ANTIMICROBIAL AGENTS

Abstract

The invention provides novel analogues of enacyloxin Ha and their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs. Such compounds are effective in the treatment of infections caused by Gram-negative bacteria such as Acinetobacter baumannii. Compounds in accordance with the invention include those of formula (A), and their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs: In formula (A): X is 0 or NR.sup.x (where R* is either H or C.sub.1-3 alkyl, e.g. CH.sub.3); R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. a C.sub.1-6 alkyl group); Y is one of the following groups: (wherein each * denotes the point of attachment of the group to the remainder of the molecule; R.sup.9 is H, F, Cl, Br or I; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are =0, preferably R.sup.4 is H and R.sup.5 is OH; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H and R.sup.7 is OH, or R.sup.7 and R.sup.7 together are =0, preferably R7 is H and R7 is OH); and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

Claims

1. A compound of formula (A), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00116## wherein: X is O or NR.sup.x (where R.sup.x is either H or C.sub.1-3 alkyl, e.g. CH.sub.3); R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. a C.sub.1-6 alkyl group); Y is one of the following groups: ##STR00117## (where each * denotes the point of attachment of the group to the remainder of the molecule); R.sup.9 is H, F, Cl, Br or I; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O, preferably R.sup.4 is H and R.sup.5 is OH; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H and R.sup.7 is OH, or R.sup.7 and R.sup.7 together are O, preferably R.sup.7 is H and R.sup.7 is OH; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

2. A compound as claimed in claim 1 of formula (IV), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00118## wherein: X is O or NR.sup.x (where R.sup.x is either H or C.sub.1-3 alkyl, e.g. CH.sub.3); R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O, preferably R.sup.4 is H and R.sup.5 is OH; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H and R.sup.7 is OH, or R.sup.7 and R.sup.7 together are O, preferably R.sup.7 is H and R.sup.7 is OH; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. C.sub.1-6 alkyl group); and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

3. A compound as claimed in claim 2, wherein R.sup.1 is an optionally substituted cyclohexyl or cyclopentyl ring, an optionally substituted cyclohexenyl ring, or an optionally substituted straight-chained C.sub.1-6 alkyl group.

4. A compound as claimed in claim 2 or claim 3, wherein R.sup.1 is substituted by one or more of the following groups: OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), C.sub.1-3 alkyl (e.g. CH.sub.3), CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) and SO.sub.3H.sub.2 (or an ester thereof).

5. A compound as claimed in any one of claims 2 to 4, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

6. A compound as claimed in claim 5, wherein R.sup.8 is ethyl.

7. A compound as claimed in claim 2 of formula (IVb) or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00119## wherein: X is as defined in claim 2; R.sup.d is H, OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), or C.sub.1-3 alkyl (e.g. CH.sub.3), preferably R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3; R.sup.e is H, CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) or SO.sub.3H.sub.2 (or an ester thereof), preferably R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2; R.sup.2, R.sup.3 and R.sup.6 are as defined in claim 2; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

8. A compound as claimed in claim 7, wherein: X is O or NH, preferably O; R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3, preferably OH; R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2, preferably CO.sub.2H; R.sup.2 is H, F, Cl, Br, I or CH.sub.3, preferably H, Cl or Br, e.g. H or Cl; R.sup.3 is H or OH; R.sup.6 is H, Cl or Br, e.g. R.sup.6 is H or Cl; and each of C.sub.2-C.sub.3, C.sub.4-C.sub.0, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds.

9. A compound as claimed in any one of claims 2 to 8, wherein X is O.

10. A compound as claimed in any one of claims 2 to 9, wherein R.sup.2 is Cl or H, preferably Cl, or wherein R.sup.2 is Br.

11. A compound as claimed in any one of claims 2 to 10, wherein R.sup.3 is OH.

12. A compound as claimed in any one of claims 2 to 11, wherein R.sup.4 is H and R.sup.5 is OH.

13. A compound as claimed in any one of claims 2 to 12, wherein R.sup.6 is H or Cl, preferably H.

14. A compound as claimed in any one of claims 2 to 13, wherein R.sup.7 is H and R.sup.7 is OH.

15. A compound as claimed in any one of claims 2 to 14, wherein each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

16. A compound as claimed in claim 2 of formula (IVc), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00120## wherein X, R.sup.d, R.sup.e, R.sup.2, R.sup.3 and R.sup.6 are as defined in any one of claims 2, 7 to 11 and 13.

17. A compound as claimed in claim 2 of formula (IVd), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00121## wherein X, R.sup.d, R.sup.e, R.sup.2, R.sup.3 and R.sup.6 are as defined in any one of claims 2, 7 to 11 and 13.

18. A compound as claimed in claim 2 selected from any of the following compounds, and their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs: ##STR00122##

19. A compound as claimed in claim 2 selected from any of the following compounds, and their pharmaceutically acceptable salts, metabolites, and prodrugs: ##STR00123##

20. A compound as claimed in claim 1 of formula (I), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00124## wherein: X is O or NR.sup.x (where R.sup.x is either H or C.sub.1-3 alkyl, e.g. CH.sub.3); R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.8 is a straight-chained or branched C.sub.18 alkyl group (e.g. a C.sub.1-6 alkyl group); R.sup.9 is H, F, Cl, Br or I, preferably H or Cl; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

21. A compound as claimed in claim 20, wherein R.sup.1 is an optionally substituted cyclohexyl or cyclopentyl ring, an optionally substituted cyclohexenyl ring, or an optionally substituted straight-chained C.sub.1-6 alkyl group.

22. A compound as claimed in claim 20 or claim 21, wherein R.sup.1 is substituted by one or more of the following groups: OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), C.sub.1-3 alkyl (e.g. CH.sub.3), CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) and SO.sub.3H.sub.2 (or an ester thereof).

23. A compound as claimed in any one of claims 20 to 22, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

24. A compound as claimed in claim 23, wherein R.sup.8 is ethyl.

25. A compound as claimed in claim 20 of formula (Ia), or a pharmaceutically acceptable, salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00125## wherein: X is as defined in claim 20; R.sup.d is H, OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), or C.sub.1-3 alkyl (e.g. CH.sub.3), preferably R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3; R.sup.e is H, CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) or SO.sub.3H.sub.2 (or an ester thereof), preferably R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2; R.sup.2, R.sup.3 and R.sup.9 are as defined in claim 20; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

26. A compound as claimed in claim 25, wherein: X=O or NH; R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3, preferably OH; R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2, preferably CO.sub.2H; R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.9 is H, Cl or Br, e.g. R.sup.9 is H or Cl; and each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds.

27. A compound as claimed in any one of claims 20 to 26, wherein X is O.

28. A compound as claimed in any one of claims 20 to 27, wherein R.sup.2 is Cl, or wherein R.sup.2 is Br.

29. A compound as claimed in any one of claims 20 to 28, wherein R.sup.3 is OH.

30. A compound as claimed in any one of claims 20 to 29, wherein each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

31. A compound as claimed in claim 20 of formula (Ib), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00126## wherein X, R.sup.d, R.sup.e, R.sup.3 and R.sup.9 are as defined in any one of claims 20, 25 to 27 and 29.

32. A compound as claimed in claim 20 of formula (Ic), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00127## wherein X, R.sup.d, R.sup.e, R.sup.3 and R.sup.9 are as defined in any one of claims 20, 25 to 27 and 29.

33. A compound as claimed in claim 20 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00128##

34. A compound as claimed in claim 20 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00129##

35. A compound of formula (II), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00130## wherein: R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H, OH, or OC(O)NR.sub.2 (where each R is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), preferably R.sup.7 is H, OH or OC(O)NH.sub.2; R.sup.8 is a straight-chained or branched C.sub.1-3 alkyl group (e.g. C.sub.1-6 alkyl group); R.sup.x is either H or C.sub.1-3 alkyl, e.g. CH.sub.3; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

36. A compound as claimed in claim 35, wherein R.sup.1 is an optionally substituted cyclohexyl or cyclopentyl ring, an optionally substituted cyclohexenyl ring, or an optionally substituted straight-chained C.sub.1-6 alkyl group.

37. A compound as claimed in claim 35 or claim 36, wherein R.sup.1 is substituted by one or more of the following groups: OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), C.sub.1-3 alkyl (e.g. CH.sub.3), CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) and SO.sub.3H.sub.2 (or an ester thereof).

38. A compound as claimed in any one of claims 35 to 37, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

39. A compound as claimed in claim 38, wherein R.sup.8 is ethyl.

40. A compound as claimed in claim 35 of formula (IIa) or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00131## wherein: R.sup.d is H, OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), or C.sub.1-3 alkyl (e.g. CH.sub.3), preferably R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3; R.sup.e is H, CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) or SO.sub.3H.sub.2 (or an ester thereof), preferably R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2; R.sup.2 to R.sup.7, and R.sup.x are as defined in claim 35; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

41. A compound as claimed in claim 40, wherein R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3, preferably H, OH or NH.sub.2; R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2, preferably CO.sub.2H; R.sup.2 is F, Cl, Br or I, preferably Cl; R.sup.3 is OH; R.sup.4 and R.sup.5 together are O; R.sup.6 is F, Cl, Br or I, preferably Cl; R.sup.7 is OC(O)NH.sub.2; R.sup.x is H or CH.sub.3, preferably H; and C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds.

42. A compound as claimed in any one of claims 35 to 41, wherein R.sup.2 is Cl.

43. A compound as claimed in any one of claims 35 to 42, wherein R.sup.3 is OH.

44. A compound as claimed in any one of claims 35 to 43, wherein R.sup.4 and R.sup.5 together are O.

45. A compound as claimed in any one of claims 35 to 44, wherein R.sup.6 is Cl.

46. A compound as claimed in any one of claims 35 to 45, wherein R.sup.7 is OC(O)NH.sub.2.

47. A compound as claimed in any one of claims 35 to 46, wherein each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

48. A compound as claimed in claim 35 of formula (lib), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00132## wherein R.sup.1 to R.sup.7, and R.sup.x are as defined in any one of claims 35 to 37 and 40 to 46.

49. A compound as claimed in claim 25 of formula (IIc), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00133## wherein R.sup.1 to R.sup.7, and R.sup.x are as defined in any one of claims 35 to 37 and 40 to 46.

50. A compound as claimed in claim 35 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00134##

51. A compound as claimed in claim 35 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00135##

52. A compound of formula (IIIa) or (IIIb), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof, or a stereoisomeric form thereof at one or more of positions C6, C11 to C15, and C17 to C19 in the polyketide chain: ##STR00136## wherein: R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H, OH, or OC(O)NR.sub.2 (where each R is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), preferably R.sup.7 is H, OH or OC(O)NH.sub.2; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. C.sub.1-6 alkyl group); and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

53. A compound as claimed in claim 52, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

54. A compound as claimed in claim 53, wherein R.sup.8 is ethyl.

55. A compound as claimed in any one of claims 52 to 54, wherein R.sup.2 is F, Cl, Br or I, preferably Cl.

56. A compound as claimed in any one of claims 52 to 55, wherein R.sup.3 is OH.

57. A compound as claimed in any one of claims 52 to 56, wherein R.sup.4 and R.sup.5 together are O.

58. A compound as claimed in any one of claims 52 to 57, wherein R.sup.6 is Cl.

59. A compound as claimed in any one of claims 52 to 58, wherein R.sup.7 is OC(O)NH.sub.2.

60. A compound as claimed in any one of claims 52 to 59, wherein each of C.sub.2-C.sub.3, C.sub.3-C.sub.4, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

61. A compound as claimed in claim 52 of formula (IIIc) or (IIId), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00137## wherein R.sup.2 to R.sup.7 are as defined in any one of claims 52 and 55 to 59.

62. A compound as claimed in claim 52 of formula (IIIe) or (IIIf), or a pharmaceutically acceptable salt, metabolite or prodrug thereof: ##STR00138## wherein R.sup.2 to R.sup.7 are as defined in any one of claims 52 and 55 to 59.

63. A compound as claimed in claim 52 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00139##

64. A compound as claimed in claim 52 selected from any of the following compounds, or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00140##

65. A compound of formula (V), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00141## wherein: R.sup.1 is a 5-membered, saturated or unsaturated, carbocyclic ring substituted by one or more substituents, or R.sup.1 is a 6-membered, unsaturated, carbocyclic ring substituted by one or more substituents, or R.sup.1 is a 6-membered, saturated, carbocyclic ring substituted by one or three substituents; R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.7 is H, OH, or OC(O)NR.sub.2 (where each R is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), preferably R.sup.7 is H, OH or OC(O)NH.sub.2; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. C.sub.1-6 alkyl group); and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

66. A compound as claimed in claim 65, wherein R.sup.1 is a cyclohexyl ring substituted by one or three substituents, or a cyclopentyl ring substituted by one or more substituents, for example one or two substituents.

67. A compound as claimed in claim 65, wherein R.sup.1 is a cyclohexenyl ring which is optionally substituted by one or more substituents.

68. A compound as claimed in any one of claims 65 to 67, wherein R.sup.1 is substituted by one or more of the following groups: OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), C.sub.1-3 alkyl (e.g. CH.sub.3), CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) and SO.sub.3H.sub.2 (or an ester thereof).

69. A compound as claimed in claim 65, wherein R.sup.1 is selected from any of the following groups (in which * denotes the point of attachment of the substituent to the remainder of the molecule): ##STR00142##

70. A compound as claimed in any one of claims 65 to 69, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

71. A compound as claimed in claim 70, wherein R.sup.8 is ethyl.

72. A compound as claimed in any one of claims 65 to 71, wherein each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

73. A compound as claimed in claim 65 of formula (Va) or a pharmaceutically acceptable, salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00143## wherein R.sup.1 to R.sup.7 are as defined in any one of claims 65 to 69.

74. A compound as claimed in claim 65 of formula (Vb), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00144## wherein R.sup.1 to R.sup.7 are as defined in any one of claims 65 to 69.

75. A compound as claimed in any one of claims 65 to 74, wherein R.sup.2 is Cl.

76. A compound as claimed in any one of claims 65 to 75, wherein R.sup.3 is OH.

77. A compound as claimed in any one of claims 65 to 76, wherein R.sup.4 and R.sup.5 together are O, or wherein R.sup.4 is H and R.sup.5 is OH.

78. A compound as claimed in any one of claims 65 to 77, wherein R.sup.6 is Cl.

79. A compound as claimed in claim 65 selected from any of the following compounds, their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs: ##STR00145## ##STR00146##

80. A compound as claimed in claim 65 selected from any of the following compounds, their pharmaceutically acceptable salts, metabolites, and prodrugs: ##STR00147## ##STR00148##

81. A compound of formula (VI), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00149## wherein: R.sup.1 is a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or R.sup.1 is an optionally substituted straight-chained or branched C.sub.1-6 alkyl group (e.g. C.sub.1-3 alkyl group); R.sup.2 is H, F, Cl, Br, I or CH.sub.3; R.sup.3 is H or OH; R.sup.4 and R.sup.5 are independently selected from H and OH, or R.sup.4 and R.sup.5 together are O; R.sup.6 is H, F, Cl, Br, I or CH.sub.3; R.sup.8 is a straight-chained or branched C.sub.1-8 alkyl group (e.g. C.sub.1-6 alkyl group); and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds; with the proviso that either at least one of R.sup.2, R.sup.3 and R.sup.6 is H, or R.sup.4 is H and R.sup.5 is OH).

82. A compound as claimed in claim 81, wherein R.sup.1 is an optionally substituted cyclohexyl or cyclopentyl ring, an optionally substituted cyclohexenyl ring, or an optionally substituted straight-chained C.sub.1-6 alkyl group.

83. A compound as claimed in claim 81 or claim 82, wherein R.sup.1 is substituted by one or more of the following groups: OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), C.sub.1-3 alkyl (e.g. CH.sub.3), CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) and SO.sub.3H.sub.2 (or an ester thereof).

84. A compound as claimed in any one of claims 81 to 83, wherein R.sup.8 is a straight-chained or branched C.sub.1-5 alkyl, preferably a straight-chained or branched C.sub.1-4 alkyl, e.g. methyl, ethyl, isopropyl, or tert. butyl.

85. A compound as claimed in claim 84, wherein R.sup.8 is ethyl.

86. A compound as claimed in claim 81 of formula (VIa), or a pharmaceutically acceptable, salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00150## wherein: R.sup.d is H, OH, NR.sup.a.sub.2 (where each R.sup.a is independently H or C.sub.1-3 alkyl, e.g. CH.sub.3), SR.sup.b (where R.sup.b is H or C.sub.1-3 alkyl, e.g. CH.sub.3), halogen (e.g. F, Cl, Br, or I), or C.sub.1-3 alkyl (e.g. CH.sub.3), preferably R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3; R.sup.e is H, CO.sub.2H (or an ester thereof), PO.sub.3H.sub.2 (or an ester thereof) or SO.sub.3H.sub.2 (or an ester thereof), preferably R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2; R.sup.2 to R.sup.6 are as defined in claim 81; and each independently represents an optional bond (i.e. each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds).

87. A compound as claimed in claim 86, wherein: R.sup.d is H, OH, NH.sub.2, SH, F, Cl, Br, I, or CH.sub.3, preferably OH; R.sup.e is H, CO.sub.2H, PO.sub.3H.sub.2, or SO.sub.3H.sub.2, preferably CO.sub.2H; each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9, and C.sub.10-C.sub.11 are independently either CC (single) or CC (double) bonds; R.sup.2 is H, Cl or Br, e.g. R.sup.2 is H or Cl; R.sup.3 is H or OH; R.sup.4 is H and R.sup.5 is OH; and R.sup.6 is H, Cl or Br, e.g. R.sup.6 is H or Cl.

88. A compound as claimed in any one of claims 81 to 87, wherein R.sup.2 is H or Cl, preferably H, or wherein R.sup.2 is Br.

89. A compound as claimed in any one of claims 81 to 88, wherein R.sup.3 is H.

90. A compound as claimed in any one of claims 81 to 89, wherein R.sup.4 is H and R.sup.5 is OH.

91. A compound as claimed in any one of claims 81 to 90, wherein R.sup.6 is H or Cl, preferably H, or wherein R.sup.6 is Br.

92. A compound as claimed in any one of claims 81 to 91, wherein each of C.sub.2-C.sub.3, C.sub.4-C.sub.5, C.sub.6-C.sub.7, C.sub.8-C.sub.9 and C.sub.10-C.sub.11 are CC (double) bonds.

93. A compound as claimed in claim 81 of formula (VIb), or a pharmaceutically acceptable salt, metabolite, isomer (e.g. stereoisomer) or prodrug thereof: ##STR00151## wherein R.sup.d, R.sup.e, and R.sup.2 to R.sup.6 are as defined in any one of claims 81 and 86 to 91.

94. A compound as claimed in claim 81 of formula (VIc), or a pharmaceutically acceptable salt, metabolite, or prodrug thereof: ##STR00152## wherein R.sup.d, R.sup.e, and R.sup.2 to R.sup.6 are as defined in any one of claims 81 and 86 to 91.

95. A compound as claimed in claim 81 selected from any one of the following compounds, their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs: ##STR00153## ##STR00154##

96. A compound as claimed in claim 81 selected from any one of the following compounds, their pharmaceutically acceptable salts, metabolites, and prodrugs: ##STR00155## ##STR00156##

97. A compound as claimed in any one of claims 1 to 96 for use as a medicament.

98. A compound as claimed in any one of claims 1 to 96 for use as an antimicrobial agent.

99. A compound as claimed in any one of claims 1 to 96 for use in the treatment of an infection caused by a microbe which is a bacterium.

100. A compound for use as claimed in claim 99 in the treatment of an infection caused by a microbe which is a Gram-negative bacterium, e.g. selected from Acinetobacter species, Burkholderia species, Ralstonia species and Stenotrophomonas species.

101. A compound as claimed in any one of claims 1 to 96 for use in the treatment of an infection caused by at least one microbe which is resistant to at least one antimicrobial drug.

102. A compound for use as claimed in claim 101 in the treatment of an infection, wherein the antimicrobial drug is selected from drugs of the carbapenem family, drugs of the penicillin family, drugs of the vancomycin family, drugs of the aminoglycoside family, drugs of the quinolone family, drugs of the daptomycin family, drugs of the cephalosporin family, drugs of the macrolide family, and combinations thereof.

103. A compound for use as claimed in claim 102 in the treatment of an infection, wherein the antimicrobial drug is selected from penicillin, ampicillin, methicillin, vancomycin, gentamycin, ofloxacin, ciprofloxacin, daptomycin, cefdimir, erythromycin, equivalents thereof, and combinations thereof.

104. Use of a compound as claimed in any one of claims 1 to 96 in the manufacture of a medicament for use in treating an infection caused by at least one microbe as defined in any one of claims 99 to 103.

105. A compound for use as claimed in any one of claims 99 to 103 in the treatment of infection, or a use as claimed in claim 104, wherein the infection is an infection of the respiratory system, digestive system, urinary system, nervous system, a blood infection, a soft tissue infection, a skin infection, a nasal canal infection, or combinations thereof.

106. A pharmaceutical composition comprising a compound as claimed in any one of claims 1 to 96 and a pharmaceutically acceptable carrier.

107. A pharmaceutical composition as claimed in claim 106, further comprising at least one other therapeutically active agent.

108. A pharmaceutical composition as claimed in claim 107, wherein the compound according to any one of claims 1 to 96 and the other therapeutically active agent are adapted for sequential, separate or simultaneous administration.

109. An active agent, especially an antimicrobial agent, having mass spectral and/or NMR spectroscopic properties substantially according to one or more of FIGS. 1 to 126 and/or any one of Tables 3 to 27.

110. A process for the preparation of a compound as claimed in any one of claims 1 to 96, comprising cultivating a microorganism capable of producing said compound, in a culture medium comprising a source of assimilable carbon, nitrogen, and inorganic salts and, optionally, recovering said compound from the culture medium and, optionally, further converting the compound into a pharmaceutically acceptable salt thereof.

111. A process as claimed in claim 110, wherein the microorganism is Burkholderia ambifaria, e.g. one or more strains selected from BCCC0203 (also known as LMG P-24640; BCF), BCC0118 (also known as LMG P-24636; JLO), BCC 1248 (also known as LMG P-24641; KWO-1), BCC0250 (also known as LMG P-24637; WM2), BCC1241 (also known as LMG P-24639; KC311-6), BCC0207 (also known as LMG P-19182; AMMD; ATCC BAA-244; CCUG 44356; KCTC 12943; FC768; J2742 Vandamme R-696FC0768), and BCC0267 (also known as LMG P-19467; CEP0996; Coenye R-9935), or a mutant or variant thereof.

112. A process as claimed in claim 110 or claim 111, further comprising converting the compound into another compound of any one of formula (I) to (VI) by chemical synthesis and, optionally, further converting the resultant compound into a pharmaceutically acceptable salt thereof.

113. A method for the treatment of an infection, the method comprising administering to a subject in need thereof a compound as claimed in any one of claims 1 to 96, wherein the infection is caused by at least one microbe, optionally wherein the microbe is resistant to an antimicrobial drug.

Description

[0439] The invention will now be further illustrated by the following non-limiting examples and the accompanying figures, in which:

[0440] FIGS. 1 to 126 show .sup.1H NMR, .sup.13C NMR, COSY NMR, .sup.1H-.sup.13C HSQC and .sup.1H-.sup.13C HMBC spectra of compounds according to the invention.

EXAMPLES

Example 1Production and Purification of Enacyloxin Derivatives

Gene Deletion

[0441] In-frame deletions in genes were introduced via double homologous recombination using the suicide plasmid pGPI and the I-SceI expression plasmid pDAI (Flannagan et al., Environ. Microbiol. 10: 1652-1660, 2008). The sequences (500-1000 bp) flanking the gene regions targeted for deletion were amplified from B. ambifaria BCC0203 genomic DNA using Q5 DNA polymerase (NEB). Restriction sites were introduced at the 5-end of the primers to allow for directional cloning of the PCR products into pGPI. Constructs were mobilized into E. coli SY327 by electroporation and transformants were selected on LB agar plates supplemented with trimethoprim (50 g/mL). Plasmids were purified from trimethoprim-resistant colonies using the GeneJET Plasmid Miniprep kit (Thermo Scientific) and correct assembly of the mutagenesis constructs was confirmed by Sanger sequencing (GATC Biotech). Validated constructs were transferred into B. ambifaria BCC0203 via triparental mating (Agnoli et al., Mol. Microbiol. 83: 362-378, 2012) and transconjugants were selected using trimethoprim (200 g/ml) and gentamicin (50 g/ml). Single B. ambifaria mutants were selected and correct integration of the mutagenesis plasmids into the genome was confirmed by colony PCR. Next, the pDAI plasmid was introduced into the B. ambifaria single crossover mutants by triparental mating using E. coli SY327 (pDAI) and E. coli HB101 (pRK2013) as the donor and helper strain, respectively (Agnoli et al., Mol. Microbiol. 83: 362-378, 2012). Transconjugants were selected on LB agar plates containing tetracycline (200 g/ml) and gentamicin (50 g/ml). Single B. ambifaria mutants were selected and correct gene deletion was confirmed by colony PCR and Sanger sequencing. To examine the effect of the gene deletions on enacyloxin biosynthesis, mutant strains were grown at 30 C. on solid minimal medium containing glycerol as a sole carbon source (BSM-G) (O'Sullivan et al., Environ. Microbiol. 9: 1017-1034, 2007). Following incubation for 3 days, the cells were scraped off and ethyl acetate extracts of the agar were analysed by UHPLC-ESI-Q-TOF-MS.

Production and Purification of the Enacyloxin Derivatives

[0442] For production of enacyloxin derivatives, the B. ambifaria BCC0203 deletion mutants were grown in the dark at 30 C. on solid minimal medium containing glycerol as a sole carbon source (BSM-G) (O'Sullivan et al., Environ. Microbiol. 9: 1017-1034, 2007). Following incubation for 3 days, the cells were scraped off and the agar extracted twice using ethyl acetate (1:1). Enacyloxin derivative-containing extracts were concentrated by rotary evaporation in vacuo and the resulting residue was re-dissolved in acetonitrile for purification by preparative HPLC. For the mutasynthetic production of enacyloxin derivatives, B. ambifaria BCC0203 mutant 5912-5914 was grown in the dark at 30 C. on BSM-G agar supplemented with 10 mM of a relevant DHCCA analogue (described below). Bromine derivatives of enacyloxin were generated using the same procedure, however the ammonium chloride in the BSM-G was replaced with two equivalents of ammonium bromide.

[0443] Enacyloxin derivative-containing ethyl acetate extracts of the agar were fractionated by preparative HPLC using an Agilent 1260 instrument equipped with a Zorbax SB-C.sub.18 column (21.2100 mm, 5 m), monitoring absorbance at 360 nm. Mobile phases consisted of water and acetonitrile, each supplemented with 0.1% formic acid. A gradient of 50% B to 100% B over 40 minutes was employed at a flow rate of 10 mL/min. Enacyloxin derivative-containing fractions were pooled, concentrated in vacuo, and subsequently lyophilized. To avoid photo-induced isomerization of the compounds, exposure to light was minimized throughout the entire purification process.

Synthesis of DHCCA DerivativesGeneral Procedures

[0444] Room temperature refers to ambient temperature (20-22 C.), 5 C. refers to a cold water bath and 00 C. refers to an ice slush bath. Heated experiments were conducted using thermostatically controlled oil baths. All commercially available solvents and chemicals were used without any further purification. NMR spectra were recorded on Bruker Advance AV-300 and HD-500 MHz spectrometers at room temperature (298 K). Chemical shifts are reported in parts per million (ppm) referenced from CDCl3 (H: 7.26 ppm and C: 77.0 ppm). Coupling constants (J) are rounded to the nearest 0.5 Hertz (Hz). Multiplicities are given as multiplet (m), singlet (s), doublet (d), triplet (t), quartet (q), quintet (quin.), sextet (sext.), septet (sept.), octet (oct.) and nonet (non.). 1H and 13C assignments were established on the basis of COSY, DEPT, HMQC and HMBC correlations. Infra-red spectra were recorded using either a Perkin Elmer Spectrum 100 FT-IR spectrometer or an Alpha Bruker Platunium ATR single reflection diamond ATR module. Optical rotations were measured using an Optical Activity Ltd AA-1000 millidegree auto-ranging polarimeter (589 nm). Specific rotations are given in units of 10-1 deg cm.sup.2g.sup.1. Melting points were recorded on a Stuart scientific melting point apparatus and are uncorrected. Silica column chromatography was performed on 40-60 silica gel. Thin layer chromatography (TLC) was carried out aluminum sheets coated with 0.2 mm silica gel 60 F254. Visualisation was effected by UV light (254 nm) or by potassium permanganate solution followed by heating. Low resolution mass spectra (LRMS) were recorded using an Agilent 6130B single Quad (ESI). High resolution mass spectra (HRMS) were obtained using a Bruker micro-TOF ESI spectrometer.

Synthesis of Cyclopentane DHCCA Derivative(1S,3R,4S)-3,4-dihydroxycyclopentane-1-carboxylic acid (2)

[0445] ##STR00047##

[0446] Acetonide (1) was synthesized according to literature procedure (WO 2013/170030). To a solution of acetonide (1) (328 mg, 1.64 mmol) in MeOH (10 mL) was added TsOH (35 mg, 0.20 mmol) and the reaction was stirred at room temperature for 1 hour. The mixture was then concentrated in vacuo and partitioned between EtOAc (20 mL) and saturated NaHCO.sub.3 (20 mL). The layers were separated and the aqueous phase further extracted with EtOAc (220 mL), the combined organics were then washed with brine, dried (MgSO.sub.4), filtered and concentrated in vacuo to afford the crude diol as a viscous oil. The diol was then dissolved in THF (10 mL) and H.sub.2O (5 mL), and LiOH was added (40 mg, 2 mmol). The reaction was then stirred at room temperature overnight and concentrated in vacuo to afford the product as a viscous oil (162 mg, 78%).

[0447] .sub.H (500 MHz, MeOD) 4.09-4.02 (2H, br. m, CHOH), 3.06 (1H, tt, J 9.5 and 3, CHCOOH), 2.09-1.88 (4H, m, CH.sub.2); .sub.C (125 MHz, MeOD) 183.2 (CO.sub.2H), 73.2 (CHOH), 40.1 (CHCOOH), 35.1 (CH.sub.2); HRMS (ESI) cald. for C.sub.6H.sub.10NaO.sub.4 (M+Na.sup.+) requires 169.0477, found 169.0475.

Synthesis of (1R,3R,4R)-3,4-dihydroxycyclohexane-1-carboxylic acid (3)

[0448] ##STR00048##

[0449] (1R,3R,4R)-3,4-dihydroxycyclohexane-1-carboxylic acid (3) was synthesized and used as a racemic mixture by following literature procedure (WO 2015/4455).

Synthesis of Unsaturated DHCCA Derivative(4S,5R)-4,5-dihydroxycyclohex-1-ene-1-carboxylic acid (7)

[0450] ##STR00049##

[0451] Alcohol (4) was synthesized according to literature procedure (Z. Wang et al., J. Org. Chem., 1997, 62, 8622-8623). To a stirred solution of alcohol (4) (370 mg, 1.62 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added Et.sub.3N (2.25 mL, 16.2 mmol) and methanesulfonyl chloride (0.63 mL, 8.11 mmol) at 000 C. and the mixture was stirred overnight at room temperature. The reaction was quenched with 1M HCl (10 mL) and the mixture extracted with CH.sub.2Cl.sub.2 (310 mL), the combined organics were then washed with brine, dried (MgSO.sub.4), filtered and concentrated in vacuo to afford the crude product as an orange solid. The crude product was purified by silica chromatography (EtOAc:Petroleum ether, 60:40) to afford methyl (3aR,7aR)-2,2-dimethyl-7-((methylsulfonyl)oxy)-3a,4,7,7a-tetrahydrobenzo[d][1,3]dioxole-5-carboxylate (5) as a white solid (411 mg, 83%).

[0452] .sub.H (500 MHz, CDCl.sub.3) 6.95 (1H, br. s, CHCHOS), 5.06 (1H, br. s, CHOS), 4.75-4.71 (1H, m, CH.sub.2CHOCHO), 4.67-4.63 (1H, m, CH.sub.2CHO), 3.78 (3H, s, OCH.sub.3), 3.17 (3H, s SCH.sub.3), 3.07 (1H, d, J 16.5, CH.sub.2), 2.00 (1H, d, J 16.5, CH.sub.2), 1.34 (3H, s, CCH.sub.3), 1.32 (3H, s, CCH.sub.3); .sub.C (125 MHz, CDCl.sub.3) 165.5 (CO.sub.2Me), 136.2 (CHCOS), 130.8 (CCOOMe), 109.9 (C(CH.sub.3).sub.2), 75.7 (CHOS), 75.4 (CH.sub.2CHOCHO), 72.5 (CH.sub.2CHO), 52.4 (COOCH.sub.3), 39.2 (SCH.sub.3), 27.2 (CH.sub.2), 25.8 (CH.sub.3), 24.4 (CH.sub.3); HRMS (ESI) cald. for C.sub.12H.sub.18NaO.sub.7S (M+Na.sup.+) requires 329.0671, found 329.0672.

[0453] To a stirred solution of Pd.sub.2(dba).sub.3.CHCl.sub.3 (60 mg, 0.06 mmol), tributylphosphine (15 l. 0.06 mmol) and mesylate (5) (330 mg, 1.1 mmol) in dioxane (10 mL) under and argon atmosphere was added a suspension of NaBH.sub.4 (40 mg, 1.1 mmol) in H.sub.2O (1 mL) and the reaction was stirred for 2 hours at room temperature. The reaction mixture was then diluted with saturated brine (10 mL) and extracted with Et.sub.2O (320 mL), the combined organics were then washed with brine, dried (MgSO.sub.4), filtered and concentrated in vacuo to afford the crude product as a black oil. The crude product was purified by silica chromatography (EtOAc:Petroleum ether, 10:90) to afford methyl (3aR,7aS)-2,2-dimethyl-3a,4,7,7a-tetrahydrobenzo[d][1,3]dioxole-5-carboxylate (6) as a colourless oil (159 mg, 68%).

[0454] .sub.H (500 MHz, MeOD) 7.02 (1H, dt, J 6 and 3, CHCH.sub.2CHO), 4.52 (1H, ddd, J 7, 4 and 3, CH.sub.2CHO), 4.48 (1H, ddd, J 7, 5 and 2.5, CHCH.sub.2CHO), 3.74 (3H, s, OCH.sub.3), 2.74 (1H, dd, J 16.5 and 3, CH.sub.2C), 2.50 (1H, dd, J 17, 6 and 2, CHCH.sub.2), 2.29-2.20 (2H, m, CH.sub.2C, CHCH.sub.2), 1.29 (3H, s, CCH.sub.3), 1.28 (3H, s, CCH.sub.3); .sub.C (125 MHz, MeOD) 168.4 (CO.sub.2Me), 139.5 (CHCH.sub.2), 130.7 (CCOOMe), 108.9 (C(CH.sub.3).sub.2), 74.7 (CH.sub.2CHO), 74.0 (CHCH.sub.2CHO), 52.2 (COOCH.sub.3), 30.0 (CH.sub.2C), 28.0 (CHCH.sub.2) 26.7 (CH.sub.3), 24.6 (CH.sub.3); HRMS (ESI) cald. for C.sub.11H.sub.16NaO.sub.4 (M+Na.sup.+) requires 235.0946, found 235.0949.

[0455] To a solution of acetonide (6) (150 mg, 0.7 mmol) in MeOH (10 mL) was added TsOH (17 mg, 0.1 mmol) and the reaction was stirred at room temperature for 1 hour. The mixture was then concentrated in vacuo and partitioned between EtOAc (20 mL) and saturated NaHCO.sub.3 (20 mL). The layers were separated and the aqueous phase further extracted with EtOAc (220 mL), the combined organics were then washed with brine, dried (MgSO.sub.4), filtered and concentrated in vacuo to afford the crude diol as a viscous oil. The diol was then dissolved in THF (10 mL) and H.sub.2O (5 mL), and LiOH was added (20 mg, 1 mmol). The reaction was then stirred at room temperature overnight and concentrated in vacuo to afford (4S,5R)-4,5-dihydroxycyclohex-1-ene-1-carboxylic acid (7) as a viscous oil (81 mg, 73%).

[0456] .sub.H (500 MHz, MeOD) 6.87-6.84 (1H, m, OHCH.sub.2CHO), 3.92-3.84 (2H, m, CHO), 2.55-2.35 (4H, m, CH.sub.2), .sub.C (125 MHz, MeOD) 170.25 (CO.sub.2H), 137.8 (CHCH.sub.2), 128.9 (CCOOH), 69.6 (CH.sub.2CHO), 68.5 (CHCH.sub.2CHO), 32.3 (CH.sub.2C), 31.0 (CHCH.sub.2) 26.7 (CH.sub.3); HRMS (ESI) cald. for C.sub.7H.sub.10NaO.sub.4 (M+Na.sup.+) requires 181.0477, found 181.0476.

Structure Elucidation of the Enacyloxin Derivatives

[0457] Structure elucidation of the compounds was achieved using a combination of UHPLC-ESI-Q-TOF-MSIMS and 1- and 2-D NMR experiments. UHPLC-ESI-Q-TOF-MS analyses were performed on a Zorbax Eclipse Plus C.sub.18 column (1.8 m, 2.1100 mm, Agilent) coupled to a Bruker MaXis Impact mass spectrometer. Mobile phases consisted of water and acetonitrile (ACN), each supplemented with 0.1% formic acid. A gradient of 30% ACN to 100% ACN over 34 minutes was employed at a flow rate of 0.2 mL/min. The mass spectrometer was operated in positive ion mode with a scan range of 50-3000 m/z. UV absorbance was monitored at 360 nm.

[0458] For NMR analysis, purified compounds were dissolved in d4-MeOH and .sup.1H, .sup.13C, COSY, HSQC and HMBC spectra were recorded on a Bruker Avance 500 MHz spectrometer equipped with a DCH cryoprobe at 25 C.

[0459] Knock-out mutagenesis or mutasynthesis methods as described above were used to prepare the compounds in Table 2.

TABLE-US-00002 TABLE 2 Polyketide compounds of the invention Compound No. Gene manipulation method Structure of compound 1 B. ambifaria BCCO203 bamb_5927 [00050] 2 B. ambifaria BCCO203 bamb_5931 [00051] 3 B. ambifaria BCCO203 bamb_5930 [00052] 4 B. ambifaria BCCO203 bamb_5912 [00053] 5 B. ambifaria BCCO203 bamb_5927, bamb_5930 [00054] 6 B. ambifaria BCCO203 bamb_5927, bamb_5931 [00055] 7 B. ambifaria BCCO203 bamb_5928 [00056] 8 B. ambifaria BCCO203 bamb_5932 [00057] 9 B. ambifaria BCCO203 bamb_5930, bamb_5932 [00058] 10 B. ambifaria BCCO203 bamb_5930, bamb_5931 [00059] 11 B. ambifaria BCCO203 bamb_5930-5932 [00060] 12 B. ambifaria BCCO203 bamb_5927-5932 [00061] 13 B. ambifaria BCC0203 5912-5914 + 3-amino-4- hydroxycyclohexane carboxylic acid [00062] 14 B. ambifaria BCC0203 5912-5914 + 3-amino-4- hydroxycyclohexane carboxylic acid [00063] 15 B. ambifaria BCC0203 5912-5914 + 3-amino-4- hydroxycyclohexane carboxylic acid [00064] 16 B. ambifaria BCC0203 5912-5914 + 3,4- dihydroxycyclohexane carboxylic acid [00065] 17 B. ambifaria BCC0203 5912-5914 + 3,4- dihydroxycyclohexane carboxylic acid [00066] 18 B. ambifaria BCC0203 5912-5914 + 3- hydroxycyclopentane carboxylic acid [00067] 19 B. ambifaria BCC0203 5912-5914 + 3,4- diaminocyclohexane carboxylic acid [00068] 20 B. ambifaria BCC0203 5912-5914 + syn-3- aminocyclopentane carboxylic acid [00069] 21 B. ambifaria BCC0203 5912-5914 + 3- aminocyclohexane carboxylic acid [00070] 22 B. ambifaria BCC0203 5912-5914 + 3- aminocyclohexane carboxylic acid [00071] 23 B. ambifaria BCC0203 5912-5914 + 4-amino-3- hydroxybutyric acid [00072] 24 B. ambifaria BCC0203 5912-5914 + 4- aminobutyric acid [00073] 25 B. ambifaria BCC0203 5912-5914 + shikimate [00074] 26 B. ambifaria BCC0203 5912-5914 + shikimate [00075] 27 B. ambifaria BCC0203 5912-5914 + shikimate [00076] 28 B. ambifaria BCC0203 5912-5914 + DHCCA derivative (7) [00077] 29 B. ambifaria BCC0203 5912-5914 + 5930 + 5932 + DHCCA derivative (7) [00078] 30 B. ambifaria BCC0203 [00079] 31 B. ambifaria BCC0203 [00080] 32 B. ambifaria BCC0203 [00081] 33 B. ambifaria BCC0203 bamb_5930, bamb_5932 [00082] 34 B. ambifaria BCC0203 bamb_5930, bamb_5932 [00083] 35 B. ambifaria BCC0203 bamb_5930, bamb_5932 [00084] 36 B. ambifaria BCC0203 bamb_5927 [00085] 37 B. ambifaria BCC0203 bamb_5927 [00086] 38 B. ambifaria BCC0203 bamb_5927 [00087] 39 B. ambifaria BCC0203 bamb_5930 [00088] 40 B. ambifaria BCC0203 bamb_5930 [00089] 41 B. ambifaria BCC0203 bamb_5930 [00090]

[0460] Structures were confirmed by .sup.1H NMR, .sup.13C NMR, COSY NMR, .sup.1H-.sup.13C HSQC and .sup.1H-.sup.13C HMBC and spectra are shown in FIGS. 1 to 61. The resulting chemical shift assignments for the compounds are listed in Tables 3 to 27.

TABLE-US-00003 TABLE 3 NMR assignments for 18-dechloro-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00091] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.53 (m) 39.6 2 1.73/2.17 (m/dm, 14.2) 32.8 3 5.19 (m) 73.8 4 3.72 (m) 70.7 5 1.80 (m) 29.6 6 1.56/2.02 (m/m) 28.1 1 168.6 2 6.01 (d, 15.0) 121.7 3 7.43 (dd, 15.0, 11.0) 147.0 4 6.52 (dd, 15.0, 11.0) 127.3 5 6.76 (d, 15.0) 147.0 6 137.1 6-Me 1.95 (s) 12.9 7 6.41 (br, d, 10.0) 137.4 8 6.78 (dd, 14.9, 10.0) 132.1 9 6.71 (dd, 14.9, 9.9) 131.9 10 6.45 (d, 9.8) 128.6 11 140.8 12 2.93 (dq, 9.5, 6.7) 47.7 12-Me 1.18 (d, 6.5) 13.1 13 4.05 (dd, 9.5, 1.5) 74.3 14 4.22 (d, 1.7) 79.2 15 211.1 16 2.64/2.82 (dd, 16.0, 47.3 4.0/dd, 16.0, 8.0) 17 4.19 (m) 65.3 18 1.80/1.64 (ddd, 14.5, 10, 43.9 3/ddd, 14.5, 9.5, 3.5) 19 5.22 (ddd, 9.5, 7, 3.5) 73.0 19-carbamate 158.0 20 5.43 (ddt, 15.5, 7.0, 1.5) 126.5 21 5.76 (dt, 15.5, 6.5) 139.7 22 2.03 (qdd, 7.9, 6.4, 1.5) 26.8 23 1.00 (t, 7.4) 14.0

TABLE-US-00004 TABLE 4 NMR assignments for 14-dehydroxy-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00092] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.53 (m) 39.8 2 1.73/2.17 (m/dm, 14.2) 32.9 3 5.21 (m) 73.5 4 3.73 (m) 70.8 5 1.81 (m) 29.5 6 1.56/2.02 (m/m) 28.3 1 168.5 2 6.03 (d, 15.0) 121.6 3 7.44 (dd, 15.0, 11.0) 147.0 4 6.54 (dd, 15.0, 11.0) 127.4 5 6.76 (d, 15.0) 146.8 6 136.6 6-Me 1.97 (br, s) 12.9 7 6.42 (br, d, 10.0) 137.0 8 6.77 (dd, 14.8, 10.0) 131.8 9 6.71 (dd, 14.8, 9.9) 131.5 10 6.45 (d, 9.9) 128.4 11 140.2 12 2.66 (dq, 9.6, 6.7) 48.5 12-Me 1.16 (d, 7.0) 13.1 13 4.24 (ddd, 9, 7.5, 3.5) 67.8 14 2.61 (d, 9) 49.5 15 209.6 16 2.78/2.90 (dd, 17.0, 5.0/dd, 44.9 17.0, 8.0) 17 4.46 (m) 67.3 18 4.00 (dd, 8.5, 2.5) 68.0 19 5.26 (t, 7.5) 75.8 19-carbamate 158.1 20 5.54 (ddt, 15.4, 7.4, 1.4) 126.3 21 5.89 (dt, 15.2, 6.4) 139.7 22 2.10 (qdd, 7.9, 6.4, 1.4) 26.8 23 1.01 (t, 7.4) 14.0

TABLE-US-00005 TABLE 5 NMR assignments for decarbamoyl-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00093] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.2 1 2.54 (m) 39.1 2 1.72/2.18 (m/dm, 14.2) 32.8 3 5.20 (m) 73.5 4 3.73 (m) 70.6 5 1.81 (m) 29.6 6 1.56/2.01 (m/m) 28.1 1 168.5 2 6.03 (d, 15.0) 121.6 3 7.43 (dd, 15.0, 11.0) 146.8 4 6.54 (dd, 15.0, 11.0) 127.4 5 6.75 (d, 15.0) 146.6 6 137.0 6-Me 1.96 (br, s) 12.6 7 6.41 (br, d, 10.0) 137.0 8 6.77 (dd, 14.9, 10.0) 131.8 9 6.73 (dd, 14.8, 9.9) 131.5 10 6.43 (d, 9.9) 128.4 11 141.0 12 2.90 (dq, 9.6, 6.7) 47.9 12-Me 1.12 (d, 6.5) 16.0 13 4.17 (dd, 9.5, 0.5) 72.7 14 3.45 (d, 0.5) 73.2 15 100.5 16 1.53/2.45 (dd, 13, 12/dd, 42.2 13, 5.0) 17 3.97 (ddd, 11.5, 9.5, 5) 70.8 18 3.36 (t, 9.5) 67.5 19 5.26 (dd, 7.8, 7.6) 74.9 20 5.54 (ddt, 15.5, 7, 1.5) 127.2 21 5.84 (dt, 15.2, 6.4) 137.7 22 2.10 (qdd, 7.9, 6.4, 1.5) 26.7 23 1.01 (t, 7.4) 13.8

TABLE-US-00006 TABLE 6 NMR assignments for 5-hydroxy-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00094] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 178.5 1 2.66 (m) 38.8 2 1.70/2.09 (m/dm, 14.2) 32.9 3 5.30 (q, 3) 73.7 4 3.45 (dd, 9.5, 3) 74.4 5 3.82 (ddd, 11.5, 9.5, 5) 69.9 6 1.48/2.24 (m/m) 35.7 1 168.4 2 6.00 (d, 15.0) 121.2 3 7.40 (dd, 15.0, 11.0) 147.0 4 6.52 (dd, 15.0, 11.0) 127.4 5 6.74 (d, 15.0) 146.6 6 137.5 6-Me 1.95 (br, s) 12.6 7 6.41 (br, d, 10.0) 137.5 8 6.75 (dd, 14.8, 10.0) 132.2 9 6.72 (dd, 14.8, 9.9) 131.5 10 6.43 (d, 9.9) 128.4 11 140.7 12 2.93 (dq, 9.6, 6.7) 47.6 12-Me 1.19 (d, 6.5) 16.3 13 4.04 (br, d, 9.4) 74.2 14 4.22 (d, 1.7) 79.0 15 211.5 16 2.82/3.03 (dd, 17.0, 4.5/dd, 44.6 16.9, 8.0) 17 4.50 (m) 66.8 18 4.04 (m) 68.0 19 5.26 (dd, 7.8, 7.6) 75.9 19-carbamate 158.7 20 5.54 (ddt, 15.4, 7.4, 1.4) 126.2 21 5.88 (dt, 15.2, 6.4) 139.2 22 2.09 (qdd, 7.9, 6.4, 1.4) 26.4 23 1.01 (t, 7.4) 13.7

TABLE-US-00007 TABLE 7 NMR assignments for 14-dehydroxy-18-decarbamoyl-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00095] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.55 (m) 40.1 2 1.80/2.19 (m/dm, 14.2) 32.6 3 5.21 (m) 73.2 4 3.73 (m) 70.6 5 1.83 (m) 29.5 6 1.55/2.01 (m/m) 27.8 1 168.5 2 6.03 (d, 15.0) 121.5 3 7.43 (dd, 15.0, 11.0) 146.8 4 6.54 (dd, 15.0, 11.0) 127.3 5 6.75 (d, 15.0) 146.6 6 137.0 6-Me 1.97 (br, s) 12.6 7 6.41 (br, d, 10.0) 137.5 8 6.78 (dd, 14.8, 10.0) 131.6 9 6.72 (dd, 14.8, 9.9) 130.1 10 6.43 (d, 10.0) 128.2 11 140.1 12 2.58 (m) 51.2 12-Me 1.14 (d, 6.5) 15.3 13 4.25 (dt, 9, 1.5) 70.5 14 1.7 (d, 10.5) 32.6 15 98.9 16 1.55/2.21 (dd, 12.5, 45.3 11.5/dd, 7.5, 5) 17 3.97 (ddd, 11.5, 9.5, 5) 70.7 18 3.34 (d, 10) 67.5 19 4.31 (dd, 10, 6.5) 75.9 20 5.52 (ddt, 15.5, 8.5, 1.5) 127.3 21 5.84 (dt, 15.5, 6.5) 136.9 22 2.09 (qdd, 7.9, 6.4, 1.4) 26.4 23 1.02 (t, 7.4) 13.8

TABLE-US-00008 TABLE 8 NMR assignments for 14-dehydroxy-18-dechloro-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00096] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.53 (m) 39.1 2 1.74/2.17 (m/dm, 14.5) 32.4 3 5.20 (m) 73.3 4 3.75 (m) 70.5 5 1.83 (m) 29.3 6 1.43/2.02 (m/m) 27.8 1 168.9 2 6.04 (d, 15.5) 121.4 3 7.44 (dd, 15.0, 11.0) 147.1 4 6.55 (dd, 15.0, 11.0) 127.1 5 6.75 (d, 15.0) 146.8 6 137.1 6-Me 1.96 (s) 12.7 7 6.41 (br, d, 10.0) 137.4 8 6.78 (dd, 14.9, 10.0) 132.7 9 6.71 (dd, 14.9, 9.9) 131.4 10 6.42 (d, 9.8) 127.6 11 141.0 12 2.59 (m) 51.0 12-Me 1.14 (d, 6.5) 13.9 13 3.98 (m) 70.2 14 2.59 (m) 51.0 15 210.9 16 1.64/1.54 (m/m) 43.3 17 4.10 (m) 65.9 18 1.74/ 1.64 (m, m) 43.9 19 5.24 (ddd, 9.5, 7, 3.5) 73.3 19-carbamate 159.9 20 5.46 (ddt, 15.5, 7.0, 1.5) 129.2 21 5.74 (dt, 15.5, 6.5) 135.8 22 2.05 (qdd, 7.9, 6.4, 1.5) 26.2 23 0.99 (t, 7.4) 13.8

TABLE-US-00009 TABLE 9 NMR assignments for 2-dehydroxy-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00097] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.68 (m) 39.9 2 1.79/2.03 (m/dm, 12.5) 33.8 3 5.15 (m) 70.8 4 1.63 (m) 30.8 5 1.93 (m) 29.4 6 1.57 (m) 21.5 1 168.4 2 5.98 (d, 15.0) 121.4 3 7.39 (dd, 15.0, 11.0) 146.7 4 6.53 (dd, 15.0, 11.0) 127.2 5 6.77 (d, 15.0) 147.6 6 137.0 6-Me 1.96 (ar, s) 12.6 7 6.42 (br, d, 10.0) 137.4 8 6.77 (dd, 14.8, 10.0) 131.6 9 6.72 (dd, 14.8, 9.9) 131.5 10 6.44 (d, 9.9) 128.4 11 140.7 12 2.94 (dq, 9.6, 6.7) 47.6 12-Me 1.19 (d, 6.5) 16.25 13 4.05 (br, d, 9.4) 74.0 14 4.25 (d, 1.7) 78.9 15 211.5 16 2.84/3.05 (dd, 16.9, 4.5/dd, 44.6 16.9, 8.0) 17 4.51 (m) 66.8 18 4.05 (m) 68.0 19 5.28 (dd, 7.8, 7.6) 75.5 19-carbamate 158.7 20 5.56 (ddt, 15.4, 7.4, 1.4) 126.2 21 5.90 (dt, 15.2, 6.4) 139.2 22 2.10 (qdd, 7.9, 6.4, 1.4) 26.3 23 1.02 (t, 7.4) 13.6

TABLE-US-00010 TABLE 10 NMR assignments for enacyloxin-4-amino-3-hydroxybutyric acid conjugate (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00098] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 181.5 1 2.48, 2.39 (m) 41.9 2 4.11 (m) 69.5 3 3.40 (m) 46.9 1 169.5 2 6.10 (d, 15.0) 124.2 3 7.26 (dd, 14.5, 11.0) 145.0 4 6.48 (dd, 15.0, 11.0) 127.5 5 6.76 (d, 15.0) 142.5 6 136.2 6-Me 1.95 (br, s) 12.7 7 6.42 (br, d, 10.0) 137.5 8 6.75 (dd, 14.8, 10.0) 131.7 9 6.69 (dd, 14.8, 9.9) 131.1 10 6.45 (d, 9.9) 128.5 11 139.2 12 2.94 (dq, 9.6, 6.7) 48.2 12-Me 1.19 (d, 6.5) 16.3 13 4.04 (br, d, 9.4) 74.1 14 4.24 (d, 1.7) 78.9 15 211.5 16 2.84/3.05 (dd, 16.9, 4.5/dd, 44.6 16.9, 8.0) 17 4.52 (m) 66.8 18 4.05 (m) 67.9 19 5.28 (dd, 7.8, 7.6) 75.5 19-carbamate 158.7 20 5.55 (ddt, 15.4, 7.4, 1.4) 126.2 21 5.90 (dt, 15.2, 6.4) 140.4 22 2.10 (qdd, 7.9, 6.4, 1.4) 26.3 23 1.02 (t, 7.4) 13.6

TABLE-US-00011 TABLE 11 NMR assignments for 11-dechloro-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00099] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.4 1 2.51 (br. t, 11.1) 39.7 2 1.72/2.19-2.13 (br. t 32.7 11.9/m) 3 5.22-5.18 (m) 73.3 4 3.75-3.69 (m) 70.7 5 1.84-1.77 (2H) (m) 29.6 6 1.60-51/2.05-1.99 (m/m) 27.9 1 168.7 2 5.99 (d, 15.3) 120.9 3 7.42 (dd, 15.0, 11.2) 147.0 4 6.48 (dd, 15.2, 10.8) 128.6 5 6.73 (d, 15.2) 146.9 6 135.7 6-Me 1.94 (s) 12.6 7 6.35 (d, 11.0) 137.6 8 6.41 (dd, 14.8, 10.5) 137.4 9 6.48 (dd, 11.3, 15.1) 126.4 10 6.29 (dd, 15.0, 10.8) 132.8 11 5.86 (dd, 15.0, 8.4) 140.2 12 2.60 (tq, 8.5, 7.4) 41.4 12-Me 1.10 (d, 6.8) 17.3 13 3.73 (dd, 8.4, 2.3) 76.7 14 4.22 (d, 2.6) 79.4 15 211.7 16 3.02/2.83 (dd 17.1, 8.0/dd, 44.7 17.2, 4.8) 17 4.49 (ddd, 7.5, 4.5, 2.4) 66.7 18 4.02 (dd, 8.0, 2.5) 68.0 19 5.26 (t, 7.8) 75.5 19-carbamate 158.6 20 5.54 (ddt, 15.4, 7.4, 1.5) 126.1 21 5.89 (dt, 15.4, 6.5) 139.2 22 2.09 (qdd, 7.6, 6.3, 1.4) 26.3 23 1.01 (t, 7.4) 13.6

TABLE-US-00012 TABLE 12 NMR assignments for 18-dechloro-decarbamoyl-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00100] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.8 1 2.54 (tt, 11.5, 3.4) 39.9 2 1.73/2.20-2.13 (ddd 14.0, 32.4 11.5) 2.0/m) 3 5.22-5.18 (m) 73.2 4 3.73 (ddd 9.4, 5.4, 2.9) 70.6 5 1.85-1.77 (2H) (m) 29.5 6 1.56/2.07-2.00 (dtd 13.0, 27.7 11.2, 6.0/m) 1 168.5 2 6.02 (d, 15.1) 121.4 3 7.43 (dd, 15.0, 11.0) 146.8 4 6.53 (dd, 15.0, 11.2) 127.2 5 6.79-6.71 (m) 146.6 6 137.4 6-Me 1.96 (s) 12.5 7 6.41 (d, 10.0) 136.9 8 6.79-6.71 (m) 131.5 9 6.79-6.71 (m) 131.5 10 6.43 (d, 9.6) 128.4 11 140.7 12 2.94 (dq, 9.5, 6.6) 47.5 12-Me 1.18 (d, 7.0) 16.2 13 4.06 (dd, 9.7, 1.4) 74.1 14 4.23 (d, 1.4) 78.9 15 212.1 16 2.87/2.76 (dd 16.6, 7.6/dd, 46.6 16.6, 4.8) 17 4.58 (tt, 7.1, 5.6) 66.5 18 2.79-2.82 (2H) (m) 47.6 19 201.3 20 6.14 (dt, 15.9, 1.5) 130.8 21 7.01 (dt, 15.9, 6.3) 151.8 22 2.28 (qdd, 7.5, 6.4, 1.5) 26.6 23 1.09 (t, 7.5) 12.6

TABLE-US-00013 TABLE 13 NMR assignments for 15-hydroxy-enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00101] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.5 .sup.1 2.55 (br. t, 11.4) 38.8 .sup.2 1.74/2.20-2.14 (br. t 7.9/m) 32.4 .sup.3 5.22-5.18 (m) 73.1 .sup.4 3.74-3.70 (m) 70.5 .sup.5 1.85-1.77 (2H) (m) 26.3 .sup.6 1.61-52/2.07-2.01 (m/m) 27.6 1 168.5 2 6.02 (d, 15.2) 121.3 3 7.43 (dd, 15.2, 11.2) 146.8 4 6.52 (dd, 15.1, 11.3) 127.8 5 6.76 (d, 15.0) 146.6 6 137.2 6-Me 1.96 (s) 12.7 7 6.45-6.40 (m) 137.1 8 6.76-6.72 (m) 131.7 9 6.76-6.72 (m) 131.3 10 6.45-6.40 (m) 127.1 11 142.0 12 2.87 (dq, 9.5, 6.6) 47.4 12-Me 1.13 (d, 6.6) 16.2 13 3.97-3.93 (m) 72.2 14 3.36 (d, 8.2) 74.1 15 3.90 (ddd 10.1, 8.2, 1.8) 69.6 16 2.20/1.48 (ddd 14.3, 10.4, 1.8/ddd, 14.1, 10.2, 2.2) 40.3 17 4.24 (dt, 10.1, 2.5) 67.6 18 3.97-93 (m) 72.2 19 5.29 (t, 7.6) 75.7 19-carbamate 158.7 20 5.57 (ddt, 15.3, 7.3, 1.4) 126.1 21 5.89 (dt, 15.4, 6.5) 139.0 22 2.10 (qdd, 7.6, 6.4, 1.4) 26.3 23 1.01 (t, 7.5) 13.6

TABLE-US-00014 TABLE 14 NMR assignments for 15-hydroxy-decarbamoyl enacyloxin IIa (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00102] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.1 .sup.1 2.55 (tt, 11.2 and 3.4) 38.6 .sup.2 1.74/2.19-2.14 (ddd 14.0, 11.5, 2.1/m) 32.3 .sup.3 5.22-5.18 (m) 73.1 .sup.4 3.74-3.70 (m) 71.1 .sup.5 1.85-1.77 (2H) (m) 26.3 .sup.6 1.61-52/2.07-2.01 (m/m) 27.5 1 168.5 2 6.02 (d, 15.2) 121.2 3 7.43 (dd, 15.1, 11.2) 146.9 4 6.52 (dd, 15.1, 11.1) 127.4 5 6.76 (d, 15.0) 147.0 6 137.2 6-Me 1.96 (s) 12.7 7 6.44-6.40 (m) 137.0 8 6.75-6.72 (m) 131.7 9 6.75-6.72 (m) 131.2 10 6.44-6.40 (m) 127.1 11 142.0 12 2.87 (dq, 9.5, 6.7) 47.5 12-Me 1.13 (d, 6.7) 16.1 13 3.96 (d, 9.7) 72.2 14 3.37 (d, 8.2) 74.1 15 3.91 (ddd 10.1, 8.2, 2.2) 69.5 16 2.21/1.47 (ddd 14.2, 10.1, 2.1/ddd, 14.1, 10.1, 2.5) 40.4 17 4.45 (dt, 10.2, 2.3) 67.7 18 3.75 (dd, 7.6, 2.2) 70.5 19 4.27 (t, 7.3) 74.4 20 5.59 (ddt, 15.3, 7, 1.4) 126.5 21 5.82 (dtd, 15.4, 6.4, 0.7) 136.6 22 2.10 (qdd, 7.5, 6.5, 1.3) 26.3 23 1.02 (t, 7.4) 13.8

TABLE-US-00015 TABLE 15 NMR assignments for 5912-14 unsaturated enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00103] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 171.2 .sup.1 130.1 .sup.2 2.74-2.55 (2H) (m) 32.6 .sup.3 5.15-5.09 (m) 72.8 .sup.4 4.08-4.02 (m) 66.9 .sup.5 2.74-2.55 (2H) (m) 28.6 .sup.6 6.88 (br. s) 136.9 1 168.5 2 5.99 (d, 15.9) 121.0 3 7.42 (dd, 14.9, 11.2) 146.6 4 6.51 (dd, 14.8, 11.4) 128.3 5 6.77-6.67 (1H) (m) 146.9 6 137.3 6-Me 1.95 (s) 12.6 7 6.45-6.48 (1H) (m) 136.9 8 6.77-6.67 (1H) (m) 131.5 9 6.77-6.67 (1H) (m) 131.5 10 6.45-6.48 (1H) (m) 127.0 11 140.6 12 2.93 (dq, 9.6, 6.4) 47.4 12-Me 1.18 (d, 6.0) 16.2 13 4.08-4.02 (m) 74.0 14 4.23 (br. s) 78.8 15 211.4 16 3.04/2.83 (dd 16.9, 8.0, 2.1/dd, 17.4, 4.6) 44.4 17 4.53-4.48 (m) 66.7 18 4.05 (dd, 4.5, 1.5) 67.8 19 5.27 (t, 7.7) 75.4 20 5.55 (dd, 15.4, 7.4) 126.0 21 5.89 (dt, 15.2, 6.4) 139.1 22 2.09 (dt, 13.9, 7.1) 26.2 23 1.02 (t, 7.4) 13.5

TABLE-US-00016 TABLE 16 NMR assignments for 5m enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00104] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 180.8 .sup.1 2.25 (dt, 12.8, 5.9) 40.2 .sup.2 2.14-2.01 (2H) (m) 35.8 .sup.3 5.13-5.08 (m) 74.1 .sup.4 4.30 (q, 5.1) 73.2 .sup.5 2.14-2.01 (m/m) 33.0 1 168.5 2 6.00 (d, 15.2) 121.1 3 7.43 (dd, 15.1, 11.1) 146.6 4 6.52 (dd, 15.2, 11.4) 128.4 5 6.79-6.70 (1H) (m) 146.9 6 137.4 6-Me 1.95 (s) 12.6 7 6.45-6.39 (1H) (m) 137.0 8 6.79-6.70 (1H) (m) 131.6 9 6.79-6.70 (1H) (m) 131.6 10 6.45-6.39 (1H) (m) 127.2 11 140.7 12 2.94 (dq, 9.4, 6.9) 47.5 12-Me 1.19 (d, 6.7) 16.2 13 4.06-4.02 (1H) (m) 77.4 14 4.24 (d, 1.5) 78.9 15 211.5 16 3.05/2.83 (dd 17.0, 8.0/dd, 17.0, 4.5) 44.5 17 4.51 (ddd, 7.3, 4.4, 2.5) 66.8 18 4.07-4.03 (1H) (m) 67.9 19 5.27 (t, 7.7) 75.5 19-carbamate 158.6 20 5.55 (ddt, 15.4, 7.3, 1.4) 126.2 21 5.89 (dt, 15.2, 6.3) 139.2 22 2.14-2.05 (m) 26.3 23 1.01 (t, 7.5) 13.6

TABLE-US-00017 TABLE 17 NMR assignments for anti enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00105] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 179.4 .sup.1 2.45 (br. t, 11.3) 42.5 .sup.2 1.54-1.42 (2H) (m) 33.8 .sup.3 4.68 (ddd, 11.2, 9.2, 4.5) 74.1 .sup.4 3.58 (td, 10.1, 4.7) 72.5 .sup.5 2.28-2.22/1.54-1.42 (m/m) 33.1 .sup.6 2.02-1.98/1.54-1.42 (m/m) 28.1 1 168.6 2 5.98 (d, 15.4) 121.3 3 7.41 (dd, 15.0, 11.0) 146.6 4 6.52 (dd, 15.2, 11.3) 128.4 5 6.78-6.70 (1H) (m) 146.8 6 137.4 6-Me 1.95 (s) 12.6 7 6.45-6.39 (1H) (m) 136.9 8 6.78-6.70 (1H) (m) 131.6 9 6.78-6.70 (1H) (m) 131.5 10 6.45-6.39 (1H) (m) 127.2 11 140.7 12 2.94 (dq, 9.7, 7.0) 47.6 12-Me 1.19 (d, 6.7) 16.2 13 4.06-4.02 (1H) (m) 77.7 14 4.23 (d, 1.6) 78.9 15 211.4 16 3.04/2.83 (dd 17.2, 8.0/dd, 17.1, 4.8) 44.5 17 4.51 (ddd, 7.3, 4.4, 2.5) 66.8 18 4.06-4.02 (1H) (m) 67.9 19 5.27 (t, 7.8) 75.5 19-carbamate 158.6 20 5.55 (ddt, 15.4, 7.4, 1.5) 126.2 21 5.89 (dt, 15.3, 6.4) 139.2 22 2.13-2.06 (m) 26.3 23 1.01 (t, 7.5) 13.6

TABLE-US-00018 TABLE 18 NMR assignments for 5930/32/12-14 unsaturated enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz). [00106] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH 170.4 .sup.1 129.1 .sup.2 2.65-2.53 (2H) (m) 32.6 .sup.3 5.14-5.10 (m) 72.9 .sup.4 4.07-4.03 (m) 66.9 .sup.5 2.49-2.42 (1H) (m), 2.65-2.53 (1H) (m) 28.6 .sup.6 6.88 (br. s) 137.1 1 168.6 2 5.98 (d, 15.2) 121.1 3 7.42 (dd, 15.2, 11.1) 146.7 4 6.51 (dd, 15.6, 11.5) 127.8 5 6.77-6.71 (1H) (m) 147.0 6 137.2 6-Me 1.95 (s) 12.6 7 6.45-6.40 (1H) (m) 137.1 8 6.77-6.71 (1H) (m) 131.8 9 6.77-6.71 (1H) (m) 131.2 10 6.45-6.40 (1H) (m) 127.1 11 142.1 12 2.87 (dq, 9.5, 6.7) 47.5 12-Me 1.12 (d, 6.7) 16.1 13 3.96 (dd, 9.6, 0.7) 72.2 14 3.36 (dd, 8.1, 0.7) 74.1 15 3.90 (ddd, 10.1, 8.1, 2.1) 69.5 16 2.21/1.46 (ddd 14.1, 10.1, 2.1/ddd, 14.1, 10.0, 2.3) 40.4 17 4.44 (dt, 10.1, 2.3) 67.7 18 3.74 (dd, 7.6, 2.2) 70.1 19 4.27 (t, 7.3) 74.4 20 5.58 (ddt, 15.3, 7.1, 1.4) 130.1 21 5.81 (dtd, 15.3, 6.4, 0.6) 136.6 22 2.13-2.06 (m) 26.4 23 1.02 (t, 7.5) 13.8

TABLE-US-00019 TABLE 19 NMR assignments for 5930 + 5932_Br/H enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00107] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.48-2.59 (m) 38.8 .sup.2 1.70-1.76, 2.13-2.20 (m, m) 32.4 .sup.3 5.18-5.22 (m) 73.1 .sup.4 3.70-3.75 (m) 70.5 .sup.5 1.79-1.84 (m) 29.5 .sup.6 1.57-1.63, 2.00-2.03 (m, m) 27.6 1 168.5 2 6.03 (d, 15.2) 121.4 3 7.43 (dd, 15.1, 11.0) 146.8 4 6.54 (dd, 15.1, 11.2) 127.2 5 6.76 (d, 15.2) 146.6 6 137.5 6-Me 1.96 (s) 12.7 7 6.41 (d, 11.4) 137.0 8 6.81 (dd, 14.2, 11.5) 131.9 9 6.63-6.72 (m) 133.6 10 6.35 (d, 11.3) 136.7 11 137.5 12 2.80 (dq, 9.8, 6.7) 49.6 12-Me 1.11 (d, 6.7) 17.2 13 3.95 (d, 9.7) 72.8 14 3.36 (d, 8.1) 74.0 15 3.89-3.93 (m) 69.9 16 1.52, 1.90 (ddd, 13.9, 9.6, 42.7 2.7, ddd, 13.9, 9.8, 2.5) 17 4.12 (tt, 9.8, 2.8) 66.3 18 1.57-1.66 46.5 19 4.28 (dt, 6.5, 6.0) 70.3 20 5.50 (ddt, 15.5, 6.8, 1.3) 133.4 21 5.71 (dt, 15.4, 6.3) 134.0 22 2.06 (dq, 7.4, 1.2) 26.3 23 1.01 (t, 7.4) 14.0

TABLE-US-00020 TABLE 20 NMR assignments for BrCl enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00108] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.48-2.58 (m) 39.0 .sup.2 1.70-1.76, 2.14-2.19 (m, m) 32.5 .sup.3 5.18-5.23 (m) 73.2 .sup.4 3.70-3.76 (m) 70.6 .sup.5 1.79-1.84 (m) 29.5 .sup.6 1.50-1.62, 2.01-2.06 (m, m) 27.7 1 168.6 2 6.03 (d, 15.5) 121.4 3 7.43 (dd, 15.1, 11.2) 146.8 4 6.55 (dd, 15.1, 11.1) 127.4 5 6.76 (d, 15.0) 146.6 6 137.7 6-Me 1.96 (s) 12.7 7 6.41 (d, 11.4) 136.8 8 6.82 (dd, 13.7, 11, 7) 132.3 9 6.65-6.70 (m) 133.7 10 6.34 (d, 11.5) 136.6 11 136.0 12 2.84-2.90 (m) 47.7 12-Me 1.17 (d, 6.5) 17.3 13 4.04 (dd, 2.1, 8.1) 74.7 14 4.24 (d, 1.4) 78.9 15 211.5 16 2.84, 3.04 (dd, 4.2, 17.2, dd, 16.9, 8.6) 44.6 17 4.51 (ddd, 2.4, 4.3, 7.2) 66.8 18 4.02-4.07 (m) 67.9 19 5.27 (t, 8.0) 75.5 19-carbamate 158.6 20 5.55 (ddt, 15.0, 7.6, 1.4) 126.2 21 5.90 (dt, 15.5, 6.4) 139.3 22 2.06-2.13 (m) 26.3 23 1.02 (t, 7.5) 13.6

TABLE-US-00021 TABLE 21 NMR assignments for BrH enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00109] Position .sup.1H (ppm) .sup.13C (ppm) 1COOH .sup.1 2.47-2.58 (m) 39.1 .sup.2 1.71-1.75, 2.13-2.19 (m, m) 32.6 .sup.3 5.19-5.22 (m) 73.3 .sup.4 3.73 (ddd, 8.6, 6.2, 2.8) 70.7 .sup.5 1.79-1.84 (m) 29.5 .sup.6 1.50-1.57, 2.00-2.03 (m, m) 27.8 1 168.6 2 6.03 (d, 15.5) 121.5 3 7.43 (dd, 15.2, 11.3) 146.8 4 6.55 (dd, 15.2, 11.2) 127.4 5 6.76 (d, 15.1) 146.6 6 137.7 6-Me 1.95 (s) 12.7 7 6.41 (d, 11.8) 136.8 8 6.82 (dd, 13.6, 11.3) 132.2 9 6.63-6.70 (m) 133.7 10 6.35 (d, 11.3) 136.6 11 136.1 12 2.84-2.89 (m) 49.6 12-Me 1.17 (d, 6.9) 17.3 13 4.05 (dd, 10.0, 1.05) 74.6 14 4.23 (d, 1.4) 78.9 15 212.0 16 2.83, 2.66 (dd, 16.4, 8.4, dd, 16.6, 4.0) 47.2 17 4.20 (tt, 8.3, 3.5) 65.2 18 1.80, 1.65 (ddd, 13.8, 9.8, 43.8 3.23, ddd, 13.9, 9.3, 3.5) 19 5.23 (ddd, 9.9, 7.0, 3.3) 73.0 19-carbamate 159.7 20 5.45 (ddt, 15.5, 7.4, 1.4) 129.3 21 5.78 (dt, 15.5, 6.5) 135.8 22 2.05 (dq, 7.4, 1.2) 26.2 23 0.99 (t, 7.5) 13.7

TABLE-US-00022 TABLE 22 NMR assignments for 5927_Br/Br enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00110] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.49-2.58 38.8 .sup.2 1.69-1.77, 2.13-2.20 (m, m) 32.4 .sup.3 5.17-5.24 (m) 73.1 .sup.4 3.73 (ddd, 8.6, 6.0, 2.8) 70.7 .sup.5 1.78-1.85 (m) 29.5 .sup.6 1.51-1.62, 2.00-2.07 (m, m) 27.6 1 168.5 2 6.03 (d, 15.2) 121.5 3 7.43 (dd, 15.2, 11.2) 146.9 4 6.55 (dd, 15.2, 11.2) 127.4 5 6.76 (d, 15.2) 146.6 6 137.8 6-Me 1.96 (s) 12.7 7 6.41 (d, 11.6) 136.8 8 6.83 (dd, 13.7, 11.7) 132.4 9 6.61-6.68 (m) 133.8 10 6.35 (d, 11.3) 136.5 11 134.7 12 2.58-2.74 (m) 51.8 12-Me 1.13 (d, 6.8) 16.5 13 4.24 (td, 7.9, 3.3) 70.5 14 2.58-2.71 (m) Solvent 15 209.5 16 2.90, 2.78 (dd, 17.0, 8.0, dd, 17.0, 4.7) 50.4 17 4.31-4.35 (m) 66.3 18 4.12 (dd, 8.1, 2.4) 62.6 19 5.30 (t, 7.8) 75.7 19-carbamate 158.6 20 5.54 (ddt, 15.4, 7.5, 1.5) 126.9 21 5.89 (dt, 15.4, 6.4) 139.2 22 2.10 (dq, 7.4, 1.2) 26.3 23 1.01 (t, 7.5) 13.6

TABLE-US-00023 TABLE 23 NMR assignments for 5927_Br/Cl enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00111] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.46-2.55 (m) .sup.2 1.69-1.76, 2.14-2.19 (m, m) 32.6 .sup.3 5.18-5.23 (m) 73.3 .sup.4 3.73 (ddd, 8.6, 6.0, 2.8) 70.8 .sup.5 1.79-1.83 (m) 29.6 .sup.6 1.47-1.64, 1.99-2.06 (m, m) 26.3 1 168.5 2 6.03 (d, 15.3) 121.5 3 7.43 (dd, 15.2 11.2) 146.7 4 6.55 (dd, 15.1, 11.2) 127.4 5 6.76 (d, 15.2) 146.5 6 137.8 6-Me 1.96 (s) 12.7 7 6.41 (d, 11.6) 136.8 8 6.83 (dd, 14.0, 11.6) 132.4 9 6.60-6.65 (m) 133.7 10 6.35 (d, 11.4) 136.5 11 134.6 12 2.58-2.70 (m) 51.8 12-Me 1.13 (d, 6.8) 16.5 13 4.24 (td, 8.0, 3.3) 70.7 14 2.58-2.70 (m) Solvent 15 209.6 16 2.90, 2.78 (dd, 17.0, 7.9, dd, 17.0, 4.8) Solvent 17 4.44-4.48 (m) 66.6 18 3.99 (dd, 8.1, 2.4) 67.8 19 5.26 (t, 7.8) 75.5 19-carbamate 158.6 20 5.54 (ddt, 15.4, 7.5, 1.5) 126.2 21 5.89 (dt, 15.4, 6.4) 139.2 22 2.10 (dq, 7.4, 1.2) 26.3 23 1.01 (t, 7.5) 13.6

TABLE-US-00024 TABLE 24 NMR assignments for 5930_Br/Br enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00112] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.49-2.58 (m) .sup.2 1.69-1.77, 2.16-2.20 (m, m) 32.5 .sup.3 5.18-5.22 (m) 73.2 .sup.4 3.73 (ddd, 8.6, 6.1, 2.8) 70.6 .sup.5 1.78-1.85 (m) 29.5 .sup.6 1.52-1.61, 2.00-2.06 (m, m) 27.7 1 168.5 2 6.03 (d, 15.2) 121.4 3 7.43 (dd, 15.2, 11.1) 146.8 4 6.55 (dd, 15.1, 11.2) 127.3 5 6.76 (d, 15.1) 146.6 6 137.6 6-Me 1.95 (s) 12.7 7 6.41 (d, 11.5) 136.9 8 6.81 (dd, 14.2, 11.5) 132.1 9 6.64-6.71 (m) 133.8 10 6.35 (d, 11.4) 136.6) 11 137.3 12 2.82 (dq, 9.6, 6.7) 49.6 12-Me 1.10 (d, 6.7) 17.1 13 4.16 (d, 9.6) 73.3 14 3.45 (d, 0.7) 73.4 15 100.5 16 2.44, 1.54 (dd, 13.0, 5.0, dd, 12.9, 1.3) 43.0 17 3.49 (t, 10.1) 61.5 18 4.01-4.08 (m) 70.9 19 4.46 (dd, 10.2, 6.9) 74.9 20 5.54 (ddt, 15.4, 7.0, 1.4) 128.0 21 5.83 (dt, 15.3, 6.4) 137.8 22 2.10 (dq, 7.5, 1.3) 26.4 23 1.02 (t, 7.5) 13.8

TABLE-US-00025 TABLE 25 NMR assignments for 5930_Br/H enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00113] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.46-2.58 (m) .sup.2 1.69-1.79, 2.13-2.19 (m, m) 32.8 .sup.3 5.18-5.23 (m) 73.4 .sup.4 3.72 (ddd, 8.6, 6.0, 2.8) 70.8 .sup.5 1.78-1.83 (m) 29.6 .sup.6 1.49-1.63, 1.99-2.06 (m, m) 26.6 1 168.6 2 6.03 (d, 15.3) 121.5 3 7.43 (dd, 15.2, 11.2) 146.7 4 6.55 (dd, 15.1, 11.2) 127.4 5 6.76 (d, 15.2) 146.5 6 137.7 6-Me 1.96 (s) 12.6 7 6.41 (d, 11.6) 136.8 8 6.82 (dd, 14.0, 11.6) 132.2 9 6.62-6.65 (m) 133.7 10 6.35 (d, 11.4) 136.6 11 136.0 12 2.79-2.81 (m) 46.6 12-Me 1.17 (d, 6.7) 17.3 13 4.05 (dd, 10.0, 2.0) 74.7 14 4.23 (d, 1.7) 78.9 15 212.1 16 2.87 (dd, 16.6, 7.0) 47.6 17 2.79-2.82 (m) 47.5 18 4.55-4.61 (m) 65.5 19 201.3 20 6.14 (dt, 15.9, 1.6) 131.2 21 7.01 (dt, 15.9, 6.5) 151.8 22 2.28 (dq, 7.6, 1.5) 26.6 23 1.10 (t, 7.5) 12.7

TABLE-US-00026 TABLE 26 NMR assignments for 5930 + 5932_Br/Br enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00114] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.48-2.61 (m) 38.8 .sup.2 1.69-1.76, 2.14-2.17 (m, m) 32.4 .sup.3 5.18-5.24 (m) 73.2 .sup.4 3.73 (ddd, 8.6, 6.1, 2.8) 70.6 .sup.5 1.78-1.85 (m) 29.5 .sup.6 1.52-1.62, 2.01-2.06 (m, m) 27.6 1 168.5 2 6.03 (d, 15.3) 121.4 3 7.43 (dd, 15.2, 11.1) 146.8 4 6.55 (dd, 15.1, 11.2) 127.3 5 6.76 (d, 15.1) 146.6 6 137.5 6-Me 1.96 (s) 12.7 7 6.42 (d, 11.6) 137.0 8 6.81 (dd, 14.3, 11.4) 131.9 9 6.65-6.72 (m) 134.0 10 6.35 (d, 11.4) 136.7 11 137.3 12 2.80 (dq, 9.6, 6.6) 49.6 12-Me 1.11 (d, 6.7) 17.2 13 3.95 (d, 9.5) 72.9 14 3.36 (d, 8.1) 74.1 15 3.88-3.90 (m) 69.5 16 1.46, 2.21 (ddd, 14.0, 9.9, 2.4, ddd, 13.9, 9.9, 2.4) 41.8 17 4.29-4.35 (m) 67.6 18 3.92 (dd, 7.6, 2.3) 66.7 19 4.29-4.35 (m) 74.7 20 5.59 (ddt, 15.5, 7.1, 1.4) 130.7 21 5.81 (dt, 15.3, 6.4) 136.6 22 2.10 (dq, 7.5, 1.3) 26.3 23 1.03 (t, 7.5) 13.8

TABLE-US-00027 TABLE 27 NMR assignments for 5930 + 5932_Br/Cl enacyloxin analogue (d.sub.4-MeOH, .sup.1H 500 MHz, .sup.13C 125 MHz) [00115] Position .sup.1H (ppm) .sup.13C (ppm) 1-COOH .sup.1 2.47-2.59 (m) .sup.2 1.70-1.76, 2.17-2.19 (m, m) 32.5 .sup.3 5.18-5.22 (m) 73.2 .sup.4 3.70-3.74 (m) 70.6 .sup.5 1.79-1.85 (m) 29.5 .sup.6 1.54-1.61, 2.01-2.05 (m, m) 27.7 1 168.5 2 6.03 (d, 15.2) 121.4 3 7.43 (dd, 15.2, 11.2) 146.8 4 6.55 (dd, 14.9, 11.3) 127.3 5 6.76 (d, 15.2) 146.6 6 137.5 6-Me 1.96 (s) 12.7 7 6.42 (d, 11.4) 137.0 8 6.81 (dd, 14.3, 11.6) 131.9 9 6.65-6.73 (m) 134.0 10 6.35 (d, 11.5) 136.7 11 137.4 12 2.80 (dq, 9.4, 6.7) 49.6 12-Me 1.11 (d, 6.7) 17.2 13 3.95 (d, 9.5) 72.9 14 3.36 (d, 8.3) 74.1 15 3.88-3.93 (m) 69.6 16 1.46, 2.20-2.24 (ddd, 14.1, 9.9, 2.4, m) 40.5 17 4.27 (t, 7.5) 74.4 18 3.74 (dd, 7.6, 2.3) 71.1 19 4.43-4.47 (m) 67.8 20 5.59 (ddt, 15.5, 7.0, 1.3) 130.6 21 5.82 (dt, 15.3, 6.4) 136.6 22 2.10 (dq, 7.5, 1.3) 26.4 23 1.03 (t, 7.5) 13.8

Example 2Minimum Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) Measurements

[0461] MIC values for the compounds were determined using the broth microdilution method according to the official CLSI guidelines. Acinetobacter baumannii test strains were grown overnight in Mueller-Hinton (MH) broth at 37 C. In a 96-well microtiter plate, 50 l of serial two-fold dilutions of an enacyloxin derivative in MH broth were mixed with 50 l of bacterial suspension, diluted to a concentration of 10.sup.6 CFU/ml in MH broth. The desired inoculum density was achieved using a 0.5 McFarland turbidity standard. Following incubation for 18 h at 37 C., MICs were determined (defined as the lowest concentrations that visibly inhibited bacterial growth).

[0462] Cell suspensions without visible growth were then plated out on LB agar plates to determine the minimal bactericidal concentration (MBC). The MBC was set as the lowest concentration required to kill 99.9% of the originally inoculated 5.Math.10.sup.5 CFU/ml. All MIC and MBC determinations were performed in triplicate.

TABLE-US-00028 TABLE 28 MIC values for enacyloxin derivatives against multi-drug resistant A. baumannii strains MIC against A. baumannii Compound No. (g/ml) 1 2 2 4 3 4 4 16 5 8 6 8 7 4 8 8 9 1 10 32 11 8 12 >64 17 2 18 2-4 21 8 22 4 23 64 28 0.5 29 1 30 1 31 2 32 16 33 1 34 2 35 32