XENOCOUMACINS

Abstract: The invention relates to compounds of formula (I), some of which are derived from culture of Xenorhabdus nematophilus and Xenorhabdus luminescens. The compounds possess antibacterial, antifungal, acaricidal, anti-inflammatory and antiulcerogenic properties.

Description: - 1 - "XENOCOUMACINS" TECHNICAL FIELD The present invention relates to a new class of compounds, known as xenocoumacins, which compounds may be isolated from the culture of strains of bacteria of the genus Xenorhabdus. BACKGROUND ART Insect pathogenic nematodes of families Heterorhabitidae and Steinernematidae are known to be symbiotically associated with bacteria of the genus Xenorhabdus. It has been observed that these bacteria have the ability to inhibit the activity of other bacterial growth. International Application No. PCT/AU83/00156 (W084/01775) discloses certain lipid soluble antibiotics isolated and characterised from cultures of the genus Xenorhabdus. The disclosures of that application are incorporated herein by reference. DISCLOSURE OF INVENTION The compounds of the present invention include those isolated from the water soluble component of the culture supernatant of Xenorhabdus nematophilus and Xenorhabdus luminescens and their derivatives and have been found to have the structure of formula I. wherein R., is hydrogen, straight or branched chain. alkyl or acyl R 2 is straight or branched chain alkyl of at least 2 carbon atoms , unsubstituted or substituted by one or more εubstituents selected from hydroxy. acyl . acyloxy. halogen; R and R 4 are hydrogen, hydroxy, alkoxy or acyloxy R 5 is hydrogen hydroxy. alkoxy or acyloxy; A is -(CH,) -B. wherein m is 0. 1. 2 or 3 and B is an amino-containing radical. unsubstituted or substituted by acyloxy; or A-CH-NHR5 taken together represent a 5 to 7 me bered heterocyclic ring which may contain a further nitrogen, oxygen or sulfur heteroatom, and the pharmaceutically acceptable salts and other derivatives thereof. Particularly preferred compounds of the present invention have the structure of formulae II and III below, and are designated xenocoumacin 1 and xenocoumacin 2 respectively. III The invention also provides a process for preparing xenocoumacin 1 and xenocoumacin 2 which comprises culturing an antibiotic producing strain of Xenorhabdus nematophilus. or Xenorhabdus luminescens. in a suitable culture medium and separating the compounds of the invention and their precursors from the culture broth. Preferably, the process is a continuous process wherein culture medium is continously added to the fermenter and culture medium is continually removed from the fermenter at a rate to maintain the volume of the culture within predetermined limits. The compounds of the invention and their precursors are separated from the collected culture. The invention also provides a process for preparing compounds of formula 1 as defined above, which process comprises (a) to produce compounds of formula 1 wherein (i) R. is hydrogen, R 2 is 2-methylpropyl. R_ and R. are hydroxy. R g is hydrogen, n is 0 and A is - (CH 2^m~ B wnere ^ n i» is 3 and B is guanidino; and (ii) R χ is hydrogen. R is 2-methylpropyl, R and R . are hydroxy, R 5 is hydrogen, n is 0 I and A-CH-NHR5 is a pyrrolidinyl ring. culturing a xenocoumacin producing strain of Xenorhabdus nematophilus or Xenorhabdus luminescens in a suitable culture medium and separating the desired compounds from the culture broth; (b) to produce compounds of formula 1 wherein R.. R_. R 3 . R.. R ς . R g . n. A. m and B are other than as defined under (a) (i) oxidising a compound of formula 1 as defined above, wherein R.. R,. R,. R.. R ς , Ro-. n. A, m and B are as defined under (a) to produce an aldehyde of the formula 2 (ii) reacting the compound of formula 2 with a phosphorane of formula 3: R 4 NH*5 Ar 3 P-C-CH—A wherein Ar is aryl and R.. 5 . and A are as defined above to produce a compound of formula 4: (iii)oxidising the compound of formula 4 to produce a compound of formula 1 in which R_ is hydroxy; (c) to produce compounds of formula 1 wherein R.. R 2 , R_. R.. R 5 . R fi . n. A. m and B are as defined under (b). alkylating - compound of formula 1 as defined above wherein R. is hydrogen. R. and/or R are hydroxy. to produce compounds of formula 1 as defined above wherein one or more of R . R 1 3 and/or R„ are alkyl; or (d) to produce compounds of formula 1 wherein R.. R . R 3 , R.. R_. R g . n. A. m and B are as defined ' under (c). acylating a compound of formula 1 as defined above wherein R- is hydrogen. R and/or R. is hydroxy, or R_ is hydrogen to produce compounds of formula 1 as defined above wherein R. is acyl. one or more of R_. R and R 5 is acyloxy and/or B is substituted by acyloxy. In a further aspect the invention provides pharmaceutical formulations characterised in that the active ingredient comprises a compound of the invention. Xenocoumacins have been found to possess antibacterial, antifungal. acaricidal. anti-inflammatory and antiulcerogenic properties. The invention therefore also provides a method for the prevention or control of such conditions in a mammal requiring said prevention or control, which method comprises administering to said mammal a therapeutically effective amount of at least one compound of the invention. Preferably, the compounds of the invention are isolated from cultures of Xenorhabdus using typical extraction techniques such as liquid chromotography and subsequent extraction by an organic or aqueous solvent. Suitable culture media for the antibiotic strains of Xenorhabdus include materials containing suitable carbon and energy sources such as glucose or other carbohydrates, glycerol, or lipids, suitable nitrogen sources such as ammonia, urea, amino acids, peptides or proteins, appropriate quantities of inorganic nutrients such as phosphate, potassium, magnesium, calcium and trace elements, and preferably a source of vitamins and growth factors, e.g. yeast extract. The continuous culture processes of the invention are preferably carried out between 23°C and 37°C. most preferably at about 28°C. They are preferably carried out at a pH of between 6.3 and 7.5, most preferably about 6.8. In continuous culture, fresh culture medium is preferably added to give a dilution rate of between 0.01 hr ~ and 0.5 hr ~ , most preferably between 0.04 hr _1 and 0.1 hr "1 . MODES FOR CARRYING OUT THE INVENTION Notwithstanding other forms which may fall within the scope of the present invention, preferred forms will now be described with references to the following examples: Example 1. Culture of Xenorhabdus Xenorhabdus nematophilus strain All/1 (ATCC 53200) can exist in two morphologically different forms known as primary (1°) and secondary (2°) as reported by R.J.Akhurst in J. Gen. Micro. 121. 303-309. (1980). These forms can be distinguished by their gross cell morphology, the fact that only the primary forms elicit xenorhabdins and xenocoumacins, and by their colour when grown on 0.004% triphenyl tetrazolium chloride and 0.0025% bromothymol blue, as secondary colonies appear red since they do not absorb bromothymol blue whereas colonies of the primary form have a red core overlaid by blue. Culture 1 X. nematophilus strain All/1 (ATCC 53200) (sy biont of Steinernema feltiae All) was cultured in the following medium (5L) for 48hr. glycerol 5gL -1; yeast extract 15gL-1; MgS0 4 5mL -:L (lM); (NH 4 ) 2 S0 4 2gL~ 1 ; KH 2 P0 4 5mlL -1 (lM); 2 HP0 4 5mlL "1 (lM); Na 2 S0 4 10mlL _1 (lM). The cells were harvested by centrifugation (9000 RPM. 0.25h) and the supernatant was decanted. A sintered Pyrex funnel was uniformly packed with dry octadecyl silica (50-70um. 6cmxl2.5cm) and covered with filter paper. The solvent flow through the silica was induced by a vacuum (10 kPa) and the eluate collected in a filter flask. The silica was washed with methanol (1L). water (4L) and then the supernatant (5L) was applied followed by water (2L) and acetonitrile: ammonium acetate (1:1) (0.2M). (pH4.5.2L). This latter fraction was evaporated in vacuo to yield a crude mixture of xenocoumacins (3%. 21g). A solution of this mixture 5g) in water (20ml) was chromatographed on Sephadex G10 (84x5cm. 1650 ml) in aqueous acetic acid (0.5%) at a flow rate of 3.2ml min ~ . The eluate - 7 - was monitored continuously at 254 nm and absorbances corresponding to xenocoumacin I and II occurred at H50-l400ml and 1300-1600ml respectively. A total of 3.7g (20%) of xenocoumacins was recovered as. a brown solid. Culture 2 The following medium is found to be suitable for the culture of Xenorhabdus nematophilus ATCC 39497. Glycerol 20gL~ ; yeast extract 10gL~ ; (NH 4 ) 2 S0 4 . 20gL _1 ; KH 2 P0 4 . lOgL -1 ; MgS0 4 .7H 2 0, 2.5gL _1 ; CaCl 2 .2H 2 0.0.29gL -1 FeS0 4 .7H 2 0. 27.8mgL _1 MnSo 4 .H 2 0. S^δmgL "1 ; ZnS0 4 .7H 2 0. 14.4n.gL ~1 ; CoCl 2 .6H 2 0. O.lOmgL -1 ; and CuS0 4 .5H 2 0. 0.19mgL ~ . The medium was innoculated and growth proceeded for 6 days. The supernatant (136L) was applied to a column (150mm by lm) of Amberlite XAD-2 resin (17.1L) at 500ml.min ~ . After washing the column with 20L of water, methanol was pumped onto the column at a rate of 500ml/min and the eluent collected in 20L aliquots. 500ml of the first aqueous methanolic fraction was lyophilised extracted with ethylacetate (3 x 200ml) and then chromatographed on Sephadex G25 in water. The column (2.5cm x 76cm) was washed thoroughly with water, and then eluted with aqueous acetic acid (10% V/V) . The eluent was lyophilised. Substantially pure xenocoumacin 1 was obtained by subjecting the acetic acid fraction to HPLC on a Whatman Partisil M9 10/50 ODS column with acetonitrile: ammonium acetate (0.2M.pH4.5) 30:70 as eluant. At 4ml/min on a 254mm column the retention time was 19 minutes for xenocoumacin I. Culture 3 High yeilds of xenocoumacins 1 and 2 can be extracted from the supernatant of cultures of Xenorhabdus nematophilus strain All/1 grown on tryptone soya broth as shown in Table 1. TABLE 1 Yields of Xenocoumacins During Fermentation Time (hrs) Concentration m L- from inoculation Xenocoumacin I | Xenocoumacin II 4.5 8.7 | 0.3 23 52.6 | 40.8 28.5 72.5 | 89.4 27 72.2 | 115.9 Example 2. Recovery of Xenocoumacins The raw xenocoumacins recovered by the process of Example 1 were subjected to preparative, isocratic phase HPLC on a Whatman Partisil 10- ODS column using 0.2M. acetonitrile'.ammonium acetate (pH 4.5) in the ratio 40:60 as the mobile phase delivered at 12ml/min. The compounds of formulae II and III. xenocoumacin 1 and xenocoumacin 2 respectively were eluted at 26-30 in and 32-36 min respectively. Various compounds within the scope of the present invention were isolated as derivatives of xenocoumacin 1 and xenocoumacin 2. NMR and mass spectra data follows: - 9 - Xenocoumacin 1 and 2 13 C NMR 100.62 MHz (D 2 0.) Referenced to dioxan 67.8 ppm Carbon Xenocoumacin 1. Xenocoumacin 2 1 170.9 171.1 3 82.5 82.6 4 30.4 30.4 4a 141.2 141.2 5 120.5 120.4 6 138.3 138.3 7 116.8 116.9 8 161.4 161.6 8a 109.1 109.3 1' 22.2 22.2 2' 23.6 23.8 3' 25.5 25.6 4' 40.0 40.0 5' 50.9 50.8 7' 174.5 174.4 8' 73.7 74.4 9' 72.1 70.7 10' 55.1 62.8 11' (12') 25.7 25.8 12' (11') 26.0 24.6 13' 42.0 46.9 15' 158.00 Xenocoumacin 1 Hvdrochloride 400 MHz H spectrum ( 2 0) ref. Dioxan 3.70 wrt TMS. 8.22, 0.90, 2 x d, J6.6 Hz. l'-2'-Me; 1.41. q of d. J4a'. 4'b 13.2 Hz J4'a.3' 7.2 Hz. J4'a.5' 4.0 Hz. H4'a; 1.58. m. H3' ; 1.64. m. H12'a; 1.70. m. H4'b; 1.74. m. Hll'a; 1.79. m. H12'b; 1.83. m. Hll'b; 2.96. m. (H4) 2 ; 3.19, t. J6.4Hz, (H13) 2 ; 3.47. sextet. JIO'^'. 4 Hz. JlO'll'a 3.5 Hz. JlO'll'b 8.4Hz. H10 1 ; 4.11. dd. J9',10' 4HZ. J9'.8'. 6.1 Hz H9' . 4.20. dt. J5'.4'a 4.0 HZ. J5'.4'b 9.8 Hz. J5'.3 4.0 Hz. H5' ; 4.59. dt. J3.4a 8.1 Hz. J3.4b 4.5Hz. J3.5' 4.0Hz. H3; 6.8, 6.82. 2 x d. J8 Hz. H5. H6 7.45. t. J8 Hz, H6 Xenocoumacin 1 Hexaacetate 400 MHz X H NMR CDCl^CgDg. 3:2 referenced to TMS 0.89. 0.91, 2 x d. l'-.2'-Me: 1.20. dq. H4'a; 1.37. m. Hll'a; 1.5. m. Hllb. (H12) 2 ; 1.66. m. H3 ' ; 1.81. m. H4'b; 1.80. 1.85, 1.89.1.92. 2.10. 2.20. 6 X S. OAc; 2.53. dd. J4a.4b 14.1 Hz. J4a,3 2.8 Hz. H4a; 2.97. dd. J4a4b 14.1 Hz. J4b. 3 12.6 Hz. H4b; 3.39, q, J13.14 6Hz. J13'.12' 6 Hz. (H13')2; 4.09. dq. J3.4b 12.6 Hz. J3.4a 2.8 Hz. J3.5 1 3.5 Hz. H3; 4.24. dt. J5'.4'a 9.1 Hz. J5'.4'b. 9.1 HZ. J5'.3 3.5 Hz, H5' ; 4.43. qd. JIO',9' 7.2 Hz. JlO'.ll'a 7.2 Hz. JlO'll'b 2Hz. H10' 5.09. dd. J9',10' 7.2 Hz. J9'.8' 1.7 HZ. H9'; 5.32, d. J8',9' 1.7 Hz H8 ' ; 6.59. d. J8 HZ. 10' NH; 6.84. 6.86. 2 X d. J 8 Hz. H5. H7; 7.06. d. J 9 Hz. H6' ; 7.2. t. J 8 HZ. H7; 9.0. t. J 6 Hz. H14' . Xenocoumacin 2 Tetraacetate ~ H NMR 400 MHz (CDC1 3 ) referenced to TMS 0.93, 0.97. 2 x d. J 6.5 Hz. l'-.2'-Me; 1.48. qd. J4'a,4'b 13.5 Hz., J4'a,3' 8.5 Hz. J4'a.5' 5.5 Hz. H4'a; 1.63. dd (b). J 6 Hz. 13.5 Ha. Hll'a; 1.7. m. 3H. H3 • . H12'a. Hll'b; 1.88. qd. J4 , b.4'a 13.5 Hz. J4'b.5' 10 Hz. J4'b,3' 5.5 Hz, H4'b; 1.95. s. OAc; 2.01. . H12'a; 2.10. 2.11. 2.40. 3 x s. OAc; 2.89. dd, J 16.7 Hz. 2.5 Hz. H4a; 3.30. dd. J 16.7. 13 Hz. H4b; 3.45. q(b). _gem 10 Hz. J13'a.l2a, J13'a,12b 10 Hz. H13'a; 3.54. td. Jgem 10 Hz. J13' * b.l2a 3 Hz. J13'b,12b 10 Hz, H13'b; 4.34. m. H5'; 4.51. dt. J 12.5. 1.5 Hz. H3; 4.56. m. H10'; 5.14. dd. J9',10' 9.9 Hz. J9'.8' 1.5 Hz. H9' ; 5.21. d. J8'.9' 1.5 Hz. H8' ; 7.03. 7.12. 2 d. J 8 Hz. H5. H7; 7.52. t. J 8 Hz. H6; 8.69. d. J 8.5 HZ. H6' Xenocoumacin 2 H NMR 400 MHZ ( O) referenced to Dioxan 3.70 0.82. d. J 6.5 Hz. I'-Me; 0.89. d. J 6.5 Hz. 2'-Me; 1.39. qd. H4'a; 1.55. m. H3 ' 1.70. m. H4'b; 1.92. m. 2H. Hll'a. H12'a; 2.06. m. H12'b; 2.1. m. Hll'b; 2.96. m. (H4 2 ; 3.27. t. J 8 Hz. (H13') 2 ; 3.61. m. H10'; 4.10. dd. H9' 4.16. m. H8' , H5' ; 4.58. m. H3; 6.78. 6.81, 2 X d, J 8 HZ, H5. H7; 7.44, t. J 8 Hz. H6. Xenocoumacin 2 Monoacetate X H NMR 400 MHz (CDC1 3 /D 2 0) referenced to TMS 0.93, 0.96. l'-.2'-Me; 1.50, qd. J 13.5. 8.6. 5 Hz. H4'a; 1.68. m. H3'; 1.79. m. H4'b 1.88. m. H12'a; 1.91. m. Hll'a; 2.04. m. H12'b 2.12, S. OAc; 2.27. m. Hll'b; 2.83. dd. J 16.7, 2.5 Hz. H4a; 3.11. dd, J 16.7, 13 Hz. H4b; 3.53, m, H13'a. H13'b; 3.70. dd. J 7.8. 4.4 HZ. H9' ; 3.90. d. J 7.8 Hz. H8 ' ; 4.21. m, H10' ; 4.35, m, H5'; 4.60. dt. J 12.7. 2.4 Hz. H3; 6.71, 6.90, 2 x d. H5. H7; 7.42. t. H6; 7.9. d. 9Hz. H6 ' 10.8. x. 8-0H. ■ - 14 - MASS SPECTRA Xenoucoumacin 1 Hexaacetate Negative Ion Chemical lonisation (isobutane, gold probe, 200°) m/z: 717M " (25), 675 (52) (M-CH 2 CO) . 657 (42). 615 (52). 403 (5), 361 (20). 254 (10). 163 (38). 150 (100). Electron lonisation. 70 EV, 200°. gold probe m/z: 717 M + (2). 702 (4). 675 (15). 658 (10). 632 (10). 616 (10). 512 (20) 470 (15). 341 (5). 267 (28). 184 (90). 170 (28). 112 (30), 86 (50). 70 (100). Observed Mass 184.1076. Calculated for CI8H-14.N.3O2- 184.1086 Positive Ion Chemical lonisation (ammonia, gold probe, 200°) m/z: 718 (M + H. 75). 676 (MH-CH 2 CO. 100), 658 (15) 634 (10). 617 (12). 593 (10). 557 (7). 513 (15). 471 (12). 267 (22). 184 (40). 170 (15). 112 (15). Xenocoumacin 2 Hexaacetate Negative Ion Chemical lonisation (isobutane. gold probe. 200-°) m/z: 573 (M- H) . 531 (M - CH 3 CO. 100). 514 (8). 454 (18). 412 (12). 394 (12). 265 (10). 222 (10), 163 (8). 130 (10). Electron lonisation (70 EV, gold probe. 200°) m/z: 574 (M + . 10). 369 (10). 242 (11). 196 (10). 154 (12). 135 (5). 112 (35). 70 (100). Positive Ion Chemical lonisation (ammonia, gold probe, 200°) m/z: 575 (M + H. 100) 533 (MH-CH 2 0, 12). 370 (10). 242 (8). 196 (12). 154 (8). 112 (15). 70 (17). Xenocoumacin 1 Fast Atom Bombardment: Observed Mass 464.2574 Theoretical Mass for C _H Λ N_0. (M - H) 464.2509. Example 3. Production of Derivatives The following synthetic pathways serve to illustrate how derivatives of xenocoumacins may be produced. Ar is an aryl and A.R 4„ and R c 5 are as defined above. As will be understood by those skilled in the art. the stereospecificity may be altered or controlled by altering the oxidant used. e.g. H O /Se0 2 will confer a different stereochemistry on the vicinal diol to Os0 4< R-. R, and R 4 of formula I may be varied by known processes of alkylation and acylation as per the following reactions: 1) Alkylation wherein R 2) Acylation wherein R is Example 4. Antiulceroqenic activity The antiulcerogenic activity of the xenocoumacins of the present invention was demonstrated by the following tests: Test 1 Male Wistar rats were dosed perorally (p.o.) dosed with varying amounts of xenocoumacin 1. The animals were then exposed to ulcer inducing stress conuitions. killed and examined for ulcer induction. The results were compared to a control using the Wilcoxon rank sum test. Significance is taken as p< 0.05. Xenocoumacin 1 was shown to be 74% effective when a dosage of 25 mg/kg.po was administered, but only 8% effective for a dosage of lOmg/kg.po. Test 2 Test 1 was repeated using xenocoumacin 2 , which was found to be 70% effective for a dosage of 25 mg/kg.po.. 61% effective for a dosage of lOmg/kg.po and 26% effective for a dosage of 5mg/kg.po. The xenocoumacins of the present invention may be used in the treatment of bacterial or fungal infections or in the treatment or prophylaxis of inflammatory diseases or ulcers. The compounds or mixtures of them may be administered in standard pharmaceutical formulations, in admixture with known pharmaceutically acceptable excipients. adjuvants and diluents. Formulations of the invention may also contain other active substances. Example A Xenocoumacins 1 and 2 can be used independently or in admixture as the active ingredient in the manufacture of the following pharmaceutical formulations: (a) Tablets 1 tablet contains Active ingredient 200 mg Microcrystalline cellulose 155 g Maize Starch 25 mg Talc 25 mg Hydroxypropylmethylcellulose 20 mg (b) Capsules 1 capsule contains Active substance 100 mg Maize starch 20 mg Lactose 95 mg Talc 4.5 mg Magnesium Stearate 0.5 mg The xenocoumacins of the invention may also be administered topically or intravenously. Preferred dosage rates are within the range of O.lmg/kg to 50mg/kg when administered periorally. and 0.01 to 20 mg/kg when administered intravenously. Topical formulations can contain up to approximately 10% active substance.

Claims: CLAIMS Compounds of the formula I wherein R- is hydrogen, straight or branched chain. alkyl or acyl R 2 is straight or branched chain alkyl of at least 2 carbon atoms, unsubstituted or substituted by one or more substituents selected from hydroxy. acyl. acyloxy, halogen; R,3 and R4, are hydrogen, hydroxy. alkoxy or acyloxy R 5 is hydrogen hydroxy. alkoxy or acyloxy; and A is -(CH-2)m-B. wherein m is 0. 1. 2 or 3 and B is an amino-containing radical. unsubstituted or substituted by acyloxy; or A-CH-NHR5 taken together represent a 5 to 7 membered heterocyclic ring which may contain a further nitrogen, oxygen or sulfur heteroatom. and the pharmaceutically acceptable salts and other derivatives thereof. 2. A compound of formula 1 as defined in claim 1, wherein R. is hydrogen. R_ is 2-me hylpropyl. R and R . I are hydroxy, R5 is hydrogen, n is 0 and A-CH-NHR5 is a pyrrolidi ' nyl ring. 3. A compound of formula 1 as defined in claim 1. wherein R. is hydrogen, R 2 is 2- methylpropyl. R and R. are hydroxy. R c -> is hydrogen, n is O. A is -(CH-.)m-B wherein m. is 3 and. B is. guanidino. 4. A compound of formula 1 as defined in claim 1. wherein R. is acetyl, R 2 is 2-methy propyl. R 3 , 4 and R ς are acetyloxy. n is 0. A is -(CH 2 ) m -B wherein m is 3 and B is l.3-diacetyloxyguanidino. 5. A compound of formula 1 as defined in claim 1. wherein R is acetyl. 2 is 2-methylpropyl. R 3 , R 4 and I R5 are acetyloxy, n is 0 and A-CH-NHR5 is a N-acetyloxypyrrolodinyl ring. 6. A compound of formula 1 as defined in claim 1. wherein is hydrogen, R is 2-methylpropyl. and R 4 I are hydroxy, R5 is acetyloxy. n is O and A-CII-NHR5 is an N-acetyloxypyrrolidinyl ring. 7. A process for preparing compounds of formula 1 as defined in claim 1. which process comprises (a) to produce compounds of formula 1 wherein (i) R- is hydrogen. 2 is 2-methylproρyl. 3 and 4 are hydroxy. 5 is hydrogen, n is O and A is - (CH 2^m ~B wherein m is 3 and B is guanidino; and (ii) χ is hydrogen. R 2 is 2-methylpropyl. R 3 and R 4 are hydroxy. R-. is hydrogen, n is O I and A-CH-NHR5 is a pyrrolidinyl ring, culturing a xenocoumacin producing strain of Xenorhabdus nematophilus or Xenorhabdus luminescens in a suitable culture medium and separating the desired compounds from the culture broth: (b) to produce compounds of formula 1 wherein R_, R 2 « R 3 , R 4 . R c . R g . n. A. ii: and B are other than as defined under (a) - 21 - (i) oxidising a compound of formula 1 as defined in claim 1. wherein R. ^ . R . R 3 . R 4 . R Rfa e , n. A, m and B are as defined under (a) to produce an aldehyde of the formula 2 (ii) reacting the compound of formula 2 with a phosphorane of formula 3: R 4 NHR 5 Ar 3 P= C-CH—A wherein Ar is aryl and R 4 « R 5> and A are as defined in claim 1 to produce a compound of formula 4: (iii)oxidising the compound of formula 4 to produce a compound of formula 1 in which R is hydroxy: - 22 - (c) to produce compounds of formula 1 wherein R . R . 3 . R 4 . R_. R g . n. A. m and B are as defined ' under (b), alkylating a compound of formula 1 as defined in claim 1 wherein R, is hydrogen. R, and/or R. are hydroxy, to produce compounds of formula 1 as defined in claim 1 wherein one or more of, R.. R 3 and/or R 4 are alkyl; or (d) to produce compounds of formula 1 wherein R_, R 2 « R 3 , R 4 . Rg. Rg. n. A, m and B are as defined under (c), acylating a compound of formula 1 as defined in claim 1 wherein R., is hydrogen. R, and/or 4 is hydroxy. or 5 is hydrogen to produce compounds of formula 1 as defined in claim 1 wherein R 1 is acyl, one or more of R , R 4 and R 5 is acyloxy and/or B is substituted by acyloxy. 8. A process as defined in claim 7. wherein the xenocoumacin producing strain is Xenorhabdus nematophilus strain XQ1 (ATCC 39497). 9. A process as defined in claim 7. wherein the xenoconmacin producing strain is Xenorhabdus nematophilus strain All/1. (ATCC 53200) 10. A process as defined in claim 9. wherein the medium is tryptone soya broth. 11. A compound of formula 1 as defined in claim 1. whenever produced by the process claimed in claim 7. 12. A pharmaceutical composition, which comprises at least one compound of formula 1 as defined in claim 1 together with a pharmaceutically acceptable carrier or diluent therefor. 13. A method for the prevention or control of infectious disease in a mammal requiring said prevention or control, which method comprises administering to said mammal an effective amount of at least one compound of formula 1 as defined in claim 1. 14. A method of prophylaxis or treatment of ulcers in a mammal requiring such prophylaxis or treatment, which process comprises administering to said mammal an effective amount of at least one compound of formula 1 as defined in claim 1.

Inventors: GREGSON RICHARD PETER, MCINERNEY BERNARD VINCENT, GREGSON, RICHARD, PETER, MCINERNEY, BERNARD, VINCENT
Applicants: BIOTECH AUSTRALIA PTY LTD, BIOTECHNOLOGY AUSTRALIA PTY. LTD.
Application year: 1985
Main classification: C07D311
IPC codes: C07D311, C07D405, C12P17