HU217960B - Compounds with a fused bicyclic ring containing one nitrogen and sulfur or oxygen atoms and process for producing them - Google Patents

Abstract

SUMMARY OF THE INVENTION The present invention relates to compounds of formula III wherein X is oxygen or sulfur; m = 0 or 1; n = 1 or 2; Y is a methylene group or an oxygen atom, with the proviso that when Y is oxygen then m = 1; R 3 is hydrogen or alkyl; and with the further limitation that when X is oxygen, n = 1, and Y is methyl, then m = 1 - and its salts, and a process for their production and their carboxyl-protected derivatives. The compounds of the present invention are intermediates for the preparation of angiotensin converting enzyme and compounds that inhibit the function of neutral endopeptidases. ŕ

Description

The present invention relates to compounds of formula (III) containing a fused bicyclic ring and to processes for their preparation and their preparation on carbonyl-protected derivatives. The compounds of formula (III) are useful as intermediates in the preparation of compounds containing a fused bicyclic ring described in Hungarian Patent Application No. 216,791, which also act as inhibitors of the action of angiotensin converting enzyme (ACE) and, in some cases, of neutral endopeptidases (NCPs). They are useful in the manufacture of a medicament for the treatment of ACE inhibitors, for example for the treatment of hypertensive disorders.

In the formula (III), X is oxygen or sulfur; n is 1 or 2;

m is 0 or 1;

Y is methylene (-CH ₂ -) or oxygen, with the proviso that Y can be oxygen only when m is 1;

R ₃ is hydrogen or C 1-7 alkyl; and further provided that when X is oxygen, n is 1 and Y is methylene, then m is 1.

The term C 1-7 alkyl as used herein, refers to a group derived from straight or branched chain hydrocarbons. Carboxy protecting groups are also known from T. W. Greene's "Protective Groups in Organic Synthesis" (John Wiley & Sons, Wiley Interscience, N. Y., 1967).

The compounds of formula (III) may be prepared in a variety of ways. These methods are described below.

In the preparation of compounds of formula (III) wherein Y is methylene, an amino acid of formula (XXII) protected at its amino group is first coupled to a dipeptide of formula (XXIV) by an amino acid ester of formula (XXIII). In the formulas P | is an amino protecting group such as benzyloxycarbonyl or tert-butoxycarbonyl, or a group commonly referred to as the protecting group with a protected nitrogen atom, such as a phthalimido group; P ₂ is a protecting group for a hydroxy or mercapto group; and R ₃ is an easily removable esterification group protecting the carboxy group. Of the protecting groups corresponding to P ₂ , when X is sulfur, acyl groups such as acetyl or benzoyl, but more preferably acetyl, are preferred, while when X is oxygen, P _{2 is} preferably acyl, tetrahydropyranyl, trityl or sterically hindered silyl groups. but especially triphenylmethyl and tert-butyldimethylsilyl. The coupling reaction is conveniently carried out with a condensing agent such as [(benzotriaziazol-1-yl) oxy] tris (dimethylamino) phosphonium [hexafluorophosphate], 1- [3- (dimethylamino) propyl] -3-. ethyl carbodiimide or [(methylsulfonyl) oxybenzotriazole].

Further conversion of intermediates of formula (XXIV) is carried out by the subsequent step of selectively removing the P ₂ protecting group, for example in methanol, with sodium methylate when P ₂ is acetyl or benzoyl, or with an acid such as p-toluenesulfonic acid, P _{2 represents an} acetyl, benzoyl, trityl, tetrahydropyranyl or tert-butyldimethylsilyl group and the resulting product is subjected to an acid catalyzed cyclization to give a compound of formula XXV. As the acid catalyst, it is preferable to use a strong acid such as trifluoroacetic acid or p-toluenesulfonic acid, or a commercially available polystyrene sulfonate ion exchange resin such as Amberlyst (R) 15 ion exchange resin and the solvent may be an aprotic solvent such as methylene chloride.

After deprotection of P ₂ , but prior to cyclization, an intermediate of formula (XXIV) wherein X is oxygen can be converted to the corresponding intermediate of formula (XXIV) wherein X is sulfur. There are several ways of such conversion, which is conveniently carried out by reacting the compound of formula (XXIV) with triphenylphosphine, diisopropyl azodicarboxylate and thio-S-acetic acid after deprotection of the P ₂ protecting group. the thioacetate formed is hydrolyzed with sodium methylate in methanol and finally the hydrolysis product, mercaptan, is cyclized as described above.

Alternatively Method III variant yielded (XXIV) with intermediate of formula may be converted after the P ₂ protecting group is removed in a known manner a compound of formula (XXVI) wherein L is a leaving group such as methanesulfonyloxy or p-toluenesulfonyloxy group , or iodine or bromine. In a specific example, reaction of a compound of Formula XXIV with methanesulfonyl chloride after cleavage of the P ₂ protecting group affords a compound of Formula XXVI wherein L is methanesulfonyloxy.

The compounds of formula (XXVI) are reacted with cesium thioacetate to give the corresponding thioacetic acid derivative which can be converted to the desired mercaptan in methanol with sodium methylate, the final step being the cyclization discussed above.

Compounds of formula (XXIV) wherein X is oxygen, when used in a solvent such as dichloromethane or chloroform with a strong acid such as trifluoroacetic acid or p-toluenesulfonic acid or commercially available polystyrene sulfonate ion exchange resins For example, by reaction with an ion exchange resin such as Amberlyst (R) 15, compounds of formula (XXV) can be prepared directly.

The next step is now to remove the amino protecting group from molecule XXV 2

This may be done, for example, with hydrazine hydrate when P, together with the nitrogen atom, is a phthalimido group; iodomethylsilane or palladium on carbon in the presence of ammonium formate or hydrogen gas when P 1 is (benzyloxy) carbonyl; and in dioxane with hydrochloric acid or another strong acid, when P is tert-butoxycarbonyl, to give the expected fused bicyclic compound of formula (III).

Alternatively, compounds of formula (III) wherein Y is methylene may also be prepared by the first step of protecting an amino acid of formula (XXII) with a hydroxy amino acid ester of formula (XXVII) coupling to a dipeptide of formula (XXVIII) wherein P [and P ₂ are as defined above], preferably a condensing agent such as [(methylsulfonyl) oxy] benzotriazole or 1- [3- (dimethylamino) ) -propyl] -3-ethylcarbodiimide. The hydroxy compound (XXVIII) thus formed is first oxidized to an aldehyde (XXIX) by reaction with oxalyl dichloride and dimethylsulfoxide and then in an aprotic solvent to form an aldehyde of formula (XXIX). ₂ protecting groups, and finally with a suitable acid catalyst, this has already been discussed in detail above - forcing the cyclization to give the corresponding intermediate of formula (XXV).

The starting compounds of formula (XXIII) wherein m is 1 can be prepared by selectively protecting the amino group of ε-hydroxy-L-norleucine to form the corresponding compound of formula (XXX), wherein P _{3 represents an} amino protecting group, for example a phthalimido group with the nitrogen atom, and the carboxy group of this compound of formula XXX is also protected with a protecting group corresponding to R ₃ , such as methyl, which is preferably introduced into the molecule in the presence of a base. iodide or in a methanolic medium with a strong acid. In the next step, the resulting ester is oxidized to an aldehyde of formula (XXXI) and then reacted with an ortho-acetic acid ester of formula (XXXII) with a catalytic amount of acid and the corresponding alcohol - an alcohol of the formula OH in the presence of an alkyl group of formula XXXII. The reaction results in the formation of a compound of formula XXXIII. Cleavage of the protecting group P ₃ , for example with hydrazine hydrate, when P ₃ together with the nitrogen atom is a phthalimido group, results in the corresponding compound of formula XXIII, wherein m is 1.

Starting compounds of formula XXIII wherein m is zero are derived from an amino acid derivative of formula XXXIV which is protected on its amino group by a glutamic acid γ-benzyl ester, wherein P ₃ is an amino protecting group such as tert-butoxy. -carbonyl group or, together with the nitrogen atom, phthalimido group

- we can produce it. The protected glutamic acid ester of formula (XXXIV) is first protected at its free carboxy group as in the case discussed above for hydroxy norleucine, and then the compound of formula (XXXV) is obtained.

wherein R ₃ is lower alkyl

- hydrogenolysis, i.e. cleavage of the benzyl group, to give a compound of formula XXXVI. Selective reduction of compounds of formula (XXXVI), which may be accomplished first by ethanethiol, 1- [3- (dimethylamino) propyl] -3-ethylcarbodiimide and 4- (dimethylamino) pyridine, followed by reaction with triethylsilane in acetonitrile in the presence of palladium on charcoal to give the corresponding aldehyde XXXVII, from which the aldehyde can be reacted with an orthoic acid ester of formula XXXII and then deprotected P ₃ a desired starting compound of formula XXIII wherein m is zero.

The hydroxy amino acid esters of formula (XXVII) which can be used as starting materials in the synthesis of the compounds of the invention can be prepared by first adding diethyl [2- (acetylamino) malonate] to a suspension of sodium hydride under stirring, deprotonating, then deprotonating the intermediate with a (haloalkyl) acetate (XXXVIII)

reacting a halogen atom of a bromine, iodine or chlorine compound to form a compound of formula (XXXIX). The hydrolysis of the esters of formula (XXXIX) by warming with sodium hydroxide followed by heating the acid after acidification yields the corresponding hydroxy amino acid derivative of formula (XL), which is hydrolyzed with a suitable hydrolyzing enzyme such as acylase isolated from porcine kidney. ) to give the hydroxy amino acid.

The final step is the conversion of the resulting hydroxy amino acid of formula (XLI) to an ester of formula (XXVII), which may be accomplished by any of the conventional methods. For example, the appropriate hydroxy amino acid of formula (XLI) may be reacted with chlorotrimethylsilane in methanol to give the desired compound of formula (XXVII) as the methyl ester as its hydrochloride salt.

The starting compounds of formula (XXII) can be prepared as follows.

For compounds of Formula XXII wherein X is O, an alpha-amino acid of Formula XLII may be initiated and the protecting groups P ₁ and P ₂ introduced into the molecule. In one embodiment, a compound of formula (XLII) is first reacted with N-ethoxycarbonylphthalimide in the presence of sodium carbonate followed by trityl chloride in the presence of triethylamine. This gives a starting compound of formula XXII wherein X is O and P, together with the nitrogen atom, is a phthalimido group, P ₂

HU 217 960 jelent represents a trityl group. Alternatively, an acid of formula (XLII) may first be reacted with N - [(benzyloxy) carbonyl] succinimide in aqueous acetone in the presence of sodium carbonate, followed by the second step to obtain the product or (tert-butyl) chlorine. or by acylation with an acylating agent of formula (V) or (VI). In the starting compound (XXII) thus obtained, X represents an oxygen atom, P) represents a (benzyloxy) carbonyl group and P ₂ represents a tert-butyldimethylsilyl group or an acyl group such as acetyl.

In the preparation of compounds of formula (XXII) wherein X is sulfur and n is 1, N, N'-bis [(benzyloxy) carbonyl] -L-cystine may be used. This is first converted to zinc powder in aqueous sulfuric acid to give the mercapto compound XLIII, and in the next step the P ₂ protecting group is introduced into the molecule, for example by reacting XLIII with acetic anhydride. In the starting compound of formula (XXII) so prepared, X is sulfur, n is 1, P _{2 is} an acetyl group, P 1 is hydrogen. and denotes benzyloxycarbonyl.

The starting compounds of formula (XXII) wherein X is sulfur and n is 2 can be prepared by, for example, protecting L-methionine with benzyl chloroformate or N - [(benzyl) -. oxy) carbonyl] succinimide, then the protected compound, N - [(benzyloxy) carbonyl] -L-methionine, is then esterified with an alcohol, preferably an alkanol, in the presence of an acid catalyst such as p-toluenesulfonic acid. In an aqueous solution of a suitable oxidizing agent such as N-chlorosuccinimide, the ester produces the corresponding sulfoxide of formula (XLIV) which gives an ester of formula (XLV) with an acid anhydride such as acetic anhydride. The reaction is carried out stepwise, first with an alkali metal hydroxide, then with a reducing agent such as sodium tetrahydroborate, to remove formaldehyde, and then with an acid anhydride, preferably acetic anhydride, to give the desired compound of formula (XXV). wherein X is sulfur, n is 2, P ₂ is acetyl, and P 1 is is (benzyloxy) carbonyl.

The fused bicyclic compounds of formula (III) wherein Y is oxygen and n is 1 can be prepared by first linking an amino acid of formula (XXII) protected by its amino group with an amino acid ester of formula (XLVI) a dipeptide of the formula XLVII, wherein P 1 and P ₂ is one of the protecting groups discussed above, and R ₃ is a carboxy protecting group. Here again, the coupling is preferably carried out with a condensing agent such as [(benzotriaziazol-1-yl) oxy] tris (dirlethylamino) phosphonium [hexafluorophosphate] or 1- [3- (dimethylamino) propyl]. In the presence of -3-ethylcarbodiimide.

Selective removal of the P ₂ protecting group, for example in methanol, sodium methylate, when P ₂ is an acyl group such as acetyl or benzoyl, or with an acid such as p-toluenesulfonic acid, also in methanol, when P _{2 is} trityl, tetrahydropyranyl or a silyl group is then converted to a compound of formula XLVIII in the presence of a suitable acid catalyst in the presence of an intermediate (XLVII).

Intermediates of formula (XLVIII) wherein X is sulfur and n is 2 can also be prepared from compounds of formula (XLVIII) wherein X is oxygen and n is 2. However, in this case, After selective removal of the P ₂ protecting group, the hydroxy group is converted to the mercapto group as previously discussed and subsequently acid-catalyzed cyclization is carried out.

Cleavage of the amino protecting group can be accomplished, for example, with hydrazine hydrate when P, together with the nitrogen atom, is phthalimido, or with iodine trimethylsilane, or in the presence of palladium on carbon, ammonium formate or hydrogen when P ₍ represents benzyloxy) carbonyl. to give the desired compound (XLVIII), the desired condensed ring compound (III).

Starting compounds of formula (XLVI) wherein Y is oxygen may be prepared starting from an amino acid ester of formula (XLIX) protected as a phthalimido group on its amino group. In the first step, the ester is reacted with allyl trichloroacetimidate in the presence of trifluoromethanesulfonic acid to form a compound of formula L, from which ozone, dimethylsulfide and finally the orthoacid ester of formula XXXII are added in the presence of p-toluenesulfonic acid. The compound of formula (LI) is obtained. The final step, removal of the amino protecting group from the molecule, which is obviously carried out here with hydrazine hydrate, affords the desired compound of formula XLVI, wherein Y is oxygen.

The present invention is illustrated by the following examples. In the specifications, the temperature is always given in ° C. Unless otherwise indicated at the site, a silica gel layer is used for TLC. [The structure of the amino compounds obtained is confirmed by the conversion of the compounds of the present invention into 4-acylamino derivatives, as disclosed in Hungarian Patent Application No. 216,791.]

Example 1 [4S- (4a, 7a, 10a, 10a)] - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] oxazepine-7-carboxylic acid methyl ester

a) Triethylamine salt of (S) -2-phthalimido-4-hydroxybutyric acid

L-homoserine (3.0 g, 25.2 mmol) and sodium carbonate (2.670 g, 25.2 mmol) were dissolved in water (60 mL) and N-ethoxycarbonylphthalimide (5.570 g, 25.4 mmol) was added. solution. The mixture was maintained for 2 hours

After stirring at room temperature, it was acidified with 6M hydrochloric acid and extracted with ethyl acetate. The ethyl acetate extract was washed with a saturated sodium chloride solution, dried over sodium sulfate, and then filtered, immediately dropping the filtrate into 4.0 mL of triethylamine in 40 mL of dichloromethane. The cloudy solution was evaporated and the residue was triturated with ethyl acetate and diethyl ether. Value of 5.11 g of a white, 142-144 ° C melting point of the resulting solid gave the title compound as a tlc _Rf 0.36 (5% acetic acid in ethyl acetate).

[α] _D = -6.2 ° (c = 0.8, chloroform).

b) Triethylamine salt of (S) -2-phthalimido-4- (triphenylmethoxy) butyric acid

To a homogeneous solution of 1.890 g (5.4 mmol) of the compound prepared in a) above in 20 ml of chloroform is added 80 µl of triethylamine followed by 1.590 g (5.70 mmol) of trityl chloride in solid form.

The reaction mixture was stirred at room temperature for 2.5 hours, then partitioned between ethyl acetate and 150 mL of 0.1 M hydrochloric acid. The organic layer was washed with water and brine, dried over sodium sulfate and filtered by passing the filtrate into 1.0 mL of triethylamine in 30 mL of dichloromethane. The solution is evaporated and the residual oil is redissolved in a little methylene chloride and ethyl acetate, then diethyl ether is added until it becomes cloudy. The solution was then inoculated and allowed to stand overnight at room temperature. The next day, the precipitate was collected by filtration, washed with ethyl acetate and diethyl ether and finally dried in vacuo. This gave 2.538 g of the title compound as white crystals. M.p. 165-170 ° C dec., TLC Rf 0.23 (10% methanol in chloroform).

[ _α ] _D = + 7.0 ° (c = 1.2, chloroform).

c) (S) -2-Phthalimido-6-hydroxyhexanoic acid

(+) -? -Hydroxy-L-norleucine (1.030 g, 7.0 mmol, prepared by Bodanszky et al., J. Med. Chem. 21, 1030-1035 (1978)) and 745 mg (7.0 mmol). sodium carbonate was dissolved in 12 mL of water and N-ethoxycarbonylphthalimide (1.495 g, 7.0 mmol) was added. The mixture was stirred at room temperature for 2 hours, then filtered, the filtrate cooled to 0 ° C and acidified with 6M hydrochloric acid. The white precipitate was collected by filtration and dried in vacuo at 80 ° C for 1 hour to give 1.297 g of the title compound. 162-163 ° C. [α] _D = -35.7 ° (c = 1.3, methanol).

d) (S) -2-Phthalimido-6,6-dimethoxy-hexanoic acid methyl ester

3.752 g (13.5 mmol) of the compound obtained in (c) above, 2.178 g (6.7 mmol) of cesium carbonate and 44 ml of N, N-dimethylformamide are weighed, and 3.0 g of this suspension are added. Methyl iodide (6.84 g, 48.2 mmol) was added. After stirring for 2 hours at room temperature, the mixture was diluted with ethyl acetate and washed successively with water containing a small amount of sodium hydrogen sulfite, clean water, half-saturated sodium bicarbonate solution and saturated sodium chloride solution. dried over sulfate, filtered and evaporated. The remaining 3,825 g of a colorless oil of the intermediate ester, homogeneous, _Rf value on which thin layer chromatography: 0: 37 (acetone: hexane = 1: 1). To a solution of oxalyl dichloride (1.37 mL, 2.00 g, 15.7 mmol) in dichloromethane (58 mL) cooled to -78 ° C was added dropwise anhydrous (2.24 g, 2.47 g, 31.6 mmol) dimethylsulfoxide and 2 mL of dichloromethane. 10 minutes later, 3.825 g (13.1 mmol) of the hydroxy ester obtained as described above are added in 10 ml of dichloromethane, and after another 15 minutes, 8.0 ml of triethylamine are added. After stirring for 5 minutes at -78 ° C, the reaction mixture was allowed to warm to room temperature, diluted with ethyl acetate / diethyl ether and washed successively with 1M hydrochloric acid, water and brine. The organic layer was dried over sodium sulfate, filtered and evaporated. The residual oil is the expected aldehyde crude product which, however, is homogeneous by TLC, Rf 0.48 (acetone: hexane = 1: 1).

The above aldehyde was dissolved in a mixture of methanol (17 ml) and dichloromethane (17 ml), followed by addition of trimethyl orthoformate (1.7 ml) followed by p-toluenesulfonic acid (180 mg, 1 molar equivalent water). The mixture was stirred at room temperature for 1.5 hours. then partitioned between ethyl acetate and half-saturated sodium bicarbonate solution. The organic layer was washed with water and saturated sodium chloride solution, dried over sodium sulfate, filtered and evaporated. The residue was purified by flash chromatography on silica gel (Merck) eluting with ethyl acetate: hexane (1: 1) and the pure fractions were recrystallized from ethyl acetate / hexane. In this way, two portions in the form of 69 to 70 ° C for melting, white needles, 3.452 g and once again 215 mg of analytically pure to give the title product having the thin layer chromatography _Rf value: 0.35 (ethyl acetate : hexane = 1: 1). [α] _D = -27.4 ° (c = 1.5, chloroform).

e) [S- (R *, R *)] - 2 - ([2-Phthalimido-4- (triphenylmethoxy) butyryl] amino) -6,6-dimethoxy-hexanoic acid methyl ester

2.540 g (7.57 mmol) of the product obtained in (d) above are suspended in 18 ml of methanol and then 378 µl (390 mg; 7.80 mmol) of hydrazine monohydrate are added. The mixture becomes homogeneous within 10 minutes. The reaction mixture is allowed to stir at room temperature for 3 days, the precipitate is filtered off, the filtrate is evaporated and the residue is suspended in dichloromethane. The resulting slurry was filtered again and evaporated to give the deprotected amine as a colorless oil.

Meanwhile, 4.622 g (7.80 mmol) of the triethylamine salt prepared in (b) are dissolved in 50 ml of methanol.

EN 217 960 Β in dichloromethane and then at 0 ° C 3.519 g (7.95 mmol) [(benzotriaziazol-1-yl) oxy] tris (dimethylamino) phosphonium hexafluorophosphate] After stirring for 35 minutes, a solution of the above amine in 15 ml of dichloromethane is added, stirring is continued for 10 minutes at 0 [deg.] C., then for 2 hours at room temperature, and then the reaction mixture is stirred between diethyl ether and water. The organic layer was washed with semi-saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate, filtered and evaporated to give a residue which was purified by flash chromatography on silica gel (Merck): ethyl acetate / hexane (6). Elution with 4 column column yields 3.580 g of a white foam which is the title product, TLC Rf 0.32 (ethyl acetate: hexane = 6: 4) [α] _D = + 26 , 2 ° (c = 0.6, chloroform).

f) [S- (R *, R *)] - 2 - [(2-Phthalimido-4-hydroxybutyryl) amino] -6,6-dimethoxyhexanoic acid methyl ester

5.420 g (8.0 mmol) of the compound prepared in (e) above and 60 ml of methanol are added and the solution is stirred at room temperature for 1.5 hours with 520 mg of p-toluenesulfonic acid water (1/1). The mixture was partitioned between ethyl acetate and dilute sodium bicarbonate solution, the immiscible phases were separated and the aqueous portion was extracted once more with ethyl acetate. The combined organic solvent extracts were washed with brine, dried over sodium sulfate, filtered and evaporated. The residue was purified by flash chromatography on silica gel (Merck) eluting the column with ethyl acetate / hexane (8: 2) followed by 5% methanol / ethyl acetate. This gave 2.860 g of the title compound as a colorless oil. TLC Rf = 0.26 (ethyl acetate: hexane = 7: 3).

[α] _D = + 18.7 ° (c = 1.3, chloroform).

g) [4S- (4a, 7a, 10a)] - 4-Phthalimido-5-oxooctahydro-7H-pyrido [2,1-b] [1,3] bxazepine-7-carboxylic acid methyl ester

Dissolve 2.10 g (4.95 mmol) of the compound prepared as in f) above in 100 mL of dichloromethane and add 240 mg of Amberlyst® 15 ion exchanger washed successively with 6 M hydrochloric acid, water, tetrahydrofuran and finally dichloromethane. resin. After stirring for 2.5 hours at room temperature, the mixture was filtered, the solution was evaporated and the residue was purified by flash chromatography on silica gel (Merck); eluting the column with ethyl acetate / hexane (6: 4) followed by pure ethyl acetate. The resulting white foam (1.40 g) is the title compound.

h) (S) -2- (Acetylthio) -3-phenylpropionic acid

D-Phenylalanine (30.0 g, 181 mmol) and potassium bromide (73.5 g) were dissolved in 2.5 M sulfuric acid (365 mL), and 10.3 g (280 mmol) was added over about 1 hour while maintaining the temperature of the solution at 0 ° C. ) sodium nitrite. The reaction mixture was stirred at 0 ° C for an additional hour, then at room temperature for a further 1 hour and then extracted with diethyl ether. The ether layer was washed once with water, dried over sodium sulfate and evaporated. The residual oil was distilled in vacuo. (R) -2-Bromo-3-phenylpropionic acid, distilled at 141 DEG C. (25.7 g) at 73 Pa (0.55 mm Hg).

[α] _D = + 14.5 ° (c = 2.4, chloroform).

Thioacetic acid (7 mL, 97.9 mmol), potassium hydroxide (5.48 g, 97.9 mmol) and acetonitrile (180.5 mL) were added and the mixture was stirred under argon at room temperature for 1.75 hours. After cooling in an ice bath, a solution of (R) -2-bromo-3-phenylpropionic acid (20.4 g, 89 mmol) in acetonitrile (20 mL) was added with cooling over a period of about 10 minutes. Stirring is continued at room temperature under argon for 5 hours, then the mixture is filtered and the acetonitrile is evaporated in vacuo. The oily residue was redissolved in ethyl acetate, washed with 10% potassium bisulfate solution and water and then concentrated in vacuo. The crude product (19.6 g) was purified via its dicyclohexylamine salt to remove the salt from diisopropyl ether solution. An analytical sample of the dicyclohexylamine salt of (S) -2-acetylthio-3-phenylpropionic acid was recrystallized from ethyl acetate, m.p. 146-147 ° C. [α] _D = -39.6 ° (c = 1.39, chloroform).

The free acid from the dicyclohexylamine salt is recovered by shaking with a mixture of 5% potassium bisulfate solution and ethyl acetate. The product liberated from the salt is (S) -2- (acetylthio) -3-phenylpropionic acid. [α] _D = -70.1 ° (c = 1.91, chloroform).

i) [4S- [4a (R *), 7x, 10a-4]] - 4 - {[2- (Acetylthio) -3-phenylpropionyl] amino} -5-oxo-octahydro- 7H-Pyrido [2,1b] [1,3] oxazepine-7-carboxylic acid methyl ester

620 mg (1.66 mmol) of the compound obtained in (g) above, 10 ml of methanol and 85 μΐ (88 mg; 1.75 mmol) of hydrazine monohydrate are weighed. After stirring for 44 hours at room temperature, the mixture was filtered and the filtrate washed with methanol. The filtrate was evaporated and the residue was triturated with dichloromethane and then filtered again. The solvent was evaporated to give 400 mg of [4S- (4a, 7a, 10ap)] - 4-amino-5-oxo-octahydro-7H-pyrido [2,1b] [1,3] oxazepine-7-carboxylic acid methyl acetate as a cloudy oil. ester is obtained.

(S) -2-Acetylthio-3-phenylpropionic acid (410 mg, 1.83 mmol) and triethylamine (250 μΐ, 182 mg, 1.80 mmol) were dissolved in dichloromethane (10 mL), then 0 °. At C, a solution of the above amine in 8 mL of dichloromethane was added, followed by 808 mg (1.83 mmol) of [(benzotriazol-1-yl) oxy] tris (dimethylamino) phosphonium [hexafluoro -phosphate] is added. The resulting clear, nearly colorless solution ^J 0 C then at room temperature for 2 hours 40 minutes, then ethyl acetate, between diethyl ether and water and partitioned. The organic phase was washed successively with half-saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate and evaporated. The residue was purified by flash chromatography on silica gel (Merck)

HU 217 960 Β

Eluting the column with 60-70% ethyl acetate / hexane. The resulting white foam (602 mg) is the title compound in pure form. The product TLC _Rf value: 0.27 (ethyl acetate: hexane = 6: 4).

(The product was thus used as {4S- [4a (R *), 7a, 10aP]} - 4 - [(3-phenyl-2-mercaptopropionyl) amino] -5-oxooctahydro-7H-pyrido [2 1b] [1,3] oxazepine-7-carboxylic acid, the structure of which has been verified.

Example 2 [3R- (3a, 6a, 9a, 9a)] - 3-Amino-4-oxo-hexahydro-2H, 6H-pyrido [2,1-b] [1,3] thiazine-6-carboxylic acid methyl ester

a) N - [(Benzyloxy) carbonyl] -L-cysteine

N, N'-bis [(benzyloxy) carbonyl] -L-cystine (4.658 g, 9.16 mmol) was dissolved in methanol (35 mL) followed by 2 M sulfuric acid (23 mL) followed by zinc powder (2.442 g, 37.3 mmol). is added to the solution. The reaction mixture was heated to 70 ° C and held at this temperature for 1.5 hours, then filtered hot while the filtrate was concentrated on a rotary evaporator. The resulting solution was extracted with diethyl ether, and the ether extract was washed with water and brine, dried over sodium sulfate, filtered and the solvent was evaporated. The evaporation residue is an oil which is dissolved in carbon tetrachloride. The solution was cooled to 0 ° C and inoculated, which resulted in slow precipitation. The precipitate was filtered off and washed with cold carbon tetrachloride. It is a first generation product weighing 2,648 g. The mother liquor was evaporated and the residue was purified by flash chromatography on silica gel (Merck), eluting the column first with ethyl acetate and then with 4% acetic acid / ethyl acetate. The material thus purified was recrystallized to give an additional 246 mg of the title compound. This second generation product was combined with the first generation product to give a total yield of 2.894 g. The pure product by thin layer chromatography _Rf value: 0.58 (5% acetic acid in ethyl acetate).

b) S-Acetyl-N - [(benzyloxy) carbonyl] -L-cysteine

Purge with argon gas to dissolve dissolved oxygen in 30 mL of water, potassium bicarbonate (2.140 g, 21.4 mmol), then 2.70 g (10.6 mmol) of the compound prepared in (a) above, then add to the homogeneous solution. Acetic anhydride (8.0 mL, 8.66 g, 84.8 mmol) was added. After 10 minutes at room temperature, the mixture was acidified with 10% hydrochloric acid and extracted with diethyl ether. The ether extract was washed twice with water and a saturated sodium chloride solution twice, dried over sodium sulfate, filtered and evaporated. The residual oil was azeotroped three times with toluene and twice with ethyl acetate / hexane, whereupon the oil crystallized. The crystals were triturated with ethyl acetate / hexane and then collected on a filter. 2.19 g of pure product is thus obtained the title compound as a tlc _Rf 0.56 (5% acetic acid in ethyl acetate).

c) Methyl (S) -2 - [{S-Acetyl-N - [(benzyloxy) carbonyl] -L-cysteinyl} -amino] -6,6-dimethoxy-hexanoic acid

1.158 g (3.45 mmol) of (S) -2-phthalimido-6,6-dimethoxy-hexanoic acid methyl ester prepared as described in Example 1 (d) are suspended in 12 ml of methanol. To the suspension was added 176 µl (182 mg, 3.63 mmol) of hydrazine monohydrate, which resulted in a homogeneous mixture within 10 minutes. After 67 hours at room temperature, the precipitate was filtered off and the filtrate was evaporated. The residue was suspended in dichloromethane, filtered again, and the solvent was evaporated to give the deprotected intermediate as a colorless oil.

Meanwhile, 1.185 g (3.98 mmol) of the product prepared as in b) above are weighed; and dichloromethane (14 ml), then triethylamine (555 µl, 403 mg, 3.98 mmol) was added to the resulting partial suspension. The resulting homogeneous solution was cooled to 0 ° C and a solution of the above amine in 7 mL of methylene chloride was added. Subsequently, 1.762 g (3.98 mmol) of [(benzotriazol-1-yl) oxy] tris (dimethylamino) phosphonium [hexafluorophosphate] were added, followed by 0 ° C for 2.5 hours. After stirring at room temperature for 45 minutes, the reaction mixture was evaporated and the residue was partitioned between ethyl acetate and water. The organic layer was washed with semi-saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate, filtered, and then the solvent was evaporated. The resulting crude product was purified by flash chromatography on silica gel (Merck) eluting the column with 65:35 ethyl acetate: hexane. 1.15 g of a white foam are thus obtained, which is the pure product of the title compound, TLC Rf 0.42 (ethyl acetate: hexane = 75:25).

d) [3R- (3a, 6a, 9a)] - 3 - {[(Benzyloxy) carbonyl] amino} -4-oxo-hexahydro-2H, 6H-pyrido [2,1-b] [1 , 3] thiazine-6-carboxylic acid methyl ester

To a solution of 1.040 g (2.15 mmol) of the compound prepared in (c) above in a solution of 12 ml of methanol in argon gas was degassed with oxygen and cooled to 0 ° C with 490 μΐ (463 mg, 2.14 mmol) of 25% w / w sodium methylate. methanol solution. After 20 minutes, the reaction was quenched with saturated ammonium chloride, diluted with water and extracted with ethyl acetate. The ethyl acetate extract was washed with water and saturated sodium chloride solution, dried over sodium sulfate, filtered and evaporated. The residue was redissolved in CH 2 Cl 2 (200 mL), and the solution was stirred for 3 hours after addition of 820 mg of Amberlyst® 15 ion exchange resin washed successively with 6 M hydrochloric acid, water, tetrahydrofuran and finally CH 2 Cl 2. After the reaction time, the solution is filtered, the solvent is evaporated and the crude product is purified by flash chromatography on silica gel (Merck) with ethyl acetate and hexane.

HU 217 960 Β

Eluting with a 65:35 mixture. In this way 757 mg of a colorless oil of the title compound, the product TLC _Rf value: 0.58 (ethyl acetate: hexane = 75: 25).

e) [3R- (3a, 6a, 9a)] - 3-Amino-4-oxo-hexahydro-2H, 6H-pyrido [2,1-b] [1,3] thiazine-6-carboxylic acid methyl ester

Dissolve 752 mg (1.99 mmol) of the compound obtained in (d) above in 15 ml of anhydrous dichloromethane and add i20-trimethylsilane (620 µL, 872 mg, 4.36 mmol) at room temperature. After stirring for 3 hours, the mixture was diluted with water and extracted with a little 10% hydrochloric acid and extracted with diethyl ether. The immiscible layers are separated and the organic phase is back-extracted with water. The combined aqueous layer was basified to pH 13 with 10% sodium hydroxide solution and extracted twice with methylene chloride twice. The methylene chloride solutions were combined, dried over sodium sulfate, filtered and evaporated. The value of the resulting 290 mg of colorless oil of the title crude product as a TLC _Rf: 0.38 (10% methanol in methylene chloride).

(From the product {3R- [3a (S *), 6a, 9aP]} - 3 - ([2- (acetylthio) -3-phenylpropionyl] amino} -4-oxo-hexahydro-2H, 6H-pyrido [2,1-b] [1,3] thiazine-6-carboxylic acid methyl ester followed by {3R- [3a (S *), 6a, 9ap]} - 3 - [(3-phenyl-2-mercaptopropionyl) ) -amino] -4-oxo-hexahydro-2H, 6H-pyrido [2,1b] [1,3] thiazine-6-carboxylic acid, the structure of which has been verified.

Example 3 {4S- [4a, 7a, 10a-$]} - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

a) [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2-phthalimido-butyryl] -amino} -6,6-dimethoxy-hexanoic acid, methyl ester

Triphenylphosphine (1.143 g, 4.36 mmol) was dissolved in tetrahydrofuran (20 mL) and diisopropyl azodicarboxylate (860 µL, 883 mg, 4.37 mmol) was added to the solution cooled to 0 ° C over 5 minutes. After 30 minutes, [S (R *, R *)] - 2 - [(2-phthalimido-4-) (928 mg, 2.19 mmol) prepared as in Example 1 (f) was added. hydroxybutyryl) amino] -6,6-dimethoxy-hexanoic acid methyl ester in 8 ml of tetrahydrofuran, then 312 [mu] l (332 mg, 4.36 mmol) of thioacetic acid are added and the mixture is stirred at 0 [deg.] C. for 1.25 hours. let it mix. After the reaction time, the mixture was partitioned between ethyl acetate and half-saturated sodium bicarbonate solution. The separated ethyl acetate layer was washed with a saturated sodium chloride solution, dried over magnesium sulfate and evaporated. The residue was redissolved in ethyl acetate, a little hexane was added and the precipitated triphenylphosphinoxide precipitate was filtered off and the filtrate was purified by flash chromatography on silica gel (Merck) eluting with 65:35 ethyl acetate: hexane. The title compound was obtained as a colorless oil (894 mg). The product TLC _Rf value: 0.43 (ethyl acetate: hexane = 75: 25).

b) [4S- (4a, 7a, 10a)] - 4-Phthalimido-5-oxooctahydro-7H-pyrido (2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

To a solution of the compound (a) (814 mg, 1.65 mmol) in methanol (15 mL), purged with argon gas and cooled to 0 ° C was added a methanol solution (1.05 mL, 4.6 mmol) containing 25% by weight sodium methylate. . After 5 minutes, the reaction was quenched with saturated ammonium chloride solution, diluted with water and extracted with ethyl acetate. The ethyl acetate extract was washed with water and saturated sodium chloride solution, dried over sodium sulfate, filtered and the solvent was evaporated. The residue was redissolved in dichloromethane (180 ml) and treated with Amberlyst® 15 ion exchange resin (285 mg, successively washed with 6M hydrochloric acid, water, tetrahydrofuran, and finally dichloromethane) and stirred at room temperature for 46 hours. After the reaction time, the solution was filtered, the filtrate was evaporated and the crude residue was purified by flash chromatography on silica gel (Merck) eluting with 1: 1 ethyl acetate: hexane. This gave 314 mg of the title compound as a white foam. The foam was triturated with ethyl acetate is converted into a white powdery solid melting at 147-148 ° C, TLC _Rf value: 0.56 (ethyl acetate: hexane = 75: 25).

[α] _D = -143.2 ° (c = 0.6, chloroform).

c) {4S- [4a, 7a, 10a-$]} - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

To 280 mg (0.72 mmol) of the compound prepared as in (b) above was added methanol (8 mL) followed by hydrazine monohydrate (92 µL, 43.3 mg, 0.86 mmol). The mixture was allowed to stir at room temperature for 67 hours, then filtered and the precipitate was washed with methanol. The filtrate was evaporated, the residue was triturated with dichloromethane, filtered again and the solvent was evaporated. The yellow oil thus obtained, about 205 mg, was a crude amino compound deprotected from the phthaloyl protecting group.

(From the product {4S- [4a (R *), 7a, 10aP]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydro-7H-pyrido [2 , 1b] [1,3] Thiazepine-7-carboxylic acid methyl ester {4S- [4a (R '), 7a, 10ap]} - 4 - [(3-phenyl-2-mercaptopropionyl) - amino] -5-oxooctahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid was isolated and isolated.

Example 4 [4S- (4a, 7a, 9a ')] - 4-Amino-5-oxo-octahydro-pyrrolo [2,1-b] [1,3] oxazepine-T-carboxylic acid methyl ester

a) (S) -4 - [(Benzyloxy) carbonyl] -2-phthalimidobutyric acid

17.49 g (73.70 mmol) of glutamic acid γ-benzyl ester, 180 ml of water, 7.81 g (73.70 mmol) of sodium carbonate and 120 ml of dioxane are weighed, and 16.50 g (75 ml) of (27 mmol; 1.02 eq.) Of N- (ethoxycarbonyl) phthalimide was added and the mixture was stirred at room temperature for 4.5 hours, then acidified with 30 ml of 6M hydrochloric acid and extracted with 2 x 400 ml of ethyl acetate.

HU 217 960 Β net. The combined ethyl acetate extracts were washed with half-saturated sodium chloride solution (200 mL) and brine (200 mL), dried over sodium sulfate, filtered and evaporated. The residual oil was dried in vacuo to give the crude product (41.4 g) which was dissolved in diethyl ether (100 mL) and treated with dicyclohexylamine (14 mL). After standing overnight in a refrigerator, the next day, the diethyl ether was evaporated in a rotary evaporator and the residual oil was crystallized from ethyl acetate / hexane. The crystals were transferred to a filter, washed with hexane and dried in vacuo to afford

21.21 g of salt are obtained, this salt being the dicyclohexylamine salt of the title compound. The dicyclohexylamine salt is suspended in 200 ml of ethyl acetate, shaken first with 3 times 50 ml of 5% potassium bisulfate solution, and the separated organic phase is washed with 50 ml of saturated sodium chloride solution and dried over magnesium sulphate. , filtered and the solvent is evaporated. 13.5 g of a white foam are obtained in the form of the title compound. The product TLC _Rf value: 0.30 (3% acetic acid in ethyl acetate and heptane 9: 1 mixture).

b) Methyl (S) -4 - [(benzyloxy) carbonyl] -2-phthalimidobutyric acid

Dissolve in 100 ml of Ν, Ν-dimethylformamide

13.22 g (36.0 mmol) of the compound prepared in a) above, followed by 5.86 g (18.0 mmol) of cesium carbonate, followed by 8.1 ml (129.6 mmol; 3.6 eq.) ) methyl iodide was added to the solution. The yellowish mixture was stirred for 2.5 hours, then partitioned between 300 mL of ethyl acetate and 250 mL of water. The ethyl acetate solution was washed with 200 ml of 5% sodium bicarbonate solution and saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. The residual yellow oil (13.68 g) was purified by column chromatography over silica gel (5 x 20 cm) eluting with 30% ethyl acetate / hexane. The appropriate fractions were combined and the solvent was evaporated to give 10.0 g of the title compound. The product TLC _Rf value: 0.45 (ethyl acetate: hexane = 1: 1).

c) Methyl (S) -2-phthalimido-4-carboxylbutyric acid

The ester obtained in (b) above (10.0 g, 26.22 mmol) was dissolved in ethyl acetate (115 mL), palladium hydroxide (20%, charcoal precipitated) (1.90 g) was added, and the suspension was stirred under hydrogen atmosphere. , Stir for 5 hours. After the reaction time, the catalyst was filtered off, rinsed thoroughly with ethyl acetate, and the filtrate was concentrated in vacuo. The title compound was obtained as a white solid (7.29 g), m.p. 137-138 ° C. The product TLC _Rf value: 0.43 (10% methanol in methylene chloride).

d) Methyl (S) -4 - [(ethylthio) carbonyl] -2-phthalimidobutyric acid

7.27 g (24.95 mmol) of the compound obtained in (c) above are dissolved in 125 ml of dichloromethane, the solution is cooled to 0 ° C, and 4.81 ml (64 , 92 mmol; 2.6 eq.) Ethanethiol, 609 mg (4.99 mmol; 0.2 eq.) Of 4- (dimethylamino) pyridine and 5.27 g (27.47 mmol; 1.1 eq.) Of 1- [ 3- (dimethylamino) propyl] -3-ethylcarbodiimide hydrochloride. The reaction mixture was stirred at 0 ° C for 2 hours; After stirring for a further 1 hour at room temperature, the mixture was evaporated, the residue was taken up in 400 ml of ethyl acetate and 200 ml of 5% potassium bisulfate solution, 200 ml of saturated sodium bicarbonate solution and finally 200 ml. washed with saturated sodium chloride solution. The organic layer was dried over sodium sulfate, filtered and the solvent was evaporated in vacuo. The value thus obtained 8.30 g of an oil of the title compound, the product TLC _Rf: 0.47 (ethyl acetate: hexane = l: l).

e) Methyl (S) -4-Formyl-2-phthalimidobutyric acid

8.30 g (24.75 mmol) of the compound prepared in (d) above and 1.24 g of 10% palladium on charcoal are suspended in 150 ml of acetonitrile and then 7.91 ml (49.5 mmol) of argon are added dropwise under argon. 2.0 eq.) Of triethylsilane. The reaction mixture was stirred for 45 minutes at room temperature, then filtered, concentrated in vacuo and dried. The crude product was purified by chromatography on a 5 x 25 cm silica gel column eluting with 4000 ml of 25% ethyl acetate / 75% hexane, then 2000 ml of 35% ethyl acetate / 65% hexane. The appropriate fractions gave 5.60 g of the title compound. The product TLC _Rf: 0.32 (ethyl acetate: hexane = 1: 1).

j) (S) -4-Phthalimido-5,5-dimethoxy-valeric acid methyl ester

5.60 g (20.34 mmol) of the compound prepared as in e) above are dissolved in 60 ml of methanol and 40 ml of dichloromethane, followed by 3.8 ml (34.59 mmol; 1.7 equivalents) of trimethyl orthoformate. and 280 mg of p-toluenesulfonic acid water (1/1) were added to the solution

Stir at room temperature for 1.5 hours. The reaction mixture was quenched with saturated sodium bicarbonate solution (2 mL), evaporated and partitioned between ethyl acetate (400 mL) and water (400 mL). The ethyl acetate layer was washed with 100 mL of saturated sodium bicarbonate solution and 100 mL of saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. The residual oily crude product is purified by chromatography on a 5 x 20 cm silica gel column eluting with 2000 ml of a mixture of 30% ethyl acetate and 70% hexane. The appropriate fractions were combined and concentrated in vacuo and dried to give 6.20 g of the title compound. The product TLC _Rf = 0.40 (ethyl acetate: hexane = 1: 1).

g) (S) -2-Amino-5,5-dimethoxy-valeric acid methyl ester

6.16 g (19.18 mmol) of the product obtained above under f) are dissolved in 125 ml of methanol 9

After addition of hydrazine monohydrate (0.98 mL, 20.14 mmol, 1.05 equiv). After stirring for 6 days, the resulting suspension is filtered, the filtrate is evaporated and the residue is triturated with methylene chloride. The solid precipitate was filtered off again and the solvent was evaporated in vacuo. The resulting compound (3.57 g) is a cloudy oil. The product TLC _Rf value: 0.41 (10% methanol in methylene chloride).

h) [S- (R *, R *)] - 2 - {[2-phthalimido-4- (triphenylmethoxy) butyryl] amino} -5,5-dimethoxy-valeric acid methyl ester

11.62 g (19.60 mmol; 1.05 equivalents), Example 1

(S) -2-Phthalimido-4-triphenylmethoxy-butyric acid (triethylamine) salt prepared in (b) is dissolved in 100 ml of dichloromethane and 8.67 g of the solution cooled to 0 ° C are obtained. 19.60 mmol (1.05 equiv) of (benzotriaziazol-1-yl) oxy-tris (dimethylamino) phosphonium hexafluorophosphate. After stirring for 45 minutes at 0 ° C, a solution of 3.57 g (18.67 mmol) of the above compound (g) in 50 ml of dichloromethane is added. After stirring for 10 minutes at 0 ° C and for 2 hours at room temperature, the reaction mixture was partitioned between 300 ml of ethyl acetate and 100 ml of water. The ethyl acetate layer was washed with 100 ml of semi-saturated sodium bicarbonate solution and saturated sodium carbonate solution, dried over magnesium sulfate, filtered and evaporated, and the residue was purified by chromatography on a 5 x 25 cm silica gel column. eluting the column with ethyl acetate / hexane (1: 1). In this way 8.58 g of the title: product with a TLC _Rf value: 0.20 (ethyl acetate: hexane = 1: 1).

i) [S- (R *, R *)] - 2 - [(2-Phthalimido-9-hydroxybutyryl) amino] -5,5-dimethoxy-valeric acid methyl ester

8.58 g (12.91 mmol) of the product obtained above under h) are dissolved in 100 ml of methanol and then 850 mg of p-toluenesulfonic acid water (1/1) are added and the solution is stirred at room temperature for 3.5 hours. After the reaction time, the mixture was partitioned between ethyl acetate (200 mL) and saturated sodium bicarbonate solution (10%, 100%). The two phases are separated and the aqueous phase is extracted once more with 100 ml of ethyl acetate. The combined ethyl acetate extracts were washed with saturated sodium chloride solution, dried over magnesium sulfate, filtered, and the residue was purified by chromatography on a 5 x 20 cm silica gel column, first with 1000 ml of 8: 2 ethyl acetate: hexane and then with 2,000 mL of 1% methanol in ethyl acetate. The appropriate fractions were combined, the solvent evaporated in vacuo and the residue dried. 4.33 g of the title compound are obtained with TLC Rf = 0.21 (ethyl acetate: hexane = 8: 2).

j) [4S- (4a, 7a, 9a ')] - 4-Phthalimido-5-oxo-octahydro-pyrrolo [2,1-b] [1,3] oxazepine-7-carboxylic acid methyl ester

To a solution of 1.89 g (4.48 mmol) of the compound obtained in (i) above in 90 ml of dichloromethane is added 400 mg of Amberlyst® 15 ion exchange resin, successively with 6 M hydrochloric acid, water, tetrahydrofuran and finally methylene chloride. washed thoroughly with dichloride. After stirring for 3 hours at room temperature, the filtrate was evaporated and the residue was purified by flash chromatography on a silica gel column (5 x 15 cm) eluting with ethyl acetate / hexane (6: 4). The resulting white foam (1.51 g) is the title compound. The product TLC _Rf: 0.32 (ethyl acetate: hexane = 8: 2).

k) [4S- (4a, 7α, 9αβ)] - 4-Amino-5-oxo-octahydro-pyrrolo [2,1b] [1,3] oxazepine-7-carboxylic acid methyl ester

764 mg (2.13 mmol) of the compound prepared as in (j) above are added, 15 ml of methanol are added, and 109 μΐ (2.24 mmol; 1.05 equivalents) of hydrazine monohydrate are added and the mixture is stirred for 4 days at room temperature. stirred. After the reaction time, the precipitate was filtered off, washed with methanol, the filtrate was evaporated and the residue was triturated with methylene chloride. The resulting suspension is filtered again, then the solvent is evaporated and the crude residue is purified by chromatography on a 2 x 15 cm silica gel column, first with 3000 mL of 3% methanol and 10% methanol in dichloromethane and 10% methanol in dichloromethane. the column. The appropriate fractions were combined and evaporated to give 451 mg of the title compound. The product is an oil having a thin layer chromatography _Rf value: 0.18 (10 methanol in dichloromethane).

(From the product {4S- [4a (R *), 7a, 9a3]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydropyrrolo [2,1-b ] [1,3] oxazepine-7-carboxylic acid methyl ester {4S- [4a (R *), 7a, 9ap]} - 4 - [(3-phenyl-2-mercaptopropionyl) amino] - 5-oxo-octahydropyrrolo [2,1-b] [1,3] oxazepine-7-carboxylic acid was prepared and its structure was confirmed.]

Example 5 [4S- (4a, 7a, 9aβ)] - 4-Amino-5-oxo-octahydro-pyrrolo [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

a) [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2-phthalimido-butyrylamino] -5,5-dimethoxy-valeric acid methyl ester

To a solution of triphenylphosphine (1.26 g, 4.79 mmol, 1.5 equiv.) In dry tetrahydrofuran (15 mL) cooled to 0 ° C was added diisopropyl azodicarboxylate (943 μΐ, 4.79 mmol). The resulting white suspension was stirred for 30 minutes and then added with [S- (R *, R *)] - 2 - [(2-phthalimido-4) (1.35 g, 3.20 mmol) prepared as in Example 4 (i). -hydroxybutyryl) amino] -5,5-dimethoxy-valeric acid methyl ester in 15 ml of anhydrous tetrahydrofuran. After addition of 343 μ 4 (4.79 mmol) of thioacetic acid, the mixture was stirred at 0 ° C for 1.5 hours and then partitioned between 150 ml of ethyl acetate and 100 ml of semi-saturated sodium bicarbonate solution. The ethyl acetate layer was washed with a saturated sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated to remove the residue.

EN 217 960 Β adsorbed on Celite and kept under vacuum until dry. The crude product was purified by chromatography on a 2.5 χ 15 cm silica gel column, eluting with 1000 ml of ethyl acetate: hexane 1: 1 and then 1000 ml of ethyl acetate: hexane 6: 4. The appropriate fractions were combined and the solvent was evaporated in vacuo. 1.35 g of an oil are thus obtained which is the title compound. TLC Rf = 0.42 (ethyl acetate: hexane = 8: 2).

b) [S- (R *, R *)] - 2 - [(Phthalimido-4-mercaptobutyryl) amino] -5,5-dimethoxy-valeric acid methyl ester

1.33 g (2.76 mmol) of the compound prepared as in a) above are dissolved in methanol and the solution is degassed by bubbling argon gas. Subsequently, 1.52 ml (6.63 mmol; 2.4 equivalents) of 25% (w / w) sodium methoxide in methanol are added at 0 [deg.] C., and the reaction is quenched 3 minutes later with 3 ml of saturated ammonium chloride solution. . The mixture was diluted with water, extracted with ethyl acetate (100 mL), and the ethyl acetate extract was washed with water (50 mL) and brine (50 mL), dried over sodium sulfate, filtered and evaporated. The residue was purified by chromatography on a 5 x 15 cm silica gel column, eluting with 3000 mL of 1: 1 ethyl acetate: hexane and then 2000 mL of 8: 2 ethyl acetate: hexane. The appropriate fractions were combined and the solvent was evaporated to give 853 mg of the title compound. The product is an oil having a thin layer chromatography _Rf value: 0.43 (ethyl acetate: hexane = 8: 2).

c) [4S- (4a, 7a, 9a, 8a)] - 4-Phthalimido-5-oxo-octahydro-pyrrolo [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester is dissolved in dichloromethane (847 mg). (93 mmol) of the compound prepared as in (b) above, to the solution was added 700 mg of Amberlyst (R) 15 ion exchange resin, which was thoroughly washed successively with 6 M hydrochloric acid, water, tetrahydrofuran and finally dichloromethane. After stirring at room temperature for 17 hours, the reaction mixture was filtered, the filtrate was evaporated and the residue was purified by flash chromatography on a 2.5 x 15 cm silica gel column eluting with ethyl acetate: hexane (1: 1). column. The title compound was obtained as a white foam (691 mg). TLC Rf 0.48 (ethyl acetate: hexane = 8: 2).

d) [4S- (4a, 7a, 9a ')] - 4-Amino-5-oxo-octahydropyrrolo [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

To the product obtained in (c) above (899 mg, 2.40 mmol) was added methanol (17 mL) and hydrazine monohydrate (122 µl, 2.52 mmol, 1.05 equivalent). After stirring at room temperature for 3 days, the precipitate was filtered off and washed with methanol. The filtrate was evaporated and the residue was triturated with dichloromethane and then filtered again. Evaporation of the solvent and drying of the product in vacuo gave 572 mg of the title compound as an opaque oil with TLC Rf 0.13 (10% methanol in dichloromethane).

(From the product {4S- (4a, R *), 7a, 9aP]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydropyrrolo [2,1b] [ 1,3] thiazepine-7-carboxylic acid methyl ester and {4S [4a (R *), 7a, 9aP]} - 4 - [(3-phenyl-2-mercaptopropionyl) amino] -5-oxo octahydropyrrolo [2,1b] [1,3] thiazepine-7-carboxylic acid was prepared and verified.

Example 6 [4S- (4a, 7a, 9αβ)] - 4-Amino-5-oxo-octahydro-pyrrolo [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

In addition to that described in Example 5, the title compound can be prepared as follows:

a) N - [(Benzyloxy) carbonyl] -L-homoserine

Dissolve 10.24 g (85.98 mmol) of L-homoserine and 7.95 g (94.58 mmol; 1.1 equivalents) of sodium bicarbonate in 100 mL of water and 100 mL of acetone, followed by 23.57 g ( N-{[(benzyloxy) carbonyl] oxy} succinimide (94.58 mmol) was added to the solution. After stirring overnight at room temperature, the next day, the acetone was evaporated in vacuo on a rotary evaporator and the resulting aqueous solution was washed with methylene chloride (2 x 75 mL). The aqueous phase was then acidified to pH 2 with 6M hydrochloric acid and then extracted with ethyl acetate (2 x 250 mL). The combined ethyl acetate extracts were washed with water (2 x 100 mL) and brine, dried over sodium sulfate, filtered and evaporated. The value thus obtained 19.54 g of white solid of the title compound, TLC _Rf (1.74 (AcOEt: n-butanol: acetic acid: water = 2: 1: 1: 1).

b) N - [(Benzyloxy) carbonyl] -O- (triphenylmethyl) -Lhomoserine

To a suspension of 19.51 g (77.04 mmol) of the compound prepared in (a) above in 250 ml of chloroform is added 12.35 ml (88.59 mmol; 1.15 equivalents) of triethylamine and the resulting homogeneous solution 24.70 g (88.59 mmol) of triphenylmethyl chloride are added. After stirring for 3 hours, the reaction mixture was concentrated in vacuo on a rotary evaporator and the residue was partitioned between ethyl acetate (400 mL) and 5% potassium bisulfate (200 mL). The ethyl acetate layer was washed with 200 mL of 5% potassium bisulfate, 2 x 200 mL of water and 200 mL of saturated sodium chloride solution, dried over sodium sulfate, filtered and evaporated. The crude product (45.4 g) was purified by column chromatography over silica gel (10 x 30 cm), eluting first with 2000 mL of 6: 4 ethyl acetate: hexane and then 1: 8 acetic acid: ethyl acetate: hexane. the column. This gives 8.76 g of the pure title compound.

c) Methyl (S) -2-amino-5,5-dimethoxy-valeric acid

3.35 g (10.43 mmol) of (S) -2-phthalimido-5,5-dimethoxy-valeric acid methyl ester prepared as in Example 4 (f) were treated with 70 ml of methanol and 531 μΐ.

Hydrazine monohydrate (10.95 mmol, 1.05 eq.) Was added and the resulting solution was stirred at room temperature for 6 days. After the reaction time, the mixture was filtered, the solid was washed with methanol and the filtrate was evaporated. The residue was triturated with dichloromethane, filtered again, the solvent evaporated and the residue dried in vacuo. The value thus obtained 1.89 g of a cloudy oil gave the title compound as a tlc _Rf 0.39 (10% methanol in dichloromethane).

d) [S- (R *, R *)] - 2 - {[2 - {(Benzyloxy) carbonyl] amino} -4- (triphenylmethoxy) butyryl] amino} -5.5 dimethyl valeric acid methyl ester

5.28 g (10.65 mmol, 1.1 equivalents) of the compound prepared as in b) above are dissolved in 50 ml of anhydrous dichloromethane and 1.48 ml (10.65 mmol) are added at 0 ° C. triethylarlin was added and a solution of 1.85 g (9.68 mmol) of the above compound (c) in 30 ml of dichloromethane was added. Subsequently, 4.71 g (10.65 mmol; 1.1 equivalents) of [(benzotriazol-1-yl) oxy] tris (dimethylamino) phosphonium [hexafluorophosphate] are added, followed by The mixture was allowed to stir at 0 ° C for 1 hour and then at room temperature for a further 2 hours. After the reaction time, the mixture was partitioned between ethyl acetate (300 mL) and water (150 mL), and the separated ethyl acetate layer was washed with half saturated sodium bicarbonate solution (200 mL) and brine (2 x 200 mL), magnesium sulfate. dried, filtered and finally concentrated. The residue was adsorbed onto Celite and loaded onto a 7 x 20 cm silica gel column, eluting first with 3000 mL of 90% ethyl acetate: hexane followed by 2000 mL of 50% ethyl acetate: hexane. In this manner 4.84 g of title named product having a thin layer chromatography _Rf value: 0.22 (ethyl acetate: hexane = 1: 1).

e) [S- (R *, R *)] - 2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4-hydroxybutyryl] amino} -5,5-dimethoxy-valeric acid methyl -ester

Dissolve 4.80 g (7.18 mmol) of the compound prepared as in (d) above in 70 ml of methanol, add 300 mg of p-toluenesulfonic acid water (1/1) and stir at room temperature for 2 hours, then add 400 ml of ethyl acetate. acetate and 200 ml of 25% saturated sodium bicarbonate solution. The immiscible phases were separated, the aqueous portion was extracted with 100 mL of ethyl acetate, and the combined ethyl acetate extracts were washed with brine, dried over magnesium sulfate, filtered and evaporated. The residue was purified by chromatography on a 5 x 20 cm silica gel column, first with 1000 ml of ethyl acetate: hexane (7: 3), then with 1000 ml of ethyl acetate: hexane (8: 2). eluting the column with a mixture of methanol and ethyl acetate. The appropriate fractions were combined, the solvent was evaporated and the residue was dried in vacuo. This residue, weighing 2.92 g, was the title compound. The product TLC _Rf value: 0.09 (ethyl acetate: hexane = 8: 2).

fi [S- (R *, R * j] -2 - {[4- (Acetylthio) -2 - {[(benzyloxy) carbonyl] amino} butyryl] amino} -5,5- dimethoxypentanoic acid, methyl ester

To a solution of triphenylphosphine (3.06 g, 11.65 mmol) in dry tetrahydrofuran (40 mL) cooled to 0 ° C

Diisopropyl azodicarboxylate (2.29 mL, 11.65 mmol) was added. The resulting white slurry was stirred for 30 minutes, then dissolved in anhydrous tetrahydrofuran (2.92 g, 6.85 mmol) prepared in (e) above and 833 μ33 (11.65 mmol) of thioacetic acid. Stirring was continued at 0 ° C for an additional 2 hours, then the mixture was partitioned between 300 mL of ethyl acetate and 200 mL of semi-saturated sodium bicarbonate solution. The separated ethyl acetate layer was washed with a saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. The residual crude product was adsorbed onto Celite and dried in vacuo on a 5 x 20 cm silica gel column. The column was eluted with 3000 mL of 1: 1 ethyl acetate: hexane, then the pure fractions were combined, the solvent was evaporated, and the residue was dried in vacuo. The resulting 2.58 g of off-white product having a tlc _Rf 0.40 (ethyl acetate: hexane = 8: 2) to give the title compound.

g) [S - (R *, R *)] - 2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4-mercaptobutyryl] amino} -5,5-dimethoxyveric acid methyl ester

To a solution of 2.56 g (5.28 mmol) of the compound prepared as in (f) above in 50 mL of methanol, de-oxygenated by bubbling argon gas and cooled to 0 ° C, was added 3.62 mL (15.84 mmol; 3 equivalents). sodium methoxide in methanol by weight. After 10 minutes, the reaction was quenched with saturated ammonium chloride (40 mL), diluted with water (100 mL) and extracted with ethyl acetate (300 mL). The ethyl acetate extract was washed with water (100 mL) and brine (150 mL), dried over magnesium sulfate, filtered, evaporated and the residue was purified by chromatography on a silica gel column (5 x 20 cm). The column was eluted with 3000 mL of 1: 1 ethyl acetate: hexane, and the appropriate fractions were combined and the solvent was evaporated. The resulting oil (1.99 g) is obtained in the title compound. The product TLC _Rf value: 0.43 (ethyl acetate: hexane = 8: 2).

h) [4S- (4a, 7a, 9a, 9a)] - 4 - {[(Benzyloxy) carbonyl] amino} -5-oxooctahydropyrrolo [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

Dissolve 2.16 g (4.88 mmol) of the compound obtained in (g) above in 50 ml of methylene chloride and add 620 mg of Amberlyst® 15 ion exchange resin, successively with 6 M hydrochloric acid, water, tetrahydrofuran and finally methylene chloride. We washed. The reaction mixture was stirred at room temperature for 3 hours, then filtered and evaporated

The residue was purified by chromatography on a 5 x 20 cm silica gel column eluting with ethyl acetate / hexane (6: 4). As a white foam, 1.34 g of the title product having the thin layer chromatography _Rf value: 0.51 (ethyl acetate: hexane = 8: 2).

i) [4S- (4a, 7a, 9a, 9a)] - 4-Amino-5-axo-octahydro-pyrrolo [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester

The compound prepared as in (h) (1.20 g, 3.17 mmol) is evaporated to dryness three times in succession with toluene and the residue is dried under vacuum overnight and then dissolved in 40 ml of anhydrous methylene chloride the next day. To the solution was added 632 μ; (4.44 mmol, 1.4 equivalents) of iodine trimethylsilane, stirred for 1.5 hours at room temperature under argon and quenched with water (50 mL). The aqueous phase was acidified to pH 1 with 5 ml of 10% hydrochloric acid, washed with 50 ml of ethyl acetate, then basified with 10% sodium hydroxide solution and extracted three times with dichloromethane. The combined extracts were dried over sodium sulfate, filtered, concentrated and the solvent was removed in vacuo to remove the residue. The title compound was obtained as a clear oil (396 mg). The product TLC _Rf value: 0.10 (10% methanol in methylene chloride).

(From the product {4S- [4a (R *), 7a, 9aP]} - 4 - [(3-Phenyl-2-mercaptopropionyl) amino] -5-oxooctahydropyrrolo [2,1b] [1,3] thiazepine-7-carboxylic acid was prepared and its structure was confirmed.

Example 7 {4S- [4a, 7a, 10a] 4-Amino-5-oxooctahydro [1,4] oxazino [3,4-b] [1,3] oxazepine-7-carboxylic acid methyl ester

a) Alld- (2,2,2-trichloroacetimidate)

80% Sodium hydride (945 mg, 31.5 mmol) was washed with hexane (2 x 25 mL) and suspended in anhydrous diethyl ether (30 mL). To this suspension was added dropwise a solution of allyl alcohol (21.4 mL, 18.3 g, 315 mmol) in anhydrous diethyl ether (45 mL), then stirred for 20 min under argon and cooled to 0 ° C with an ice bath. Trichloroacetonitrile (30 mL, 42.3 g, 0.30 mol) was added over about 15 minutes, and the resulting brownish solution was stirred for 40 minutes at 0 ° C, 10 minutes at 10 ° C, and 10 minutes at room temperature. After the reaction time, the mixture was concentrated to a syrup, vigorously stirred with methanol (1.2 mL) and pentane (30 mL) for 5 minutes, the resulting light brown precipitate was filtered off and washed with pentane (2 x 30 mL). The filtrate and washings were combined, concentrated again, then 30 ml of pentane was added to the remaining light brown liquid, stirred for a few minutes, and the resulting suspension was filtered and the precipitate washed with 30 ml of pentane. This operation was repeated at least once, and finally the clear filtrate was evaporated to dryness in vacuo to give 54.0 g of the title compound as a pale red liquid. The product is stored in hexane solution at 10 ° C.

b) N-Phthaloyl-L-serine methyl ester g (161 mmol) of L-serine methyl ester hydrochloride was suspended in 350 ml of water, diluted with 250 ml of dioxane, then added to the resulting clear solution (17 g). 1.0 eq.) Of solid sodium carbonate followed by 37 g (1.05 eq.) Of N-ethoxycarbonylphthalimide. After stirring at room temperature under argon for 2.5 hours, the reaction mixture was extracted with ethyl acetate (3 x 500 mL), and the combined organic extracts were successively washed with 5% sodium bicarbonate (250 mL), The reaction mixture was washed with potassium bisulfate solution and 250 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. The crude residue was purified by chromatography on silica gel (Merck) eluting the column with ethyl acetate / hexane (1: 3) and 1: 2. The appropriate fractions were combined, the solvent was evaporated and the residue was dried in vacuo. The value of 31 g of a thick gum thus obtained the title compound as would TLC _Rf: 0.52 (ethyl acetate: hexane = 1: 1).

c) O-Allyl-N-phthaloyl-L-serine methyl ester

Dissolve 7.37 g (29.5 mmol) of the product from b) above in 30 mL of dichloromethane and add 11.97 g (59.1 mmol; 2 equivalents) of the compound prepared in a) above. ml of cyclohexane solution. Trifluoromethanesulfonic acid (0.37 mL) was then added and after stirring for 20 minutes at room temperature under argon, the precipitate was filtered off and washed with very little dichloromethane. The filtrate and wash were combined, washed with 5% sodium bicarbonate solution (30 mL) and water (30 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. The crude residue was purified by chromatography on silica gel (Merck) eluting with 1: 9 ethyl acetate: hexane. The appropriate fractions were combined and the solvent evaporated in vacuo to give 7.56 g of the title compound as a clear dense syrup. The product TLC _Rf value: 0.70 (ethyl acetate: hexane = 1: 1).

d) O-Formylmethyl-N-phthaloyl-L-serine methyl ester

A solution of 2.5 g (8.64 mmol) of the compound prepared in (c) above in a mixture of 46.4 ml of anhydrous dichloromethane and 4.6 ml of methanol is cooled to -78 ° C with an acetone dry ice bath, followed by ozone. pass through the solution until the blue color persists for about 15 minutes. Nitrogen gas was bubbled through the solution for 10 minutes to remove the blue color, then 14.0 mL (0.19 mol; 22.1 equivalents) of dimethyl sulfide was added and the mixture allowed to warm to room temperature and

Stir under nitrogen for 2.5 hours. After the reaction time, evaporate to dryness and dissolve the remaining honey in 50 ml of ethyl acetate.

After washing with 15 ml of water and 15 ml of saturated sodium chloride solution, the organic phase is dried over anhydrous magnesium sulfate, filtered and the solvent is evaporated off under vacuum. The crude product was purified by chromatography on silica gel (Merck) eluting the column with ethyl acetate: hexane (1: 9.1: 4) and 1: 2. Thus 1.54 g of the title compound is obtained, TLC _Rf value: 0 33 (ethyl acetate: hexane = 1: 1).

e) O- (2,2-Dimethoxyethyl) -N-phthaloyl-L-serine methyl ester

A solution of 1.54 g (5.29 mmol) of the compound prepared as in (d) above in a mixture of 8.3 mL of anhydrous dichloromethane and 8.3 mL of anhydrous methanol is treated with 0.84 mL (7.68 mmol; After adding 45 equivalents of trimethylorthoformate and 92 mg of p-toluenesulfonic acid water (1/1), the solution was stirred at room temperature under argon for 2.5 hours. After the reaction time, the mixture was partitioned between ethyl acetate (50 mL) and saturated sodium bicarbonate solution (15 mL), then the organic layer was washed with water (15 mL) and brine (15 mL), dried over anhydrous sodium sulfate, filtered and evaporate to dryness in vacuo. The crude residue was purified by chromatography on silica gel (Merck) eluting with 1: 4 ethyl acetate: hexane. Value of 1.35 g of a clear, thick syrup thus obtained the title compound, the product TLC _Rf: 0.58 (ethyl acetate: hexane = 1: 1).

en O- (2,2-Dimethoxyethyl) -L-serine methyl ester

Dissolve 2.0 g (5.93 mmol) of the product obtained in (e) above in 14 ml of anhydrous methanol, add 0.30 ml (6.1 mmol) of hydrazine hydrate and then under argon for 4 days at room temperature. stirring. The resulting suspension was filtered, the precipitate was washed with methanol (2 x 14 mL) and the filtrate evaporated to dryness in vacuo. The remaining syrup was redissolved in dichloromethane and the solution was filtered twice before no more precipitate formed. The clear filtrate was evaporated to give 1.17 g of the title compound. The product was a pale yellow syrup with TLC Rf 0.54 (CH 2 Cl 2: MeOH 9: 1).

g) N - [(Benzyloxy) carbonyl] -O - [(tert-butyl) dimethylsilyl] -L-homomer

N - [(Benzyloxy) carbonyl] -L-homoserine (3.0 g, 11.86 mmol) prepared as in Example 6 (a) was dissolved in 65 ml of anhydrous N, N-dimethylformamide and 10.72 g (71.1 mmol) of tert-butyl chlorodimethylsilane and 9.65 g (0.14 mol) of imidazole are added. The reaction mixture was stirred at room temperature under argon for 24 hours, then diluted with methanol (207 mL), allowed to stir at room temperature for another 24 hours and finally concentrated. The residual syrup was dissolved in ethyl acetate (200 mL), washed successively with 10% citric acid (75 mL) and brine (2 x 75 mL), dried (Na 2 SO 4), filtered and the solvent evaporated in vacuo. The crude product was purified by chromatography on silica gel (Merck), eluting the column with ethyl acetate: hexane (1: 1) followed by ethyl acetate: acetic acid (99.5: 0.5). The appropriate fractions were combined, evaporated and the residue was distilled several times with toluene. Value of 3.56 g of a waxy solid thus obtained the title compound, the product TLC _Rf: 0.82 (ethyl acetate: acetic acid = 95: 5).

h) N- {N - [(Benzyloxy) carbonyl] -O - [(tert-butyl) dimethylsilyl] -L-homoseryl} -O- (2,2-dimethoxyethyl) -Serlin-methyl- ester

Dissolve 2.18 g (5.93 mmol) of the product from g) above in anhydrous CH 2 Cl 2 and cool the solution to 0 ° C in a brine bath at this temperature and add 1.71 (5.65 mmol). a solution of the compound prepared in (f) above in 5 ml of anhydrous dichloromethane followed by 0.78 ml (5.65 mmol) of triethylamine and 2.63 g (6.0 mmol) of [(benzotriazol-1-yl) ) oxy] tris (dimethylamino) phosphonium (hexafluorophosphate). The reaction mixture was stirred for 30 minutes at 0 ° C and then 45 minutes at room temperature, then partitioned between diethyl ether (2 x 100 ml) and water (30 ml). The combined organic solvent extracts were washed with 20 ml of 50% saturated sodium bicarbonate solution and 25 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. The crude residue was purified by chromatography on silica gel (Merck) eluting the column with ethyl acetate: hexane (1: 4.1: 3) and 1: 1. Thus 2.38 g of the title product having the thin layer chromatography _Rf value: 0.40 (ethyl acetate: hexane = l: 1).

i) [4S- (4a, 7a, 10a 'S)] - 4 - ([(Benzyloxy) carbamyl] amino} -5-oxooctahydro [1,4-b] oxazino [3,4- b] [1,3] Oxazepine-7-carboxylic acid methyl ester, dissolved in 1.0 g (1.8 mmol) of anhydrous methylene dichloride and dissolved in hydrogen ion phase. Amberlysf ⁸ '15 ion exchange resin and 0.1 ml methanol were added. After stirring at room temperature under argon for 3 days, the resin was filtered off and washed with a little methylene chloride. The filtrate was evaporated and the crude syrup was purified by chromatography on silica gel eluting with 1: 1 ethyl acetate: hexane. The product thus obtained is the title compound, weighing 339 mg, TLC _Rf: 0.53 (ethyl acetate: hexane = 3: 1).

j) [4S- (4a, 7a, 10a (i)] - 4-Amino-5-oxo-octahydro [1,4] -oxazino [3,4-b] [1,3] oxazepine-7-carboxylic acid; methyl ester

771 mg (2.04 mmol) of the compound prepared as in (i) above are dissolved in 25 ml of anhydrous methanol, 125 mg of 10% palladium on charcoal are added and the mixture is hydrogenated at room temperature for 16 hours. After the reaction time, the mixture was filtered through a pad of Celite and the filter was washed with methanol (2 x 25 mL). It's clear

Concentration of the filtrate in vacuo to dryness in vacuo gave 448 mg of the title compound. The value of the product as a syrup TLC _Rf: 0.22 (dichloromethane: methanol = 9: 1).

[From the product {4S- [4a (R *), 7a, 10aP]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydro- [1,4] via oxazino [3,4-b] [1,3] oxazepine-7-carboxylic acid methyl ester {4S- [4a (R *), 7a, 10aP]} - 4 - [(3-phenyl-2-mercapto-propionyl) ) -amino] -5-oxooctahydro- [1,4] -oxazino [3,4-b] [1,3] oxazepine-7-carboxylic acid was identified and identified by elemental analysis.]

Example 8 [4S- (4a, 7a, 10a ')] - 4-Amino-5-oxo-actahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

The product of Example 3 can also be prepared as follows:

a) N - [(Benzyloxy) carbonyl] -O - [(tert-butyl) dimethylsilyl] -L-homoserine

41.56 mmol of N - [(benzyloxy) carbonyl] -L-homoserine prepared as in Example 6 (a) is dissolved in 125 ml of Ν, Ν-dimethylformamide and 37.5 g of Tert-butyl chlorodimethylsilane (249 mmol) was added followed by imidazole (33.95 g, 498 mmol). The resulting pale yellow solution was stirred at room temperature for 22 hours, then diluted with 500 ml of methanol and stirred for an additional 6 hours. Subsequently, methanol and most of the N, N-dimethylformamide are distilled off in vacuo, the residue is taken up in 800 ml of ethyl acetate and washed twice with 300 ml of 10% citric acid solution. The aqueous washings were combined and extracted with ethyl acetate (300 mL), and the combined ethyl acetate layers were washed with water and brine, dried over sodium sulfate and evaporated. The residue was purified by distillation of hexane, and the resulting white powder was dried in vacuo to give 12.942 g of the title compound.

b) [S- (R *, R * j] -2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4 - {[(tert-butyl) dimethylsilyl] oxy} butyryl] amino} -6,6-dimethoxy-hexanoic acid methyl ester

The product (a) (22.78 g, 61.97 mmol) was dissolved in dichloromethane (100 mL) and then N-methylmorpholine (6.81 mL, 61.97 mmol) was added to the solution cooled to 0 ° C. 8.37 g (61.97 mmol) of 1-hydroxybenzotriazole, 10.6 g (51.64 mmol) of (S) -2-amino-6,6, prepared as in Example 1 (e). Dimethoxy hexanoic acid methyl ester was dissolved in methylene chloride (50 mL) and 1- [3- (dimethylamino) propyl] -3-ethylcarbodiimide (11.88 g, 61.97 mmol) was added. The reaction mixture was stirred at 0 ° C for 1 hour, then at room temperature overnight and then concentrated in vacuo. The residue is taken up in 600 ml of ethyl acetate and washed successively with 200 ml of 5% potassium bisulphate solution, 200 ml of 0.5 M sodium hydroxide solution, water and saturated sodium chloride solution. dried over sodium sulfate, filtered and the solvent was evaporated. The residue was triturated with diethyl ether (150 mL), the resulting slurry was filtered and the solid was washed thoroughly with diethyl ether. The filtrate was evaporated to dryness to give 30 g of the title product as a vacuum, an oil, and thin layer chromatography _Rf value: 0.55 (ethyl acetate: hexane = 8: 2). The crude product was used in the next step without further purification.

c) [S- (R *, R *)] - 2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4-hydroxybutyryl] amino} -6,6-dimethoxyhexanoic acid Methyl ester The crude product obtained in (b) above was dissolved in methanol (150 ml), cooled to 0 ° C and stirred at this temperature after addition of 1.96 g of p-toluenesulfonic acid water (1/1). hours. After the reaction time, dissolved in 100 ml of water, 1.3 g of sodium bicarbonate was added, the mixture was concentrated in vacuo and the residue was partitioned between 400 ml of ethyl acetate and 150 ml of water. The aqueous phase was separated and extracted with ethyl acetate (2 x 150 mL), and the ethyl acetate solutions were combined and washed successively with 10% sodium bicarbonate solution, twice with brine, and dried over sodium sulfate. strain finally evaporate to dryness. The crude product residue was purified by flash chromatography on a silica gel column (10 x 25 cm) eluting with 5000 ml of a mixture of 80% ethyl acetate and 20% hexane, 3000 ml of ethyl acetate and 5000 ml of 2% methanol in ethyl acetate. the column. The appropriate fractions were combined and the solvent evaporated in vacuo. The value thus obtained 17.45 g of a pale yellow oil of the title compound, the product TLC _Rf: 0.17 (ethyl acetate: hexane = 8: 2).

d) [S- (R *, R *)] - 2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4 - {[methylsulfonyl] oxy} butyryl] amino} -6,6-dimethoxy-hexanoic acid methyl ester

The compound (17.40 g, 39.50 mmol) obtained as described above under (c) is dissolved three times in succession in toluene, and the solution is always concentrated, and the residue is then dried in vacuo overnight. The following day, the completely dry starting material thus obtained was dissolved in 250 ml of anhydrous dichloromethane, cooled to -15 ° C with ice-acetone, and then freshly distilled triethylamine (8.26 ml, 59.28 mmol) was added. followed by dropwise addition of 3.67 mL (47.4 mmol) of methanesulfonyl chloride. The reaction mixture was stirred for 30 minutes at -15 ° C, then quenched with 100 mL of saturated ammonium chloride solution. After stirring for 5 minutes, the mixture was diluted with ethyl acetate (600 mL), potassium bisulfate (5%) and brine, dried over sodium sulfate and evaporated to dryness. The residue was dried in vacuo to give 20.40 g of the title compound. The product, a yellow oil, and thin layer chromatography _Rf value: 0.35 (ethyl acetate: hexane = 8: 2), used in the next step without further purification.

e) [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2 - {[(benzyloxy) carbonyl] amino} butyryl] amino} -6.6 dimethoxy-hexanoic acid methyl ester

HU 217 960 Β

Thioacetic acid (5.09 mL, 71.10 mmol) was dissolved in methanol (100 mL) followed by cesium carbonate (10.81 g, 33.18 mmol), and the solution was concentrated in vacuo. The resulting solid was triturated three times with anhydrous acetone and the resulting cesium thioacetate was dried over phosphorus (V) oxide overnight.

Dissolve 20.40 g (39.50 mmol) of the compound prepared as in (d) above in 150 ml of anhydrous Ν, Ν-dimethylformamide and then add 10.576 g (50.85 mmol) of cesium thioacetate in a syringe. ml of suspension in ml, Ν-dimethylformamide. The resulting yellow solution was stirred overnight at room temperature under argon and then evaporated under high vacuum the next day to remove most of the N, N-dimethylformamide. The residue was taken up in 1000 ml of ethyl acetate and washed with 10% sodium bicarbonate solution (200 ml) and saturated sodium chloride solution (4 x 200 ml), dried over sodium sulfate, filtered and the solvent was evaporated. The residual crude product was distilled three times successively with toluene and dried in vacuo to give 20 g of the title compound. The yellow solid product with an _Rf value of TLC: 0.47 (ethyl acetate: hexane = 8: 2), used in the next step without further purification.

f) [S- (R *, R *)] - 2 - {[2 - {[(Benzyloxy) carbonyl] amino} -4-mercaptobutyl] amino} -6,6-dimethoxyhexanoic acid of methyl ester (39.50 mmol) in a solution of the product obtained in (e) above in 250 ml of methanol and cooled to 0 ° C was purged with argon gas for 15 minutes. Thereafter, 9.17 mL (40 mmol) of 25% (w / w) 0.945 in methanolic sodium methylate solution was added dropwise with continuous argon gas. After stirring for 5 minutes, the reaction mixture was quenched with saturated ammonium chloride solution (200 mL) and shaken well with 1000 mL of ethyl acetate and 200 mL of water. The aqueous phase was further extracted with 200 mL of ethyl acetate, and the combined ethyl acetate extracts were washed with 400 mL of saturated ammonium chloride solution and 400 mL of saturated sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. The title compound was obtained as a yellow oil (17.5 g). The product having a thin layer chromatography _Rf value: 0.45 (ethyl acetate: hexane = 8: 2), used in the next step without further purification.

g) [4S- (4a, 7a, 10a ')] - 4 - {[(Benzyloxy) carbonyl] amino} -5-oxooctahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester

17.5 g (38.3 mmol) of the compound prepared as in f) above are dissolved in 600 ml of dichloromethane and 6 g of the appropriately prepared Amberlyst® 15 ion exchange resin, first with 6 M hydrochloric acid and then with water, tetrahydrofuran, are added. and finally washed with dichloromethane and then dried. After the resin was added, the suspension was stirred under argon at room temperature for 18 hours, then filtered, the filtrate was evaporated and the residue was adsorbed onto Celite. The crude product was purified on a 10 x 30 cm silica gel column eluting with 20-30% ethyl acetate / hexane. The appropriate fractions were combined and the solvent evaporated in vacuo. The title compound was obtained as a yellow oil (9.18 g), TLC Rf 0.32 (ethyl acetate: hexane = 1: 1).

h) [4S- (4a, 7o, 10a ')] -4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

9.1 g (23.19 mmol) of the compound prepared as in (g) above are weighed, concentrated three times in succession with toluene and finally dried under vacuum overnight. The thus-dried starting material was dissolved in dichloromethane (150 mL), followed by the dropwise addition of iodo-trimethylsilane (4.95 mL, 34.78 mmol). The resulting yellow solution was stirred under argon at room temperature for 1.5 hours and then quenched with 120 mL of 0.4 M HCl in methanol / dioxane. The mixture was concentrated in vacuo and the residue was partitioned between diethyl ether (500 mL) and water (700 mL). The organic layer was extracted with additional 150 mL of 0.1 M hydrochloric acid, then the combined aqueous layer was cooled to 0 ° C and at this temperature, the pH of the solution was monitored by pH meter with 1 M sodium hydroxide solution to pH 10.5. basic. The basic solution was extracted with dichloromethane (4 x 400 mL), and the combined organic extracts were washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo to give 6.95 g of the title compound as a yellow oil. TLC Rf = 0.20 (methanol: dichloromethane = 1: 9).

(The product {4R- [4ct (R *), 7a, 10aP]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydro-7H-pyrido [2 , 1b] [1,3] Thiazepine-7-carboxylic acid methyl ester {4S- [4a (R *), 7a, 10aP]} - 4 - [(3-phenyl-2-mercaptopropionyl) -amine ] -5-oxooctahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid identified by elemental analysis.)

Example 9 [4S- (4a, 7a, 10a)] - 4-Amino-5-oxo-octahydro [1,4] oxazino [3,4-b] thiazepine-7-carboxylic acid methyl ester

a) N- {N - [(Benzyloxy) carbonyl] -L-homoseryl} -O (2,2-dimethoxyethyl) -L-serine methyl ester

5.56 g (10 mmol) of N- {N - [(benzyloxy) carbonyl] -O - [(tert-butyl) dimethylsilyl] -L-homomoseryl} prepared as in Example 7 (h). -O- (2,2-Dimethoxyethyl) -L-serine methyl ester was dissolved in methanol (65 ml), cooled to 0 ° C in a brine bath, and then p-toluenesulfonic acid water (386 mg, 2.0 mmol) was added. (1/1) and stirred at 0 ° C for 1.5 h. At the end of the reaction time, dissolved in 20 ml of water, 198 mg of sodium bicarbonate was added and after 5 minutes of stirring the methanol was evaporated.

HU 217 960 Β

The aqueous residue was extracted with ethyl acetate (2 x 200 mL), and the combined organic extracts were washed with water (110 mL), 5% sodium bicarbonate (80 mL), and brine (80 mL), and dried over anhydrous sodium sulfate. , finally filter under vacuum to dryness. The crude product was purified by column chromatography over silica gel (Merck), eluting with ethyl acetate: hexane (2: 1) and then ethyl acetate: methanol (98: 2). In this way, the title compound is obtained in the form of a syrup (3.975 g), TLC 0.17 (ethyl acetate: hexane = 4: 1).

b) N- {N - [(Benzyloxy) carbonyl] -O- (methylsulphonyl) -L-homoserryl} -O- (2,2-dimethoxyethyl) -L-serine methyl ester

A solution of 3.975 g (8.98 mmol) of the compound prepared in (a) above in 52 mL of anhydrous dichloromethane is cooled to -15 [deg.] C. and then 1.82 mL (13.1 mmol) of triethylamine and methanesulfonyl chloride (0.82 mL, 10.6 mmol). The reaction mixture was stirred for 30 minutes at -15 ° C and then quenched with 19 ml of 25% ammonium chloride solution. The mixture was allowed to warm to room temperature, diluted with ethyl acetate (750 mL), and the organic layer was separated with 100 mL of 5% potassium bisulfate, 100 mL of semi-saturated sodium chloride, and 100 mL of saturated sodium chloride. solution, dried over anhydrous sodium sulfate, filtered and finally evaporated to dryness in vacuo. The residue is 4.9 g of waxy product having a thin layer chromatography _Rf: 0.32 (ethyl acetate: hexane = 4: 1) to give the title compound.

c) N- {S-Acetyl-N - [(benzyloxy) carbonyl] -L-homocysteinyl} -O- (2,2-dimethoxyethyl) -L-serine methyl ester

To a solution of 2.6 ml of thioacetic acid in 40 ml of anhydrous methanol was added 5.56 g (17.04 mmol) of cesium carbonate and, after stirring for 10 minutes, the solution was evaporated to dryness. The solid residue was triturated with acetone (8-8 mL) seven times and the off-white product dried in vacuo. This gives 4.39 g of cesium thioacetate, which is the cesium salt of thioacetic acid.

2.43 g (1.3 equivalents) of the above cesium thioacetate are suspended in 8.0 ml of anhydrous Ν, Ν-dimethylformamide and 4.9 g (8.98 mmol) of the compound prepared as in b) above are added. of anhydrous Ν, Ν-dimethylformamide and stirred at room temperature for 16 hours, then diluted with 1000 ml of ethyl acetate and successively with 2 x 150 ml of 5% sodium bicarbonate solution, 2 x 150 ml of water. and washed with 150 ml of saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo to give a crude residue which was purified by chromatography on silica gel (Merck) eluting with 1: 1 and 2: 1 ethyl acetate / hexane. The resulting waxy solid (3.93 g) is the title compound; TLC _Rf value: 0.63 (ethyl acetate: hexane = 4: 1).

d) N- [N - [(Benzyloxy) carbonyl] -L-homocysteinyl} O- (2,2-dimethoxyethyl) -L-serine methyl ester

200 mg (0.49 mmol) of the compound prepared as in (c) above are dissolved in methanol, purged with argon gas for 30 minutes and then cooled to 0 ° C in a salt-ice bath and continued during the addition and reaction. argon gas was bubbled through a solution of 0.11 mL (0.5 mmol) of 25% sodium methoxide in methanol. After 5 minutes at 0 ° C, 2.3 ml of 25% ammonium chloride solution was added, then partitioned between 12 ml of ethyl acetate and 2.3 ml of water. The combined organic solvent extracts were washed with 4.6 mL of 25% ammonium chloride solution and 9.6 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. Value of 183.2 mg of a white solid was the title compound, the product TLC _Rf: 0.62 (ethyl acetate: hexane = 4: 1).

e) [4S- (4a, 7a, 10a)] - 4 - {[(Benzyloxy) carbonyl] amino} -5-oxooctahydro [1,4] oxazino] 3,4] [1, 3] Thiazepine-7-carboxylic acid mg (0.11 mmol) of the product obtained as in (d) above was dissolved in 2.0 ml of anhydrous methylene chloride after addition of 13 mg of the Amberlyst® 15 ion exchange resin in hydrogen phase for 3 days. under argon at room temperature. At this time, another 13 mg of Amberlyst® 15 ion exchange resin was added to the solution, stirring continued for a further 3 days, and the solution was decanted and purified by chromatography on a silica gel column. The column was eluted with ethyl acetate / hexane (1: 3) to give 21.1 mg of the title compound. The value of the product as a syrup, TLC _Rf: 0.70 (ethyl acetate: hexane = 4: 1).

j) [4S- (4a, 7a, 10a ')] - 4-Amino-5-oxo-octahydro [1,4] oxazino [3,4-b] [1,3] thiazepine-7-carboxylic acid methyl ester

421 mg (1.01 mmol) of the compound prepared as in (e) above are dissolved in 25 ml of anhydrous dichloromethane, and 0.72 ml (5.06 mmol) of iodtrimethylsilane are added and the solution is stirred at room temperature under argon. For 75 hours. After the reaction time, the mixture was evaporated to dryness and the residual syrup was partitioned between diethyl ether (50 mL) and 0.2 M hydrochloric acid (2 x 25 mL). The aqueous phase was adjusted to pH 10 with 25 mL of saturated sodium bicarbonate solution, 2.0 g of solid sodium chloride was added, and then extracted with 3 x 75 mL of dichloromethane to give 219 mg of the title compound. in the form of a dense honey-like residue. An additional 2.0 g of sodium chloride was added to the aqueous phase, which was then re-extracted with dichloromethane (2 x 100 mL) to give an additional 37 mg of the title compound. The product TLC _Rf value: 0.23 (methylene chloride: methanol = 9: 1).

EN 217 960 Β (The product {4S- [4a (R *), 7a, 10aP]} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydro [1 , 4-b] oxazino [3,4-b] [1,3] thiazepine-7-carboxylic acid methyl ester {4S- [4a (R *), 7a, 10ap]} - 4 - [(3- phenyl-2-mercapto-propionyl) -amino] -5-oxo-octahydro [1,4] oxazino [3,4-b] [1,3] thiazepine-7-carboxylic acid, identified by elemental analysis.)

Example 10 [4S- (4a, 7a, 10a $)] - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester (p-toluenesulfonate) )

6.11 g [4S- (4oc, 7a, 10a3)] - 4-Amino-5-oxooctahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester Dissolve in 100 ml of ethyl acetate and add a solution of 4.52 g of p-toluenesulfonic acid water (1/1) in a mixture of 3 ml of methanol and 20 ml of ethyl acetate. A precipitate precipitated immediately from the solution. The mixture was diluted with additional ethyl acetate, then the precipitate was filtered off, washed with diethyl ether and finally dried in vacuo. The title compound was obtained as a pale yellow crystalline product, m.p. 179-181 ° C (98% purity).

Example 11 [4S- (4a, 7a, 10a ')] - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

In addition to the Examples 3 and 8, the title compound can be prepared by the following procedure:

o) N - [(Benzyloxy) carbonyl] -L-methionyl

Into a 2 liter flask equipped with a mechanical stirrer and a thermometer suitable for measuring the internal temperature, 61.65 g (1.541 mol) of sodium hydroxide are weighed and dissolved in 1000 ml of distilled water. To the solution was added L-methionine (100.0 g, 0.670 mole) at room temperature, an ice bath was placed under the flask, and when the internal temperature had dropped to 3 ° C, 110 ml (0.737 mole) of benzyl chloro was added over 10 minutes. formate) respectively. After an induction period of about 15 minutes, the internal temperature rises from 3 ° C to 12 ° C over 30 minutes and then decreases to 0 ° C over a further 15 minutes. The reaction mixture was stirred for 2 hours at 0 ° C, during which time the initially cloudy solution became homogeneous. The ice bath was removed and the mixture allowed to warm to room temperature over about 1 hour, transferred to a separatory funnel and washed with 2 x 300 mL hexane. The aqueous phase was first acidified to pH 5 with 6 M hydrochloric acid, then diluted with 600 mL of ethyl acetate, then further lowered to pH 2. The organic layer was separated, the aqueous layer was extracted with ethyl acetate (3 x 600 mL), and the combined organic extracts were washed with brine (750 mL), dried (MgSO4), filtered and concentrated in vacuo. The crude product was dissolved in 1500 ml of toluene, evaporated, about half of the toluene was evaporated, 750 ml of fresh toluene was added to the residue and evaporation continued until 630 ml of a toluene solution was obtained. This solution was kept at 5 ° C overnight while little product crystallized out of solution but redissolved the next day at room temperature. At this time, 134 mL of toluene was added to the solution while retaining a few seed crystals, and 500 mL of heptane was added each time for 30 minutes with vigorous motor stirring for 10 min. After the addition of the last portion, the solution begins to crystallize. Continue the addition of heptane and the following

Heptane (1020 ml) was added over 1.5 hours, the resulting suspension was stirred for 2 hours, then filtered, the crystals were suctioned off, and washed with 3 x 150 ml of a 1: 2 mixture of toluene and heptane (3 x 500 ml). . 156.8 g of a white crystalline product, m.p. 66 ° C, are obtained; TLC _Rf value: 0.78 (ethanol: water = 3: 1). [α] _D = -1.5 ° (c = 1; 95% ethanol).

b) N - [(Benzyloxy) carbonyl] -L-methionine methyl ester

Into a 3 L mechanical flask fitted with a argon gas inlet flask, 100.0 g (0.353 mol) of the compound prepared as in (a) above are weighed, dissolved in 2000 ml of methanol and 6.71 g (0.035 mol). After adding p-toluenesulfonic acid water (1/1), the solution was stirred under argon for 22 hours. Triethylamine (4.9 mL, 0.035 mol) was added and the mixture was allowed to stir for 15 minutes and then concentrated in vacuo. The residual pale yellow oil was dissolved in 900 ml of ethyl acetate, washed successively with 740 ml of 1M hydrochloric acid, 2 x 740 ml of saturated sodium bicarbonate solution and 740 ml of saturated sodium chloride solution, and the organic phase was washed with magnesium sulfate. dried, filtered and the solvent evaporated in vacuo. The residue is a light yellow oil, to which 100 ml of hexane (100 ml) are added successively and each is evaporated twice to dryness. 98.22 g of the title compound are thus obtained, which is a white solid.

c) N - [(Benzyloxy) carbonyl] -L-methionine methyl ester S-oxide

In a 3 liter flask fitted with a mechanical stirrer, 97.95 g of the product obtained in (b) above are dissolved in a mixture of 1675 ml of methanol and 215 ml of distilled water. While cooling with ice, the solution first

Sodium hydrogen carbonate (28.5 g, 0.339 mol) was added and N-chlorosuccinimide (44.0 g, 0.329 mol) was added in small portions over 25 minutes at a temperature not exceeding 7 ° C. The reaction mixture was stirred for an additional 1 hour in an ice bath, then allowed to warm to room temperature over about 1 hour, then concentrated in vacuo, evaporating about 75% methanol. The residue was diluted with ethyl acetate (1000 mL), washed with brine (500 mL), and the aqueous phase was back-extracted with ethyl acetate (2 x 200 mL). The organic solvent solutions were combined, dried over magnesium sulfate, filtered and the solvent evaporated in vacuo. 100 to 100 ml of toluene are distilled three times in succession from the remaining clear viscous oil.

Finally, the solvent was removed under high vacuum to give the title compound as a clear, transparent oil, which subsequently solidified. The white solid weighs 131.4 g but has 12% by weight of succinimide and 9% by weight of toluene. The product, moreover, being a sulfoxide, is a mixture of diastereomers.

d) S-Acetoxymethyl-N - [(benzyloxy) carbonyl] -L-homocysteine methyl ester

A 1 liter flask is charged with 102.8 g (corrected mass 0.328 mol) of the product obtained in (c) above, 480 ml toluene, 32.3 g (0.394 mol) sodium acetate and 186 ml (1.970 mol) acetic anhydride. The mixture was heated at reflux (118 ° C) for 18 hours under argon and allowed to cool to room temperature. Upon completion of the reaction, the dark brown mixture thickens at room temperature from the precipitated solid. The solid was dissolved in ethyl acetate (100 mL) and concentrated in vacuo to give a brown viscous residue. This residue was evaporated from toluene (240 mL) to remove acetic anhydride, and the residue was dissolved in ethyl acetate (1000 mL) and washed gently with saturated sodium bicarbonate (4 x 680 mL). The organic phase was then shaken with an additional 950 ml of saturated sodium chloride solution, dried over magnesium sulfate, filtered and the solvent evaporated in vacuo. To remove solvent residues, the residue was suctioned under high vacuum for a while, then the light brown solid crude was dissolved in 450 mL of butyl acetate with heating (35 ° C) and stirring. The solution is cooled to room temperature and 200 ml of hexane are added slowly, then allowed to stir for 15 minutes, when crystals begin to separate from the solution. An additional 700 mL of hexane was added over about 30 minutes, the resulting suspension was stirred for 3 hours, and the product was filtered and successively with 200 mL of 1: 2 butyl acetate: hexane, twice 400 mL of 1: 4 butyl acetate. After washing with hexane and twice with 250 ml of hexane, it is first dried in air, then dried under high vacuum. 87.7 g of the title compound are obtained. M.p. 73 ° C; TLC _Rf value: 0.80 (5% methanol in methylene chloride).

[α] _D = -1.6 ° (c = 1; 95% ethanol).

e) S-Acetyl- [N- (benzyloxy) carbonyl] -L-homocysteine

A 1 liter flask was charged with 83.0 g (0.233 mol) of the compound obtained in (d) above, dissolved in 415 ml of tetrahydrofuran, and then bubbled with argon for 30 minutes. In another 2-liter flask equipped with a motor stirrer and argon gas inlet, a solution of 62.7 g (0.969 mol) of 86.8% potassium hydroxide and 280 ml of distilled water is made, and argon gas is bubbled through this solution for 15 minutes. . Subsequently, to the potassium hydroxide solution at an internal temperature of 20 [deg.] C., the tetrahydrofuran solution is added rapidly under argon, with vigorous stirring, and the compound of (d) is rinsed with 20 ml of tetrahydrofuran previously purged with argon for 15 minutes. After 30 minutes, a clear biphasic mixture was obtained while a slight exothermic reaction occurred, as the reaction mixture was heated to 28 ° C. After a further 2 hours, the mixture was cooled to 1 ° C (internal temperature) and 2.75 g (0.073 mol) of sodium tetrahydroborate was added in one portion. Once again, an exoteimic reaction occurs, raising the temperature to 6.8 ° C, but after 20 minutes the mixture cools back to 0 ° C. At this time, the temperature was allowed to rise to 11 ° C over 30 minutes, and the mixture was again cooled to 1 ° C and 68.6 ml (0.727 mol) of acetic anhydride was added over 10 minutes. The addition is followed by warming again, with a maximum temperature of about 10 ° C, but drops again to 4 ° C by the end of the addition. The cooling bath was then removed and the reaction mixture was allowed to stir at room temperature for 45 minutes and then concentrated in vacuo to about half the original volume. The residue was acidified to pH 2 with 175 mL of 6 M hydrochloric acid, extracted twice with successively 1100 mL of 1100 mL of ethyl acetate, and then the combined organic solvent extracts were washed with 560 mL of saturated sodium chloride solution, treated with charcoal and glacial magnesium sulfate. filtered and finally concentrated in vacuo. A yellow oil remained, to which 380 ml of butyl acetate was added and the solution was concentrated at 95 ° C to about half its volume. Fresh butyl acetate (190 mL) was added to the residue, and the solvent was evaporated again to leave 190 mL of butyl acetate, then slowly added heptane (300 mL) with vigorous stirring and a few seed crystals as the solution opalescent. we add. As a result, a white crystalline precipitate appears after 15 minutes. An additional 570 mL of heptane was added over the next 30 minutes, the resulting suspension was allowed to stir overnight at room temperature, the next day the crystalline product was filtered and washed with 2 x 275 mL 1: 3 butyl acetate: heptane and 2 x 275 mL hexane. , then dried under high vacuum. The title compound was obtained as a white crystalline solid (50.1 g), m.p. 73-74 ° C. TLC Rf 0.83 (ethanol: water = 3: 1).

[α] _D = -1.30 (c = 1; 95% ethanol).

The filtrate was concentrated to about 100 mL, then heptane (310 mL) was added to the residue as above to give 8.4 g of the second generation product. This is also a white crystalline material, identical to the title compound, giving a total yield of 58.5 g.

en [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2 - {[(benzyloxy) carbonyl] amino} butyryl] amino} -6.6 acid, methyl ester dimethoxysilane

S-acetyl-N - [(benzyloxy) carbonyl] -L-homocysteine (0.456 mol) was dissolved in methylene chloride (600 ml) and Ν, Ν-dimethylformamide (90 ml), 1-hydroxy (64.72 g, 0.479 mol). After addition of benzotriazole water (1/1), the solution was cooled in an ice bath and then 93.7 g was added.

A solution of (S) -2-amino-6,6-dimethoxy-hexanoic acid methyl ester prepared as described in Example 1 (e) in 600 ml of dichloromethane. Subsequently, 91.83 g (0.479 mole) of 1- [3- (dimethylamino) propyl] -3-ethylcarbodiimide hydrochloride was added, which was then stirred at 0 ° C for 1 hour and then for 2 hours. Allow to stir at room temperature. After the reaction time, the mixture was concentrated in vacuo and the residue was partitioned between ethyl acetate (3000 mL) and saturated aqueous sodium bicarbonate solution (1000 mL). The organic layer was washed with water (1000 mL), 5% potassium bisulfate (1000 mL), water (1000 mL), and brine (1000 mL), dried (Na 2 SO 4) and the solvent was evaporated in vacuo. The crude product (238 g) was dissolved in ethyl acetate: methylene chloride (1: 1, 300 mL), applied to a 10 x 15 cm silica gel (Merck) filter bed, and then silica gel (7000 mL, 8: 2 ethyl acetate: hexane). and 4,000 mL of ethyl acetate. 205.28 g of the title compound are obtained.

g) [4S- (4a, 7a, 10aft)] - 4 - [[(Benzyloxy) carbonyl] amino} -5-oxooctahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester

The compound (205.28 g, 0.412 mol) previously azeotroped with dichloromethane and toluene and thus completely dehydrated was prepared as described in (f) above in methanol (2000 ml) and cooled to 0 ° C with an ice bath. Argon gas was passed through it for 30 minutes. Subsequently, while purging with argon gas, 95.1 ml (1.01 equivalents) of 25% (w / w) sodium methoxide in methanol was added rapidly, followed by stirring for a further 10 minutes, followed by 1000 ml of saturated ammonium chloride. solution is quenched. Water (500 mL) and ethyl acetate (3000 mL) were then added and the mixture was partitioned in vacuo, evaporating the organic solvents, ethyl acetate and methanol. Fresh ethyl acetate (1000 ml) was added to the remaining aqueous portion, the immiscible layers were separated, and the organic layer was washed with saturated ammonium chloride solution (500 ml), and the combined aqueous solution was back-extracted with ethyl acetate (1000 ml). The combined organic phases are then washed with 1000 ml of water and twice with 1000 ml of saturated sodium chloride solution, dried over sodium sulphate, filtered and evaporated. Methylene chloride (dichloromethane) was added to the residue, evaporated again and dried in vacuo to give 182.65 g of intermediate (f) but containing free mercapto group.

0.400 mol of the above mercapto compound is dissolved in 4000 ml of dichloromethane, 30.8 ml (0.400 mol) of trifluoroacetic acid are added, the mixture is refluxed for 16 hours, cooled and evaporated in vacuo. The residue was dissolved in 2000 ml of ethyl acetate, washed with 400 ml of 0.1 M hydrochloric acid, 1000 ml of water, 1000 ml of saturated sodium bicarbonate solution, again with 1000 ml of water and finally with 1000 ml of saturated sodium chloride solution. dried over sodium sulfate and the solvent was evaporated. The residue was re-evaporated with dichloromethane and dried in vacuo to give 166.29 g of the title compound.

h) [4S- (4a, 7a, 10a $)] - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester

162.93 g (0.414 mol) of the compound prepared as described above under g) are dissolved in 1500 ml of dichloromethane and then 76.6 ml (0.538 mol) of iodotrimethylsilane are added under argon. The reaction mixture was stirred for 1.5 hours, then concentrated in vacuo and partitioned between ethyl acetate (1000 mL) and 1M hydrochloric acid (700 mL). Addition of acid produces carbon dioxide at pH 1.2. The ethyl acetate layer was separated and extracted with 300 mL of 1M hydrochloric acid, and the aqueous solutions were combined, washed with 1 x 1000 mL of ethyl acetate, then cooled to 0 ° C, and 4M sodium hydroxide solution to pH 10.0. basic. The volume of sodium hydroxide solution is approximately 275 ml. The basic aqueous extract was saturated with solid sodium chloride, extracted with dichloromethane (5 x 1000 mL), the combined organic extracts dried over sodium sulfate, and the solvent was evaporated in vacuo. The residue was redissolved in methylene chloride (1000 mL), washed with brine (500 mL), then filtered and evaporated again. This way

98.8 g of the expected product are obtained.

(The product {4S- (4a (R *), 7a, 10aP)} - 4 - {[2- (acetylthio) -3-phenylpropionyl] amino} -5-oxooctahydro [7H-pyrido [2 , 1-b] [1,3] thiazepine-7-carboxylic acid methyl ester {4S- [4a (R), 7a, 10aP]} - 4 - [(3-phenyl-2-mercaptopropionyl) -amino] Converted to -5-oxo-octahydro [7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid whose structure was confirmed.

Example 12 [4S- (4a, 7a, 10a)] - 4-Amino-5-oxo-octahydro-7H-pyrido [2,1-b] [1,3] thiazepine-7-carboxylic acid methyl ester

The product described in Examples 3, 8 and 11 can also be prepared as follows:

a) Diethyl ester of 2- (acetylamino) -2- (4-acetoxybutyl) malonic acid

Weigh it into a flask filled with argon gas

60.8 g (2.532 mol) of 95% sodium hydride and 500 ml of anhydrous Ν, dimet-dimethylformamide were cooled to 0 ° C in an ice bath, then added with stirring over a period of about 45 minutes. (2.302 moles) of diethyl [(acetylamino) malonate] in 1200 ml of anhydrous Ν, Ν-dimethylformamide while maintaining the temperature of the mixture below 18 ° C. After the addition was complete, the cloudy solution was allowed to gradually warm to room temperature, stirred for an additional 1 hour, then added;

471.5 g (2.417 mol) of (4-bromobutyl) acetate. The mixture was allowed to stir at 59-60 ° C for 18 hours and then cooled to room temperature. Beadago20

40 ml of anhydrous ethanol and 4 ml of anhydrous concentrated acetic acid were added, after stirring for another 15 minutes, the mixture was poured into 10% lithium chloride solution and extracted with 2 x 3,000 ml of ethyl acetate. The combined ethyl acetate extracts were washed with 10% lithium chloride solution (3 x 3000 mL), dried over anhydrous sodium sulfate, and evaporated in vacuo. The residual oil (750 g) is the title compound.

b) 2- (Acetylamino) -6-hydroxyhexanoic acid

Into a 5 liter 3-necked flask fitted with a thermometer, a magnetic stirrer and an air-cooled reflux condenser was charged 730 g (2.2 mol) of the compound prepared in (a) above, then 300 ml of anhydrous ethanol was added, followed by 1600 ml. 6 mol) of 6 M aqueous sodium hydroxide solution. The reaction mixture was heated to 68-70 ° C and maintained at this temperature for 5 hours, resulting in a homogeneous solution. The solution is then cooled to room temperature and the pH is adjusted to 1.3 with 6M hydrochloric acid (approximately 1320 mL) added slowly. The flask is fitted with a short-path distillation apparatus necessary to distill the ethanol out of the system while slowly raising the temperature to 87-90 ° C. The mixture is kept at this temperature for a total of 8.5 hours, which results in a slow evolution of gas, formation and removal of carbon dioxide. At the end of the reaction, the total volume of distillate was 600 ml and the pH of the solution was 3.0. The reaction mixture was concentrated in vacuo and 2 x 500 mL toluene was distilled off to completely remove water. The residual semi-solid mass was triturated with ethanol (1000 mL), filtered, the filtered salt washed with anhydrous ethanol (500 mL), and the filtrate was concentrated in vacuo. In this way, 509 g of crude product, 82% pure, containing ethanol and toluene were obtained as an oil.

c) (S) -2-Amino-6-hydroxyhexanoic acid

443 g of the crude product obtained as described in b) above, containing a small amount of toluene and ethanol, are estimated to be 394 g in weight, dissolved in 3300 ml of water and 1 L of lithium hydroxide is added to bring the pH of the mixture to 7.5 (approximately 1530 ml of lithium hydroxide solution is required). The mixture was then heated to 35 ° C and 0.4 g of acylase (grade 1, isolated from porcine kidney) was allowed to stir for 24 hours. At the end of this period, the pH of the mixture was 7.33. The pH is adjusted to 7.5 with about 2 ml of 1 M lithium hydroxide solution, another 0.4 g of acylase is added, and stirring is continued for another 17 hours, when pH is again measured. . The mixture, which was adjusted to pH 5.9 with acetic acid, was then heated to 92 ° C after the addition of 20 g of Celite and 20 g of charcoal and kept at that temperature for 5 minutes. The mixture was then filtered through a pad of Celite, and the filtrate was concentrated in vacuo and the resulting semi-solid slurry (441 g) was squeezed in 900 mL of water: ethanol: N, N-dimethylformamide (900 mL). Rubbing the rest requires some heating. The suspension was then placed in the refrigerator overnight, filtered the next day, and the filtered product was washed with 200 ml of the above solvent mixture. The crude product (214 g) thus obtained contains approximately 40% of the N-acetyl derivative and is suspended in methanol (500 ml), heated on a steam bath, allowed to stand for 2 hours and filtered. This operation was repeated a second time to give 108 g of the title compound.

[α] _D = + 22 ° (c = 1.44; 6 M hydrochloric acid).

The compounds of (b) and (c) above may also be prepared by the following procedure:

b ') 2- (Acetylamino) -6-hydroxyhexanoic acid

Into a 5 L three-necked flask equipped with a motor stirrer and thermocouple thermometer, 631 g (1.933 mol) of the compound prepared in (a) and 259 ml of tetrahydrofuran were weighed. Subsequently, 1385 ml (8.31 moles) of 6 M sodium hydroxide solution was added with stirring over about 40 minutes, resulting in a highly exothermic reaction, requiring ice cooling to keep the temperature below 60 ° C. After the addition was complete, the mixture was heated to gentle reflux for 5.5 hours while the temperature was 67-68 ° C and allowed to stir at room temperature overnight (16.5 hours total). The next day, 1150 ml (6.9 mol) of 6 M hydrochloric acid was added portionwise to adjust the pH of the mixture from 12.75 to 1.30, maintaining the temperature at about 25 ° C, and then fitting the flask with a short-run distillation apparatus, and gradually raising the temperature until distillation and gas evolution (decarboxylation, carbon dioxide evolution) begins. The cauldron temperature is 72.3 ° C and the head temperature 70 ° C. Increasing the temperature further slows down the gas evolution and stops the distillation. At this point, the boiler temperature reaches 84.1 ° C, but the head temperature drops to 50 ° C. The distillate was 410 ml and the pH of the mixture remaining in the pan was 3.9. Continuing the heating for a further 10 minutes, the gas evolution ceases completely and the elapsed reaction time is exactly 7.5 hours from the start of distillation.

The reaction mixture was allowed to stir again overnight, and the next day, the clear, crisp, pH 3.5 solution was concentrated in vacuo on a rotary evaporator using a 60 ° C bath. Anhydrous ethanol (750 ml) was added to the pasty residue and the resulting crystal suspension was evaporated again. Here again, a rotary evaporator is used, the water bath temperature is 60 ° C, but a better vacuum (pump) is established. 750 ml of anhydrous ethanol are distilled off twice from the residue, and the resulting residue is stirred with 1500 ml of ethanol in a 60 ° C bath until a fine crystalline mass is obtained. This takes about 20 minutes. After stirring for another 20 minutes at room temperature, the suspension is filtered and the filtered salt washed with 2 x 300 ml of anhydrous ethanol. The filtrate is slightly turbid and this turbidity is eliminated by filtering again on a Celite filter bed. The crude clear filtrate was then concentrated on a rotary evaporator to give 434.6 g of the title compound. The product is a golden yellow, dense syrup, a

GB 217,960 Β TLC _Rf value: 0.59 (methanol: acetic acid: water = 10: 1:. 1).

c ') (S) -2-Amino-6-hydroxyhexanoic acid

Into a 5 L three-necked flask equipped with a mechanical stirrer and a thermometer, 434 g (1.93 mol) of the compound prepared as described under b 'above and 3000 ml of water are weighed. The resulting cloudy solution was adjusted to pH 4.05 with 7.5 L of 705 ml of 1 M lithium hydroxide solution and the temperature was raised to exactly 36 ° C, and 0.710 g of porcine kidney acylase 1 was added to the mixture. For 5 hours at 35-36 ° C. After the reaction time, the mixture was cooled to room temperature and the pH adjusted to about 5.9 with 4.4 mL of anhydrous acetic acid (4.4 mL) at the end of the enzyme reaction and 29 g of Celite and 29 g of charcoal were added. . The mixture is heated to 91 ° C with stirring, then the bath is removed to allow it to cool to room temperature and the suspension is filtered through a 18.5 cm filter paper disk and the filter is thoroughly washed with water. About 3900 ml of the colorless filtrate was evaporated on a rotary evaporator at a bath temperature of 60 ° C, and the remaining 476 g of a clear thick oil were mixed with 720 ml of anhydrous ethanol until a homogeneous crystal suspension was formed. The solvent was distilled off again, 1584 ml of anhydrous ethanol were added to the white solid residue, and the suspension was rotated overnight at room temperature on a rotary evaporator for a total of 15 hours. The next day, the slurry was filtered through a 18.5 cm filter paper disk and the product was washed with 7 x 100 mL of anhydrous ethanol and finally dried under vacuum to constant weight. That's how it got

Is 85.8 g of white crystalline product, tlc _Rf 0.62 (methanol: water: acetic acid = 10: 1: 1) to give the title compound.

d) [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2 - {[(benzyloxy) carbonyl] amino} butyryl] amino} -6-hydroxy -hexánsavmetil ester

(S) -2-Amino-6-hydroxyhexanoic acid (1.0 g, 6.8 mmol) was suspended in methanol (20 mL) under argon, and chlorotrimethylsilane (1.9 mL, 15 mmol) was added with stirring at room temperature. suspension. The resulting solution was allowed to stir at room temperature for 18 hours, then concentrated in vacuo to a volume of about 3.5 mL. After adding 5 ml of acetonitrile, the solution was cooled to -10 ° C, followed by the addition of 15, Ν-diisopropylethylamine (9.15 ml, 23.8 mmol) and cooled to -40 ° C. This solution containing the methyl (S) 2-amino-6-hydroxyhexanoic acid is discarded.

In another flask, 2.117 g (6.8 mmol), all. To a solution of S-acetyl-N- (benzyloxycarbonyl) -homocysteine prepared as described in Example 1 (e) in 5 ml of acetonitrile and cooled to 0 ° C was treated with 1.20 ml (6.8 mmol) of Ν, Ν-diisopropyl. ethyl amine is added.

In a third flask, a solution of 0.104 g (0.68 mmol) of 1-hydroxybenzotriazole water (1/1) and 1.450 g (6.8 mmol) of [(methylsulfonyl) oxy] benzotriazole in acetonitrile cool to -18 ° C, then add dropwise a solution of the previously prepared (S) -acetyl-N - [(benzyloxy) carbonyl] homocysteine so that the internal temperature remains below -10 ° C throughout.

After the addition, the resulting solution was stirred at -18 ° C to -12 ° C for 3 hours, and then the (S) 2-amino-6-hydroxyhexanoic acid methyl ester solution and the mixture was allowed to warm slowly to 16 ° C over 18 hours. Thereafter, 50 ml of ethyl acetate and 50 ml

The mixture was poured into a mixture of hydrochloric acid (M), the immiscible phases were separated in a separatory funnel and the aqueous portion was extracted with ethyl acetate (50 ml). The combined organic solvent extracts were washed with 100 ml of 1M hydrochloric acid, 100 ml of saturated sodium bicarbonate solution and 100 ml of saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. To the resulting white solid was added 20 ml of tert-butyl methyl ether, the resulting slurry was stirred at room temperature for 4 hours, then filtered, washed with (tert-butyl) methyl ether and finally dried. 2.087 g of the title compound are obtained, m.p. 89-90 ° C.

e) [S- (R *, R *)] - 2 - {[4- (Acetylthio) -2 - {[(benzyloxy) carbonyl] amino} butyryl] amino} -5-formyl -valeriánsavmetil ester

Oxalyl dichloride (546 µL, 6.27 mmol) was dissolved in dichloromethane (16 mL), then 905 µL (12.54 mmol) of dimethyl sulfoxide were added dropwise at -65 ° C (internal temperature) over a period of about 12 minutes. of methylene chloride. The internal temperature is maintained at -65 ° C to -60 ° C throughout the addition. A solution of 1.90 g (4.18 mmol) of the compound prepared as in (d) above in dichloromethane (7 ml) is added over a period of about 20 minutes. The hydroxy compound was rinsed with additional 1 mL of dichloromethane and stirred at -65 ° C for 40 minutes, whereupon 3.7 mL (20.80 mmol) of N, N-diisopropylethylamine, a clear crude solution, was added. leads. After stirring for an additional 30 minutes at -65 ° C, the mixture was allowed to warm to -18 ° C over 2 hours, then quenched with 10% aqueous potassium bisulfate (30 ml) and finally at room temperature, 25 ml. After addition of water, the immiscible phases are separated. The aqueous layer was extracted with dichloromethane (2 x 25 mL), then the combined organic solvent extracts were washed with 10% aqueous potassium bisulfate (25 mL), saturated aqueous sodium bicarbonate (2 x 25 mL) and brine (25 mL). washed with brine, dried over magnesium sulfate, filtered and concentrated in vacuo. 1.84 g of a white solid are obtained.

f) [4S- (4a, 7a, 10a ')] - 4 - {[(Benzyloxy) carbonyl] amino} -5-oxooctahydro-7H-pyrido [2,1b] [1,3 ] thiazepine-7-carboxylic acid methyl ester

A dried flask, purged with argon, was charged with 1.76 g (3.89 mmol) of the compound prepared as in (e) above and 17 mL of methanol. The solution is cooled to 0 ° C and argon gas is bubbled through for 25 minutes and then 983 µl (4.28 mmol) of 25% w / w in methanol is added over about 15 seconds.

EN 217 960 Β of sodium methylate solution. After 1 hour, the reaction mixture was quenched with 1M hydrochloric acid (20 mL), shaken with ethyl acetate (35 mL), the two phases were separated and the aqueous portion was extracted with additional ethyl acetate (2 x 15 mL). The combined organic solvent extracts were washed with 15 mL of 1M hydrochloric acid and brine, dried over magnesium sulfate, filtered and the solvent was evaporated in vacuo. The resulting white foam (1.69 g) is [S- (R *, R *)] - 2 - {[2 - {[(benzyloxy) carbonyl] amino} -9-mercaptobutyryl] amino. } -5-oxohexanoic acid methyl ester.

The above foam, together with trifluoroacetic acid (305 μΐ, 3.95 mmol), was dissolved in dichloromethane (17 mL) and refluxed for 2.25 hours. After the reaction time, the mixture was cooled to room temperature, evaporated and the residue was dissolved in ethyl acetate (25 mL), washed with saturated sodium bicarbonate (2 x 20 mL) and brine, dried over magnesium sulfate, filtered and dried in vacuo. The remaining 1.50 g of a white foam is the title compound.

g) [4S- (4a, 7a, 10afi) -N-4-Amino-5-oxo-octahydro-7H-pyrido [2,1b] [1,3] thiazepine-7-carboxylic acid methyl ester

The above compound (f) was obtained in Example 8 (h) or all. Reaction with iodine trimethylsilane as described in Example 1 (h) removes the amino protecting group from the molecule. The product obtained is the desired amino compound.

Example 13 (S) -2-Phthalimido-6-hydroxyhexanoic acid

In addition to the procedure described in Example 1 (c), the compound may be prepared as follows.

730 g (2.2 mol) of 2-acetylamino-2- (4-acetoxybutyl) malonic acid diethyl ester prepared as in Example 12 (a) are dissolved in 300 ml of anhydrous ethanol and then added to 1600 g. ml of 6 M sodium hydroxide solution. The mixture is heated to 70-75 ° C for one hour and then most of the ethanol is distilled from the system at 90-95 ° C. The residue was cooled and about 1300 ml

It is acidified to pH 1.3 with hydrochloric acid M, then reheated to 90 ° C to 100 ° C, whereupon decarboxylation begins. Upon completion, the mixture was again cooled to room temperature.

To the above mixture was added 600 ml of 6M sodium hydroxide solution at 35 ° C, followed by 1M sodium hydroxide solution until the pH was exactly 7.5. To the approximately neutral solution (about 5300 ml), 0.6 g of porcine kidney acylase I was then added and stirring was continued at 35 ° C overnight, whereby the pH dropped to 7.25. The pH of the solution was restored to 7.5, an additional 300 mg of acylase was added and stirring was continued overnight. Thus, the next day, we find that the hydrolysis was about 90% complete. After adding 20 g of charcoal and 20 Celite, the mixture is heated to 85 ° C for 10 minutes, then cooled to 50 ° C and filtered. The filtrate (approximately 4900 ml) was cooled to 5 ° C, adjusted to pH 9.3 with solid sodium carbonate, and then added in one portion (263.04 g, 1.2 mol).

The pH is maintained at 9.3 by adding N-ethoxycarbonylphthalimide and, if necessary, sodium carbonate. After 2 hours at 5 [deg.] C., the reaction mixture is allowed to react at room temperature for an additional 3 hours, at which time the pH is reduced to 8.5 and the majority of the reagent is dissolved. The reaction mixture was filtered, cooled to 5 ° C and then acidified to pH 2.3 with 6M hydrochloric acid. The precipitate was filtered off, washed with cold water (200 mL) and dried in vacuo to give 220 g of the title compound.

Claims (5)

Compounds of formula (III): wherein: X is O or S; n is 1 or 2; m is 0 or 1; Y is methylene or oxygen, with the proviso that when Y is oxygen, m is 1; and R ₃ is hydrogen or C 1-7 alkyl; with the proviso that when X is O, n is 1 and Y is methylene then m is 1 and their salts. 2. A process for the preparation of a compound of formula III: wherein X is O or S; n is 1 or 2; m is 0 or 1; Y is methylene or oxygen, with the proviso that when Y is oxygen, m is 1; and R ₃ is a carboxy protecting group; with the proviso that when X is oxygen, n is 1 and Y is methylene, m is 1 and their salts are characterized in that (i) an amino acid of formula (XXII) and an amino acid ester of formula (XXIII): Pi represents an amino protecting group or, together with the nitrogen atom, represents an amine protecting group; P ₂ is a hydroxy or mercapto protecting group; and R 1 is a carboxy protecting group linked to a dipeptide of formula XXIV; ii) selectively removing the P ₂ protecting group from the molecule obtained in step i) and, if desired, converting the hydroxy compound into a mercapto compound; iii) the product of step ii) into a compound of formula (XLVilla) X, Y and R ₃ are in the scope, P _t is as defined above; and Iv) deprotecting the molecule obtained in step ifi); and optionally converting the resulting compound of formula (III) into a salt. A process according to claim 2 for the preparation of a compound of formula (III) wherein: X is sulfur; n is 1 or 2; Y is methylene; and m is 0 or 1; characterized in that (i) an amino acid of formula (XXIIa) with an amino acid ester of formula (XXIII): P 1 is an amino protecting group or, together with the nitrogen atom, represents an amine protecting group; and P ₂ is a hydroxy protecting group; R ₃ is a carboxy protecting group; and Y is a methylene group linked to a dipeptide of formula XXIVa; ii) selectively removing the P ₂ protecting group from the molecule obtained in step i); ifi) converting the hydroxy compound obtained in step ii) to a mercapto compound of the formula XXIVb; iv) cyclizing the mercapto compound prepared in step ifi) to a compound of formula XXVa; and v) deprotecting the compound obtained in step iv). A process according to claim 2 for the preparation of a compound of formula (III) wherein: X is sulfur; n is 2; Y is O; and m is 1; characterized in that i) an amino acid of formula XXIIa with an amino acid ester of formula XLVI in which P 1 is an amino protecting group or together with the nitrogen atom a protecting group which can be converted to an amine; P ₂ is a hydroxy protecting group; R ₃ is a carboxy protecting group; and Y is an oxygen atom to a dipeptide of formula (XLVII) X is O; P 1, P ₂ , Y, R ₃ and n are as defined above; ii) selectively removing the P ₂ protecting group from the compound obtained in step i); ifi) converting the hydroxy compound obtained in step ii) to a mercapto compound of formula XLVIIa; iv) cyclizing the mercapto compound obtained in step fii) to a compound of formula XLVIIb; and v) deprotecting the compound obtained in step iv). A process for the preparation of a compound of the formula (III) and salts thereof Y is methylene; X is O or S; n is 1 or 2; m is 0 or 1; and R ₃ is a carboxy protecting group; with the proviso that when X is oxygen and n is 1, then m is 1, characterized in that i) an amino acid of the formula XXII with a hydroxy amino acid ester of the formula XXVII - in the formulas Pi represents an amino protecting group or, together with the nitrogen atom, represents an amine protecting group; P ₂ is a hydroxy or mercapto protecting group; and n, m and R ₃ are linked to a dipeptide of formula XXVIII as defined herein; ii) converting the product of step i) to an aldehyde of formula XXIX by oxidation of the hydroxy group; ifi) selectively removing the P ₂ protecting group from the aldehyde molecule obtained in step ii); iv) cyclizing the product of step ifi) to a compound of formula XXV; and product of v) in step iv) cleaving the P _(a protecting group, and if desired the resulting compound of formula (III) into a salt.