Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
PROCESS FOR HCV PROTEASE INHIBITOR INTERMEDIATE
Document Type and Number:
WIPO Patent Application WO/2008/128921
Kind Code:
A1
Abstract:
The present invention relates to a new process for the preparation of diene compounds of the formula (I) wherein R1 is an amino protecting group and X is a halogen atom which may serve as intermediates for the manufacture of macrocyclic HCV protease inhibitors.

Inventors:
GANTZ, François (6 rue des Bosquets, Rixheim, Rixheim, F-68170, FR)
STAHR, Helmut (Friedrich-Neff-Strasse 3, Loerrach, 79539, DE)
Application Number:
EP2008/054518
Publication Date:
October 30, 2008
Filing Date:
April 15, 2008
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
F. HOFFMANN-LA ROCHE AG (Grenzacherstrasse 124, Basel, CH-4070, CH)
GANTZ, François (6 rue des Bosquets, Rixheim, Rixheim, F-68170, FR)
STAHR, Helmut (Friedrich-Neff-Strasse 3, Loerrach, 79539, DE)
International Classes:
C07D403/12; C07D487/04
Domestic Patent References:
WO2005037214A2
WO2008005511A2
Other References:
None
Attorney, Agent or Firm:
RAUBER, Beat (Grenzacherstrasse 124, Basel, CH-4070, CH)
Download PDF:
Claims:

Claims

1. Process for the preparation of a diene compound of the formula

wherein R 1 is an amino protecting group and X is a halogen atom comprising coupling the carbamate of formula

wherein X is a halogen atom with the carboxylic acid of the formula

or with a salt thereof to form the diene compound of the formula I. 2. Process of claim 1, characterized in that X is fluorine.

3. Process for the preparation of a diene compound of the formula

wherein R 1 is an amino protecting group and X is a halogen atom comprising the steps of a) reacting a dipeptide of the formula

wherein R 2 is an amino protecting group with carbonyl diimidazole and a compound of formula

wherein X is as above or a salt thereof to form an N-protected carbamate of the formula

wherein R 2 is an amino protecting group and X is as above; b) deprotecting the N-protected carbamate to form the carbamate of the formula

wherein X is as above; and c) coupling the carbamate of formula V with the carboxylic acid of the formula

or with a salt thereof to form the diene compound of the formula I.

4. Process of claim 3, characterized in that X is fluoro and R 1 and R 2 stand for a Boc- group.

5. Process of anyone of claims 1 to 4, characterized in that the diene compound of formula I has the structure

6. Process of anyone of claims 3 to 5, characterized in that the reaction in step a) is performed in the presence of a tertiary amine and an organic solvent.

7. Process of anyone of claims 3 to 6, characterized in that the deprotection of the N- protected carbamate of the formula IV in step b) is performed with an acid in an organic solvent.

8. Process of anyone of claims 1 to 6, characterized in that the coupling of the carbamate of formula V with the carboxylic acid of the formula VI is performed with a coupling agent selected from alkyl chloroformates, carboxylic acid halogenides or carbodiimides. 9. Process of claim 8, characterized in that the coupling agent is selected from isobutyl chloroformate or from pivaloyl chloride.

10. N-protected carbamate of the formula

wherein R 2 is an amino protecting group and X is a halogen atom.

11. Carbamate of the formula

wherein X is a halogen atom.

12. Use of the process of claims 1 to 9 in the preparation of the macrocyclic compound of formula

wherein X is a halogen atom.

13. Process for the preparation of the macrocyclic compound of formula

wherein X is a halogen atom comprising a process as claimed in the claims 1 to 9.

14. The invention as hereinbefore described.

Description:

PROCESS FOR HCV PROTEASE INHIBITOR INTERMEDIATE

The present invention relates to a new process for the preparation of diene compounds of the formula

wherein R 1 is an amino protecting group and X is a halogen atom which may serve as intermediates for the manufacture of macrocyclic HCV protease inhibitors of the formula.

wherein X is a halogen atom, particularly for the compound of the formula

I

which has been nominated for preclinical development.

The processes so far known for the preparation of the diene compound of the formula I suffers from instable intermediates, lack of purification steps, the use of corrosive reagents and intermediates, and hazardous and expensive coupling reagents (PCT

Publication WO 2005/037214). Therefore the known process is not suitable for large scale production.

Object of the present invention therefore was to find an improved process which is applicable on technical scale and delivers the diene of formula I in high quality and yield. It was found that this object could be reached with the process of the present invention as outlined below.

The following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the invention herein.

The term "amino protecting group" refers to any substituents conventionally used to hinder the reactivity of the amino group. Suitable amino protecting groups are described in Green T., "Protective Groups in Organic Synthesis", Chapter 7, John Wiley and Sons, Inc., 1991, 309-385. Suitable amino protecting groups are Fmoc, Cbz, Moz, Boc, Troc, Teoc or Voc. Preferred amino protecting group, as defined in R 1 and R 2 , is Boc.

The term "halogen" refers to fluorine, chlorine, bromine and iodine. The preferred definition for X is fluorine.

In a preferred embodiment the moiety of the formula

stands for

The process for the preparation of a diene compound of the formula

wherein R 1 is an amino protecting group and X is a halogen atom comprises coupling the carbamate of formula

wherein X is a halogen atom with the carboxylic acid of the formula

- A -

or with a salt thereof to form the diene compound of the formula I.

In a further embodiment of the invention the process for the preparation of a diene compound of the formula

wherein R 1 is an amino protecting group and X is a halogen atom comprises the steps a) reacting a dipeptide of the formula

wherein R 2 is an amino protecting group with carbonyl diimazole and a compound of formula

wherein X is as above or a salt thereof to form an N-protected carbamate of the formula

wherein R 2 is an amino protecting group and X is as above; b) deprotecting the N-protected carbamate of the formula IV to form the carbamate of the formula

wherein X is as above; and c) coupling the carbamate of formula V with the carboxylic acid of the formula

or with a salt thereof to form the diene compound of the formula I. In line with the structure of the preclinical candidate compound of formula VIII the diene compound of formula I has the preferred structure of the formula

The dipeptide of formula

which is the starting compound of the process of the present invention can be obtained by applying procedures known in the art. A possible approach is outlined in the scheme below:

3

For example the vinylcyclopropylcarboxylate i is treated with a suitable acid like sulfuric acid to form 2_and then coupled with 3 and isobutylchloroformate or pivaloylchloride in the presence of a tertiary amine like N-methylmorpholine to the dipepetide 4. The same reaction sequence, however with slightly different reaction conditions is known from the PCT Publication WO 2005/037214.

Step a)

Step a) requires the reaction of the dipeptide of the formula II with carbonyl diimidazole and with a compound of formula III to form the N-protected carbamate of formula IV. As mentioned above R 2 in the dipeptide of formula II preferably is Boc.

The compound of formula III can be applied as free base or as a salt formed with a suitable inorganic or organic acid.

Preferably a hydrohalogenide, more preferably a hydrochloride of the compound of formula III is used. Most preferred compound of formula III is the 4-fluoroisoindoline hydrochloride.

As a rule the reaction is performed in the presence of an organic solvent. Suitable organic solvents can be selected from polar aprotic solvents and aliphatic or aromatic hydrocarbons. The aromatic hydrocarbon toluene is the preferred solvent.

Step a) comprises in a first sequence the carbonyl diimidazole addition and in a second reaction sequence the carbamate formation. For the carbamate formation usually a tertiary amine is present. Suitable tertiary amines are trialkylamines such as triethylamine or diisopropylethylamine, whereby triethylamine is preferred.

The reaction is usually performed at a temperature in the range of -40 0 C to 130 0 C. Usually lower temperatures in the range of 0 0 C to 30 0 C are required for the carbonyl diimidazole addition and slightly higher temperatures in the range of 20 0 C to 80 0 C are applied for the carbamate formation.

The N-protected carbamate of formula IV can be isolated from the reaction mixture by applying methods known to the skilled in the art such as by extraction with a suitable organic solvent.

The N-protected carbamates of formula IV are compounds not known in the prior art and therefore are a further embodiment of the present invention.

In a preferred carbamate of formula IV X is fluoro and R 2 is Boc. Further preferred is the N-protected carbamate of the formula

Step b) Step b) requires deprotecting the N-protected carbamate of the formula IV to form the carbamate of formula V.

Deprotection can be achieved with an acid in an organic solvent.

Suitable acids can be selected from organic acids such as methane- or p-toluene sulfonic acid or inorganic acids such as hydrochloric-, hydrobromic- or sulfuric acid. Concentrated sulfuric acid is the preferred acid. Suitable organic solvents can be selected from polar protic or aprotic solvents or from aliphatic or aromatic hydrocarbons or mixtures thereof. Ethylacetate was found to be the preferred solvent.

The reaction is usually performed at a temperature in the range of 0 0 C to 80 0 C.

The carbamate of formula V can be isolated from the reaction mixture by applying methods known to the skilled in the art such as by extraction with a suitable organic solvent.

Further purification of the product can be obtained by crystallization of the product in a suitable solvent such as in polar protic or aprotic solvents or from aliphatic or aromatic hydrocarbons or mixtures thereof. Toluene was found to be the preferred solvent. The carbamates of formula V are compounds not known in the prior art and therefore are a further embodiment of the present invention.

In a preferred carbamate of formula V X is fluoro. Further preferred is the carbamate of the formula

Step c)

Step c) requires coupling of the carbamate of formula V with the carboxylic acid of the formula

or with a salt thereof to form the diene compound of the formula I.

The carboxylic acid of formula VI or the salts thereof are as a rule commercially available. Suitable salts of the carboxylic acid of formula VI are ammonium salts formed with secondary or tertiary amines such as with dicyclohexylamine or N-methylmorpholine.

The coupling reaction as a rule is performed according to methods known in the art for peptide synthesis (e.g. Houben Weyl "Synthesis of Peptides and Peptidomimetics", Chapter 3, Thieme Stuttgart New York, 2004, 425-588) The coupling reaction requires a coupling agent which usually is selected from alkyl chloroformates such as methyl-, ethyl-, isopropyl-, sec-butyl, isobutyl- and cyclopentylchloroformate, from carboxylic acid halogenides such as pivaloylchloride, from carbodiimides such as dicyclohexylcarbodiimide, diisopropylcarbodiimide and N- [3- (dimethylamino)propyl]-iV-ethylcarbodiimide hydrochloride (EDC). Also N- [(1H- benzotriazol-1-yl) (dimethylamino) methylene] -N-methylmethanaminium tetrafiuoroborate N-oxide (TBTU) or 2-chloro-4,6-dimethoxy-l,3,5-triazine (CDMT) can be used as coupling agent.

The carbodiimides may be used together with additives such as with 1,2,3- benzotriazol-1-ol (HOBt) or N-hydroxysuccinimide. Preferred coupling agents are isobutylchloroformate or pivaloylchloride.

A tertiary amine such as N-methylmorpholine, triethylamine or diethlymethylamine, dimethylethylamine, trimethylamine, dimethylpropylamine, diisopropylmethylamine, N,N-dimethylpiperazine, N-methylpiperidine, preferably N-methylmorpholine or triethylamine may be present. In a preferred embodiment the reaction can be performed without the addition of a tertiary amine.

The reaction is performed in suitable organic solvent which can be selected from polar aprotic solvents or from aliphatic or aromatic hydrocarbons or mixtures thereof such as ethylacetate, tetrahydrofuran, dimethylformamide, methylenechloride, toluene, preferably from ethylacetate and tetrahydrofuran or mixtures thereof. The reaction is usually performed at a temperature in the range of -40 0 C to 130 0 C, preferably -40 0 C to 60 0 C.

The diene of formula I can be isolated from the reaction mixture by applying methods known to the skilled in the art such as by extraction with a suitable organic solvent, e.g. with toluene. In a further embodiment the process of the present invention can be used in the preparation of the macrocyclic compound of formula

wherein X is a halogen atom, preferably for the preparation of the macrocyclic compound of formula

In a further embodiment the invention comprises a process for the preparation of the macrocyclic compound of formula

wherein X is a halogen atom which comprises the process of the present invention.

A suitable way to convert the diene of formula I into the macrocyclic compounds of formula VII or VIII is disclosed in the PCT Publication WO 2005/037214.

The following examples shall illustrate the invention without limiting it.

Examples

Example 1

(lR,25)-l-Amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester A suspension of 5.11 g (20.0 mmol) of (l_R,2S)-l-tert-butoxycarbonylamino-2-vinyl- cyclopropanecarboxylic acid ethyl ester (commercially available from Synthetech Oregon, USA) in 1.94 mL of ethyl acetate was cooled to 8 0 C using an ice bath. Then a solution of 2.17 g (21.0 mmol) of sulfuric acid in 4.0 mL of ethyl acetate was added in 5 min. The ice bath was removed and the reaction mixture was stirred for 30 min at room temperature and 1 h at 50 0 C. The reaction mixture was then cooled to room temperature and used in the next step without further purification.

Example 2

(25,4R) -2- ( ( 1_R,25) - 1 -Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4-hydroxy- pyrrolidine-1-carboxylic acid tert-butyl ester

To a solution of 4.87 g (21.0 mmol) of Boc-(2S,4_R)-hydroxyproline in 25 mL of THF 2.03 mL (20.0 mmol) of N-methylmorpholine was added. A suspension was formed. The mixture was cooled to -23 0 C and 2.85 g (20.0 mmol) of isobutylchloroformiate was added. After stirring for 10 min additional 4.25 g (42.0 mmol) of N-methylmorpholine was added. To this mixture the solution of ( l_R,2S)-l-Amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester prepared in example 1 was added at a temperature of -15 0 C within 5 min. The reaction mixture was stirred for 2.5 h at 0 0 C. Salts were filtered off and the filtrate was treated with 20 mL of aqueous HCl (0.5 N). The solvents were removed at 50 0 C under reduced pressure using a rotary evaporator and the residue was extracted twice with 50 mL of ethyl acetate. The extract was washed with 40 mL of water and 40 mL of aqueous sodium carbonate solution ( 10 wt %), and dried over sodium sulfate. Finally the solvent was removed completely to give 8.19 g of (2S,4_R)-2-(( l_R,2S)-l-ethoxycarbonyl-2-vinyl- cyclopropylcarbamoyl)-4-hydroxy- pyrrolidine- 1-carboxylic acid tert-butyl ester as a yellow oil. The product was used in the next step without further purification.

Example 3

4-Fluoro-l,3-dihydro-isoindole-2-carboxylic acid f3#,5S)-l-ferf-butoxycarbonyl-5- ( ( IR,2S) - 1 -ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin-3-yl ester

8.19 g of crude (2S,4i?)-2-(( li?,2S)-l-ethoxycarbonyl-2-vinyl- cyclopropylcarbamoyl)-4-hydroxy- pyrrolidine- 1-carboxylic acid tert-butyl ester was dissolved in 60 mL of toluene and 4.25 g carbonyldiimidazole (26.2 mmol) was added in portions at a temperature of 22-25 0 C. The reaction mixture was stirred for 1.5 h at ambient temperature. Then 3.66 g (21.0 mmol) of 4-fluoroisoindoline hydrochloride was added in portions followed by 3.1 mL of triethylamine. The resulting suspension was heated to 52 0 C bath temperature. After stirring for 3 h at this temperature the reaction mixture was cooled with an ice bath. 70 mL of aqueous HCl ( IM) were added. The mixture was extracted with 50 mL of toluene. The separated aqueous layer was extracted twice with 50 mL toluene. The combined toluene extracts were washed with 30 mL of water and 30 mL of an aqueous solution of sodium carbonate (5 wt %). The toluene extract was dried with sodium sulfate, filtered, and the solvent was completely removed. 9.21 g of 4-fluoro- l,3-dihydro-isoindole-2-carboxylic acid (3R,5S)-l-tert-butoxγcarbonγ\-5-(( lR,2S)-l- ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin-3-yl ester was obtained as a grey solid, which was used in the next step without further purification.

MS: 532.3 (M + + H). 1 H-NMR (400 MHz, DMSO-D6, 79.2 0 C): 8.40 (s, IH), 7.37-7.31 (m, IH), 7.16 (d, J=7.5 Hz, IH), 7.09-7.05 (m, IH), 5.73-5.64 (m, IH), 5.24 (dd, J=17.2, 1.6Hz, IH), 5.18 (m, IH), 5.08 (dd, J=IOA, 1.6Hz, IH), 4.67 (m, 4H), 4.22 (t, J=7.7Hz, IH), 4.11- 4.00 (m, 2H), 3.66 (dd, J=I 1.9, 4.7Hz, IH), 3.54 (d,br, 12.1Hz, IH), 2.37-2.28 (m, IH), 2.19-2.11 (m, 2H), 1.63 (dd, J=7.95, 5.25Hz, IH), 1.38 (s, 9H), 1.28 (dd, J=9.4, 5.1Hz, IH), 1.16 (t, J=7.0Hz, 3H).

Example 4

4-Fluoro-13-dihvdro-isoindole-2-carboxylic acid (3R,5S)-5-(( lR,2S)-l- ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-pyrrolidin-3-yl ester

A solution of 2.15 g (21.0 mmol) of sulfuric acid in 3.9 mL of ethyl acetate was added to a suspension of 9.21 g of crude 4-fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3R, 5S) - 1 -terf-butoxycarbonyl-5- ( ( IR,2S) - 1 -ethoxycarbonyl-2-vinyl-cyclopropyl- carbamoyl)-pyrrolidin-3-yl ester in 31 mL of ethyl acetate, which was cooled with an ice bath. The ice bath was removed and the reaction mixture was heated to 50 0 C for 3 h until all starting material had been consumed. To the reaction mixture an aqueous solution of sodium carbonate ( 10 wt %) was added. Phases were separated and the aqueous layer was extracted three times with ethyl acetate. The combined organic extracts were evaporated to dryness and the residue was dissolved in 85 mL of toluene and heated to 102 0 C. The solution was slowly cooled to 2 0 C. Crystallization started at 53 0 C. The crystals were filtered off and dried under reduced pressure to yield 6.62 g (77% over four steps starting from 20.0 mmol ( li?,2S)-l-tert-Butoxycarbonylamino-2-vinyl-cyclopropanecarbo xylic acid ethyl ester; yield not corrected for assay; assay: 97.9% area HPLC) of 4-fluoro-l,3-dihydro- isoindole-2-carboxylic acid (3i?,5S)-5-(( li?,2S)-l-ethoxycarbonyl-2-vinyl-cyclopropyl- carbamoyl)-pyrrolidin-3-yl ester as grey crystals.

MS: 432.2 (M + + H). 1 H-NMR (400 MHz, CDCl 3 ): 8.17 (d, J=4.0 Hz, IH), 7.31-7.25

(m, IH), 7.08-6.95 (m, 2H), 5.82-5.73 (m, IH), 5.31 (dd, J=16.4, 1.2 Hz, IH), 5.29 (m, IH), 5.13 (dd, J=IOJ, 1.7Hz), 4.82-4.65 (m, 2H), 4.23-4.07 (m, 2H), 3.98 (m, IH), 3.28 (d, 13.0 Hz, IH), 3.07-3.02 (m, IH), 2.46-2.40 (m, IH), 2.30 (s, br, IH), 2.26-2.17 (m, IH), 2.12 (m, IH), 1.92 (dd, J=7.9, 5.5 Hz, IH), 1.6-1.56 (m, IH), 1.24 (t, J=7.1Hz, 3H).

Example 5

4-Fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3R,5S)-l-((S)-2-tert- butoxycarbonylamino-non-8-enoyl)-5-(( l-R,2S)-l-ethoxycarbonyl-2-vinyl- cyclopropylcarbamoyl) -pyrrolidin-3-yl ester

A solution of 1.15 g (2.55 mmol) (S)-2-tert-butoxycarbonylamino-non-8-enoic acid dicyclohexylammonium salt (commercially available from Synthetech Oregon, USA) and 469 mg (4.64 mmol) N-methylmorpholine in 9.0 mL of THF was added dropwise to a solution of 302 mg (2.53 mmol) pivaloyl chloride in 1.5 mL of THF maintaining the temperature at 20-25 0 C. The suspension was stirred for 45 min, then cooled to 0 0 C. A solution of 1.00 g (2.32 mmol) of 4-fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3-R,5S)-5-((l-R,2S)-l-ethoxycarbonyl-2-vinyl-cyclopropylcar bamoyl)-pyrrolidin-3-yl ester in 13 mL of THF was added to the mixed anhydride at 0 0 C within 25 min. The mixture was first stirred for 2.5 h at 2 0 C, then for 19 h at 26 0 C. 9.5 mL water and 14.8 mL aqueous HCl (0.5 N) were added. The phases were separated and the aqueous layer was extracted with toluene (3 x 3 mL). The combined organic layers were washed with 2 mL of water, 5 mL of aqueous sodium carbonate (5 wt %) and dried over sodium sulfate. The solvent was removed at 50 0 C under reduced pressure using a rotary evaporator. The resulting oil was finally dried under oil pump vacuum yielding 1.75 g (88.3%) of 4-fluoro-l,3-dihydro- isoindole-2-carboxylic acid (3J?,5S)-l-((S)-2-tert- butoxycarbonylamino-non-8-enoyl)-5- ((l-R,2S)-l-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-pyr rolidin-3-yl ester as a brown resin with an assay of 80.5 wt %.

Example 6

(lR,25)-l-Amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester A solution of 153.2 g (600 mmol) of (l_R,2S)-l-tert-butoxycarbonylamino-2-vinyl- cyclopropanecarboxylic acid ethyl ester (commercially available from Synthetech Oregon, USA) in 80 mL of ethyl acetate was dosed to a solution of 65.1 g (630 mmol) of sulfuric acid in 480 mL of ethyl acetate at 20-25 0 C. The reaction mixture was stirred for 60 min at 42-48 0 C. After complete conversion the reaction mixture was cooled to 18-22 0 C. Then 127.5 g (1260 mmol) of triethylamine was dosed at 20-30 0 C. Finally 100 mL of dimethylformamide was added. The reaction mixture was used in the next step immediately after preparation.

Example 7

(25,4,R) -2- ( ( IR,2S) - 1 -Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4-hydroxy- pyrrolidine- 1-carboxylic acid tert-butyl ester

To a suspension of 138.8 g (600 mmol) of Boc-(2S,4_R)-hydroxyproline in 890 mL of ethyl acetate 63.7 g (630 mmol) of N-methylmorpholine was added at 20-30 0 C. To this mixture 72.3 g (600 mmol) of pivaloyl chloride was added at 15-22 0 C within 10-20 min followed by 50 mL of ethyl acetate. The mixture was stirred at 20-25 0 C for 60-75 min. Then the emulsion of ( IR,2S) - 1 -amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester

prepared in example 6 was added at a temperature of 20-30 0 C within 5-30 min. The dosing funnel was rinsed with 20 mL dimethylformamide and 100 mL ethyl acetate. The reaction mixture was stirred for 4 h at 20-25 0 C. After complete conversion 450 mL of aqueous HCl (IN) was added. Phases were separated and the aqueous layer was extracted with 600 mL of ethyl acetate. The organic phase was washed with 200 mL of water, 400 mL of aqueous sodium hydroxide (2M) and 200 mL of water. The combined organic phases were concentrated to 400 mL under reduced pressure. To the residue 1500 mL of toluene was added and the mixture was concentrated to 1000 mL under reduced pressure. To the residue 720 mL of toluene was added to yield the (2S,4_R)-2-((l_R,2S)-l-ethoxycarbonyl-2- vinyl-cyclopropylcarbamoyl)-4-hydroxy- pyrrolidine- 1-carboxylic acid tert-butyl ester as a 12-13 wt % solution in toluene.

Example 8

4-Fluoro-l,3-dihydro-isoindole-2-carboxylic acid f3R,55)-l-fe?t-butoxycarbonyl-5- ( ( IR,2S) - 1 -ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin-3-yl ester To a solution of (2S,4_R)-2-((l_R,2S)-l-ethoxycarbonyl-2-vinyl-cyclopropylcar ba- moyl)-4-hydroxy- pyrrolidine- 1-carboxylic acid tert-butyl ester in toluene (as obtained in example 7) 121.1 g carbonyldiimidazole (724 mmol, purity 97 wt %) was added in six portions at 20-25 0 C. After the addition of each portion the reaction mixture was stirred for 20 min. The funnel was rinsed with 100 mL of toluene and the reaction mixture was stirred at 20-25 0 C for 1 h. Then 112.5 g (648 mmol) of 4-fluoroisoindoline hydrochloride was added followed by 30.4 g (300 mmol) of triethylamine and 100 mL of toluene. The resulting suspension was heated to 48-52 0 C temperature and stirred at this temperature for 5 h. After the conversion is completed the reaction mixture was heated to 57-62 0 C and 546 g of aqueous HCl (2M) was added. The phases were separated and the organic layer was washed with 500 mL of water. The organic phase was then concentrated at a jacket temperature of 60 0 C under reduced pressure to a residual volume of 600 mL. To the residue 1140 mL of ethyl acetate was added. The obtained solution of 4-fluoro-l,3- dihydro-isoindole-2-carboxylic acid (3R, 5S) - 1 -terf-butoxycarbonyl-5- ( ( IR,2S) - 1 - ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin-3-yl ester was used in the next step without further purification.

Example 9

4-Fluoro-13-dihvdro-isoindole-2-carboxylic acid (3R,5S)-5-((lR,2S)-l- ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin-3-yl ester

A solution of 64.4 g (630 mmol) of sulfuric acid in 120 mL of ethyl acetate was added to a suspension of 1.7 L 4-fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3R,5S)-l-tert-

butoxycarbonyl-5-( ( IR,2S) - 1 -ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -pyrrolidin- 3-yl ester (theoretical amount: 600 mmol; as obtained from example 8) in ethyl acetate at 50-60 0 C. The dropping funnel was rinsed with 80 mL of ethyl acetate and the reaction mixture was stirred for 3 h at 60 0 C. To the reaction mixture 800 mL of an aqueous solution of sodium carbonate ( 10 wt %) was added. Phases were separated and the organic layer was washed with 400 mL of water. The organic layer was concentrated at a jacket temperature of 105 0 C to a residual volume of 450 mL. To the residue 1000 mL of toluene was added. To initiate crystallization the temperature was decreased to 60 0 C and maintained for 30 min at this value. The suspension is cooled to -2 0 C and stirred for 1 h at this temperature. The crystals were collected on a suction filter and washed three times with a total of 400 mL of toluene. After drying at 53 0 C under reduced pressure 198.2 g of 4-fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3J?,5S)-5-((lJ?,2S)-l- ethoxycarbonyl-2- vinyl-cyclopropylcarbamoyl)-pyrrolidin-3-yl ester were obtained as colorless crystals in a purity of 98.0 wt % and a yield of 75% over 4 steps. Example 10

4-Fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3-R,5S)-l-((S)-2-tert- butoxycarbonylamino-non-8-enoyl)-5-((l-R,2S)-l-ethoxycarbony l-2-vinyl- cyclopropylcarbamoyl) -pyrrolidin-3-yl ester

A solution of 57.7 g (128 mmol) (S)-2-tert-butoxycarbonylamino-non-8-enoic acid dicyclohexylammonium salt (commercially available from Synthetech Oregon, USA) in 350 mL of THF was added dropwise to a solution of 15.1 g (125 mmol) pivaloyl chloride in 200 mL of THF maintaining the temperature at 0-5 0 C. The dosing funnel was rinsed with 50 mL of THF and the suspension was stirred for 90 min at 5 0 C jacket temperature. The temperature was decreased to -5 0 C and 50.0 g (116 mmol) of 4-fluoro-l,3-dihydro- isoindole-2-carboxylic acid (3i?,5S)-5-((li?,2S)-l-ethoxycarbonyl-2-vinyl- cyclopropylcarbamoyl) -pyrrolidin-3-yl ester was added in five portions to the mixed anhydride at -5 0 C to 0 0 C within 30 min. The funnel was rinsed with 25 mL of THF. The suspension was first stirred for 30 min at 0 0 C, then for 6 h at 23 0 C. After complete conversion 150 mL of water was added. The reaction mixture was concentrated at 50 0 C jacket temperature under reduced pressure to a residual volume of 300 mL. After cooling to 23 0 C 500 mL of toluene and 58 mL of aqueous HCl (IM) were added. The suspension was filtered and the reaction vessel and filter cake were washed with 220 mL of toluene. The phases of the filtrate were separated and the aqueous layer was extracted with 150 mL of toluene. The organic layers were washed separately with 200 mL of aqueous sodium carbonate (10 wt %) and 58 mL of aqueous HCl ( IM). The combined organic layers were concentrated at 50 0 C under reduced pressure using a rotary evaporator to afford 143.3 g of crude (54.5 wt %) 4-fluoro-l,3-dihydro-isoindole-2-carboxylic acid (3J?,5S)-l-((S)-2-tert-

butoxycarbonylamino-non-8-enoyl) -5-( ( 1_R,2S) - 1 -ethoxycarbonyl-2-vinyl-cyclopropyl- carbamoyl)-pyrrolidin-3-yl ester in a yield of 98.4%.