WO2000018770A1 - Pyrrolopyrrolone derivatives as antiviral agents - Google Patents

Abstract

The present invention relates to therapeutically active bicyclic compounds, processes for the manufacture of said compounds, pharmaceutical formulations containing said compounds and the use of said compounds in the treatment and prophylaxis of viral infections, more particularly infections caused by viruses which encode for a serine protease enzyme, especially viruses of the Herpes family. The invention provides compound of general formula (I) (relative stereochemistry shown) and pharmaceutically acceptable derivatives thereof, wherein R represents H substituted or unsubstituted C1-3 alkyl; R1 represents optionally substituted heteroaryl or fused heteroaryl, with one to four heteroatoms, or R9CO; R3 represents C1-6alkyl, C1-6alkenyl, optionally substituted three to seven member, preferably four to seven member, N-heterocycle containing up to two heteroatoms, (a) or (b); and R4, R5, R6, R7 and R9 independently represent H or a substituted or unsubstituted alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heteroaryl group as defined in the specification.

Description

PYRR0L0PYRR0L0NE DERIVATIVES AS ANTIVIRAL AGENTS

The present invention relates to therapeutically active bicyclic compounds, processes for the manufacture of said compounds, pharmaceutical formulations containing said compounds and the use of said compounds in chemotherapy. In particular, we have found a novel group of bicyclic compounds which are effective in the treatment and prophylaxis of viral infections, more particularly infections caused by viruses which encode for a serine protease enzyme, especially viruses of the Herpes family.

The Herpes family of viruses is responsible for a wide range of infectious diseases in several species especially chicken pox, shingles, retinitis, pneumonitis and keratitis in humans and diseases of the skin and mucosa, including keratitis in rabbits, herpetic encephalitis in mice, Herpes viruses include HSV1 and HSV2 (Herpes Simplex Virus type 1 and type 2), hCMV (human cytomegalovirus), VZV (varicella zoster virus), EBV (Epstein-Barr virus) HHV6 and HHV8 (human herpes viruses, types 6 and 8).

All herpes viruses encode a serine protease which is crucial for viral replication as it cleaves the assembly protein precursor during capsid maturation. Protease deficient mutants do not cleave this scaffold protein thus giving rise to immature virions. We have found that inhibitors of this protease can have a similar effect thus preventing formation of mature, infectious viral progeny in infected cells.

We now present a novel group of bicyclic compounds which are inhibitors of herpes virus proteases and are thus of benefit in the treatment, prophylaxis and suppression of virus infections including genital herpes, cold sores, chicken pox, shingles, retinitis, pneumonitis and keratitis caused by viruses of the herpes family. The compounds show broad spectrum activity against herpes viruses including HSV 1 and 2, hCMV and VZV.

Thus, according to one aspect of this invention, we provide a compound of the general formula (I) (relative stereochemistry shown):

and pharmaceutically acceptable derivatives thereof, wherein:

R represents H or substituted or unsubstituted C^ alkyl;

R, represents optionally substituted heteroaryl or fused heteroaryl, with one to four heteroatoms, or R₉CO;

R₂ represents -S0₂R₃;

R₃ represents C^alkyl, C^alkenyl, optionally substituted three to seven member, preferably four to seven member, N-heterocycle containing up to two heteroatoms,

R₄

⁴\ ^R6\

N — or T

« R₇

R₄ and R₅ independently represent H or a substituted or unsubstituted group selected from C^alkyl optionally including one or more heteroatoms, C^ ₆hydroxyalkyl,

(optionally containing from 1 to 4 heteroatoms), C^alkylCOR₁₃ C^alkylCH₂OR₁₃, C^alkylNHCOR₁₃₎

₃alkylaryl, heteroaryl and C^alkylheteroaryl;

R₆ and R₇ independently represent H or a substituted or unsubstituted group selected from C^alkyl optionally including one or more heteroatoms, C^ ₆hydroxyalkyl,

R₉ represents H or a substituted or unsubstituted group selected from C^alkyl, C^alkenyl, C₃.₇cycloalkyl, C₆.₁₀fused cycloalkyl, C₆-₁₀fused cycloalkenyl, heteroaryl or fused heteroaryl containing one to four heteroatoms, and C,. ₃alkylaryl;

R₁₃ and R₁₄ independently represent H or a substituted or unsubstituted group selected from C_halky!, C₃.₇cycloalkyl, C^alkenyl, C.^haloalkyl, aryl, C^alkylaryl, heteroaryl or C^alkylheteroaryl;

R₁₅ and R₁₆ independently represent H or a substituted or unsubstituted C^alkyl which may, together with the nitrogen atom to which they are attached, form a ring optionally containing one further heteroatom.

In preferred embodiments, R is C^alkyl, particularly methyl.

Preferably, R^ is selected from

wherein R' independently represent H, C^alkyl or C^hydroxyalkyl and R" independently represent H, OH, -OCOC^alkyl, -OCONHC^alkyl, or may together form a double bond;

optionally substituted with one or more groups selected from halogen, hydroxy, C^alkyl, C^hydroxyalkyi, C^alkylNHCORn, C^alkylOCOR^, C,_. βalkylOCONHR,, C_{1 ϊ}alkylNHS0₂R₁₁, C^alkylNHCONHR^, wherein R„ represents H, C^alkyl or C^haloalkyl.

Particular R groups include:

Q = CH₂OH,

Q, = CH₂OH,

CH₂NHCO e, CH₂NHCOMe, CH₂CH₂OH, CH₃, CH₂CH₂NHCO e, CH₂OAc, CH₂NHS0₂CH2Ph, CH₂OCONHMe,

CH₂NHS0₂Me,

CH₂OCONHEt, CH₂NHCONHEt,

CH₂NHS0₂CF₃

CH₂NHCO

In certain preferred compounds, R₃ represents optionally substituted four to seven member N-heterocycle containing up to two heteroatoms. Particularly preferred R₃ groups include

Preferred R₄ and R₅ groups include optionally substituted C^alkyl, aryl and C^ ₆alkylCONR₁₃R₁₄ wherein R₁₃ and R₁₄ independently represent H or a substituted or unsubstituted group selected from C^alkyl, aryl, C_Malkylaryl and G,. ₄alkylheteroaryl, particularly benzyl, more particularly benzyl bearing one or more substituents (which may be the same or different) selected from halogen, trihalomethyl, methoxy and nitro.

Where R₄ or R₅ is alkyl, suitable alkyl substituents are e.g. methyl, ethyl, n- propyl, isopropyl, n-butyl, isobutyl. Where R₄ or R₅ is aryl, suitable substituents include cyano, C,.₃alkoxy,

wherein R₈ and R₁₀ independently represent H, C^alkyl, C_1-3alkenyl, C,.

₃haloalkyl, or together form a ring which may optionally include a further heteroatom.

In particularly preferred embodiments, R₄ represents methyl and R₅ is selected from heteroarylCH₂NHCOCH₂-, heteroarylNHCOCH₂-, arylCH₂NHCOCH₂-, 4- Me₂NCH₂C₆H₄CH₂-, or HOCH₂CH(OH)CH₂-.

Formula (I) above shows the relative stereochemistry of the chiral centres. Generally, we prefer to provide a compound of formula (I) in enantiomerically pure cis-trans form with SRS stereochemistry as shown below, in which the hydrogens at the two ring fusion carbons are trans to one another and the hydrogen at the R- substituted carbon is cis to that at the adjacent ring fusion carbon. The absolute configuration is set out below:

When used herein "alkyl" includes branched as well as straight chain saturated hydrocarbon groups.

When used herein "alkenyl" includes branched as well as straight chain hydrocarbon groups containing one or more carbon-carbon double bonds.

When used herein "halogen" includes F, CI, Br and I.

Where stated, nitrogen atoms may be substituted by C^alkyl or -CO-C^alkyl groups and carbon atoms may be substituted by halogen, cyano or hydroxy groups, C=0, -OC^haloalkyl, NH₂, NHR₈R₁₀, N0₂, C^alkyl, C^haloalkyl, C.. ₄alkoxy, C^hydroxyalkyl, SR₈, S0₂R₈, C(0)R₈, C0₂R₈, C^alkylC0₂R₈, C^ ₆alkylOCOR₈, C^alkylOCONHR₈, C_1-6alkylNHCONHR₈, C^alkylNHS0₂R₈, C,_ ₆alkylCONHR₈, C^alkylCONR₈R_10> C^alkylNR₈R₁₀, CONHR₈, C0₂NHR₈, and C,_ ₆alkylNHCOR₈ wherein R₈ and R₁₀ independently represent H, C_1-3alkyl, C,. ₃alkenyl, C^haloalkyl, or together form a ring which may optionally include a further heteroatom.

When used herein "aryl" includes aromatic groups having up to two rings, including phenyl and naphthyl, and arylalkyl, heteroaryl are to be read accordingly. Thus, heteroaryl includes aromatic groups having up to two rings containing one or more (e.g. 1-4) similar or dissimilar heteroatoms e.g. pyridine, thiadiazole, thiophene, benzoxazole and benzothiazole. When used herein "cycloalkyl" includes carbocyclic groups having up to two rings, and carbocycle, carbocyclic, alkylcycloalkyl, heterocyclic, heterocycle are to be read accordingly. Thus, heterocyclic includes cyclic groups having up to two rings containing one or more (e.g. 1-4) similar or dissimilar heteroatoms e.g. morpholine, piperidine, piperazine, pyrrolidine, azetidine, homopiperidine, homopiperazine.

By "a pharmaceutically acceptable derivative" is meant any pharmaceutically or pharmacologically acceptable salt, ester or salt of such ester of a compound according to the invention, or any compound which, upon administration to the recipient, is capable of providing (directly or indirectly) a compound according to the invention, or an antivirally active metabolite or residue thereof.

Preferred esters of the compounds according to the invention are independently selected from the following groups: (1) carboxylic acid esters in which the non- carbonyl moiety of the carboxylic acid portion of the ester grouping is selected from straight or branched chain alkyl (for example, n-propyl. t-butyl, or n-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (for example, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for example, phenyl optionally substituted by, for example, halogen, C^alkyl, or C^alkoxy or amino); (2) sulphonate esters, such as alkyl- or aralkylsulphonyl (for example, methanesulphonyl); (3) amino acid esters (for example, L-valyl or L-isoleucyl);

(4) phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate esters may be further esterified by, for example, a C^alcohol or reactive derivative thereof, or by a 2,3-di(C_6-24)acyl glycerol.

In such esters, unless otherwise specified, any alkyl moiety present advantageously contains from 1 to 18 carbon atoms, particularly form 1 to 6 carbon atoms, more particularly from 1 to 4 carbon atoms. Any cycloalkyl moiety present in such esters advantageously contains from 3 to 6 carbon atoms. Any aryl moiety present in such esters advantageously comprises a phenyl group. Preferred carboxylic acid esters according to the present invention include the acetate, butyrate and valerate esters. L-valyl is a particularly preferred amino acid ester.

Examples of suitable solvates of the compounds of formula (I) include hydrates and alcohol solvates such as methanolates or ethanolates.

Any reference to any of the above compounds also includes a reference to a pharmaceutically acceptable salt thereof.

Where compounds of formula (I) are able to form physiologically acceptable salts, these are included within the present invention. Suitable physiologically acceptable salts of the compounds of formula (I) include inorganic base salts such as alkali metal salts (for example sodium and potassium salts) and ammonium salts and organic base salts. Suitable organic base salts include amine salts such as trialkylamine (e.g. triethylamine), dialkylamine (e.g. dicyciohexylamine), optionally substituted benzylamine (e.g. phenylbenzylamine or p-bromobenzylamine), procaine, ethanolamine, diethanolamine, N- methylglucosamine and tri(hydroxymethyl)methylamine salts and amino acid salts (e.g. lysine and arginine salts). Suitable inorganic and organic acid salts include the hydrochloride, trifluoroacetate and tartrate.

It is to be understood that the present invention covers all combinations of particular and preferred groups described hereinabove.

Certain compounds of formula (I) embodying the invention have structures as follows:

(CH₃)₂CH, H

(CH₃)₂CHCH₂, CH₃

Ph, CH₃

MeCH₂, MeCH₂ m-NCC₆H₄CH₂, CH₃ HOCH₂CH(OH)CH₂, CH₃

R₄, R₅ = CH₃, CH₃ CH₃, H

R₁₇ = H, C₁_₃alkyl, C₁.₃alkenyi

Intermediates and processes described herein form further aspects of the invention.

Compounds of formula (I) are of potential therapeutic benefit in the treatment and amelioration of the symptoms of many herpes virus diseases. Such diseases particularly include chicken pox and shingles (varicella and herpes zoster viruses, respectively), keratitis in rabbits, herpetic encephalitis in mice, cutaneous herpes in guinea pigs, cold sores and genital herpes in humans (herpes simplex virus), retinitis, pneumonitis and keratitis in humans (hCMV), as well as diseases caused by Epstein Barr Virus (EBV), human herpes virus 6 (HHV 6), HHV 7 and HHV 8. Compounds of the invention may also be useful for the treatment or prophylaxis of multiple sclerosis, in which herpes viruses have been implicated, and cardiovascular system diseases which hCMV has been implicated, such as thrombosis, arteriosclerosis and particularly restenosis (recurrent narrowing or occlusion of a coronary valve or vessel).

As indicated above, compounds of formula (I) are useful in human or veterinary medicine, in particular as inhibitors of viral serine proteases, in the management of herpes family virus infections.

Thus, there is provided as a further aspect of the present invention a compound of formula (I) or a physiologically acceptable salt or solvate thereof for use in human or veterinary medicine, particularly in the treatment of conditions caused by viruses of the Herpes family, such as HSV, VZV or CMV infections.

It will be appreciated that references herein to treatment extend to prophylaxis, prevention of recurrence and suppression of symptoms as well as the treatment of established conditions.

According to another aspect of the invention, there is provided the use of a compound of formula (I) or a physiologically acceptable salt or solvate thereof in the manufacture of a medicament for the treatment of conditions caused by viral infections, more particularly caused by viruses of the Herpes family, such as HSV, VZV or CMV infections.

In a further or alternative aspect there is provided a method for the treatment of a human or animal subject with a condition caused or mediated by a virus of the Herpes family, which method comprises administering to said human or animal subject an effective amount of a compound of formula (I) or a physiologically acceptable salt or solvate thereof. The above compounds according to the invention and their pharmaceutically acceptable derivatives may be employed in combination with other therapeutic agents for the treatment of the above infections or conditions. Combination therapies according to the present invention comprise the administration of at least one compound of the formula (I) or a pharmaceutically acceptable derivative thereof and at least one other pharmaceutically active ingredient. The active ingredient(s) and pharmaceutically active agents may be administered simultaneously in either the same or different pharmaceutical formulations or sequentially in any order. The amounts of the active ingredient(s) and pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect. Preferably the combination therapy involves the administration of one compound according to the invention and one of the agents mentioned below.

Examples of such further therapeutic agents include agents that are effective for the treatment of viral infections or associated conditions such as (1 alpha, 2 beta, 3 alpha)-9-[2,3-bis(hydroxymethyl)cyclobutyl]guanine [(-)BHCG], oxetanocin-G(3,4-bis-(hydroxymethyl)-2-oxetanosyl]guanine), acyclic nucleosides (e.g. acyclovir, valaciclovir, famciclovir, ganciclovir, penciclovir), acyclic nucleoside phosphonates e.g. (S)-1-(3-hydroxy-2-phosphonyl- methoxypropyl)cytosine (HPMC), ribonucleotide reductase inhibitors such as 2- acetylpyridine 5-[(2-chloroaniiino)thiocarbonyl) thiocarbonohydra- zone, 3'-azido- 3'-deoxythymidine, other 2',3'-dideoxynucleosides such as 2^,,3'-dideoxycyt.dine, 2\3'-dideoxyadenosine and 2',3'-dideoxyinosine, 2',3'-didehydrothymidine, protease inhibitors such as N-tert-butyl-dehydro-2-[-2(R)-hydroxy-4-phenyl-3(S)- [[N-(2-quinolylcarbonyl)-L-asparginyl]butyl]-(4aS,8aS)-isoquinoline-3(S)- carboxamide, oxathiolane nucleoside analogues such as (-)-cis-1-(2- hydroxymethyl)-1 ,3-oxathiolan-5-yl)-cytosine (3TC) or cis-1-(2-(hydroxymethyl)- 1 ,3-oxathioian-5-yl)-5-fluoro-cytosine (FTC), 3'-deoxy-3'-fluorothymidine, 5- chloro^'.S'-dideoxy-S'-fluorouridine, (-)-cis-4-[2-amino-6-(cyclopropylamino)-9H- purin-9-yl]-2-cy-clopentene-1 -methanol, ribavirin, 9-[4-hydroxy-2-

(hydroxymethyl)but-1-yl]-guanine (H2G), tat inhibitors such as 7-chloro-5-(2- pyrryl)-3H-1 ,4-benzodiazepin-2(H)-one, or 7-chloro-1 ,3-dihydro-5-(1H-pyrrol-2- yl)-3H-1 ,4-benzodiazepin-2-amine, interferons such as -interferon, renal excretion inhibitors such as probenecid, nucleoside transport inhibitors such as dipyridamole; pentoxifylline, N-Acetylcysteine (NAC), Procysteine, -trichosanthin, phosphonoformic acid, as well as immunodulators such as interleukin II or thymosin, granulocyte macrophage colony stimulating factors, erythropoetin, soluble CD and genetically engineered derivatives, thereof, or non-nucleoside reverse transcriptase inhibitors such as nevirapine (BI-RG-587), loviride (-

APA) and delavuridine (BHAP), and phosphonoformic acid.

More preferably the combination therapy involves the administration of one of the above mentioned agents and a compound within one of the preferred or particularly preferred sub-groups within formula (I) as described above.

The present invention further includes the use of a compound according to the invention in the manufacture of a medicament for simultaneous or sequential administration with at least one other therapeutic agent, such as those defined hereinbefore.

The compounds according to the invention may be formulated for administration in any convenient way, and the invention therefore also includes within its scope pharmaceutical compositions for use in therapy, comprising a compound of formula (I) or a physiologically acceptable salt or solvate thereof in admixture with one or more physiologically acceptable diluents or carriers.

There is also provided according to the invention a process for preparation of such a pharmaceutical composition which comprises mixing the ingredients.

The compounds according to the invention may, for example, be formulated for oral, buccal, parenteral, topical or rectal administration.

Tablets and capsules for oral administration may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch or polyvinyl pyrrolidone; fillers, for example, lactose, microcrystalline cellulose, sugar, maize- starch, calcium phosphate or sorbitol; lubricants, for example, magnesium stearate, stearic acid, talc, polyethylene glycol or silica; disintegrants, for example, potato starch, croscarmellose sodium or sodium starch glycollate; or wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in the art. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example, sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxymethyl cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenated edible fats; emulsifying agents, for example, lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; or preservatives, for example, methyl or propyl p_- hydroxybenzoates or sorbic acid. The preparations may also contain buffer salts, flavouring, colouring and/or sweetening agents (e.g. mannitol) as appropriate.

For buccal administration the compositions may take the form of tablets or lozenges formulated in conventional manner.

The compound may also be formulated as suppositories, e.g. containing conventional suppository bases such as cocoa butter or other giycerides.

The compound according to the invention may also be formulated for parenteral administration by bolus injection or continuous infusion and may be presented in unit dose form, for instance as ampoules, vials, small volume infusions or pre- filled syringes, or in multi-dose containers with an added preservative. The compositions may take such forms as solutions, suspensions, or emulsions in aqueous or non-aqueous vehicles, and may contain formulatory agents such as antioxidants, buffers, antimicrobial agents and/or toxicity adjusting agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use. The dry solid presentation may be prepared by filling a sterile powder aseptically into individual sterile containers or by filling a sterile solution aseptically into each container and freeze-drying. By topical administration as used herein, we include administration by insufflation and inhalation. Examples of various types of preparation for topical administration include ointments, creams, lotions, powders, pessaries, sprays, aerosols, capsules or cartridges for use in an inhaler or insufflator or drops (e.g. eye or nose drops).

Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents and/or solvents. Such bases may thus, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as arachis oil or castor oil or a solvent such as a polyethylene glycol. Thickening agents which may be used include soft paraffin, aluminium stearate, cetostearyl alcohol, polyethylene glycols, microcrystalline wax and beeswax.

Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilising agents, dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of any suitable powder base, for example, talc, lactose or starch. Drops may be formulated with an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilising agents or suspending agents.

Spray compositions may be formulated, for example, as aqueous solutions or suspensions or as aerosols delivered from pressurised packs, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, 1 ,1 ,1 ,2,3,3,3-heptafluoropropane, 1 ,1 ,1 ,2- tetrafluorethane, carbon dioxide or other suitable gas.

Capsules and cartridges for use in an inhaler or insufflator, of for example gelatin, may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch.

Compounds of the invention may conveniently be administered in amounts of, for example, 0.01 to 50mg/kg body weight, suitably 0.05 to 25mg/kg body weight and preferably 1 to 25mg/kg body weight orally, one or more times a day. The precise dose will of course depend on the age and condition of the patient, the particular route of administration chosen, and the disease being treated.

The compounds of the formula (I) have useful duration of action.

The compounds of formula (I) and salts and solvates thereof may be prepared by the methodology described hereinafter, constituting a further aspect of this invention.

A process according to the invention for preparing a compound of formula (I) comprises:

(i)(a) condensation of a compound of formula (M) wherein R₁ is as defined above:

with a compound R₃S0₂Y where Y is a leaving group such as halogen e.g. chlorine and R₃ is as defined above or appropriately protected; or

(i)(b) condensation of a compound of formula (M') wherein R is as defined above:

with R₄R₅NH, wherein R₄, R₅ are as defined above; or

(ii) condensation of a compound of formula (N) wherein R₂ is as defined above:

with a compound R₉COOH, R₉COY, or R.Y' wherein Y is a leaving group such as halogen, e.g. chlorine, Y' is a halogen such as bromine or fluorine, R is optionally substituted heteroaryl with one to four heteroatoms and R₉ is as defined; or

(iii) converting one compound of the formula (I) into another compound of the formula (I); or

(iv) purifying one diastereomer of the compound of formula (I) from its racemic mixture by one or more of chromatography of a diastereomeric mixture, crystallisation or resolution using a chiral template;

and where desired or necessary converting a resultant free acid or base compound of formula I into a physiologically acceptable salt form or vice versa or converting one salt form into another physiologically acceptable salt form.

Process (i)(a)

The reaction with R₃S0₂Y is carried out in the presence of an organic base such as triethylamine in a suitable solvent such as dichloromethane at a temperature suitably between 0°C and ambient, or in the presence of an inorganic base such as sodium hydrogen carbonate in a vigorously-stirred two-phase system such as dichloromethane/water at a temperature suitably between 0°C and ambient.

Process (i)(b)

The reaction with R₄R₅NH with M' is carried out in the presence of an organic base such as DIPEA in a suitable solvent such as dichloromethane or DMF at a temperature suitably between 0°C and 60°C Process (ii)

Protection of the R₂ sulphur containing group may be required. Where R, is R₉CO, the R group may be incorporated by acylation carried out using the acyl chloride R₉COCI and lithium hexamethyldisilazide at a reduced temperature, suitably between 0°C and -78 °C in a solvent, suitably tetrahydrofuran. Alternatively, it may be carried out using the preformed mixed anhydride generated by treatment of the acid R_gCOOH with an appropriate acid chloride such as pivaloyl chloride in the presence of a base e.g. triethylamine or N,N- diisopropylethylamine in a solvent, suitably tetrahydrofuran, or using the preformed symmetrical anhydride. The reaction is carried out under similar conditions to those using the acid chloride.

The condensation reaction with R₉COOH is suitably carried out in the presence of a coupling agent such as 0-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) used with 1-hydroxybenzotriazole (HOBT) in the presence of an organic base such as N,N-diisopropylethylamine (DIPEA) and a solvent such as dichloromethane, tetrahydrofuran or dimethylformamide at a temperature of suitably between 0°C and ambient. It will be appreciated that as an alternative to using R₉COOH, acid derivatives such as the acid chloride, acid anhydride, or a mixed anhydride may be used. Reaction conditions will be modified accordingly, for instance where the reaction is with the acid activated as a mixed anhydride, with pivaloyl chloride and a base e.g. triethylamine or N,N-diisopropylethylamine in THF and where the acid chloride is used a suitable base is or lithium hexamethyldisilazide in THF.

Reaction with R₉Y' wherein Y' is bromine is the Goldberg variant of the Ullman reaction and is carried out in the presence of a copper catalyst such as copper(l)chloride in the presence of a base such as potassium carbonate in a high boiling inert solvent such as xylene at a suitably elevated temperature usually at reflux. The reaction is advantageously carried out in the presence of a solid-liquid phase transfer catalyst such as tris(3,6-dioxaheptyl)amine (TDA-1 ). Reaction with R₉Y' wherein Y' is fluorine is carried out in the presence of a base such as sodium hydride or lithium hexamethyldisilazide in a suitable solvent such as tetrahydrofuran at a temperature suitably between -78°C and ambient depending on the base used. The reaction with R₉COY and R₉S0₂Y is carried out in the presence of an organic base such as triethylamine in a suitable solvent such dichloromethane at a temperature suitably between 0°C and ambient.

Process (iii) Examples of typical interconversions include reducing a N0₂ group to NH₂, and alkenyl group to alkyl.

Process (iv)

Isolation of a single enantiomer may be achieved by conventional methods such as flash chromatography on silica gel; chiral chromatography (e.g. chiral HPLC) and crystallisation with a homochiral acid (e.g. tartaric acid) or base (e.g. norephedrine).

Physiologically acceptable base salts of the compounds of formula (I) may conveniently be prepared by treating a compound of formula (I) with a suitable base such as a bicarbonate, e.g. sodium bicarbonate, in the presence of a suitable solvent. Acid salts such as the hydrochloride, trifluoroacetate or tartrate may be prepared by treating a basic compound of formula (I) with the desired acid.

Preparation of intermediates

Certain intermediates useful in the preparation of compounds of formula (I) may be prepared using the following further processes:

(vi) Compounds of formula (M) above may be prepared by condensation of a compound of formula (P) wherein P, is a nitrogen protecting group,

with a compound R₉COOH, R₉COY, or R.Y' wherein Y is a leaving group such as halogen, e.g. chlorine, Y' is a halogen such as bromine or fluorine, R, is optionally substituted heteroaryl with one to four heteroatoms and Rg is as defined above, in a process analogous to (ii) above, followed by deprotection. (vi)(a) Compounds of formula (M¹) may be prepared by condensation of compounds of formula (M) above with a trialkylsilyl halide, e.g. trimethylsilyl chloride and a tertiary amine base, e.g. DIPEA, followed by treatment with sulphury! chloride.

(vii) N-protected compounds such as those of formula (XXIII) below may be prepared by condensation of a compound of formula (Q) wherein

P₂ and P₃ are different and represent nitrogen protecting groups o

sequentially with a base and then with a compound RY, or sequentially with a compound RY and then with a base, wherein Y is a leaving group such as those noted above and R represents C^ alkyl e.g. methyl. Further intermediates may then be formed by selective deprotection of P₂ and/or P₃ Similarly, where P₂ is a

nitrogen protecting group and R₁ is

, as in formula (R) below,

N-protected compounds of formula (M) above may be prepared by condensation of a compound of formula (R) sequentially with a base and then with a compound RY, or sequentially with a compound RY and then with a base, wherein Y is a leaving group such as those noted above and R represents C^ alkyl e.g. methyl.

These reactions are suitably carried out in the presence of a strong base, such as LiHMDS, in the presence of a solvent such as tetrahydrofuran, optionally containing a polar additive such as DMPU at a reduced temperature such as - 78° to 0°C.

(viii) Compounds of formula (N) above may be prepared by condensation of a compound of formula (S) wherein P is a nitrogen protecting group,

with a compound R₃S0₂Y where Y is a leaving group such as halogen e.g. chlorine and R₃ is as defined above, in a process analogous to (i) followed by deprotection.

Preparation of compounds of Formula (I)

In one example of this aspect of the invention, compounds of Formula (I) in which R₁ represents R₉CO as defined above may conveniently be prepared according to the methodology shown in the following general scheme 1 :

Scheme 1

(g)

NCOR_g

(XXVII) (h) (XXVI)

(XXVIa) Steps (a)

One possible route to the desired 2R.3S enantiomer of the intermediate (XX) is given below (steps p-u), wherein P, is a N-protecting group, preferably Boc (t- butyloxycarbonyl), P₂ is another N-protecting group, preferably Cbz (benzyloxycarbonyl) and R₁₂ is suitably a C^ straight or branched alkyl group e.g. ethyl or t-butyl.

(S) diaminobutyric acid

(S)

Step p The compounds of formula (XXXV)(S) are either known compounds or may be prepared in analogous manner to known compounds. The reaction is suitably carried out using PI FA (phenyl iodosylbis(trifluoroacetate) and a base such as pyridine in an aqueous solvent, such as aqueous THF, dioxan or acetonitrile. This is the method of Stansfield, C.F. Organic Preparations and Procedures Int., 1990, 22(5), 593-603.

Step q This protection reaction may be carried out in a conventional manner. For instance it is suitably carried out in a water miscible solvent such as THF, DMF or dioxan using N-(benzyloxycarbonyloxy)succinimide, benzyloxycarbonyl chloride, or any suitable source of the benzyloxycarbonyl group, with pH adjustment to alkaline with sodium carbonate.

As an alternative, step q¹, the compound of formula (XXXVII) can be prepared in conventional manner from (S) diaminobutyric acid.

Step r

This reaction is suitably carried out in two stages. Firstly, reacting at reduced temperature with N-methyimorpholine and then an alkyl chloroformate such as ethyl chloroformate, in an organic solvent such as DCM, dioxan or THF. Secondly, the intermediate product while in solution is reduced, suitably with sodium borohydride dissolved in a suitable solvent such as water, at reduced temperature, such as -20° to 10°C.

Step s

This oxidation reaction may be suitably carried out in any suitable manner, for instance using oxalyl chloride in DMSO and dichloromethane under nitrogen at reduced temperature, such as -30° to -70°C, followed by triethylamine. The intermediate (XXXIX) suitably is not isolated.

Step t This reaction is suitably carried out using a Wittig reagent such as a triphenylphosphorane R₁₂0₂CH=PPh₃, or may also be carried out using a phosphonate in a Wadsworth-Emmons reaction.

Step u This Michael addition reaction is suitably carried out using lithium bis(trimethylsilylamide) or other suitable strong base in a suitable organic solvent such as THF, ether or toluene, and preferably a complexing agent such as tetramethylethylenediamine is also present. Alternatively, for step (a), an achiral preparation may be employed and the mixture of enantiomers (XX) may be resolved so that the required 2R.3S- enantiomer is brought through step (b) and the following steps chirally. Any suitable resolving agent, preferably (+) di-p-toluoyl-tartaric acid ((+)-DPTTA) followed by typically two recrystallisations suitably from ethanol, is used to give the 2R,3S-enantiomer as the tartrate, (XX)2R,3S.

(XX)2R,3S

As a further alternative, the racemic mixture (XX) may be processed through steps (b) to (h) and the enantiomeric separation carried out at a later stage. Another alternative preparation of intermediate (XX) is described with reference to scheme 3 below.

Step (b)

Deprotection carried out with addition of acid such as trifluoroacetic acid followed by cyclocondensation with an alkyl Grignard reagent, suitably 'ButylMgCI in a solvent, suitably tetrahydrofuran, at a temperature between -20°C and ambient.

Step (c)

Reprotection is carried out in a conventional manner. When P₃ is Boc, this is suitably achieved with Boc₂0 in a solvent suitably tetrahydrofuran at a suitable temperature such as -78°C in the presence of a base such as lithium hexamethyldisilazide or sodium hydroxide, or with Boc₂0 in a solvent suitably acetonitrile at a suitable temperature such as ambient in the presence of an acylation catalyst such as dimethylaminopyridine (DMAP).

Step (d) Alkylation reaction, where R is other than hydrogen, which may be carried out by treating sequentially with a base and then with a compound RY, or sequentially with a compound RY and then with a base, wherein Y is a leaving group such as a halogen and R is C .₃ alkyl. This reaction may suitably be carried out using methyl iodide when it is desired to introduce a methyl group as the R substituent in Formula (I), as is depicted in scheme 1. Where R represents hydrogen, this step is omitted. The following steps (e) to (h) are then carried out as described.

It has been found that where the alkylation reaction is carried out with Mel, in tetrahydrofuran as solvent at -78°C, a predominance of the desired α-methyl stereoisomer over the β-methyl results, in a ratio of at least 50:1.

Step (e) The deprotection is carried out by conventional means, e.g. by addition of acid such as trifluoroacetic acid.

Step (f)

Where R is R₉CO, the R₁ group may be incorporated by acylation carried out using the acyl chloride R₉COCI and lithium hexamethyldisilazide at a reduced temperature, suitably -78 °C in a solvent, suitably tetrahydrofuran. Alternatively, it may be carried out using the preformed mixed anhydride generated by treatment of the acid R₉COOH with an appropriate acid chloride such as pivaloyl chloride in a solvent, suitably tetrahydrofuran. The reaction is carried out under similar conditions to those using the acid chloride. Where R is R₉NHCO, the reaction to incorporate R may be carried out with the appropriate isocyanate, RgNCO, in the presence of less than 1 mol equivalent of a suitable base such as sodium hydride, preferably 0.3mol equivalents, in a solvent suitably tetrahydrofuran at ambient temperature. In instances where a highly reactive isocyanate such as chlorosulphonyl isocyanate or chloroacetyl isocyanate is used, the base may be omitted.

Step (g)

Deprotection, which can be carried out by conventional means, e.g. where P₂ is Cbz by hydrogenation in the presence of a palladium catalyst in a suitable solvent such as propan-2-ol. The product is preferably isolated as a salt, such as the hydrochloride salt.

Step (h) Where R₂ is -S0₂R₃, as set out in formula (I), the group -S0₂R₃ may be introduced in one step, by use of the procedure described for process (i) above.

Step (j)

Conversion of (XXVI) to the chlorosulphonyl analogue (XXVIa) may be carried out by treatment of (XXVI) with a suitable silylating agent such as trimethylsilyl chloride and a suitable tertiary amine base such as DIPEA or triethylamine followed by treatment with sulphuryl chloride in a suitable inert solvent such as dichloromethane at a temperature between 0°C and ambient.

Step (k)

Condensation of (XXVIa) with R₄R₅NH may be carried out in the presence of a suitable base such as DIPEA or triethylamine in a suitable inert solvent such as dichloromethane at a temperature between 0°C and 60°C.

In a second embodiment of this aspect of the invention, compounds of formula (I) wherein R₁ represents optionally substituted heteroaryl may conveniently be prepared according to the methodology shown in the following general scheme 2, in which steps corresponding to those discussed above in relation to scheme 1 are correspondingly labelled: Scheme 2

(XXXII) Step (I)

The condensation may be carried out according to the procedure described for process (i) above using R₃S0₂Y.

Step (m)

This coupling reaction may be carried out according to the procedure described for process (ii) starting with the appropriate haloheteroaromatic compound, R^' and using for example the Goldberg variant of the Ullman Reaction. It will be appreciated that the compounds of formula (XXXV) below may also be used for the synthesis of compounds of formula I, specifically compounds of formula (XXXII) above, by protection of the pyrrolidine nitrogen with an acid labile protecting group, such as 4-methoxybenzyloxycarbonyl (Moz), introduction of the group R^ deprotection by conventional means and introduction of the group -S0₂R₃ as described above.

In cases where R₃ is R₄R₅N, compounds of formula (XXXII) may also be prepared from another compound of formula (XXXII) wherein R₃ is R₄NH by alkylation using a suitable alkylating agent, such as t-butyl bromoacetate or allyl bromide, in the presence of a suitable base, such as caesium carbonate, in a suitable solvent, such as acetonitrile and/or dichloromethane.

H.

The compound of formula (XXXV) may be obtained, for example, by deprotection of the compound of formula (XXVIII) using the procedure of Step (e) described above.

In a third example of this aspect of the invention, compounds of formula (I) wherein R represents methyl may conveniently be prepared according to the methodology shown in the following general scheme 3. In addition, Scheme 3 shows a novel alternative synthesis of intermediate (XX). Scheme 3

L-methionine

(o)|

C

(w)|

Step (n)

This is a conventional protection reaction which, in the case when P₂ represents BOC, may be performed by reacting with (BOC)₂0 in the presence of base (e.g. NaOH) in a polar solvent system such as dioxan/water.

Step (o)

This conversion may be performed on treatment with ammonium bicarbonate in the presence of a suitable solvent such as pyridine/DMF and in the presence of (BOC)₂0 or suitable equivalent.

Step (v)

This is a conventional protection reaction which, in the case when P, represents CBZ, may be performed by reaction with nBuLi followed by CBZ-CI in the presence of an inert solvent such as THF below -50 °C.

Step (w)

This reaction may be performed in two stages, firstly by treatment with RX where RX is a compound (e.g. Mel, benzyl iodide or Me₂S0₄) capable of converting sulphur in the SMe moiety to sulphonium in a suitable solvent, e.g. propanone or acetonitrile. Generally R will represent alkyl or aralkyl and X will represent halide, especially iodide, or sulphate. Protection of the amide is convenient, although not essential, for this reaction. Secondly, the ring closure reaction may be performed by treatment with Dowex 2 x 8 400 mesh OH^" resin in a suitable solvent, e.g. MeCN. Alternatively, the ring closure may be performed by treatment with potassium carbonate in a suitable solvent, e.g. MeCN.

Step (x)

Following deprotection, which may be performed in a conventional manner, for example, a BOC protecting group may be removed by treatment with HCI, e.g. in dioxan, the amine may be treated with a trifluoroacetic acid alkyl ester (e.g. the methyl ester) or trifluoroacetic anhydride in the presence of a suitable base e.g. N-methylmorpholine, then addition of a reducing agent e.g. lithium borohydride, followed by treatment with concentrated sulphuric acid in the presence of an alkyl alcohol e.g. ethanol solvent, gives the ether (XLV).

Step (y)

The reaction of compounds of formula (XLV) and the silyl ketene acetal derived from methyl propionate takes place in the presence of a Lewis acid e.g. BF₃.OEt₂ and an inert solvent e.g. dichloromethane or MeCN. The group "alkyl" in OSi(alkyl)₃ generally represents C^alkyl, suitably methyl or isopropyl. Preferred OSi(alkyl)₃ is OSi(i-Pr)₃. The use of variants in which OMe is replaced by OSi(alkyl)₃ is also envisaged.

Step (yy)

The compound of formula (XLV) is reacted with diethyl malonate in the presence of a Lewis acid e.g. tin (IV) chloride and an inert solvent e.g. dichloromethane or acetonitrile.

Step (yyy) Hydrolysis of compounds of formula (XLVIa) is carried out by treatment with a suitable base e.g. sodium hydroxide in a suitable solvent e.g. aqueous ethanol at a temperature between 0°C and ambient. Subsequent decarboxylation and esterification may be carried out in a suitable solvent such as ethanol at reflux in the presence of an acid such as sulphuric acid.

Step (z)

This deprotection reaction will take place on treatment with base, such as potassium carbonate.

Step (zz)

This ring closure reaction may be performed on treatment with an alkyl Grignard reagent (e.g. t-butylmagnesium choride) in an inert solvent such as THF in the presence of tetramethylethylenediamine at a temperature of -20°C to 25°C. Compounds of formula (XLVIII) are a subset of compounds of formula (XXIV) in scheme 1 and can be carried forward through steps (f), (g) and (h) of that scheme to give compounds of formula (I).

As an alternative to the synthesis shown in Scheme 3, the starting material may be D, L-methionine and the racemic R,S compound of formula (XLIV) thus produced may be separated by dynamic resolution with a homochiral preparation of an appropriate chiral acid, e.g. (-) di-p-toluoyl-tartaric acid, to give the desired S isomer (90% yield). ) DPTTA

(R

reprotect

(S) XLIV

In a further aspect, the present invention provides the use of a compound of formula (XXII) in the synthesis of a compound of formula (I). Further, the present invention provides a method of making a compound of formula (I) comprising alkylating a compound of Formula (XXII) at the carbon atom adjacent to the lactam carbonyl group.

(XXII)

In a further aspect, the present invention provides the use of a compound of formula (L)

wherein R is C,.₃alkyl, P₃ is a protecting group such as CBZ and R₁₈ is C^ alkyl, and its use in the preparation of compounds of formula (I) in which R is C^ alkyl. An example of such a compound is the compound of formula (XLVII) and an example of its use in the preparation of compounds of formula (I) can be seen in Scheme 3 above.

In a further aspect, the present invention provides a compound of formula (XXIII)

in the synthesis of a compound of formula (I). Further, the present invention provides a method of making a compound of formula (I) comprising removal of the protecting group P₂ in the compound of formula (XXIII) followed by condensation with R₃S0₂Y at nitrogen of the pyrrolidine ring to introduce the group R₂. Further, the present invention provides a method of making a compound of formula (I) comprising removal of the protecting group, P₃ in the compound of formula (XXIII) followed by reaction at the nitrogen of the lactam ring to introduce the group R

ABBREVIATIONS

Boc t-butyloxycarbonyl

CBZ Benzyloxycarbonyl

DCM Dichloromethane

(Boc)₂0 Di-tert-butyldicarbonate

Et₃N Triethylamine

THF Tetrahydrofuran

TFA Trifluoroacetic Acid

LiHMDS Lithium bis (trimethylsilyl)amide

DMF Dimethylformamide

TBTU 0-(1 H-Benzotriazol-1 -yl)-N,N,N\N'- tetramethyluronium tetrafluoroborate

HOBT 1 -Hydroxybenzotriazole

DIPEA N,N-Diisopropylethyiamine

Moz 4-Methoxybenzyloxycarbonyl TDA-1 Tris[2-(2-methoxyethoxy)ethyl]amine

IMS Industrial methylated spirit

Dansyl 5-dimethylamino-naphthalene-1-sulfonyl

The following non-limiting examples illustrate the present invention.

Synthetic examples:

Preparative hplc was carried out at room temperature on a Gilson X233

Autoprep system with a Supelcosil SP0580 LC-ABZ+PLUS column (10 cm x

21.2 mm ID, flow rate 4 mL min^"1) using the following protocols, wherein A denotes acetonitrile containing 0.05% of 98% formic acid and B denotes water containing 0.1% of 98% formic acid: fcaH : 20%A:80%B to 40%A:60%B linearly over 20 mins, then 40%A:60%B for

10 mins then 100%A for 10 mins ( flow rate 4 mL min ¹) fcal2: 30%A:70%B to 60%A:40%B linearly over 20 mins, then 60%A:40%B for 5 mins then 100%A for 20 mins fcal3: 60%A:40%B to 100%A linearly over 20 mins, then 100%A for 15 mins

LC-MS was carried out at room temperature using a Micromass series II mass spectrometer with a Hewlett-Packard HP1100 Autoinjector and a 3.3 cm x 4.6 mm ID 3 μM ABZ+PLUS column using the following protocols, wherein C denotes 10mM ammonium acetate in water containing 0.1% of 98% formic acid and D denotes acetonitrile containing 0.05% of 98% formic acid: Method A: 0.00-0.70 mins (100%C), 0.70-4.20 mins 100%C to 100%D linearly, 4.20-5.30 mins (100%D), 5.30-5.50 mins 100%D to 100%C linearly (flow rate 3 mL min^"1 )

Method B: 0.00-0.70 mins (100%C), 0.70-4.20 mins 100%C to 100%D linearly, 4.20-7.70 mins 100%D, 7.70-8.00 mins 100%D to 100%C linearly (flow rate 1 mL min^"1 )

Synthesis of intermediates: Intermediate 1

1 -{3-f(Benzyloxy-carbonyl)-amino]-1 -hydroxymethyl-propyl}-carbamic acid, tert- butyl ester

A solution of compound Nα-BOC,Nγ-CBZ-2,4-Diaminobutyric acid (3.198g) in tetrahydrofuran (44ml, dry) was cooled to -10°C under nitrogen, 4- methylmorpholine (1.0ml) was added followed by ethylchloroformate (0.868ml). After stirring for 8 mins sodium borohydride (1.03g) was added in one portion followed by methanol (88ml) over a period of 11 mins at 0°C. The mixture was stirred at ca 0°C for an additional 11 mins before 1 M hydrochloric acid (18ml) was added. The mixture was evaporated under reduced pressure and the aqueous residue was extracted with ethyl acetate. The organic layer was separated and washed with 1 M hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and water, then dried (magnesium sulphate), evaporated under reduced pressure and some of the residue (1.8g from 2.87g) was purified by chromatography (Merck 7734) using cyclohexane:ethylacetate (3:2) as eluent to give the title compound (1.6g) : t.l.c. (1 :1 cyclohexane : ethyl acetate) Rf 0.23 ir (CHBr₃) 3432, 1704cm-¹.

Intermediate 2

6-Benzyloxycarbonylamino-4-fetf-butoxycarbonylamino-hex-2E-enoic acid ethyl ester

A solution of dimethyl sulphoxide (6.82ml) in dry dichloromethane (135ml) was stirred under N2 and cooled (dry ice/acetone) to - 72°C. Oxalyl chloride (7.4ml) was added dropwise over 10 minutes (temp kept in the range - 60 → 65°C) and the reaction was stirred for 15 minutes. A solution of the alcohol, Intermediate 1 , (12.6g) in dry dichloromethane (135ml) was added over 20 minutes (temp kept in the range -60°-63°C) and the reaction mixture then stirred for 20 minutes by which time the temperature had risen to -52°C. Triethylamine (53.7ml) was added dropwise over 10 minutes followed by the immediate addition of the Wittig reagent (19.3g). The cooling bath was removed and the internal temperature allowed to rise to 17°C. The reaction mixture was poured into ether (400ml) and brine (400ml). The organic phase was separated and the aqueous phase extracted with ether (2x100ml). The combined organic phases were dried (MgS04) and evaporated under reduced pressure to give a tan oil (36.22g). This was purified by flash column chromatography (Merck 9385 silica eluting with 40% ethyl acetate in cyclohexane) to give the title compound (15.71g) as an oil: Η nmr (CDCI₃): δ 7.40-7.30 (5H, m), 6.86 (1 H, dd), 5.93 (1 H, dd), 5.42-5.28 (1 H, br), 5.12 (2H, ABq), 4.72-4.60 (1 H, m), 4.50-4.32 (1 H, m), 4.19 (2H, q), 3.60- 3.30 (1 H, m), 3.15-2.98 (1 H, m), 2.00-1.80 (1 H, m), 1.65-1.50 (1 H, m), 1.45 (9H, s) and 1.28 (3H, t), Rf 0.45 (2:3 ethyl acetate /cyclohexane)

Intermediate 3 rel-(2R,3S)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1- carboxylic acid benzyl ester

Intermediate 2 (12.2g) was suspended in dry toluene (175ml) with stirring under N2- Tetramethylethylenediamine (1.1ml) was added followed by lithium bis- (trimethylsilyl)amide (1.0M in hexanes, 7.6ml). On completion of the addition a solution had formed. The reaction mixture was stirred for 15 minutes and then poured into ethyl acetate (300ml) and saturated aqueous ammonium chloride (300ml). The organic phase was separated and the aqueous phase extracted with ethyl acetate (2x50ml). The combined organic extracts were washed with brine (150ml) and the aqueous phase extracted with ethyl acetate (2x25ml). The combined organic extracts were dried (MgS04) and evaporated under reduced pressure to give a tan oil (12.86g) which was filtered through a plug of silica gel using ethyl acetate/cyclohexane (2/3) as eluant to give a crude mixture including title compound (10.74g) as an oil. This oil was purified further by flash column chromatography on silica gel. Elution with ethyl acetate/cyclohexane (2/3) gave the title compound, as a solid (8.49g, 69.7%). A small sample of the title compound was crystallised from ether to give a white solid: ¹H nmr (CDCI₃): δ 7.40-7.30 (5H, m), 5.12 (2H, s), 4.72-4.53 (1H, m), 4.20-3.95 (4H, m), 3.65- 3.40 (2H, m), 2.95-2.65 (1 H, m), 2.60-2.40 (1 H, m), 2.25-2.10 (1 H, m), 1.92-1.75 (1 H, m), 1.40 (9H, s) and 1.30-1.15 (3H, m). Rf 0.8 (1 :1 , ethyl acetate/cyclohexane)

Intermediate 4 trans-3-Amino-2-ethoxycarbonylmethyl-pyrrolidine- -carboxylic acid benzyl ester To the Boc-protected Intermediate 3 (246.6gm, 1eq, 0.607mol) was added trifluoroacetic acid (25eq, 15.18mol, 1731gm, 1169ml) at room temperature. After stirring for one hour the solution was evaporated and the residue azeotroped twice with toluene (300ml). The resulting oil was dissolved in ethyl acetate (2500ml) and washed with 2M sodium hydroxide (1x800ml + 3x300ml), water and brine, dried (MgS0₄) and evaporated to give the title compound as a golden oil, 168.9gm, after high vacuum. Mass spec 613 [2M+H]⁺, 307 [MH]⁺.

Intermediate 5

(2R,3S)-3-amino-2-ethoxycarbonylmethyl-pyrrolidine-1 -carboxylic acid benzyl ester (2S,3S)-bis-(4-methyl-Q-benzyloxy)-succinate salt.

A solution of Intermediate 4 (168.9gm, 0.55mol) was dissolved in ethanol (2500ml) and was added to a solution of (+) di-0-para-toluyl-D-tartaric acid (ex Fluka) (213gm, 0.55mol) in ethanol (2500ml) and the solution allowed to stand overnight. The resulting solid was collected by filtration and washed with ethanol and then recrystallised from boiling ethanol (~3500ml) to give the title compound as a white solid, 93.7gm. A further recrystallisation gave a white solid, mp 184-185°C. Chiral HPLC (Chiracel OJ column, eluent system ethano heptane; 3:7; flow rate = 1ml/min, , λ = 215nm). Retention time = 7.7 min, 97.5 %ee.

Intermediate 6

(2R,3S)-3-amino-2-ethoxycarbonylmethyl-pyrrolidine-1 -carboxylic acid benzyl ester.

The Intermediate 5 (131.8 gm, 190mmol) was suspended in an 1 :1 mixture of water: ethyl acetate (1500ml), and solid potassium carbonate added (63gm, 457mmol). After fifteen minutes the phases were separated and the aqueous phase extracted with ethyl acetate (3x200ml). The organic portions were combined and washed with water and brine, dried (MgS0₄) and evaporated to give the title compound as a colourless oil, 57.8gm.

Mass spec 613 (100%) [2M+H]⁺, 307(83%) [MH]⁺. [α]_D = -11.3° (c=1.33, MeOH)

Alternative synthesis of intermediate 6 (via Intermediates 61 and 62): Intermediate 6

(2R,3S)-3-amino-2-ethoxycarbonylmethyl-pyrrolidine-1 -carboxylic acid benzyl ester. Intermediate 62 (11.3g, 23.84mmol) was dissolved in ethanol (40ml) and treated with sodium hydroxide solution (10N, 7.87ml, 78.7mmol). After standing at room temperature for 20h the solvent was removed in vacuo and the residue azeotroped with cyclohexane (2x100ml). The resulting white solid was dried in vacuo for 24h and then suspended in ethanol (130ml) and treated with concentrated sulphuric acid (3.17ml). The mixture was heated at reflux for 17h. More concentrated sulphuric acid (0.64ml) was added and the mixture refluxed for a further 7h. After cooling to room temperature the ethanol was removed in vacuo and the residue partitioned between ethyl acetate (200ml) and saturated sodium bicarbonate solution (200ml). The organic phase was separated and the aqueous extracted with ethyl acetate (200ml). The combined organic extracts were washed with brine (100ml), dried (MgS04) and concentrated to give the title compound as a purple oil (6.4g)

Identical to intermediate 7 in PG3207 LC-MS (method A) Peak at 2.22 mins gave m/e 307 (MH)⁺

Intermediate 7

(3aS,6aR)-5-Oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1 -carboxylic acid benzyl ester

To the Intermediate 6 (52.9gm, 173mmol) in tetrahydrofuran (550ml) in an ice- salt bath was added a solution of tert butyl magnesium chloride (554ml of a 1M solution in tetrahydrofuran, 554mmol), keeping the temperature <1°C. The mixture was warmed to room temperature over 1 hr 15min, then quenched with saturated ammonium chloride whilst cooling in an ice bath. The phases were separated and the aqueous phase extracted with ethyl acetate. The combined organics were washed with water and brine, dried (MgS0₄) and evaporated and to give the title compound as a cream solid, 43.5gm. A portion of the solid was purified by flash column chromatography over silica gel ( Merck 9385) using ethyl acetate as the eluting solvent, to give the title compound as a white solid, mp 157-159°C.

[α]_D = -68.4° (c=1.28, MeOH) Intermediate 8

(3aR,6aS)-2-Oxo-hexahydro-pyrrolo[3,2-blpyrrole-1 ,4-dicarboxylic acid 4-benzyl ester 1 -tert-butyl ester

To the Intermediate 7 (43.2gm, 166mmol) in tetrahydrofuran (1200ml) at -72°C under a nitrogen blanket was added lithium bis (trimethylsilyl)amide (216ml of a 11vl solution in tetrahydrofuran, 216mmol) dropwise, keeping the temperature <- 71 °C. After ten minutes a solution of di-tert-butyldicarbonate (54.3 gm, 249mmol) in tetrahydrofuran (350ml) was added, keeping the temperature <- 71 °C. The reaction was stirred at -73°C for two and a half hours and then quenched with saturated ammonium chloride. Then the mixture was allowed to warm to room temperature, water was added and the phases separated. The aqueous phase was extracted with ethyl acetate and the combined organics were washed with water and brine, dried (MgS0₄) and evaporated to give the title compound as an orange-red semi solid. Pure material could be obtained as a pale cream solid by trituration under diethyl ether. Yield 42.5gm. A portion of the solid was recrystallised from boiling diethyl ether to give the title compound as a white solid, mp 101-103°C. [α]_D = -45.6° (c=1.13, MeOH).

Intermediate 9

(3S,3aR,6aS)-3-Methyl-2-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1 ,4-dicarboxylic acid 4-benzyl ester 1 -tert-butyl ester

The Intermediate 8 (606mg, 1eq, 1.68mmol) was dissolved in tetrahydrofuran (6ml) and cooled, under nitrogen, to -75°C. Lithium hexamethyldisilazide (1.3 eq, 2.2ml of a 1 M solution in tetrahydrofuran) was added, keeping the temperature below -70°C. After 10 minutes methyl iodide was added (17eq, 28.9mmol, 1.8ml). After stirring for a further 45 minutes the reaction was quenched with saturated aqueous ammonium chloride and then allowed to warm to room temperature. Water was added and then the aqueous phase was extracted with ethyl acetate, and the combined organic phase was washed with water and brine, dried (MgS0₄) and evaporated to give a golden oil. Purification by flash column chromatography over silica gel ( Merck 9385) using cyclohexane:ethyl acetate (3:1 )as the eluting solvent system, afforded the title compound as a white foam, 526mg. Mass spec 275 [M-100+H]⁺ [α]_D = -88.6° (c=1.1 , MeOH).

Intermediate 10

(3aS,6S,6aR)-6-Methyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylic acid benzyl ester

To the Intermediate 9, (486mg, 1eq, 1.3mmol) was added trifluoroacetic acid (60eq, 6ml) and the mixture stirred at room temperature for 40 minutes, then evaporated to give a brown oil. This was dissolved in ethyl acetate (6ml) and washed with saturated sodium bicarbonate solution (2x3ml), water (3ml) and brine (3ml), dried (MgS0₄) and evaporated to give the title compound as a pale beige solid 340mg. The solid was recrystallised from boiling diethyl ether to give a white solid (202mg), mp 112-113°C.

Chiral HPLC (Chiral Pak464 column, eluent system propan-2-ol:heptane; 2:25; flow rate = 1 ml/min). Retention time = 20.91 min, 96.00 %ee.

Intermediate 11

(3aS,6S,6aR)-4-Cyclopropanecarbonyl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- frlpyrrole-1 -carboxylic acid benzyl ester

To a solution of the Intermediate 10, (992mg, 1eq, 2.75mmol) in dry THF (7ml) at -78°C under nitrogen was added lithium hexamethyldisilazide (3.3ml of a 1M solution in tetrahydrofuran, 1.2eq, 3.3mmol), keeping the temperature below -

70°C. The solution was kept at -78°C. for 10 mins, then at 0°C for 10 mins, recooled to -78°C and cyclopropanecarbonyl chloride (0.75ml, 3eq, 8.25mmol) was added and the reaction mixture was the stirred at -78°C. for 55 minutes. The reaction mixture was quenched with saturated aqueous ammonium chloride

(25ml) and then allowed to warm to room temperature. Water was added (20ml) and then the aqueous phase was extracted with ethyl acetate (100ml), and the combined organic phase was washed with water (30ml) and brine(30ml), dried

(MgS0₄) and evaporated to give a yellow oil. Purification by flash column chromatography over silica gel ( Merck 9385) using cyclohexane:ethyl acetate

(4:1 )as the eluting solvent system afforded the title compound as a pale yellow gum, 334mg. Mass Spec: 343 [MH]⁺

Intermediate 12 (3aS,6S,6aR)-1-Cyclopropanecarbonyl-3-methyl-hexahydro-pyrrolo[3,2-b1pyrrol- 2-one hydrochloride

A solution of Intermediate 11 , (330mg, 0.96mmol) in isopropanol (30ml) was added to the catalyst (119mg, 10% palladium on activated carbon with 50% water, Degussa type E101 NE/W) under nitrogen and the resulting mixture stirred vigorously under an atmosphere of hydrogen for 2.75hours. The catalyst was filtered off under an atmosphere of nitrogen and a 1 M solution of hydrogen chloride in diethyl ether (1 ml, 1eq, 1 mmol) was added to the filtrate. Evaporation of the solvent gave the title compound as a colourless gum, 175mg. Mass Spec: 209 [MH]⁺

Intermediate 13 rel-(3aS,6S,6aR)-1-Cyclopropanecarbonyl-3-methyl-hexahydro-pyrrolo[3,2- b]pyrrol-2-one hydrochloride Prepared analogously to intermediate 12, omitting the resolution step with (+) di- 0-para-toluyl-D-tartaric acid. Mass Spec: 209 [MH]⁺

Intermediate 15

(3aS,6S,6aR)-4-(cis-2,3-Dimethyl-cyclopropanecarbonyl)-6-methyl-5-oxo- hexahydro-pyrrolof3,2-b]pyrrole-1 -carboxylic acid benzyl ester

To a solution of c.s,c/^'s-bis(2,3-dimethylcyclopropanecarboxylic) acid ( 678mg, 5.95 mmol ) in anhydrous tetrahydrofuran ( 20 mL ) stirred under nitrogen at -13° was added methanesulphonyl chloride ( 250μL, 3.23 mmol ) followed by a solution of triethylamine ( 1.5mL, 10.8mmol ) in anhydrous tetrahydrofuran ( 10mL ) dropwise over 15 min at -12 to -14°. The resulting suspension was stirred below -11° for 1 h and then allowed to warm up to 20° over 2.5h. Solid was filtered off and bed-washed with ether and the combined filtrates were evaporated. The residue was partitioned between ether ( 2x 50mL ) and ice-cold saturated aqueous sodium bicarbonate solution ( 25mL ) . The combined organic phases were washed with water ( 20mL ) and saturated brine ( 20mL ), dried over magnesium sulphate and evaporated to give c/s,c/s-bis(2,3-dimethyl- cyclopropanecarboxylic) anhydride ( 543mg ) as an oil which crystallised. [MNH₄] ⁺ 228.

To a solution of the Intermediate 10 ( 600mg, 2.19 mmol ) in anhydrous tetrahydrofuran ( 8 mL ) stirred under nitrogen in an ice and IMS bath was added 1M lithium hexamethyldisilazide ( 2.4mL; 2.4 mmol ) slowly. The yellow solution was stirred for a further 20 min and then a solution of c.s,c/s-bis(2,3-dimethyl- cyclopropanecarboxylic) anhydride ( 510mg, 2.42mmol ) in anhydrous tetrahydrofuran ( 7 mL ) was added slowly. The resulting solution was stirred for 1h under nitrogen in an ice and IMS bath and then poured into saturated aqueous ammonium chloride solution ( 50 mL ). The solution was extracted with ethyl acetate ( 2x 50mL ) and the combined organic phases were washed sequentially with saturated aqueous sodium bicarbonate solution ( 20mL ), water ( 20mL ) and saturated brine ( 20mL ), dried over magnesium sulphate and evaporated to give an oil which crystallised ( 1.09g ) . The crude product was purified on a 10g Bond Elute^® Silica cartridge eluting with cyclohexane / dichloromethane to give the the title compound, ( 785mg). LCMS: 371 [MH]⁺, Retention time = 3.45 min, using Gilson Supelcosil LC ABZplus column, eluent system A (H₂0, 0.1% Formic acid, 10mmol Ammonium Acetate) B (MeCN, 0.05% Formic acid):Gradient 100%A 0.7mins, 100%A-100% B 3.5mins,100% B 3.5mins, 100%-0%B 0.3mins; flow rate = 1 ml/min.

Intermediate 16

(3S,3aR,6aS)-1-(cis-2,3-Dimethyl-cyclopropanecarbonyl)-3-methyl-hexahydro- pyrrolof3,2-b1pyrrol-2-one hydrochloride

A solution of the Intermediate 15 ( 890mg, 2.4 mmol ) in isopropanol ( 70mL ) containing 1M ethereal HCI ( 2.5mL , 2.5 mmol ) was hydrogenated in the presence of the catalyst (0.2g, 10% palladium on activated carbon with 50% water, Degussa type E101 NE/W) for 1.5h. The catalyst was filtered off , 1M ethereal H CI ( 0.2mL; 0.2mmol) was added and the solvent was evaporated off. Toluene ( 30mL ) was added and evaporated, and the resulting solid was triturated under ether ( 40mL ) . Ether ( 50mL ) was added and evaporated off to give the title compound (670mg) as a white solid.

LCMS: 237 [MH]⁺, Retention time =2.03 min, using the same conditions as in Intermediate 15.

Intermediate 17

(3^"S,3aR,6aS)-3-Methyl-2-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1 -carboxylic acid tert-butyl ester hydrochloride

Prepared in a similar manner to Intermediate 14 from Intermediate 9. The title compound , was obtained as a white solid in 93% yield and was used in the next reaction without further purification. Mass Spec. : 141 [M-Boc]⁺, 241 [MH]⁺, 481 [2MH]⁺

Intermediate 18 (3S,3aR,6aS)-3-Methyl-hexahydro-pyrrolo[3,2-blpyrrol-2-one hydrochloride

To the Intermediate 17, (8.61g, 31.1 mmol) in dry dichloromethane (20mL) was added trifluoroacetic acid (20mL, 260mmol, 8 eq.). The solution was stirred at room temperature for 2 hours. The solvents were then evaporated and the resulting brown gum was azeotroped with toluene (4X 50 mL). The title compound , was obtained in quantitative yield as a brown gum which was used in the next reaction without further purification. Mass Spec: 141 [MH]⁺ parent amine.

¹H nmr (d⁶-DMSO) : δ 9.18-8.80 (2H,brd.), 7.30-7.08 (1 H,m), 3.92-3.20 (4H,m), 2.60 (1 H,m), 2.26-2.10 (1 H,m), 1.75 (1 H,m), 1.09 (3H,d) ppm.

Intermediate 19

(3aS,6S,6aR)-6-Methyl-5-oxo-hexahydro-pyrrolθι^r3,2-b]pyrrole-1-carboxylic acid 4-methoxy-benzyl ester

Intermediate 18 (12.01g, 31.1 mmol of trans-lactam + 57 mmol TFA) and triethylamine (11.2 mL, 80.43 mmol, 2.6 eq. w.r.t. trans-lactam) were dissolved in water (40mL). A solution of 2-(4-methoxybenzyloxycarbonyioxyimino)-2- phenylacetonitrile (MOZ-ON) (7.45g,31 mmol, 0.8 eq.) in 1,4-dioxane (60mL) was added to the stirred mixture. After 18 hours it was diluted with water (140mL) and extracted with ethyl acetate (2 X 100mL). The aqueous phase was acidified to pH2 with solid citric acid and then saturated with solid sodium chloride. It was then further extracted with ethyl acetate (3 X 60mL). The combined organic extracts were washed with water (50mL) and sat. brine (50mL), dried over MgS0₄ and the solvent was evaporated to give a brown oil. Trituration under ether caused precipitation of a white solid which was collected by filtration washed with ether and dried in vacuo at room temperature to give the title compound (5.37g).

LCMS: 305.2 [MH]⁺, Retention time = 3.76 min, using the same conditions as in Intermediate 15.

Intermediate 20 (3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -carboxylic acid 4-methoxy-benzyl ester

To the Intermediate 19, (1.04g.3.42mmol,1eq.) was added 2- bromobenzothiazole¹ (1.17g,5.47mmol,1.6eq.), potassium carbonate (0.768g,5.56 mmol, 1.6 eq.), copper (I) chloride (0.345g,3.49mmol,1eq.), TDA-1 (0.330mL,1.03 mmol,0.3eq.) and xylene (55mL). The reaction mixture was refluxed under a Dean-Stark head for 7.5 hours. After leaving to cool, the brown solids were filtered off and washed with ethyl acetate (30mL). The combined filtrate and washings were washed with water (100mL) and brine (100mL) and dried over MgS0₄. The solvent was evaporated to give a brown gum (1.99g). The title compound, was purified by flash column chromatography over silica gel (Merck 9385) using cyclohexane:ether and was obtained as a white solid (0.95g). Mass Spec. : 438 [MH]⁺ Ref 1. M.P. Doyle, J.F. Dellaria, B.J.Siegfried, J. Org. Chem., 42, 1977, 2426-2430.

Intermediate 21

(3S,3aR,6aS)-1-Benzothiazol-2-yl-3-methyl-hexahydro-pyrrolo[3,2-b1pyrrol-2-one trifluoroacetate

To the Intermediate 20, (850mg, 2.194mmol) was added trifluoroacetic acid (20mL) at room temperature. After 20 minutes the solvent was evaporated in vacuo to give, after trituration under diethyl ether, the title compound as a tan solid, (643 mg). Mass spec 274 [MH]⁺.

Intermediate 22

(3aS,6S,6aR)-6-Methyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-sulphonic acid isopropyl-methyl-amide

Intermediate 18 ((3S,3aR,6aS)-3-Methyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one trifluoroacetate (438 mg, 1.723 mmol)) was stirred in anhydrous acetonitrile (5 mL) and treated with DIPEA (490 mg, 3.791 mmol, 660 μL) followed by N- isopropyl-N-methylsulphamoyl chloride¹ (325 mg, 1.893 mmol). The mixture was stirred at room temperature overnight; the solvent was removed in vacuo and the residue was shaken with ethyl acetate (100 mL) and water (20 mL). The organic phase was separated, washed with water, dried over anhydrous magnesium sulphate, filtered and evaporated in vacuo to give the title compound as a tan solid 300 mg (63%):

¹H nmr (CDCI₃): δ 6.2 (1H, broad s),4.14 (1H, septet), 3.82-3.70 (1H, m), 3.68- 3.35 (4H, m), 2.90-2.72 (1H, m), 2.76 (3H, s), 2.25-2.15 (1H, m), 1.30-1.14 (9H, m);

Mass spec, thermospray: +ve m/e: 276 [MH]⁺

T.J. Cheeseright, A.J. Edwards, D.T. Elmore, J.H. Jones, M. Raissi, E.C. Lewis; J.Chem.Soc.Perkin Trans.l; 12; (1994), 1595-1600

Intermediate 23

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1-sulphonic acid methylamide

(3S,3aR,6aS)-1-Benzothiazol-2-yl-3-methyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one trifluoroacetate (intermediate 21) (1.000 g, 2.58 mmol) was taken up in dichloromethane (200 mL) and shaken with saturated brine saturated with sodium carbonate (50 mL). The organic phase was separated, dried over anhydrous sodium sulphate, and evaporated under reduced pressure to give a red-brown oil. This was stirred with acetonitrile (10 mL) and DIPEA (494 μL, 1.1 equiv.) then N-methylsulphamoyl chloride¹ (500 mg, 1.5 equiv.) was added. The mixture was stirred at room temperature overnight, then taken up in ethyl acetate (300 mL) and shaken with 0.1 M hydrochloric acid (100 mL). The organic phase was separated, washed with water, dried over anhydrous magnesium sulphate, filtered and evaporated in vacuo to give a brown oil; this was purified by column chromatography on silica (Merck 9385) using 1 :2 ethyl acetate:cyclohexane as eluent to give the title compound (Rf 0.48 in 1:1 ethyl acetate:cyclohexane) as a colourless solid (420 mg, 44.5%) after trituration under dichloromethane/diethyl ether.

Η nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1H, dd), 7.31 (1H, dd), 4.30-4.17 (2H, m), 3.97-3.86 (1H, m), 3.76-3.62 (2H, m), 3.25-3.04 (2H, m), 2.86 (3H, d), 2.43- 2.26 (1H, m), 1.34 (3H, d):

Mass spec, thermospray: +ve m/e: 367 [MH]⁺

Ref 1. N-methyl-sulphamoyl chloride was prepared from methylamine hydrochloride by the method of W L Matier et al., J.Med.Chem.; 15; 1972; 538-541

Intermediate 24

[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1 -sulphonyQ-methyl-aminol-acetic acid

The intermediate f-butyl ester of example 24 (230 mg, 0.055 mmol) was dissolved in dichloromethane (2 mL) and treated with TFA (5 mL). The mixture was left at room temperature for thirty minutes then the solvents were evaporated in vacuo . The residue was triturated under diethyl ether to give the title compound (R, 0.6 in ethyl acetate:cyclohexane 1 :1) 200 mg (98%): ¹H nmr (CDCI₃): δ 7.84-7.76 (2H, m), 7.47-7.39 (1H. m), 7.35-7.27 (1H, m), 4.30- 4.04 (3H, m), 3.91-3.55 (3H, m), 3.25-3.00 (5H, m), 2.40-2.24 (1H, m), 1.35 (3H, d):

Mass spec, thermospray: +ve m/e: 457 [MH]⁺

Intermediate 25

3S,3aR,6aS)-3-Methyl-4-(methyl-propyl-sulphamoyl)-2-oxo-hexahydro- pyrrolo[3,2-b]pyrrole-1 -carboxylic acid tert-butyl ester

Intermediate 17 (277 mg, 1 mmol) was stirred in dichloromethane (5 mL) and N- methyl-N-propylsulphamoyl chloride (189 mg, 1.1 equiv.) was added. The stirred suspension was then treated with DIPEA (284.4 mg, 2.2 equiv., 383 μL) in one portion to give a homogeneous solution which was left at room temperature overnight. The mixture was partitioned between ethyl acetate and water (100 mL each). The organic phase was separated, dried over anhydrous magnesium sulphate and evaporated under reduced pressure to give a yellow oil, which was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1:1 ethyl acetate:cyclohexane as eluent to give the title compound (R_f 0.41 ) 330 mg (87.8%):

¹H nmr (CDCI₃): δ 3.83-3.70 (2H, m), 3.50-3.38 (2H, m), 3.29-3.05 (2H, m), 3.04- 2.93 (1H, m), 2.75 (3H, s), 2.52-2.45 (1H, m), 2.20-2.01 (1H, m), 1.70-1.50 (11H, m), 1.35 (3H, d), 0.93 (3H, t) Mass spec, thermospray: +ve m/e: 376 [MH]⁺

Intermediate 26

(3aS,6S,6aR)-6-Methyl-5-oxo-hexahydro-pyrrolo[3,2-blpyrrole-1-sulphonic acid methyl-propyl-amide

The intermediate 25 (280 mg) was dissolved in dichloromethane (2 mL) and treated with TFA (2 mL) (gas evolution was immediate). After 1 hour the solvents were removed under reduced pressure and the residue was triturated under cyclohexane to give the title compound (R_f 0,41 ) 195 mg (95%):

Η nmr (CDCI₃): δ 6.40 (1H, broad s), 3.83-3.74 (1H, m), 3.68-3.48 (3H, m), 3.29-

3.05 (2H, m), 2.90-2.75 (4H, m), 2.28-2.15 (1 H, m), 2.06-1.87 (1 H, m), 1.70-1.55

(2H, m), 1.35 (3H, d), 0.93 (3H, t)

Mass spec, thermospray: +ve m/e: 276 [MH]⁺

Intermediate 27 (3aS,6S,6aR)-4-[5-({[tert-butyl(diphenyl)silyl]oxy} methyl)-1 ,3-thiazol-2-yl]-N,6- dimethyl-5-oxo-N-propylhexahydropyrrolo[3,2-b] pyrrole-1 (2H)-sulphonamide

Intermediate 26 (1.156g, 4.2mmol) was dissolved in p-xylene (160ml) and treated with copper (ii) chloride (420mg, 4.24mmol), potassium carbonate (580mg, 4.2mmol), TDA-1 (453mg, 1.4mmol, 448ul), and intermediate 53

(2.17g, 5mmol). The mixture was heated at reflux using a Dean-Stark trap for 7h. The reaction was left to stand at room temperature overnight and then heated at reflux for a further 7h. After cooling to room temperature ethyl acetate (150ml) was added and the mixture filtered. The filtrate was washed with dilute aqueous hydrochloric acid solution (1N, 100ml), saturated sodium bicarbonate solution (100ml), dried (MgS04) and concentrated to give a brown gum. The crude product was purified by chromatography on silica (Merck 7729) using ethyl acetate yclohexane 5:95, 10:90 then 20:80 as eluent to give the title compound (Rf 0.3 in 20:80 ethyl acetate:cyclohexane) as a colourless gum (1.45g). - Η nmr (CDCI₃): δ 7.7-7.66 (4H,m), 7.46-7.36 (6H,m), 7.12 (1 H,s), 4.8 (2H,s), 4.05 (1 H,m), 3.82 (1 H,t), 3.67 (1 H,dd), 3.49 (1 H,m), 3.26-3.09 (3H,m), 2.87 (3H,s), 2.9-2.83 (1H,m), 2.25 (1H,m), 1.63 (2H,m), 1.28 (3H,d), 1.05 (9H,s), 0.93 (3H,t) LC/MS (method B): +ve m/e: 627 [MH]⁺ , RT = 4.49min

Intermediate 28

(3aS,6S,6aR)-6-Methyl-5-oxo-hexahydro-pyrrolo[3,2-blpyrrole hydrochloride Translactam intermediate 10 (5.865g, 21.38 mmol) was dissolved in IPA (310 mL). The solution was added to the catalyst (1.65 g, 10% palladium on activated charcoal with 50% water, Degussa type E101 NE/W) under nitrogen. The mixture was stirred vigorously under an atmosphere of hydrogen (copper sulphate hydrogenator). After a total of 3 hours 35 minutes the catalyst was removed by filtration through glass fibre filters and washed with a small amount of IPA. The filtrate and washings were combined and treated with 1 M hydrogen chloride in diethyl ether (22.5 mL, 22.5 mmol, 1.05 equiv). The mixture was evaporated under reduced pressure and dried in vacuo at room temperature overnight to give the title compound as a white foam (quantitative yield). MS (thermospray) 141 : [MH]⁺ and 281 [2M+H]⁺. H nmr (MeOD): δ 4.72 (3H + H₂0, s), 3.80 (1H, m), 3.60 (2H, m), 3.36 (1 H, dd), 2.69 (1 H, m), 2.27 (1 H, m), 1.82 (1 H, m), 1.14 (3H, d).

Intermediate 29 tert-butyl f[((3aS,6S,6aR)-6-methyl-5-oxohexahydropyrrolo[3,2-b1pyrrol-1(2H)- yl)sulphonyl](methyl)amino]acetate

Intermediate 28 (3S,3aR,6aS)-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one hydrochloride (1.08g, 6.09mmol) was suspended in dry acetonitrile (10ml), di- isopropylethylamine (2.3ml, 0.013mmol) then tert-butyl [(chlorosulphonyl) (methyl)amino]acetate (intermediate 41) (1.69g, 6.9mmol) were added and dichloromethane (2ml) and di-isopropylethylamine (1 ml, 5.74mmol) were added to dissolve any remaining solids, the mixture was left under N₂ for 3 days. Ethyl acetate (40ml) was added washed with 0.01 M aqueous hydrochloric acid (50ml) then brine (25ml). The aqueous phase was re-extracted twice with ethyl acetate (75ml). The combined organics were dried over anhydrous magnesium sulphate, filtered and evaporated under reduced pressure to yield a golden oil. This was taken up in dichloromethane and purified by flash column chromatography (Merck 9385), eluted with 4:1 ethyl acetate:cyclohexane to give the title compound as a white gum 404mg (33%). ¹H NMR (400MHz, CDCI₃) δ 1.22 (3H, d), 1.48 9H, s), 1.91-2.03 (1H, m), 2.13-2.24 (1H, m), 2.78-2.86 (1H, m), 3.02 (3H, s), 3.52-3.63 (3H, m), 3.73-3.82 (1 H, m), 3.92 (2H, AB quartet), 5.88 (1H, brd s) Mass Spectrum (thermospray, +ve ion) m/e = 348 (M+H)⁺

Intermediate 30 tert-butyl [[((3aS,6S,6aR)-4-[5-({[tert-butyl(diphenyl)silyl]oxy}methyl)-1 ,3-thiazol- 2-yl]-6-methyl-5-oxohexahydropyrrolof3,2-blpyrrol-1(2H)- yl)sulphonyl1(methyl)amino]acetate

intermediate 29 tert-butyl [[((3aS,6S,6aR)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]acetate (404mg, 1.16mmol) was dissolved in p-xylene (40ml) and potassium carbonate (240mg, 1.74mmol), 2- bromo-5-({[tert-butyl(diphenyl)silyl]oxy}methyl)-1 ,3-thiazole (intermediate 53) (752mg, 1.74mmol), TDA-1 (112μl, 0.35mmol) and copper (I) chloride (115mg, 1.16mmol) were added. The mixture was stirred at reflux under N₂ for 7.5 hours then left at room temperature for 19 hours and refluxed again for 3 hours. The mixture was cooled, diluted with ethyl acetate (60ml), washed with 0.2M aqueous hydrochloric acid (100ml) and saturated aqueous sodium chloride (20ml). The organic layer was re-extracted twice with ethyl acetate (100ml). The organic extracts were dried over magnesium sulphate, filtered, evaporated under reduced pressure and dried in vacuo to yield a brown gum. This was taken up in dichloromethane and purified by flash column chromatography, silica gel Merck 9385, eluent 10:1 cyclohexane:ethyl acetate, to give the title compound as a white gum 374mg (46%): ¹H NMR (250MHz, CDCy δ 1.06 (9H, s), 1.30 (3H, d), 1.49 (9H, s), 2.18-2.38 (1 H, m), 2.88-2.93 (1H, m), 3.03 (3H, s), 3.06-3.20 (1 H, m), 3.60-4.14 (4H, m), 4.82 (2H,s), 7.13 (1 H, s), 7.34-7.49 (4H, m), 7.66-7.73 (6H, m) LC-MS (A, 5.5min) Peak at 4.44min gives m/e 699 (M+H)⁺

Intermediate 31 tert-butyl [[((3aS,6S,6aR)-4-[5-({[(ethylamino) carbonyl]oxy}methyl)-1 ,3-thiazol-2- yl]-6-methyl-5-oxohexahydropyrrolo[3,2-b] pyrrol-1 (2H)- yl)sulphonyl](methyl)amino]acetate Example 78 (115mg, 0.25mmol) was dissolved in dichloromethane (11ml), triethylamine (34.8μl, 0.26mmol) then ethyl isocyanate (20.6μl, 0.26mmol) were added. The mixture was left at room temperature for 7 days, further amount of ethyl isocyanate (1 OOμl, 1.26mmol) was added, then after 3 days further amount of ethyl isocyanate (100μl, 1.26mmol) was added and after the 2 days the remaining yellow gum was dissolved in dichloromethane and purified by solid phase extraction, Bond-Elut silica column 1g, eluted with ethyl acetate cyclohexane mixtures to give the title compound as a white foam, 94.3mg (71%): ¹H NMR (250MHz, CDCI₃) δ 1.14 (3H, t), 1.28 (3H, d), 1.48 (9H, s), 2.26 (1 H, m), 2.86 (1 H, m), 3.03 (3H, s),

3.06-3.38 (3H, m), 3.60-4.14 (7H, m), 4.62 (2H,s), 7.37 (1 H, s)

LC-MS (A, 5.5min)

Peak at time 3.36 minutes gives m/e 532 (M+H)⁺

Intermediate 32 rr((3aS,6S,6aR)-4-r5-({[(ethylamino)carbonyl] oxy}methyl)-1 ,3-thiazol-2-yl]-6- methyl-5-oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)- yl)sulphonyl](methyl)amino]acetic acid

Intermediate 31 (21 mg, 0.0395mmol) was dissolved in dichloromethane (500μl).

Trifluoroacetic acid (500μl, 6.49mmol) was added and the mixture was stirred for

35 minutes at room temperature after which dichloromethane (500μl) and toluene (1 ml) were added and the mixture was evaporated under reduced pressure and dried in vacuo to give the title compound as a white solid, 17.4mg

(93%):

¹H NMR (400MHz, CDCI₃) δ 1.10 (3H, t), 1.28 (3H, d), 2.25 (1 H, m), 2.85 (1 H, m), 3.05 (3H, s), 3.08-3.28 (3H, m), 3.60-3.87 (1 H, m), 3.87-4.02 (2H, m), 4.30 (3H, brd s), 4.74 (1 H, s),

5.17-5.28 (2H, m), 7.39 (1 H, s)

LC-MS (A, 5.5min)

Peak at 2.80min gives m/e 476 (M+H)⁺ and 474 (M-H)^" Intermediate 33

(3aS,6S,6aR)-4-[4-({[tert-butyl(diphenyl)silyl]oxy}methyl)-1 ,3-thiazol-2-yl]-N,6- dimethyl-5-oxo-N-propylhexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

Intermediate 26 (0.578g, 2.1 mmol), intermediate 54 (1.09g, 2.52mmol), TDA-1 (0.227g, 0.224ml, 2.12mmol), potassium carbonate (0.290g, 2.1 mmol) and copper (I) chloride (0.21g, 2.12mmol) were suspended in p-xylene (80ml). The reaction mixture was heated at reflux with a Dean and Stark trap for 5 hours, left to stand for 15 hours and then refluxed for a further 6 hours. The reaction mixture was allowed to cool, diluted with ethyl acetate and filtered. The filtrate was washed with 2N hydrochloric acid, saturated sodium bicarbonate solution and brine. The organic phase was dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (6:1 v/v) as eluant to give the title compound as a colourless gum, 0.463g (35%).

¹H NMR (CDCI₃): δ 7.70 (4H, m), 7.39 (6H, m), 6.93 (1 H, t, J=1 Hz), 4.76 (2H, d, J=1 Hz), 4.01 (1 H, m), 3.79 (1 H, m), 3.65 (1 H, dd, J=6, 10Hz), 3.44 (1 H, m), 3.29-3.07 (3H, m), 2.87 (3H, s), 2.78 (1H, m), 2.17 (1H, m), 1.63 (2H, hextet, J=7Hz), 1.28 (3H, d, J=7.5Hz), 1.10 (9H, s), 0.93 (3H, t, J=7Hz). Mass spec, thermospray: +ve m/e: 627 [MH⁺]

Intermediate 34

(3aS,6S,6aR)-4-[4-(2-{[tert-butyl(diphenyl)silyl]oxy}ethyl)-1 ,3-thiazol-2-yl]-N,6- dimethyl-5-oxo-N-propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Intermediate 26 (0.578g, 2.1mmol), intermediate 56 (1.12g, 2.52mmol), TDA-1 (0.227g, 0.224ml, 2.12mmol), potassium carbonate (0.290g, 2.1 mmol) and copper (I) chloride (0.21g, 2.12mmol) were suspended in p-xylene (80ml). The reaction mixture was heated at reflux with a Dean and Stark trap for 6 hours.

The reaction mixture was allowed to cool, diluted with ethyl acetate and filtered. The filtrate was washed with 2N hydrochloric acid, saturated sodium bicarbonate solution and brine. The organic phase was dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~-4psi) and cyclohexane/ethyl acetate (6:1 v/v) as eluant to give the title compound as a colourless gum, 0.40g

(30%).

Η NMR (CDCI₃): δ 7.58 (4H, m), 7.37 (6H, m), 6.64 (1 H, s), 4.04-3.91 (3H, m),

3.77 (1 H, m), 3.62 (1 H, dd, J=6, 10Hz), 3.43 (1 H, m), 3.30-3.06 (3H, m), 2.90

(2H, t, J=6Hz), 2.88 (3H, s), 2.69 (1 H, m), 2.07 (1 H, m), 1.63 (2H, hextet,

J=7Hz), 1.27 (3H, d, J=7.5Hz), 1.02 (9H, s), 0.94 (3H, t, J=7Hz).

Mass spec, thermospray: +ve m/e: 641 [MH⁺]

Intermediate 35

(3aS,6S,6aR)-4-[6-({[tert-butyl(diphenyl)silyi]oxy} methyl)-1 ,3-benzothiazol-2-yl]- N,6-dimethyl-5-oxo-N-propylhexahydropyrrolo [3,2-b]pyrrole-1(2H)- sulphonamide

The product was similarly prepared to intermediate 44 by reacting intermediate 26 with intermediate 57 and was obtained as a cream foam in 42% yield. ¹H nmr (CDCI₃) : δ 7.79-7.64 (6H,m,arH), 7.69 (1 H, br.s.NH), 7.48-7.31 (7H,m, arH), 4.85 (2H,s, arCH₂), 4.27-4.11 (1 H,m,5-H), 3.88 (1H,t,6a-H), 3.74 (1H,dd,5- H or 3a-H), 3.63-3.48 (1 H, m, 5-H or 3a-H), 3.33-2.98 (4H,m,3-H,6-H,CH₂N ), 2.90 (3H,s,N-CH₃), 2.44-2.23(1 H,m,6-H), 1.73- .56(2H,m,MeCH₂), 1.32 (3H,d,CH₃), 1.10 (9H,s, t-Bu), 0.95(3H,t,CH₃) ppm. Mass Spec. : 677 [MH]⁺

Intermediate 36 tert-butyl [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-benzothiazol-6-yl]methyl{[2-

(trimethylsilyl)ethyl]sulphonyl}carbamate

To example 94, (61 mg,139μmol, 1eq.) in dry THF(5 mL),stirred at room temperature under nitrogen, was added ter.-butyl[[2-[(trimethylsilyl)ethyl] sulphonyl] carbamate¹ (82 mg, 293μmol2.1eq.) and triphenylphosphine (79.4mg,303μmol, 2.2eq.). The solution was cooled (ice/water bath) and Diethylazodicarboxylate (33μL,209μmol, 1.5eq.) was added over ca. 5 min.. The reaction mixture was allowed to stir at room temperature for 4.5 hours. The solvent was evaporated in vacuo and the product was purified by flash column chromatography over silica (Merck 9385 silica gel, 60 mL), eluting with cyclohexane:ethyl acetate (8:2). The title compound was obtained as a white foam (101.8mg, 104%). ¹H nmr (CDCI₃) : δ 7.85 (1 H,s,arH), 7.74 (1 H, d, arH), 7.50 (1 H,d, arH), 4.95 (2H, s,arCH₂), 4.27-4.08 (1H,m,5-H), 3.88 (1H,t,6a-H), 3.75 (1H,dd,5-H or 3a-H), 3.63-3.50 (1 H,m,5-H or 3a-H), 3.42-2.99 (6H,m,6-H,3-H,CH₂N,CH₂S0₂ ), 2.91 (3H,s,N-Me), 2.44-2.23 (1H,m,6-H), 1.78-1.58 (2H,m, CH₂), 1.51 (d,t-Bu), 1.33- (3H,d,CH3), 1.11-0.76 (5H,m,CH₃, CH₂Si), 0.0 (9H,d, Si (Me)₃) ppm. Mass Spec. : 702 [MH]⁺, 299, 199. HPLC: 39.1 min. (100%). LCMS: 4.11 min. 702 [MH]⁺.

Ref 1 Jeffrey A. Campbell, David J. Hart, J. Org. Chem. 1993, 58, 2900-2903. Intermediate 37 N-methyl-N-propylsulphamoyl chloride

N-methylpropylamine (5.000 g, 68.36 mmol) in dry DCM (30 mL) was added dropwise over 30 mins to a stirred solution of sulphuryl chloride (10.96 mL, 2 equiv.) in dry DCM (50 mL) at -10°C with exclusion of moisture. After the mixture had been stirred at -10°C for a further 1 hr, it was evaporated under reduced pressure to give a yellow oil. This was taken up in ethyl acetate (100 mL) and washed with water (50 mL). The organic phase was separated, dried over MgS04, filtered and evaporated under reduced pressure to give the title compound as a colourless liquid (4.82 g, 41%): ¹H nmr (CDCI₃250 MHz) : δ 3.25 (2H, t, J 7Hz), 3.00 (3H, s), 1.70-1.65 (2H, m), 1.00 (3H, t, J 7Hz)

Intermediate 38

Similarly prepared to intermediate 37 N-methyl-N-isopropylsulphamoyl chloride colourless liquid, 16%):

Η nmr (CDCI₃, 250 MHz) : δ 4.49-4.33 (1 H, septet, J 7Hz), 2.93 (3H, s), 1.27 (6H, d, J 7Hz) Intermediate 39

Similarly prepared to intermediate 37 N-methyl-N-isobutylsulphamoyl chloride colourless liquid, 26%): Η nmr (CDCI₃, 250 MHz) : δ 3.06-2.94 (5H, m ), 1.95 (1 H, m), 1.00 (6H, d, J 7Hz)

Intermediate 40 N-methyl-N-allylsulphamoyl chloride

N-methylallylamine (5.000 g, 70.31 mmol) and triethylamine (7.101 g, 9.78 mL, 1 equiv.) in 40-60° petroleum ether (140 mL) were added dropwise to a stirred solution of sulphuryl chloride (9.489 g, 5.7 mL, 1 equiv.) in 40-60° petroleum ether (350 mL) at -78oC under nitrogen. After addition was complete, the cooling-bath was removed and the mixture was allowed to warm to room temperature then stirred at room temperature for 1 hr. The triethylamine hydrochloride was removed by filtration, and the solid was washed with 40-60° petroleum ether (2x200 mL). The combined filtrate was evaporated under reduced pressure to give the title compound as a colourless oil (9.6 g, 80%): ¹H nmr (CDCI₃, 400 MHz) : δ 5.93-5.83 (1 H, m ), 5.42-5.39 (1 H, m), 5.37-5.35 (1 H, m), 3.93-3.83 (2H, broad m), 2.93 (3H, s)

Intermediate 41 tert-butyl [(chlorosulphonyl)(methyl)amino]acetate

Sarcosine t-butyl ester¹ (4.4g, 30.3mmol) and triethylamine (4.21 mmol, 30.3mmol) were dissolved in dichloromethane (140ml). This was cautiously added to sulphuryl chloride (4.9ml, 60.4mmol) in dichloromethane (45ml), maintaining te temperature below -20°C, under N₂. The mixture was allowed to stir at room temperature for 1 h, then evaporated to give a greenish-white solid. This was dissolved in dichloromethane (100ml) and washed with cold water (100ml). The organic layer was separated, dried over magnesium sulphate, filtered and evaporated to give the title compound as a yellow oil 4.34g, 59%. ¹H NMR (400MHz, CDCI₃) 1.5 (s, 9H), 3.1 (s, 3H), 3.95 (s, 2H)

Ref 1 C. Florine, M. Rolland, J. Verducci; Organic Preparations Proceedings International 26(5);

(1994), 608-610

Intermediate 42

Benzyl [(chlorosulphonyl)(methyl)amino]acetate

Sarcosine hydrochloride (10.78g, 50mmol) was suspended in DCM (200ml) and to this was added triethylamine (15ml, 107.5mmol). This mixture was added dropwise to a solution of sulphuryl chloride (9.25ml, lOOmmol) in DCM (75ml) at 0°C under nitrogen. After 1.5 hours water (150ml) was added to the reaction and the organic phase was separated. The organic phase was washed with brine, dried (anhydrous magnesium sulphate) and evaporated in vacuo. The residue was chromatographed over silica gel using cyclohexane/ethyl acetate (2:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a pale yellow oil, 5.57g (40%). ¹H nmr (CDCI₃): δ 7.38 (5H, m), 5.22 (2H, s), 4.09 (2H, s), 3.13 (3H, s).

Intermediate 43

(3aS,6S,6aR)-N-allyl-N,6-dimethyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1(2H)- sulphonamide

Intermediate 17 (2.77 g, 10 mmol) was stirred in acetonitrile (20 mL) and DIPEA (2.844 g, 2.2 equiv., 3.833 mL) was added, followed by N-methyl-N-allylsulphamoyl chloride (intermediate 40) (1.696 g, 1 equiv., prepared from N-methyl-N-allylamine by the procedure of Anseime¹ ). The mixture was stirred at room temperature overnight then partitioned between ethyl acetate and 0.01 M hydrochloric acid aq. The organic phase was separated, dried over anhydrous magnesium sulphate, filtered and evaporated under reduced pressure to give a yellow gum. This was purified by flash chromatography on silica (1 :9 ethyl acetate yclohexane) and the purified product was dissolved in dichloromethane (10 mL) and treated with TFA (10 mL). After one hour, the TFA and DCM were removed under reduced pressure to give a tan gum which was purified by flash chromatography on silica (1:1 ethyl acetate:cyclohexane) to give the title compound as a colourless solid Rf 0.35 in ethyl acetate (76% yield over the two steps):

¹H nmr (CDCI₃) (400 MHz): δ 7.19 (1H, broad s)), 5.86-5.76 (1H, m), 5.32-5.24

(2H, m), 3.89-3.72 (3H, m), 3.68-3.63 (1H, m), 3.55-3.43 (2H, m), 2.90-2.78 (4H, m), 2.27-2.20 (1H, m), 2.24-1.94 (1 H, m), 1.23 (3H, d):

Mass spec, thermospray: +ve m/e: 274 [MH]⁺

Ref. 1 J-P Anseime et al., Bull. Soc. Chim. Belg. 93 10) 919 (1984)

Intermediate 44

(3aS,6S,6aR)-N-allyl-4-[5-({[tert-butyl(diphenyl) silyl]oxy}methyl)-1 ,3-thiazol-2-yl]- N,6-dimethyl-5-oxohexahydropyrrolo[3,2-b] pyrrole-1(2H)-sulphonamide

A mixture of intermediate 43, (279mg, 1.02mmol, 1eq), copper (1) chloride (102mg, 1.03mmol, 1eq), potassium carbonate (163mg, 1.18mmol, 1.15eq), TDA-1 (98.1 μL, 0.31 mmol, 0.3eq) and intermediate 53, 545mg, 1.26mmol, 1.23eq) in p-xylene (20mL) was stirred and refluxed under nitrogen. The oil bath temperature was ca. 160°C. The mixture was stirred at reflux for 5 hr 20 min, and then allowed to cool to room temperature. Then the mixture was diluted with ethyl acetate (20mL) and washed with 0.2M hydrochloric acid (20mL). The aqueous phase was extracted with ethyl acetate (20mL). The combined organic phase was dried (MgS0₄) and evaporated to leave a dark brown gum. Purification by flash column chromatography over silica gel ( Merck 9385) using cyclohexane:ethyl acetate (7:1) as the eluting solvent system gave 349mg (54%) of title compound as a white foam. ¹H nmr (CDCI₃): δ 7.69 (4H, m), 7.41 (6H, m), 7.13 (1H, s), 5.82 (1H, m), 5.31 (1H, m), 5.25 (1H, m), 4.81 (2H, s), 4.09 (1H, m), 3.86 (2H, m), 3.80 (1H, m), 3.70 (1H, dd), 3.54 (1H, m), 3.15 (1H, m), 2.89 (1H, m), 2.86 (3H, s), 2.27 (1H, m), 1.30 ( 3H, d), 1.06 (9H, s). Mass spec 625 [MH]⁺, (thermospray).

Intermediate 45 benzyl [[((3aS,6S,6aR)-4-{f(2R,3S)-2,3-dimethylcyclopropyl]carbonyl)-6-methyl- 5-oxohexahydropyrrolof3,2-b1pyrrol-1(2H)-yl)sulfonyl](methyl)aminolacetate To a solution of intermediate 16 (3.28g, 12mmol) and DIPEA (6.1ml, 36mmol) in DCM (75ml) was added a solution of intermediate 42 (5.0g, 18mmoi) in DCM (25ml) and the reaction mixture was stirred at room temperature under nitrogen for 3 hours. The reaction mixture was washed with 2N hydrochloric acid, water, saturated sodium bicarbonate solution and brine. The organic phase was dried (anhydrous magnesium sulphate) and evaporated in vacuo. The residue was chromatographed over silica gel using cyclohexane/ethyl acetate (4:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a pale yellow oil, 4.53g (79%).

¹H nmr (CDCI₃): δ 7.37 (5H, m), 5.19 (2H, s), 4.05 (2H, s), 3.67 (2H, m), 3.55- 3.43 (2H, m), 3.01 (3H, s), 2.97 (1H, m), 2.82 (1H, t, J=9Hz), 2.62 (1H, m), 1.96 (1H, m), 1.70-1.53 (2H, m), 1.23-1.16 (9H, m). LC-MS (method A): Peak at 3.74 mins gives m/e 478 [MH⁺]

Intermediate 46

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5- oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

Intermediate 16 (1.02g, 3.74mmoi) was dissolved in dry acetonitrile (20ml). N- methylsulphamoyl chloride¹, 951mg, 7.34mmol) in dry DCM (10ml) was added and the mixture stirred under N₂ for 5 minutes. DIPEA (2ml, 11.5mmol) was slowly added over 10 minutes. The mixture was stirred for a further 2 hours and left to stand overnight. The solvent was evaporated under educed pressure and the residue taken up in ethyl acetate (40ml), washed with water (20ml) and the aqueous phase re-extracted with ethyl acetate (40ml). The combined organics were dried over magnesium sulphate, filtered and evaporated to give an orange- white solid (1.74g). This was taken up in DCM and purified by flash column chromatography, silica gel 9385, eluent 4:1 cyclohexane:ethyl acetate, to give the title compound, white solid, 1.04g (86%). ¹H NMR (400MHz, CDCI₃) δ 1.15-1.21 (6H, m), 1.28 (3H, d), 1.54-1.69 (2H, m), 1.95-2.07 (1 H, m), 2.66-

2.75 (1 H, m), 2.82-2.07 (4H, m), 2.96-3.05 (1 H, m), 3.42-3.57 (2H, m), 3.74-3.86

(2H, m), 4.11-4.17 (1H,m)

LC-MS (A)

Peak at time 3.09min gives m/e 330 (M+H)⁺

Ref 1. N-methyl-sulphamoyl chloride was prepared from methylamine hydrochloride by the method of W L Matier et al., J.Med.Chem.; 15; 1972; 538-541

Intermediate 47

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonyl chloride

Intermediate 16 (2.18g, 8.0 mmol) was dissolved in dry dichloromethane (100 mL) at room temperature, under nitrogen. Triethylamine (2.8 mL, 20.09 mmol, 2.5 equiv) was added to the stirred mixture followed by chlorotrimethylsilane (1.45 mL, 11.42 mmol, 1.42 equiv). The mixture was stirred for 3 hours before sulphuryl chloride (1.3 mL, 11.22 mmol, 1.4 equiv) was added. The mixture was stirred a further 2 hours before the solvent was removed by evaporation under reduced pressure (cold water bath). The residue was partitioned between dichloromethane (75 mL) and water (30 mL). The organic phase was washed with saturated brine (25 mL), dried (MgS0₄) and evaporated to leave the title compound as a grey foam, (2.274g, 6.79 mmol, 85%). ¹H nmr (CDCI₃) (400 MHz): δ 3.96-3.80 (3H, m), 3.46-3.40 (1H, m), 3.08-3.00 (1H, m), 2.86-2.28 (2H, m), 2.14-2.00 (1 H, m), 1.73-1.57 (2H, d), 1.30 (3H, d), 1.22-1.15 (6H, m): LC/MS (A, 5.5min) Peak at 3.52 min gives m/e 358 [MNa]⁺and 691 [2M+Na]⁺.

Intermediate 48

(3aS,6S,6aR)-N-isopropyl-N,6-dimethyl-5-oxohexahydropyrrolo [3,2-b]pyrrole- 1 (2H)-sulphonamide Intermediate 18 (483 mg, 1.723 mmol) was stirred at room temperature in dry acetonitrile (5 mL) and treated with DIPEA (660 μL, 2.2 equiv.), followed by N- isopropyl-N-methylsulphamoyl chloride (prepared by the method of Jones et al.¹) (325 mg, 1.1 equiv.). The mixture was stirred at room temperature overnight, then evaporated in vacuo and the residue was partitioned between water and ethyl acetate. The organic phase was separated, dried (MgS04), and evaporated in vacuo to give the title compound as a tan solid (300 mg, 63%) after trituration under diethyl ether:

¹H nmr (CDCI₃) (250 MHz): δ 6.70 (1H, broad s), 4.21-4.07(1 H, m), 3.81-3.69 (1 H, m), 3.66-3.35 (3H, m), 2.90-2.73 (4H, m), 2.26-2.17 (1H, m), 2.05-1.86 (1 H, m), 1.30-1.15 (9H, m) Mass spec, thermospray: +ve m/e: 276 [MH]⁺

Ref. 1 T . Cheeseright, A.J. Edwards, D.T. Elmore, J.H. Jones, M. Raissi, E.C. Lewis; J.Chem.Soc.Perkin Trans.1; 12; (1994), 1595-1600

Intermediate 49

(1R,6S)-bicyclo[4.1.0]hept-3-ene-7-carboxyiic acid was prepared by the method of H Musso and U. Bietham , Chem Ber, 1964, 97, 2282

Intermediate 50

(1R,5S)-bicyclo[4.1.0]hex-2-ene-6-carboxylic acid was prepared by the method of J. Meinwald, S.S. Labana and M.S. Chadha J. Am. Chem. Soc, 85, 582-585 (1963)

Intermediate 51 2-bromo-4-formylthiazole

2-Amino-4-formylthiazole¹ (1.0 g, 7.81 mmol) was slurried with acetonitrile (15ml) and added portionwise to a stirred mixture of copper (II) bromide (2.09g, 9.37mmol) and t-butyl nitrite (1.4ml, 11.71 mmol) in acetonitrile (60ml) at room temperature. The reaction mixture was left to stir overnight and then treated with 2N sodium hydroxide solution (120ml). The mixture was extracted with ethyl acetate and the organic extracts dried (MgS0₄) and concentrated. The crude product was purified by chromatography on silica (Merck 9385) using ethyl acetate yclohexane 1:1 as eluent to give the title compound (Rf 0.7 in 1:1 ethyl acetate:cyclohexane) as a white solid (0.84g).

Η nmr (CDCI₃): δ 10.0 (1H,s), 8.2 (1 H,s)

Ref 1 P N Plazzi et al, Farmaco (1989), 44(11), 1011-30

Intermediate 52 2-bromo-4-hydroxymethylthiazole

Intermediate 51 (0.8g, 4.167mmol) was dissolved in methanol (12ml) and stirred at room temperature under nitrogen. Sodium borohydride (0.081g, 2.16mmol) was added portion wise over30 mins. After 2h the reaction mixture was concentrated and the residue chromatographed on silica (Merck 9385) eluting with 1:1 cyclohexane:ethyl acetate to give the title compound (Rf 0.25 in 1:1 ethyl acetate yclohexane) as a yellow oil (0.507g) ¹H nmr (CDCI₃): δ 7.45 (1H,s), 4.8 (2H,d), 2.1 (1H, t)

Intermediate 53

2-bromo-5-({[tert-butyl(diphenyl)silyl]oxy}methyl)-1 ,3-thiazole

Intermediate 52 (8.2g, 42.3mmol) was dissolved in DMF (50ml) and treated with imidazole (6.9g, 101.4mmo!) followed by tert-butyldiphenylsilyl chloride (13.9g, 50.7mmol). The reaction mixture was stirred at room temperature for 18h. Water (50ml) was added followed by dilute aqueous hydrochloric acid (1Ν, 100ml). The mixture was extracted with ether (2 x 100ml) and the combined organic extracts washed with water (100ml), aqueous hydrochloric acid (1N, 100ml), saturated sodium bicarbonate solution (100ml), and brine (100ml). The organic was dried (MgS0₄) and concentrated to give a yellow oil. . The crude product was purified by chromatography on silica (Merck 7729) using DCM yclohexane 1 :1 then 2:1 as eluent to give the title compound (Rf 0.24 in 1 :1 DCM:cyclohexane) as a white solid (15.65g). ¹H nmr (CDCI₃): δ 7.65 (4H,m), 7.4 (6H,m), 7.25 (1H, s), 4.8 (2H,s), 1.1 (9H,s) Mass spec, thermospray: +ve m/e: 432/434 [MH]⁺ Intermediate 54

(2-bromo-1 ,3-thiazol-4-yl)methyl tert-butyl(diphenyl)silyl ether

To a solution of 2-bromo-4-(hydroxymethyl)thiazole¹ (1.37g, 7.1 mmol) in DMF (50ml) was added imidazole (1.36g, 20mmol) and tBDPSCI (2.6ml, 2.75g, 10mmol). The reaction mixture was stirred under nitrogen at room temperature for 1.5 hours. The reaction mixture was separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with brine, dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (20:1 v/v) as eluant to give the title compound as a pale yellow oil, 2.658g (87%). ¹H NMR (CDCI₃): δ 7.67 (4H, s), 7.38 (6H, m), 7.21 (1 H, s), 4.85 (2H, s), 1.10 (9H, s).

Mass spec, thermospray: +ve m/e: 432, 434 (ratio 1 :1) [MH⁺] Ref 1 Benoit, Marc; Demoute, Jean Pierre; Wehrey, Christian. Preparation of thiazolylmethyl pyrethroid esters as pesticides. Eur. Pat. Appl., EP 556123 A1 930818.

Intermediate 55 2-(2-bromo-1,3-thiazol-4-yl)ethanol

To a mixture of 2-amino-4-(2-hydroxyethyl)thiazole¹ [2] (6.34g, 44mmol), copper sulphate pentahydrate (33g, 132mmol) and sodium bromide (18.05g, 176mmol) in 9M sulphuric acid (60ml), in an ice-salt bath at -5°C, was added a solution of sodium nitrite (3.64g, 52.8mmol) in water (5ml) over 30 minutes maintaining the temperature below 0°C. The reaction mixture was allowed to warm to room temperature over 1 hour and then diluted with water (60ml). This was extracted using ethyl acetate (x3) and the combined organics were washed with brine and dried over anhydrous magnesium sulphate. The aqueous phase was basified using 2N sodium hydroxide solution and extracted using ethyl acetate (x2). These extractions were combined, washed with brine and dried over anhydrous magnesium sulphate. The two organic phases were evaporated in vacuo, combined and chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (2:1 v/v) as eluant. This gave the title compound as an orange oil, 1.47g (16%).

Η NMR (D₆-DMSO): δ 7.38 (1 H, s), 4.68 (1 H, broad s), 3.67 (2H, t, J=7Hz), 2.82

(2H, t, J=7Hz).

Mass spec, thermospray: +ve m/e: 208, 210 (ratio 1 :1) [MH⁺]

Ref 1 Hartman, George D.; Duggan, Mark E.; Ihie. Nathan C; Hoffman, William F. N-

(Guanidinoalkoxybenzoyl)-α-(phenylsulphonylamino)-β-aianine derivatives and analogues for inhibiting osteoclast-mediated bone resorption. PCT Int. Appl., WO 9532710 A1 951207.

Intermediate 56

2-(2-bromo-1 ,3-thiazol-4-yl)ethyl tert-butyl(diphenyl)silyl ether

To a solution of intermediate 55 (1.50g, 7.2mmol) in DMF (100ml) was added imidazole (1.18g, 17mmol) and tBDPSCI (2.25ml, 2.37g, 8.6mmol). The reaction mixture was stirred under nitrogen at room temperature for 2 hours. The reaction mixture was separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with brine, dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (20:1 v/v) as eluant to give the title compound as a colourless oil, 2.745g (85%). ¹H NMR (CDCI₃): δ 7.58 (4H, s), 7.38 (6H, m), 6.95 (1H, s), 3.97 (2H, t, J=6Hz), 2.98 (2H, t, J=6Hz), 1.00 (9H, s).

Mass spec, thermospray: +ve m/e: 446, 448 (ratio 1 :1) [MH⁺]

Intermediate 57 2-Bromo-6-(tert-butyl-diphenyl-silanyloxymethyl)-benzothiazole was prepared by the method of intermediate 34 in WO 98/43975 (Borthwick et al, GlaxoGroup Limited patent application WO 98/43975)

Intermediate 58 (3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-4-[(3-chloropropyl)sulphonyl]-3- methylhexahydropyrroio[3,2-b]pyrrol-2(1 H)-one

Intermediate 21 (67.3 mg) in dry acetonitrile was treated with triethylamine (73 μL) and 3-chloropropanesulphonyl chloride (25 μL). The mixture was stirred at room temperature overnight, then quenched by addition of 3 drops of isopropanol and evaporated in vacuo. The residue was partitioned between water and DCM. The organic phase was separated, washed with aqueous sodium hydrogen carbonate and dried (MgS0₄). The crude material was purified by Bond-elut cartridge (eluting with cyclohexane, cyclohexane:diethyl ether (2:1 ) and cyclohexane:ethyl acetate (4:1 ) to give the title compound as a colourless solid (61 mg, 85%): LC-MS (method A): Peak at time 3.38 minutes gives m/e = 414/416 (MH)⁺

Intermediate 59

(3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-4-f(3-iodopropyl)sulphonyl]-3- methylhexahydropyrrolo[3,2-b]pyrrol-2(1H)-one

Intermediate 58 (57 mg) was treated with an excess of sodium iodide (15 equiv.) in acetone at reflux for 17 hours. The mixture was filtered, evaporated in vacuo, then partitioned between water and ethyl acetate. The organic phase was separated and dried (MgS0₄). Purification by Bond-elut cartridge gave the title compound as a colourless foam (57 mg, 82%): LC-MS (method A): Peak at time 3.51 minutes gives m/e = 506 (MH)⁺

Intermediate 60

(3aS,6S,6aR)-N-[4-(bromomethyl)benzyl]-4-{[(2R,3S)-2,3- dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5-oxohexahydro pyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide Intermediate 46 (120mg, 0.36mmol) and α,α'-dibromo-p-xyiene (386mg, 1.46mmol) was dissolved in acetonitrile:DMF (2:1 , 6ml) and treated with caesium carbonate (476mg, 1.46mmol). The mixture was stirred at room temperature for 3h. The solvents were removed in vacuo and the residue diluted with ethyl acetate (50ml) and filtered. The filtrate was washed with water, dried (MgS04) and concentrated to give a white solid. The crude product was purified on a 10g SPE silica cartridge eluting with DCM (x3), diethyl etheπcyclohexane (1:4,), diethyl etheπcyclohexane (1 :1 ), ether, and ethyl acetate to give the title compound (Rf 0.27 in 1 :4 ethyl acetate: cyclohexane) as a colourless gum (123mg)

LC/MS (method A)

Peak at 3.86 mins gives m/e 512/514 [MH⁺]

Intermediate 61 benzyl (3S)-2-methoxy-3-[(2,2,2-trifluoroacetyl)amino1pyrrolidine-1-carboxylate

benzyl (3S)-2-oxo-3-[(2,2,2-trifluoroacetyl)amino]pyrrolidine-1-carboxylate (prepared by the method of intermediate 56 in WO 98/43975 (Borthwick et al, GlaxoWellcome patent WO 98/43975)) (490g, 1.48mol) was dissolved in dry THF (2000ml) and LiBH₄ (800ml, 1.6mol, 2.0M solution in THF) was added under N₂, temperature < -20°C. After the addition was complete the mixture was stirred for 2 hours. In a separate vessel acetyl chloride (162ml, 2.27mol) was added to methanol (2000ml) maintaining temperature < 20°C. The solution of the reduced starting material was added to the acidic methanol, temperature < 25°C. NaHC0₃ (250g) in water (1200ml) was added with stirring and the mixture left to stand overnight.

NaCl (168g) in water (3000ml) and t-butyl methyl ether (1250ml) were added and the mixture separated. The aqueous phase was extracted with f-butyl methyl ether (1250ml). The combined organics were washed with brine (2000ml), dried over MgS0₄, filtered and evaporated to give a cloudy oil. This was diluted with DCM (1000ml), washed with brine (500ml) and the organic phase separated, dried over MgS0₄, filtered and evaporated to give a yellow oil, this was diluted with di-isopropyl ether (300ml) and seeded. The white solid formed was collected, washed with diethyl ether and dried in vacuo to give the title compound white solid, 282g (55%). 1 H NMR (400MHz, CDCI3) δ 1.84-2.00 (1 H, m), 2.32-2.54 (1 H, m), 3.29-3.67 (5H, m) 4.26-4.39 (1 H, m), 4.96-5.26 (3H, m), 6.20-6.84 (1 H, m) 7.29-7.44 (5H, m) LC-MS (method A) Peak at 3.20 mins gave m/e 345 (MH)⁺

Intermediate 62 diethyl 2-{(2R,3S)-1 -[(benzyloxy)carbonyl]-3-r(2,2,2- trifluoroacetyl)amino]pyrrolidin-2-yl}malonate

Diethyl malonate (16.67g, 104mmol, 15.8ml) was dissolved in DCM (90mi) and cooled to -30°C under nitrogen. Tin (iv) chloride (1M in DCM, 52ml, 52mmol) was added maintaining the reaction temperature below -30°C. The reaction mixture was stirred between -30°C and -33°C for 10min. A solution of intermediate 61 (9g, 26mmol) in DCM (90ml) was added maintaining the reaction temperature below -30°C. After the addition was complete the cooling bath was removed and the reaction allowed to warm to 20°C over 1 h. Dilute aqueous hydrochloric acid solution (2N, 90ml) was added and the organic layer separated. The aqueous phase was extracted with DCM (90ml) and the combined organic extracts washed with brine (45ml) dried (Na2S04), and concentrated to give a colourless oil. This was purified by chromatography on silica (Merck 7729) eluting with diethyl etheπcyclohexane (1 :2, then 1:1 , then 2:1 ) to give the title compound as a colourless oil (11.3g)

¹H nmr (400 MHz, CDCI₃): δ 1.23(6H, br t), 1.85-1.96(1 H, m), 2.42-2.53(1 H, m), 3.37-3.45 (1 H, m), 3.72(1 H, br), 4.05-4.24(4H, m), 4.32-4.45(2H, m), 4.6-4.8(1 H, m), 5.14(2H, br s), 7.11 (1 H, br), 7.29-7.4(5H, m)

LC-MS (method A) Peak at 3.45 mins gave m/e 475 (MH)⁺

Intermediate 63 2-Chlorosulphonylpentane

2-Bromopentane (5.73 g, 37.9 mmol) and sodium sulphite (4.89 g, 38.8 mmol) in water (22 mL) were heated under reflux, with vigorous stirring, for 22 hours. The resulting clear solution was allowed to cool to room temperature, then extracted with diethyl ether (2 x 13 mL) and concentrated in vacuo, azeotroping with ethanol and drying in a vacuum oven overnight to give the sodium salt of pentane-2-sulphonic acid (8.69 g). This was treated with phosphorus oxychloride (4 equiv.) and the resulting suspension was heated at 130°C under nitrogen for 5.5 hours. The mixture was allowed to cool to room temperature and treated with ice-water (70 mL), stirred in an ice-water bath for 20 minutes, then extracted with DCM (26 mL). The aqueous phase was re-extracted with DCM (14 mL) and the combined organic extracts were dried over magnesium sulphate and evaporated in vacuo to give the title compound as a yellow liquid (51%). ¹H nmr (CDCI₃): δ 3.68-3.51 , 2.28-2.10 (1 H, m), 1.81-1.33 (7H, m), 1.00 (3H, t)

Intermediate 64: 3S,3aR,6aS)-1 -(1 ,3-benzothiazol-2-yl)-4-[(3- azidopropyl)sulphonyl]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

To a solution of intermediate 59 (46.5 mg, 92 μmol) in dry acetonitrile (3 mL) stirred at room temperature under nitrogen was added sodium azide (108 mg, 1.661 μmol). The mixture was stirred at room temperature for 48 hours. Tic and LC-MS analysis of the reaction mixture showed that starting material was still present. The acetonitrile was removed in vacuo and dry DMF (3 mL) was added. The resulting suspension was stirred at room temperature overnight. The mixture was evaporated in vacuo to a volume of ca. 1 mL. It was then diluted with ethyl acetate (15 mL) and washed with 1 % sodium hydrogen carbonate solution (15 mL). The aqueous phase was re-extracted with ethyl acetate (15 mL) and the combined organic extracts were washed with water (15 mL), dried over magnesium sulphate and evaporated in vacuo to give a yellow oil, which was purified by preparative tic on silica, (eluting with 3:2 cyclohexane:ethyl acetate) to give the title compound as a white solid (32 mg, 82.7%):

LC-MS (method A) Peak at 3.35 mins gave m/e 421 (MH)⁺ Intermediate 65: [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-

2-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-benzothiazol-6-yl]methyl{[2-

(trimethylsilyl)ethyl]sulphonyl}amine

Intermediate 36 (54.3 mg, 77.4 μmol) in dry DCM (750 μL) was treated with TFA

(250 μL) and the solution was stirred at room temperature for four hours. It was then evaporated in vacuo and the residue was partitioned between ethyl acetate (15 mL) and saturaeted aqueous sodium hydrogen carbonate (10 mL). The organic phase was washed with water (10 mL), dried over magnesium sulphate and evaporated in vacuo. The residual gum was purified by preparative tic on silica (3:2 ethyl acetate:cyciohexane eluent) to give the title compound as a white foam (33.7 mg, 72.4%): LC-MS (method A) Peak at 3.67 mins gave m/e 602 (MH)⁺

Synthetic examples:

Example 1 (3S,3aR,6aS)-1 -(1 ,3-benzothiazol-2-yl)-4-(ethyisuiphonyl)-3- methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

Intermediate 21 (3S,3aR,6aS)-1 -Benzothiazol-2-yl-3-methyl-hexahydro-pyrrolo

[3,2-b]pyrrol-2-one trifluoroacetate (17 mg) in dry acetonitrile was treated with triethylamine (14 μL, 2.3 equiv.) followed by ethyl sulphonyl chloride (5 μL, 1.2 equiv.). After sixteen hours at room temperature, the solvent was removed using a stream of nitrogen, and the residue was purified by preparative tic (1 :1 ethyl acetate yclohexane) to give the title compound as a colourless solid (11.7 mg, 73%):

¹H nmr (CDCI₃250 MHz): δ 7.84-7.76 (2H, m), 7.48-7.39 (1 H, m), 7.36-7.26 (1 H, m), 4.30-4.17 (1 H, m), 4.06-3.96 (1 H, m), 3.84-3.65 (2H,m), 3.30-3.04 (4H, m),

2.46-2.17 (1 H, m), 1.46 (3H, t), 1.34 (3H, d),:

LC-MS (method B):

Peak at 4.57 mins gives m/e 366 [MH⁺]

Example 2

Similarly prepared to example 1

(3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-4-(isopropylsulphonyl)-3- methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

colourless solid, 2.1 mg, 12.6% LC-MS (method B):

Peak at 4.69 mins gives m/e 380 [MH⁺]

Example 3

(3S,3aR,6aS)-1 -(1 ,3-benzothiazol-2-yl)-4-[(3-hydroxypropyl) sulphonyl]-3- methylhexahydropyrrolo[3,2-b]pyrrol-2(1H)-one

Intermediate 59 (230 mg) in dry DMF (15 mL) stirred at room temperature under nitrogen was treated with silver nitrate (157 mg) and bis(tributyltin) oxide (520 μL). The mixture was stirred at room temperature for 3 days. The DMF was removed in vacuo and the residue was partitioned between water and ethyl acetate. The organic phase was washed with brine, dried (MgS0₄), and evaporated in vacuo to give a yellow oil. This was partitioned between pentane and acetonitrile to remove tin residues. The acetonitrile phase was evaporated in vacuo to give a gum, which was purified by flash column chromatography on silica (ethyl acetate:cyclohexane 4:1) to give the title compound (slightly contaminated with tin residues) as a colourless solid (63 mg, 35%) after trituration under diethyl ether:

LC-MS (method A):

Peak at time 2.95 minutes gives m/e = 396 (MH)⁺

Example 4

3-[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1 (2H)-yl)sulphonyl]propyl ethylcarbamate

Example 3 (26 mg) in dry THF (3 mL) was treated with triethylamine (9 μL) and ethyl isocyanate (52 μL). The mixture was stirred overight at room temperature then quenched with formic acid (3 drops) and evaporated in vacuo. The mixture was purified by preparative tic followed by dissolution in acetonitrile (2mL) and washing with a solution of potassium fluoride (1 g) in water (1.1 mL) drying (MgS0₄) and evaporation in vacuo to give the title compound as a colourless solid (18 mg, 58%):

¹H nmr (CDCI₃250 MHz): δ 7.80 (2H, m), 7.44 (1 H, m), 7.31 (1 H, m), 4.65 (1 H, broad s), 4.30-4.15 (3H, m), 3.98 (1 H, t), 3.82-3.65 (2H, m), 3.29-3.07 (6H, m), 2.47-2.12 (3H, m), 1.32 (3H, d), 1.15 (3H, t): Mass spec, thermospray: +ve m/e: 467 [MH]⁺

Example 5

Similarly prepared to example 1 (3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1-sulphonic acid dimethylamide colourless solid (66%):

LC-MS (method B):

Peak at 4.69 mins gives m/e 381 [MH⁺]

Example 6

Similarly prepared to example 1

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo 3,2- b]pyrrole-1-sulphonic acid diethylamide colourless solid (61 %):

LC-MS (method B):

Peak at 4.96 mins gives m/e 409 [MH⁺]

(Diethylsulphamoyl chloride was prepared by the method of H Safari and A Blaschette;

Monatsh.Chem.; 101 ; 1970; 1373-1387)

Example 7 Similarly prepared to example 1

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1-sulphonic acid methy-phenyl-amide

colourless solid (14%): LC-MS (method B):

Peak at 3.59 mins gives m/e 443 [MH⁺]

(N-methyl-N-phenylsulphamoyl chloride was prepared by the method of J A Kloek and K L

Lechinsky; J. Org. Chem. 41 4028 (1976)

Example 8

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolof3,2- b]pyrrole-1-sulphonic acid methyl-propyl-amide

Intermediate 21 (20 mg, 0.052 mmol) in anhydrous acetonitrile (0.5 mL) was stirred at room temperature overnight with DIPEA (26 mg, 0.201 mmol, 35 μL) and N-propyl-N-methylsulphamoyl chloride¹ (13 mg, 0.076 mmol). The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane and purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using :2 ethyl acetate:cyclohexane as eluent to give the title compound 17 mg (80%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1H, dd), 7.30 (1H, dd), 4.27-4.12 (1H, m), 3.93-3.80 (1 H, m), 3.79-3.69 (1 H, m), 3.62-3.50 (1 H, m),3.32-3.00 (4H, m), 2.90 (3H, s), 2.43-2.26 (1 H, m), 1.73-1.59 (2H, m), 1.36 (3H, d), 0.95 (3H, t): Mass spec, thermospray: +ve m/e: 409 [MH]⁺ Ref. 1 N-propyi-N-methylsulphamoyl chloride was prepared from N-methyl-N-propylamine using the method of TJ. Cheeseright, A.J. Edwards, D.T. Elmore, J.H. Jones, M. Raissi, E.C. Lewis; J.Chem.Soc.Perkin Trans.1 ; 12; (1994), 1595-1600

Example 8 Alternative synthesis: (3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5- oxo-hexahydro-pyrrolo[3,2-b1pyrrole-1 -sulphonic acid methyl-propyl-amide

Intermediate 26 (200 mg, 0.726 mmol), copper (I) chloride (72 mg, 1 equiv.), freshly-ground anhydrous potassium carbonate (100 mg, 1 equiv.), 2- bromobenzothiazole¹ (233 mg, 1.5 equiv.) and TDA-1 (70.5 mg, 0.3 equiv., 69.8 μL) were stirred in p-xylene (6 mL) under nitrogen and the mixture was heated at reflux for eight hours. The reaction mixture was diluted with ethyl acetate (60 mL) and washed with 0.1 M aqueous hydrochloric acid. The organic phase was separated, dried over anhydrous magnesium sulphate and evaporated under reduced pressure. The crude product was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate:cyclohexane as eluent followed by trituration under cyclohexane to give the title compound (R_f 0.5) 92 mg (31%): ¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.27-4.12 (1 H, m), 3.93-3.80 (1 H, m), 3.79-3.69 (1 H, m), 3.62-3.50 (1 H, m),3.32-3.00 (4H, m),

2.90 (3H, s), 2.43-2.26 (1 H, m), 1.73-1.59 (2H, m), 1.36 (3H, d), 0.95 (3H, t):

Mass spec, thermospray: +ve m/e: 409 [MH]⁺

Ref.1 M.P. Doyle, J.F. Dellaria, B.J.Siegfried, J. Org. Chem. 42, (1977), 2426

Example 9

Similarly prepared to example 8

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolor3,2- b]pyrrole-1 -sulphonic acid isopropylamide colourless solid, 12 mg (30%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1H, dd), 7.30 (1H, dd), 4.30-4.18 (1H, m), 4.11 (1 H, d), 3.92-3.81 (1 H, m), 3.73-3.58 (3H, m), 3.25-3. 03 (2H, m), 2.42-

2.25 (1 H, m), 1.36-1.25 (9H, m):

Mass spec, thermospray: +ve m/e: 395 [MH]⁺ (N-isopropyl-sulphamoyl chloride was prepared from isopropylamine hydrochloride by the method of W L Matier et al., J.Med.Chem.; 15; 1972; 538-541 ;

Example 10 Similarly prepared to example 8

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -sulphonic acid isopropyl-methyl-amide colourless solid, 30 mg (73%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.25-4.12 (2H, m), 3.88-3.79 (1 H, m), 3.78-3.70 (1 H, m), 3.60-3.49 (1 H, m), 3.25-3. 03 (2H, m), 2.71 (3H, s), 2.42-2.25 (1 H, m), 1.32 (3H, d), 1.25 (6H, d): Mass spec, thermospray: +ve m/e: 409 [MH]⁺

Example 11 Similarly prepared to example 8

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -sulphonic acid isobutyl-methyl-amide colourless solid, 12 mg (28%):

Η nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.28-4.16 (1H, m), 4.11 (1 H, d), 3.93-3.82 (1 H, m), 3.80-3.71 (1H, m), 3.62-3.50 (1 H, m) 3.25-3.

00 (4H, m), 2.90 (3H, s), 2.02-1.89(1 H, m), 1.36 (3H, d), 0.95 (6H, d):

Mass spec, thermospray: +ve m/e: 423 [MH]⁺

N-isobutyl-N-methylsulphamoyl chloride was prepared from N-methyl-N-isobutylamine using the method of T.J. Cheeseright, A.J. Edwards, D.T. Elmore, J.H. Jones, M. Raissi, E.C. Lewis; J.Chem.Soc.Perkin Trans.1 ; 12; (1994), 1595-1600

Example 12

(3aS,6S,6aR)-6-Methyl-5-oxo-4-thiazol-2-yl-hexahydro-pyrrolo[3,2-b]pyrrole-1- sulphonic acid isopropyl-methyl-amide

Intermediate 48 (67 mg, 0.243 mmol) copper (I) chloride (24 mg, 1 equiv.), freshly-ground anhydrous potassium carbonate (34 mg, 1 equiv.), 2- bromothiazole (Aldrich) (50 μL, excess) and TDA-1 (23 μL, 0.3 equiv.,) were stirred in p-xylene (5 mL) under nitrogen and the mixture was heated at reflux for ten hours. The solvent was evaporated in vacuo and the residue was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate yclohexane as eluent followed by trituration under cyclohexane to give the title compound (R_f 0.7) 37 mg (42%): ¹H nmr (CDCI₃): δ 7.43 (1 H, d), 7.02 (1 H, d), 4.25-4.04 (2H, m), 3.86-3.76 (1 H, m), 3.73-3.65 (1 H, m), 3.56-3.44 (1 H, m), 3.22-3.09 (1 H, m), 2.97-2.85 (1 H, m), 2.78 (3H, s), 2.37-2.18 (1 H, m), 1.30 (3H, d), 1.22 (6H, d): Mass spec, thermospray: +ve m/e: 359 [MH]⁺

Example 13

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -sulphonic acid allyl-methyl-amide

Intermediate 23 (51 mg, 0.139 mmol) and allyl bromide (24 μL, 2 equiv.) in acetonitrile (1 mL) were stirred with anhydrous caesium carbonate (45 mg, 1 equiv.). The mixture was stirred at room temperature for 24 hours and then filtered. The filtrate was evaporated in vacuo and purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate:cyclohexane as eluent followed by trituration under cyclohexane to give the title compound (R_f 0.3 in ethyl acetate:cyclohexane 1 :4)) 56 mg (63%): Η nmr (CDCI₃): δ 7.84-7.76 (2H, m), 7.47-7.39 (1 H, m), 7.36-7.28 (1 H, m), 5.94- 5.76 (1 H, m), 5.35-5.25 (2H, m), 4.27-4.15 (1 H, m), 3.96-3.71 (4H, m), 3.67-3.53 (1H, m), 3.26-3.03 (2H, m), 2.86 (3H, s), 2.44-2.25 (1H, m), 1.33 (3H, d): Mass spec, thermospray: +ve m/e: 407 [MH]⁺

Example 14 (3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolor3,2- b]pyrrole-1 -sulphonic acid (2,3-dihydroxy-propyl)-methyl-amide

Example 13 (99 mg, 0.244 mmol) was stirred in THF (10 mL) and osmium tetroxide in THF (1.55 mL of a solution containing 40 mg mL^"1, 1.0 equiv.) was added. The mixture was stirred at room temperature for 24 hours then treated with excess aqueous sodium hydrogen sulphite. The mixture was diluted with ethyl acetate, the organic phase was separated, washed with aqueous sodium hydrogen sulphite, dried over anhydrous magnesium sulphate and filtered. The filtrate was evaporated in vacuo and the residue was purified by reverse-phase HPLC to give the title compound 45 mg (41.9%) as a ca. 1 :1 mixture of diastereomers:

Η nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.28 (1 H, m), 4.24-4.16 (1 H, m), 3.98-3.87 (2H, m), 3.78-3.73 (2H, m), 3.67-3.56 (2H, m), 3.46-3.30 (2H, m), 3.23-3.15 (1 H, s), 3.12-3.04 (1 H, m), 3.03 (3H, s), 2.76 (1H, broad s), 2.42-2.30 (1 H, m), 2.29 (1 H, broad s), 1.35 (3H, d): Mass spec, thermospray: +ve m/e: 441 [MH]⁺

Example 15 and example 16 separated isomers of (3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo- hexahydro-pyrrolo[3,2-b]pyrrole-1 -sulphonic acid (2,3-dihydroxy-propyl)-methyl- amide

The 1 :1 mixture of diastereomers of example 14 (3aS,6S,6aR)-4-Benzothiazol- 2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2-b1pyrrole-1 -sulphonic acid (2,3- dihydroxy-propyl)-methyl-amide (40 mg) was separated by chromatography on a chiral column to give: first peak

Example 15 (17 mg):

¹H nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.28 (1H, m), 4.24-4.16 (1H, m), 3.98-3.87 (2H, m), 3.78-3.73 (2H, m), 3.67-3.56 (2H, m), 3.46-3.30 (2H, m), 3.23-3.15 (1 H, s), 3.12-3.04 (1 H, m), 3.03 (3H, s), 2.76 (1 H, broad s), 2.42-2.30 (1 H, m), 2.29 (1 H, broad s), 1.35 (3H, d): Mass spec, thermospray: +ve m/e: 441 [MH]⁺

second peak Example 16: ¹H nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.28

(1H, m), 4.24-4.16 (1 H, m), 3.98-3.87 (2H, m), 3.78-3.73 (2H, m), 3.67-3.56 (2H, m), 3.39-3.35 (2H, m), 3.23-3.15 (1 H, s), 3.12-3.04 (1 H, m), 3.03 (3H, s), 2.76 (1 H, broad s), 2.42-2.30 (1 H, m), 2.29 (1 H, broad s), 1.35 (3H, d): Mass spec, thermospray: +ve m/e: 441 [MH]⁺ Example 17

2- (4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- blpyrrole-1 -sulphonyl )-methyl-amino]-N,N-dimethyl-acetamide

A 2mL aliquot of a solution containing intermediate 23 (420 mg, 1.146 mmol) in

1 :1 acetonitrile:dichloromethane (20 mL) was stirred with caesium carbonate (53 mg) and N,N-dimethyl chloroacetamide (18 mg, 1.3 equiv., 16.3 μL) overnight at room temperature. The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane and purified by preparative plate chromatography

(Whatman PK6F silica gel 60A plate) using 1 :1 ethyl acetate:cyclohexane as eluent to give the title compound (R_f 0.2) 50 mg (96.6%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.25-4.07 (3H, m), 3.91-3.70 (3H, m), 3.23-3.10 (1 H, m), 3.10-2.95 (10H, m), 2.48-2.20 (1 H, m),

1.35 (3H, d):

Mass spec, thermospray: +ve m/e: 452 [MH]⁺

Example 18

Similarly prepared to example 17

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-acetamide

(using bromoacetamide (21 mg) as the electrophile) colourless solid, 41 mg (84.4%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 6.3 (1 H, broad s), 5.68 (1H, broad s), 4.26-4.14 (1H, m), 4.01-3.84 (3H, m), 3.84-3.77 (1H, m),

3.71-3.58 (1 H, m), 3.24-3.01 (5H, m), 2.47-2.19 (1 H, m), 1.35 (3H, d):

Mass spec, thermospray: +ve m/e: 424 [MH]⁺

Example 19

Similarly prepared to example 17

4-f(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- blpyrrole-1-sulphonyl)-methyl-amino1-but-2E-enoic acid methyl ester (using methyl (E)-bromocrotonate (32 mg, 21 μL ) as the electrophile) colourless solid, 44 mg (82.6%):

Η nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 6.96-6.84 (1 H, m), 6.11-6.02 (1 H, m), 4.26-3.96 (3H, m), 3.95-3.84 (1 H, m), 3.80-3.72 (4H, m),

3.66-3.54 (1 H, m), 3.25-3.14 (2H, m), 2.70 (3H, s), 2.45-2.17 (1 H, m), 1.35 (3H, d):

Mass spec, thermospray: +ve m/e: 465 [MH]⁺

Example 20

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -sulphonic acid (2-diethylamino-ethyl)-methyl-amide formate

A 2mL aliquot of a solution containing intermediate 23 (420 mg, 1.146 mmol) in 1 :1 acetonitrile.-dichloromethane (20 mL) was stirred with caesium carbonate (53 mg) and 2-bromoethyl diethylamine hydrobromide (39 mg) overnight at room temperature.

The mixture was diluted with ethyl acetate (50 mL) and washed with saturated aqueous sodium hydrogen carbonate (10 mL). The organic phase was separated, dried over anhydrous magnesium sulphate and evaporated under reduced pressure. The crude product was purified by reverse-phase HPLC (protocol fcal2) to give the title compound as the formate salt 20 mg (34.1%): H nmr (CDCI₃): δ 8.40 (1 H, broad s) 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.26-4.15 (1 H, m), 3.91-3.55 (5H, m), 3.25-3.04 (8H, m), 2.98 (3H, s), 2.45- 2.26 (1 H, m), 1.38-1.24 (9H, d):

Mass spec, thermospray: +ve m/e: 466 [MH]⁺

Example 21 Similarly prepared to example 17

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- blpyrrole-1 -sulphonic acid methyl-pyridin-2-ylmethyl-amide (using 2-chloromethyl pyridine hydrochloride (25 mg) as the electrophile) colourless solid, 17 mg (32.4%): Η nmr (CDCI₃): δ 8.59 (1 H, d), 7.85-7.70 (3H, m), 7.49-7.40 (2H, m), 7.36-7.22 (2H, m), 4.67-4.46 (2H, ABq), 4.27-4.16 (1 H, m), 3.99-3.88 (1 H, m), 3.85-3.77 (1 H, m), 3.69-3.57 (1 H, m), 3.26-3.14 (2H, m), 2.94 (3H, s), 2.45-2.17 (1 H, m), 1.35 (3H, d): Mass spec, thermospray: +ve m/e: 458 [MH]⁺

Example 22

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolof3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-propionic acid methyl ester

Intermediate 23 (20 mg, 0.055 mmol) and methyl 2-bromopropionate (6.7 μL, 1.1 equiv.) in 1 :1 acetonitrile: dichloromethane (0.5 mL) were stirred with anhydrous caesium carbonate (20 mg, 1.1 equiv.). The mixture was stirred at room temperature overnight. The mixture was filtered, the filtrate was evaporated in vacuo and the residue was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate:cyclohexane as eluent to give the title compound (R_f 0.55 in ethyl acetate:cyclohexane 1 :1) 22 mg (89%) as a ca. 1 :1 mixture of two diastereomers:

¹H nmr (CDCI₃): δ 7.84-7.76 (2H, m), 7.47-7.39 (1 H, m), 7.35-7.27 (1 H, m), 4.77- 4.59 (1 H, m), 4.26-4.09 (1 H, m), 3.89-3.59 (6H, m), 3.26-2.99 (2H, m), 2.98 and 2.93 (3H, 2s), 2.43-2.25 (1 H, m), 1.50 (3H, d), 1.34 and 1.29 (3H, 2d): Mass spec, thermospray: +ve m/e: 453 [MH]⁺

Example 23

(3aS,6S,6aR)-4-Benzothiazol-2-yl-6-methyl-5-oxo-hexahydro-pyrrolo[3,2- b]pyrrole-1 -sulphonic acid sec-butyl-methyl-amide

Intermediate 23 (20 mg, 0.055 mmol) and 2-iodobutane (1.1 mL, large excess) in acetonitrile (1 mL) were stirred with anhydrous caesium carbonate (18 mg, 1 equiv.). The mixture was stirred at room temperature for 72 hours. The solvent was evaporated in vacuo and the residue was taken up in ethyl acetate and washed with saturated aqueous sodium hydrogen carbonate. The organic phase was separated, dried over anhydrous magnesium sulphate, and evaporated in vacuo to give a yellow gum, which was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate:cyclohexane as eluent followed by trituration under cyclohexane to give the title compound (R_f 0.45 in ethyl acetate:cyclohexane 1 :2)) 17 mg (74%) as a ca. 1:1 mixture of two diastereomers:

¹H nmr (CDCI₃): δ 7.84-7.76 (2H, m), 7.47-7.39 (1 H, m), 7.36-7.28 (1 H, m), 4.27- 4.15 (1H, m), 3.96-3.69 (3H, m), 3.61-3.47 (1H, m), 3.26-3.13 (1H, m), 3.11-2.99 (1 H, m), 2.75 and 2.73 (3H, 2s), 2.44-2.25 (1 H, m), 1.61-1.43 (2H, m), 1.33 (3H, d), 1.21 and 1.20 (3H, 2d), 0.95 and 0.94 (3H, 2t): Mass spec, thermospray: +ve m/e: 423 [MH]⁺

Example 24 Similarly prepared to example 17

[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-acetic acid tert-butyl ester

(using t-butyl bromoacetate (30 mg, 21 μL) as the electrophile) colourless solid, 55 mg (100%): ¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.43 (1 H, dd), 7.30 (1 H, dd), 4.23-4.11 (1 H, m), 4.04-3.67 (5H, m), 3.24-2.99 (5H, m), 2.45-2.17 (1 H, m), 1.49 (9H, s), 1.35

(3H, d):

Mass spec, thermospray: +ve m/e: 481 [MH]⁺

Example 25

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- b]pyrrole-1 -sulphonyl )-methyl-amino1-N-benzyl-acetamide

A 1mL aliquot of a solution containing the active ester {from intermediate 24

(160 mg, 0.377 mmol), HATU (150 mg, 1.05 equiv.) HOAT (54 mg, 1.05 equiv.) and DIPEA (64 μL) in dry DMF (8 mL)} was stirred with benzylamine (5.2 μL) at room temperature overnight. The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane (5 mL) and stirred with saturated aqueous sodium hydrogen carbonate (2 mL). The mixture was filtered through a hydrophobic frit and the organic filtrate was evaporated using a stream of nitrogen. The residual gum was taken up in dichloromethane and purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using ethyl acetate as eluent to give the title compound (R_f 0.6) as a colourless solid 17 mg (70.2%):

¹H nmr (CDCI₃): δ 7.85-7.75 (2H, m), 7.46-7.25 (7H, m), 6.65 (1 H, broad t), 4.49 (2H, d), 4.18-4.06 (1 H, m), 4.04-3.83 (3H, m), 3.80-3.72 (1 H, m), 3.66-3.54 (1 H, m), 3.23-2.98 (5H, m), 2.45-2.16 (1 H, m), 1.29 (3H, d): Mass spec, thermospray: +ve m/e: 514 [MH]⁺

Example 26

Similarly prepared to example 25

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-N-thiazol-2-yl-acetamide colourless solid 12 mg (50.3%):

¹H nmr (CDCI₃): δ 7.85-7.76 (2H, m), 7.52 (1 H, d), 7.48-7.39 (1 H, m), 7.36-7.28

(1 H, m), 7.07 (1 H, d), 4.38-4.16 (3H, m), 3.98-3.70 (3H, m), 3.25-3.05 (5H, m),

2.48-2.31 (1 H, m), 1.31 (3H, d): Mass spec, thermospray: +ve m/e: 507 [MH]⁺

Example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1 -sulphonyl )-methyl-amino]-N-thiophen-2-ylmethyl-acetamide

A 1mL aliquot of a solution containing the active ester {from intermediate 24 (340 mg, 0.377 mmol), PyAOP (439 mg, 1.05 equiv.) HOAT (115 mg, 1.05 equiv.) and DIPEA (64 μL) in dry DMF (20 mL)} was stirred with 2- thienylmethylamine (4.5 μL) at room temperature for 48 hours.

The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane (5 mL) and stirred with saturated aqueous sodium hydrogen carbonate (2 mL). The mixture was filtered through a hydrophobic frit and the organic filtrate was evaporated using a stream of nitrogen. The residual gum was taken up in dichloromethane and purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using ethyl acetate:cyclohexane (1 :1 ) as eluent to give the title compound (R_f 0.5) as a colourless solid 7 mg (33.7%):

¹H nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.29

(1H, m), 7.25-7.23 (1H, dd), 7.01-6.98 (1 H, m), 6.97-6.95 (1H, m), 6.68 (1H, broad t), 4.67 (2H, d), 4.18-4.12 (1 H, m), 4.00-3.86 (3H, m), 3.80-3.75 (1 H, m),

3.64-3.57 (1 H, m), 3.21-3.13 (1 H, m), 3.11-3.04 (1 H, m), 3.01 (3H,s), 2.43-2.31

(1 H, m), 1.30 (3H, d):

Mass spec, thermospray: +ve m/e: 520 [MH]⁺

Example 28

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- blpyrrole-1-sulphonyl)-methyl-amino1-N-(4-methoxy-benzyl)-acetamide colourless solid 9.0 mg (41.4%):

¹H nmr (CDCI₃) (400 MHz): δ 7.83-7.77 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.29

(1H, m), 7.23-7.19 (2H, m), 6.89-6.85 (2H, m), 6.59 (1H, broad t), 4.42 (2H, d),

4.16-4.08 (1 H, m), 4.00-3.84 (3H, m), 3.78-3.74 (4H, m), 3.62-3.55 (1 H, m), 3.20-3.12 (1 H, m), 3.08-3.01 (1 H, m), 3.00 (3H,s), 2.41-2.29 (1 H, m), 1.29 (3H, d):

Mass spec, thermospray: +ve m/e: 544 [MH]⁺

Example 29

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1 -sulphonyl )-methyl-amino]-N-(4-chloro-benzyl)-acetamide colourless solid 10 mg (45.6%): ¹H nmr (CDCI₃) (400 MHz): δ 7.83-7.77 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.29 (3H, m), 7.25-7.21 (2H, m), 6.73 (1 H, broad t), 4.47 (2H, d), 4.21-4.13 (1H, m), 4.01-3.85 (3H, m), 3.78-3.74 (1 H, m), 3.63-3.56 (1 H, m), 3.21-3.13 (1 H, m), 3.11-3.04 (1 H, m), 3.00 (3H,s), 2.42-2.31 (1 H, m), 1.30 (3H, d): Mass spec, thermospray: +ve m/e: 548/550 [MH]⁺ Example 30

Similarly prepared to example 27

2-r(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolof3,2- b1pyrrole-1-sulphonyl)-methyl-amino]-N-(4-trifluoromethyl-benzyl)-acetamide colourless solid 9.0 mg (38.7%):

¹H nmr (CDCI₃) (400 MHz): δ 7.83-7.77 (2H, m), 7.63-7.59 (2H, m), 7.46-7.39 (3H, m), 7.34-7.29 (1 H, m), 6.83 (1 H, broad t), 4.56 (2H, d), 4.23-4.15 (1H, m), 4.03-3.86 (3H, m), 3.81-3.77 (1 H, m), 3.64-3.57 (1H, m), 3.22-3.14 (1H, m), 3.13-3.05 (1 H, m), 3.02 (3H,s), 2.44-2.33 (1 H, m), 1.31 (3H, d): Mass spec, thermospray: +ve m/e: 582 [MH]⁺

Example 31

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1 -sulphonyl )-methyl-amino]-N-(4-sulphamoyl-benzyl)-acetamide colourless solid 8.0 mg (33.7%):

¹H nmr (CDCI₃) (400 MHz): δ 7.89-7.86 (2H, m), 7.84-7.78 (2H, m), 7.47-7.40

(3H, m), 7.35-7.30 (1H, m), 6.98 (1 H, broad t), 5.04 (2H, broad s), 4.61-4.49 (2H, m), 4.21-4.13 (1 H, m), 4.04-3.92 (2H, ABq), 3.90-3.84 (1 H, m), 3.79-3.75 (1 H, m), 3.63-3.56 (1 H, m), 3.18-3.02 (5H, m), 2.42-2.30 (1 H, m), 1.29 (3H, d):

Mass spec, thermospray: +ve m/e: 593 [MH]⁺

Example 32

Similarly prepared to example 27 2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-N-(2-hydroxy-1R-phenyl-ethyl)-acetamide colourless solid 10 mg (46%):

¹H nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.47-7.28 (7H, m), 7.05 (1H, broad d), 5.14-5.09 (1 H, m), 4.20-4.12 (1 H, m), 4.02-3.85 (4H, m), 3.81-3.76 (1 H, m), 3.66-3.59 (1 H, m), 3.22-3.15 (1 H, m), 3.08-3.01 (4H, m), 2.40-2.28 (2H, m), 1.30 (3H, d):

Mass spec, thermospray: +ve m/e: 544 [MH]⁺

Example 33 Similarly prepared to example 27 2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolof3,2- blpyrrole-1-sulphonyl)-methyl-amino1-N-(2-hydroxy-1 S-phenyl-ethyl)-acetamide colourless solid 10 mg (46%):

Η nmr (CDCI₃) (400 MHz): δ 7.83-7.77 (2H, m), 7.46-7.26 (7H, m), 7.03 (1 H, broad d), 5.13-5.08 (1 H, m), 4.21-4.13 (1 H, m), 4.02-3.85 (4H, m), 3.80-3.75

(1 H, m), 3.66-3.58 (1 H, m), 3.22-3.15 (1H, m), 3.10-3.03 (4H, m), 2.41-2.29 (2H, m), 1.32 (3H, d):

Mass spec, thermospray: +ve m/e: 544 [MH]⁺

Example 34

Similarly prepared to example 27

2-r(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- blpyrrole-1-sulphonyl)-methyl-amino]-N-(5-bromo-furan-2-ylmethyl)-acetamide colourless solid 4.0 mg (17.2%): Η nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.29

(1H, m), 6.63 (1H, broad t), 6.27-6.24 (2H, m), 4.51-4.40 (2H, d), 4.20-4.13 (1H, m), 4.00-3.87 (3H, m), 3.82-3.77 (1 H, m), 3.66-3.59 (1 H, m), 3.22-3.14 (1 H, m), 3.13-3.05 (1 H, m), 3.02 (3H, s), 2.44-2.32 (1 H, m), 1.31 (3H, d): Mass spec, thermospray: +ve m/e: 582/584 [MH]⁺

Example 35

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-N-(4-methyl-thiazol-2-ylmethyl)-acetamide colourless solid 6 mg (20.9%):

Η nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.46-7.41 (1 H, m), 7.34-7.29 (1H, m), 7.09 (1H, broad t), 6.83 (1H, m), 4.77 (2H, d), 4.21-4.13 (1H, m), 4.04- 3.88 (3H, m), 3.82-3.77 (1 H, m), 3.71-3.63 (1 H, m), 3.22-3.14 (1 H, m), 3.12-3.03 (4H, m), 2.44-2.32 (4H, m), 1.32 (3H, d): Mass spec, thermospray: +ve m/e:.535 [MH]⁺

Example 36

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2- blpyrrole-1-sulphonyl)-methyl-amino1-N-pyridin-3-ylmethyl-acetamide colourless gum 9.0 mg (43.7%):

Η nmr (CDCI₃) (400 MHz): δ 8.58-8.54 (2H, m), 7.84-7.78 (2H, m), 7.66-7.63 (1H, m), 7.46-7.41 (1 H, m), 7.34-7.27 (2H, m), 6.86 (1 H, broad t), 4.52 (2H, d), 4.21-4.13 (1 H, m), 4.02-3.86 (3H, m), 3.80-3.75 (1 H, m), 3.66-3.58 (1 H, m), 3.21-3.07 (2H, m), 3.01 (3H, s), 2.43-2.31 (1 H, m), 1.30 (3H, d):

Mass spec, thermospray: +ve m/e: 515 [MH]⁺

Example 37

Similarly prepared to example 27 2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-N-pyridin-2-ylmethyl-acetamide colourless gum 4.6 mg (22.3%): nmr (CDCI₃) (400 MHz): δ 8.56-8.54 (1 H, m), 7.84-7.78 (2H, m), 7.71-7.67

(1H, m), 7.54 (1 H, broad t), 7.46-7.41 (1 H, m), 7.34-7.29 (1 H, m), 7.28-7.26 (1H, m), 7.24-7.20 (1 H, m), 4.61 (2H, d), 4.24-4.17 (1 H, m), 4.04-3.93 (3H, m), 3.85-

3.80 (1 H, m), 3.78-3.70 (1 H, m), 3.25-3.17 (1 H, m), 3.11-3.05 (4H, m), 2.45-2.33

(1H, m), 1.33 (3H, d):

Mass spec, thermospray: +ve m/e: 515 [MH]⁺

Example 38

Similarly prepared to example 27

2-[(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- b]pyrrole-1 -sulphonyl)-methyl-amino]-N-(1 -methyl-1 H-tetrazol-5-ylmethyl)- acetamide colourless gum 6.5 mg (31.3%):

Η nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.57 (1 H, broad t), 7.46-7.41 (1 H, m), 7.34-7.29 (1 H, m), 4.76 (2H, d), 4.24-4.15 (4H, m), 4.04-3.86 (3H, m), 3.81-3.76 (1 H, m), 3.68-3.61 (1 H, m), 3.20-3.07 (2H, m), 3.02 (3H, s), 2.43-2.32 (1H, m), 1.30 (3H, d): Mass spec, thermospray: +ve m/e: 520 [MH]⁺

Example 39

Similarly prepared to example 27 2-r(4-Benzothiazol-2-yl-6S-methyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolor3,2- b]pyrrole-1-sulphonyl)-methyl-amino]-N-(3-methoxy-isothiazol-5-ylmethyl)- acetamide colourless solid 4.6 mg (28.0%): nmr (CDCI₃) (400 MHz): δ 7.84-7.78 (2H, m), 7.46-7.41 (1H, m), 7.34-7.29

(1H, m), 6.95 (1H, broad t), 6.50 (1H, s), 4.65 (2H, d), 4.23-4.15 (1H, m), 4.03-

3.87 (6H, m), 3.81-3.76 (1 H, m), 3.66-3.58 (1 H, m), 3.21-3.13 (1 H, m), 3.13-3.05

(1H, m), 3.02 (3H, s), 2.44-2.32 (1H, m), 1.32 (3H, d):

Mass spec, thermospray: +ve m/e: 551 [MH]⁺

Example 40

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulfonyl](methyl)amino]-N-[(1 S)-1 - benzyi-2-hydroxyethyl]acetamide

Intermediate 24 (20 mg, 47.11 μmol), EDC (10 mg,. 1.1 equiv.) and HOBT (7 mg, 1.1 equiv.) were atirred in anhydrous acetonitrile (1 mL) at 0°C and DIPEA (13.4 mg, 2.2 equiv., 18μL) was added. The mixture was stirred at 0°C for 30 minutes, then treated with (S)-2-amino-3-phenylpropanol (8 mg, 1.1 equiv.).

The homogeneous mixture was left at room temperature overnight, the solvent was removed by a stream of nitrogen, and the residue was taken up in DCM (0.5 mL) and washed with 0.1 M HCI aq. then with saturated aqueous sodium hydrogen carbonate. The organic phase was separated after each washing using a hydrophobic frit. The crude product was purified by preparative tic to give the title compound as a colourless solid (20 mg).: Η nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1 H, m), 7.35-7.28 (3H, m), 7.27-7.20 (3H, m), 6.53 (d, 1 H), 4.29-4.10 (2H, m), 3.90-3.70 (5H, m), 3.68-3.60 (1 H, m), 3.60-3.52 (1 H, m), 3.21-3.12 (1 H, m), 3.12-3.04 (1 H, m), 2.99-2.82 (2H, m), 2.79 (3H, s), 2.54 (1 H, broad s), 2.42-2.30 (1 H, m), 1.29 (3H, d):

Mass spec, thermospray: +ve m/e: 558 [MH]⁺

Example 41 Similarly prepared to example 40 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-[(1 R)-1 - benzyl-2-hydroxyethyl]acetamide

Η nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1H, m), 7.35-7.28 (3H, m), 7.27-7.20 (3H, m), 6.54 (d, 1 H), 4.29-4.11 (2H, m), 3.97-3.70 (5H, m), 3.68-3.54 (2H, m), 3.21-3.13 (1 H, m), 3.10-3.03 (1 H, m), 2.99-2.82 (2H, m), 2.78 (3H, s), 2.61 (1H, broad s), 2.41-2.29 (1H, m), 1.29 (3H, d): Mass spec, thermospray: +ve m/e: 558 [MH]⁺

Example 42 Similarly prepared to example 40

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-[(1R,2S)-

2-hydroxy-1 ,2-diphenylethyl]acetamide

colourless solid (25 mg, 85%): nmr (CDCI₃) (400 MHz): δ 7.84-7.76 (2H, m), 7.46-7.41 (1 H, m), 7.35-7.28 (3H, m), 7.28-7.22 (6H, m), 7.18 (1 H, d), 7.13-7.06 (4H, m), 5.28-5.23 (1 H, m), 5.08-5.04 (1H, m), 4.18-4.09 (1H, m), 3.88-3.80 (3H, m), 3.79-3.73 (1H, m), 3.60-3.52 (1 H, m), 3.21-3.12 (1 H, m), 3.07-2.99 (1 H, m), 2.89 (3H, s), 2.6-6 (1H, broad s), 2.39-2.27 (1 H, m), 1.28 (3H, d):

Mass spec, thermospray: +ve m/e: 620 [MH]⁺

Example 43 Similarly prepared to example 40 methyl (2R)-2-({2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yi)sulphonyl](methyl) amino]acetyl}amino)-3-hydroxypropanoate

colourless solid (20 mg, 80%): Η nmr (CDCI₃) (400 MHz): δ 7.83-7.75 (2H, m), 7.46-7.41 (1 H, m), 7.35-7.28 (1 H, m), 7.19 (1 H, d), 4.72-4.67 (1 H, m), 4.27-4.18 (1 H, m), 4.12-4.04 (2H, m), 3.98-3.88 (3H, m), 3.82-3.77 (4H, m), 3.76-3.68 (1 H, m), 3.23-3.15 (1 H, m), 3.133.02 (4H, m), 2.59 (1 H, broad t), 2.43-2.33 (1H, m), 1.32 (3H, d): Mass spec, thermospray: +ve m/e: 526 [MH]⁺

Example 44

Similarly prepared to example 40 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-[(1 R)-1 • (hydroxymethyl)-2-methylpropyl]acetamide

colourless solid (17 mg, 76%): LC-MS (method A):

Peak at 3.15 mins gives m/e 510 [MH⁺]

Example 45 Similarly prepared to example 40

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazoI-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-[(1S)-2- cyclohexyl-1-(hydroxymethyl)ethyl]acetamide

colourless solid (19 mg, 70%): LC-MS (method A): Peak at 3.40 mins gives m/e 564 [MH⁺]

Example 46

Similarly prepared to example 40

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-[(1S)-2- hydroxy-1-(4-hydroxybenzyl)ethyl]acetamide colourless solid (16 mg, 59%):

LC-MS (method A):

Peak at 3.12 mins gives m/e 574 [MH⁺]

Example 47

2-[[((3aS ,6S ,6aR)-4-( 1 ,3-benzothiazol-2-yl )-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(1H- pyrazol-3-yl)acetamide

Intermediate 24 (20 mg, 47.11 μmol) was stirred in dry DCM (1 mL). Triethylamine (6.55 μL) was added and the mixture was stirred until homogeneous, then treated with oxalyl chloride (4.11 μL) (effervescence). The mixture was stirred at room temperature for 15 minutes, then treated with 3- aminopyrazole (3.7 mg, 1.1 equiv.). The homogeneous mixture was left at room temperature overnight, the solvent was removed by a stream of nitrogen, and the residue was purified by preparative tic (ethyl acetate) to give the title compound as a colourless solid (11 mg).: LC-MS (method A): Peak at 3.19 mins gives m/e 490 [MH⁺]

Example 48

Similarly prepared to example 47 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(1,3,4- thiadiazol-2-yl)acetamide

colourless solid (5 mg, 21%): LC-MS (method A):

Peak at 3.16 mins gives m/e 508 [MH⁺]

Example 49 (R3746/71/1 ) 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-cyclobutyl acetamide

Intermediate 24 (20 mg, 47 μmol) was stirred in dry acetonitrile (1 mL) at room temperature. Then EDC (10 mg, 52 μmol, 1.1 equiv) was added to the stirred mixture followed by HOBT (9.5 mg, 70 μmol, 1.5 equiv) and then DIPEA (18 μL, 103 μmol, 2.2 equiv) whereupon the mixture became a homogeneous solution. Cyclobutylamine (4.42 μL, 52 μmol, 1.1 equiv) was added to the mixture, which was stirred for 24 hours before a solution of EDC (10 mg), DIPEA (9 μL, 52 μmol, 1.1 equiv) and cyclobutylamine (4.42 μL, 52 μmol, 1.1 equiv) in dry acetonitrile (0.5 mL) was added. The mixture was stirred a further 19 hours 30 minutes before it was blown to dryness under a stream of nitrogen. The residue was dissolved in dichloromethane (1 mL) and washed with 0.1 M hydrochloric acid (0.5 mL) followed by saturated aqueous sodium bicarbonate solution (0.5 mL). The organic phase was isolated using a hydrophobic frit and evaporated. The crude product was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :1 cyclohexane: ethyl acetate as eluent to give the title compound as a white solid, 10.3 mg (45%): Η nmr (CDCI₃): δ 7.80 (2H, m), 7.44 (1 H, m), 7.31 (1 H, m), 6.40 (1 H, d), 4.41 (1 H, m), 4.19 (1 H, td), 3.86 (4H, m), 3.64 (1H, m), 3.19 (1 H, m), 3.10 (1 H, m), 3.00 (3H, s), 2.37(3H, m), 1.92 (2H, m), 1.75 (2H, m), 1.33 (3H, d): LC/Mass spec, electrospray: +ve m/e: 478 [MH]⁺:

Example 50

Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonylJ(methyl)amino]-N-(1-methylbutyl)acetamide

nmr (CDCI₃): δ 7.80 (2H, m), 7.43 (1 H, m), 7.31 (1 H, m), 6.07 (1 H, m), 4.20 (1 H, td), 4.02 (1 H, m), 3.89 (3H, m), 3.80 (1 H, dd), 3.64 (1H, m), 3.19 (1H, m), 3.10 (1H, m), 3.00 (3H, d), 2.39 (1 H, m), 1.45 (2H, m), 1.35 (2H, m), 1.33 (3H, d), 1.17 (3H, d), 0.92 (3H, t): LC/Mass spec, electrospray: +ve m/e: 494 [MH]⁺: Example 51

Similarly prepared to example 49 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazoi-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-cyclopentylacetamide

nmr (CDCI₃): δ 7.80 (2H, m), 7.44 (1 H, m), 7.31 (1 H, m), 6.20 (1 H, d), 4.21 (2H, m), 3.87 (3H, m), 3.80 (1 H, dd), 3.64 (1 H, m), 3.19 (1 H, m), 3.10 (1 H, m), 3.00 (3H, s), 2.38(1 H, m), 2.01 (2H, m), 1.65 (4H, m), 1.43 (2H, m), 1.33 (3H, d): LC/Mass spec, electrospray: +ve m/e: 492 [MH]⁺:

Example 52 Similarly prepared to example 49

(3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-N,6-dimethyl-N-[2-(2-methylpiperidin-1-yl)- 2-oxoethyl]-5-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Η nmr (CDCI₃): δ 7.80 (2H, m), 7.43 (1 H, m), 7.29 (1H, m), 4.84 (0.5H, bs), 4.42 (0.5H, bs), 4.14 (3H, m), 3.85 (2H, m), 3.76 (1 H, m), 3.48 (2H, q), 3.16 (1H, m), 3.06 (3H, s), 3.03 (1 H, m), 2.38 (1 H, m), 1.63 (4H, m), 1.34 (3H, d), 1.20(5H, m): LC/Mass spec, electrospray: +ve m/e: 506 [MH]⁺:

Example 53

Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(tert-butyl)acetamide nmr (CDCI₃): δ 7.80 (2H, m), 7.43 (1 H, m), 7.31 (1 H, m), 6.02 (1 H, s), 4.18 (1 H, m), 3.90 (1 H,m), 3.79 (3H, m), 3.65 (1 H, m), 3.18 (1 H, m), 3.09 (1 H, m), 3.00 (3H, s), 2.38 (1 H, m), 1.38 (9H, s), 1.33 (3H, d): LC/Mass spec, electrospray: +ve m/e: 480 [MH]⁺:

Example 54 Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(1 ,1-dimethylprop-2-ynyl)acetamide Η nmr (CDCI₃): δ 7.80 (2H, m), 7.44 (1 H, m), 7.31 (1 H, m), 6.32 (1 H, s), 4.20 (1 H, m), 3.91 (1 H,m), 3.85 (2H, d), 3.80 (1 H, dd), 3.66 (1 H, m), 3.19 (1H, m), 3.10 (1H, m), 3.03 (3H, s), 2.39 (1 H, m), 2.37 (1 H, s), 1.67 (6H, s), 1.33 (3H, d): LC/Mass spec, electrospray: +ve m/e: 490 [MH]⁺:

Example 55

Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(1 ,2-dimethylpropyl)acetamide

Η nmr (CDCI₃): δ 7.80 (2H, m), 7.43 (1 H, m), 7.31 (1 H, m), 6.15 (1 H, bd), 4.19 (1H, m), 3.89 (4H,m), 3.79 (1 H, m), 3.64 (1H, m), 3.18 (1H, m), 3.10 (1H, m), 3.01 (3H, s), 2.38 (1 H, m), 1.74 (1 H, m), 1.33 (3H, d), 1.12 (3H, d), 0.92 (6H, d): LC/Mass spec, electrospray: +ve m/e: 494 [MH]⁺:

Example 56

Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(2-methoxy-1-methylethyl)acetamide Η nmr (CDCI₃): δ 7.80 (2H, m), 7.43 (1 H, m), 7.31 (1 H, m), 6.40 (1 H, bt), 4.19 (2H, m), 3.89 (3H, m), 3.80 (1H, dd), 3.66 (1 H, m), 3.39 (2H, m), 3.37 (3H, s), 3.19 (1H, m), 3.10 (1 H, m), 3.00 (3H, s), 2.39 (1 H, m), 1.33 (3H, d), 1.22 (3H, d): LC/Mass spec, electrospray: +ve m/e: 496 [MH]⁺:

Example 57

Similarly prepared to example 49

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(sec-butyl)acetamide Η nmr (CDCI₃): δ 7.80 (2H, m), 7.44 (1 H, m), 7.31 (1 H, m), 6.06 (1 H, m), 4.19 (1 H, m), 3.91 (4H, m), 3.80 (1 H, dd), 3.64 (1 H, m), 3.19 (1 H, m), 3.11 (1 H, m), 3.01 (3H, s), 2.39 (1 H, m), 1.51 (2H, m), 1.33 (3H, d), 1.17 (3H, d), 0.93 (3H, t): LC/Mass spec, electrospray: +ve m/e: 480 [MH]⁺:

Example 58

2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5-oxohexahydropyrrolo[3,2- b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-ethyl-N-isopropylacetamide

Intermediate 24 (20.3mg, O.Oδmmol), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride, (9.8mg, O.Oδmmol), di-isopropylethylamine (17.9μl, 0.103mmol) and 1-hydroxybenzotriazole (9.5mg, 0.07mmol) were dissolved in acetonitrile. N-ethyl-N-isopropyl amine (16.9μl, 0.14mmol) was added and the mixture stirred at room temperature for 40 hours. A further addition of 1-(3- dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (26.8mg, 0.14mmol) and di-isopropylethylamine (24.4μl, 0.14mmol) in acetonitrile (100μl) was made. After 24 hours the solvent was removed with a stream of nitrogen, the residue taken up in dichloromethane (1 ml) and washed with 0.1 M aqueous hydrochloric acid (0.5ml), separated then washed with saturated aqueous sodium hydrogen carbonate (1mi) passed through a hydrophobic frit and the dichloromethane removed from the collected organic phase with a stream of nitrogen. The residual gum was taken up in dichloromethane and purified using preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :1 ethyl aecetate:cyclohexane as eluant to give the title compound as a clear gum, 15.9mg (67%): Η NMR (400MHZ, CDCI₃) δ 1.12-1.17 (11 H, m), 1.35 (3H, d), 2.39 (1 H, m), 2.98-3.34 (8H, m), 3.74-3.96 (4H, m), 4.05-4.26 (4H, m), 7.30 (1 H, td), 7.43 (1H, td), 7.80 (2H, m) LC-MS (A, 5.5min)

Peak at time 3.47 minutes gave m/e 494 (M+H)⁺

Example 59 Similarly prepared to example 58 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methy!-5-oxohexahydropyrrolo[3,2- b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-isopropyl-N-methylacetamide

Clear gum, 8.8mg (38%): Η NMR (400MHz, CDCI₃) δ 1.10 (3H,d), 1.22 (3H, m), 1.34 (3H, d), 2.39 (1 H, m), 2.80 (3H,s), 2.99-3.10 (4H, m), 3.17 (1 H, m), 3.73-3.99 (4H, m), 4.06-4.24 (4H, m), 7.31 (1 H, td), 7.43 (1H, td), 7.80 (2H, dd) LC-MS (A, 5.5min) Peak at time 3.33 minutes gave m/e 480 (M+H)⁺

Example 60

(3aS,6S,6aR)-4-[(1 R,6S)-bicyclo[4.1.0]hept-3-en-7-ylcarbonyl]-N,6-dimethyl-5- oxo-N-propylhexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

To a solution of (1R,6S)-bicyclo[4.1.0]hept-3-ene-7-carboxylic acid¹ (0.309g, 2.25mmol) in DCM (12ml) under nitrogen was added DMF (0.030ml) followed by oxalyl chloride (0.30ml, 3.42mmol). The solution was stirred at room temperature for one hour and then concentrated in vacuo. The residue was dissolved in toluene (5ml) and evaporated in vacuo twice. The resultant acid chloride was dried in vacuo.

To a cooled, -10°C, solution of intermediate 26 (0.413g, 1.5mmol) in THF (12ml) under nitrogen was added a solution of lithium hexamethyldisilazide in THF (1.65ml of 1.0M solution, 1.65mmol). After 15 minutes a solution of the acid chloride in THF (6ml) was added. The brown solution was stirred at -10°C for 2 hours. The reaction mixture was separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with brine and dried over anhydrous MgS0₄. The solvent was removed in vacuo to give a light brown gum. This was chromatographed over silica (Merck 9385) using cyclohexane/ethyl acetate (4:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.432g (73%).

Η NMR (CDCI₃): δ 5.50 (2H, m), 3.79-3.68 (2H, m), 3.44 (1H, dd, J=7,10Hz), 3.38 (1 H, m), 3.16 (2H, m), 3.01 (1 H, pent, J=7Hz), 2.85 (3H, s), 2.64 (1 H, m), 2.49-2.39 (2H, m), 2.42 (1 H, t, J=9Hz), 2.28-2.09 (2H, m), 1.94 (1 H, m), 1.70 (1H, m), 1.67-1.58 (3H, m), 1.27 (3H, d, J=7Hz), 0.93 (3H, t, J=7.5Hz). Mass spec, thermospray: +ve m/e: 396 [MH⁺] Ref. 1 H Musso and U. Bietham , Chem Ber, 1964, 97, 2282

Example 61

(3aS,6S,6aR)-4-[(1 R,5S)-bicyclo[3.1.0]hex-6-ylcarbonyl]-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

To a solution of (1 R,5S)-bicyclo[3.1.0]hex-6-ylcarboxylic acid¹ (0.248g, 2.0mmol) and triethylamine (0.279ml, 0.202g, 2.0ml) in DCM (10ml) at room temperature under nitrogen was added oxalyl chloride (0.174g, 0.254g, 2.0ml). After 30 minutes the solvent was removed under vacuum at room temperature and the residue was treated with diethyl ether (10ml). The solid was removed by filtration and washed with diethyl ether (2x5ml). The combined filtrate and washings were cooled to 0°C under nitrogen to give a 0.1 M solution of acid chloride. To a cooled, 0°C, solution of intermediate 26 (0.275g, I .Ommol) in THF (5ml) under nitrogen was added a solution of lithium hexamethyldisilazide in THF (1.1ml of 1.0M solution, 1.1 mmol). After 20 minutes the pale yellow solution was added dropwise to the cooled solution of acid chloride. The reaction mixture was stirred under nitrogen at 0°C for 20 minutes and then separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with water, saturated sodium bicarbonate solution and brine. The organic phase was iried over anhydrous MgS0₄. The solvent was removed in vacuo and chromatographed over silica (Merck 9385) using cyclohexane/ethyl acetate (2:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.298g (70%). Η NMR (CDCI₃): δ 5.78-5.54 (2H, m), 3.75-3.64 (2H, m), 3.48-3.32 (2H, m), 3.23-3.08 (2H, m), 2.99 (1 H, m), 2.90-2.37 (8H, m), 2.19-2.07 (1 H, m), 2.05-1.79 (1 H, m), 1.62 (2H, m), 1.25 (3H, m), 0.92 (3H, m). Mass spec, thermospray: +ve m/e: 382 [MH⁺] ¹Ref 1. J. Meinwald, S.S. Labana and M.S. Chadha J. Am. Chem. Soc, 85, 582-585 (1963) Example 62 and example 63

(3aS,6S,6aR)-4-(cyclohexylcarbonyl)-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-blpyrrole-1 (2H)-sulphonamide and (3aS,6S,6aR)-4- r(1 R,5S,6S)-bicyclo[3.1.0]hex-2-en-6-ylcarbonyl]-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b1pyrrole-1 (2H)-sulphonamide

Example 61 (0.038g, 0.1 mmol) was dissolved in ethyl acetate and hydrogenated over Pd/C (50% water, 0.017g) for 3 hours. The catalyst was removed by filtration and washed with ethyl acetate. The combined filtrate and washings were evaporated in vacuo and the residue was purified using preparative HPLC. This gave example 62 as a white solid, 0.018g (47%) and example 63 as a white solid, 0.006g (16%).

Example 62 Η NMR (CDCIg): δ 3.82-3.71 (2H, m), 3.47 (1 H, dd, J=6,10Hz), 3.42 (1 H, m),

3.17 (2H, m), 3.02 (1 H, m), 2.85 (3H, s), 2.68 (1 H, m), 2.12 (1 H, t, J=8Hz), 1.98 (1H, m), 1.94-1.82 (5H, m), 1.77 (1H, m), 1.68 (1 H, m), 1.63 (2H, m), 1.27 (3H, d, J=7.5Hz), 1.13 (1 H, m), 0.93 (3H, t, J=7.5Hz). Mass spec, thermospray: +ve m/e: 384 [MH⁺]

Example 63

Η NMR (CDCI₃): δ 3.81-3.69 (2H, m), 3.50-3.37 (2H, m), 3.28 (1 H, m), 3.17 (2H, m), 3.00 (1 H, m), 2.85 (3H, s), 2.69 (1 H, m), 2.01 (1 H, m), 1.90-1.67 (5H, m), 1.63 (2H, m), 1.47-1.18 (5H, m), 1.25 (3H, d, J=7.5Hz), 0.93 (3H, t, J=7.5Hz). Mass spec, thermospray: +ve m/e: 386 [MH⁺]

Example 64

(3aS,6S,6aR)-4-{[(1S,2S,3R,5R,6R)-2,3-dihydroxybicyclo[3.1.01hex-6- yl]carbonyl}-N,6-dimethyl-5-oxo-N-propyl-hexahydropyrrolo[3,2-blpyrrole-1(2H)- sulphonamide

To example 61 (0.148g, 0.39mmol) was added a solution of osmium tetroxide (0.11g, 0.43mmol) in THF (10ml). The reaction was stirred under nitrogen at room temperature for 48 hours. A solution of sodium metabisulphite (1g) in water (20ml) was added and the reaction mixture was stirred rapidly for 24 hours. DCM (50ml) was added and the phases were separated. The organic phase was dried over anhydrous MgS0₄ and evaporated in vacuo. The residue was purified using a Bond-elut cartridge and a cyclohexane/ethyl acetate gradient. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.093g (57%).

Η NMR (CDCI₃): δ 4.28 (1 H, m), 4.22-4.08 (1 H, m), 3.80-3.70 (2H, m), 3.50-3.42 (2H, m), 3.17 (2H, m), 3.03 (1H. m), 2.85 (3H, s), 2.70-2.50 (3H, m), 2.32 (1H, m), 2.23-1.87 (5H, m), 1.62 (2H, m), 1.27 (3H, d, J=7.5Hz), 0.93 (3H, t, J=7.5Hz).

Mass spec, thermospray: +ve m/e: 416 [MH⁺]

Example 65 (1 S,2S,3R,5R,6R)-6- (3S,3aR,6aS)-3-methyl-4-{fmethyl(propyl) aminolsulphonyl}-2-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)carbonyl]-2- {[(ethylamino)carbonyl]oxy}bicyclo[3.1.0]hex-3-yl ethylcarbamate

and

Example 66 (1 S,2S,3R,5R,6R)-6-f((3S,3aR,6aS)-3-methyl-4-{fmethyl(propyl) amino]sulphonyl}-2-oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)carbonyl]-2- {[(ethylamino)carbonyl]oxy}bicyclo[3.1.0]hex-3-yl ethylcarbamate:

To a solution of example 64 (0.018g, 0.043mmol) in DCM (2ml) was added triethylamine (0.015ml, 0.011g, 0.11 mmol), DMAP (cat.) and ethyl isocyanate (0.087ml, 0.078g, 1.1 mmol). The reaction mixture was left to stand for 12 days. The reaction mixture was applied to a preparative TLC plate and eluted with ethyl acetate/cyclohexane (3:1 v/v). The required bands were removed and eluted with ethyl acetate. The solvent was evaporated in vacuo to give Example 65 as a colourless gum, 0.009g (38%) and Example 66 as a colourless gum, 0.007g (29%). Example 65: NMR (CDCI₃): δ 5.35 (1 H, m), 5.09 (1 H, m), 4.64 (1 H, m), 4.54 (1 H, m), 3.82- 3.69 (2H, m), 3.51 (1 H, m), 3.42 (1H, m), 3.31 (2H, m), 3.27-3.10 (4H, m), 3.03 (1 H, m), 2.87 (3H, s), 2.68 (1 H, m), 2.54 (1 H, m), 2.28-1.94 (5H, m), 1.62 (2H, m), 1.28 (3H, d, J=7.5Hz), 1.12 (6H, m), 0.92 (3H, t, J=7.5Hz). Mass spec, thermospray: +ve m/e: 575 [MNH₄ ⁺]

Example 66:

Η NMR (CDCI₃): δ 5.36 (1 H, m), 5.01 (1 H, m), 4.67 (1 H, m), 4.55 (1 H, m), 3.83- 3.69 (2H, m), 3.49 (1 H, m), 3.41 (1 H, m), 3.27-3.09 (6H, m), 3.02 (1 H, m), 2.86 (3H, s), 2.68 (1 H, m), 2.60 (1 H, m), 2.27-1.94 (5H, m), 1.62 (2H, m), 1.28 (3H, d, J=7.5Hz), 1.12 (6H, m), 0.92 (3H, t, J=7.5Hz). Mass spec, thermospray: +ve m/e: 575 [MNH₄ ⁺]

Example 67

(1S,2S,3R,5R,6R)-6-[((3S,3aR,6aS)-3-methyl-4- {[methyl(propyl)amino]sulphonyl}-2-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)- yl)carbonyl]-2-(acetyloxy)bicyclo[3.1.0]hex-3-yl acetate

To a solution of example 64 (0.018g, 0.043mmol) in DCM (2ml) was added pyridine (0.036ml, 0.035g, 0.44mmol), DMAP (cat.) and acetic anhydride (0.042ml, 0.044g, 0.44mmol). The reaction mixture was left to stand for 3 days. The reaction mixture was purified using a Bond-elut cartridge and a cyclohexane/ethyl acetate gradient. The required fractions were combined and evaporated in vacuo to give the title compound as a colourless gum, 0.018g (84%). Η NMR (CDCI₃): δ 5.45 (1H, m), 5.21+5.12 (1H, m), 3.83-3.71 (2H, m), 3.53-

3.38 (2H, m), 3.18 (2H, m), 3.04 (1 H, m), 2.87 (3H, 2xs), 2.72-2.58 (2H, m), 2.22

(1 H, m), 2.17-1.96 (10H, m), 1.62 (2H, m), 1.28 (3H, d, J=7.5Hz), 0.93 (3H, t,

J=7.5Hz).

Mass spec, thermospray: +ve m/e: 500 [MH⁺]

Example 68

(3aS,6S,6aR)-4-[5-(hydroxymethyl)-1 ,3-thiazol-2-yl]-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrroie-1(2H)-sulphonamide Intermediate 27 (2.77g, 4.42mmol) was dissolved in THF (140ml) and treated with acetic acid (371 mg, 6.2mmol, 354ul) followed by TBAF (1M in THF, 5.3ml, 5.3mmol). The reaction mixture was stirred at room temperature for 2h 20min. The mixture was concentrated and the residue purified by chromatography on silica (Merck 7729) using ethyl acetate:cyclohexane 1 :4, 1 :1. 2:1 , then 4:1 as eluent to give the title compound (Rf 0.2 in 1 :1 ethyl acetate:cyclohexane) as a colourless foam(1.49g). nmr (CDCI₃): δ 7.3 (1H,s), 4.8 (2H,s), 4.11-4.02 (1H,m), 3.83 (1H,t), 3.65 (1 H,dd), 3.5 (1 H,m), 3.27-3.09 (3H,m), 2.88 (3H,s), 2.91-2.83 (1 H,m), 2.25 (1 H,m), 1.69-158 (3H,m), 1.28 (3H,d), 0.94 (3H,t)

LC/MS (method B): +ve m/e: 389 [MH]⁺ , RT = 2.94min

Example 69 (3aS,6S,6aR)-4-[5-(azidomethyl)-1 ,3-thiazol-2-yl]-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Example 68 (1.2g, 3.09mmol) was dissolved in DCM (25ml) and triethylamine (624mg, 6.18mmol, 860ul) added. The solution was cooled to 5°C. Methanesulphonyl chloride (708mg, 6.18mmol, 479ul) was added dropwise (temp =< 6°C). The cooling bath was removed and the reaction stirred for 2 h. Sodium azide (603mg, 9.28mmol) was added followed by DMF (25ml). The mixture was stirred at room temperature for 20h and then diluted with ethyl acetate (300ml) and the organic solution washed with water, brine, and dried (MgS04). The organic extract was concentrated and the residue purified using a 10g SPE silica cartridge eluting with DCM (x2), chloroform (x2), diethyl ether (x2) and ethyl acetate (x2) to give the title compound (Rf 0.6 in 1 :1 ethyl acetate:cyclohexane) as a white solid (1.02g). nmr (CDCI₃): δ 7.35 (1 H,s), 4.46 (2H,s), 4.02 (1 H.m), 3.85 (1 H,t), 3.7 (1 H,dd), 3.52 (1 H,m), 3.28-3.11 (3H,m), 2.88 (3H,s), 2.92-2.84 (1 H,m), 2.28 (1H,m), 1.64 (2H,m), 1.3 (3H,d), 0.94 (3H,t) LC/MS (method B): +ve m/e: 414 [MH]⁺ , RT = 3.54min

Example 70 tert-butyl [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yi]methylcarbamate

Example 69 (1.01g, 2.45mmol) and Boc₂0 (3.2g) in DCM (50ml) and iso- propanol (50ml) was hrdrogenated over 10% palladium-on-carbon (500mg) for 3.5h. The catalyst was removed by filtration and the filtrate concentrated to give a colourless oil. The crude product was purified by chromatography on silica (Merck 7729) using ethyl acetate:cyclohexane 1 :1 as eluent to give the title compound (Rf 0.32 in 1 :1 ethyl acetate:cyclohexane) as a white solid (661 mg). Η nmr (CDCl₃): δ 7.25 (1 H,s), 4.84 (1 H, br s), 4.5-4.35 (2H,m), 4.1-4.0 (1 H,m), 3.83 (1 H,t), 3.67 (1 H,dd), 3.5 (1 H,m), 3.3-3.1 (3H,m), 2.88 (3H,s), 2.9-2.8 (1 H,m), 2.25 (1 H,m), 1.7-1.6 (2H,m), 1.45 (9H,s), 1.28 (3H,d), 0.94 (3H,t) LC/MS (method B): +ve m/e: 488 [MH]⁺ , RT = 3.43min

Example 71

(3aS,6S,6aR)-4-[5-(aminomethyl)-1 ,3-thiazol-2-yl]-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Example 70 (230mg, 0.47mmol) was dissolved in DCM (10ml) and treated with trifluoroacetic acid (4ml). After standing at room temperature for 30min the reaction mixture was concentrated and the residue azeotroped with heptane. The trifluoroacetate salt was dissolved in DCM and the solution washed with saturated aqueous sodium bicarbonate. The organic layer was separated, dried (MgS0₄) and concentrated to give the title compound as a colourless gum (169mg)

Η nmr (CDCI₃): δ 7.21 (1 H,s), 4.12-4.03 (1 H,m), 4.02 (2H,s), 3.83 (1 H,t), 3.68 (1 H,dd), 3.5 (1 H,m), 3.27-3.09 (3H,m), 2.88 (3H,s), 2.91-2.82 (1 H,m), 2.25 (1H,m), 1.69-1.53 (4H,m), 1.28 (3H,d), 0.94 (3H,t) LC/MS (method B): +ve m/e: 388 [MH]⁺ , RT = 2.27min

Example 72 N-{[2-((3S,3aR,6aS)-3-methyl-4-{[methyi(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yl]methyl}isoxazole-5- carboxamide

lsoxazole-5-carboxylic acid (11.6mg, 0.1 mmol) was dissolved in acetonitrile

(0.3ml) and DIPEA (27mg, 0.2mmol, 36ul) added. EDC (19.7mg. 0.1 mmol) was added followed by a solution of HOBT (7.7mg, 0.057mmol) in acetonitrile (0.2ml). Example 71 (20.1 mg, 0.052mmol) in acetonitrile (0.5ml) was added and the reaction allowed to stand at room temperature for 3 days. The reaction mixture was concentrated and the residue dissolved in DCM (5ml). The DCM solution was washed with 1 N aqueous hydrochloric acid, then saturated sodium bicarbonate solution, and concentrated to give a brown gum. The crude product was purified using a 2g SPE silica cartridge eluting with DCM (x2), diethyl ether (x2) and ethyl acetate (x2) to give the title compound (Rf 0.7 in ethyl acetate) as a colourless gum (7mg).

Η nmr (CDCI₃): δ 8.26 (1 H,s), 7.19 (1 H,s), 6.89 (2H,br s), 4.7 (2H,m), 4.04-3.95 (1 H,m), 3.76 (1 H,t), 3.6 (1 H,dd), 3.44 (1 H,m), 3.2-3.02 (3H,m), 2.81 (3H,s), 2.84- 2.75 (1H,m), 2.18 (1H,m), 1.64-1.5 (2H,m), 1.2 (3H,d), 0.83 (3H,t) LC/MS (method B): +ve m/e: 483 [MH]⁺ , RT = 3.06min

Example 73

(3aS,6S,6aR)-N,6-dimethyl-4-(5-{[(methylsulphonyl) amino]methyl}-1 ,3-thiazol-2- yl)-5-oxo-N-propylhexahydropyrrolo[3,2-b]pyrroie-1 (2H)-sulphonamide

Example 71 (20.1 mg, 0.052mmol) was dissolved in DCM (1 ml) and treated with DIPEA (18.1 ul, O.l mmol) followed by methanesulphonyl chloride (8ul, O.lmmol). The reaction mixture was allowed to stir at room temperature for three days. The mixture was diluted with DCM (5ml), washed with aqueous hydrochloric acid, (1 N, 5ml), saturated sodium bicarbonate solution (5ml), and concentrated to give the title compound (Rf 0.59 in ethyl acetate) as a white solid (15mg) Η nmr (CDCI₃): δ 7.83 (1 H,s), 4.93 (1 H,t), 4.50 (2H,d), 4.07 (1 H,m), 3.83 (1 H,t), 3.67 (1 H,dd), 3.5 (1 H,m), 3.28-3.08 (3H,m), 2.92 (3H,s), 2.88 (3H,s), 2.91-2.82 (1 H,m), 2.26 (1 H,m), 1.65 (2H,m), 1.28 (3H,d), 0.94 (3H,t) LC/MS (method B): +ve m/e: 466 [MH]⁺ , RT = 2.98min Example 74

(3aS,6S,6aR)-4-[5-({[(ethylamino)carbonyl] amino}methyl)-1 ,3-thiazol-2-yl]-N,6- dimethyl-5-oxo-N-propylhexahydroρyrrolo [3,2-b]pyrrole-1 (2H)-sulphonamide

Example 71 (20.1 mg, 0.052mmol) was dissolved in DCM (1 ml) and treated with DIPEA (9ul, 0.052mmol) followed by ethyl isocyanate (8.2ul, O.lmmol). The reaction mixture was allowed to stir at room temperature for three days. The solvent was removed in vacuo and the residue purified using a 2g SPE silica cartridge eluting with DCM (x2), diethyl ether (x2) and ethyl acetate (x2) 4:1 ethyl acetate:methanol (x1 ) to give the title compound as a white solid (21 mg) nmr (CDCI₃): δ 7.24 (1 H,s), 4.89 (1 H,t), 4.55 (1 H.t), 4.48 (2H,d), 4.04 (1 H,m), 3.81 (1 H,t), 3.61 (1 H,dd), 3.49 (1 H,m), 3.3-3.01 (5H,m), 2.87 (3H,s), 2.91-2.78 (1 H,m), 2.21 (1 H,m), 1.64 (2H,m), 1.26 (3H,d), 1.12 (3H,t), 0.94 (3H,t)

LC/MS (method B): +ve m/e: 459 [MH]⁺ , RT = 2.92min

Example 75 [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl) amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yl]methyl carbamate

Example 68 (29mg, 0.075mmol) was dissolved in DCM (2ml) and treated with chloroacetylisocyanate (26.8mg, 0.22mmol, 19ul). After 1h methanol (1ml) and triethylamine (104ul, 0.75mmol) was added and the reaction stirred for 1h. The reaction mixture was concentrated in vacuo and the residue dissolved in DCM:methanol (3:2, 4ml) and treated with triethyamine (104ul). After 7h the mixture was concentrated and the residue purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 2:1 ethyl acetate:cyclohexane as eluent to give the title compound (Rf 0.4 in 2:1 ethyl acetate:cyclohexane) as a white solid (18mg) nmr (CDCI₃): δ 7.19 (1 H,s), 5.14 (2H, s), 4.59 (2H,br), 4.1-3.92 (1H,m), 3.76 (1H,t), 3.61 (1 H,dd), 3.44 (1 H,m), 3.24-3.0 (3H,m), 2.88-2.72 (4H,m), 2.18 (1H,m), 1.66-1.44 (2H,m), 1.21 (3H,d), 0.87 (3H,t) LC/MS (method A): +ve m/e: 432 [MH]⁺ , RT = 4.11 min Example 76

[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl) amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yl]methyl cyclopropylcarbamate

Cyclopropylamine (500mg, 8.76mmol, 606ul) and pyridine (2.77g, 35.04mmol, 2.83ml) were dissolved in DCM (20ml) and cooled to 0°C. A solution of phosgene (1.93M in toluene, 5.9ml, 11.4mmol) was added rapidly and the mixture allowed to stir at 0°C for 2h. Dilute hydrochloric acid (0.5N, 20ml) was added and the mixture separated througn a hydrophobic frit. A portion of the above solution (7ml) was added to Example 68 (31 mg, O.Oδmmol) and the mixture allowed to stand at room temperature for 4 days. The mixture was concentrated and the residue purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :1 ethyl acetate:cyclohexane as eluent to give the title compound (Rf 0.32 in 1 :1 ethyl acetate:cyclohexane) as a white solid (28mg) Η nmr (CDCI₃): δ 7.38 (1 H,s), 5.2 (2H,br s), 4.95 (1 H,br s), 4.1-4.0 (1 H,m), 3.83 (1 H,t), 3.67 (1 H,dd), 3.5 (1 H,m), 3.27-3.08 (3H,m), 2.92-2.8 (4H,m), 2.57

(1 H,br), 2.25 (1 H,m), 1.63 (2H,m), 1.27 (3H,d), 0.94 (3H,t), 0.71 (2H,br), 0.51

(2H, br)

LC/MS (method B): +ve m/e: 472 [MH]⁺ , RT = 3.24min

Example 77

[2-((3S,3aR,6aS)-3-methyl-4-{ methyl(propyl) amino1sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-thiazol-5-yQmethyl ethylcarbamate

Translactam alcohol example 68 (75 mg, 0.19mmol) was dissolved in dry dichloromethane (2 mL). Triethylamine (29.6 μL, 0.21 mmol, 1.1 equiv) was added to the solution followed by ethyl isocyanate (76.5 μL, 0.96 mmol, 5 equiv). The mixture was left to stand at room temperature for 90 hours before it was purified using a SPE 2g silica cartridge eluted with dichloromethane, chloroform, cyclohexane, 1 :1 ether: cyclohexane, 2:1 ether: cyclohexane, 3:1 ether: cyclohexane, ether, 2:1 cyclohexane: ethyl acetate, 1 :1 cyclohexane: ethyl acetate to give the title compound as a white solid, 77mg, 86%. LC/MS (A, 5.5min) Peak at 3.31 min gives m/e 460 [MH]\

Example 78 tert-butyl [[((3aS,6S,6aR)-4-[5-(hydroxymethyl)-1 ,3-thiazol-2-yl]-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl] (methyl)amino]acetate

Intermediate 30 (374mg, 0.53mmol) was dissolved in dry tetrahydrofuran (11ml), glacial acetic acid (40μl, 0.69mmol) then tetrabutyl ammonium fluoride (580μl, 0.58mmol, 1.0M solution in tetrahydrofuran) were added and the mixture stirred under N₂ at room temperature for 5 hours. The solvent was removed and the residual syrup azeotroped with toluene (10ml) and dried in vacuo to give a white solid. This was taken up in chloroform and purified using solid phase extraction, Bond-Elut silica column 10g, eluted as follows: chloroform, diethyl etheπcyclohexane 2:1 , diethyl ether, diethyl etheπcyclohexane 1 :1 , ethyl acetate yclohexane 1 :1 , ethyl acetate. Appropriate fractions were combined, evaporated under reduced pressure and dried in vacuo to give the title compound as a white solid 139mg (56%): Η NMR (400MHz, CDCI₃) δ 1.29 (3H, s), 1.48 (9H, s), 1.76 (1H, s), 2.27 (1H, m), 2.87 (1H, m), 3.03 (3H, s), 3.14 (1 H, m), 3.64-3.94 (5H, m), 4.07 (1 H, m), 4.82 (2H, s), 7.32 (1 H, s) LC-MS (A, 5.5min) Peak at time 3.06 minutes gives m/e 461 (M+H)⁺

Example 79

[2-((3S,3aR,6aS)-4-{[(2-{[(1 S)-2-hydroxy-1-phenylethyl]amino}-2- oxoethyl)(methyl)amino]sulphonyl}-3-methyl-2-oxo hexahydropyrrolo[3,2- b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yl]methyl ethylcarbamate Intermediate 32 (17.4mg, 0.037mmol) was dissolved in acetonitrile (1 ml) and di- isopropyl ethylamine (14.2ul, 0.081 mmol), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (7.8mg, 0.041 mmol) and hydroxybenztriazole (7.5mg, 0.056mmol) and (S)-2-phenylaminoethanol (5.5mg, 0.041 mmol) were added and the mixture left for 4 days. The acetonitrile was removed using a stream of nitrogen and the residue taken up in dichloromethane (1ml), washed with 0.1 M aqueous hydrochloric acid (500ul), separated then washed with saturated sodium hydrogen carbonate (1 ml) and then separated using a hydrophobic frit. The collected organic phase was concentrated and then purified using preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 5% v/v methanol in ethyl acetate eluent to give the title compound as a white solid 14.9mg (68%): Η NMR (250MHz, CDCI₃) δ 1.14 (3H, t), 1.26 (3H, d), 2.13-2.36 (2H, m), 2.89 (1 H, m), 3.03 (3H, s), 3.07- 3.30 (3H, m), 3.50-3.63 (1 H, m), 3.72 (1H, dd), 3.80-4.18 (6H, m), 5.06-5.16 (1H, m), 5.20 (2H, s), 6.95 (1 H, d), 7.28-7.42 (6H, m) LC-MS (A, 5.5min) Peak at time 2.93 minutes gives m/e = 595 (M+H)⁺

Example 80

[2-((3S,3aR,6aS)-4-{[(2-{[(3-methoxyisothiazol-5-yl)methyl]amino}-2- oxoethyl)(methyl)amino]sulphonyl}-3-methyl-2-oxohexahydro pyrrolo[3,2- b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-5-yl]methyl ethylcarbamate

Intermediate 32 (23mg, 0.048mmol) was dissolved in DCM (1 ml). Acetonitrile (0.5ml) and DMF (0.5ml) were added followed by DIPEA (42ul, 0.24mmol). EDC (11mg, 0.058mmol) was added followed by 3-methoxy-5- aminomethylisothiazole¹ (10.5mg, 0.058mmol) and HOBT (6.5mg, 0.048mmol). The reaction was stirred at room temperature for 18h. The solvents were removed in vacuo and the residue dissolved in ethyl acetate. The ethyl acetate solution was washed with 1 N aqueous hydrochloric acid, saturated sodium bicarbonate solution, dried (MgS0₄) and concentrated to give a brown gum. The crude product was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using ethyl acetate as eluent to give the title compound (Rf 0.61 in ethyl acetate) (3mg)

LC/MS (method B): +ve m/e: 602 [MH]⁺ , RT = 2.9min

1. Lykkeberg, Jytte; Krogsgaard-Larsen, Povl. Acta Chem. Scand. Ser B (1976), B30(8). 781-5

Example 81

(3aS,6S,6aR)-4-f4-(hydroxymethyl)-1 ,3-thiazol-2-yl1-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

To a solution of intermediate 33 (0.463g, 0.74mmol) in THF (15ml) was added HF/pyridine (1.8ml) and the solution was stirred at room temperature for 3 hours. The reaction mixture was poured into ethyl acetate and saturated sodium bicarbonate solution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organics were washed with 0.5N hydrochloric acid and brine. The organic phase was dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (1 :1 v/v) as eluant to give the title compound as a white solid, 0.20g (70%).

Η NMR (CDCI₃): δ 6.87 (1 H, s), 4.66 (2H, d, J=6Hz), 4.10 (1 H, m), 3.84 (1 H, m), 3.68 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.30-3.08 (3H, m), 2.90 (1H, m), 2.88 (3H, s), 2.28 (1 H, m), 2.01 (1 H, t, J=6Hz), 1.64 (2H, hextet, J=7.5Hz), 1.29 (3H, d, J=7.5Hz), 0.94 (3H, t, J=7.5Hz).

Mass spec, thermospray: +ve m/e: 389 [MH⁺]

Example 82 (3aS,6S,6aR)-4-(4-formyl-1 ,3-thiazol-2-yl)-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Example 81 (30.8mg, 0.79mmol) was dissolved in chloroform (2ml). Manganese (IV) oxide (300mg, 3.45mmol) was added and the mixture stirred for 4 hours. The mixture was filtered and the filter cake washed with 10% v/v methanol in chloroform (30ml). The filtrate was evaporated to give the title compound as a white solid 23.2mg (76%):

Η NMR (400MHz, CDCI₃) δ 0.95 (3H, t), 1.32 (3H,d), 1.65 (2H,m), 2.32 (1 H, m), 2.99 (3H, s), 3.00 (1 H, m),

3.12-3.29 (3H, m), 3.54 (1 H, td), 3.72 (1 H, dd), 3.87 (1 H, t), 4.15 (1 H, m), 7.89

(1H, s), 9.90 (1H, s)

LC-MS (A, δ.δmin)

Peak at time 3.11 min gives m/e = 387 (M+H)⁺

Example 83

[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-thiazol-4-yi]methyl ethylcarbamate

To a solution of example 81 (0.020g, O.Oδmmol) in dichloromethane (2ml) was added triethylamine (0.007ml, δ.Omg, O.Oδmmol) and ethyl isocyanate (0.041 mi, 37mg, O.δ2mmol). The colourless solution was left to stand for 13 days. The solvent was removed in vacuo and the residue was purified by preparative HPLC to give the title compound as a white solid, 0.017g (74%).

Η NMR (CDCIg): δ 6.93 (1 H, s), δ.09 (2H, m), 4.71 (1 H, broad s), 4.09 (1 H, m), 3.83 (1H, m), 3.68 (1H, dd, J=6, 10Hz), 3.51 (1H, m), 3.32-3.07 (5H, m), 2.91 (1H, m), 2.88 (3H, s), 2.26 (1 H, m), 1.64 (2H, hextet, J=7Hz), 1.28 (3H, d, J=7.5Hz), 1.16 (3H, t, J=7Hz), 0.94 (3H, t, J=7Hz). Mass spec, thermospray: +ve m/e: 460 [MH⁺]

Example 84

[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolof3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]methyl carbamate

To a solution of example 81 (0.020g, O.Oδmmol) in dichloromethane (2ml) was added chloroacetylisocyanate (0.013ml, 0.018g, O.l δmmol). The reaction mixture was left to stand for 90 minutes and then methanol (2ml) and δ triethylamine (0.060ml, 0.040g, 0.4mmol) were added. After 60 minutes the solvent was removed in vacuo and the residue was purified by preparative

HPLC to give the title compound as a white solid, 0.01 δg (70%)

Η NMR (CDCI₃): δ 6.97 (1 H, s), 5.10 (2H, m), 4.68 (1 H, broad s), 4.10 (1 H, m),

3.83 (1 H, m), 3.68 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.30-3.08 (3H, m), 2.92

(1H, m), 2.88 (3H, s), 2.26 (1 H, m), 1.64 (2H, hextet, J=7Hz), 1.28 (3H, d,

J=7.5Hz), 0.94 (3H, t, J=7Hz).

Mass spec, thermospray: +ve m/e: 432 [MH⁺]

Example 8δ

(3aS,6S,6aR)-4-[4-(2-hydroxyethyl)-1 ,3-thiazol-2-yl]-N,6-dimethyl-δ-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

To a solution of intermediate 34 (0.390g, 0.61 mmol) in THF (15ml) was added HF/pyridine (1.5ml) and the solution was stirred at room temperature for 4 hours. The reaction mixture was poured into ethyl acetate and saturated sodium bicarbonate solution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organics were washed with 0.5N hydrochloric acid and brine. The organic phase was dried over anhydrous magnesium sulphate and evaporated in vacuo. The residue was chromatographed over silica (Merck 9385) using medium pressure (~4psi) and cyclohexane/ethyl acetate (1 :1 v/v) as eluant to give the title compound as a white solid, 0.182g (74%).

Η NMR (CDCI₃): δ 6.67 (1 H, s), 4.10 (1 H, m), 3.90 (2H, q, J=6Hz), 3.85 (1H, m), 3.69 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.41 (1 H, m), 3.30-3.08 (3H, m), 2.89 (2H, t, J=6Hz), 2.87 (3H, s), 2.86 (1 H, m), 2.31 (1 H, m), 1.64 (2H, hextet, J=7Hz), 1.27 (3H, d, J=7.5Hz), 0.94 (3H, t, J=7Hz). Mass spec, thermospray: +ve m/e: 403 [MH⁺]

Example 86

2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyi(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]ethyl allylcarbamate To a solution of example 85 (0.022g, 0.0δ4mmol) in dichloromethane (2ml) was added triethylamine (0.007δml, δ.δmg, 0.0δ4mmol) and allyl isocyanate (0.019ml, 18mg, 0.22mmol). The colourless solution was left to stand for 13 days. The solvent was removed in vacuo and the residue was purified by δ preparative HPLC to give the title compound as a white solid, 0.024g (92%). Η NMR (CDCI₃): δ 6.66 (1 H, s), δ.83 (1 H, m), 5.17 (1 H, m), 5.12 (1H, m), 4.69 (1H, broad s), 4.38 (2H, t, J=7Hz), 4.08 (1H, m), 3.82 (1H, m), 3.78 (2H, m), 3.67 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.30-3.06 (3H, m), 2.97 (2H, t, J=7Hz), 2.91 (1 H, m), 2.88 (3H, s), 2.25 (1 H, m), 1.64 (2H, hextet, J=7Hz), 1.28 (3H, d, 0 J=7.δHz), 0.94 (3H, t, J=7Hz).

Mass spec, thermospray: +ve m/e: 486 [MH⁺]

Example 87 δ Similarly prepared to example 86

2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]ethyl propylcarbamate

0 White solid, 0.018g (68%)

Η NMR (CDCI₃): δ 6.6δ (1 H, s), 4.62 (1 H, broad s), 4.37 (2H, t, J=7Hz), 4.08 (1H, m), 3.82 (1 H, m), 3.67 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.30-3.06 (5H, m), 3.00-2.84 (3H, m), 2.88 (3H, s), 2.25 (1 H, m), 1.64 (2H, hextet, J=7Hz), 1.50 (2H, m), 1.28 (3H, d, J=7.5Hz), 0.94 (3H, t, J=7Hz), 0.91 (3H, t, J=7Hz). δ Mass spec, thermospray: +ve m/e: 488 [MH⁺]

Example 88

Similarly prepared to example 86 0 2-f2-((3S,3aR,6aS)-3-methyl-4-{rmethyl(propyl)aminolsulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]ethyl 2-thien-2- ylethylcarbamate

White solid, 0.02δg (83%) NMR (CDCI₃): δ 7.17 (1 H, dd,J=1 ,δHz), 6.93 (1H, dd, J=3,δHz), 6.81 (1H, m),

6.63 (1 H, s), 4.77 (1 H, m), 4.38 (2H, t, J=7Hz), 4.08 (1 H, m), 3.82 (1 H, m), 3.67

(1H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.44 (2H, m), 3.30-2.84 (8H, m), 2.88 (3H, s),

2.24 (1H, m), 1.63 (2H, hextet, J=7.5Hz), 1.28 (3H, d, J=7.δHz), 0.94 (3H, t,

J=7Hz).

Mass spec, thermospray: +ve m/e: δδ6 [MH⁺]

Example 89 Similarly prepared to example 86 ethyl 3-[({2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4- yl]ethoxy}carbonyl)amino]propanoate

White solid, 0.025g (85%)

Η NMR (CDCI₃): δ 6.65 (1 H, s), 5.16 (1 H, broad s), 4.36 (2H, t, J=7Hz), 4.1 δ (2H, q, J=7Hz), 4.08 (1 H, m), 3.82 (1 H, m), 3.67 (1 H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.43 (2H, m), 3.30-3.07 (3H, m), 3.00-2.84 (3H, m), 2.88 (3H, s), 2.52 (2H, m), 2.25 (1 H, m), 1.63 (2H, hextet, J=7.5Hz), 1.27 (6H, m), 0.94 (3H, t, 0 J=7.δHz).

Mass spec, thermospray: +ve m/e: 646 [MH⁺]

Example 90 δ Similarly prepared to example 86

2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]ethyl isopropylcarbamate

0 White solid, 0.017g (65%) NMR (CDCI₃): δ 6.66 (1 H, s), 4.43 (1 H, m), 4.36 (2H, t, J=7Hz), 4.08 (1 H, m), 3.82 (1 H, m), 3.78 (1 H, m), 3.67 (1H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.30-3.07 (3H, m), 3.00-2.84 (3H, m), 2.88 (3H, s), 2.26 (1 H, m), 1.64 (2H, hextet, J=7.5Hz), 1.28 (3H, d, J=7.5Hz), 1.13 (6H, d, J=6Hz), 0.94 (3H, t, J=7Hz). δ Mass spec, thermospray: +ve m/e: 488 [MH⁺] Example 91

Similarly prepared to example 86 2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-thiazol-4-yl]ethyl benzylcarbamate

Pale pink solid, 0.019g (66%) NMR (CDCI₃): δ 7.38-7.22 (5H, m), 6.64 (1 H, s), 4.94 (1 H, m), 4.42 (2H, t,

J=7Hz), 4.3δ (2H, m), 4.07 (1 H, m), 3.81 (1 H, m), 3.6δ (1H, dd, J=6, 10Hz), 3.49 (1 H, m), 3.30-3.06 (3H, m), 3.01-2.82 (3H, m), 2.88 (3H, s), 2.23 (1 H, m), 1.64 (2H, hextet, J=7.δHz), 1.28 (3H, d, J=7.δHz), 0.94 (3H, t, J=7.δHz). Mass spec, thermospray: +ve m/e: 636 [MH⁺]

Example 92

Similarly prepared to example 86

2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4-yl]ethyl phenylcarbamate

White solid, 0.024g (85%) NMR (CDCI₃): δ 7.40-7.24 (5H, m), 7.06 (1 H,s), 6.88 (1 H, m), 6.74 (1 H, s), 4.48 (2H, t, J=6.5Hz), 4.19 (1 H, m), 3.78 (1H, m), 3.66 (1H, dd, J=6, 10Hz), 3.51 (1 H, m), 3.28-2.94 (6H, m), 2.87 (3H, s), 2.18 (1 H, m), 1.63 (2H, hextet, J=7.5Hz), 1.28 (3H, d, J=7.δHz), 0.93 (3H, t, J=7.δHz). Mass spec, thermospray: +ve m/e: 622 [MH⁺]

Example 93

Similarly prepared to example 86 ethyl [({2-[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl) amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-4- yl]ethoxy}carbonyl)amino]acetate White solid, 0.020g (71%)

Η NMR (CDCI₃): δ 6.67 (1H, s), 5.13 (1H, m), 4.39 (2H, t, J=7Hz), 4.22 (2H, q, J=7Hz), 4.06 (1 H, m), 3.94 (2H, d, J=5Hz), 3.82 (1 H, m), 3.67 (1H, dd, J=6, 10H-Z), 3.51 (1 H, m), 3.30-3.07 (3H, m), 3.02-2.84 (3H, m), 2.88 (3H, s), 2.26 (1 H, m), 1.64 (2H, hextet, J=7.5Hz), 1.28 (6H, m), 0.94 (3H, t, J=7.δHz). Mass spec, thermospray: +ve m/e: 619 [MH⁺]

Example 94

(3aS,6S,6aR)-4-[6-(hydroxymethyl)-1 ,3-benzothiazol-2-yl]-N,6-dimethyl-δ-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

To the trans-lactam intermediate 35, (0.495g,0.73mol, 1eq.) in dry THF(3 mL) was added pyridinium poly (hydrogen fluoride) (0.5 ml). The reaction mixture was stirred for 2.7δ hours and was then cooled (ice/water bath) whilst carefully basifying with 5% aqueous sodium carbonate solution. The solution was then extracted with ethyl acetate (X3) and the combined organic extracts were dried

((MgS0₄ ) and the product was purified by flash column chromatography over silica (Merck 9385 silica gel, 100 mL) eluting with cyclohexane :ethyl acetate

(3:2 then 1 :1 ). The title compound was obtained as a white solid (0.285g, 89%).

Mass Spec. : 439 [MH]⁺.

Η nmr (CDCQ : δ 7.83 (1 H,s,7'-H), 7.77 (1 H, d,4'-H), 7.43 (1H,d, δ'-H),4.81

(2H,d,arCH₂), 4.26-4.10 (1 H,m,δ-H), 3.88 (1 H,t,6a-H), 3.71 (1 H.dd.5-H or 3a-H), 3.62-3.48 (1 H,m, 5-H or 3a-H), 3.32-2.98 (4H,m,3-H,6-H, CH₂N), 2.89 (3H.S.N-

Me), 2.44-2.22 (m, 6-H), 1.80 (1H, t, OH),1.72-1.57 (2H,m, MeCH₂), 1.31

(3H,d,CH3). 0.96 (3H,t,CH₃) ppm.

HPLC: 12.1 min. (2.4%), 12.7 min. (1%), 26.9 min. (95%).

LCMS: 3.17 min. 439 [MH]⁺.

Example 95 tert-butyl [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulphonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-benzothiazol-6- yljmethylcarbamate To intermediate 36 (31.6 mg,4δμmol, 1eq.) in THF (3mL), was added acetic acid (3.3μL,δ9μmol,1.3 eq.) followed by a 1.0M solution of TBAF in THF (50μl, δ0μmol,1.1eq.). The reaction mixture was stirred at room temperature δ overnight. Due to incomplete reaction more acetic acid (6.6μL,118μmol, 2.6 eq.) and 1.0M TBAF in THF (100μl,100μmol, 2.2eq.) were added and the reaction mixture was stirred over the weekend. The reaction mixture was diluted with ethyl acetate (10 mL) and washed with sat. sodium chloride solution (10 mL). The organic phase was dried (MgS0₄) and the solvent was evaporated in vacuo. 0 The title compound was purified by preparative tic, eluting with cyclohexane:ethyl acetate (7:3)and was obtained as a white solid (6.8 mg,

28%).

Η nmr (CDCi₃) : δ 7.73 (2H,m,arH), 7.34 (1 H, d, arH), 4.91 (1 H, s.lMH). 4.42

(2H,d,arCH₂), 4.26-4.11 (1 H.rn.5-H), 3.87 (1 H,t,6a-H), 3.72 (1 H.dd.5-H or 3a-H), 5 3.62-3.42 (1 H.m, δ-H or 3a-H), 3.33-2.97 (4H,m,3-H,6-H,CH₂N), 2.89

(3H,s,NMe), 2.44-2.23 (1 H,m,6-H), 1.74-1.66 (2H,m,CH₂), 1.47 (9H,s,t-Bu), 1.32

(3H,d,CH₃), 0.9δ (3H,t, CH₃) ppm.

Mass Spec. : 638 [MH]⁺, 438 [MH-isobutylene-C0₂]⁺.

LCMS: 3.67 min. 638 [MH]⁺. 0

Example 96

(3aS,6S,6aR)-N-allyl-4-[δ-(hydroxymethyl)-1 ,3-thiazol-2-yl]-N,6-dimethyl-δ- oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide δ

Intermediate 44 (346mg, 0.55 mmol) was dissolved in dry tetrahydrofuran (11 mL) under nitrogen. Glacial acetic acid (41.21 μL, 1.3 equiv.) was added to the stirred mixture followed by tetra-n-butyl. ammonium fluoride (0.61 mL, 1.1 equiv., 1.0 M in tetrahydrofuran). The mixture was stirred at room temperature for 2 hours before 30 the solvent was removed under reduced pressure. The residue was azeotroped with toluene. The crude product was purified using a SPE 10g, silica cartridge eluted with dichloromethane, chloroform, 2:1 ether: cyclohexane, ether, 1 :1 cyclohexane: ethyl acetate, ethyl acetate (χ3) to give the title compound as a white solid, 203 mg (94%): 116

Η nmr (CDCI₃): δ 7.31 (1H, s), δ.83 (1 H, m), δ.30 (1H, m), δ.27 (1H, m), 4.81 (2H, bs), 4.08 (1 H, m), 3.86 (2H, m), 3.79 (1 H, td), 3.68 (1 H, dd), 3.63 (1 H, m), 3.14 (1H, m), 2.89 (1 H, m), 2.8δ(3H, s), 2.26 (1 H, m), 1.86 (1 H, bs), 1.29 (3H, d): LC/Mass spec, electrospray: +ve m/e: 387 [MH]⁺:

Example 97

[2-((3S,3aR,6aS)-4-{[allyl(methyl)amino]sulphonyl}-3-methyl-2- oxohexahydropyrro!o[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-δ-yl]methyl ethylcarbamate

Example 96 (190 mg, 0.49 mmol) was dissolved in dry dichloromethane (10 mL). Triethylamine (74.7 μL, 0.54 mmol, 1.1 equiv) was added to the solution followed by ethyl isocyanate (424 μL, 6.36 mmol, 10.9 equiv). The mixture, a solution, was left to stand at room tempreature for 97 hours before the solvent was removed under reduced pressure. The residue was purified using a SPE δg, silica cartridge eluted with dichloromethane, chloroform, cyclohexane, 1 :1 ether: cyclohexane, 3:1 ether: cyclohexane, ether, 2:1 cyclohexane: ethyl acetate, 1 :1 cyclohexane: ethyl acetate, ethyl acetate (χ2) to give the title compound as a white foam, 223 mg (99%), became a solid on trituration with ether: Η nmr (CDCI₃): δ 7.38 (1 H, s), δ.82 (1 H, m), δ.30 (1 H, m), 6.27 (1 H, m), 6.20 (2H, s), 4.68 (1 H, bm), 4.08 (1 H, m), 3.87 (2H, m), 3.79 (1 H, td), 3.68 (1 H, dd), 3.63 (1H, m), 3.23 (2H, m), 3.15 (1 H, m), 2.89 (1H, m), 2.8δ(3H, s), 2.26 (1 H, m), 1.28 (3H, d), 1.14 (3H, t): LC/Mass spec, electrospray: +ve m/e: 468 [MH]⁺:

Example 98

[2-((3S,3aR,6aS)-4-{[(2,3-dihydroxypropyl)(methyl) amino]sulphonyl}-3-methyl-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-δ-yl]methyl ethylcarbamate

Example 97 (71.3 mg, 0.16 mmol) was dissolved in tetrahydrofuran (2 mL). A solution of osmium tetroxide (44 mg, 0.17 mmol, _.1.1 equiv) in tetrahydrofuran (1.1 mL) was added to the stirred solution at room temperature. The mixture rapidly changed colour from yellow to brown, to black. The mixture was stirred for 22 hours before saturated aqueous sodium sulphite solution (2 mL) was added and the mixture stirred a further 2 hours 20 minutes. Then the mixture was diluted with ethyl acetate (20 mL) and washed with saturated aqueous sodium sulphite solution (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL). The combined organic phase was dried (MgS0₄), and evaporated to leave a black gum. The gum δ was purified using a SPE 1g, silica cartridge eluted with dichloromethane, chloroform, 2:1 ether: cyclohexane, ether, 1 :1 cyclohexane: ethyl acetate, ethyl acetate (χ3), 1:10 methanol: ethyl acetate to give the title compound as a white solid, 30 mg (39%): nmr (CDCI₃): δ 7.38 (1 H, s), 6.20 (2H, s), 4.70 (1 H, bm), 4.08 (1 H,td), 3.94 (1 H, 0 m), 3.86 (1 H, t), 3.65 (4H, m), 3.36 (2H, m), 3.20 (3H, m), 3.01 (3H, s), 2.91 (1H, m), 2.70 (1 H, bm), 2.29 (1H, m), 2.21 (1 H, b), 1.28 (3H, d), 1.13 (3H, t): LC/Mass spec, electrospray: +ve m/e: 492 [MH]⁺:

δ Example 99

2-[[((3aS,6S,6aR)-4-[δ-({[(ethylamino)carbonyl] oxy}methyl)-1 ,3-thiazol-2-yl]-6- methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-1-

[(acetyloxy)methyl]ethyl acetate: and 0 Example 100

3-[[((3aS,6S,6aR)-4-[5-({[(ethylamino)carbonyl]oxy}methyl)-1 ,3-thiazoi-2-yl]-6- methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl] (methyl)amino]-2- hydroxypropyl acetate:

26 Example 98 (15.6 mg, 31.7 μmol) was dissolved in dry dichloromethane (1.5 mL) and acetic anhydride (17.97 μL, 190 μmol, 6 equiv) and pyridine (15.4 μL, 190 μmol, 6 equiv) added. The mixture (a solution) was left to stand at room temperature for approx. 6 days before it was washed with 1 M hydrochloric acid (1 mL) and saturated brine (1 mL). The mixture was separated using a hydrophobic

30 frit. The organic phase was evaporated to leave a colourless gum. The gum was purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using ethyl acetate as eluent to give the title compounds Example 99 (9.4 mg, 51%) and Example 100 (3.3 mg 19%): as white solids. Example 99

Η nmr (CDCI₃): δ 7.37 (1 H, s), δ.28 (1 H, m), 5.20 (2H, s), 4.68 (1 H, bm)„4.34 (1H, m), 4.10 (2H, m), 3.82 (1H, t), 3.68 (1 H, dd), 3.45 (3H, m), 3.21 (2H, m), 3.13 (1H, m), 2.95 (3H, 2s), 2.89 (1H, m), 2.26 (1H, m), 2.11 (3H, d), 2.08 (3H, s), 1.27 (3H, δ d), 1.13 (3H, t):

LC/Mass spec, electrospray: +ve m/e: 676 [MH]⁺:

Example 100

^1"H nmr (CDCI₃): δ 7.38 (1H, s), δ.20 (2H, s), 4.66 (1H, bm), 4.10 (4H, m), 3.87 (1H, m), 3.70 (1H, m), 3.54 (1H, m), 3.35 (2H, m), 3.22 (2H, m), 3.16 (1H, m), 3.01 (3H, 0 d), 2.90 (1 H, m), 2.64 (1 H, bm), 2.26 (1H, m), 2.11 (3H, s), 1.28 (3H, d), 1.13 (3H, t):

LC/Mass spec, electrospray: +ve m/e: 534 [MH]⁺:

5 Example 101

(3aS,6S,6aR)-4-(cis-2,3-Dimethyl-cyclopropanecarbonyl)-6-methyl-5-oxo- hexahydro-pyrrolo[3,2-b]pyrrole-1 -sulphonic acid methyl-propyl-amide

Intermediate 16 (27 mg, 0.1 mmol), was dissolved in anhydrous acetonitrile (1 0 mL) and N-methyl-N-propylsulphamoyl chloride (intermediate 37) (17 μL, excess) was added in one portion with stirring, followed by DIPEA (38 μL, 2.2 equiv.). The mixture was stirred at room temperature overnight. The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane and purified by preparative plate chromatography (Whatman 5 PK6F silica gel 60A plate) using 1 :4 ethyl acetate yclohexane as eluent to give the title compound (R_f 0.35) 30 mg (80.7%):

Η nmr (CDCI₃): δ 3.85-3.67 (2H, m), 3.52-3.36 (2H, m), 3.28-2.95 (3H, m), 2.88- 2.79 (4H, m), 2.74-2.62 (1H, m), 2.09-1.91 (1 H, m), 1.70-1.52 (4H, m), 1.28 (3H, d), 1.18 (6H, t), 0.93 (3H, t) 30 Mass spec, thermospray: +ve m/e: 372 [MH]⁺

Example 102 (3aS,6S,6aR)-N-(3-cyanobenzyl)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}- N,6-dimethyl-5-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

Intermediate 46 (20 mg, 60.71 μmol) was dissolved in acetonitrile (1 mL) and δ treated with caesium carbonate (40 mg) and 3-cyanobenzyl bromide (12.6 mg, 1.2 equiv.). The reaction mixture was stirred at room temperature overnight, then filtered and purified by preparative plate chromatography (1 :2 ethyl acetate:cyclohexane) to give the title compound as a colourless solid (23 mg, 85%): 0 Η nmr (CDCI₃) (400 MHz): δ 7.65-7.60 (3H, m), 7.54-7.42 (1 H, m), 4.47-4.36

(2H, m), 3.84-3.77 (2H, m), 3.57-3.53 (1H, m), 3.51-3.44 (1H, m), 3.07-2.99 (1H, m), 2.88-2.70 (5H, m), 2.12-1.99 (1 H, m),1.71-1.δδ (2H, m), 1.27 (3H, d), 1.23- 1.17 (6H. m):

Mass spec, thermospray: +ve m/e: 445 [MH]⁺ 6

Example 103

Similarly prepared to example 102

(3aS,6S,6aR)-N-(2-cyanobenzyl)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}- 0 N,6-dimethyl-5-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

colourless solid (22 mg, 81 %):

Η nmr (CDCI₃) (400 MHz): δ 7.70-7.64 (3H, m), 7.47-7.41 (1 H, m), 4.70-4.67 (2H, m), 3.86-3.77 (2H, m), 3.66-3.48 (2H, m), 3.06-2.98 (1 H, m), 2.87-2.71 (5H, 6 m), 2.11-1.98 (1H, m),1.71-1.56 (2H, m), 1.27 (3H, d), 1.23-1.17 (6H, m):

Mass spec, thermospray: +ve m/e: 445 [MH]⁺

Example 104 30 Similarly prepared to example 102

(3aS,6S,6aR)-4-{r(2R,3S)-2,3-dimethylcyclopropyl1carbonyl}-N-(4- methoxybenzyl)-N,6-dimethyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)- sulphonamide

36 colourless solid (24 mg, 88%): LC-MS (method A):

Peak at 3.64 mins gives m/e 450 [MH⁺]

Example 105

Similarly prepared to example 102

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N-(3- methoxybenzyl)-N,6-dimethyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)- sulphonamide

colourless solid (24 mg, 88%):

LC-MS (method A):

Peak at 3.66 mins gives m/e 460 [MH⁺]

Example 106

Similarly prepared to example 102

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-N-(2- methylbenzyl)-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

colourless solid (23 mg, 87%):

LC-MS (method A):

Peak at 3.88 mins gives m/e 434 [MH⁺]

Example 107

Similarly prepared to example 102

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyciopropyl]carbonyl}-N,6-dimethyl-N-[4- (methylsulphonyl)benzyl]-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)- sulphonamide

colourless solid (20 mg, 66%): LC-MS (method A):

Peak at 3.44 mins gives m/e 498 [MH⁺] Example 108

Similarly prepared to example 102

N-(4-{[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]methyl} phenyi)acetamide

colourless solid (18 mg, 62%): LC-MS (method A): Peak at 3.40 mins gives m/e 477 [MH⁺]

Example 109 Similarly prepared to example 102 ethyl 6-{[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-6- methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl]

(methyl)amino]methyl}-2-furoate

colourless solid (20 mg, 68%): LC-MS (method A): Peak at 3.67 mins gives m/e 482 [MH⁺]

Example 110

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5-oxo- N-(2-oxo-2-phenylethyl)hexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Intermediate 46 (33 mg, 100 μmol) was dissolved in dry THF (1 mL) and treated with potassium t-butoxide (11.2 mg). After stirring at room temperature for 30 mins., phenacyl bromide (20 mg) was added. The reaction mixture was stirred at room temperature overnight, then purified by preparative plate chromatography (1 :2 ethyl acetate:cyclohexane) to give the title compound as a colourless solid (28 mg, 62%): nmr (CDCI₃) (250 MHz): δ 7.96-7.90 (2H, m), 7.68-7.60 (1 H, m), 7.56-7.47 (2H, m), 4.78 (2H, s), 3.84-3.52 (4H, m), 3.06-2.94 (4H, m), 2.87-2.78 (1H, m), 2.75-2.64 (1 H, m), 2.23-1.95 (1 H, m), 1.72-1.50 (2H, m),1.30-1.13 (9H, m) Mass spec, thermospray: +ve m/e: 448 [MH]⁺

Example 111

Similarly prepared to example 110

(3aS,6S,6aR)-N-[2-(4-chlorophenyl)-2-oxoethyl]-4-{[(2R,3S)-2,3- dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5-oxohexahydropyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide

colourless solid (8 mg, 27%): LC-MS (method A): Peak at 3.67 mins gives m/e 482/484 [MH⁺]

Example 112

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-N-(2- methoxyethyl)-6-methyl-5-oxohexahydropyrrolo[3,2-b]pyrroie-1 (2H)- suiphonamide

Intermediate 47 (30 mg, 89.5 μmol) was dissolved in dry DCM (1.0 mL) with DIPEA (17.15 μL, 1.1 equiv) and 2-methoxyethylamine (23.37 μL, 3 equiv) added. The mixture was left to stand at room temperature for 90 hours before it was purified using a SPE 1g silica cartridge eluted with dichloromethane, chloroform, 1 :1 ether: cyclohexane, ether, 1 :1 cyclohexane: ethyl acetate, 1:2 cyclohexane: ethyl acetate, ethyl acetate (χ2) to give the title compound as acolourless gum, 30mg, 89%. LC/MS (A, 5.5min)

Peak at 3.14 min gives m/e 374 [MH]⁺ and 396 [M+Na]⁺.

Example 113 Similarly prepared to example 112 (3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-N-(2-hydroxyethyl)- 6-methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-suIphonamide

Colourless gum 26%

LC/MS (A, δ.δmin)

Peak at 2.86 min gives m/e 360 [MH]⁺ and 382 [M+Na]⁺and 741 [2M+Na]⁺.

Example 114 Similarly prepared to example 112

(3aS,6S,6aR)-N-[2-(benzyloxy)ethyl]-4-{[(2R,3S)-2,3- dimethylcyciopropyl]carbonyl}-6-methyl-5-oxohexahydropyrrolo[3,2-b]pyrrole-

1(2H)-sulphonamide

Colourless gum 79% LC/MS (A, δ.δmin) Peak at 3.63 min gives m/e 450 [MH]⁺.

Example 115

Similarly prepared to example 112

1-[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcycloρropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl]piperidine-4-carboxamide

δ White solid 23%

LC/MS (A, δ.δmin)

Peak at 3.63 min gives m/e 450 [MH]⁺.

0 Example 116

Similarly prepared to example 112

1-[((3aS,6S,6aR)-4-{r(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl]piperidine-3-carboxamide

δ White solid 92% LC/MS (A, 5.5min)

Peak at 3.32 min gives m/e 427 [MH]⁺.

Example 117

Similarly prepared to example 112

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-4-{[4-(1- hydroxyethyl)piperidin-1-yl]sulphonyl}-3-methylhexahydropyrrolo[3,2-b]pyrrol-

2(1 H)-one

White solid 91%

LC/MS (A, δ.δmin)

Peak at 3.14 min gives m/e 428 [MH]⁺ and 450 [M+Na]\

Example 118

Similarly prepared to example 112

(3S,3aR,6aS)-4-{[(3R)-3-(dimethylamino) pyrrolidin-1 -yl]sulphonyl}-1 -{[(2R.3S)- 2,3-dimethylcyclopropyl]carbonyl}-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)- one formate

Pale brown solid 67% LC/MS (A, δ.δmin)

Peak at 2.47 min gives m/e 413 [MH]⁺.

Example 119

(3aS,6S,6aR)-N-allyl-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl -N,6- dimethyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

To intermediate 16 (0.069g, 0.41 mmol) was added a solution of N-allyl-N-methyl sulphamoyl chloride (intermediate 40) (0.1 Og, 0.37mmol) in DCM (10ml) followed by DIPEA (0.1 Oδg, 0.14ml, 0.81 mmol). After 18 hours the reaction mixture was partitioned between ethyl acetate and water. The organic phase was washed with brine and dried over anhydrous MgS0₄. The solvent was removed in vacuo and chromatographed over silica (Merck 9385) using cyclohexane/ethyl acetate

(4:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.077g (56%).

Η NMR (CDCI₃): δ 5.81 (1H, m), 5.29 (1H, m), 5.26 (1H, m), 3.90-3.71 (4H, m),

3.48 (1 H, dd, J=6,10Hz), 3.43 (1 H, m), 3.01 (1 H, pent, J=7Hz), 2.83 (1 H, t,

J=9Hz), 2.85 (3H, s), 2.69 (1 H, m), 2.01 (1H, m), 1.70-1.52 (2H, m), 1.27 (3H, d,

J=7Hz), 1.18 (6H, m).

Mass spec, thermospray: +ve m/e: 370 [MH⁺]

Example 120

(3aS,6S,6aR)-N-(2,3-dihydroxypropyl)-4-{[(2R,3S)-2,3- dimethylcyclopropyl]carbonyl}-N,6-dimethyl-δ-oxohexahydropyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide

To a solution of example 119 (0.069g, 0.19mmol) in THF (2ml) was added a solution of osmium tetroxide in THF (1.34ml of 40mg/ml solution, 0.21 mmol). After 18 hours at room temperature the reaction mixture was separated between ethyl acetate and sodium metabisulphite solution. The organic phase was filtered and dried over anhydrous MgS0₄. The solvent was removed in vacuo and the residue was purified using a Bond-elut cartridge and a cyclohexane/ethyl acetate gradient. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.052g (68%). Η NMR (CDCI₃): δ 3.92 (1H, m), 3.83-3.70 (3H, m), 3.63 (1 H, m), 3.53-3.27 (4H, m), 3.01 (1 H, m), 2.98 (3H, s), 2.83 (1 H, t, J=9Hz), 2.71 (1 H, m), 2.62 (1 H, m),

2.15 (1H, m), 2.01 (1 H, m), 1.70-1.53 (2H, m), 1.27 (3H, d, J=7.5Hz), 1.18 (6H, m). Mass spec, thermospray: +ve m/e: 404 [MH⁺]

Example 121 and Example 122: isomers of (3aS,6S,6aR)-N-(2,3- dihydroxypropyl)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5- oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide Example 120 was separated by preparative HPLC to give two isomers, 0.005g of each.

Example 121 (peak 1 ) δ NMR (CDCI₃): δ 3.91 (1 H, m), 3.82-3.68 (3H, m), 3.63 (1 H, m), 3.53-3.28 (4H, m), 3.06-2.94 (4H, m), 2.82 (1H, t, J=9Hz), 2.71 (1 H, m), 2.62 (1H, m), 2.12 (1H, m), 2.02 (1H, m), 1.70-1.53 (2H, m), 1.27 (3H, m), 1.18 (6H, m).

Example 122 (peak 2) 0 Η NMR (CDCI₃): 53.91 (1 H, m), 3.82-3.68 (3H, m), 3.63 (1 H, m), 3.53-3.27 (4H, m), 3.06-2.94 (4H, m), 2.82 (1 H, t, J=9Hz), 2.71 (1 H, m), 2.02 (1 H, m), 1.70-1.63 (2H, m), 1.27 (3H, m), 1.18 (6H, m).

6 Example 123

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-1- [(acetyloxy)methyl]ethyl acetate

0 To a solution of example 120 (0.020g, O.Oδmmol) in DCM (1 ml) was added pyridine (0.12ml, 0.117g, l .δmmol) and acetic anhydride (0.12ml, 0.130g, 1.3mmol). After 4hours the reaction mixture was separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with saturated sodium bicarbonate solution and brine. The organic phase was dried 5 over anhydrous MgS0₄ and evaporated in vacuo to give the title compound as a colourless gum, 0.022g (90%).

Η NMR (CDCI₃): δ 5.26 (1 H, m), 4.32 (1 H, m), 4.10 (1 H, m), 3.83-3.67 (2H, m), 3.52-3.31 (4H, m), 3.00 (1 H, m), 2.91 +2.94 (3H, s), 2.82 (1 H, t, J=9Hz), 2.70 (1 H, m), 2.11 +2.10+2.08 (6H, s), 2.01 (1 H, m), 1.70-1.50 (2H, m), 1.27 (3H, d,

30 J=7.δHz), 1.18 (6H, m).

Mass spec, thermospray: +ve m/e: 488 [MH⁺]

Example 124 tert-butyl [[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethyl-cyclopropyl]carbonyl}-6- methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)- yl)sulphonyl](methyl)amino]acetate

δ To intermediate 41 (0.246g, 1.01 mmol) was added a solution of intermediate 16 (0.250g, 0.92mmol) in DCM (15ml) and DIPEA (0.262g, 0.35ml, 2.02mmol). After 18 hours the reaction mixture was separated between ethyl acetate and water. The organic phase was washed with brine and dried over anhydrous MgS0₄. The solvent was removed in vacuo and chromatographed over silica 0 (Merck 9385) using cyclohexane/ethyl acetate (4:1 v/v) as eluant. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.311 g (76%).

Η NMR (CDCI₃): δ 3.91 (2H, m), 3.76 (1 H, m), 3.71 (1 H, m), 3.58 (1 H, m), 3.62 (1H, m), 3.01 (3H, s), 2.99 (1 H, m), 2.83 (1 H, t, J=9Hz), 2.67 (1 H, m), 2.02 (1H, 5 m), 1.70-1.54 (2H, m), 1.48 (9H, s), 1.27 (3H, d, J=7Hz), 1.18 (6H, m).

Mass spec, thermospray: +ve m/e: 444 [MH⁺]

Example 125 0 [[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yi)sulphonyl](methyl)amino] acetic acid

To a solution of example 124 (0.084g, 0.19mmol) in DCM (O.δml) was added TFA (O.δml). After 2 hours the solution was poured into saturated sodium

2δ bicarbonate solution. The pH was adjusted to 7 and the mixture was extracted using DCM (x2). The aqueous phase was acidified and extracted using DCM (x2). The later DCM washes were dried over anhydrous MgS0₄and evaporated in vacuo to give the title compound as a colourless gum, 0.020g (27%). Η NMR (CDCI₃): δ 4.08 (2H, m), 3.86-3.68 (2H, m), 3.62-3.47 (2H, m), 3.01 (4H,

30 m), 2.83 (1 H, t, J=9Hz), 2.69 (1 H, m), 2.00 (1 H, m), 1.62 (2H, m), 1.28 (3H, d,

J=7Hz), 1.18 (6H, m). Mass spec, thermospray: +ve m/e: 388 [MH⁺]

35 Alternative synthesis of example 125 benzyl [[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethyl-cyclopropyl]carbonyl}-6-methyl- 5-oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]acetate

A solution of intermediate 45 (4.92g, 10.3mmol) in ethyl acetate (200ml) was hydrogenated over Pd/C (50% water, 1.0g) for 1 hour. The catalyst was removed by filtration and washed with ethyl acetate. The combined filtrate and washings were hydrogenated over Pd/C (50% water, 1.0g) for 2 hours. The catalyst was removed by filtration and washed with ethyl acetate. The combined filtrate and washings were washed with water and brine. The organic phase was dried (anhydrous magnesium sulphate) and evaporated in vacuo to give the title compound as a white solid, 3.40g (85%). nmr (CDCI₃): δ 4.08 (2H, s), 3.85-3.67 (2H, m), 3.61-3.46 (2H, m), 3.03 (3H, s), 3.01 (1 H, m), 2.82 (1 H, t, J=9Hz), 2.69 (1 H, m), 2.00 (1 H, m), 1.62 (2H, m), 1.28 (3H, d, J=7.δHz), 1.18 (6H, m). LC-MS (method A):

Peak at 3.23 mins gives m/e 388 [MH⁺]

Example 126

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- δ oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-[(3- methoxyisothiazol-5-yl)methyl]acetamide

Example 126 (0.020g, 0.0δ2mmol), 3-methoxy-δ-methylaminoisothiazole hydrochloride¹ (0.0094g, 0.0δ2mmol), EDC (0.0125g, 0.065mmol) and HOBT 0 (0.009g, 0.065mmol) were suspended in DCM (5ml). To this was added DIPEA (0.02δg, 0.034ml, 0.19δmmol) and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was separated between ethyl acetate and 2N hydrochloric acid. The organic phase was washed with saturated sodium bicarbonate solution and brine. The organic phase was dried 6 over anhydrous MgS0₄ and evaporated in vacuo. The residue was purified using a Bond-elut cartridge and a cyclohexane/ethyl acetate gradient. The required fractions were combined and evaporated in vacuo to give the title compound as a white solid, 0.011 g (41%).

Η NMR (CDCI₃): δ 6.90 (1H, m), 6.49 (1H, s), 4.65 (2H, m), 3.98 (3H, s), 3.91

(2H, m), 3.82-3.72 (2H, m), 3.55-3.43 (2H, m), 3.00 (1 H, m), 2.98 (3H, s), 2.82

(1 H, t, J=9Hz), 2.73 (1 H, m), 2.04 (1 H, m), 1.70-1.54 (2H, m), 1.27 (3H, d,

J=7.δHz), 1.18 (6H, m).

Mass spec, thermospray: +ve m/e: 514 [MH⁺]

¹Ref. 1 Lykkeberg.J.; Krogsgaard-Larsen.P.; Acta Chem.Scand.Ser.B; 30; 1976; 781-785

Example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-[(1S)-1- (hydroxymethyl)-3-(methylthio)propyl]acetamide

Example 125 (20mg, 0.052mmol), EDC (11 mg, 0.067mmol), HOBT (10.5mg, 0.077mmol), DIPEA (19.8μl, 0.11 mmol) were dissolved in acetonitrile (1ml). (S)- methioninol (7.7mg, 0.0δ7mmol) was added and the mixture left for 18 hours at room temperature. The acetonitrile was removed with a stream of nitrogen and the residue dissolved in dichloromethane (1 ml), washed with 0.1 M aqueous HCI (0.5ml), separated then washed with saturated aqueous sodium hydrogen carbonate (1 ml) and passed through a hydrophobic frit. The collect organic phase was purified using preparative plate chromatography (Whatman PK6F silica 60A plate) using neat ethyl acetate as the eluent to give the title compound, white solid, 13.2mg (51%)

LC-MS (A)

Peak at time 3.15minutes gave m/e 505 (M+H)⁺

Example 128

Similarly prepared to example 127 2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-[(1S)-2- hydroxy-1 -phenylethyljacetamide

White solid, 13.8mg (49%) LC-MS (A) Peak at time 3.27 minutes gave m/e 607 (M+H)⁺

Example 129

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethyicyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-[(1R)-2- hydroxy-1 -phenylethyljacetamide

White solid, 12.9mg (49%)

LC-MS (A)

Peak at time 3.25 minutes gave m/e 507 (M+H)⁺

Example 130

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)aminoj-N-[(1 R)-1- benzyl-2-hydroxyethyl]acetamide

White solid, 11.6mg (43%)

LC-MS (A)

Peak at time 3.24 minutes gave m/e 521 (M+H)⁺

Example 131

Similarly prepared to example 127

2-r[((3aS,6S,6aR)-4-{ 2R,3S)-2,3-dimethylcyclopropyljcarbonyl}-6-methyl-5- oxohexahydropyrrolor3,2-bjpyrrol-1(2H)-yl)sulphonylj(methyl)aminoj-N-[(1S)-1- benzyl-2-hydroxyethyljacetamide White solid, 14.6mg (54%)

LC-MS (A)

Peak at time 3.32 minutes gave m/e 521 (M+H)⁺

Example 132

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyljcarbonyI}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1(2H)-yl)sulphonylj(methyl)amino]-N-[1-(4- chlorobenzyl)-2-hydroxyethyljacetamide

White solid, 10.3mg (36%) LC-MS (A) Peak at time 3.49 minutes gave m/e 555 (M+H)⁺ and m/e 557 (M+H)⁺

Example 133

Similarly prepared to example 127 2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1 (2H)-yl)sulphonylj(methyl)aminoj-N-[(1S)-2- hydroxy-1-(4-nitrobenzyl)ethyljacetamide

(S)-2-amino-3-(4-nitrophenyl) propanol prepared according to the method of Cazin et al. Journal of the Chemical Society, Perkin Transactions 1 1989, 867-872

White solid, 12..4mg (42%)

LC-MS (A)

Peak at time 3.36 minutes gave m/e 566 (M+H)⁺

Example 134

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyljcarbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)aminoj-N-[(1R)-2- hydroxy-1 -(4-nitrobenzyl)ethyljacetamide (R)-2-amino-3-(4-nitrophenyl) propanol prepared according to the method of Donnow and Winter, Chem. Ber. 1951 , 51. 307-311

White solid, 11.8mg (40%)

LC-MS (A)

Peak at time 3.36 minutes gave m/e 566 (M+H)⁺

Example 135 2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyljcarbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1 (2H)-yl)sulphonylj(methyl)aminoj-N-[(1 R)-1- (hydroxymethyl)-2-methylpropyl]acetamide

Example 125 (20mg, 0.052mmol), EDC (11 mg, 0.057mmol), HOBT (10.δmg, δ 0.077mmol), DIPEA (19.8μl, 0.11 mmol) were dissolved in acetonitrile (1ml). (S)- 2-amino-3-phenyl-1 -propanol (8.6mg, 0.0δ7mmol) was added and the mixture left for 18 hours at room temperature. Further additions of EDC (11mg, 0.0δ7mmol), DIPEA 10μl, O.Oδδmmol) and (S)-2-amino-3-phenyl-1 -propanol (8.6mg, 0.057mmol) were made and the mixture left overnight. The acetonitrile 0 was removed with a stream of nitrogen and the residue dissolved in dichloromethane (1 ml), washed with 0.1 M aqueous HCI (O.δmi), separated then washed with saturated aqueous sodium hydrogen carbonate (1 ml) and passed through a hydrophobic frit. The collect organic phase was purified using preparative plate chromatography (Whatman PK6F silica 60A plate) using neat δ ethyl acetate as the eluent to give the title compound, white solid, 8.6mg (36%) LC-MS (A) Peak at time 3.1 δ minutes gave m/e 473 (M+H)⁺

0 Example 136

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1(2H)-yl)sulphonylj(methyl)amino]-N-(3- nitrobenzyl)acetamide 6 colourless solid (16 mg, 61%): nmr (CDCI₃) (400 MHz): δ 8.16-8.13 (2H, m), 7.65-7.62 (1 H, m), 7.56-7.50

(1H, m), 6.99 (1 H, broad t), 4.59 (2H, d, J 7Hz), 4.01-3.88 (2H, m), 3.82-3.74

(2H, m), 3.55-3.45 (2H, m), 3.03-2.96 (4H, m), 2.82 (1 H, t, J 9Hz)), 2.76-2.68

(1 H, m), 2.09-1.98 (1 H, m), 1.70-1.55 (2H, m), 1.26 (3H, d, J 7Hz), 1.21-1.16

(6H, m):

LC-MS (method A):

Peak at 3.35 mins gives m/e 622 [MH⁺]

Example 137

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyljcarbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1 (2H)-yl)sulphonylj(methyl)amino]-N-(3,5- difluorobenzyl)acetamide

colourless solid (12 mg, 47%): LC-MS (method A):

Peak at 3.44 mins gives m/e 513 [MH⁺]

Example 138

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-bjpyrrol-1(2H)-yl)sulphonylj(methyl)amino]-N-[3,5- bis(trifluoromethyl)benzyl]acetamide

colourless solid (27 mg, 88%): LC-MS (method A): Peak at 3.74 mins gives m/e 613 [MH⁺]

Example 139

Similarly prepared to example 127 2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-δ- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl](methyl)amino]-N-(4- bromobenzyl)acetamide

colourless solid (2δ mg, 90%):

LC-MS (method A):

Peak at 3.45 mins gives m/e 565/657 [MH⁺]

Example 140

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(4- iodobenzyl)acetamide

colourless solid (30 mg, 99%):

LC-MS (method A):

Peak at 3.57 mins gives m/e 603 [MH⁺]

Example 141

Similarly prepared to example 127

2-[[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulphonyl](methyl)amino]-N-(2,4- dimethoxy-5-nitrobenzyl)acetamide

colourless solid (23 mg, 79%):

Η nmr (CDCI₃) (400 MHz): δ 7.95 (1 H, s), 6.9δ (1 H, broad t), 6.50 (1 H, s), 4.42 (2H, d, J 7Hz), 3.99 (3H, s), 3.96 (3H, s), 3.94-3.83 (2H, m), 3.80-3.72 (2H, m), 3.54-3.44 (2H, m), 3.03-2.96 (4H, m), 2.82 (1 H, t, J 9Hz)), 2.74-2.66 (1 H, m),

2.08-1.97 (1 H, m), 1.70-1.56 (2H, m), 1.26 (3H, d, J 7Hz), 1.21-1.16 (6H, m): LC-MS (method A): Peak at 3.57 mins gives m/e 582 [MH⁺] Example 142 rel-(3aS,6S,6aR)-4-Cycloρropanecarbonyl-6-methyl-5-oxo-hexahydro- pyrrolo[3,2-b1pyrrole-1 -sulphonic acid isopropyl-methyl-amide

The racemic intermediate 13 (49 mg, 0.2 mmol) was stirred in anhydrous acetonitrile (1 mL) and N-methyl-N-isopropylsulphamoyl chloride (intermediate 38) (51 mg, 0.3 mmol.) was added in one portion, followed by DIPEA (62 mg, 0.4 mmol, 70 μL). The mixture was stirred at room temperature overnight. The solvent was removed using a stream of nitrogen, and the residual gum was taken up in dichloromethane and purified by preparative plate chromatography (Whatman PK6F silica gel 60A plate) using 1 :2 ethyl acetate yclohexane as eluent to give the title compound (R_f 0.6) as a colourless solid 32 mg (46.5%): nmr (CDCI₃): δ 4.14 (1 H, septet), 3.84-3.65 (2H, m), 3.53-3.34 (2H, m), 3.11- 2.88 (2H, m), 2.76 (3H, s), 2.72-2.58 (1 H, m), 2.12-1.94 (1 H, m), 1.29 (3H, d), 1.20 (6H, d), 1.20-0.96 (4H, m)

Mass spec, thermospray: +ve m/e: 344 [MH]⁺

Example 143 Similarly prepared to example 142 rel-(3aS,6S,6aR)-4-Cyclopropanecarbonyl-6-methyl-5-oxo-hexahydro- pyrrolo[3,2-b]pyrrole-1 -sulphonic acid propyl-methyl-amide colourless solid (56%) nmr (CDCI₃): δ 3.84-3.68 (2H, m), 3.54-3.36 (2H, m), 3.28-2.85 (7H, m), 2.73- δ 2.67 (1 H, m), 2.13-1.94 (1 H, m), 1.71-1.56 (2H, m), 1.28 (3H, d), 1.20-0.89 (7H, m).

Mass spec, thermospray: +ve m/e: 344 [MH]⁺

0 Example 144

(3aS,6S,6aR)-4-(cis-2,3-Dimethyl-cyclopropanecarbonyl)-6-methyl-5-oxo- hexahydro-pyrrolo[3,2-b]pyrrole-1 -sulphonic acid isopropyl-methyl-amide 136

Intermediate 48 (39 mg, 0.143 mmol) in anhydrous THF (1 mL) was stirred under dry nitrogen at -10°C and was treated with a solution of lithium hexamethyldisilazide (143 μL of 1.0 M solution in THF). After 20 minutes at -10°C cis- cis- 2,3-dimethylcyciopropylcarboxylic acid anhydride (prepared from the corresponding acid¹ by the method of Nangia and Chandrasekaran² )

(30 mg, 0.143 mmol) in anhydrous THF (1 mL) was added. The mixture was stirred at -10 to 0°C for 2 hours then shaken with ethyl acetate and saturated aqueous sodium hydrogen carbonate (20 mL each). The organic phase was separated, washed with water, dried over anhydrous magnesium sulphate, filtered and evaporated in vacuo to give a yellow solid; this was purified by preparative piate chromatography (Whatman PK6F silica gel 60A plate) using 1:2 ethyl acetate:cyclohexane as eluent to give the title compound as a colourless solid (Rf 0.65) 44 mg (83%): Η nmr (CDCI₃): δ 4.14 (1 H, septet), 3.84-3.65 (2H, m), 3.51-3.32 (2H, m), 3.07- 2.95 (1 H, m), 2.84 (1 H, dd), 2.76 (3H, s), 2.74-2.62 (1H, m), 2.09-1.91 (1H, m),

1.62-1.30 (2H, m), 1.29 (3H, d), 1.26-1.16 (12H, m) Mass spec, thermospray: +ve m/e: 372 [MH]⁺

Ref. 1. L.B. Rodewald and M.C. Lewis; Tet. Lett. 27 (1971 ) 5273 Ref. 2. A. Nangia and S. Chandrasekaran; J. Chem. Res. (1984), 100

Example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4-

(piperidin-1-ylsulphonyl)hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

To a solution of intermediate 47 (30mg, 0.09mmol) in DCM (1 ml) was added DIPEA (13mg, 17.5μL. O.l mmol) and piperidine (7.7mg, 8.9μL, 0.09mmol). After 48 hours the solution was applied to a preparative t.l.c. plate and eluted with cyclohexane/ethyl acetate (2:1 v/v). The required band was removed and eluted with ethyl acetate. The solvent was removed in vacuo to give the title compound as a white solid (27mg, 78%). LC-MS (method A): Peak at 3.50 mins gives m/e 384 [MH⁺] Example 146

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4- (morpholin-4-ylsulphonyl)hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

White solid (26mg, 75%):

Η nmr (CDCI₃): δ 3.82-3.72 (6H, m), 3.53-3.42 (2H, m), 3.29 (4H, m), 3.02 (1 H, m), 2.83 (1 H, t, J=9Hz), 2.71 (1 H, m), 2.02 (1 H, m), 1.70-1.52 (2H, m), 1.27 (3H, d, J=7.5Hz), 1.18 (6H, m) LC-MS (method A): Peak at 3.14 mins gives m/e 386 [MH⁺]

Example 147

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethyicyclopropyl]carbonyl}-4-[(4- hydroxypiperidin-1-yl)sulphonyl]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)- one

White solid (20mg, 84%):

Η nmr (CDCI₃): δ 3.91 (1 H, m), 3.82-3.71 (2H, m), 3.63-3.54 (2H, m), 3.50-3.40 (2H, m), 3.21-3.10 (2H, m), 3.00 (1 H, m), 2.83 (1 H, t, J=9Hz), 2.68 (1 H, m), 2.07-1.90 (3H, m), 1.70-1.48 (4H, m), 1.27 (3H, d, J=7.5Hz), 1.18 (6H, m). LC-MS (method A):

Peak at 2.96 mins gives m/e 400 [MH⁺]

Example 148 Similarly prepared to example 145

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-5-oxo- N,N-dipropylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

White solid (5.5mg, 23%): LC-MS (method A): Peak at 3.86 mins gives m/e 400 [MH⁺]

Example 149 Similarly prepared to example 145

(3aS,6S,6aR)-N-benzyl-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-N,6- dimethyl-5-oxohexahydropyrrolo[3,2-b]pyrroie-1(2H)-sulphonamide

White solid (22.2mg, 88%): LC-MS (method A):

Peak at 3.62 mins gives m/e 420 [MH⁺]

Example 150 Similarly prepared to example 145

(3aS,6S,6aR)-N-benzyl-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-N-(2- hydroxyethyl)-6-methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)- sulphonamide

White solid (9mg, 45%):

LC-MS (method A): Peak at 3.60 mins gives m/e 460 [MH⁺]

Example 151

Similarly prepared to example 145

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N-(2-methoxyethyl)-

N,6-dimethyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulphonamide

Colourless gum (14mg, 80%): LC-MS (method A): Peak at 3.32 mins gives m/e 388 [MH⁺]

Example 152 Similarly prepared to example 145

(3aS,6S,6aR)-N-{2-[(dimethylamino)sulphonyl]ethyl}-4-{[(2R,3S)-2,3- dimethylcyclopropyl]carbonyl}-6-methyl-5-oxohexahydropyrrolo[3,2-b]pyrrole- 1(2H)-sulphonamide

White solid (3.7mg, 14%):

LC-MS (method A):

Peak at 3.10 mins gives m/e 451 [MH⁺]

Example 153

Similarly prepared to example 145 tert-butyl 3-{[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-6- methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl]amino}-2- methylpropanoate

White solid (15.1 mg, 56%): LC-MS (method A):

Peak at 3.58 mins gives m/e 458 [MH⁺]

Example 154

Similarly prepared to example 146

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyi]carbonyl}-3-methyl-4- (piperazin-1 -ylsulphonyl)hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

white solid (23mg) LC/MS (method A)

Peak at 2.48 mins gives m/e 385 [MH⁺]

Example 155

Similarly prepared to example 145 (3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethyicyclopropyl]carbonyl}-4-[(3- hydroxypiperidin-1-yl)sulphonyi]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)- one

colourless gum (26mg)

LC/MS (method A)

Peak at 3.13 mins gives m/e 400 [MH⁺]

Example 156

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4-[(4- methyl-1 ,4-diazepan-1 -yl)sulphonyl]hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

white solid (23mg) LC/MS (method A) Peak at 2.51 mins gives m/e 413 [MH⁺]

Example 157

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4-[(4- pyridin-2-ylpiperazin-1-yl)sulphonyl]hexahydropyrrolo[3,2-b]pyrrol-2(1H)-one

white solid (2δmg) LC/MS (method A) Peak at 3.25 mins gives m/e 462 [MH⁺]

Example 158

Similarly prepared to example 145

N-{1-[((3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl] carbonyl}-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulphonyl] pyrrolidin-3-yl}acetamide

colourless gum (27mg) LC/MS (method A)

Peak at 2.99 mins gives m/e 427 [MH⁺]

Example 159

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-4-[(3- hydroxypyrrolidin-1-yl)sulphonyl]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)- one

colourless gum (23mg)

LC/MS (method A)

Peak at 3.03 mins gives m/e 386 [MH⁺]

Example 160

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethyicyclopropyl]carbonyl}-3-methyl-4-[(4- phenylpiperazin-1-yl)sulphonyl]hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

colourless gum (27mg)

LC/MS (method A)

Peak at 3.8 mins gives m/e 461 [MH⁺]

Example 161

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyi}-4-{[3- (hydroxymethyl)piperidin-1-yl]sulphonyl}-3-methylhexahydropyrrolo[3,2-b]pyrrol- 2(1 H)-one

colourless gum (26mg)

LC/MS (method A)

Peak at 3.21 mins gives m/e 414 [MH⁺] Example 162

Similarly prepared to example 145

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4- (pyrrolidin-1-ylsulphonyl)hexahydropyrrolo[3,2-b]pyrrol-2(1H)-one

colourless gum (26mg) LC/MS (method A)

Peak at 3.43 mins gives m/e 370 [MH⁺]

Example 163

Similarly prepared to example 145

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-6-methyl-N-(2- methylbenzyl)-5-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-suIphonamide

white solid (8.2mg) LC/MS (method A)

Peak at 0.00 mins gives m/e 000 [MH⁺]

Example 164

Similarly prepared to example 145

(3aS,6S,6aR)-N-(3-cyanobenzyl)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}- 6-methyl-5-oxohexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulphonamide

colourless gum (17mg) LC/MS (method A)

Peak at 0.00 mins gives m/e 000 [MH⁺]

Example 165

Similarly prepared to example 145 (3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N-[2-fluoro-5-

(trifluoromethyl)benzyl]-6-methyl-δ-oxohexahydropyrrolo[3,2-b]pyrrole-1(2H)- sulphonamide

white solid (7mg)

LC/MS (method A)

Peak at 0.00 mins gives m/e 000 [MH⁺]

Example 166

(3aS,6S,6aR)-N-{4-[(cyclopropylamino)methyl]benzyl}-4-{[(2R,3S)-2,3- dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5-oxohexahydro pyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide

δ Intermediate 60 (20mg, 0.039mmol) was dissolved in DMF (0.6ml) and treated with potassium carbonate (11mg, O.Oδmmol). Cyclopropylamine (3.4mg, 0.059mmol, 4.1 ul) was added and the reaction allowed to stir at room temperature for 3 days. The DMF was removed in vacuo and the residue dissolved in DCM (3ml) and washed with water (2ml). The organic phase was 0 separated and concentrated and the residue purified by preparative HPLC using

FCAL2 to give the title compound as a colourless gum (2.2mg) LC/MS (method A) Peak at 2.7 mins gives m/e 489 [MH⁺]

δ

Example 167

Similarly prepared to example 166

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-N-[4- (morpholin-4-ylmethyl)benzyl]-δ-oxohexahydropyrrolo[3,2-b] pyrrole-1(2H)- 0 sulphonamide

white solid (14.1 mg) LC/MS (method A)

Peak at 2.78 mins gives m/e 619 [MH⁺] 36 Example 168

Similarly prepared to example 166

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-N-{4- δ [(4-methylpiperazin-1 -yl)methyl]benzyl}-δ-oxohexahydropyrrolo [3,2-b]pyrrole- 1 (2H)-sulphonamide

white solid (17mg) LC/MS (method A) 0 Peak at 2.78 mins gives m/e 632 [MH⁺]

Example 169

Similarly prepared to example 166 6 (3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N-(4-{[ethyl(2- hydroxyethyl)amino]methyl}benzyi)-N,6-dimethyl-5-oxohexahydro pyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide

colourless gum (17.8mg) 0 LC/MS (method A)

Peak at 2.75 mins gives m/e 521 [MH⁺]

Example 170 δ Similarly prepared to example 166

(3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N-(4-

{[dimethylamino]methyl}benzyl)-N,6-dimethyl-5-oxohexahydro pyrrolo[3,2- b]pyrrole-1 (2H)-sulphonamide white solid (9.3mg) 30 LC/MS (method A)

Peak at 2.64 mins gives m/e 477 [MH⁺]

Example 171 35 Similarly prepared to example 166 (3aS,6S,6aR)-4-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-N,6-dimethyl-5-oxo- N-[4-(pyrrolidin-1 -ylmethyl)benzyl]hexahydropyrrolo[3,2-b] pyrrole-1 (2H)- sulphonamide

white solid (13.4mg)

LC/MS (method A)

Peak at 2.79 mins gives m/e 503 [MH⁺]

Example 172 (3S,3aR,6aS)-1 -(1 ,3-benzothiazol-2-yl)-3-methyl-4-[(1 - methylbutyl)sulfonyl]hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

Similarly prepared to example 1 , using 2-chlorosulphonylpentane (intermediate 63):

LC/MS (method A)

Peak at 3.56 mins gives m/e 408 [MH⁺]

Example 173 (3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-3-methyl-4-(pent-4- enylsulfonyl)hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

Intermediate 21 (3S,3aR,6aS)-1-Benzothiazol-2-yl-3-methyl-hexahydro-pyrrolo [3,2-b]pyrrol-2-one trifluoroacetate (118 mg) in dry DCM was treated with triethylamine (209 μL, 5 equiv.) followed by pent-4-enyl-1 -sulphonyl chloride¹ (103 mg, 2 equiv.). After four hours at room temperature, water (δ mL) was added and the organic phase was separated using a phase separation cartridge. The filtrate was evaporated in vacuo, and the residue was purified by Bond-Elut cartridge (δ g cartridge) to give the title compound as a colourless solid (81 mg, 66%) Rf 0.3 on silica using DCM:diethyl ether 48:2 as eluent: Mass spectrum: m/e 406 [MH]⁺ Ref1. King, J.F.; Harding, D.R.K; J.Amer.Chem.Soc. 98; 11 ; (1976) 3312-3316 Example 174: 4-[((3aS.6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulfonyl]butanal

Example 173 (75 mg, 0.185 mmol) was dissolved in DCM (15 mL) andthe solution was cooled to -70°C. Ozone was bubbled through the solution until a blue colour persisted. Oxygen was then bubbled through the solution for 1 hour, followed by nitrogen for 30 minutes. Triphenylphosphine (73 mg) was then added, and the mixture was allowed to warm to room temperature. After testing for disappearance of peroxides (Merckquant test paper) the mixture was concentrated in vacuo and the residue was purified on a 5 g Bond-Elut cartridge to give the title compound as a colourless foam (62%), Rf 0.6 on silica (DCM.diethyl ether 1 :1 ): Mass spectrum: m/e 408 [MHf

Example 175: (3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-4-[(3-{4- morpholino}propyl)sulphonyl]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1H)-one

To a solution of morpholine (33 μL, 374 μmol, 4 equiv.) in dry acetonitrile (1 mL), stirred at room temperature under nitrogen was added potassium carbonate (14 mg, 103 μmol, 1.1 equiv.) and a solution of intermediate 59 (47.2 mg, 93.4 μmol) in dry acetonitrile (1 mL). The reaction mixture was stirred under nitrogen for 8 hours at room temperature and then filtered. The filtrate was diluted with ethyl acetate (20 mL) and washed with 2M HCI solution (20 mL), water (20 mL), and saturated brine (20 mL). After drying over magnesium sulphate the organic phase was evaporated in vacuo to give a yellow residue (5 mg). The water phase was found to contain some of the title compound, and was freeze-dried to give a yellow residue which was taken up in DCM and evaporated in vacuo to give the title compound as a yellow foam (17 mg, 39%). The acidic aqueous extract was basified with solid sodium hydrogen carbonate and extracted with ethyl acetate (2 x 20 mL). The solvent was evaporated in vacuo and the residue purified by Bond-Elut silica cartridge (1 g), eluting with DCM, chloroform and finally with ethyl acetate. The ethyl acetate fractions were evaporated in vacuo to afford the title compound as a yellow foam (22.3 mg, 43%). (total yield 89%). LC/MS (method A) Peak at 2.47 mins gives m/e 466 [MH⁺] Example 176: (3S,3aR,6aS)-1-(1 ,3-benzothiazol-2-yl)-4-[(3- aminopropyl)sulphonyl]-3-methylhexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

Intermediate 64 (51.1 mg, 121.5 μmol) in IPA (3 mL) and DCM (2 mL) was added to 10% palladium on carbon (38 mg, Degussa type catalyst). 1 M HCI in diethyl ether (0.5 mL) was added, and the mixture was stirred at room temperature under an atmosphere of hydrogen for 3 hours. The catalyst was removed by filtration and the filter papers and catalyst were stirred with IPA and methanol for 15 minutes. The filtrate and washings were evaporated in vacuo to give the title compound as a white solid (27.4 mg, δ2%): LC/MS (method A) Peak at 2.43 mins gives m/e 396 [MH⁺]

Example 177: [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino]sulfonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-benzothiazol-6-yl]methyl methylcarbamate

Example 94 (57.5 mg, 131.1 μmol) in dry THF (4 mL) was treated with triethylamine (18.3 μL, 1 equiv.) and methyl isocyanate (77 μL), 10 equiv.). The mixture was stirred at room temperature under nitrogen for 48 hours, then evaporated to dryness in vacuo. Purification by preparative tic on silica (20:40:1 ethyl acetate:toluene:acetic acid eluent, running plate three times, drying well between runs) gave the title compound as a white solid (48.9 mg, 75.3%): LC/MS (method A) Peak at 3.37 mins gives m/e 496 [MH⁺]

Example 178 [2-((3S,3aR,6aS)-3-methyl-4-{fmethyl(propyl)amino]sulfonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-benzothiazol-6-yl]methyl acetate

Example 94 (41.4 mg, 94.4 μmol) in dry acetonitrile (3 mL) was treated with pyridine (8.4 μL, 1.1 equiv.) and acetic anhydride (9.8 μL), 1.1 equiv.), followed by DMAP (3 mg). The mixture was heated at reflux under nitrogen for δ hours, then partitioned between water (15 mL) and DCM (25 mL). The organic phase was washed with 2M HCI solution (15 mL) and water (1δ mL), dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was crystallised from 3:7 ethyl acetate:cyclohexane to give the title compound as a white solid (27.8 mg, 61.3%): LC/MS (method A) Peak at 3.41 mins gives m/e 481 [MH⁺]

Example 179: N-{[2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino1sulfonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)-1 ,3-benzothiazol-6- yl]methyl}acetamide:

δ To a stirred solution of intermediate 65 (23.3 mg, 38.7 μmol) in THF at room temperature was added a 1.0 M solution of TBAF in THF (42.6 μL, 1.1 equiv.). The solution was stirred for 10 minutes, then treated with acetic anhydride (7.3 μL, 2 equiv.). Further aliquots of 1.0 M TBAF solution in THF (42.6 μL each) were added after 2.5 hours and 24 hours. After a further 6 hours, the mixture 0 was partitioned between saturated brine (10 mL) and ethyl acetate (10 mL). The organic phase was dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was purified by preparative tic on silica (7:3 ethyl acetate:cyclohexane eluent) to give the title compound as a white solid (7.5 mg, 40.4%): δ LC/MS (method A)

Peak at 4.42 mins gives m/e 480 [MH⁺]

Example 180: (3aS,6S,6aR)-4-(4-chloro-1 ,3-benzothiazol-2-yl)-N,6-dimethyl-5- 0 oxo-N-propylhexahydropyrrolof3,2-b]pyrrole-1(2H)-sulfonamide:

Prepared similarly to example 8 by coupling intermediate 26 and 2-bromo-7- chlorobenzothiazole¹ to give the title compound as a white solid (54%): LC/MS (method A) 35 Peak at 3.68 mins gives m/e 443, 446 [MH⁺] Ref. 1 R C Elderfield and F W Short; J Org. Chem. 1953, 18, 1092-1103

Example 181 (3aS,6S,6aR)-4-(4-chloro-1 ,3-benzothiazol-2-yl)-N,6-dimethyl-δ- δ oxo-N-propylhexahydropyrrolo[3,2-b]pyrrole-1 (2H)-sulfonamide:

Example 68 (30 mg) in DCM (1 mL) was treated with triethylamine (7.8 mg, 11 μL, 1 equiv.) then with allyl isocyanate. The mixture was allowed to stand at room temperature for 3 days, then purified by preparative tic on silica (1 :1 ethyl 0 acetate:cyclohexane eluent) to give the title compound as a white solid (26 mg, 71%):

LC-MS (method B) Peak at 4.41 mins gives m/e 472 [MH⁺]

5

Example 182 [2-((3S,3aR,6aS)-3-methyl-4-{[methyl(propyl)amino1sulfonyl}-2- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)-1 ,3-thiazol-δ-yl]methyl propylcarbamate:

0 Prepared similarly to example 181 white solid (71%) LC-MS (method B) Peak at 4.49 mins gives m/e 472 [MH⁺]

2δ

Example 183 2-[[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)sulfonyl](methyl)amino]-N-(2- morpholin-4-ylethyl)acetamide formate (1 :1 )

30 Prepared similarly to example 25 purified on reversed-phase HPLC using fcal2: white solid (51 %):

LC-MS (method A)

Peak at 2.49 mins gives m/e 537 [MH⁺] 35 Example 184 2-[({2-r[((3aS,6S,6aR)-4-(1 ,3-benzothiazol-2-yl)-6-methyl-5- oxohexahydropyrrolo[3,2-b]pyrrol-1 (2H)-yl)sulfonyl](methyl)amino1 acetyl}amino)methyl]-4-methyl-1 ,3-thiazole-δ-carboxamide:

Prepared similarly to example 25 white solid (23%):

LC-MS (method B)

Peak at 4.37 mins gives m/e 537 [MH⁺]

Example 185:

(3S,3aR,6aS)-1-{[(2R,3S)-2,3-dimethylcyclopropyl]carbonyl}-3-methyl-4-[(2- phenylpiperidin-1-yl)sulfonyl]hexahydropyrrolo[3,2-b]pyrrol-2(1 H)-one

Prepared similarly to example 145

White solid (67%) LC-MS (method A) Peak at 3.73 mins gives m/e 456 [MH⁺]

Example 186:

(3aS,6S,6aR)-4-trimethylacetyl-N,6-dimethyl-5-oxo-N- propylhexahydropyrrolo[3,2-b]pyrrole-1(2H)-sulfonamide

Intermediate 26 (50.6 mg, 183 μmol) in dry THF (2.5 mL) under nitrogen at -

70°C was treated with lithium hexamethyldisilazide (0.24 mL of 1.0 M solution in THF, 1.3 equiv.) and the solution was stirred at -70°C for 5 minutes, then at 0°C for 5 minutes and cooled again to -70°C Trimethylacetyl chloride (68 μL, 3 equiv.) was added and the mixture was stirred at -70°C for 2 hours, then quenched by addition of saturated aqueous ammonium chloride (2.5 mL). The mixture was allowed to warm to room temperature, diluted with saturated aqueous ammonium chloride (10 mL) and extracted with ethyl acetate (10 mL). The organic phase was washed with water (6 mL), saturated brine (10 mL), dried over anhydrous magnesium sulphate, filtered and evaporated under reduced pressure. The crude product was purified using a Bond-elut silica cartridge (2g) eluting successively with cyclohexane, 2:1 cyclohexane:diethyl ether, 1 :1 cyclohexane:diethyl ether, 1:2 cyclohexane:diethyl ether and diethyl ether. The 2:1 cyclohexane:diethyl ether and 1 :1 cyclohexane:diethyl ether fractions were combined and evaporated in vacuo to give the title compound as a white solid (56.6 mg, 89.7%):

LC-MS (method A)

Peak at 3.48 mins gives m/e 360 [MH⁺]

Certain of the compounds according to formula (I) which are exemplified above have been tested in HSV-1 , HSV-2 and hCMV serine protease enzyme inhibition assays and in anti-viral Elisa or plaque reduction assays against HSV-2, HSV-1 , VZV and hCMV, as well in as a Vero cell cytotoxicity assay. The data is shown in Table 1 below.

General Methods

Antiviral assays.

1). Plaque reduction assays.

Human cytomegalovirus (HCMV)

Monolayers are formed by seeding 24-well tissue culture plates with 10⁵_MRC-5 human fibroblasts per well suspended in Dulbecco's modification of Eagle's medium (DMEM) supplemented with 10% foetal calf serum (FCS), 1% non- essential amino acid solution and 2mM L-glutamine. Following incubation at 37°C overnight in a δ% C0₂ atmosphere and subsequent removal of the growth medium, the cells are infected with 0.2ml of HCMV (strain AD169) suspension containing approximately 100 plaque-forming units. They are then maintained at 37°C for 1 hour prior to overlaying with DMEM containing 4% FCS and 1% carboxymethyl-cellulose. After 53 hours further incubation at 37°C in 5% C0₂, the original overiay is removed by aspiration and replaced by a similar overiay containing serial doubling dilutions of the test drug in the range 100-1.δ6μM, freshly prepared from a 40mM stock dissolved in dimethyl sulphoxide. Two 161

further replacements with overlay containing freshly prepared dilutions of drug are carried out at 72 & 77 hours post infection. On day 6 the cell sheets are fixed with 10% formol saline and stained with 0.3% methylene blue. Plaques are counted microscopically, and the mean count for duplicate wells at each drug concentration is calculated as a percentage of the drug-free virus control wells. The 60% inhibitory concentration of the test compound is calculated by regression analysis of the curve of percentage plaque reduction against drug concentration.

Herpes simplex virus (HSV).

The assay for HSV is broadly similar to that for HCMV with the following modifications;

(a) Vero cells grown in DMEM supplemented with 2mM L-glutamine and δ% δ FCS replace the MRC-δs.

(b) The duration of the assay is reduced to 48 h.p.i. for both HSV 1 (strain SC16) and HSV 2 (strain 186), with a delayed addition of drug dilutions in DMEM + 2% FCS at 5.5 h.p.i.

(c) The cell monolayers are fixed using a 5% aqueous solution of glutaraldehyde 0 and stained with carbol fuchsin.

Varicella Zoster Virus (VZV)

The assay for VZV is also broadly similar to that for HCMV with the following δ modifications;

(a) MRCδ cells are infected with 0.2ml of VZV strain G31 at 37°C for 90min.

(b) The overlay does not contain carboxymethyl cellulose. Drug dilutions are added in the initial overlay at 90 minutes post-infection.

(c) The cell monolayers are fixed as for HCMV at 96 hours post infection. 0

2). Enzyme Linked ImmunoSorbant Assays (ELISA).

HCMV 35 Tissue culture:

Compounds are formulated to 40mM in DMSO, then further diluted to 4 times the highest required final concentration in assay medium (bicarbonate-buffered Dulbecco's modification of Eagle's medium (DMEM) supplemented with δ% v/v foetal calf serum, 2mM w/v glutamine and antibiotics). 100μl of four test compounds are transferred to 3 wells each in the first row of a 96-well, plastic, tissue-culture plate, and serial doubling dilutions in assay medium are made down the plate to the penultimate row. δOμl of assay medium are added to all wells in the first column for each compound, and δOμl of HCMV strain Ad 169, diluted in medium to give a multiplicity of infection of 0.01 plaque-forming units per cell, are added to the wells in the remaining pairs of columns. Finally, 100μl of MRC-5 cell suspension at a concentration of 10⁵ cells per ml are added to each well. The plate is then incubated at 37°C in a humidified δ% C0₂ atmosphere for 7 days.

Detection of Viral Antigens (7d.p.i):

The growth medium is removed and the cell monolayers are washed once with phosphate buffered saline (PBS). The cells are fixed by the addition 1 :1 acetone:methanol for δ minutes and washed again with PBS. In order to inhibit subsequent non-specific binding of antibodies, 100μl of PBS containing 0.06% Tween 20 and 2% w/v skimmed milk powder (blocking buffer) are added to each well and the plate is incubated for 1 hour at 37°C . The blocking solution is removed and the cells are washed once with PBS/0.05% Tween 20 (ELISA wash). The primary, murine, monoclonal antibody, specific for HCMV glycoprotein B, is diluted to 2μg/ml in blocking buffer and δOμl are added to each well. Following incubation for 1.6 hours at 37°C, the primary antibody solution is removed and the plate was washed 3 times. Horseradish peroxidase labelled, rabbit anti-mouse, polyclonal antibody, pre-adsorbed on uninfected MRC-5 cells, is diluted 1/1 ,500 in blocking buffer and 50μl are added to each well. After further incubation at 37°C for 1.5 hours, the secondary antibody solution is removed and the plate is washed thoroughly δ times and dried. δOμl of orthophenylenediamine/peroxide substrate in urea buffer are added to each well, and colour is allowed to develop at room temperature. The reaction is 163

stopped by the addition of 2δμl per well of 20% H₂S0₄ and the plate is read spectrophotometrically at 490nm.

The 50% inhibitory concentration (IC₅₀) value of an active compound is calculated by regression analysis of the plot of concentration against percentage reduction in absorbance compared to drug-free virus controls. On completion of the ELISA, the plate is washed, dried and the cells stained with 20% carbol fuchsin. An assessment of the lowest in-assay cytotoxic concentration of each compound is made by microscopic examination of both uninfected and infected cell monolayers.

HSV

δ Essentially similar to the HCMV ELISA with the following modifications:

(a) HSV 1 strain SC16 and HSV 2 strain 186 are assayed in Vero cells.

(b) The culture stage is reduced to 2 days

(c) The primary murine MAb is specific for HSV, recognising type 1 and 2 strains equally. 0

VZV

Again similar to the HCMV assay, with the following modifications: (a) VZV strain G31 is used. 5 (b) The culture stage is reduced to 4 days,

(c) The primary murine Mab is VZV specific.

Cytotoxicity assay 0

4,000 Vero cells suspended in 75μl of DMEM medium supplemented with δ% FCS are seeded into each well of a 96-well microplate. The cells are allowed to settle and adhere for 1 hour at 37°C, then 75μl per well of freshly prepared doubling dilutions of the compound from δOOμM are added. Following 96 hours 3δ incubation at 37°C, 20μl of a δmg/ml solution of 3-(4,5-dimethylthiazol-2-yl)-2,δ- 164

diphenyl-tetrazolium bromide (MTT) in phosphate buffered saline are added to each well. After a further 2 hour incubation at 37 °C, the supernatants are aspirated from the wells and 12δμl of acidified isopropanol containing 0.6% SDS are added to each. The plate is maintained on a shaking incubator for 20 minutes, then read spectrophotometrically at 690nm. The mean optical densities of replicate test wells are expressed as percentages of cell control well values, and are then plotted against drug concentration to allow calculation of the 50% toxic dose (CCIDs-,).

In vitro FRET assay for inhibitors of HSV-1 & 2 serine proteases

HSV1 protease was amplified by PCR from HSV-1 strain KOS using the enzyme PWO (Boehringer Mannheim) to produce a scaffold protein. A modified protease was inserted into the expression vector pRSET (invitrogen) and high level expression was obtained in E.coli strain BLR (DE3) (Novagen). The tagged protease was purified by metal affinity chromatography, dialysed into a buffer containing sodium citrate to stimulate autocatalytic cleavage to remove the scaffold fragment and affinity tag to yield the protease catalytic domain. The HSV-2 protease was PCR amplified from strain 186 in a similar way as the HSV- 1 protease above.

The assay used for testing inhibitor compounds is a FRET (Fluorescence resonance energy transfer) assay using the artificial substrate 'dabcyl- DNAVEASSKAPLK-EDANS'. The inhibitor compounds are tested in a mixture containing 100mM HEPES pH 7.5, 1 mM EDTA and 0.8M sodium citrate. Test compounds dissolved in DMSO are tested at a final concentrations of 1 or O.δμM in 1 % DMSO. Compounds are pre-incubated with 0.5μM of HSV-1 or HSV-2 protease for 70min at 25°C followed by addition of substrate at 2δμM to start the reaction. The plate is monitored for 120min in a fluorescence reader from TECAN Instruments ('SPECTRAFLUOR PLUS'), using an excitation and emission wavelengths of 360nm and 53δnm respectively. Data analysis was performed using Activity Base. Results are expressed as % inhibition by comparison of fluorescence readings with those from a control protease sample to which is added DMSO containing no test compound. In vitro pNA assay of hCMV viral serine protease inhibitor activity

The hCMV serine protease used is a mutant of the 30K protease lacking the internal cleavage site (Ala142/Ala143) and which has been cloned in E.coli to produce active enzyme (hCMV δAla protease). IC₅₀ data for test compounds are determined both without preincubation and after preincubation of the enzyme with test inhibitor compound for 16 minutes in order to demonstrate time dependency. Test compounds are dissolved in DMSO, serially diluted and added at a range of concentrations (from 100μM - 0.195μM) to a reaction containing O.δμM CMV δAla protease, 100mM HEPES pH7.5, 0.2mM EDTA,

10mM NaCl, 1 mM DTT, and 30% glycerol. The reaction mixture is pre- incubated at 32°C for 0 minutes or 15 minutes prior to addition of 4mM oligopeptide substrate (RESYVKA-pNA), and then analysed at 32°C in a BIO- TEK Bio Kinetics Reader EL340L

The assay substrate is RESYVKA-pNA

- ₂

O 1

RESYVK-pNA -≡ . RESYVK ' ' !

N H

Me (λ max =280nm)

O NQ₂

RESYVK + pNA ≡ = RESYVK + 1

OH H₂N

Me (λ max =405nm)

The plate reader monitors production of pNA and calculates the reaction rates over 30 minutes. The rates are plotted against inhibitor concentration and IC^ results determined at 0 and 1 δ minutes preincubation. Rate constant data (K,) are obtained in a similar manner to IC₅₀ data without preincubation, with the exception that reaction rates are measured over 67.5 minutes, giving steady state rates. Curves are plotted of rate against inhibitor concentration and the K, is calculated using a tight binding inhibition equation. 156

169

CO ro

Claims

Claims

A compound of the general formula (I):

or a pharmaceutically acceptable derivative thereof, wherein:

R represents H or substituted or unsubstituted Cι-₃ alkyl;

R represents optionally substituted heteroaryl or fused heteroaryl, with one to four heteroatoms, or R₉CO;

R₂ represents -S0₂R₃;

R₃ represents Cι-₆alkyl, Ci-βalkenyl, optionally substituted three to seven member, preferably four to seven member, N-heterocycle containing up to two heteroatoms,

R₄ and R₅ independently represent H or a substituted or unsubstituted group selected from Ci-ealkyl optionally including one or more heteroatoms, Cι-₆hydroxyalkyl, Cι-₆alkoxy, C₂,₈alkenyl, Cι-₄alkylC₃- cycloalkyl (optionally containing from 1 to 4 heteroatoms), C₁- ₆alkylCORι₃ Cι-₆alkylCH₂ORι_3> Cι-₆alkylNHCORι₃, C₁-₆alkylC0₂Ri3, C₂- 166

₈alkenylC0₂Ri₃, Cι-₆alkylCONR₁₃Rι_4l C^ealkylNR^R , aryl, Ci- ₃alkylaryl, heteroaryl and d-₃alkyl heteroaryl;

Re and R₇ independently represent H or a substituted or unsubstituted δ group selected from Cι-₆alkyl optionally including one or more heteroatoms, Cι-₆hydroxyalkyl, Ci-βalkenyl and Ci-ealkylOCONRisRiβ;

R₉ represents H or a substituted or unsubstituted group selected from Cι-₆alkyl, Cι-₆alkenyl, C₃-₇cycloalkyl, C₆-ιofused cycloalkyl, C₆-ιofused 0 cycloalkenyl, heteroaryl or fused heteroaryl containing one to four heteroatoms, aryl, and arylCι-₃alkyl;

R₁₃ and R₁₄ independently represent H or a substituted or unsubstituted group selected from Cι-₆alkyl, C₃- cycloalkyl, C₂-₈alkenyl, 6 Ci-βhaloalkyl, Ci-₆alkylaryl, heteroaryl or Cι-₆alkylheteroaryl;

R₁₅ and Ri₆ independently represent H or a substituted or unsubstituted C_halky! which may, together with the nitrogen atom to which they are attached, form a ring optionally containing one further 0 heteroatom.

2. A compound according to claim 1 wherein R is Cι-₃alkyl

3. A compound according to claim 1 or claim 2 wherein R is methyl. 6

4. A compound according to any one of claims 1 to 3 wherein Ri is selected from

wherein R' may be the same or different and represent H, Cι-₃alkyl or Cι-₃hydroxyalkyl and R" may be the same or different and represent H, OH, -OCOCι-₃alkyl, -OCONHCι-₃alkyl, or may together form a double bond;

optionally substituted with one or more groups selected from halogen, hydroxy, Cι-₃alkyl, Cι-₃hydroxyalkyl, Cι-₆alkylNHCORn, Ci- ealkylOCORn, Cι-₆alkylOCONHRn d-ealkylNHSO^_.Rn, Ci- ealkylNHCONHRn, wherein Rn represents H, Cι-₃alkyl or Cι-₃haloalkyl.

A compound according to any one of claims 1 to 4 wherein Ri is selected from

■ CH₂OH, Q, = CH₂0H, CH₂NHC0Me, CH₂NHC0Me, CH₂CH₂0H, CH,, CH₂CH₂NHCOMe, CH₂NHS0₂CH2Ph, CH₂OCONH e, CH₂NHS0₂ e,

CH₂0C0NHEt, CH₂NHCONHEt, CH₂NHCONHEt, CH₂NHS0₂CF₃

CH₂NHS0₂Me

CH₂NHCO <ζ

t OCONHEt

6. A compound according to any one of claims 1 to 5 wherein R₃ represents an optionally substituted four to seven member N- heterocycle containing up to two heteroatoms.

7. A compound according to claim 6 wherein R₃ represents an optionally substituted three to seven member N-heterocycle containing up to two heteroatoms.

8. A compound according to any one of claims 1 to 7 wherein R₃ is selected from

9. A compound according to any one of claims 1 to 8 wherein R-ι and R5 independently represent optionally substituted Cι-₆alkyl, aryl or Ci- ₆alkylCONRi₃Rι₄ wherein R₁₃ and R independently represent H or a substituted or unsubstituted group selected from Cι- alkyl, aryl, arylCi- ₆alkyl and heteroarylCi-₆alkyl.

10. A compound according to claim 9 wherein at least one of R1₃ and R represents substituted or unsubstituted benzyl.

1 1. A compound according to claim 10 wherein the benzyl ring bears one or more substituents (which may be the same or different) selected from halogen, trihalomethyl, nitro and methoxy.

12. A compound according to claim 9 wherein at least one of R or R5 is aryl, bearing one or more substituents selected from cyano, Cι-₃alkoxy, Cι-₃alkylsulphonyl, acetamido or Cι-₆alkylNR₈Rιo wherein R₈ and R10 independently represent H, Cι- alkyl, Cι-₃alkenyl, Cι-₃haloalkyl, or together form a ring which may optionally include a further heteroatom.

13. A compound according to claim 9 wherein R represents methyl and R5 is selected from C₁-₆alkylCONR₂ιR₂₂ heteroarylCH₂NHCOCH₂₎ δ heteroarylNHCOCH_2> arylCH₂NHCOCH₂₎ 4-Me₂NCH₂C₆H₄CH₂, and

HOCH₂CH(OH)CH₂ wherein R₂ι and R₂₂ may be the same or different and are selected from H and substituted benzyl bearing one or more substituents (which may be the same or different) selected from halogen, trihalomethyl, nitro, and methoxy.

14. A compound according to any one of claims 1 to 12 in enantiomerically pure cis-trans form in which the hydrogens at the two ring fusion carbons are trans to one another and the hydrogen at the R- substituted carbon is cis to that at the adjacent ring fusion carbon.

15. A compound according to any one of claims 1 to 13 for use in human or veterinary medicine.

16. A compound according to claim 14 for use in the treatment of conditions caused by viruses of the herpes family.

17. A compound according to claim 14 or 15 for use in the treatment of

HSV, VZV or CMV infections.

18. The use of a compound according to any one of claims 1 to 16 or a physiologically acceptable salt or solvate thereof in the manufacture of a medicament for the treatment of conditions caused by a viral infection.

19. The use according to claim 17 wherein the viral infection is caused by a member of the herpes family of viruses.

20. The use according to claim 17 or claim 18 wherein the viral infection is caused by HSV1 , HSV2, VZV or hCMV.

21. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17 in admixture with one or more physiologically acceptable diluents or carriers.

22. A process for preparation of a pharmaceutical composition according to claim 20 comprising mixing a compound according to any one of claims 1 to 17 in with one or more physiologically acceptable diluents or carriers.

23. A process for preparation of a compound according to any one of claims 1 to 13 comprising condensation of a compound of formula (M'):

with R RδNH, wherein

Ri represents optionally substituted heteroaryl or fused heteroaryl, with one to four heteroatoms, or R₉CO;

R₄ and R₅ independently represent H or a substituted or unsubstituted group selected from

optionally including one or more heteroatoms, Cι-₆hydroxyalkyl, Cι-₆alkoxy, C₂-₈alkenyl, Cι-₄alkylC₃- ₇cycloalkyl (optionally containing from 1 to 4 heteroatoms), Ci-

₆alkylCOR₁₃ Cι-₆alkylCH₂OR_13> Cι-₆alkylNHCORι₃, C -₆alkylC0₂Rι₃, C₂- ₈alkenylC0₂Ri3, Cι-₆alkylCONRι₃Ru, Cι-₆alkylNRι₃Ru, aryl, Ci- ₃alkylaryl, heteroaryl and Cι-₃alkylheteroaryl;

Rg represents H or a substituted or unsubstituted group selected from

Cι-₆alkyl, Cι-₆alkenyl, C₃-₇cycloalkyl, C₆-ιofused cycloalkyl, C₆-ιofused cycloalkenyl, heteroaryl or fused heteroaryl containing one to four heteroatoms, aryl, and arylCι-₃alkyl;

R13 and Ru independently represent H or a substituted or unsubstituted group selected from Cι-₆alkyl, C₃-rcycloalkyl, C₂-₈alkenyl, Cι-₆haloalkyl, Cι-₆alkylaryl, heteroaryl or Cι-₆alkylheteroaryl.