MXPA98002749A - Orthosomicinas novedosas isolated from micromonospora cardona - Google Patents

Abstract

Orthomyomycins have been isolated from the fermentation broth of microorganisms Micromonospora Carbonacea see Africana designated SCC 2146. These compounds are antibacterial agents. Compositions and methods for the treatment of bacterial infections are also described

Description

ORTOBOMICINAB NOVEDOSAS AISIADAS A PARÍ IR DF. MICROMONOSPORA CARBONACEA COMPENDIUM OF THE INVENTION Comets I, A,, D, E, F, O, H and J were isolated from] > Fermentation of the fermentation of the Mir.romonoipciri Carbonac-ía at * Afí irana de njip to SCC 2146. These? -nmμnμ. ,?) s fuer n? dí n t i. f i na o? like ort-nails. These oment --- »n g ntes ni ibac: mpano = > . A coinfusion was obtained from the dollar, the letter A, the agent, * * * tnl was not more powerful *. the invention refers to novel, novel, A, B, C, D, E, F, G, H, and J, and their components and A compositions containing such compounds. This invention also relates to a fermentation broth of 3-chromium, and the component parts of the MO that can be obtained by cultivating a pure culture of Mu rom, not by,. Oarbonacea wav A f i an. The invention relates to microorganism Mi ro onospora Ca bona to African var. Another asp ct d the present invention is foc-? It is also the result of the cultivation of a product produced through the cultivation of a crop of My cro-3pora C.irbun. African fruit in a water nutrient with a correct pH and temperature that has assimilable sources - > carbon and nitrogen b:?.? or cond i > Aerobic substances submerged in the production of a composition of matter having a substantial antibiotic activity.A major component of the present invention is antibiotic 11-384, component 1 , in accordance with the res. in USP 4,597,968, which is incorporated herein by reference. (Another major component of the culture is the corresponding nitroso analog.) However, the present invention recites other competes. As described above, the invention is also related to an antibiotic composition comprising a pharmaceutically acetablized vehicle and an ibidically effective one or more selective co-products within the scope of the invention. A group consisting of compounds A, B, C-D, F, 8, H, and J. The present invention also relates to a method for the I, rt, and ami-t of a bacterial infection comprising the administration of a quantity an ib ló It can be used effectively for one or several sci cient components within the range of the comps A, B, C, D, E, F, G, H and!. DESCRIPTION OF ITS TIGURES l -3 s f i gu vas 1, 7, 4,, 6, 7 and 8 are NMR spectra of p ro t n pa va l = > omitted A, B, E, F, S, H, and I, respectively »Figure 7, is a NMP proton spectrum for a cc mec. mpo í > C and c. DETAILED DESCRIPTION OF THE INVENTION FERMENTATION OF THE MICROORGANISM A final congel 3 of 1 «? | _ Of icromonospora carbon ea - > African Species, strain PF-3 was transferred into a 300 ml Erlenmeyer shake flask containing 100 μL of seeded medium. The average composition (g / L) was the following network extrusion (Difco) 3, Tryptone (D? Fca) 5, Cerelose (CPC, 2001) 1, dei. pineapple potato (Avebe, WPD-650) 24, yeast extract (Universal, Tastone) 5, potassium carbonate (Pfi r.er, Albaglos) !, and si icone foam remover (Union Carbide, SAG-471, 30% suspension) 0.3 mi / L. The bottle was incubated for 48 hours at a temperature of 30 ° C with shaking (3 rd i'pm, displacement and 2.54 &ut) and 30 ml. of the culture were transferred to bottles of 6;, 1, C-ida one containing 500 ml of the same one was thrown away. After 48 hours of incubation as before, the culture of 31 was transferred in n termenter by inoculation containing 300 L of the medium for an additional 24 hours of culture. Finally, the content of the inoculum fermentor was transferred to a large fermenter containing 10,000 L of production medium. The composition of the production medium (g / l.) Was as follows; e: - trac Yeast 5, peptor > 3 e meat1 (Marcor, type PS) 6, Cr i se 22, infusion of m i z in pol o (Marcor) 2, from: trina le papa 0, boiled linseed oil (- le ^ n tr i) 4 , carbonate) de cali 10 4, cobalt chloride to «6M20 (Mal 1 cl * rod) 0.002, silicone foam remover 0.5 mL / L. The fermentation was carried out at a temperature of 36 ° C for 1 <At a time of 11 ° under the introduction of 11 ° C and stirring, keeping the oxygen free between 5 ° and 1 ° of saturation, the fermentation will be carried out with 2.27 - 6.82 cubic meters. It is per minute of flow during pouring: 12 to 30 hours per day, and the fermentation medium was cooled to about 25 ° C. Half of the fermentation broth was transferred to a vessel. = .ep =? Fetched, shaken and adjusted with 2N NaOH to ll gar at 1 p> H of 10.5, 200 l of ream XAD-7 (Rohm Ha-is absorbent polymerase of non-functional acrylic ester) was charged. The fermentation broth was stirred at 0.5 hour. The pH was lowered to 9.25 and shaken for 3.5 hours, the pH was lowered adonly to 7.00 and the beef was separated from the broth by sieving. Water was used from the water to remove the resin to remove the carbon and mycelia, and the middle half of the water cycle was processed in the same way. 300 l of ream adsorbed to a 500-liter tapered column containing 1 OO 1. of water, of deion. The resin was washed with the flow up e. After bathing the aqueous level to the resin bed, the reed was washed with downflow with 900 L of deionized water. The antibiotic was eluted from the descending flow of res to medi-before the loading of the column nn 9 0 l of acet or of ethyl, pre-washed with 140 L of sodium bicarbonate buffer < "*, 1 M, adjusted =? 1 pH of 8 (on hydrolyzate of .odium) at the rate of t L / mlu. The eluted product was collected > - n cuts of 150 to 200 l. Short films containing antibiotic complex were co nadon and ekt frayed rum 1. sodium phosphate onoLiSic IM, just at 1 pH of 0 with sodium hydride and then with 0 or 0 l de ayua desiofii < sda. The ethyl acetate layer was concentrated in vacuo at a temperature below lo-> 3? r to one tenth of the original volume (Approximately 5 < > ) with azeotropic distillation of residual water. The concentrate was precipitated in 100 l of heptane (2 volumes). The precipitate was filtered and dried at a temperature of approx. 25 ° C in a vacuum oven using a nitrogen curve to provide 5.2 to 5.5 Lg of raw material (2.6 to 3.1 Lg of antibiotic complex). OXIDATION With stirring, 10 to 11 μg of the crude antibiotic complex was dissolved in 80/20 ac tates of ethyl-acetorw to remove the nitrous component of the antibiotic 13-384 complex. Two times we obtained sodium bicarbonate and 50 g of cata- lling and ethanol were added. 5.5 L of hydroperoxide of tert-butyl (3 M solution in 2.2, 4-trone, 1 lpntane) was added slowly over 0.5 hour while the temperature was maintained within 20% and between 25 and 50 minutes. 30 < , 0"The progress of the oxidation was monitored by HPl C (cro tog of the high pre-ion liquid). Additional catcher was added when laughing and the addition of the pi reaction continued until the end of the reaction. INITIAL PURIFICATION Approximately 2.5 kg of crude oxidized material was dissolved in 9 L of ethyl acetate and agitated in the column of urii column d 3.048 m by 0.3 48 m in diameter packed with 70 P of bare irregular silica gel (from 70 to 250 μ) in tsopropyl acetate (the sol may also be 8/20 acetate and 11 / hep auo). the column was eluted with isopylacetate or il at a rate of 5 to 7 l / minute at 24 ° > 09.5 Lg / m2 on atmospheric pressure. Fractions 1, 2 and 3 were collected as 200 L cuts. The antibiotic components were monitored by HPL and TLC. The solvent used for the operation of TLC was 9% methyl alcohol chloride. The fractions from 5 to 11 that were enriched in the main antibiotic component were combined and concentrated in an approximate way, L less than 30 ° C. The main cut was isolated by the addition of concentrate. to two volumes of heptan? with agitation. The precipitate was filtered and dried in a vacuum oven at a temperature of about 25 ° C with a nitrogen purge to obtain 1.2 l-g of product. The F's? actions 3 and 4 of be cut. which were enriched in impurities were processed., similarly p-tra obtain 0.4 g of product. The scission action of cabbage 3 12 to 15 that were enriched in? mpnue, were processed in a similar way to obtain 0.2 tg of product. Additional Purification of the Co-light Antibiotic The purification was carried out in a procedure i roma togr >; f i co in two steps. First the head cuts (ID 33285-104-2) and tail (ID 33285--104 -1) obtained from the initial purification were chroma tograt i pts and medium pressure conditions with the diol bound silica (medium go eg 40 res. -63 μ). the column was equilibrated with a ternary channel of CH2C 12 shepi year: MeOH (60: 4 (-> v2 v / v / v> at a flow rate of 3 <"-50 ml / in. with gs nitrogen After the application of the sample, the mobile phase was collected and discarded on 2.4 L-2.8 I, the mobile phase was then adjusted and a friction was collected based on the volume of ca to the column of d. containing new compounds were then puri fi ed by chromatography of liquid from the above liquid: 51 pf semipreparated in PVA-silicon silica func. ), with a peak collection based on the UV signal monitoring at 265 nm.For individual component pu tifications, minor adjustments of the mobile phase were made (see diagram l) E j em 1 o No 1 dissolved five grams of enriched tail cut (ID 3325R-104 ~ 1) p 20 t of 0H2012; MeOH (: v / v) and applied to 200 g (approximately 400 ml.) of sil? a pt re-bound ludicidal uni an diol (40-63 μ, irregular mei s) contained in a glass column (6O0; 50 milli, 1 «IR L). The diol column conditioning step before the 13 application of the sample involved passage of MeOH II followed by 1.2 L of initiating mobile phase CH2C I heptan MeOH (60: 40: 2 v / v / v /). After the application of the sample, 2.4 I of mobile phase was collected after that time the proportion of phase1 mobile was adjusted to 75: 25: 2 and maintained for an additional 8 1. The individual fractions were obtained and evaluated in order to determine the presence of minor components by means of analytical HPl under the following conditions? Se »_r < i > t? .aa s PVA-Sil, 5 μ, 15 cm; - 4.6 m CH2Cll: eOH (98: 2) / 20 minutes. Complex 1 provided 20 utg of material containing 15% of a new Compound, which is less than anti-ion, 1 -38, component 1. Complex 1 was piurifed? ad 11.1 onal by dissolving e 3O-40 mg of the sample in 0.5 ml of CH2C12: MeOH (96: 4 v / v) and by injecting into a column PVA-S? l - = em? pret ra t i va (250 x 2C <mm) equ 111b with CH2C 12: MeOH (97.5: 2.5 v / v). A flow rate of 12 ml./mm yielded 5 mg of the desired material within a window and elution time of 1-14.2 minutes. After four additional injections were made, a total of 24 mg of Compound A < , 98 * / »pure). Complex 2 (100 m) was purified additionally by using the same HPIC conditions, which would be pre-prepared and modified above unless it was empiled. a CH2C12 solvent system: he tano: MeOH (78: 20: 2, v / v / v) as a physical phase. Two pure components were obtained, the Compues or E1, 5 m and the Compound F (9.5). However, the first component (2.4 m) was identified as an ez l of two co-components, Compound C and Compound D, in ba ^ a an-? »! i es e d os spec osc os p 1 os. Ex ml No. 2 Five grams of enriched head cut (ID 33285-104-2) were dissolved in 25 mL of CH2C12: MeOH (96: 4 v / v) and applied to a column of linked silica gel c? n a recollection of 200 g. Recycling involved this fine of 1.5 i. of MeOH, aeyui, or by 1.5 I. of phase mó il 1 m to CH2C1 shep, 3 no: Me0H (60: 40: 2 v / v / v). Desp > of the aptl ic.ae of the sample, re > I want 2.8 l of mobile phase 10 and the effluent was discarded, the mobile phase was adjusted to 75: 25: 2 (v / v / v) and fractions of 40O ml were collected .. Based on ana lytic HPLC, e four fractions were combined as complex 3, and were obtained 3 ¿> n mg of a yellowish powder after evaporation and ion, (The fractions were combined as well as compile 4). The HPC1 analysis of the compile jo 3 is PVA-Si 1 indicated two peaks at times of measurement prior to Compound A. Studies of the aptitude for ra >; The preparation atography led to the selection of a binary solvent system composed of n-but i 1 l aride MeOH that revealed the presence of a third entity. Then, approximately 40-45 m of complex 7 were dissolved in 1.0 ml of CH2C 1: MeOH (96: 4 v / v) and dissolved in a PVA ~ S? L column of se ipreμ ri in (250: <) 20 m) equilibrated with n-bu t 11 loride: MeOH (93: 7 v / v). The flow rate was 15 mi / min and the detection of UV rays was 265 nm. If components are components, the first two components (9.5 mg and 11.5 mg) were unstable and could not be identified-1. Only the third component (39 m) was identified as Component B. purified ad? I n 3 Complex 4 (61 m) by means of method > ie HPLC of sem i f > rep > ara > . Modified ion similar to the con ions described above with a mobile phase different from CH2 l2s eOH (97.5: 2.5, v / v). Compound B (6.1 mg), Compound H (8.7 g) and Compound T (27.5 mg) were obtained from three pure components. SCHEME 1 Cot * te de col (ID 3328 - 104 - 1) Silica urti da with d i o1 < CH2C12: Hep t. : MeOH) (6.0: 40: 2) Sr step lens / J 5.) Ccuíip 1 e o 1 Com l o 2 PVA-il P A-Si 1 CCH2C12sMeOH) (CH2C12: Hep Me0H) (97.5: 2.5) (78: 20: 2) Compound A Compound Compound E Compues F (Compound C + Compound D) DIAGRAM 1. orti, i r? Ac i órt Frontal Cut (ID 33285-- 1 -2) Silica u i a with diol (CH2C12 sHe .: MeOH) C C this 7 PHYSICAL-CHEMICAL PROPERTIES All the competitors were obtained as white polymers after the removal of solvents. The compounds are soluble in methanol, or; < gone from d i me 111, ethyl acetate, acetone and chloroform; partially soluble in diethyl ether, loromethane and 1- lorobutane; Insoluble in oil, ether, petroleum and water. The physical-chemical properties and the specific nature of these > . or pipettes of the invention - = e r su n e Table 1.

TABLE 1. P rop f e s i nc y and m o n e = spec ples of Comp iueations of T he Com m on Compue a Compue to B Compound C (l fa) 20D - 48., "-45.2" 3 í c 0.1, M.iOH) UV 211 211 211 mx »(nm) 268 265 268 305 307 IP ma: 3454.3939, 1734 3430.2940, 1729 3447,2939,1730 (cm-1) 1653,1544,1457 1622,1544,1456 1652, 1 23,1544 1384.12 fvC, 1128 1684.1258, 1103 1455.1 84.1343 1 65, 1030, 90 i or: 1528,1102.1 37 Weight 1663 162"1645 m or 1 et u 1 ar E piee tt or 1664 (M + H) + 1628 (M? H) + 1646 (M + H) i- e mass FAB F or rmu l C7OH96N038C1" C70H95N038C12 C71 H1O1 N038C 1 ol c ui t 13C NMP 120.9 ppm 120.9 ppm 120.9 ppm (CDC13) b 118.9 ppm 118.9 ppm 119, ppm 1H NMP FI6 1 F1 7 FIO 3c (CDC13) Compound D Compound E F αalfa Compound) 20D at -47.2 ° -47.0 ° (c 0.1, MeOH) UV 211 211 211 ma. nm) 268 268 264 307 305 305 IR max. 3447, 293, 1730 3431, 2939, 1729 3477, 2936, 1728 (cm-1) 1652,1623,1544 1652.1 23.1544 1652.1 22.1544 1455.1384,1343 1455.1384,1257 1457.1384,1341 1258, llo2, l037 11 5, 1 37 2060, 1128.1 36 Weight 1643 1581 1595 Mo1 r u1 ar Espe ro 1644 (M- »H) 1 82 (M + H) + 1596 (MU ~ l) + of mass F? B Formula C71 H99NQ38C1 C69H96N038CI C70C98N038C1 mol cu 3 ar 13C NMP 120.2 ppm 120.2 ppm 1 0.1 ppm (CDC13) b 118.9 ppm 119.0 ppm 118.9 ppm 1H NMR FI8 3c FIG 4 F1G 5 (CDC1 Z) Compound 8 Compound H Compound 7 (alf3) 2 D -45.5 ° 45.8"-72.3 ° Ic 0.1, MeOH) UV 211 210 m;. <265 265 30, or IR max 3434.2939.1734 3430.2937.1734 459.29; , 16 (cm-1) 1652,1622,1544 1653, 1 23, 1543 1 2, 1452, 1315 1454.1 84,1257 1 57,1385,1345 362,1173,1101 11? , 1038 1036 1068, 1034.99: Weight 1615 1645 m or 1 ec u 1 a r Espe o í M-i) • * - 1646 Í + H) + (M + H) + of mass F? B Fò'-mul a e? N038C1! C70H9"> C12 C27H36017 ol C r 13C NMP 120.9 ppm 1 1.0 ppm (CDC1) b 119.0 ppm 119.1 ppm 11.0.0 ppm Í NMP F1G 6 F? 6 7 FIO 8 (CDC13) 3. The optical rotation was not measured due to the number of components. b. Only two major chemical shifts of distinct ortho-ester carbons were presented in their list. c. Figure 3 shows the resen i of a sample of two compounds. d. Compound J possessed only an ortho ester functionality. ECTRUCTUPAI DETERMINATION OF COMPOUNDS OF INVENTION The structures of the compounds were elucidated on the basis of analysis of specific data, including ultraviolet (UV), infrared (IR) analysis. , Espe tromet of Fast Atomic Bombardment Mass (FAB-MS), proton nuclear magnetic resonance and bonus methods - 13 HH and 130 NMR) »This» components were carat lep ados such as ant ib? / > t i-os related to evermrto ic i na novelty. Data from anAli is with magnetic resonance r r 13C two ort > - esters and ports appear in a list in Table 1. The spectral data of 1 H nuclear magnetic resonance of indi iduals are illustrated in Figures 1-8, respectively. It assigns ions, of some protons and important carbons the IAPT-bound proton test was achieved immediately - the oscillating spectacle of the two-dimensional nuclear spectrum., the correlation of multiple-bond ion ler onu lear (HMBC) and multiple-quantum coherence experiments (HMQC), as well as by means of a direct comparison of spectral data with the antibiotic (13 ~ 384-c omponent -1, evernmomie ina) claimed in the USP document 4,597,968 co or a reference standard. EXAMPLE 1 Elution of the structure Compound A was carried out by mass analysis and specific nuclear magnetic resonance data. Da espiec. The mass of FAB showed an increase of 34 molecular weight master compared to the reference sample (13-3R4- component 1, evern i nom i 'm-t). A group of molecular ions containing trie parrot was observed in the FAB mass spec. Both observations revealed the presence of the atom of. In the compound A, the binding of this additional chlorine atom to the fragment of e-ater to rom at the right-hand side of the molecule was determined on the basis of an ana.li.-iise secondary fragmentation in comparison with the reference sample 13 ~ 384 ~ - component-1, evernomi c ma). (DIAGRAM 1). However, the mass spectral data could not locate the exact position of the chlorine atom in the aromatic ring. The position of this chlorine atom C-58 was determined originally on the basis of the spectral data of nuclear magnetic resonance, which indicates a strong correlation of pro - - lt; £ > 0 and methyl pratortas ~ 62 in NOESY experiments, and one 11 v idad between pr? Toror? -60 and ol 11 lcarbon-62 in HMBC. Therefore, the structure of Compound A * e was determined as illustrated in DIAGRAM 2. Through the use of the same methodology, the structure of other combinations was also elucidated and illustrated in DIAGRAM 2, Compound C and Compound H are illustrated in DIAGRAM 3. It will be noted that Compound 7 was stretched as a relatively small disaccharide linked to one. portion of aromatic and cyclic ester through ortho-ester functionality as illustrated in FIG. 4 below. DIAGRAM 1 DIAGRAM 2 everninomi .i na- * H OH 0H, H OMe Cl Me Compound A Cl OH OH, H OMe Cl. Me Compound BH OH = 0 OMe Cl Me Compound DH OMe 0H, H, OMe Cl Me Compue to EH OH 0H, H OH H Compound FH OH 0H, H OMe H Compue to GH OH 0H, H OMe Cl H * Evernomycin is the antibiotic 13-384, component 1 as presented in USP 4,597,968 Italic capital letters identify rings in the compounds of the invention. The structures for compounds C and H are presented below. Diagram 3 U Compound C P.7 = CH3; R8-CH3 Compound H R7 = C (0) H; R8 = H.

The structure will piace the. Compound J is illustrated below in Diagram 4. DIAGRAM 4 BIOLOGICAL PROPERTIES OF THE COMPOUNDS OF THE INVENTION The minor components were tested to determine their activity based on an agar disc diffusion protocol. For compounds, it was dissolved at a rate of 1 mg / mL in CH2C12 / MeOH (95: 5 v / v) and a 10-fold dilution was made in the same vehicle. 20 microliters of each concentration was transferred to a standard 8 mm disc and allowed to air dry for 30 minutes. Each set of disks was placed on agar seeded with Staphylococcus aureus in two pH (7/8) and incubated overnight at a temperature of 35 βC. The zones of the inhibition channels are then given as the diameter of the inhibition circle and d = m in millimeters. The results are shown in the following table: C aity 20 μg 2 μg 20 μg 2 μg Everninomi ina 21 19 26 23 Component A 20 2? 26 23 Component B 14 12 17 13 Component C / D 2 16 22 17 Component E 18 16 22 17 Component F 17 16? 0 17 Component G NT NT NT NT Component H NT NT NT NT Component TO 10 0 As used here , NT means not tested the almost equivalent potency of Compound A n everní no ii na was documented adi ci nally in a di 1 u > What is the U? t ro vec is "The live antibiotic activity of the compounds of the invention can be constant in mice through the administration of subcutaneous ion. This invention can be carried out by employing pharmaceutically acceptable compositions comprising a pharmaceutically acceptable carrier and one or several others selected from the group consisting of A, B, C, D, E, F, G , H and 7.

As such, antibiotics can be administered with any v * -hi > It can be administered in an appropriate manner and administered orally, preferably, or topically in several formulations. Pava oral administration, the antibiotics of invention may have the form of tablets, capsule, elixi is and if my the re-a. Tablets and > capsules may contain - '? p > ? entity = 5 as for example starch or the bear; the liquid forms can have an ag »» > Or prayers or flavor?. * ard e. The topical ions can be in the form of creams, hydrating ointments or hypodermic ointments, or aqueous or non-aqueous emulsion lotions. Typical vehicles for these formulations are water, oils, fats, polyesters and polyols. Formulations p rentera! s, for example, injectable dosage forms are usually liquid such as solutions or suspensions and the typical vehicles are distilled water or solid solution. The dose to be administered) in > Any particular form of dosing. What will depend on several factors such as erythropathy of the specific animals to be treated, susae t i bi 1? > -larl from the infecting organism to the antibiotic and the stage and severity of the infection. Gener ally, the dosage is n th i m ad t m e rad y. * 1.0 m Approximately 25 m »j / l < and body weight per day, in divided dosages, the specific dosage .4 Stay with the doctor's profession. By dealing with some lenses with taxation and invention, it is possible to include other ingredients that are important to act and act. > s in the dosing unit. The MICRO-ORGANISM The methyl-bromide to obtain the compounds of this invention is a mutant strain of the onospora Carbonacea v Africana in accordance with that set forth in US Pat. No. 4,597,968, which is incorporated herein by reference. reference. how do you get this >; epa mutaute is presented in this application. The mutant strain of Mi c ro onospora Cartionacea VA V Africana was prepared in accordance with 3 or submitted to incubation. I m i l ent, 3 to mother stock SCC 1413 was submitted to thesis and thesis cor »N-m I, ro - =» oyuan? D? na (NTG) which resulted in the death of the As of 90 * 4 of the crop. One thousand five hundred isolates on these were those inalaros for hological activity? > a increased against S. aureus and E "cali. Isolated from single colony germinated in test tubes containing 10 ml. of germination medium and agiods at 25o r.p.m. on a rotating agitator at a temperature of 3 ° C for 48 hours. Fermentation studies were initiated by transferring 2.5 mL of the seed to Erlemneyer flasks of 250 ml. which contained r »l of medium axis fermentation and m < placed at 30 * C for 96 hours at 25o r.p.m. on a rotating agitator. SCC 1631 co or an improved productc.tr of the 13-384 complex was identified based on its improved tyioa tivity against S. aureus and E. coli. Strain SCC 1"'56 was isolated by mutation of NTG of SCC1631 followed by selection of the isolates on agar plates which contained 150 μg / mL of everninomic i na (complex of nitro and nitroso analogs). The SCC 21 6 strain was obtained by NTG from SCC 3756. The ceptum was used to isolate the mu lagéru strains of NCC of SCC 16.31 in In, high levels of Evernino ii na (complex of nitro analogues and nitroso), the protocols for both muta- tion studies were based on what was previously described.For the last or: - »study» of mutation, fermentation cations were extracted with ethyl acetate and the concentrates were chroma togra f side-, in lcs of thin capture Whatman Ll'RDF in a solvent system that 'insisted methanol chloroform methanol (9 *: 1) follower by bi oantogra f í a against S. aureus and E. coli to confirm the production of all the components of the antibiotic complex. everumomi ci na (nitrous and nitroso analogue), plates were screened by the use of the Shima au CS-930 TIC plate scanner and the extracts were harvested using HPLC. 3OÍ were defined, combined titles such as the sum e evernino i i na, analogous neither ro and noroso.

Claims (4)

1. A compound of the formula everni nomi ina - H OH 0H, H OMe Cl Me Compound A Cl. OH 0H, H OMe Cl Me Compue to B H OH = 0 OMe Cl Me Compue to D H OMe 0H, H OMe Cl. Me Compound E H OH 0H, H OH H Me Compue to F H OH 0H, H OMe H Me Compound G H OH 0H, H OMe Cl H or a pharmacologically acceptable solution.

2. A compound A according to claim 1, wherein Pl is Cl, P2 is OH, R3 is OH, P3 'is H, P4 is OMe, P5 is Cl and R is cies. Or a pharmaceutical s 3? > ament acceptable of the same. 3. A compound B according to claim 3 wherein Pl s H, P2 is OH, P3 is P3 'together are = 0, P4 is OMe, P is Cl and P6 is Me; or a s f ruméuicamente acepttable of the same. 4. Compound D according to claim 1 wherein R 1 is H, R 2 is OMe, r 3 is OH, P 3 'is H, R 4 is OH, P 5 is H and P 6 is Me 5 or a pharmaceutically salt. acceptable of it. 5. A compound F ci conformance with reaction 1 where Pl is H, P2 is OH, P3 is OH, R3 'is H, P4 is OMe, P5 is Cl and R6 ^ > Me; or a pharmaceutically acceptable salt thereof. 6. A • .F. F in accordance with the law. ac 1 on 1 where Pl is H, R2 is OH, R3 is OH, P 'is H, P4 is OMe, R5 is H and R6 s H; or an acceptable pharmaceutically acceptable salt thereof. 7. Compound G in accordance with rei indication 1 where Rl e-, H, R2 is OH, R3 is OH, P3 'is H, R4 is OMe, P5 is Cl and P6 is H1¡ or a slf rmacéu I love the acceptable one. 8. The compound H in accordance with the rei indication 1 where R 1 is H, R 2 is OH, P 3 is OH, R 3 'is H, P 4 is OMe, R 5 is H and R 6 is Me; or a pharmaceutically acceptable salt thereof. 9. A compound of the formula where R7 = CH3; R8 = CH3: (Compound C) or R7 = C (0) H; R8 = H (Compound H) or a pharmaceutically acceptable salt thereof,

3. 0. The compound C according to claim 9 where R7 is CH3, and R8 is CH3; or a pharmaceutically acceptable s l thereof. 11. Compound H in accordance with the rei indication 9 where R7 is C (0) H and R.8 is H; or a salt. pharmaceutically acceptable thereof. 12. The compound of the formula or a pharmaceutically acceptable salt of. same. 13. A composition comprising a compound in accordance with the invention and a pharmaceutically acceptable carrier material. 1.

4. A composition comprising a compound in accordance with the reagent 9 and a material, a pharmaceutically acceptable carrier. ; n 1 »A method for treating bacterial infection in a mammal that makes 1 at administration to the mammal that requires a antimicrobial effective amount of a compound in accordance with claim 1. 16. A method for the treatment of bacterial infections in a mammal comprising administering ion to the mammal in need of a bacterially effective amount of a compound in accordance with claim 9.