-chloropropanoyloxy)-3-(methylseleno)propanoic acid (Compound 56)

(391) ##STR00074##

(392) 400 mg (1.99 mmol) of compound 1 are suspended under nitrogen in 6 mL of acetone. 324 mg (3.98 mmol; 2 eq.) of pyridine are added to the medium. The medium is cooled to 10 C., then 390 L (3.98 mmol; 2 eq.) of 3-chloropropionyl chloride are added to the medium at 10 OC. The medium is stirred at ambient temperature for 64 h.

(393) The reaction medium is diluted in water (20 mL), then extracted with ethyl acetate (220 mL). The organic phases are combined, washed with 220 mL of a 1M NaHCO.sub.3 aqueous solution and 10 mL of a saturated NaCl aqueous solution. The organic phase is dried with Na.sub.2SO.sub.4, filtered and concentrated. The residue is purified on a silica column (cyclohexane/ethyl acetate). The yield consists of 301 mg (51%) of compound 56 in the form of a colorless oil.

(394) .sup.1H NMR (CDCl.sub.3, 400 MHz): (ppm)=2.13 (s, 3H); 2.97 (m, 4H); 3.81 (s, 3H); 3.82 (t, J=6.6 Hz, 2H); 5.4 (dd, J=4.6 Hz, J=7.5 Hz, 1H).

(395) Overview Table of the Preparation Examples of the Novel Compounds

(396) TABLE-US-00001 Com- Ex. pound Structure R1 R2 X A1 1 CH.sub.3 H OCH.sub.3 A2 1 CH.sub.3 H OCH.sub.3 A3 2 CH.sub.3 H OCH.sub.3 A4 3 CH.sub.3 H OCH.sub.3 A5 4 CH.sub.3 H OCH.sub.2CH.sub.3 A6 5 0 CH.sub.3 H OC(CH.sub.3).sub.3 A7 6 CH.sub.3 H OCH.sub.2C.sub.6H.sub.5 A8 7 CH.sub.2CH.sub.3 H OCH.sub.3 A9 8 CH.sub.2CH(CH.sub.3).sub.2 H OCH.sub.3 A10 4 CH.sub.3 H OCH.sub.2CH.sub.3 A11 9 CH.sub.3 H OCH(CH.sub.3).sub.2 B1 10 CH.sub.3 H OH B2 10 CH.sub.3 H OH B3 11 CH.sub.3 H OH B4 12 CH.sub.3 H OH B5 12 0 CH.sub.3 H OH B6 13 CH.sub.2CH.sub.3 H OH B7 14 CH.sub.2CH(CH.sub.3).sub.2 H OH C1 15 CH.sub.3 H ONH.sub.2(C.sub.6H.sub.11).sub.2 C2 16 CH.sub.3 H Na salt C3 17 CH.sub.3 H Mg salt C4 18 CH.sub.3 H Zn salt C5 19 CH.sub.3 H Ca salt D1 20 CH.sub.3 H NH.sub.2 D2 21 CH.sub.3 H NHC.sub.3H.sub.5 D3 22 00 CH.sub.3 H NH(CH.sub.2).sub.2N(CH.sub.3).sub.2 D4 23 01 CH.sub.3 H N(CH.sub.2).sub.4 D5 24 02 CH.sub.3 H N(CH.sub.2).sub.3 D6 25 03 CH.sub.3 H N(CH.sub.2).sub.4O D7 26 04 CH.sub.3 H N(C.sub.2H.sub.5).sub.2 D8 27 05 CH.sub.2 H NH(CH.sub.2).sub.2OH D9.1 28 06 CH.sub.3 H NHAlaOC(CH.sub.3).sub.3 D9.2 29 07 CH.sub.3 H NHAlaOH D10.1 30 08 CH.sub.3 H NCHMetOCH.sub.3 D10.2 31 09 CH.sub.3 H NHMetOH D11.1 30a 0 CH.sub.3 H NHMetOCH.sub.3 D11.2 31a CH.sub.3 H NHMetOH D12.1 30b CH.sub.3 H NHMetOCH.sub.3 D12.2 31b CH.sub.3 H NHMetOH D13 32 CH.sub.3 H NHLys(NHBoc)OCH.sub.4 D14 33 CH.sub.3 H NHLys(NHFmoc)OCH.sub.3 D15 34 CH.sub.3 H NHLys(NHCbz) OCH.sub.2C.sub.6H.sub.5 D16.1 35 CH.sub.3 H NHSeMetOCH.sub.3 D16.2 36 CH.sub.3 H NHSeMetOH D17.1 35a CH.sub.3 H NHSeMetOCH.sub.3 D17.2 36a 0 CH.sub.3 H NHSeMetOH D18.1 35b CH.sub.3 H NHSetMetOCH.sub.3 D18.2 36b CH.sub.3 H NHSeMetOH E1 37 CH.sub.3 C(O)CH.sub.3 OH E2 38 CH.sub.3 C(O)(CH.sub.2).sub.10CH.sub.3 OH E3.1 39 CH.sub.3 C(O)C.sub.6H.sub.5 OCH.sub.3 E3.2 40 CH.sub.3 C(O)C.sub.6H.sub.5 OH E4 41 CH.sub.3 C(O)C.sub.6H.sub.4N OCH.sub.3 E5.1 42 CH.sub.3 C(O)OC(CH.sub.3).sub.3 OCH.sub.3 E5.2 43 CH.sub.3 C(O)OC(CH.sub.3).sub.3 OH E6 44 0 CH.sub.3 MetNHBoc OCH.sub.3 E7.1 45 CH.sub.3 C(O)CH.sub.3 OSeMetOCH.sub.3 E7.2 46 CH.sub.3 H OSeMetOH E8.1 47 CH.sub.3 C(O)CH.sub.3 OSeMetOCH.sub.3 E8.2 48 CH.sub.3 H OSeMetOH E9 49 CH.sub.3 C(O)C.sub.4H.sub.7 OH E10 50 CH.sub.3 C(O)C.sub.4H.sub.9 OH E11.1 51 CH.sub.3 C(O)C.sub.17H.sub.21 OCH.sub.3 E11.2 52 CH.sub.3 C(O)C.sub.17H.sub.21 OH E12 53 CH.sub.3 C(O)C.sub.3H.sub.3N.sub.2 OCH.sub.3 E13.1 54 0 CH.sub.3 C(O)C(CH.sub.3).sub.2 OCH.sub.3 E13.2 55 CH.sub.3 C(O)C(CH.sub.3).sub.5 OH E14 56 CH.sub.3 C(O)(CH.sub.2).sub.2Cl OCH.sub.3
II. Examples Describing the Antitumor Activity of the Compounds According to the Invention
II.1. Cell Lines

(397) Eight cell lines originating from different types of cancer were used: PC3 and DU145 (prostate), HT-29 and LS-174T (colon), Hep G2 (liver), MCF-7 (breast), MIA PaCa-2 and PANC-1 (pancreas).

(398) The characteristics of each cell line are summarized in Table I.

(399) TABLE-US-00002 TABLE I Characteristics of the cell lines used Organ Cell line Description Prostate PC3 Adherent, human prostate adenocarcinoma grade IV derived from osseous metastasis DU145 Adherent, human prostate carcinoma Colon HT-29 Adherent, human adenocarcinoma (44-year old Caucasian male) LS174T Adherent, human adenocarcinoma (58-year old Caucasian woman) Liver Hep G2 Adherent, hepatocellular carcinoma (15-year old Caucasian) Breast MCF-7 Adherent, human adenocarcinoma (69-year old Caucasian woman) Pancreas PANC-1 Adherent, carcinoma (56-year old Caucasian man) MIA Adherent, carcinoma (65-year old Caucasian PaCa-2 man)
II.2. Culture Media

(400) The cells are cultured in the specific culture medium described in Table II, at 37 C., 5% CO.sub.2, according to the operating procedures that are well known to the person skilled in the art.

(401) TABLE-US-00003 TABLE II Composition of the culture media Organ Cell line Culture medium Prostate PC3 RPMI 1640 + 10% FBS DU145 RPMI 1640 + 10% FBS Colon HT-29 McCoy's 5a + 10% FBS + 0.5 mM ultraglutamine LS174T EMEM + 10% FBS + 2 mM ultraglutamine + 1 mM sodium pyruvate + 0.1 mM nonessential amino acids Breast MCF-7 EMEM + 10% FBS + 2 mM ultraglutamine + 1 mM sodium pyruvate + 0.1 mM nonessential amino acids + 10 nM -estradiol Pancreas PANC-1 RPMI + 10% FBS MIA DMEM + 10% FBS PaCa-2 Liver Hep G2 EMEM + 10% of complemented FBS + 0.1 mM nonessential amino acids + 2 mM ultraglutamine + penicillin/streptomycin
II.3. Evaluation of the Cytotoxicity of the Compounds According to the Invention

(402) After thawing and amplification of the cancer cells in the appropriate culture medium (described in 1.2.), 96-well plates are inoculated with these cells and incubated or not incubated (controls) in their respective culture medium, in the presence of the compounds to be tested at 10, 50, 100, 250 and 500 M.

(403) After 96 h of incubation, each 96-well plate is analyzed in order to measure the viability of the cells using a colorimetric test with WST-1.

(404) II.4 Examples

(405) F1. Antitumor Activity of Compound 4 (See Example A5)

(406) The cytotoxicity results obtained with compound 4 on two cell lines are presented in FIG. 2. These results show that the viability of the cells decreases as the concentration of compound 4 increases, the concentration of compound 4 decreases by 50%, the cell viability being equal to 481 and 335 M for the cells DU145 and LS174T, respectively.

(407) F2. Antitumor Activity of Compound 10 (See Example B1)

(408) The cytotoxicity results obtained with compound 10 on three cell lines are presented in FIG. 3. These results show that the viability of the cells decreases as the concentration of compound 10 increases, the concentration of compound 10 decreases by 50%, the cell viability being equal to 392, 430 and 328 M for the cells DU145, LS174T and HT-29, respectively.

(409) F3. Antitumor Activity of Compound 38 (See Example E2)

(410) The cytotoxicity results obtained with compound 38 on eight cell lines are presented in FIGS. 4a and 4b. These results show that the viability of the cells decreases as the concentration of compound 38 increases. The concentration of compound 38 which decreases by 50%, the cell viability is given in Table III for each cell line.

(411) TABLE-US-00004 TABLE III Concentration of compound 38 decreasing by 50% the cell viability after 96 hours of treatment Compound 38 Cell line (M) PC3 198 DU145 169 PANC-1 253 MIA PaCa-2 308 HT-29 166 LS174T 113 Hep G2 263 MCF-7 215
III. Examples Describing the Compositions of the Compounds According to the Invention

Example G1

Compositions Containing Compound 10

(412) Capsules having the following composition were prepared:

(413) TABLE-US-00005 Compound 10 2-Hydroxy-3-methylselenopropanoic 0.40 mg (in Se eq.) acid Excipients* and envelope** a 1000 mg capsule (*cornstarch, lactose, magnesium stearate, flavor, **gelatin, titanium dioxide, dyes)

Example G2

Compositions Containing Compound 38

(414) Capsules having the following composition were prepared:

(415) TABLE-US-00006 Compound 38 2-(Dodecanoyloxy)-3-(methylseleno) 0.40 mg (in Se eq.) propanoic acid Excipients* and envelope** a 1000 mg capsule (*cornstarch, lactose, magnesium stearate, flavor, **gelatin, titanium dioxide, dyes)

Example G3

Compositions Containing Compound 10 and Compound 38

(416) Capsules having the following compositions were prepared:

(417) TABLE-US-00007 Compound 10 2-Hydroxy-3-methylselenopropanoic 0.10 mg (in Se eq.) acid Compound 38 2-(Dodecanoyloxy)-3- 0.40 mg (in Se eq.) (methylseleno)propanoic acid Excipients* and envelope** a 1000 mg capsule (*cornstarch, lactose, magnesium stearate, flavor, **gelatin, titanium dioxide, dyes)