Benzamide derivatives useful in the treatment of muscular disorders and pain and for controlling spasticity and tremors

Abstract

The present invention relates to a compound of formula I, or a pharmaceutically acceptable salt thereof: (I) wherein: n is 0 or 1; R1 is selected from H, alkyl and aralkyl, wherein said alkyl and aralkyl groups may be optionally substituted by one or more OH groups; X is a group selected from CC(CH.sub.2).sub.p; C(R.sup.5)C(R.sup.6)(CH.sub.2).sub.q; and C(R.sup.5)(R.sup.6)C(R.sup.7)(R.sup.8)(CH.sup.2)r-; where each of R.sup.5, R.sup.6, R.sup.7 and R.sup.8 is independently H or alkyl, and each of p, q and r is independently 1, 2, 3, 4 or 5; Y is a group selected from: CN; COOR.sup.2; CONR.sup.3R.sup.4; SO.sub.2NR.sup.9R.sup.10; NR.sup.12COR.sup.13; NR.sup.14SO.sub.2R.sup.15; and a heterocyclic group selected from oxadiazolyl, thiazolyl, iso-thiazolyl, oxazolyl, iso-oxazolyl, pyrazolyl and imidazolyl; where each of R.sup.2, R.sup.3 and R.sup.4 is independently H or alkyl; or R.sup.3 and R.sup.4 are linked, together with the nitrogen to which they are attached, to form a 5 or 6-membered heterocycloalkyl or heterocycloalkenyl group, said heterocycloalkyl or heterocycloalkenyl group optionally containing one or more further groups selected from O, N, CO and S, and where each of R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14 and R.sup.15 is independently H or alkyl. Further aspects of the invention relate to the use of such compounds in the preparation of a medicament for the treatment of a muscular disorder, pain, or for controlling spasticity or tremors, for example, spasticity in MS. ##STR00001##

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof, ##STR00146## wherein: n is 0 or 1; R.sup.1 is selected from H, alkyl and aralkyl, wherein said alkyl and aralkyl groups may be optionally substituted by one or more OH groups; X is a group selected from CC(CH2)p-; C(R.sup.5)C(R.sup.6)(CH.sub.2).sub.q; and C(R.sup.5)(R.sup.6)C(R.sup.7)(R.sup.8)(CH.sub.2).sub.r; where each of R.sup.5, R.sup.6, R.sup.7 and R.sup.8 is independently H or unsubstituted alkyl, and each of p, q, and r is independently 1, 2, 3, 4 or 5; Y is a group selected from: CN; COOR.sup.2; CONR.sup.3R.sup.4; SO.sub.2NR.sup.9R.sup.10; NR.sup.12COR.sup.13; NR.sup.14SO.sub.2R.sup.15; and a heterocyclic group selected from oxadiazolyl, thiazolyl, iso-thiazolyl, oxazolyl, iso-oxazolyl, pyrazolyl and imidazolyl; where each of R.sup.2, R.sup.3 and R.sup.4 is independently H or unsubstituted alkyl; or R.sup.3 and R.sup.4 are linked, together with the nitrogen to which they are attached, to form an unsubstituted 5 or 6-membered heterocycloalkyl or an unsubstituted heterocycloalkenyl group, said heterocycloalkyl or heterocycloalkenyl group optionally containing one or more further groups selected from O, N, CO and S, and where each of R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14 and R.sup.15 is independently H or unsubstituted alkyl.

2. A compound according to claim 1 wherein R1 is selected from H, Me, Et, .sup.nPr, .sup.iPr, CH.sub.2-phenyl, CH.sub.2-[4-(OH)-phenyl], CH.sub.2OH, CH(OH)CH.sub.3, CH(CH.sub.3)CH.sub.2CH.sub.3 and CH.sub.2CH(CH.sub.3).sub.2.

3. A compound according to claim 1 wherein Y is selected from CN, CON(Me).sub.2, CONHMe, CONHEt, SO.sub.2N(Me).sub.2, N(Me)COMe, N(Me)SO.sub.2Me, CO-piperidinyl, CO-pyrrolidinyl, oxadiazolyl and thiazolyl, more preferably, CON(Me).sub.2.

4. A compound according to claim 1 wherein X is cis C(R.sup.5)C(R.sup.6)(CH.sub.2).sub.q and q is 2, 3 or 4.

5. A compound according to claim 4 wherein X is CHCH(CH.sub.2).sub.q and q is 2 or 3.

6. A compound according to claim 1 wherein X is C(R.sup.5)(R.sup.6)C(R.sup.7)(R.sup.8)(CH.sub.2).sub.r and r is 2, 3 or 4.

7. A compound according to claim 6 wherein X is CH2-CH2-(CH2)r- and r is 2 or 3.

8. A compound according to claim 1 which is of formula Ia, or a pharmaceutically acceptable salt thereof, ##STR00147## wherein R1, R11, X, Y and n are as defined in claim 1.

9. A compound according to claim 1 which is of formula Ib, or a pharmaceutically acceptable salt thereof, ##STR00148## wherein R.sup.1, R.sup.11, X, Y and n are as defined in claim 1.

10. A compound according any claim 1 wherein n is 0.

11. A compound according any claim 1 wherein R1 is Me.

12. A compound according to claim 8 wherein n is 0, R1 is Me and X is CHCH(CH2)3- or CH2-CH2-(CH2)3-.

13. A compound according to claim 1 wherein n is 1 and R1 is H.

14. A compound which is selected from the following: ##STR00149## ##STR00150## ##STR00151## ##STR00152## ##STR00153## ##STR00154## ##STR00155## ##STR00156## ##STR00157## ##STR00158## ##STR00159## ##STR00160## and pharmaceutically acceptable salts, and enantiomers thereof.

15. A compound according to claim 1 which is of the formula [1], or a pharmaceutically acceptable salt thereof: ##STR00161##

16. A compound according to claim 1 which is of the formula [1a] or formula [1b], or a pharmaceutically acceptable salt thereof: ##STR00162##

17. A pharmaceutical composition comprising a compound according to claim 1 admixed with a pharmaceutically acceptable diluent, excipient or carrier.

18. A process for preparing a compound of formula I as defined in claim 1 which is of formula (VIII), wherein R.sup.1 and n are as defined in claim 1, said process comprising the steps of: ##STR00163## (i) coupling a compound of formula (V) with a compound of formula (III) to form a mixture comprising compounds of formulae (IIa) and (IIb); (ii) separating said compound of formula (IIb) from the mixture obtained in step (i); (iii) treating the compound of formula (IIb) obtained in step (ii) with a compound of formula (VI), where R.sup.1 and n are as defined in claim 1, and R is alkyl, to form a compound of formula (VII); and (iv) converting said compound of formula (VII) to a compound of formula (VIII).

19. A process for preparing a compound of formula I as defined in claim 1 which is of formula (VIII), wherein R.sup.1 and n are as defined in claim 1, said process comprising the steps of: ##STR00164## (i) treating a compound of formula (IX) with dimethylamine to form a compound of formula (X); (ii) reacting said compound of formula (X) with a compound of formula (XI) to form a compound of formula (XII); (iii) hydrogenating said compound of formula (XII) to form a compound of formula (IIb); (iv) treating said compound of formula (IIb) with a compound of formula (VI), where R.sup.1 and n are as defined in claim 1, and R is alkyl, to form a compound of formula (VII); and (v) converting said compound of formula (VII) to a compound of formula (VIII).

Description

(1) The present invention is further described by way of example, and with reference to the following figures wherein:

(2) FIG. 1 shows inhibition of electrically-evoked contractions of the mouse vas deferens by WIN55, VSN-22R and VSN-44.

(3) FIG. 2 shows the effect of VNS-44 (5 mg/kg i.v.) on spasticity (% spasticity vs baseline) against time after administration.

(4) FIG. 3 shows the dose response curve in mouse vas deferens for VSN-16R. The results show that the EC.sub.50 stays relatively constant across wt, CB.sub.1 knock-out and CB.sub.2 knock-out.

EXAMPLES

(5) General Procedures

(6) All starting materials and solvents were obtained either from commercial sources or prepared according to the literature citation. Unless otherwise stated all reactions were stirred.

(7) Normal phase column chromatography was routinely carried out on an automated flash chromatography system such as CombiFlash Companion or CombiFlash RF system. Intermediates were purified using pre-packed silica (230-400 mesh, 40-63 m) cartridges and products of a Lindlar reduction using pre-packed GraceResolv flash cartridges. SCX was purchased from Supelco or Silicycle (40-63 m size, 0.78 mmol/g loading).

(8) Analytical Methods

(9) Analytical HPLC was carried out using a Waters Xselect CSH C18, 2.5 m, 4.630 mm column eluting with a gradient of 0.1% Formic Acid in MeCN in 0.1% aqueous Formic Acid; a Waters Xbridge BEH C18, 2.5 m, 4.630 mm column eluting with a gradient of MeCN in aqueous 10 mM Ammonium Bicarbonate. UV spectra of the eluted peaks were measured using either a diode array or variable wavelength detector on an Agilent 1100 system.

(10) Analytical LCMS was carried out using a Waters Xselect CSH C18, 2.5 m, 4.630 mm column eluting with a gradient of 0.1% Formic Acid in MeCN in 0.1% aqueous Formic Acid; a Waters Xbridge BEH C18, 2.5 m, 4.630 mm column eluting with a gradient of MeCN in aqueous 10 mM Ammonium Bicarbonate. UV and mass spectra of the eluted peaks were measured using a variable wavelength detector on either an Agilent 1200 with or an Agilent Infinity 1260 LCMS with 6120 single quadrupole mass spectrometer with positive and negative ion electrospray.

(11) Preparative HPLC was carried out using a Waters Xselect CSH C18, 5 m, 1950 mm column using either a gradient of either 0.1% Formic Acid in MeCN in 0.1% aqueous Formic Acid or a gradient of MeCN in aqueous 10 mM Ammonium Bicarbonate; or a Waters Xbridge BEH C18, 5 m, 1950 mm column using a gradient MeCN in aqueous 10 mM Ammonium Bicarbonate; or the compounds were purified by reverse-phase HPLC (Gilson) using preparative C-18 column (Hypersil PEP 10021 mm internal diameter, 5 m particle size, and 100 pore size) and isocratic gradient over 20 minutes. Fractions were collected following detection by UV at a single wavelength measured by a variable wavelength detector on a Gilson 215 preparative HPLC or Varian PrepStar preparative HPLC; by mass and UV at a single wavelength measured by a ZQ single quadrupole mass spectrometer, with positive and negative ion electrospray, and a dual wavelength detector on a Waters FractionLynx LCMS.

(12) .sup.1H NMR Spectroscopy: .sup.1H NMR spectra were acquired on a Bruker Avance III spectrometer at 400 MHz. The central peak of dimethylsulfoxide-d.sub.6 was used as reference.

Abbreviations

(13) AcOH glacial acetic acid aq. aqueous br broad d doublet dd doublet of doublets ddd double double doublet dt doublet of triplets DCM dichloromethane DIPEA diisopropylethylamine DMF dimethylformamide DMSO dimethyl sulfoxide (ES+) electrospray ionization, positive mode Et ethyl Et.sub.3N triethylamine EtOAc ethyl acetate EtOH ethanol HATU 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HCl hydrochloric acid HPLC high performance liquid chromatography hr hour(s) Hz hertz LC liquid chromatography (M+H)+ protonated molecular ion M molar m multiplet Me methyl MeCN acetonitrile MeOH methanol MgSO.sub.4 magnesium sulphate MHz megahertz min minute(s) MS mass spectrometry m/z: mass-to-charge ratio Na.sub.2SO.sub.4 sodium sulphate NMR nuclear magnetic resonance (spectroscopy) Ph phenyl ppm parts per million q quartet qn quintet rt room temperature HPLC high performance liquid chromatography s singlet sat. saturated SCX solid supported cation exchange (resin) t triplet td triplet of doublets TEA triethylamine THF tetrahydrofuran TLC thin layer chromatography wt % weight percent Prefixes n-, s-, t- and tert- have their usual meanings: normal, secondary, iso, and tertiary.

(14) General Method for Amide Coupling:

(15) To a suspension of carboxylic acid (1.0 eq.), amine or amine.HCl salt (1.05-1.1 eq.) and HATU (1.1-1.3 eq.) in dry DCM (10 mL/g) was added DIPEA or TEA (2.0-3.0 eq.). The reaciton was stirred at it until complete by LCMS. The volatiles were removed in vacuo and the residue partitioned between EtOAc (20 mL/g) and sat. aq. ammonium chloride (20 mL/g). The aqueous layer was extracted with EtOAc (220 mL/g) before the combined organic extracts were washed with sat. aq. ammonium chloride (30 mL/g), water (30 mL/g) then brine (30 mL/g) and dried (MgSO.sub.4 or Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The crude material was purified by column chromatography.

(16) General Method for Sonogashira Coupling:

(17) To a solution of aryl iodide (1.0 eq.) and diisopropylamine (1.2 eq.) in dry THF (10 mL/g) under nitrogen was added bis(triphenylphosphine)palladium(II) chloride (4 mol %) and copper(I) iodide (7 mol %). The reaction was stirred for 5 min before alkyne (1.1-1.5 eq.) was added. The reaction was then heated at 60 C. for 1 h before the solvent was removed in vacuo and the residue partitioned between EtOAc (20 mL/g) and sat. aq. ammonium chloride (20 mL/g). The aq. layer extracted with EtOAc (220 mL/g) before the combined organic extracts were washed with sat. aq. ammonium chloride (20 mL/g), water (20 mL/g) and brine (20 mL/g) then dried (MgSO.sub.4), filtered and concentrated. The crude material purified by column chromatography to yield desired coupled product.

(18) General Method for Lindlar Reduction:

(19) To a flask containing palladium on barium sulphate reduced (5%) (50 wt % cf. alkyne) under nitrogen was added a solution of alkyne (1.0 eq.) and quinoline (1.3 eq.) in MeOH (40 mL/g). The vessel was placed under an atmosphere of hydrogen until the reaction was deemed complete by TLC, HPLC or LCMS analysis. The catalyst was removed by filtration through celite and the quinoline was removed by filtration through SCX (washing several times with MeOH). The filtrate

(20) General Method for Ester Saponification:

(21) To a solution of ester (1.0 eq.) in THF (10 mL/g) was added a solution of lithium hydroxide (1.5-2.0 eq.) in water (1 mL/g). The reaction was stirred at rt until judged complete by HPLC or LCMS analysis. The volatiles were removed in vacuo and the residue was partitioned with EtOAc (10 mL/g). The aqueous layer was acidified to pH 1 with 1 N citric acid and extracted with EtOAc (310 mL/g). The combined organic extracts were washed with water (210 mL/g) and brine (10 mL/g) then dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo.

(22) General Procedure for Reduction of Alkyne to Alkane:

(23) To a flask containing alkyne (1.0 eq.) in EtOH (15-20 mL/g) under nitrogen was added palladium on carbon (5 wt %) (50 wt % cf. alkyne). The mixture was placed under an atmosphere of hydrogen (2 bar) until judged complete by LCMS analysis. The catalyst was removed by filtration through celite and washed well with EtOH. The filtrate was then concentrated in vacuo and purified by chromatography to give the desired alkane product.

(24) Preparation of VSN-44

(25) The compound VSN-44 can be prepared by the following methodology. Other compounds of formula I can be prepared by analogous methodology using commercially availably starting materials and standard synthetic steps that would be familiar to the skilled artisan, including those set forth in WO 2005/080316 and WO 2010/116116.

3-[(1Z)-6-(dimethylamino)-6-oxohex-1-en-1-yl] benzoic acid (IIb)

(26) ##STR00025##

N,N-dimethylamino 4-(carboxybutyl)triphenylphosphonium bromide (III)

(27) 4-(carboxybutyl)triphenylphosphonium bromide (140 g, 0.315 mol, 1 equiv) was charged in a reactor and dichloromethane (650 ml, 4.5 vols) was added. Triethylamine (dried on molecular sieves; 95 ml, 2.1 equiv) was charged and the reaction mixture was cooled down to 10 C. Ethyl chloroformate (40 ml, 1.05 equiv) was added dropwise and the mixture was stirred for another 15 min at 10 C.

(28) A solution containing dimethylamine hydrochloride (freshly crystallised from methanol/ether; 78 g, 3 equiv) and triethylamine (200 ml, 4.5 equiv) in dichloromethane (1000 ml, 7 vols) was prepared.

(29) This solution was stirred for 40 min at room temperature and added dropwise to the reaction mixture at 10 C. The temperature was kept between 10 and 15 C. during all the addition. The reaction was left to warm up to room temperature. The reaction was stirred at room temperature overnight. The mixture was treated with 2 l of saturated NaHCO.sub.3 solution. The aqueous phase was extracted with dichloromethane (12 l and 21 l). Organics were combined and dried over MgSO.sub.4 and filtered. The volatiles were removed under vacuum. The residue was triturated with 350 ml of diethyl ether. The solid was filtered and triturated with hot diethyl ether for 5 hours. The suspension was cooled down and the solid filtered. The solid was dried under vacuum to give 130.9 g of a white solid (III) (90% yield).

(30) .sup.1H NMR (CDCl.sub.3) 7.65-8.0 (m, 15H); 3.7 (m, 2H); 3.0 (s, 3H); 2.8 (s, 3H), 2.5 (t, J=7 Hz, 2H); 1.9 (m, 2H), 1.7 (m, 2H).

3-[(1Z)-6-(dimethylamino)-6-oxohex-1-en-1-yl] benzoic acid (IIb)

(31) N,N-dimethylamino 4-carboxybutyltriphenylphosphonium (III) (61.9 g, 0.13 mol, 3 equivalents) were dissolved in dry dichloromethane (150 ml, 2.4 vols) under nitrogen. The solution was cooled down to 0 C. and potassium hexamethyldisilazide (0.9M in THF; 45 ml, 5 equiv) was added dropwise at 0 C. The reaction mixture was stirred at 0 C. for another 45 min. A solution of methyl 3-formylbenzoate (7.16 g, 1 equiv) in dry THF (36 ml, 5 vols) was added keeping the temperature <4 C. The mixture was allowed to warm up to room temperature and was stirred for 18 hrs. The reaction was quenched with 2M HCl (400 ml) and extracted with dichloromethane (2400 ml and 2200 ml). Organics were combined, dried over MgSO.sub.4, filtered and evaporated to dryness. The residue was dissolved in a mixture of sodium hydroxide 1 M/methanol 4:1 (440 ml) and stirred for 18 hrs. Water (100 ml) was added to the mixture and methanol was evaporated under vacuum. Aqueous was extracted with ethyl acetate (400 ml). The pH was adjusted to pH 1 and the mixture was extracted with dichloromethane (2400 ml and 2200 ml). Organics were dried over MgSO.sub.4, filtered and evaporated to dryness. M=22.0 g. The crude was purified by flash chromatography using dichloromethane to dichloromethane/MeOH=95/54 as eluent. M=10.6 g 93% yield.

(32) Isomer Separation

(33) Acid (10.93 g, 0.042 mol) was dissolved in ethyl acetate (20 ml) and 4-dimethylaminopyridine (6.13 g, 1.2 equiv) was dissolved in warm ethyl acetate (20 ml). The DMAP solution was added to the free acid solution. The mixture was stirred at reflux temperature for 10 min. Then, the solution was allowed to cool down to room temperature slowly. A brown salt was formed, which was removed by filtration.

(34) A mixture of diethyl ether/ethyl acetate: 9:1 (40 ml) was added and the solution was heated to reflux. The mixture was stirred and allowed to cool down overnight. A pale yellow solid was filtered and dried in-vacuo. This solid was treated with HCl (1M) and extracted with dichloromethane (350 ml). Organics were dried over MgSO.sub.4, filtered and evaporated to dryness to give a brown oil which solidified upon standing (IIb). M=3.88 g (35.5% yield).

(35) 1H NMR (CDCl.sub.3) 9.7 (bs, 1H); 8.0 (m, 2H); 7.5 (m, 2H); 6.5 (d, J=11 Hz, 1H); 5.75 (m, 1H); 3.0 (s, 6H); 2.4 (m, 4H); 1.9 (m, 2H)

(36) Preparation of VSN44

(37) ##STR00026##

(38) To the substituted benzoic acid (IIb) (139 mg, 1 mmol) in DMF (1 mL) was added the Ala(OMe) in DMF (1 mL) and the PyBOP (572 mg, 1.1 mmol) added in DMF (2 mL). DIPEA (142 mg, 191 L, 1.1 mmol) was added dropwise, and the reaction stirred at room temperature overnight. Water (50 mL) was added and ethyl acetate (100 mL). The layers were stirred (5 mins), separated, and the ethyl acetate layer washed with brine (2100 mL), dried (Na.sub.2SO.sub.4) to give the crude product (650 mg). This was flash chromatographed using a 25 g Puriflash (silica) column, cyclohexane:acetone 15-45% gradient. Yield (IV) 180 mg, 0.54 mmol, 54%.

(39) 1H NMR (500 MHz, CDCl3) 7.77 (s, 1H), 7.71 (dt, J=1.6, 7.4, 1H), 7.42-7.38 (m, J=7.4, 1H), 7.38-7.31 (m, 2H), 6.46 (d, J=11.6, 1H), 5.74 (dt, J=7.7, 11.6, 1H), 4.84-4.76 (m, J=7.2, 1H), 3.77 (s, 3H), 2.95 (s, 3H), 2.90 (s, 3H), 2.42-2.30 (m, 4H), 1.83 (p, J=7.2, 2H), 1.64 (s, 2H), 1.54 (d, J=7.2, 3H).

(40) 13C NMR (126 MHz, CDCl3) 173.78, 172.63, 167.13, 137.88, 134.10, 133.23, 132.09, 128.90, 128.53, 127.12, 125.77, 52.54, 48.67, 37.30, 35.54, 32.60, 28.30, 24.99, 18.31, 17.66.

(41) Other compounds of formula I may be prepared by substituting Ala(OMe) in the above process with other commercially available amino acid esters.

(Z)-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzoyl)-D-alanine [1] (VSN-44)

(42) ##STR00027##

(43) The ester (IV) (135 mg, 0.41 mmol) in THF (2 mL) was added to lithium hydroxide, hydrate 84 mg, 2 mmol) in water (1 mL). The reaction was stirred at room temperature for 24 hrs. The THF was removed on the rotary evaporator and the residue taken up in 10% aq. Citric acid (10 mL). The aqueous mixture was extracted with DCM (330 mL) and dried over Na.sub.2SO.sub.4. Crude yield 307 mg. The product was purified by preparative LCMS (C18) using: Solvent A, 5% MeOH/95% H2O, 0.1% HCOOH. Solvent B, 95% MeOH/5% H.sub.2O, 0.1% HCOOH. Gradient 10% A to 95% over 8 min. The fractions were combined, and the volatiles removed on a rotary evaporator. The final aqueous mixture was freeze dried.

(44) 1H NMR (500 MHz, CDCl3) 9.03 (s, 1H), 7.74 (s, 1H), 7.73-7.67 (m, J=7.7, 2H), 7.38-7.34 (m, 1H), 7.34-7.31 (m, 1H), 6.43 (d, J=11.6, 1H), 5.70 (dt, J=7.7, 11.6, 1H), 4.80-4.70 (m, 1H), 2.96 (s, 3H), 2.89 (s, 3H), 2.35 (t, J=7.1, 3H), 2.32-2.22 (m, 1H), 1.86-1.73 (m, 2H), 1.54 (d, J=7.2, 3H).

(45) 13C NMR (126 MHz, CDCl3) 175.39, 173.51, 168.12, 137.75, 133.65, 132.95, 132.31, 128.96, 128.62, 127.07, 126.06, 49.28, 37.55, 35.84, 32.64, 28.27, 25.08, 17.84.

(46) Alternative Synthesis of Intermediate (IIb)

(47) (i) Stage 1

(48) ##STR00028##

(49) 5-Hexynoic acid (553 g, 4.91 mol) and dichloromethane (5.5 L, 10 vol) are charged to a 10 L vessel and cooled to 7 C. Oxalyl chloride (0.475 L, 5.40 mol) is added dropwise maintaining the temperature between 4.5 and 5.0 C. over a 2 h period. The addition apparatus was washed with dichloromethane and stirred for 10 mins at 5 C. Dimethylformamide was added portion-wise with mild effervescence. The temperature was taken to 2 C. and the mixture stirred for 2 h and then warmed to 12 C. and stirred for a further 16 h until no further discernable reaction was observed. The mixture was concentrated to remove all oxalyl chloride. The vessel was rinsed with dichloromethane. Dimethylamine hydrochloride (490 g, 5.89 mol) and dichloromethane (5.5 L, 10 vol) were charged to the 10 L vessel. Triethylamine (2.5 L, 15.70 mol) was charged and the mixture cooled to 10 C. The concentrated acid chloride was treated with dichloromethane (0.3 L, 0.55 vol) and added dropwise maintaining temperature below 6 C. The addition apparatus was rinsed with dichloromethane (50 ml, 0.1 vol) the mixture was stirred at 5 C. for 15 mins and then allowed to warm to ambient temperature. When no further discernable reaction was observed. Water (3 L, 5.5 vol) was charged stirred and the layers partitioned. The aqueous was washed with dichloromethane (2.5 L, 4.5 vol). The organic layers were combined and then washed with 2M Hydrochloric acid (2.51, 4.5 vol), 1M NaOH (2.5 L, 4.5 vol), water (3 L, 5.4 vol), brine (2.5 L, 4.5 vol) and dried over MgSO.sub.4 (100 g, 20 wt %). The suspension was filtered and the solvent removed to give a dark oil (X) (214 g, 83%) GF1218-47-128 (568 g, 83%)

(50) (ii) Stage 2

(51) ##STR00029##

(52) 3-Bromobenzoic acid (XI) (631 g, 3.14 mol, 1.0 eq) and piperidine (1.55 L,) were charged to the vessel leading to a mild exotherm and the mixture was heated to 85 C. Dichlorobis(triphenylphosphine)Palladium (II) (44 g, 0.06 mol) was charged, followed by slow addition of N,N-dimethylhex-5-ynamide (X) (656 g, 4.71 mol) maintaining the temperature below 116 C. (reflux). The reaction was stirred for a further 1 hour until no further reaction was observed and allowed to cool to ambient temperature. The resulting viscous mixture was dissolved in water (9 L) an acidified with 5M HCl (4 L) and then extracted with ethyl acetate (5.5, 3.5 and 3 L). The organics were combined and washed with water (3 L) and brine (2 L) and then the solvent removed to give a dark oil. The material was taken in acetonitrile (2.5 L) and passed through silica(1.5 Kg) washing with acetonitrile (2.5 L). The resulting solution crystallised and the solid was collected (100 g). The liquors(4 L) were concentrated and crystallised to give the desired product as a solid (49 g). The silica was eluted with ethyl acetate (2 L), which yielded further product (54 g) after concentration. A further three portions of ethyl acetate (2 L) were used as eluent to give further product (40 g, 20 g and 17 g) respectively. The fractions were combined and treated with acetonitrile (340 ml) and recrystallised from the same solvent to give a pale yellow solid (XII) (105 g).

(53) (iii) Stage 3

(54) ##STR00030##

(55) The alkyne (XII) (105 g, 0.4 mol, 1.0 eq) and 5% Pd on BaSO.sub.4 (5.25 g, 5 wt %), methanol (25 vol) and quinoline (3.68 ml, 0.035 vol) were charged to the vessel The vessel was evacuated and the atmosphere replaced with hydrogen three times and then left to react at room temperature under a positive pressure of hydrogen until no further starting material was observable. The solution was degassed and the atmosphere replaced with nitrogen. The suspension was filtered through cellite and washed with methanol (1 L). The solution was then concentrated to dryness and taken in ethyl acetate (5 vol) and washed with 2M HCl (32 vol) and brine (3 vol).The solvent was removed and the resulting oil was taken up in acetone (3.3 vol) stirred and cooled until crystallisation occurred. The product was filtered and washed with cold acetone (0.5 vol) to give a colourless solid (IIb) (111 g, 61%).

(56) Synthesis of VSN 45-47

(R)-methyl 2-(3-iodobenzamido)propanoate

(57) ##STR00031##

(58) Using the general procedure described for amide coupling, the reaction of 3-iodobenzoic acid (22.3 g, 90 mmol), (R)-methyl 2-aminopropanoate.HCl (13.55 g, 97 mmol), HATU (37.6 g, 99 mmol) and TEA (31.3 ml, 225 mmol) in dry DCM (200 mL) gave the title compound (R)-methyl 2-(3-iodobenzamido)propanoate (41 g, 96% yield) as a pale yellow oil. No purification required.

(59) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 3.64 (3H, s), 4.47 (1H, qn, J=7.3 Hz), 7.30 (1H, t, J=7.8 Hz), 7.89 (1H, ddd, J=1.1, 1.6, 7.8 Hz), 7.93 (1H, ddd, J=1.0, 1.7, 7.8 Hz), 8.25 (1H, t, J=1.6 Hz), 8.92 (1H, d, J=6.8 Hz) ppm. MS(ES+) m/z 334.0 (M+H).

(R)-6-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)hex-5-ynoic acid

(60) ##STR00032##

(61) Following the general method for Sonogashira coupling, the reaction of (R)-methyl 2-(3-iodobenzamido)propanoate (15.0 g, 36.0 mmol) and hex-5-ynoic acid (4.57 ml, 41.4 mmol) after purification by column chromatography (1-3% MeOH in DCM) gave (R)-6-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)hex-5-ynoic acid (8.43 g, 69.3% yield).

(62) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.78 (2H, qn, J=7.2 Hz), 2.39 (2H, t, J=7.3 Hz), 2.46-2.49 (2H, m), 3.64 (3H, s), 4.47 (1H, qn, J=7.3 Hz), 7.46 (1H, t, J=7.8 Hz), 7.56 (1H, td, J=1.3, 7.7 Hz), 7.82 (1H, td, J=1.4, 7.8 Hz), 7.92 (1H, t, J=1.5 Hz), 8.87 (1H, d, J=6.9 Hz), 12.16 (1H, s) ppm.

(63) MS(ES+) m/z 318 (M+H).

(R)-methyl 2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-yn-1-yl)benzamido)propanoate VSN 45

(64) ##STR00033##

(65) Using the general procedure described for amide coupling, the reaction of (R)-6-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)hex-5-ynoic acid (0.80 g, 2.52 mmol), pyrrolidine (0.22 ml, 2.65 mmol), DIPEA (1.35 ml, 7.56 mmol), HATU (1.15 g, 3.03 mmol) and dry DCM (10 mL) after purification by chromatography (1-4% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-yn-1-yl)benzamido)propanoate (0.6 g, 63.0% yield) as a pale yellow oil.

(66) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.69-1.93 (6H, m), 2.39 (2H, t, J=7.2 Hz), 2.44-2.52 (2H, m), 3.28 (2H, t, J=6.8 Hz), 3.41 (2H, t, J=6.8 Hz), 3.64 (3H, s), 4.47 (1H, qn, J=7.3 Hz), 7.46 (1H, t, J=7.7 Hz), 7.56 (1H, td, J=1.3, 7.7 Hz), 7.82 (1H, td, J=1.5, 7.8 Hz), 7.92 (1H, t, J=1.5 Hz), 8.87 (1H, d, J=6.9 Hz) ppm.

(67) (.sup.13C) DMSO-d.sub.6: 16.65, 18.23, 32.56, 38.22, 45.21, 45.84, 48.28, 51.89, 80.20, 91.12, 123.20, 127.08, 128.74, 130.03, 133.91, 133.98, 165.41, 169.67, 173.05 ppm.

(68) MS(ES+) m/z 371 (M+H).

(R,Z)-methyl 2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-en-1-yl)benzamido)propanoate VSN 46

(69) ##STR00034##

(70) Following the general procedure for Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-yn-1-yl)benzamido)propanoate (0.5 g, 1.350 mmol) gave the named product with trace amounts of the trans double bond isomer and fully saturated products (determined by .sup.1H NMR). Separation by column chromatography (1-3% MeOH in DCM) gave the title compound (0.17 g, 33.1%). The other 2 components were not isolated.

(71) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.60-1.76 (4H, m), 1.82 (2H, qn, J=6.6 Hz), 2.25 (2H, t, J=7.2 Hz), 2.31 (2H, dq, J=1.6, 7.5 Hz), 3.22 (2H, t, J=6.9 Hz), 3.31-3.36 (2H, m), 3.64 (3H, s), 4.48 (1H, qn, J=7.3 Hz), 5.74 (1H, td, J=7.3, 11.7 Hz), 6.48 (1H, d, J=11.7 Hz), 7.42-7.49 (2H, m), 7.70-7.80 (2H, m), 8.81 (1H, d, J=6.9 Hz) ppm.

(72) (.sup.13C) DMSO-d.sub.6: 16.70, 23.89, 24.44, 25.55, 27.66, 33.03, 45.15, 45.79, 48.25, 51.86, 125.69, 127.47, 128.25, 128.36, 131.35, 133.30, 133.74, 137.11, 166.14, 169.98, 173.12 ppm.

(73) MS(ES+) m/z 373 (M+H).

(R,Z)-2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-en-1-yl)benzamido)propanoic acid VSN 47

(74) ##STR00035##

(75) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-en-1-yl)benzamido)propanoate (0.15 g, 0.40 mmol) with lithium hydroxide (19 mg, 0.81 mmol) gave (R,Z)-2-(3-(6-oxo-6-(pyrrolidin-1-yl)hex-1-en-1-yl)benzamido)propanoic acid (0.13 g, 88% yield) as a white solid.

(76) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.4 Hz), 1.68 (4H, m), 1.81 (2H, qn, J=6.6 Hz), 2.24 (2H, t, J=7.2 Hz), 2.27-2.37 (2H, m), 3.22 (2H, t, J=6.8 Hz), 3.32 (2H, t, J=6.8 Hz), 24.42 (1H, qn, J=7.3 Hz), 5.74 (1H, td, J=7.3, 11.7 Hz), 6.48 (1H, d, J=11.7 Hz), 7.41-7.50 (2H, m), 7.71-7.81 (2H, m), 8.71 (1H, d, J=7.3 Hz), 12.54 (1H, s) ppm.

(77) (.sup.13C) DMSO-d.sub.6: 16.88, 23.94, 24.49, 25.59, 27.70, 33.07, 45.20, 45.83, 48.17, 125.71, 127.52, 128.25, 128.43, 131.28, 133.30, 134.00, 137.10, 166.06, 170.02, 174.23 ppm.

(78) MS(ES+) m/z 359 (M+H).

(79) Synthesis of VSN 48-50

6-(3-(methoxycarbonyl)phenyl)hex-5-ynoic acid

(80) ##STR00036##

(81) Following the general method for Sonogashira coupling, the reaction of methyl 3-iodobenzoate (1.0 g, 3.82 mmol) and hex-5-ynoic acid (0.421 ml, 3.82 mmol) after purification by column chromatography (0-3% MeOH in DCM) gave 6-(3-(methoxycarbonyl)phenyl)hex-5-ynoic acid (0.78 g, 81% yield).

(82) (.sup.1H) DMSO-d.sub.6: 1.79 (2H, qn, J=7.2 Hz), 2.39 (2H, t, J=7.3 Hz), 2.45-2.49 (2H, m), 3.86 (3H, s), 7.47-7.55 (1H, m), 7.66 (1H, td, J=1.4, 7.7 Hz), 7.88-7.94 (2H, m), 12.07 (1H, s) ppm.

(83) MS(ES+) m/z 247 (M+H).

Methyl 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoate

(84) ##STR00037##

(85) Following the general procedure described for amide coupling, the reaction 6-(3-(methoxycarbonyl)phenyl)hex-5-ynoic acid (5.2 g, 21.12 mmol), dimethylamine.HCl (2.07 g, 25.3 mmol), DIPEA (11.28 ml, 63.3 mmol) and HATU (10.4 g, 27.5 mmol) in dry DCM (50 mL) gave methyl 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoate (5.3 g, 87% yield) as an orange oil. No purification required.

(86) (.sup.1H) DMSO-d.sub.6: 1.77 (2H, qn, J=7.2 Hz), 2.36-2.49 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.86 (3H, s), 7.47-7.54 (1H, m), 7.66 (1H, td, J=1.4, 7.7 Hz), 7.89-7.91 (2H, m) ppm.

(87) MS(ES+) m/z 274 (M+H).

3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid

(88) ##STR00038##

(89) To a solution of methyl 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoate (5.30 g, 18.42 mmol) in THF (40 mL) and water (20 mL) was added lithium hydroxide (0.88 g, 36.8 mmol). The reaction was stirred at rt until judged complete by HPLC analysis. The volatiles were then removed in vacuo and the residue was partitioned between water (60 mL) and EtOAc (50 mL). The aqueous layer was then acidified to pH 1 with 1 N HCl (aq) and extracted with EtOAc (3100 mL). Next, the combined organic extracts were washed with water (275 mL) and brine (50 mL) then dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to a residue. This was azeotroped with iso-hexanes and dried in vacuum desiccator (45 C.) to give 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (4.3 g, 88% yield) as an orange solid.

(90) (.sup.1H) DMSO-d.sub.6: 1.78 (2H, qn, J=7.2 Hz), 2.41-2.49 (4H, m), 2.83 (3H, s), 2.98 (3H, s), 7.46-7.52 (1H, m), 7.64 (1H, td, J=1.5, 7.7 Hz), 7.87-7.91 (2H, m), 13.11 (1H, s) ppm.

(91) MS(ES+) m/z 260 (M+H).

methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)acetate VSN 48

(92) ##STR00039##

(93) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.70 g, 2.65 mmol), methyl 2-aminoacetate.HCl (0.38 g, 3.04 mmol), DIPEA (1.4 ml, 7.94 mmol) and HATU (1.31 g, 3.44 mmol) in dry DCM (10 mL) after purification by chromatography (1-3% MeOH in DCM) gave the title compound methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)acetate (0.86 g, 93% yield) as a pale yellow oil.

(94) (.sup.1H) DMSO-d.sub.6: 1.77 (2H, qn, J=7.2 Hz), 2.42-2.48 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.65 (3H, s), 4.01 (2H, d, J=5.8 Hz), 7.47 (1H, t, J=7.8 Hz), 7.57 (1H, td, J=1.3, 7.7 Hz), 7.81 (1H, td, J=1.4, 7.8 Hz), 7.89 (1H, t, J=1.5 Hz), 9.04 (1H, t, J=5.8 Hz) ppm.

(95) (.sup.13C) DMSO-d.sub.6: 18.22, 23.92, 31.18, 34.77, 36.63, 38.22, 41.19, 51.75, 80.14, 91.18, 123.33, 126.85, 128.85, 129.92, 133.86 and 134.05, 165.81, 170.25, 171.30 ppm.

(96) MS(ES+) m/z 331 (M+H).

(Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)acetate VSN 49

(97) ##STR00040##

(98) Following the general procedure for the Lindlar reduction, the hydrogenation of methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)acetate (0.40 g, 1.21 mmol) gave the named product with trace amounts of the trans double bond isomer and fully saturated products (determined by .sup.1H NMR). Separation by column chromatography (1-3% MeOH in DCM) gave the title compound (0.21 g, 51.1% yield). The other 2 components were not isolated.

(99) (.sup.1H) DMSO-d.sub.6: 1.65 (2H, qn, J=7.3 Hz), 2.27-2.36 (4H, m), 2.77 (3H, s), 2.92 (3H, s), 3.66 (3H, s), 4.02 (2H, d, J=5.9 Hz), 5.75 (1H, dt, J=7.4, 11.7 Hz), 6.47 (1H, br d, J=11.7 Hz), 7.47 (2H, dd, J=1.2, 4.0 Hz), 7.70-7.81 (2H, m), 8.98 (1H, t, J=5.8 Hz) ppm.

(100) (.sup.13C) DMSO-d.sub.6: 24.73, 27.73, 31.69, 34.74, 36.62, 41.23, 51.72, 125.53, 127.21, 128.29, 128.40, 131.50, 133.34, 133.65, 137.20, 166.50, 170.35, 171.65 ppm.

(101) MS(ES+) m/z 333 (M+H).

(Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)acetic acid VSN 50

(102) ##STR00041##

(103) Following the general procedure for saponification, the reaction of (Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)acetate (0.1 g, 0.30 mmol) with lithium hydroxide (14 mg, 0.60 mmol) gave (Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)acetic acid (76 mg, 77% yield) as a colourless gum.

(104) (.sup.1H) DMSO-d.sub.6: 1.66 (2H, qn, J=7.3 Hz), 2.25-2.38 (4H, m), 2.78 (3H, s), 2.92 (3H, s), 3.94 (2H, d, J=5.9 Hz), 5.75 (1H, dt, J=7.4, 11.7 Hz), 6.48 (1H, d, J=11.8 Hz), 7.47 (2H, dd, J=1.5, 3.9 Hz), 7.71-7.77 (1H, m), 7.79 (1H, s), 8.87 (1H, t, J=5.8 Hz), 12.56 (1H, s) ppm.

(105) (.sup.13C) DMSO-d.sub.6: 24.74, 27.74, 31.71, 34.75, 36.63, 41.23, 125.52, 127.21, 128.33, 128.36, 131.39, 133.29, 133.88, 137.16, 166.37, 171.29, 171.66 ppm.

(106) MS(ES+) m/z 319 (M+H).

(107) Synthesis of VSN 51 to 53

(108) (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate VSN 51

(109) ##STR00042##

(110) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.70 g, 2.65 mmol), (S)-methyl 2-amino-3-hydroxypropanoate.HCl (0.45 g, 2.91 mmol), DIPEA (1.18 ml, 6.61 mmol) and HATU (1.16 g, 3.04 mmol) in dry DCM (10 mL) after purification by chromatography (1-5% MeOH in DCM) gave the title compound (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate (0.67 g, 68.9% yield) as a pale yellow oil.

(111) (.sup.1H) DMSO-d.sub.6: 1.78 (2H, qn, J=7.2 Hz), 2.42-2.49 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.65 (3H, s), 3.79 (2H, t, J=5.9 Hz), 4.52 (1H, dt, J=5.5, 7.4 Hz), 5.04 (1H, t, J=6.2 Hz), 7.47 (1H, t, J=7.7 Hz), 7.57 (1H, td, J=1.3, 7.7 Hz), 7.83 (1H, td, J=1.5, 7.8 Hz), 7.93 (1H, t, J=1.5 Hz), 8.68 (1H, d, J=7.4 Hz) ppm.

(112) (.sup.13C) DMSO-d.sub.6: 18.23, 23.93, 31.20, 34.78, 36.63, 51.89, 55.71, 60.94, 80.18, 91.16, 123.21, 127.12, 128.76, 130.07, 133.98, 134.04, 165.73, 170.92, 171.30 ppm.

(113) MS(ES+) m/z 361 (M+H).

(S,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoate VSN 52

(114) ##STR00043##

(115) Following the general procedure for the Lindlar reduction, the hydrogenation of (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate (0.50 g, 1.39 mmol) gave the named product along with the trans double bond isomer (10%) and fully saturated product (20%) (determined by .sup.1H NMR). Separation by column chromatography (1-3% MeOH in DCM) gave the title compound (0.28 g, 54.6% yield). The other 2 components were not isolated.

(116) (.sup.1H) DMSO-d.sub.6: 1.65 (2H, qn, J=7.4 Hz), 2.26-2.35 (4H, m), 2.77 (3H, s), 2.91 (3H, s), 3.65 (3H, s), 3.79 (2H, t, J=5.8 Hz), 4.54 (1H, dt, J=5.4, 7.4 Hz), 5.04 (1H, t, J=6.2 Hz), 5.75 (1H, dt, J=7.3, 11.7 Hz), 6.48 (1H, d, J=11.7 Hz), 7.44-7.50 (2H, m), 7.71-7.81 (2H, m), 8.60 (1H, d, J=7.4 Hz) ppm.

(117) (.sup.13C) DMSO-d.sub.6: 24.79, 27.71, 31.72, 34.74, 36.62, 51.86, 55.66, 60.99, 125.67, 127.57, 128.29, 128.33, 131.36, 133.33, 133.81, 137.15, 166.45, 171.03, 171.61 ppm.

(118) MS(ES+) m/z 363 (M+H).

(S,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoic acid VSN 53

(119) ##STR00044##

(120) Following the general procedure for saponification, the reaction of (S,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoate (0.15 g, 0.42 mmol) with lithium hydroxide (20 mg, 0.83 mmol) gave (S,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoic acid (75 mg, 51.0% yield) as a colourless gum.

(121) (.sup.1H) DMSO-d.sub.6: 1.65 (2H, qn, J=7.4 Hz), 2.18-2.41 (4H, m), 2.77 (3H, s), 2.91 (3H, s), 3.80 (2H, d, J=5.2 Hz), 4.48 (1H, dt, J=7.7, 5.2 Hz), 4.95 (1H, br s), 5.75 (1H, dt, J=7.3, 11.7 Hz), 6.49 (1H, d, J=11.8 Hz), 7.47 (2H, d, J=5.0 Hz), 7.66-7.86 (2H, m), 8.42 (1H, d, J=7.7 Hz), 12.67 (1H, s) ppm.

(122) (.sup.13C) DMSO-d.sub.6: 24.80, 27.72, 31.73, 34.75, 36.63, 55.66, 61.16, 125.61, 127.53, 128.26, 128.37, 131.25, 133.29, 134.07, 137.13, 166.32, 171.62, 171.90 ppm.

(123) MS(ES+) m/z 349 (M+H).

(124) Synthesis of VSN 54 to 56

(S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate VSN 54

(125) ##STR00045##

(126) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.70 g, 2.65 mmol), (S)-methyl 2-amino-3-phenylpropanoate.HCl (0.60 g, 2.78 mmol), DIPEA (1.18 ml, 6.61 mmol) and HATU (1.16 g, 3.04 mmol) in dry DCM (10 mL) after purification by chromatography (1-4% MeOH in DCM) gave the title compound (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate (0.83 g, 72.4% yield) as a pale yellow oil.

(127) (.sup.1H) DMSO-d.sub.6: 1.71-1.84 (2H, m), 2.42-2.48 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.08 (1H, dd, J=10.1, 13.8 Hz), 3.17 (1H, dd, J=5.3, 13.8 Hz), 3.64 (3H, s), 4.66 (1H, ddd, J=5.3, 7.8, 10.1 Hz), 7.16-7.23 (1H, m), 7.24-7.32 (4H, m), 7.43 (1H, t, J=7.7 Hz), 7.54 (1H, td, J=1.3, 7.7 Hz), 7.73 (1H, td, J=1.4, 7.8 Hz), 7.83 (1H, t, J=1.5 Hz), 8.93 (1H, d, J=7.8 Hz) ppm.

(128) (.sup.13C) DMSO-d.sub.6: 18.22, 23.91, 31.18, 34.78, 36.14, 36.62, 51.97, 54.25, 80.16, 91.17, 123.21, 126.48, 127.01, 128.23, 128.76, 129.00, 129.94, 133.87, 134.05, 137.64, 165.60, 171.30, 172.05 ppm.

(129) MS(ES+) m/z 421 (M+H).

(S,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoate VSN 55

(130) ##STR00046##

(131) Following the general procedure for the Lindlar reduction, the hydrogenation of (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate (0.5 g, 1.19 mmol) gave the named product along with the trans double bond isomer (5%) and fully saturated product (10%) (determined by .sup.1H NMR). Separation by column chromatography (1-2% MeOH in DCM) gave the title compound (0.33 g, 64.4% yield). The other 2 components were not isolated.

(132) (.sup.1H) DMSO-d.sub.6: 1.64 (2H, qn, J=7.4 Hz), 2.23-2.36 (4H, m), 2.77 (3H, s), 2.90 (3H, s), 3.09 (1H, dd, J=10.3, 13.7 Hz), 3.17 (1H, dd, J=5.1, 13.7 Hz), 3.64 (3H, s), 4.66 (1H, ddd, J=5.2, 7.9, 10.2 Hz), 5.74 (1H, dt, J=7.3, 11.8 Hz), 6.45 (1H, br d, J=11.7 Hz), 7.13-7.23 (1H, m), 7.24-7.33 (4H, m), 7.40-7.47 (2H, m), 7.61-7.69 (2H, m), 8.87 (1H, d, J=7.9 Hz) ppm.

(133) (.sup.13C) DMSO-d.sub.6: 24.76, 27.68, 31.70, 34.73, 36.17, 36.60, 51.94, 54.27, 125.64, 126.46, 127.35, 128.21, 128.28, 129.05, 131.44, 133.33, 133.75, 137.09, 137.71, 166.40, 171.59, 172.15 ppm.

(134) MS(ES+) m/z 423 (M+H).

(S,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoic acid VSN 56

(135) ##STR00047##

(136) Following the general procedure for saponification, the reaction of (S,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoate (0.20 g, 0.47 mmol) with lithium hydroxide (23 mg, 0.95 mmol) gave (S,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoic acid (131 mg, 66.4% yield) as a colourless gum.

(137) (.sup.1H) DMSO-d.sub.6: 1.64 (2H, qn, J=7.4 Hz), 2.24-2.35 (4H, m), 2.77 (3H, s), 2.90 (3H, s), 3.06 (1H, dd, J=10.8, 13.7 Hz), 3.19 (1H, dd, J=4.3, 13.7 Hz), 5.73 (1H, td, J=7.3, 11.7 Hz), 6.45 (1H, d, J=11.8 Hz), 7.14-7.21 (2H, m), 7.24-7.28 (2H, m), 7.29-7.35 (2H, m), 7.42 (2H, d, J=5.3 Hz), 7.61-7.70 (2H, m), 8.72 (1H, d, J=8.2 Hz), 12.76 (1H, s) ppm.

(138) (.sup.13C) DMSO-d.sub.6: 24.78, 27.69, 31.71, 34.75, 36.21, 36.61, 54.20, 125.30, 126.32, 127.33, 128.15, 128.24, 128.31, 129.03, 131.31, 133.28, 134.01, 137.05, 138.19, 166.32, 171.60, 173.14 ppm.

(139) MS(ES+) m/z 409 (M+H).

(140) Synthesis of VSN 57 to 59

(R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate VSN 57

(141) ##STR00048##

(142) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.70 g, 2.65 mmol), (R)-methyl 2-amino-3-hydroxypropanoate.HCl (0.45 g, 2.91 mmol), DIPEA (1.4 ml, 7.94 mmol) and HATU (1.21 g, 3.17 mmol) in dry DCM (10 mL) after purification by chromatography (1-5% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate (0.77 g, 79% yield) as a viscous pale yellow oil.

(143) (.sup.1H) DMSO-d.sub.6: 1.78 (2H, qn, J=7.2 Hz), 2.42-2.49 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.65 (3H, s), 3.79 (2H, t, J=5.9 Hz), 4.52 (1H, dt, J=5.5, 7.4 Hz), 5.04 (1H, t, J=6.2 Hz), 7.47 (1H, t, J=7.7 Hz), 7.57 (1H, td, J=1.3, 7.7 Hz), 7.83 (1H, td, J=1.5, 7.8 Hz), 7.93 (1H, t, J=1.5 Hz), 8.68 (1H, d, J=7.4 Hz) ppm.

(144) (.sup.13C) DMSO-d.sub.6: 18.24, 23.93, 31.20, 34.78, 36.63, 51.88, 55.71, 60.94, 80.18, 91.16, 123.21, 127.12, 128.76, 130.07, 133.99, 134.04, 165.73, 170.92, 171.30 ppm.

(145) MS(ES+) m/z 361 (M+H).

(R,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoate VSN 58

(146) ##STR00049##

(147) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-hydroxypropanoate (0.50 g, 1.39 mmol) gave the named product along with the trans double bond isomer (5%) and fully saturated product (5%) (determined by .sup.1H NMR). Separation by column chromatography (1-3% MeOH in DCM) gave the title compound (0.32 g, 62.4% yield). The other 2 components were not isolated.

(148) (.sup.1H) DMSO-d.sub.6: 1.66 (2H, qn, J=7.4 Hz), 2.20-2.39 (4H, m), 2.78 (3H, s), 2.92 (3H, s), 3.66 (3H, s), 3.80 (2H, t, J=5.8 Hz), 4.54 (1H, dt, J=5.4, 7.4 Hz), 5.05 (1H, t, J=6.2 Hz), 5.76 (1H, dt, J=7.3, 11.7 Hz), 6.49 (1H, br d, J=11.7 Hz), 7.42-7.53 (2H, m), 7.71-7.82 (2H, m), 8.60 (1H, d, J=7.5 Hz) ppm.

(149) (.sup.13C) DMSO-d.sub.6: 24.78, 27.71, 31.72, 34.74, 36.62, 51.86, 55.66, 60.99, 125.66, 127.56, 128.28, 128.33, 131.36, 133.33, 133.81, 137.15, 166.44, 171.02, 171.61 ppm.

(150) MS(ES+) m/z 363 (M+H).

(R,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoic acid VSN 59

(151) ##STR00050##

(152) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoate (0.15 g, 0.41 mmol) with lithium hydroxide (25 mg, 1.04 mmol) gave (R,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-hydroxypropanoic acid (77 mg, 52.3% yield) as a colourless gum.

(153) (.sup.1H) DMSO-d.sub.6: 1.65 (2H, qn, J=7.4 Hz), 2.25-2.38 (4H, m), 2.77 (3H, s), 2.91 (3H, s), 3.79 (2H, d, J=5.2 Hz), 4.47 (1H, dt, J=5.2, 7.6 Hz), 5.60-5.89 (1H, m), 6.49 (1H, d, J=11.7 Hz), 7.34-7.59 (3H, m), 7.64-7.88 (2H, m), 8.41 (1H, d, J=7.7 Hz), 12.58 (1H, s) ppm.

(154) (.sup.13C) DMSO-d.sub.6: 24.79, 27.72, 31.73, 34.75, 36.63, 55.65, 61.16, 125.61, 127.53, 128.26, 128.37, 131.25, 133.29, 134.07, 137.13, 166.32, 171.61, 171.90 ppm.

(155) MS(ES+) m/z 349 (M+H).

(156) Synthesis of VSN 60 to 62

(R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate VSN 60

(157) ##STR00051##

(158) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.70 g, 2.65 mmol), (R)-methyl 2-amino-3-phenylpropanoate.HCl (0.6 g, 2.78 mmol), DIPEA (1.4 ml, 7.94 mmol) and HATU (1.3 g, 3.44 mmol) in dry DCM (10 mL) after purification by chromatography (1-4% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate (0.81 g, 68.4% yield) as a viscous pale yellow oil.

(159) (.sup.1H) DMSO-d.sub.6: 1.74-1.82 (2H, m), 2.42-2.48 (4H, m), 2.82 (3H, s), 2.97 (3H, s), 3.08 (1H, dd, J=10.1, 13.8 Hz), 3.17 (1H, dd, J=5.3, 13.8 Hz), 3.64 (3H, s), 4.66 (1H, ddd, J=5.3, 7.8, 10.1 Hz), 7.16-7.23 (1H, m), 7.24-7.32 (4H, m), 7.43 (1H, t, J=7.7 Hz), 7.54 (1H, td, J=1.3, 7.7 Hz), 7.73 (1H, td, J=1.4, 7.8 Hz), 7.83 (1H, t, J=1.5 Hz), 8.93 (1H, d, J=7.8 Hz) ppm.

(160) (.sup.13C) DMSO-d.sub.6: 18.22, 23.91, 31.18, 34.78, 36.14, 36.62, 51.96, 54.25, 80.16, 91.17, 123.21, 126.48, 127.01, 128.22, 128.75, 129.00, 129.94, 133.87, 134.04, 137.63, 165.60, 171.29, 172.05 ppm.

(161) MS(ES+) m/z 421 (M+H).

(R,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoate VSN 61

(162) ##STR00052##

(163) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)-3-phenylpropanoate (0.50 g, 1.19 mmol) gave the named product along with the trans double bond isomer (5%) and fully saturated product (10%) (determined by .sup.1H NMR). Separation by column chromatography (1-2% MeOH in DCM) gave the title compound (0.37 g, 72.2% yield). The other 2 components were not isolated.

(164) (.sup.1H) DMSO-d.sub.6: 1.65 (2H, qn, J=7.4 Hz), 2.25-2.38 (4H, m), 2.78 (3H, s), 2.91 (3H, s), 3.10 (1H, dd, J=10.3, 13.7 Hz), 3.18 (1H, dd, J=5.1, 13.7 Hz), 3.65 (3H, s), 4.67 (1H, ddd, J=5.2, 7.9, 10.2 Hz), 5.75 (1H, dt, J=7.4, 11.8 Hz), 6.46 (1H, br d, J=11.6 Hz), 7.15-7.24 (1H, m), 7.24-7.36 (4H, m), 7.44 (2H, d, J=5.0 Hz), 7.61-7.71 (2H, m), 8.87 (1H, d, J=8.0 Hz) ppm.

(165) (.sup.13C) DMSO-d.sub.6: 24.76, 27.68, 31.70, 34.73, 36.17, 36.60, 51.94, 54.27, 125.64, 126.46, 127.35, 128.21, 128.28, 129.04, 131.43, 133.33, 133.75, 137.09, 137.71, 166.40, 171.59, 172.15 ppm.

(166) MS(ES+) m/z 423 (M+H).

(R,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoic acid VSN 62

(167) ##STR00053##

(168) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoate (0.20 g, 0.47 mmol) with lithium hydroxide (23 mg, 0.95 mmol) gave (R,Z)-2-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)-3-phenylpropanoic acid (0.18 g, 0.43 mmol, 91% yield) as a colourless gum.

(169) (.sup.1H) DMSO-d.sub.6: 1.66 (2H, qn, J=7.4 Hz), 2.24-2.35 (4H, m), 2.77 (3H, s), 2.91 (3H, s), 3.08 (1H, dd, J=10.5, 13.8 Hz), 3.16-3.22 (1H, m), 4.64 (1H, td, J=4.5, 10.2 Hz), 5.74 (1H, dt, J=7.3, 11.7H), 6.45 (1H, d, J=11.7 Hz), 7.18 (1H, t, J=7.1 Hz), 7.22-7.34 (4H, m), 7.42 (2H, d, J=5.1 Hz), 7.59-7.71 (2H, m), 8.62 (1H, br d, J=4.4 Hz), 12.63 (1H, s) ppm.

(170) (.sup.13C) DMSO-d.sub.6: 24.78, 27.69, 31.71, 34.75, 36.21, 36.61, 54.21, 125.60, 126.32, 127.33, 128.15, 128.24, 128.32, 129.04, 131.31, 133.28, 134.01, 137.05, 138.20, 166.31, 171.60, 173.15 ppm.

(171) MS(ES+) m/z 409 (M+H).

(172) Synthesis of VSN 63 to 65

(S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate VSN 63

(173) ##STR00054##

(174) To a solution of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (1.50 g, 4.63 mmol), (S)-methyl 2-aminopropanoate.HCl (0.78 g, 5.55 mmol) and HATU (2.3 g, 6.02 mmol) in dry DMF (15 mL) was added DIPEA (2.5 ml, 13.88 mmol). The reaciton was stirred at it until complete by LC-MS. Next, the reaction mixture was poured into water (150 mL) and extracted with EtOAc (4100 mL) before the combined organic extracts were washed with sat. aq. ammonium chloride (100 mL), water (550 mL) then brine (50 mL) and dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude material was purified by chromatography (25-100% EtOAc in iso-hexanes) to give (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (1.17 g, 68.3% yield at 90% purity). Material of sufficient purity to proceed. A sample (100 mg) was purified by preparative HPLC (20-50% MeCN in water (0.1% formic)) to give analytically pure material.

(175) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.3 Hz), 1.77 (2H, qn, J=7.2 Hz), 2.43-2.48 (4H, m), 4.82 (3H, s), 2.97 (3H, s), 3.64 (3H, s), 4.47 (1H, qn, J=7.32 Hz), 7.46 (1H, t, J=7.7 Hz), 7.56 (1H, td, J=1.3, 7.7 Hz), 7.82 (1H, td, J=1.5, 7.8 Hz), 7.91-7.94 (1H, m), 8.89 (1H, d, J=6.9 Hz) ppm.

(176) (.sup.13C) DMSO-d.sub.6: 16.68, 18.24, 23.94, 31.21, 34.80, 36.64, 48.31, 51.93, 80.20, 91.15, 123.22, 127.13, 128.78, 130.05, 133.90, 134.03, 165.43, 171.31 ppm.

(177) MS(ES+) m/z 345 (M+H).

(S)-methyl 2-(3-(6-(dimethylamino)-6-oxohexyl)benzamido)propanoate VSN 64

(178) ##STR00055##

(179) Following the general procedure for the reduction of an alkyne to alkane, the hydrogenation of (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (0.37 g, 1.07 mmol) after purification by preparative HPLC (20-50% MeCN in water (0.1% formic)) gave the title compound (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohexyl) benzamido)propanoate (228 mg, 59.7% yield) as a colourless viscous oil.

(180) (.sup.1H) DMSO-d.sub.6: 1.26-1.35 (2H, m), 1.40 (3H, t, J=7.4 Hz), 1.48-1.55 (2H, m), 1.56-1.64 (2H, m), 2.26 (2H, t, J=7.4 Hz), 2.60-2.64 (2H, m), 2.79 (3H, s), 2.93 (3H, s), 3.64 (3H, s), 4.47 (1H, qn, J=7.2 Hz), 7.35-7.39 (2H, m), 7.66-7.71 (2H, m), 8.74 (1H, d, J=7.0 Hz) ppm.

(181) (.sup.13C) DMSO-d.sub.6: 16.73, 24.48, 28.44, 30.82, 32.24, 34.73, 34.97, 36.67, 48.22, 51.85, 124.90, 127.26, 128.16, 131.42, 133.62, 142.41, 166.31, 171.84, 173.20 ppm.

(182) MS(ES+) m/z 349 (M+H).

(S)-2-(3-(6-(dimethylamino)-6-oxohexyl)benzamido)propanoic acid VSN 65

(183) ##STR00056##

(184) Following the general procedure for saponification, the reaction of (S)-methyl 2-(3-(6-(dimethylamino)-6-oxohexyl)benzamido)propanoate (0.10 g, 0.287 mmol) and lithium hydroxide (14 mg, 0.57 mmol) gave (S)-2-(3-(6-(dimethylamino)-6-oxohexyl) benzamido)propanoic acid (78 mg, 81% yield) as a white solid.

(185) (.sup.1H) DMSO-d.sub.6: 1.25-1.35 (2H, m), 1.39 (3H, d, J=7.4 Hz), 1.46-1.56 (2H, m), 1.56-1.65 (2H, m), 2.26 (2H, t, J=7.4 Hz), 2.62 (2H, t, J=7.7 Hz), 2.79 (3H, s), 2.93 (3H, s), 4.41 (1H, qn, J=7.3 Hz), 7.36 (2H, dd, J=1.2, 4.0 Hz), 7.63-7.76 (2H, m), 8.60 (1H, d, J=7.2 Hz), 12.50 (1H, s) ppm.

(186) (.sup.13C) DMSO-d.sub.6: 16.89, 24.48, 28.44, 30.83, 32.25, 34.74, 34.99, 36.67, 48.08, 124.88, 127.24, 128.11, 131.29, 133.89, 142.36, 166.20, 171.85, 174.23 ppm.

(187) MS(ES+) m/z 335 (M+H).

(188) Synthesis of VSN 66-68

N,N-dimethylpent-4-ynamide

(189) ##STR00057##

(190) To a solution of pent-4-ynoic acid (3.1 g, 31.6 mmol) in dry DCM (30 mL) and DMF (1 drop) at 0 C. was added oxalyl chloride (4.01 ml, 47.4 mmol). The reaction was allowed to warm to it and stir for 1 h before the volatiles were removed in vacuo. The residue was redissolved in dry THF (10 mL) and added drop-wise to a cooled (ice bath) solution of dimethylamine (40 wt % in water) (20.0 ml, 158 mmol). The reaction was stirred in the ice bath for 1 h then extracted with DCM (330 mL). The combined organic extracts were washed with water (50 mL) and dried (MgSO.sub.4), filtered then concentrated in vacuo to give N,N-dimethylpent-4-ynamide (3.3 g, 79% yield) as a brown free-flowing oil that solidified on standing.

(191) (.sup.1H) DMSO-d.sub.6: 2.31-1.35 (2H, m), 2.49-2.51 (2H, m), 2.74 (1H, t, J=2.6 Hz), 2.81 (3H, s), 2.94 (3H, s) ppm.

(R)-methyl 2-(3-(5-(dimethylamino)-5-oxopent-1-yn-1-yl)benzamido)propanoate VSN 66

(192) ##STR00058##

(193) Following the general method for Sonogashira coupling, the reaction of (R)-methyl 2-(3-iodobenzamido)propanoate (2.0 g, 4.80 mmol) and N,N-dimethylpent-4-ynamide (0.73 g, 5.52 mmol) after purification by column chromatography (1-3% MeOH in DCM) gave (R)-methyl 2-(3-(5-(dimethylamino)-5-oxopent-1-yn-1-yl)benzamido)propanoate (1.4 g, 86% yield).

(194) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 2.64 (4H, br s), 2.82 (3H, s), 2.99 (3H, s), 3.65 (3H, s), 4.48 (1H, qn, J=7.2 Hz), 7.46 (1H, t, J=7.8 Hz), 7.55 (1H, td, J=1.4, 7.7 Hz), 7.83 (1H, td, J=1.6, 7.9 Hz), 7.92 (1H, t, J=1.4 Hz), 8.89 (1H, d, J=6.9 Hz) ppm.

(195) (.sup.13C) DMSO-d.sub.6: 14.77, 16.66, 31.59, 34.89, 36.53, 48.28, 51.89, 79.63, 91.22, 123.19, 127.09, 128.74, 130.03, 133.92, 133.95, 165.42, 170.16, 173.05 ppm.

(196) MS(ES+) m/z 331 (M+H).

(R)-methyl 2-(3-(5-(dimethylamino)-5-oxopentyl)benzamido)propanoate VSN 67

(197) ##STR00059##

(198) Following the general procedure for the reduction of an alkyne to alkane, the hydrogenation of (R)-methyl 2-(3-(5-(dimethylamino)-5-oxopent-1-yn-1-yl)benzamido)propanoate (400 mg, 1.211 mmol) after purification by chromatography (1-3% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(5-(dimethylamino)-5-oxopentyl)benzamido)propanoate (0.36 g, 87% yield) as a colourless oil.

(199) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.45-1.56 (2H, m), 1.56-1.69 (2H, m), 2.30 (2H, t, J=7.3 Hz), 2.64 (2H, t, J=7.5 Hz), 2.79 (3H, s), 2.93 (3H, s), 3.64 (3H, s), 4.32-4.73 (1H, m), 7.37 (2H, dd, J=1.0, 4.1 Hz) 7.58-7.87 (2H, m), 8.74 (1H, d, J=6.9 Hz) ppm.

(200) (.sup.13C) DMSO-d.sub.6: 16.74, 24.26, 30.53, 32.11, 34.74, 34.85, 36.67, 48.23, 51.85, 124.90, 127.25, 128.16, 131.42, 133.63, 142.32, 166.32, 171.79, 173.20 ppm.

(201) MS(ES+) m/z 335 (M+H).

(R)-2-(3-(5-(dimethylamino)-5-oxopentyl)benzamido)propanoic acid VSN 68

(202) ##STR00060##

(203) Following the general saponification procedure, the reaction of (R)-methyl 2-(3-(5-(dimethylamino)-5-oxopentyl)benzamido)propanoate (0.15 g, 0.45 mmol) with lithium hydroxide (16 mg, 0.67 mmol) gave (R)-2-(3-(5-(dimethylamino)-5-oxopentyl)benzamido)propanoic acid (82 mg, 55.9% yield) as a waxy white solid.

(204) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.4 Hz), 1.46-1.54 (2H, m), 1.58-1.65 (2H, m), 2.30 (2H, t, J=7.3 Hz), 2.64 (2H, t, J=7.5 Hz), 2.79 (3H, s), 2.93 (3H, s), 4.41 (1H, qn, J=7.3 Hz), 7.34-7.39 (2H, m), 7.65-7.74 (2H, m), 8.61 (1H, d, J=7.2 Hz), 12.54 (1H, br s) ppm.

(205) (.sup.13C) DMSO-d.sub.6: 16.91, 24.27, 30.55, 32.12, 34.75, 34.86, 36.68, 48.10, 124.88, 127.22, 128.12, 131.29, 133.91, 142.26, 166.20, 171.80, 174.23 ppm.

(206) MS(ES+) m/z 321 (M+H).

(207) Synthesis of VSN 69 to 71

(R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate VSN 69

(208) ##STR00061##

(209) Using the general procedure described for amide coupling, the reaction of (R)-6-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)hex-5-ynoic acid (2.41 g, 5.70 mmol), dimethylamine.HCl (0.56 g, 6.84 mmol), HATU (2.60 g, 6.84 mmol) and TEA (1.99 ml, 14.24 mmol) in dry DCM (30 mL) after purification by chromatography (25-100% EtOAc in iso-hexanes) gave the title compound (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (1.94 g, 89% yield) as a colourless oil. A sample (150 mg) was purified by preparative HPLC (20-50% MeCN in water (0.1% formic)) to give analytically pure material (128 mg).

(210) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.78 (2H, qn, J=7.1 Hz), 2.43-2.50 (4H, m), 2.83 (3H, s), 2.98 (3H, s), 3.65 (3H, s), 4.48 (1H, qn, J=7.3 Hz), 7.47 (1H, t, J=7.8 Hz), 7.57 (1H, td, J=1.3, 7.6 Hz), 7.83 (1H, td, J=1.4, 7.8 Hz), 7.93 (1H, t, J=1.5 Hz), 8.90 (1H, d, J=7.0 Hz) ppm.

(211) (.sup.13C) DMSO-d.sub.6: 16.69, 18.25, 23.95, 31.22, 34.81, 36.65, 48.31, 51.94, 91.16, 123.22, 127.13, 128.79, 130.06, 133.91, 134.05, 165.44, 171.32, 173.10 ppm.

(212) MS(ES+) m/z 345 (M+H).

(R)-methyl 2-(3-(6-(dimethylamino)-6-oxohexyl)benzamido)propanoate VSN 70

(213) ##STR00062##

(214) Following the general procedure for the reduction of an alkyne to alkane, the hydrogenation of (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (250 mg, 0.73 mmol) after purification by preparative HPLC (20-50% MeCN in water (0.2% formic)) gave the title compound (R)-methyl 2-(3-(6-(dimethylamino)-6-oxohexyl) benzamido)propanoate (172 mg, 66.6% yield) as a colourless viscous oil.

(215) (.sup.1H) DMSO-d.sub.6: 1.21-1.36 (2H, m), 1.40 (3H, d, J=7.3 Hz), 1.51 (2H, qn, J=7.4 Hz), 1.60 (2H, qn, J=7.6 Hz), 2.26 (2H, t, J=7.4 Hz), 2.57-2.68 (2H, m), 2.79 (3H, s), 2.93 (3H, s), 3.64 (3H, s), 4.47 (1H, qn, J=7.3 Hz), 7.37 (2H, d, J=5.0 Hz), 7.65-7.73 (2H, m), 8.74 (1H, d, J=6.9 Hz) ppm.

(216) (.sup.13C) DMSO-d.sub.6: 16.73, 24.48, 28.44, 30.82, 32.24, 34.73, 34.98, 36.67, 48.22, 51.85, 124.90, 127.26, 128.16, 131.42, 133.62, 142.42, 166.31, 171.85, 173.20 ppm.

(217) MS(ES+) m/z 349 (M+H).

(R)-2-(3-(6-(dimethylamino)-6-oxohexyl)benzamido)propanoic acid VSN 71

(218) ##STR00063##

(219) Following the general procedure for the reduction of an alkyne to alkane, the hydrogenation of (R)-2-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoic acid (146 mg, 0.44 mmol) gave the title compound (R)-2-(3-(6-(dimethylamino)-6-oxohexyl) benzamido)propanoic acid (0.14 g, 93% yield) as a colourless viscous oil. No purification required.

(220) (.sup.1H) DMSO-d.sub.6: 1.26-1.36 (2H, m), 1.40 (3H, d, J=7.4 Hz), 1.48-1.57 (2H, m), 1.57-1.66 (2H, m), 2.27 (2H, t, J=7.4 Hz), 2.59-2.67 (2H, m), 2.79 (3H, s), 2.94 (3H, s), 4.41 (1H, qn, J=7.4 Hz), 7.32-7.42 (2H, m), 7.65-7.76 (2H, m), 8.60 (1H, d, J=7.2 Hz), 12.51 (1H, s) ppm.

(221) (.sup.13C) DMSO-d.sub.6: 16.93, 24.53, 28.49, 30.90, 32.28, 34.77, 35.04, 36.70, 48.14, 54.95, 124.92, 127.28, 128.16, 131.35, 133.90, 142.40, 166.23, 171.89, 174.31 ppm.

(222) MS(ES+) m/z 335 (M+H).

(223) Synthesis of VSN 72 to 74

oct-7-ynoic acid

(224) ##STR00064##

(225) To a solution of 6-bromohexanoic acid (2.4 g, 12.30 mmol) in dry DMSO (20 mL) under nitrogen was was added lithium acetylide ethylenediamine complex (2.49 g, 27.1 mmol) protion-wise over 15 min. Upon complete addition, the resulting brown solution was stirred at it for 2 h. The reaction was then quenched by the addition of water (20 mL) and acidified to pH 1 with 1 N HCl. The product was then extracted with EtOAc (460 mL) before the combined organic extracts were washed with water (580 mL) and brine (60 mL) then dried (MgSO4), filtered and concentrated in vacuo to give oct-7-ynoic acid (0.7 g, 36.5% yield) as a pale orange oil.

(226) (.sup.1H) DMSO-d.sub.6: 1.30-1.38 (2H, m), 1.39-1.46 (2H, m), 1.39-1.54 (2H, m), 2.14 (2H, dt, J=2.7, 7.0 Hz), 2.20 (2H, t, J=7.4 Hz), 2.75 (1H, t, J=2.7 Hz), 11.98 (1H, s) ppm.

(R)-8-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)oct-7-ynoic acid

(227) ##STR00065##

(228) Following the general method for Sonogashira coupling, the reaction of (R)-methyl 2-(3-iodobenzamido)propanoate (1.8 g, 4.34 mmol) and oct-7-ynoic acid (0.7 g, 4.99 mmol) after purification by column chromatography (1-3% MeOH in DCM) gave (R)-8-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)oct-7-ynoic acid (1.3 g, 69.3% yield @ 80% purity).

(229) (.sup.1H) DMSO-d.sub.6: 1.35-1.49 (5H, m), 1.50-1.63 (4H, m), 2.23 (2H, t, J=7.2 Hz), 2.44 (2H, t, J=7.0 Hz), 3.64 (3H, s), 4.47 (1H, qn, J=7.3, 1.8 Hz), 7.44-7.50 (1H, m), 7.55 (1H, td, J=1.4, 7.7 Hz), 7.82 (1H, dt, J=1.5, 7.8 Hz), 7.86-7.95 (1H, m), 8.83-8.90 (1H, m), 12.00 (1H, s) ppm.

(230) MS(ES+) m/z 346 (M+H).

(R)-methyl 2-(3-(8-(dimethylamino)-8-oxooct-1-yn-1-yl)benzamido)propanoate VSN 72

(231) ##STR00066##

(232) Using the general procedure described for amide coupling, the reaction of (R)-8-(3-((1-methoxy-1-oxopropan-2-yl)carbamoyl)phenyl)oct-7-ynoic acid (1.3 g, 3.01 mmol), dimethylamine.HCl (0.27 g, 3.31 mmol), DIPEA (1.32 ml, 7.53 mmol) and HATU (1.32 g, 3.46 mmol) in dry DCM (15 mL) after purification by chromatography (1-3% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(8-(dimethylamino)-8-oxooct-1-yn-1-yl)benzamido)propanoate (1.0 g, 85% yield) as a viscous yellow oil.

(233) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.3 Hz), 1.42-1.47 (2H, m), 1.48-1.61 (4H, m), 2.30 (2H, t, J=7.5 Hz), 2.44 (2H, t, J=7.0 Hz), 2.79 (3H, s), 2.95 (3H, s), 3.64 (3H, s), 4.47 (1H, qn, J=7.2 Hz), 7.45 (1H, t, J=7.8 Hz), 7.55 (1H, td, J=1.4, 7.8 Hz), 7.81 (1H, td, J=1.6, 7.9 Hz), 7.91 (1H, t, J=1.5 Hz), 8.87 (1H, d, J=6.9 Hz) ppm.

(234) (.sup.13C) DMSO-d.sub.6: 16.70, 18.52, 24.18, 27.95, 28.13, 32.19, 34.73, 36.67, 48.27, 51.87, 79.93, 91.45, 123.29, 127.02, 128.71, 129.96, 133.89, 133.97, 165.41, 171.81, 173.04 ppm.

(235) MS(ES+) m/z 373 (M+H).

(R)-methyl 2-(3-(8-(dimethylamino)-8-oxooctyl)benzamido)propanoate VSN 73

(236) ##STR00067##

(237) Following the general procedure for the reduction of an alkyne to alkane, the hydrogenation of (R)-methyl 2-(3-(8-(dimethylamino)-8-oxooct-1-yn-1-yl)benzamido)propanoate (400 mg, 1.07 mmol) after purification by chromatography (1-5% MeOH in DCM) gave the title compound (R)-methyl 2-(3-(8-(dimethylamino)-8-oxooctyl)benzamido)propanoate (0.34 g, 80% yield) as a colourless oil.

(238) (.sup.1H) DMSO-d.sub.6: 1.24-1.29 (6H, m), 1.40 (3H, d, J=7.32 Hz), 1.46 (2H, qn, J=6.9 Hz), 1.55-1.62 (2H, m), 2.24 (2H, t, J=7.5 Hz), 2.60-2.64 (2H, m), 2.79 (3H, s), 2.93 (3H, s), 3.64 (3H, s), 4.47 (1H, qn, J=7.2 Hz), 7.35-7.39 (2H, m), 7.66-7.72 (2H, m), 8.74 (1H, d, J=7.0 Hz) ppm.

(239) (.sup.13C) DMSO-d.sub.6: 6.73, 24.62, 28.58, 28.69, 28.73, 30.89, 32.28, 34.72, 35.02, 36.67, 48.22, 51.84, 124.87, 127.23, 128.14, 131.39, 133.62, 142.48, 166.31, 171.89, 173.20 ppm.

(240) MS(ES+) m/z 377 (M+H).

(R)-2-(3-(8-(dimethylamino)-8-oxooctyl)benzamido)propanoic acid VSN 74

(241) ##STR00068##

(242) Following the general procedure for saponification, the reaction of (R)-methyl 2-(3-(8-(dimethylamino)-8-oxooctyl)benzamido)propanoate (0.15 g, 0.40 mmol) with lithium hydroxide (14 mg, 0.60 mmol) gave (R)-2-(3-(8-(dimethylamino)-8-oxooctyl) benzamido)propanoic acid (0.11 g, 74.6% yield).

(243) (.sup.1H) DMSO-d.sub.6: 1.22-1.34 (6H, m), 1.40 (3H, d, J=7.4 Hz), 1.47 (2H, qn, J=7.4 Hz), 1.56-1.63 (2H, m), 2.25 (2H, t, J=7.4 Hz), 2.61-2.65 (2H, m), 2.79 (3H, s), 2.94 (3H, s), 4.42 (1H, qn, J=7.3 Hz), 7.35-7.39 (2H, m), 7.68-7.72 (2H, m) 8.61 (1H, d, J=7.3 Hz), 12.52 (1H, s) ppm.

(244) (.sup.13C) DMSO-d.sub.6: 6.89, 24.63, 28.60, 28.70, 28.74, 30.91, 32.29, 34.73, 35.04, 36.68, 48.08, 124.86, 127.21, 128.10, 131.26, 133.89, 142.42, 166.21, 171.90, 174.24 ppm.

(245) MS(ES+) m/z 363 (M+H).

(246) Synthesis of VSN 75 to 77

methyl 3-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate VSN 75

(247) ##STR00069##

(248) Using the general procedure described for amide coupling, the reaction of 3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzoic acid (0.75 g, 2.314 mmol), methyl 3-aminopropanoate.HCl (0.36 g, 2.55 mmol), DIPEA (1.24 ml, 6.94 mmol) and HATU (1.14 g, 3.01 mmol) in dry DCM (15 mL) after purification by chromatography (1-3% MeOH in DCM) gave the title compound methyl 3-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (0.66 g, 79% yield) as a viscous pale yellow oil.

(249) (.sup.1H) DMSO-d.sub.6: 1.78 (2H, qn, J=7.2 Hz), 2.41-2.48 (4H, m), 2.59 (2H, t, J=7.0 Hz), 2.82 (3H, s), 2.97 (3H, s), 3.43-3.52 (2H, m), 3.60 (3H, s), 7.44 (1H, t, J=7.8 Hz), 7.53 (1H, td, J=1.3, 7.7 Hz), 7.77 (1H, td, J=1.4, 7.8 Hz), 7.84 (1H, t, J=1.5 Hz), 8.61 (1H, t, J=5.4 Hz) ppm.

(250) (.sup.13C) DMSO-d.sub.6: 18.22, 23.93, 31.19, 33.41, 34.77, 35.51, 36.62, 51.38, 80.23, 91.04, 123.17, 126.81, 128.70, 129.77, 133.72, 134.55, 165.45, 171.29, 171.70 ppm.

(251) MS(ES+) m/z 345 (M+H).

(Z)-methyl 3-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)propanoate VSN 76

(252) ##STR00070##

(253) Following the general procedure for the Lindlar reduction, the hydrogenation of methyl 3-(3-(6-(dimethylamino)-6-oxohex-1-yn-1-yl)benzamido)propanoate (0.40 g, 1.16 mmol) gave the named product along with the trans double bond isomer (7%) and fully saturated product (25%) (determined by .sup.1H NMR). Separation by column chromatography (1-3% MeOH in DCM) gave the title compound (0.20 g, 48.7% yield). The other 2 components were not isolated.

(254) (.sup.1H) DMSO-d.sub.6: 1.64 (2H, qn, J=7.4 Hz), 2.25-2.34 (4H, m), 2.60 (2H, t, J=7.0 Hz), 2.78 (3H, s), 2.92 (3H, s), 3.45-3.54 (2H, m), 3.60 (3H, s), 5.73 (1H, td, J=7.3, 11.7 Hz), 6.46 (1H, br d, J=11.7 Hz), 7.43 (2H, dd, J=1.3, 3.9 Hz), 7.64-7.75 (2H, m), 8.57 (1H, t, J=5.4 Hz) ppm.

(255) (.sup.13C) DMSO-d.sub.6: 24.75, 27.72, 31.71, 33.54, 34.74, 35.51, 36.61, 51.36, 125.42, 127.16, 128.25, 128.36, 131.11, 133.22, 134.39, 137.08, 166.22, 171.63, 171.73 ppm.

(256) MS(ES+) m/z 347 (M+H).

(Z)-3-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)propanoic acid VSN 77

(257) ##STR00071##

(258) Following the general procedure for saponification, the reaction of (Z)-methyl 3-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)propanoate (0.1 g, 0.29 mmol) with lithium hydroxide (14 mg, 0.58 mmol) gave (Z)-3-(3-(6-(dimethylamino)-6-oxohex-1-en-1-yl)benzamido)propanoic acid (86 mg, 88% yield) as a colourless gum.

(259) (.sup.1H) DMSO-d.sub.6: 1.64 (2H, qn, J=7.4 Hz), 2.27-2.32 (4H, m), 2.50-2.55 (2H, m), 2.78 (3H, s), 2.92 (3H, s), 3.46 (2H, q, J=7.1 Hz), 5.73 (1H, dt, J=7.3, 11.7 Hz), 6.46 (1H, d, J=11.7 Hz), 7.43 (2H, dd, J=3.9, 1.4 Hz), 7.67-7.71 (1H, m), 7.72 (1H, br s), 8.55 (1H, t, J=5.5 Hz), 12.20 (1H, s) ppm.

(260) (.sup.13C) DMSO-d.sub.6: 24.76, 27.73, 31.72, 33.76, 34.75, 35.59, 36.63, 125.41, 127.18, 128.24, 128.38, 131.07, 133.21, 134.45, 137.08, 166.14, 171.66, 172.86 ppm.

(261) MS(ES+) m/z 333 (M+H).

(262) Synthesis of VSN 78 to 80

(R)-methyl 2-(3-(5-cyanopent-1-yn-1-yl)benzamido)propanoate VSN 78

(263) ##STR00072##

(264) Following the general method for Sonogashira coupling, the reaction of (R)-methyl 2-(3-iodobenzamido)propanoate (1.6 g, 4.56 mmol) and hex-5-ynenitrile (0.72 ml, 6.84 mmol) after purification by chromatography (20-40% EtOAc in iso-hexanes) gave (R)-methyl 2-(3-(5-cyanopent-1-yn-1-yl)benzamido)propanoate (1.3 g, 93% yield).

(265) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.3 Hz), 1.86 (2H, qn, J=7.1 Hz), 2.57 (2H, t, J=7.0 Hz), 2.65 (2H, t, J=7.2 Hz), 3.64 (3H, s), 4.47 (1H, qn, J=7.2 Hz), 7.47 (1H, t, J=7.8 Hz), 7.58-7.60 (1H, m), 7.82-7.85 (1H, m), 7.94 (1H, t, J=1.4 Hz), 8.87 (1H, d, J=6.9 Hz) ppm.

(266) (.sup.13C) DMSO-d.sub.6: 15.58, 16.66, 17.86, 24.06, 48.29, 51.90, 80.88, 89.35, 120.18, 122.90, 127.27, 128.73, 130.11, 133.93, 134.10, 165.40, 173.05 ppm.

(267) MS(ES+) m/z 299.2 (M+H).

(R,Z)-methyl 2-(3-(5-cyanopent-1-en-1-yl)benzamido)propanoate VSN 79

(268) ##STR00073##

(269) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(5-cyanopent-1-yn-1-yl)benzamido)propanoate (0.50 g, 1.68 mmol) gave the named product with trace amounts of the trans double bond isomer and fully saturated products (determined by .sup.1H NMR). Separation by column chromatography (1-2% MeOH in DCM) gave the title compound (0.42 g, 82% yield). The other 2 components were not isolated.

(270) (.sup.1H) DMSO-d.sub.6: 1.41 (3H, d, J=7.3 Hz), 1.73 (2H, qn, J=7.3 Hz), 2.40 (2H, dq, J=1.8, 7.1 Hz), 2.53-2.55 (2H, m), 3.65 (3H, s), 4.49 (1H, qn, J=7.2 Hz), 5.74 (1H, td, J=7.2, 11.6 Hz), 6.53 (1H, dt, J=1.6, 11.7 Hz), 7.47-7.49 (2H, m), 7.76-7.78 (2H, m), 8.81 (1H, d, J=6.9 Hz) ppm.

(271) (.sup.13C) DMSO-d.sub.6: 15.80, 16.72, 24.97, 27.10, 48.26, 51.87, 120.45, 125.83, 127.65, 128.30, 129.14, 131.28, 131.61, 133.81, 136.85, 166.13, 173.14 ppm.

(272) MS(ES+) m/z 301.2 (M+H).

(R,Z)-2-(3-(5-cyanopent-1-en-1-yl)benzamido)propanoic acid VSN 80

(273) ##STR00074##

(274) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(5-cyanopent-1-en-1-yl)benzamido)propanoate (0.15 g, 0.50 mmol) with lithium hydroxide (24 mg, 0.99 mmol) gave (R,Z)-2-(3-(5-cyanopent-1-en-1-yl)benzamido)propanoic acid (0.12 g, 80% yield).

(275) (.sup.1H) DMSO-d.sub.6: 1.39 (3H, d, J=7.4 Hz), 1.72 (2H, qn, J=7.6 Hz), 2.39 (2H, dq, J=1.7, 7.3 Hz), 2.52-2.54 (2H, m), 4.41 (1H, qn, 7.3 Hz), 5.73 (1H, td, J=7.1, 11.7 Hz), 6.53 (1H, br d, J=11.7 Hz), 7.46-7.47 (2H, m), 7.76-7.77 (2H, m), 8.67 (1H, d, J=7.2 Hz), 12.53 (1H, br s) ppm.

(276) (.sup.13C) DMSO-d.sub.6: 15.80, 16.87, 24.98, 27.11, 48.14, 120.46, 125.81, 127.65, 128.26, 129.19, 131.16, 131.57, 134.08, 136.81, 166.02, 174.19 ppm.

(277) MS(ES+) m/z 287.2 (M+H).

(278) Synthesis of VSN 81, 85 and 86

methyl 3-(5-hydroxypent-1-yn-1-yl)benzoate

(279) ##STR00075##

(280) Following the general method for Sonogashira coupling, the reaction of methyl 3-iodobenzoate (7.45 g, 28.4 mmol) and pent-4-yn-1-ol (3.97 ml, 42.6 mmol) after purification by chromatography (10-50% EtOAc in iso-hexanes) gave methyl 3-(5-hydroxypent-1-yn-1-yl)benzoate (5.1 g, 81% yield) as a free flowing orange oil.

(281) (.sup.1H) DMSO-d.sub.5: 1.63-1.75 (2H, m), 2.45-2.49 (2H, m), 3.46-3.57 (2H, m), 3.86 (3H, s), 4.55 (1H, t, J=5.2 Hz), 7.47-7.56 (1H, m), 7.65 (1H, td, J=1.4, 7.7 Hz), 7.86-7.93 (2H, m) ppm.

(282) MS(ES+) m/z 219 (M+H).

methyl 3-(5-(acetylthio)pent-1-yn-1-yl)benzoate

(283) ##STR00076##

(284) To a solution of methyl 3-(5-hydroxypent-1-yn-1-yl)benzoate (1.93 g, 8.84 mmol) and triethylamine (1.54 ml, 11.05 mmol) in dry DCM (20 mL) under nitrogen and cooled in an ice bath was added methanesulfonyl chloride (0.75 ml, 9.73 mmol) over 5 mins. The reaction mixture was then stirred at it until judged complete by LCMS analysis. The reaction mixture was partitioned with DCM (50 mL) and sat. aq. ammonium chloride (30 mL). The aqueous layer was extracted with DCM (215 mL) before the combined organic extracts were washed with water (20 mL) and brine (20 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo. The residue was then redissolved in dry DMF (25 mL) and treated with potassium ethanethioate (1.01 g, 8.84 mmol) and stirred at rt until judged complete by LCMS analysis. The reaction mixture was then partitioned between EtOAc (100 mL) and water (100 mL). The aqueous layer was extracted with EtOAc (330 mL) before the combined organic extracts were washed with water (530 mL), brine (30 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude material was purified by chromatography (0-10% EtOAc in iso-hexanes) to give methyl 3-(5-(acetylthio)pent-1-yn-1-yl)benzoate (2.2 g, 85% yield) as an orange free-flowing oil.

(285) (.sup.1H) DMSO-d.sub.6: 1.74-1.86 (2H, m), 2.34 (3H, s), 2.51-2.54 (2H, m), 2.95-3.03 (2H, m), 3.86 (3H, s), 7.49-7.55 (1H, m), 7.65-7.70 (1H, m), 7.89-7.94 (2H, m) ppm.

(286) MS(ES+) m/z 277 (M+H).

5-(3-(methoxycarbonyl)phenyl)pent-4-yne-1-sulfonic acid

(287) ##STR00077##

(288) Procedure followed from PCT 2003035627.

(289) To a solution of methyl 3-(5-(acetylthio)pent-1-yn-1-yl)benzoate (2.2 g, 7.96 mmol) in AcOH (10 mL) was added a solution of hydrogen peroxide (9.76 ml, 127 mmol) in AcOH (20 mL). The reaction mixture was stirred at it until complete by LC-MS analysis then cooled in an ice bath and quenched by the addition of 5% Pd/C (200 mg). The mixture was stirred for 20 min then filtered through a pad of celite and the volatiles were removed in vacuo. The residue was then azeoptroped with toluene (320 mL) to give 5-(3-(methoxycarbonyl)phenyl)pent-4-yne-1-sulfonic acid (1.9 g, 80%) as a brown semi-solid.

(290) MS(ES+) m/z 281 (M+H).

methyl 3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoate

(291) ##STR00078##

(292) A solution of 5-(3-(methoxycarbonyl)phenyl)pent-4-yne-1-sulfonic acid (1.9 g, 6.39 mmol) in dry DCM (30 mL) and dry DMF (2 drops) was treated with oxalyl dichloride (7.0 ml, 83.18 mmol) in 3 portions. The reaction was stirred at rt until judged complete by LCMS analysis. The volatiles were removed in vacuo and the residue was placed under nitrogen and redissolved in dry THF (10 mL) then treated with dimethylamine (2.0 M in THF) (32.0 ml, 63.9 mmol). The reaction mixture was stirred at it until complete by LCMS then partitioned between EtOAc (100 mL) and 10% aq. citric acid (100 mL). The aqueous layer was extracted with EtOAc (2100 mL) before the combined organic extracts were washed sequentially with 10% aq. citric acid ((50 mL), sat. aq. sodium bicarbonate (50 mL), water (50 mL) and brine (50 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was achieved by chromatography (10-50% EtOAc in iso-hexanes) to give methyl 3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoate (1.6 g, 76% yield) as a pale yellow free-flowing oil.

(293) (.sup.1H) DMSO-d.sub.6: 1.90-1.95 (2H, m), 2.61 (2H, t, J=7.1 Hz), 2.79 (6H, s), 3.14-3.22 (2H, m), 3.86 (3H, s), 7.52 (1H, dt, J=0.9, 7.6 Hz), 7.65-7.73 (1H, m), 7.89-7.95 (2H, m) ppm.

(294) MS(ES+) m/z 310 (M+H).

3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoic acid

(295) ##STR00079##

(296) To a solution of methyl 3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoate (1.60 g, 4.86 mmol) in THF (10 mL) was added a solution of lithium hydroxide (0.23 g, 9.72 mmol) in water (6 mL). The reaction mixture was stirred at it until judged complete by LC-MS then the volatiles were removed in vacuo. The residue as diluted with water (10 mL) and acidified to pH 1 with 1 N HCl (aq.). The resulting white precipitate was collected by filtration and washed with water (210 mL), dried by suction for 15 min then in a vacuum oven (40 C.) for 18 h to give 3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoic acid (1.27 g, 87% yield) as a white solid.

(297) (.sup.1H) DMSO-d.sub.6: 1.87-2.02 (2H, m), 2.61 (2H, t, J=7.0 Hz), 2.79 (6H, s), 3.13-3.25 (2H, m), 7.49 (1H, dt, J=0.8, 7.6 Hz), 7.65 (1H, td, J=1.5, 7.7 Hz), 7.86-7.94 (2H, m), 13.16 (1H, s) ppm.

(298) MS(ES+) m/z 296 (M+H).

(R)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzamido)propanoate VSN 81

(299) ##STR00080##

(300) Using the general procedure described for amide coupling, the reaction of 3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzoic acid (0.81 g, 2.74 mmol), (R)-methyl 2-aminopropanoate.HCl (0.421 g, 3.02 mmol), DIPEA (1.20 ml, 6.86 mmol) and HATU (1.20 g, 3.15 mmol) in dry DCM (20 mL) after purification by chromatography (10-100% EtOAc in iso-hexanes) gave the title compound (R)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzamido)propanoate (1.0 g, 93% yield) as a yellow oil

(301) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.96 (2H, dd, J=6.2, 13.9 Hz), 2.62 (2H, t, J=7.1 Hz), 2.79 (6H, s), 3.12-3.24 (2H, m), 3.64 (3H, s), 4.47 (1H, qn, J=7.2 Hz), 7.47 (1H, t, J=7.7 Hz), 7.58 (1H, d, J=7.7 Hz), 7.83 (1H, d, J=7.9 Hz), 7.93 (1H, s), 8.87 (1H, d, J=7.0 Hz) ppm.

(302) (.sup.13C) DMSO-d.sub.6: 16.66, 17.53, 22.20, 37.07, 45.52, 48.28, 51.89, 80.70, 89.86, 122.92, 127.25, 128.76, 130.11, 133.94, 134.07, 165.40, 173.04 ppm.

(303) MS(ES+) m/z 381 (M+H).

(R,Z)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-en-1-yl)benzamido)propanoate VSN 85

(304) ##STR00081##

(305) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-yn-1-yl)benzamido)propanoate (0.50 g, 1.31 mmol) after separation by column chromatography (0-2% MeOH in DCM) then repurification by column chromatography (10% EtOAc in DCM) and prep. HPLC (30% MeCN in water, acidic) gave the title compound (R,Z)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-en-1-yl)benzamido)propanoate (0.13 g, 24% yield @ 95% purity) as a colourless oil.

(306) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.77-1.87 (2H, m), 2.43 (2H, dq, J=1.9, 7.4 Hz), 2.74 (6H, s), 3.03-3.09 (2H, m), 3.65 (3H, s), 4.48 (1H, qn, J=7.3 Hz), 5.75 (1H, td, J=7.2, 11.7 Hz), 6.52 (1H, td, J=1.9, 11.7 Hz), 7.44-7.49 (2H, m), 7.75-7.78 (2H, m), 8.80 (1H, d, J=6.9 Hz) ppm.

(307) (.sup.13C) DMSO-d.sub.6: 16.73, 22.86, 26.78, 37.09, 45.79, 48.29, 51.91, 125.82, 127.68, 128.35, 128.96, 131.35, 132.13, 133.80, 136.94, 166.17, 173.18 ppm.

(308) MS(ES+) m/z 383 (M+H).

(R,Z)-2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-en-1-yl)benzamido)propanoic acid VSN 86

(309) ##STR00082##

(310) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-en-1-yl)benzamido)propanoate (0.075 g, 0.20 mmol) with lithium hydroxide (9.4 mg, 0.39 mmol) gave (R,Z)-2-(3-(5-(N,N-dimethylsulfamoyl)pent-1-en-1-yl)benzamido)propanoic acid (68 mg, 89% yield) as a white solid.

(311) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.4 Hz), 1.75-1.85 (2H, m), 2.39-2.48 (2H, m), 2.75 (6H, s), 3.02-3.11 (2H, m), 4.36-4.47 (1H, m), 5.76 (1H, td, J=7.2, 11.7 Hz), 6.53 (1H, br d, J=11.7 Hz), 7.44-7.51 (2H, m), 7.76-7.79 (2H, m), 8.70 (1H, d, J=7.2 Hz), 12.57 (1H, br s) ppm.

(312) (.sup.13C) DMSO-d.sub.6: 16.89, 22.87, 26.81, 37.11, 45.79, 48.18, 125.81, 127.69, 128.32, 129.02, 131.24, 132.09, 134.06, 136.91, 166.06, 174.25 ppm.

(313) MS(ES+) m/z 369 (M+H).

(314) Synthesis of VSN 82, 87 and 88

methyl 3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoate

(315) ##STR00083##

(316) A suspension of methyl 3-(5-((methylsulfonyl)oxy)pent-1-yn-1-yl)benzoate (1.15 g, 3.88 mmol), pyridin-2(1H)-one (0.41 g, 4.27 mmol) and potassium carbonate (1.07 g, 7.76 mmol) in dry MeCN (12 mL) under nitrogen was heated to 60 C. for 18 h. Then the mixture was allowed to cool to rt before it was partitioned between EtOAc (50 mL) and water (50 mL). The aqueous layer was extracted with EtOAc (230 mL) and the combined organic extracts were washed with water (30 mL) and brine (30 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was achieved by chromatography (0-100% EtOAc in iso-hexanes) to give the title product methyl 3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoate (0.63 g, 53.9% yield) as a clear colourless oil.

(317) (.sup.1H) DMSO-d.sub.6: 1.93 (2H, qn, J=7.1 Hz), 2.47 (2H, t, J=7.1 Hz), 3.86 (3H, s), 3.97-4.04 (2H, m), 6.21 (1H, dt, J=1.4, 6.7 Hz), 6.35-6.40 (1H, m), 7.39 (1H, ddd, J=2.1, 6.6, 8.9 Hz), 7.47-7.54 (1H, m), 7.66 (1H, td, J=1.5, 7.8 Hz), 7.69 (1H, ddd, J=0.7, 2.1, 6.8 Hz), 7.88-7.91 (2H, m) ppm.

(318) (.sup.13C) DMSO-d.sub.6: 16.08, 27.39, 30.65, 47.98, 52.30, 79.85, 90.70, 105.18, 119.62, 123.62, 128.50, 129.15, 129.97, 131.74, 135.77, 139.18, 139.84, 161.45, 165.54 ppm.

(319) MS(ES+) m/z 296 (M+H).

3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoic acid

(320) ##STR00084##

(321) To a solution of methyl 3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoate (0.63 g, 2.13 mmol) in THF (10 mL) was added a solution of lithium hydroxide (0.10 g, 4.27 mmol) in water (3.0 mL). The resulting mixture was stirred at rt for 16 h before the volatiles were removed in vacuo. The residue was then partitioned between 1 N HCl (30 mL) and EtOAc (50 mL) and the aqueous layer was extracted with EtOAc (240 mL). The combined organic extracts were washed with water (30 mL) and brine (30 mL) then dried (MgSO.sub.4), filtered and concentrated in vauco to give the title compound 3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoic acid (0.6 g, 98% yield) as an off-white solid.

(322) (.sup.1H) DMSO-d.sub.6: 1.93 (2H, qn, J=7.0 Hz), 2.43-2.49 (2H, m), 4.01 (2H, t, J=7.1 Hz), 6.21 (1H, dt, J=1.3, 6.7 Hz), 6.34-6.42 (1H, m), 7.39 (1H, ddd, J=2.1, 6.6, 8.9 Hz), 7.45-7.52 (1H, m), 7.63 (1H, td, J=1.4, 7.7 Hz), 7.69 (1H, dd, J=1.6, 6.7 Hz), 7.85-7.93 (2H, m), 13.15 (1H, s) ppm.

(323) MS(ES+) m/z 282 (M+H).

(R)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzamido)propanoate VSN 82

(324) ##STR00085##

(325) Following the general procedure described for amide coupling, the reaction of 3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzoic acid (0.60 g, 2.13 mmol), (R)-methyl 2-aminopropanoate.HCl (0.30 g, 2.13 mmol), DIPEA (0.37 ml, 2.13 mmol) and HATU (0.81 g, 2.13 mmol) in dry DCM (10 mL) after purification by chromatography (1-5% MeOH in DCM) gave (R)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzamido)propanoate (0.43 g, 53.4% yield) as a viscous colourless oil.

(326) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.93 (2H, qn, J=7.1 Hz), 2.44-2.50 (2H, m), 3.64 (3H, s), 3.97-4.05 (2H, m), 4.47 (1H, qn, J=7.2 Hz), 6.22 (1H, dt, J=1.4, 6.7 Hz), 6.36-6.40 (1H, m), 7.40 (1H, ddd, J=2.1, 6.6, 8.9 Hz), 7.46 (1H, t, J=7.8 Hz), 7.57 (1H, td, J=1.3, 7.7 Hz), 7.66-7.73 (1H, m), 7.82 (1H, td, J=1.5, 7.8 Hz), 7.92 (1H, t, J=1.5 Hz), 8.87 (1H, d, J=6.9 Hz) ppm.

(327) (.sup.13C) DMSO-d.sub.6: 16.05, 16.66, 27.51, 47.99, 48.29, 51.89, 80.35, 90.21, 105.23, 119.63, 123.10, 127.13, 128.68, 130.11, 133.90, 134.09, 139.14, 139.88, 161.44, 165.43, 173.05 ppm.

(328) MS(ES+) m/z 367 (M+H).

(R,Z)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-en-1-yl)benzamido)propanoate VSN 87

(329) ##STR00086##

(330) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-yn-1-yl)benzamido)propanoate (0.30 g, 0.82 mmol) gave the named product along with the fully saturated product (30%) (determined by .sup.1H NMR). Separation by column chromatography (EtOAc) and further purification by prep. HPLC (25% MeCN/water, acidic) gave the title compound (R,Z)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-en-1-yl)benzamido)propanoate (87 mg, 28% yield) as a colourless gum. The other component was not isolated.

(331) (.sup.1H) DMSO-d.sub.6: 1.41 (3H, d, J=7.3 Hz), 1.79 (2H, qn, J=7.5 Hz), 2.23-2.38 (2H, m), 3.65 (3H, s), 3.83-3.92 (2H, m), 4.49 (1H, qn, J=7.3 Hz), 5.77 (1H, td, J=7.3, 11.7 Hz), 6.14 (1H, dt, J=1.4, 6.7 Hz), 6.33-6.38 (1H, m), 6.51 (1H, br d, J=11.7 Hz), 7.38 (1H, ddd, J=2.1, 6.6, 8.9 Hz), 7.41-7.50 (2H, m), 7.62 (1H, dd, J=1.6, 6.8 Hz), 7.76 (2H, dd, J=1.9, 3.8 Hz), 8.84 (1H, d, J=6.9 Hz) ppm.

(332) (.sup.13C) DMSO-d.sub.6: 16.76, 25.23, 28.71, 48.16, 48.31, 51.94, 105.14, 119.57, 125.83, 127.67, 128.32, 128.69, 131.30, 132.46, 133.82, 136.97, 139.11, 139.84, 161.38, 166.20, 173.22 ppm.

(333) MS(ES+) m/z 369 (M+H).

R,Z)-2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-en-1-yl)benzamido)propanoic acid VSN 88

(334) ##STR00087##

(335) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-en-1-yl)benzamido)propanoate (40 mg, 0.11 mmol) with lithium hydroxide (5.2 mg, 0.22 mmol) gave (R,Z)-2-(3-(5-(2-oxopyridin-1(2H)-yl)pent-1-en-1-yl)benzamido)propanoic acid (30 mg, 76% yield) as a white solid.

(336) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.4 Hz), 1.78 (2H, qn, J=7.5 Hz), 2.25-2.36 (2H, m), 3.83-3.92 (2H, m), 4.42 (1H, qn, J=7.5 Hz), 5.76 (1H, td, J=7.3, 11.7 Hz), 6.13 (1H, dt, J=1.4, 6.7 Hz), 6.32-6.37 (1H, m), 6.50 (1H, br d, J=11.6 Hz), 7.37 (1H, ddd, J=2.1, 6.6, 9.0 Hz), 7.40-7.48 (2H, m), 7.60 (1H, dd, J=1.5, 6.8 Hz), 7.74-7.76 (2H, m), 8.67 (1H, d, J=7.3 Hz), 12.47 (1H, s) ppm.

(337) (.sup.13C) DMSO-d.sub.6: 16.89, 25.19, 28.66, 48.12, 48.17, 105.08, 119.53, 125.74, 127.60, 128.21, 128.69, 131.11, 132.35, 134.09, 136.89, 139.04, 139.76, 161.34, 166.03, 174.21 ppm.

(338) MS(ES+) m/z 355 (M+H).

(339) Synthesis of VSN 83, 89 and 90

methyl 3-(5-(methylamino)pent-1-yn-1-yl)benzoate

(340) ##STR00088##

(341) To a solution of methyl 3-(5-((methylsulfonyl)oxy)pent-1-yn-1-yl)benzoate (2.40 g, 8.10 mmol) in dry THF (25.0 mL) was added methanamine (40 wt % in water) (7.01 ml, 81 mmol). The reaction was then stirred at 45 C. until judged complete by LCMS analysis. The reaction mixture was then partitioned between EtOAc (150 mL) and sat. aq. sodium bicarbonate (100 mL) and the aqueous layer was extracted with EtOAc (2100 mL). The combined organic extracts were washed with water (3100 mL) and brine (50 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo to give the title product methyl 3-(5-(methylamino)pent-1-yn-1-yl)benzoate (2.0 g, 70.5% yield, 66% purity) as a 2:1 mixture with N-methyl-3-(5-(methylamino)pent-1-yn-1-yl)benzamide. No further purification attempted.

(342) MS(ES+) m/z 232 (M+H).

methyl 3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoate

(343) ##STR00089##

(344) A solution of methyl 3-(5-(methylamino)pent-1-yn-1-yl)benzoate (1.05 g, 4.54 mmol) (66% purity) and DIPEA (1.59 ml, 9.08 mmol) in dry DCM (10 mL) was treated with acetyl chloride (0.48 ml, 6.81 mmol). The reaction mixture was stirred at rt until judged complete by LCMS analysis. The volatiles were then removed in vacuo and the residue was partitioned between EtOAc (50 mL) and 1 N HCl (25 mL) and the aqueous layer was extracted with EtOAc (230 mL). The combined organic layers were washed with sat. aq. NaHCO.sub.3 (25 mL), water (25 mL) and brine (25 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo to give methyl 3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoate (1.20 g, 63.8% yield) as a 2:1 mixture with N-methyl-3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzamide.

(345) MS(ES+) m/z 274 (M+H).

3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoic acid

(346) ##STR00090##

(347) To a solution of methyl 3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoate (1.20 g, 4.39 mmol) in THF (15 mL) was added a solution of lithium hydroxide (0.210 g, 8.78 mmol) in water (3.0 mL). The reaction mixture was stirred at rt until judged complete by LCMS. The volatiles were removed in vacuo and the residue was partitioned between EtOAc (20 mL) and water (20 mL). The aqueous layer was extracted with with EtOAc (320 mL) then acidified to pH 1 with 1 N HCl and extracted with EtOAc (320 mL). The combined organic extracts were washed with water (20 mL) and brine (20 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo to give 3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoic acid (0.68 g, 56.7% yield) as a viscous brown oil.

(348) (.sup.1H) DMSO-d.sub.6: 1.67-1.87 (2H, m), 1.91 (1.5H, s), 1.98 (1.5H, s), 2.41 (1H, t, J=7.1 Hz), 2.45-2.49 (1H, m), 2.80 (1.5H, s), 2.97 (1.5H, s), 3.34-3.46 (2H, m), 7.44-7.53 (1H, m), 7.59-7.66 (1H, m), 7.85-7.94 (2H, m), 12.79 (1H, s) ppm (compound rotomeric hence some resonances are split).

(349) MS(ES+) m/z 260 (M+H).

(R)-methyl 2-(3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzamido)propanoate VSN 83

(350) ##STR00091##

(351) Following the general procedure described for amide coupling, the reaction of 3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzoic acid (0.68 g, 2.49 mmol), (R)-methyl 2-aminopropanoate.HCl (0.35 g, 2.49 mmol), HATU (1.14 g, 2.99 mmol) and DIPEA (1.33 ml, 7.47 mmol) in dry DCM (10 mL) after purification by chromatography (1-3% MeOH in DCM) gave (R)-methyl 2-(3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzamido)propanoate (0.40 g, 45.7% yield) as a pale yellow oil.

(352) (.sup.1H) DMSO-d.sub.6 (@ 100 C.): 1.43 (3H, d, J=7.3 Hz), 1.77-1.87 (2H, m), 2.01 (3H, s), 2.43-2.50 (2H, m), 2.94 (3H, s), 3.39-3.48 (2H, m), 3.68 (3H, s), 4.54 (1H, qn, J=7.2 Hz), 7.43 (1H, t, J=7.7 Hz), 7.53 (1H, td, J=1.4, 7.7 Hz), 7.81 (1H, td, J=1.5, 7.8 Hz), 7.89 (1H, t, J=1.5 Hz), 8.50 (1H, br s) ppm.

(353) (.sup.13C) DMSO-d.sub.6: 15.90, 16.30, 16.66, 21.06, 21.72, 26.07, 26.74, 32.47, 35.81, 45.97, 48.28, 48.82, 51.89, 79.95, 80.31, 90.57, 90.98, 123.07, 123.23, 127.07, 127.13, 128.73, 128.76, 130.01, 130.06, 133.93, 133.95, 134.00, 165.42, 169.31, 169.66, 173.05 ppm. (Note: some peaks are doubled as compound is rotomeric).

(354) MS(ES+) m/z 345 (M+H).

(R,Z)-methyl 2-(3-(5-(N-methylacetamido)pent-1-en-1-yl)benzamido)propanoate VSN 89

(355) ##STR00092##

(356) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(5-(N-methylacetamido)pent-1-yn-1-yl)benzamido)propanoate (0.32 g, 0.93 mmol) after separation by column chromatography (0-2% MeOH in DCM) and preparative HPLC (20-30% MeCN, acidic) gave the title compound (R,Z)-methyl 2-(3-(5-(N-methylacetamido)pent-1-en-1-yl)benzamido)propanoate (0.16 g, 48% yield) as a colourless oil.

(357) (.sup.1H) DMSO-d.sub.6 (@ 100 C.): 1.44 (3H, d, J=7.3 Hz), 1.63-1.75 (2H, m), 1.95 (3H, s), 2.24-2.35 (2H, m), 2.95 (3H, s), 3.25-3.33 (2H, m), 3.68 (3H, s), 4.55 (1H, qn, J=7.2 Hz), 5.77 (1H, td, J=7.3, 11.7 Hz), 6.49 (1H, d, J=11.7 Hz), 7.41-7.49 (2H, m), 7.70-7.80 (2H, m), 8.42 (1H, d, J=5.9 Hz) ppm.

(358) (.sup.13C) DMSO-d.sub.6: 16.73, 21.04, 21.70, 25.34, 25.54, 26.97, 27.85, 32.53, 35.62, 46.17, 48.30, 49.40, 51.92, 125.73, 125.79, 127.58, 127.61, 128.31, 128.34, 128.60, 131.36, 131.42, 132.77, 133.04, 133.77, 133.79, 137.02, 137.09, 166.18, 166.22, 169.23, 169.50, 173.20 ppm (Note: some peaks are doubled as compound is rotomeric).

(359) MS(ES+) m/z 347 (M+H).

(R,Z)-2-(3-(5-(N-methylacetamido)pent-1-en-1-yl)benzamido)propanoic acid VSN 90

(360) ##STR00093##

(361) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(5-(N-methylacetamido)pent-1-en-1-yl)benzamido)propanoate (0.10 g, 0.289 mmol) with lithium hydroxide (14 mg, 0.58 mmol) gave (R,Z)-2-(3-(5-(N-methylacetamido)pent-1-en-1-yl)benzamido)propanoic acid (85 mg, 86% yield) as a colourless gum.

(362) (.sup.1H) DMSO-d.sub.6 (@100 C.): 1.43 (3H, d, J=7.3 Hz), 1.67 (2H, m), 1.95 (3H, s), 2.24-2.35 (2H, m), 2.86 (3H, br s), 3.26-3.34 (2H, m), 4.49 (1H, qn, J=7.3 Hz), 5.77 (1H, td, J=7.3, 11.7 Hz), 6.49 (1H, br d, J=11.6 Hz), 7.40-7.48 (2H, m), 7.71-7.79 (2H, m), 8.26 (1H, m) ppm. Note: No OH observed.

(363) (.sup.13C) DMSO-d.sub.6: 16.88, 21.05, 21.71, 25.35, 25.54, 26.98, 27.85, 32.55, 35.64, 46.19, 48.17, 49.41, 125.71, 125.77, 127.57, 127.60, 128.26, 128.29, 128.34, 128.64, 131.23, 131.30, 132.71, 132.99, 134.04, 134.05, 136.97, 137.05, 166.07, 166.11, 169.23, 169.50, 174.24 ppm (Note: some peaks are doubled as compound is rotomeric).

(364) MS(ES+) m/z 333 (M+H).

(365) Synthesis of VSN 84, 91 and 92

methyl 3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoate

(366) ##STR00094##

(367) A solution of methyl 3-(5-(methylamino)pent-1-yn-1-yl)benzoate (1.0 g, 4.32 mmol) (66% purity), DIPEA (1.13 ml, 6.49 mmol) in dry DCM (10 mL) was treated with methanesulfonyl chloride (0.42 ml, 5.40 mmol). The reaction mixture was stirred at rt until judged complete by LCMS analysis. The reaction mixture was partitioned between DCM (50 mL) and 1 N HCl (25 mL). The aq. layer was extracted with DCM (230 mL) before the combined organic extracts were washed with water (25 mL) and brine (25 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo to give methyl 3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoate (1.20 g, 59.2% yield) as a 2:1 mixture with N-methyl-3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzamide. Carried forward as a mixture.

(368) MS(ES+) m/z 310 (M+H).

3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoic acid

(369) ##STR00095##

(370) To a solution of methyl 3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoate (1.20 g, 3.88 mmol) in THF (15 mL) was added a solution of lithium hydroxide (0.19 g, 7.76 mmol) in water (3.0 mL) The reaction mixture was stirred at rt until judged complete by LCMS. The volatiles were removed in vacuo and the residue was partitioned between EtOAc (20 mL) and water (20 mL). The aq. layer was acidified to pH 1 with 1 N HCl and extracted with EtOAc (330 mL). The combined organic extracts were washed with water (30 mL) and brine (30 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo to give 3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoic acid (0.50 g, 42.8% yield) as a pale brown solid.

(371) (.sup.1H) DMSO-d.sub.6: 1.81 (2H, qn, J=7.1 Hz), 2.44-2.49 (2H, m), 2.77 (3H, s), 2.88 (3H, s), 3.14-3.22 (2H, m), 7.45-7.52 (1H, m), 7.63 (1H, td, J=1.4, 7.7 Hz), 7.85-7.93 (2H, m), 13.13 (1H, s) ppm

(372) MS(ES+) m/z 296 (M+H).

(R)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzamido)propanoate VSN 84

(373) ##STR00096##

(374) Following the general procedure described for amide coupling, the reaction of 3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzoic acid (0.50 g, 1.69 mmol), (R)-methyl 2-aminopropanoate.HCl (0.26 g, 1.86 mmol), HATU (0.74 g, 1.95 mmol) and DIPEA (0.739 ml, 4.23 mmol) in dry DCM (10 mL) after purification by chromatography (30-70% EtOAc in iso-hexanes) gave (R)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzamido)propanoate (0.39 g, 57.5% yield) as a pale yellow oil.

(375) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.82 (2H, qn, J=7.1 Hz), 2.46-2.49 (2H, m), 2.77 (3H, s), 2.88 (3H, s), 3.18 (2H, t, J=7.0 Hz), 3.64 (3H, s), 4.47 (1H, qn, J=7.1 Hz), 7.46 (1H, t, J=7.8 Hz), 7.56 (1H, td, J=1.3, 7.7 Hz), 7.82 (1H, td, J=1.4, 7.8 Hz), 7.91 (1H, t, J=1.5 Hz), 8.86 (1H, d, J=6.9 Hz) ppm.

(376) (.sup.13C) DMSO-d.sub.6: 15.92, 16.66, 26.59, 34.54, 34.57, 48.28, 48.60, 51.90, 80.18, 90.62, 123.17, 127.09, 128.73, 130.04, 133.93, 134.04, 165.43, 173.05 ppm.

(377) MS(ES+) m/z 381 (M+H).

(R,Z)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-en-1-yl)benzamido)propanoate VSN 91

(378) ##STR00097##

(379) Following the general procedure for the Lindlar reduction, the hydrogenation of (R)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-yn-1-yl)benzamido)propanoate (0.32 g, 0.84 mmol) gave the named product along with the trans double bond isomer (<5%) and fully saturated product (10%) (determined by .sup.1H NMR). Separation by column chromatography (10% EtOAc in DCM) gave the title compound (R,Z)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-en-1-yl)benzamido)propanoate (0.19 g, 59% yield) as a colourless oil. The other 2 components were not isolated.

(380) (.sup.1H) DMSO-d.sub.6: 1.40 (3H, d, J=7.3 Hz), 1.68 (2H, qn, J=7.1 Hz), 2.26-2.35 (2H, m), 2.72 (3H, s), 2.83 (3H, s), 3.03-3.07 (2H, m), 3.64 (3H, s), 4.48 (1H, qn, J=7.2 Hz), 5.77 (1H, td, J=7.3, 11.8 Hz), 6.50 (1H, td, J=1.8, 11.8 Hz), 7.43-7.51 (2H, m), 7.73-7.80 (2H, m), 8.80 (1H, d, J=6.9 Hz) ppm. Note: contaminated with 5-10% trans isomer.

(381) (.sup.13C) DMSO-d.sub.6: 16.71, 25.23, 27.48, 34.41, 34.48, 48.26, 49.04, 51.87, 125.72, 127.60, 128.27, 128.52, 131.29, 132.66, 133.79, 137.02, 166.16, 173.14 ppm.

(382) MS(ES+) m/z 383 (M+H).

(R,Z)-2-(3-(5-(N-methylmethylsulfonamido)pent-1-en-1-yl)benzamido)propanoic acid VSN 92

(383) ##STR00098##

(384) Following the general procedure for saponification, the reaction of (R,Z)-methyl 2-(3-(5-(N-methylmethylsulfonamido)pent-1-en-1-yl)benzamido)propanoate (0.11 g, 0.288 mmol) with lithium hydroxide (14 mg, 0.58 mmol) gave (R,Z)-2-(3-(5-(N-methylmethylsulfonamido)pent-1-en-1-yl)benzamido)propanoic acid (0.1 g, 90% yield) as a white solid. Note: 5% isomerisation to trans double bond under reaction conditions observed.

(385) (.sup.1H) DMSO-d.sub.6: 1.43 (3H, d, J=7.3 Hz), 1.72 (2H, qn, J=7.4 Hz), 2.33 (2H, dq, J=1.8, 7.5 Hz), 2.75 (3H, s), 2.81 (3H, s), 3.08-3.14 (2H, m), 4.49 (1H, qn, J=7.3 Hz), 5.78 (1H, td, J=7.3, 11.7 Hz), 6.50 (1H, dd, J=11.7, 1.6 Hz), 7.42-7.47 (2H, m), 7.72-7.78 (2H, m), 8.26 (1H, d, J=6.9 Hz), 12.04 (1H, br s) ppm.

(386) (.sup.13C) DMSO-d.sub.6: 16.88, 25.27, 27.52, 34.46, 34.48, 48.17, 49.07, 125.74, 127.63, 128.27, 128.59, 131.21, 132.65, 134.05, 137.00, 166.08, 174.24 ppm.

(387) MS(ES+) m/z 369 (M+H).

(388) In Vitro Activity

(389) Compounds were evaluated using a mouse vas deferens preparation [Ward S, Mastriani D, Casiano F and Arnold R (1990) J Pharmacol Exp Ther 255:1230-1239]. The mouse vas deferens isolated tissue preparation is used as an in vitro bioassay to assess the actions of compounds on neurotransmission. Electrically-evoked contractions are induced in the preparation and compounds which affect neurotransmission will inhibit these contractions in a concentration related manner. The vas deferens has been widely used to assess the effects of compounds at the cannabinoid CB1 receptor, which is located pre-synaptically. CB1 agonists inhibit electrically-evoked contractions.

(390) Methods

(391) Vasa deferentia were obtained from albino MF1 mice weighing 31-59 g. The tissues were mounted vertically in 4 ml organ baths. They were then subjected to electrical stimulation of progressively greater intensity, followed by an equilibration procedure in which they were exposed to alternate periods of stimulation (2 min) and rest (10 min) until contractions with consistent amplitudes were obtained. These contractions were monophasic and isometric, and were evoked by 0.5 s trains of pulses of 110% maximal voltage (train frequency 0.1 Hz; pulse frequency 5 Hz; pulse duration 0.5 ms).

(392) All drug additions were made to the organ baths after the equilibration period and there was no washout between these additions. Potential antagonist or its vehicle were added to the preparation. This was followed 28 min later by a 2-min period of electrical stimulation, at the end of which the lowest of a series of concentrations of the twitch inhibitors was applied. After a period of rest, the tissues were electrically stimulated for 2 min and then subjected to a further addition of twitch inhibitor. This cycle of drug addition, rest and 2 min stimulation was repeated so as to construct cumulative concentrationresponse curves. Only one concentrationresponse curve was constructed per tissue.

(393) Analysis of Data

(394) Inhibition of the electrically-evoked twitch response of the vas deferens has been expressed in percentage terms, and this has been calculated by comparing the amplitude of the twitch response after each addition of a twitch inhibitor with its amplitude immediately before the first addition of the inhibitor. Values for EC.sub.50, maximal effect (E.sub.max) and the s.e.m. or 95% confidence limits of these values have been calculated by nonlinear regression analysis using the equation for a sigmoid concentrationresponse curve (GraphPad Prism).

(395) In Vivo Peripheral CB.sub.1 Receptor Activation

(396) Assessment of Spasticity

(397) Further studies were undertaken using cannabinoid knockout mice, including CB.sub.1, CB.sub.2, VR-1, FAAH and conditional CB.sub.1 knockout mice. Spasticity may be induced in ABH (significant spasticity occurs in 50-60% of animals in 80 days after 3-4 disease episodes.sup.1) or ABH.CB.sub.1/ (significant spasticity occurs in 80-100% of animals in 30-40 days after 1-2 disease episodes). Compounds are injected initially intravenously (to limit first pass effects), i.p. or orally. Spasticity is assessed (n=6-7/group) by resistance to hindlimb flexion using a strain gauge [Baker, D. et al, Nature 2000, 404, 84-87]. Animals serve as their own controls and will be analysed in a pairwise fashion. To reduce the number of animals, effort and expense, following a drug-free period (spasticity returns within 24 h) these animals receive different doses and or vehicle. Low doses of CB.sub.1 agonists and CNS active CP55,940, as control, are locally (subcutaneous, intra-muscularly) administered into spastic ABH mice and the lack of activity in a contralateral limb analysed [Fox, A. et al, Pain 2001, 92, 91-100]. Expression of CB.sub.1 in the peripheral nervous system, including dorsal root ganglia, a non-CNS site for CB-mediated nociception can be removed using peripherin-Cre transgenic mouse [Zhou, L. et al, FEBS Lett. 2002, 523, 68-72]. These conditional KO mice are maintained on the C57BL/6 background. These mice develop EAE following induction with myelin oligodendrocyte glycoprotein residues 35-55 peptide [Amor, S. et al, J. Immunol. 1994, 153, 4349-4356].

(398) Results

(399) The compounds of the invention inhibited electrically-evoked contractions of the mouse isolated vas deferens in a similar manner to the standard CB1 receptor agonist, WIN55212-2 (Table 1, FIG. 1). Table 1 shows that the potency (EC.sub.50) of VSN44 is unexpectedly ten-fold greater than for VSN16R (1.42 nM for VSN-44 cf 10.44 nM for VSN-16R). Further results for the vas deferens assay are shown in Table 2 for selected compounds according to the invention.

(400) A VSN-16R dose response curve in mouse vas deferens shows that the EC.sub.50 stays relatively constant across wild type (10.44 nM), CB.sub.1 knock-out (12.66 nM) and CB.sub.2 knock-out (17.61 nM); see FIG. 3.

(401) FIG. 2 shows the effect of VNS-44 on spasticity (percentage spasticity vs baseline) against time after administration.

CONCLUSIONS

(402) The compounds tested are potent inhibitors of electrically-evoked contractions of the mouse isolated vas deferens. Advantageously, VSN-44 shows at least a 10-fold increase in potency over VSN-16R in the mouse vas deferens study.

(403) Various modifications and variations of the described methods of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, various modifications of the described modes for carrying out the invention which are obvious to those skilled in chemistry or related fields are intended to be within the scope of the following claims.

(404) TABLE-US-00001 TABLE 1 Emax, pEC.sub.50 SEM and EC.sub.50 values for compound [1] (VSN-44) Emax (%) 95% confidence Compound limits pEC.sub.50 SEM EC.sub.50 (nM) WIN55212-2 62 (52-72) 8.41 0.237 3.86 VSN-22R 43 (28-56) 8.03 0.367 9.42 VSN-44 46 (34-58) 8.85 0.561 1.42 VSN16R 56 (48-64) 7.98 0.367 10.44

(405) TABLE-US-00002 TABLE 2 Emax, and EC.sub.50 values for compound selected compounds according to the invention Emax EC50 EC50 Emax 95% Cpd Structure Mol Wt (nM) 95% Cls (%) Cls VSN 16R 318.41952 0.7216 0.1055-4.94 40.71 30.1-51.33 [1] VSN 44R 00 332.40298 0.006546 0.002807-0.01527 64.02 56.83-71.21 [20] 01 370.45237 0.4662 0.0676-3.2 44.73 33.83-55.63 [21] 02 372.46831 0.1617 0.04544-0.5754 58.31 52.45-64.18 [22] 03 330.38704 0.1116 0.0333-0.3744 52.9 45.06-60.73 [23] 04 332.40298 0.02247 0.007191-0.0702 59.25 52.43-66.07 [15] 05 318.37589 0.1024 0.04556-0.23 53.71 48.18-59.23 [24] 06 360.41353 0.4122 0.1221-1.39 51.53 43.42-59.64 [25] 07 362.42947 0.3591 0.07067-1.83 52.86 41.81-63.9 [18] 08 348.40238 0.3822 0.0931-1.57 62.68 51.72-73.64 [27] 09 422.52885 0.09083 0.02804-0.2942 55.3 47.37-63.24 [17] 0 408.50176 0.1397 0.03319 56.28 46.09-66.46 [29] 362.42947 0.04555 0.01725-0.1203 62 55.16-68.84 [14] 348.40238 0.1244 0.04403-0.3513 58.47 50.49-66.44 [30] 420.51291 0.06135 0.02416-0.1558 46.32 40.88-51.76 [31] 422.52885 0.01224 0.002949-0.05084 46.99 38.74-55.24 [16] 408.50176 0.00604 0.003016-0.0121 73.88 68.35-79.4 [32] 344.41413 0.01312 0.002865-0.06007 54.57 45.07-64.07 [33] 348.446 [4] 334.4189 [34] 330.387 [35] 0 334.4189 [36] 320.3918 [37] 344.4141 [38] 348.446 [11] 334.4189 [39] 372.4683 [40] 376.5002 [13] 362.4731 [41] 344.4141 [42] 346.4301 [5] 0 332.403 [43] 298.3446 [44] 300.3606 [6] 286.3335 [45] 380.4664 [46] 366.4205 [47] 344.4141 [48] 380.4664 [49] 382.4823 [3] 368.4552 [50] 0 368.4364 [51] 354.4093 [52] 346.4301 [53] 332.403 [54] 382.4823 [55] 368.4552