the present invention is directed to a compound having the formula ## str2 ## wherein r is h seq id no : 1 or oh seq id no : 2 , and to a method of producing same . when r is h , the compound is hereinafter designated compound ia . when r is oh the compound is hereinafter designated compound ib . the structure of the compounds have been determined by detailed analyses of spectral characteristics . electron impact ( ei , 70 ev ) and low resolution fast atom bombardment ( fab ; ms and ms - ms ) mass spectral data were obtained on a finnigan - mat tsq70b mass spectrometer . gc - ms analyses of the tms ( trimethylsilyl ) derivatives of total acid hydrolyzates 7ere performed on the same instrument . high resolution fab measurements were recorded on a finnigan - mat mat90 instrument . compound ia has the molecular formula c 50 h 80 n 8 o 17 [ from m + cs ]+: calcd 1064 . 5641 , found 1064 . 5585 ) by fab - ms . gc - ms analysis of the tms derivative of the total acid hydrolysate indicated approximately one equivalent each of threonine , 3 - hydroxyglutamic acid and 10 , 12 - dimethyltetradecanoic acid plus two equivalents of 4 - hydroxyproline . 1 h nmr spectrum : in cd 3 od at 400 mhz is seen in fig1 ; and 13 c nmr chemical shifts ( cd 3 od ): 11 . 6 , 19 . 7 , 20 . 2 , 20 . 7 , 27 . 0 , 28 . 1 , 30 . 3 ( 2 ×), 30 . 6 , 30 . 8 , 31 . 2 , 31 . 3 , 32 . 9 , 34 . 9 , 36 . 7 , 38 . 1 , 38 . 5 ( 2 ×), 39 . 4 , 45 . 9 , 51 . 2 , 56 . 1 , 56 . 3 , 57 . 1 , 57 . 9 , 58 . 3 , 60 . 7 , 62 . 4 , 68 . 2 , 70 . 6 , 70 . 9 , 71 . 0 , 71 . 3 , 73 . 8 , 75 . 8 , 76 . 9 , 116 . 2 ( 2 ×), 129 . 6 ( 2 ×), 133 . 0 , 158 . 5 , 169 . 2 , 172 . 5 , 172 . 9 , 173 . 4 , 174 . 5 , 174 . 6 , 175 . 7 , 177 . 3 ppm . compound ib has the molecular formula c 50 h 80 n 8 o 18 ( from [ m + cs ] + : calcd 1080 . 5590 , found 1080 . 5344 ) by fab - ms . gc - ms analysis of the tms derivative of the total acid hydrolyzate indicated approximately one equivalent each of threonine , 4 - hydroxyproline , 3 - hydroxyglutamic acid , 3 , 4 - dihydroxyproline , and 10 , 12 - dimethyltetradecanoic acid . fab - ms - ms of the [ m + h ] + ion indicates that ib contains 3 , 4 - dihydroxy - proline at the position preceeding the threonine residue . 1 h nmr spectrum in cd 3 od at 400 mhz is seen in fig2 ; and 13 c nmr chemical shifts ( cd 3 od ): 11 . 6 , 19 . 7 , 20 . 2 , 20 . 8 , 27 . 0 , 28 . 0 , 30 . 3 , 30 . 6 , 30 . 8 , 31 . 2 , 31 . 2 , 32 . 9 , 32 . 9 , 34 . 8 , 36 . 7 , 38 . 1 , 38 . 5 , 39 . 4 , 45 . 9 , 51 . 2 , 54 . 4 , 55 . 3 , 56 . 2 , 57 . 0 , 58 . 3 , 62 . 5 , 65 . 9 , 68 . 2 , 70 . 7 , 70 . 8 , 71 . 3 , 71 . 9 , 73 . 8 , 75 . 8 , 75 . 9 , 77 . 0 , 116 . 3 ( 2 c ), 129 . 7 ( 2 c ), 132 . 9 , 158 . 4 , 169 . 4 , 172 . 5 , 172 . 7 , 173 . 2 , 173 . 4 , 174 . 5 , 175 . 9 , 177 . 2 . on the basis of these and other data , compound i is believed with considerable certainty to have the structure indicated . compounds ia and ib are white solids , soluble in organic solvents such as methanol , ethanol , dimethylformamide , dimethyl sulfoxide , ethyl acetate and the like . compound i ( ia and ib ) has antifungal properties against both filamentous fungi and yeasts . it is particularly useful against organisms causing pathogenic mycotic infections such as candida albicans , candida tropicalis , candida pseudotropicalis , candida parapsilosis and the like . moreover , unlike a number of antifungal agents , such as amphotericin b , which while active against candida albicans and other fungal pathogens are limited in their ability because of the untoward and dangerous side effects , the antifungal agent of the present invention is not only a very effective but is substantially free of undesirable side reactions . red blood cell lysis , a harmful and potentially fatal side reaction is shown by many compounds at concentrations approaching the therapeutic dose and this property has limited the applicability of these compounds as drugs . the compound of the present invention would require a concentration of drug far above that required for therapeutic use before red blood cell lysis could occur . compound i also may be employed against filamentous fungi such as aspergillus species , penicillium species , fusarium species , alternaria species , neurospora species and the like . compound i also may be employed for the treatment of pneumocystis carinii , the causative agent of a pneumonia which is of particular severity to immune compromised patients such as those with acquired immune deficiency syndrome ( aids ). compound i is conveniently produced by cultivating a mutagenized form of zalerion arboricola produced as hereinafter described , and maintained in the merck culture collection as mf5533 . mf5533 has been deposited under the budapest treaty in the culture collection of the american type culture collection at 12301 parklawn drive , rockville , md . 20852 , and has been assigned accession number atcc 74030 . zalerion arboricola atcc 74030 is a mutagenized form of zalerion arboricola atcc 20957 which in turn is a mutagenized form of atcc 20868 . this mutant may be produced by cultivating a frozen vegetative mycelia of z . arboricola atcc 20957 with a mutagen , followed by plating , incubating and isolating as hereinafter more fully described . the major product in the cultivation of z . arboricola mf5533 is compound x seq id no : 3 represented by the formula ## str3 ## for the production of this mutant , a number of agents commonly used to produce mutants may be employed such as ultraviolet radiation , chemical mutagen , or intercalating agent . suitable chemical mutagens include n - nitroso - n - methylurethane and n - methyl - n &# 39 ;- nitro - n - nitrosoguanidine . in the present instance the z . arboricola mutant mf5533 , atcc 74030 was obtained by inoculating a frozen vegetative mycelium of z . arboricola mf5404 , atcc 20957 , disclosed and claimed in copending application ser . no . 07 / 878 , 137 , continuation application 492 , 024 , now abandoned into a seed medium , adding n - methyl - n &# 39 ;- nitro - n - nitrosoguanidine and cultivating , thereafter plating a portion of the growth on potato dextrose agar and incubating to develop colonies , and then transferring the separate colonies to slants of potato dextrose agar and incubating for 14 days at 25 ° c . to obtain cultures of mutants of z . arboricola , one of which was designated as 47 - 19 , and subsequently maintained in the merck culture collection as mf5533 . the colonial and morphological description of z . arboricola mf5533 atcc 74030 are as follows : colonies on potato - dextrose agar ( difco ) at 20 ° c . are slow - growing , attaining a diameter of 8 - 12 mm in one week . mature colonies ( 3 - 4 weeks ) on potato - dextrose agar effuse , with submerged and aerial hyphae , surface hairly , lanose , or funiculose , dull to moderately shiny , forming raised , densely compact colonies , with a substromatic texture due to dense condidia formation . colony color pale olive - brown , olive , olive - brown , finally olive - black , isabella color , sayal brown , tawny - olive , saccardo &# 39 ; s umber , sepia , brownish olive , raw umber , dark olive , olivaceous black ( capitalized color names from r . ridgway . 1912 . color standards and nomenclature , washington , d . c .). same colors in colony reverse . odor , exudates , and soluble pigments absent . hyphae ( in 3 % koh ) pale yellow - brown to olive - brown , septate , branched , often with irregular lateral or terminal lobes , 1 - 3 um wide , thin - to slightly thick - walled , with walls smooth to slightly incrusted or verrucose . aerial hyphae often adhering together in facicles . setae and hyphopodia absent . conidiogeneous cells monoblastic , scattered to dense , integrated , terminal and intercalary , arising directly from undifferentiated hyphae , at right to slightly acute angles . conidia originating as irregular chains filaments , or coils , later developing as compact , irregular masses of 6 - 25 cells . individual condial cells , 3 - 6 um in diameter , globose , subglobose , or slightly irregular to lobed , smooth to finely verruculose , yellow - brown to olive brown . compound i may be obtained by cultivating z . arboricola mf5533 in a suitable nutrient medium under conditions hereinafter described until a substantial amount of antifungal activity can be detected in the culture medium , harvesting by extracting the active components from the fermentation medium with a suitable solvent , concentrating the solution containing the desired component , then subjecting the concentrated material to chromatographic separation to isolate compound i from other metabolites also present in the cultivation medium . a suitable nutrient medium for producing compound i is one containing sources of carbon and nitrogen assimilable by the microorganism and also containing low levels of inorganic salts . the medium may be supplemented with trace metals , although if complex sources of carbon and nitrogen are employed , the trace metals are usually present in the complex sources . the sources of carbon include glycerol , sugars , sugar alcohols , starches and other carbohydrates , or carbohydrate derivatives such as dextran , cerelose , as well as complex nutrients such as oat flour , corn meal , millet , corn and the like . the exact quantity of the carbon source which is utilized in the medium will depend , in part , upon the other ingredients in the medium , but it is usually found that an amount of carbohydrate between 0 . 5 and 40 percent by weight of the medium is satisfactory . these carbon sources can be used individually or several such carbon sources may be combined in the same medium . the sources of nitrogen include amino acids such as glycine , arginine , threonine , methionine and the like , ammonium salt , and complex sources such as yeast hydrolysates , yeast autolysates , yeast cells , tomato paste , soybean meal , casein hydrolysates , yeast extracts , corn steep liquors , distillers solubles , cottonseed meal , meat extract , and the like . the various sources of nitrogen can be used alone or in combination in amounts ranging from 0 . 2 to 10 percent by weight of the medium . among the nutrient inorganic salts , which can be incorporated in the culture media are the customary salts capable of yielding sodium , potassium , magnesium , calcium , phosphate , sulfate , chloride , carbonate , and like ions . also included are trace metals such as cobalt , manganese , iron , molybdenum , zinc , cadmium , and the like . although the growth medium may be prepared in a conventional manner from the foregoing nutrients , the presence of certain nutrients and / or combination of nutrients favor the production of compound i . thus , ammonium salts are important as an immediate source of nitrogen and monobasic potassium phosphate is important for ph control . mannitol is especially useful in compositions , not only for enhancing the amount of desired product formed but also in improving the rate of production of the desired product . the cultivation medium may be either liquid or solid . representatives suitable media for production of compound i are the following : ______________________________________tg106 medium per literd - mannitol 100 gnz - amine type e * 33 gfidco 8005 yeast extract 10 g ( nh . sub . 4 ). sub . 2 so . sub . 4 5 gkh . sub . 2 po . sub . 4 9 gp - 2000 2 ml______________________________________ * casein hydrolysate , sheffield products , kraft , inc . sp - 5 meduim per litermannitol 80 gkh . sub . 2 po . sub . 4 9 gcerelose 10 gpharmamedia * 20 gpresterile ph 7 . 3______________________________________ * yellow flour from embryo of cottonseed containing nonhydrolyzed globular protein . traders protein , buckeye oilseed products co ., memphis tenn . ______________________________________rg2 medium rg120 medium per liter per litermannitol 44 g mannitol 91 gcorn steep liquor 4 g corn steep liquor 4 mllard water 4 g lard water 4 gpectin 10 g pectin 10 gkh . sub . 2 po . sub . 4 2 g kh . sub . 2 po . sub . 4 2 gtomato paste 4 g tomato paste 4 gpeptonized milk 4 g peptonized milk 4 gglycine 2 g glycine 2 gpeanut meal 4 g peanut meal 4 gph adjusted to 7 . 0 ph adjusted to 7 . 0tg102 medium tg103 medium per liter per literd - mannitol 40 g d - mannitol 40 gbacto peptone * 33 g bacto - peptone * 33 gbacto - yeast extract 10 g bacto - yeast extract 10 g ( nh . sub . 4 ). sub . 2 so . sub . 4 5 g ( nh . sub . 4 ). sub . 2 so . sub . 4 5 gkh . sub . 2 po . sub . 4 9 g kh . sub . 2 po . sub . 4 9 gno ph adjustment no ph adjustments2 medium s6 medium per liter per literd - mannitol 44 g d - mannitol 44 gkh . sub . 2 po . sub . 4 2 g kh . sub . 2 po . sub . 4 2 gglycine 2 g glycine 2 gpeptonized milk 15 g peptonized milk 15 glactic acid 2 g lactic acid 2 gtrace elements 10 ml trace elements 10 ml soybean oil 10 gph 7 . 0 ph 7 . 0 ( pre - sterilization ) ( pre - sterilization ) f204 solid medium per 250 - ml flask base liquid per litermillet 15 g ardamine ph (**) 33 . 0 gbase liquid 15 ml sodium tartrate 6 . 6 g feso . sub . 4 . 7h . sub . 2 o 0 . 66 g monosodium glutamate 6 . 6 ml corn oil 6 . 6 ml no ph adjustment______________________________________ ** yeast autolysate , yeast products inc . clifton , new jerseyf4 - sf solid medium per 250 - ml flask base liquid per litercracked corn 15 g ardamine ph 0 . 2 gbase liquid 10 ml h . sub . 2 po . sub . 4 0 . 1 g mgso . sub . 4 . 7h . sub . 2 o 0 . 1 g sodium tartrate 0 . 1 g feso . sub . 4 . 7h . sub . 2 o 0 . 01 g znso . sub . 4 . 7h . sub . 2 o 0 . 01 g no ph adjustment______________________________________ production of the desired compound using one of the foregoing or similar medium , is usually initiated by first inoculating a nutrient seed medium with a frozen vegetative mycelia of z . arboricola mf 5533 , and the inoculated medium incubated for at least 3 days to produce a broth containing organisms which serves as seed in the production of the compounds of formula ( i ). instead of using all or an aliquot of the fermentation broth for production , an aliquot of the broth may be employed in a second stage production of seed medium . depending on the size of the production contemplated , several stages of seed medium production may be carried out prior to utilization of the fermentation broth as seed in the ultimate production of compound i . the seed medium is generally in the ph range of 5 to 8 . 1 , optimally 6 to 7 . 5 . ______________________________________ per / liter______________________________________corn steep liquor 5 gd - mannitol 25 gglucose monohydrate 10 gpharmamedia 20 gkh . sub . 2 po . sub . 4 9 gfeso . sub . 4 . 7h . sub . 2 o 10 mgmnso . sub . 4 . 4h . sub . 2 o 10 mgcucl . sub . 2 . 2h . sub . 2 o 0 . 25 mgcacl . sub . 2 . 2h . sub . 2 o 1 mgh . sub . 3 bo . sub . 3 0 . 56 mg ( nh . sub . 4 ). sub . 6 mo . sub . 7 o . sub . 24 . h . sub . 2 o 0 . 19 mgznso . sub . 4 . 7h . sub . 2 o 2 mg______________________________________ ______________________________________ per / liter______________________________________corn steep liquor 5 gtomato paste 40 goat flour 10 gglucose 10 gfeso . sub . 4 . 7h . sub . 2 o 10 mgmnso . sub . 4 . 4h . sub . 2 o 10 mgcucl . sub . 2 . 2h . sub . 2 o 0 . 25 mgcacl . sub . 2 . 2h . sub . 2 o 1 mgh . sub . 3 bo . sub . 3 0 . 56 mg ( nh . sub . 4 ). sub . 6 mo . sub . 7 o . sub . 24 . h . sub . 2 o 0 . 19 mgznso . sub . 4 . 7h . sub . 2 o 2 mg______________________________________ in carrying out the process , a slant section of a preserved culture of mf5533 atcc 20958 is inoculated into an appropriate seed medium and the flasks incubated with or without agitation at temperatures in the range of from about 15 ° c . to about 30 ° c . for from 2 to 30 days , preferably 20 ° to 28 ° c . for 2 to 14 days . agitation when employed is preferably in the range of from 150 to 220 rpm but may be up to 400 rpm . when growth is abundant , usually between 2 and 5 days , the growth may be used to inoculate the production medium for the production of the compounds of this invention . preferably however , a second stage fermentation and , frequently a third or fourth stage fermentation are carried out by inoculating with a portion of the culture growth and then employing similar conditions and incubation time of from about 1 to 6 days . the growth then is employed to inoculate the production medium . the fermentation production medium inoculated with the culture growth is incubated for 3 to 30 days , usually 7 to 14 days , with or without agitation . the fermentation may be conducted aerobically at temperatures ranging from about 20 ° c . to about 40 ° c . for optimum results , it is most convenient to conduct these fermentations at a temperature in the range of from about 24 ° c . to about 30 ° c . temperatures of about 24 °- 28 ° c . are most preferred . the ph of the nutrient medium suitable for producing the instant compounds can vary from about 5 . 0 to 8 . 5 with a preferred range of from about 5 . 5 to 6 . 0 . after the appropriate period for the production of the desired compound or compounds , the latter is recovered from the fermentation medium as hereinafter more fully described . after completion of the cultivation , compound i is harvested and isolated from the medium . the exact steps may vary somewhat on whether the fermentation is carried out in liquid or solid medium . when the fermentation is carried out on a solid medium , the first step may be adding an alcoholic solvent to the fermentation medium , thoroughly mixing , then filtering , recovering and concentrating the aqueous alcohol filtrate . the concentrated filtrate may be first back - extracted or washed with a lower aliphatic hydrocarbon solvent such as hexane or other alkane to remove alkane soluble impurities . when the fermentation is carried out in a liquid medium , in one method , the mycelial solids may be separated by filtration or centrifugation and recovered from the fermentation medium . alcohol is added to the mycelial cake , and the mycelial solid thoroughly mixed with a lower alkanol to extract the desired product and the mixture filtered or centrifuged , and the filtrate or supernatant collected and concentrated . in an alternative method , the whole broth can be extracted by the addition of one volume of lower alkanol , preferably methanol , and filtered or centrifuged to remove solid impurities . the lower alkanol suitable for extracting the active agent from the solid nutrient medium or the mycelial pad obtained on centrifugation or filtration include methanol , ethanol , isopropanol or higher alkanols . methanol is preferred . the alkanol extract from either separation is then placed onto a column for chromatographic separation steps . adsorbents available commercially such as styrenedivinylbenzene copolymers available commercially as &# 34 ; diaion &# 34 ; hp - 20 , hp - 30 , hp - 40 , sp - 207 ( mitsubishi chemical industries , ltd .) and &# 34 ; amberlite &# 34 ; xad - 2 , xad - 4 , xad - 16 ( rohm and haas co . ), may be employed for the initial isolations . in carrying out the separation steps , the composition of the alkanol extract is adjusted to 50 percent water and adsorbed on the hp - 20 or other selected resin and then eluted with 100 percent alkanol , preferably methanol . conventional column chromatography may be employed . when conventional chromatographic separation is employed , &# 34 ; sephadex &# 34 ; lh - 20 ( pharmacia ) or silica gel may be employed although silica gel is preferred . in the fractionation and recovery of the active component by chromatography on silica gel , ester / alcohol / water or dichloromethane / alcohol / water may be employed to provide good separations . a mixture of ethyl acetate , methanol , and water or 5 percent aqueous acetic acid has been found to be especially useful . when a dextran adsorbent such as &# 34 ; sephadex &# 34 ; lh - 20 , is employed , a chlorohydrocarbon / hydrocarbon / alcohol solvent system may be employed . a mixture of methylene chloride / hexane / methanol has been found to be especially useful . in carrying out the hplc separation , the alcohol solution containing material recovered from the conventional chromatography is concentrated and the residue dissolved in methylene chloride / methanol / water or ethyl acetate / methanol / water in the same ratio as found in the mobile phase and placed on a column packed with commercial silica gel resin and eluted at about 800 - 2000 psi which produces a flow rate of about 10 ml / min . the separation is monitored by uv at 276 nm . compound i is active against many fungi , and also against pneumocystis carinii . the antifungal properties may be illustrated with the minimum fungicidal concentration ( mfc ) determinations against certain candida organisms in a microbroth dilution assay carried out in yeast nitrogen base ( difco ) with 1 percent dextrose ( ynbd ). in carrying out the assay , compound i was solubilized in 10 percent dimethyl sulfoxide ( dmso ) and diluted to 2560 μ / ml . the compound was then diluted to 256 μg / ml in ynbd . 0 . 15 ml of the suspension was dispensed to the top row of a 96 - well plate ( each well containing 0 . 15 ml of yndb ) resulting in a drug concentration of 128 μg / ml . two - fold dilutions were then made from the top row to obtain final drug concentrations ranging from 128 to 0 . 06 μg / ml . the yeast cultures , maintained on sabouraud dextrose agar were transferred to ym broth ( difco ) and incubated overnight at 35 ° c . with shaking ( 250 rpm ). after incubation , each culture was diluted in sterile water to yield a final concentration of 1 - 5 × 10 6 colony forming units ( cfu )/ ml . 96 - well microplates were inoculated using a mic - 2000 ( dynatech ) which delivers 1 . 5 μl per well yielding a final inoculum per well of 1 . 5 - 7 . 5 × 10 3 cells . the microplates were incubated at 35 ° c . for 24 hours . the minimum inhibitory concentrations ( mics ) were recorded as the lowest concentrations of drug showing no visible growth . after recording the mic , the plates were shaken to resuspend the cells . thereafter , 1 . 5 μl samples from the wells in the 96 - well microplate were transferred to a single well tray containing sabouraud dextrose agar . the inoculated trays were incubated 24 hours at 28 ° c . and then read . the mfc is defined as the lowest concentration of drug showing no growth or less than 4 colonies per spot . the results ( three samples ) are seen in the following table : ______________________________________ minimum fungicidal concentration ( μg / ml ) fungi compound compoundstrain no . ia ib______________________________________candida albicansmy 1055 2 1my 1028 2 0 . 5my 1750 4 1candida tropicalismy 1012 1 0 . 12candida pseudotropicalismy 1100 4 2______________________________________ compound i is useful for inhibiting or alleviating pneumocystis carinii infections . in a representative study , the effectiveness of compound i in rats were determined . sprague - dawley rats ( weighing approximately 250 g ) were immunosuppressed with dexasone in the drinking water ( 2 mg / ml ) and maintained on a low protein diet for 5 weeks to induce the development of pneumocystis pneumonia from a latent infection . before drug treatment 2 rats were sacrificed to confirm the presence of pneumocystis carinii pneumonia ( pcp ); both rats had infections . the remaining rats ( weighing approximately 150 g ) were distributed into groups of 6 and injected twice daily for four days subcutaneously with compound in 0 . 25 ml of vehicle ( 10 % dmso in water or water ). the control group of 5 rats received vehicle alone . all animals continued to receive dexasone in the drinking water and low protein diet during the drug treatment period . at the completion of treatment all animals were sacrificed , the lungs were removed and processed , and the extent of disease determined by microscopic analysis of stained slides . the results of this study are shown as the log means number of cysts per animal lung as determined by examining 20 - 1000 × microscopic fields ( as seen in the following table ) and the standard of error of the geometric mean . all groups were compared using the student t - test and results marked (*) are significant . table______________________________________ log mean # cysts reduction (± segm ) of cysts survivors______________________________________dmso control 7 . 26 ± . 12 -- 6 / 6compound ia1 . 20 mg / kg 5 . 58 ± . 11 * 97 . 9 % 5 / 60 . 60 mg / kg 5 . 68 ± . 19 * 97 . 4 % 6 / 60 . 30 mg / kg 5 . 99 ± . 18 * 94 . 6 % 5 / 60 . 15 mg / kg 6 . 32 ± . 17 * 88 . 5 % 6 / 6compound ib0 . 60 mg / kg 5 . 41 ± . 17 * 99 . 1 % 6 / 60 . 30 mg / kg 5 . 22 ± . 10 * 99 . 4 % 6 / 60 . 15 mg / kg 5 . 59 ± . 11 * 98 . 7 % 6 / 60 . 07 mg / kg 5 . 97 ± . 18 * 96 . 8 % 6 / 60 . 03 mg / kg 6 . 16 ± . 14 * 95 . 0 % 6 / 6______________________________________ the outstanding properties are most effectively utilized when the compound is formulated into novel pharmaceutical compositions with a pharmaceutically acceptable carrier according to conventional pharmaceutical compounding techniques . the novel compositions contain at least a therapeutic antifungal or antipneumocystis amount of the active compound . generally , the composition contains at least 1 percent of weight of compound i . concentrate compositions suitable for dilutions prior to use may contain 90 percent or more by weight . the compositions include compositions suitable for oral , rectal , topical , parenteral ( including subcutaneous , intramuscular , and intravenous ), pulmonary ( nasal or buccal inhalation ), nasal administration , or insufflation . the compositions may be prepacked by intimately mixing compound i with the components suitable for the medium desired . when the compound is for antifungal use any method of administration may be used . for treating mycotic infection oral administration is frequently preferred . when oral administration is to be employed , it may be with a liquid composition . for liquid preparations , the therapeutic agent is formulated with liquid carriers such as water , glycols , oils , alcohols , and the like , and for solid preparations such as capsules and tablets , solid carriers such as starches , sugars , kaolin , ethyl cellulose , calcium and sodium carbonate , calcium phosphate , kaolin , talc , lactose , generally with lubricant such as calcium stearate , together with binders , disintegrating agents and the like . because of their ease in administration , tablets and capsules represent the most advantageous oral dosage form . it is especially advantageous to formulate the compositions in unit dosage form ( as hereinafter defined ) for ease of administration and uniformity of dosage . composition in unit dosage form constitutes an aspect of the present invention . the compound i also may be formulated in therapeutic compositions for intravenous or intraperitoneal injection and may be presented in unit dosage form in ampoules or in multidose containers , if necessary with an added preservative . the compositions may also take such forms as suspensions , solutions or emulsions in oily or aqueous vehicles such as 0 . 85 percent sodium chloride or 5 percent dextrose in water , and may contain formulating agents such as suspending , stabilizing and / or dispersing agents . buffering agents as well as additives such as saline or glucose may be added to make the solutions isotonic . the drug also may be solubilized in alcohol / propylene glycol or polyethyleglycol for drip intravenous administration . alternatively , the active ingredients may be in powder form for reconstituting with a suitable vehicle prior to administration . the term &# 34 ; unit dosage form &# 34 ; as used in the specification and claims refer to physically discrete units , each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the pharmaceutical carrier . examples of such unit dosage forms are tablets , capsules , pills , powder packets , wafers , measured units in ampoules or in multidose containers and the like . a unit dosage of the present invention will generally contain from 100 to 200 milligrams of one of the compounds . when the compound is to be employed for control of pneumocystis infections it is desirable to directly treat lung and bronchi . for this reason , inhalation methods are preferred . for administration by inhalation , the compounds of the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs of nebulisers . the compounds may also be delivered as powders which may be formulated and the powder composition may be inhaled with the aid of an insufflation powder inhaler device . the preferred delivery system for inhalation is a metered dose inhalation ( mdi ) aerosol , which may be formulated as a suspension or solution of compound i in suitable propellants , such as fluorocarbons or hydrocarbons . another method of administration is insufflation , particularly in the infection has spread to the ears and other body cavities . if the application is to be topical , the drug may be formulated in conventional creams and ointments such as white petrolatum , anhydrous lanolin , cetyl alcohol , cold cream , glyceryl monostearate , rose water and the like . usually a 1 to 2 percent cream solution is prepared and applied to the area to be treated . the following examples illustrate the invention but are not to be construed as limiting . a frozen vegetative mycelium of z . arboricola mf5404 atcc 20957 was inoculated into 50 milliliters of kf seed medium contained in a 250 milliliter erlenmeyer flask and n - methyl - n &# 39 ;- nitro - n - nitrosoguanidine ( ntg ) was added to a final concentration of 7 . 5 micrograms per milliliter . the flask was shaken at 220 rpm at 25 ° c . for five days to obtain broth containing cells grown in the presence of ntg . a portion of the broth was plated on the surface of potato dextrose agar and the plates incubated for 14 days at 25 ° c . to obtain spores of the microorganism . the spores were harvested , diluted in sterile saline and plated on the surface of potato dextrose agar and incubated at 25 ° c . for 7 days for colony formation . the colonies were isolated by transferring each colony to separate slants of potato dextrose agar . the inoculated slants were incubated at 25 ° c . for 14 days and a plug from the slants separately taken and inoculated into 20 milliliters of kf medium to produce seeds , then , 2 milliliters of the seed used to inoculate 40 milliliters of sp - 5 medium which was then incubated at 25 ° c . for 14 days . the broth then was extracted with methanol and the extract tested for the production of compound x and other components by hplc . one of the slants designated culture 47 - 19 was used in the production hereinafter described . culture 47 - 19 was subsequently re - isolated and preserved in the merck culture collection as mf 5533 . seed cultures were first prepared in several stages . as an initial step , 54 milliliters of p34 - 2 medium was inoculated with a plug from a frozen vial of zalerion arboricola tentatively designated culture 47 - 19 and subsequently re - isolated and preserved in the merck culture collection as mf5533 . the inoculated medium was incubated with shaking at 220 rpm at 25 ° c . for four days . a twenty - milliliter sample of this seed medium was used to inoculate each of four 2 - liter flasks containing 500 milliliters of p34 - 2 medium and the inoculated medium incubated at 25 ° c . for four days at 220 rpm . the flask contents were then pooled and used to inoculate a 300 - liter seed fermenter containing 180 liters of p34 - 2 medium and 2 milliliters / liter of polypropylene glycol p - 2000 to reduce foaming . the fermenter was operated for six days at a temperature of 25 ° c ., an air flow of 90 liters per minutes , a pressure of 0 . 7 kg / cm 2 gauge , and an agitator speed of 200 rpm . a 25 liter sample of this seed then was used to inoculate an 800 liter seed fermenter containing 475 liters of p34 - 2 medium and 2 milliliter / liter of p - 2000 and cultivated for four days at 25 ° c ., air flow of 250 liter / minute , a pressure of 0 . 7 kg / cm 2 gauge and agitator speed of 150 rpm . 425 liters of the seed broth thus prepared was inoculated into 13 , 700 liters of tg 106 medium in a 19 , 000 liter production fermenter . fermentation of the mixture was carried out at a temperature of 25 ° c ., air flow of 6300 liters / minute , a pressure of 0 . 7 kg / cm 2 gauge and agitator speed of 80 rpm . the ph was allowed to decrease from an initial value of 6 . 0 to 5 . 5 and then maintained at 5 . 5 ± 0 . 2 using sodium hydroxide or sulfuric acid . the cultivation was continued for twelve days , after which time the broth was harvested for product isolation . 3100 gallons of whole broth was first extracted with 1900 gallons of methanol . the methanol was clarified by centrifugation to obtain 4250 gallons of clarified liquid as first extract and 650 gallons of solid . the latter was re - extracted with 1500 gallons of methanol and centrifuged to obtain 1550 gallons of clarified liquid as second extract . the solids from this centrifugation were extracted with 1500 gallons of 80 percent aqueous methanol and centrifuged to obtain a third extract . the first extract was loaded on an sp - 207 ( brominated polystyrene - divinylbenzene copolymer ) adsorption column and washed with 65 % aqueous methanol . some product breakthrough was noted in the 65 percent methanol wash . the 65 percent methanol wash and selected side outs from the first sp - 207 column , amounting to 900 gallons , were combined with the second and third extracts from the whole broth and combined extracts totalling 4050 gallons were absorbed on an sp - 207 column and eluted with 100 percent methanol . the rich cut eluate from the first sp - 207 column was diluted , adsorbed and eluted with 100 percent methanol from an hp - 20 columns . the rich cut eluate from the second sp - 207 column was combined , diluted with water , adsorbed to two hp - 20 columns , and eluted with 100 percent methanol . the three hp - 20 rich cut eluates were combined , diluted with water and then adsorbed and eluted with 100 percent methanol from an sp - 207 column for concentration and dewatering . the rich cut was concentrated 4 × by distillation . to a 500 ml aliqnot of the 4 × concentrate from the foregoing columns was added 4 . 5 liters of isopropyl acetate to separate the product which precipitates from p - 2000 which remains in solution . the precipitate was then prepared for preparative hplc to be carried out on a commercial column identified as dorr - oliver peak performer equipped with a prochrom lc 150 ve 15 cm . column charged with 3 kilograms of ` matrex ` irregular silica gel ( amicon ) of 20 μm particle size , 60 å pore size . in the preparation the precipitate was first filtered , vacuum dried , and dissolved in 550 milliliters of 2 : 2 : 1 ethyl acetate / methanol / 5 % acetic acid solvent mixture . the solvent composition was then adjusted to the feed composition of 76 / 16 / 8 ethyl acetate / methanol / 5 % aqueous acetic acid and 715 milliliters of the mixture was immediately injected onto the prochram lc 150 ve column at a flow rate of 605 ml / min . using the mobile phase of 80 / 10 / 5 ethyl acetate / methanol / 5 % aqueous acetic acid and twenty - two 2 - liter fractions collected . fraction 16 was concentrated to dryness and 100 milligrams of it was reconstituted in 2 milliliters of 80 : 20 : 2 methylene chloride / methanol / water . the solution was filtered to clarify it and the clear solution injected onto a 50 centimeter × 22 . 5 millimeter whatman silica gel hplc column . the mobile phase for the chromatography was 80 : 20 : 2 methylene chloride / methanol / water and the flow rate was 10 milliliters / minute . the chromatography was monitored using uv detector at 276 nm and also by analytical hplc of the fractions . both detection methods indicated a baseline separation of compound ia , ib and x , all of which were present in the original mixture . a total of nine identical injections were performed in the manner above indicated and the factions containing compound ia were combined , concentrated to dryness and reconstituted into 100 milliliters of 50 : 50 methanol / water . this solution was adsorbed to 20 milliliters of hp - 20 , eluted with methanol and dried under vacuum to obtain 91 milligrams of compound ia . the remaining 4 × concentrate from the third sp - 207 column in example 3 was added to 10 volumes of isopropyl acetate to precipitate compound x , ia and ib . the precipitate was collected by centrifugation and the material chromatographed on four 500 ml silica gel columns eluted with 85 : 10 : 5 ethyl acetate / methanol / 5 % aqueous acetic acid . a composite sample from one of the silica columns containing 60 g of compound ib and 20 g of compound ia ( as determined by hplc ) was chromatographed on a 150 liter silica column run in a similar manner . four 50 gallon fractions were collected followed by twenty - three 15 gallon fractions . one of the 15 gallon fractions rich in compound ib (& gt ; 95 % as determined by hplc ) was concentrated to dryness , reconstituted in ethyl acetate / methanol / water ( 76 / 16 / 8 ) and further purified on a 1 liter silica column , eluted with 85 / 10 / 5 ethyl acetate / methanol / 5 % aqueous acetic acid . the rich cuts were combined , concentrated , redissolved in 50 / 50 meoh / h 2 o and desilicated on a 40 ml hp - 20 column . compound ib was eluted with 100 % meoh . the cuts rich in compound ib were combined , subjected to a rotary evaporator to vaporize the methanol and obtain an aqueous solution which was lyophilized to obtain compound ib as a white powder . the white powder had the spectral characteristics set forth previously for compound ib . in a manner similar to that described in part a of example 1 , mutant mf 5533 was obtained from mf 5404 except that the seed from the seed flasks were used to inoculate 40 milliliters of tg - 106 medium and the latter incubated at 25 ° c . with shaking at 25 ° c . for 14 days . 1000 tablets , each containing 500 milligrams of compound ia are prepared from the following formulation : ______________________________________compound grams______________________________________compound ia 500starch 750dibasic calcium phoshate hydrous 5000calcium stearate 2 . 5______________________________________ the finely powered ingredients are mixed well and granulated with 10 percent starch paste . the granulation is dried and compressed into tablets . 1000 hard gelatin capsules , each containing 500 milligrams of compound ib are prepared from the following formulation : a uniform mixture of the ingredients is prepared by blending and used to fill two - piece hard gelatin capsules . 250 milliliters of an injectable suspension are prepared by conventional procedures having the following formulation : an ointment suitable for topical application may be prepared by intimately dispersing 13 milligrams of compound ia in 1 gram of commercially available polyethylene / hydrocarbon gel . ______________________________________ per canister______________________________________compound ib 24 mglecithin nf , liquid concentration 1 . 2 mgtrichlorofluoromethane 4 . 025 gdichlorodefluoromethane 12 . 15 g______________________________________ a culture of z . arboricola atcc 20868 was grown on potato dextrose agar in petri plates at 25 ° c . for 3 weeks . ten milliliters of 0 . 3m tris buffer , ph 7 , were added to the plates and the spores scraped off the surface into the buffer with a sterile cotton swab . the suspension in the buffer was decanted off and the procedure repeated twice . the spore suspensions were combined and filtered through glass wool to remove large clusters of spores . the suspension filtrate was centrifuged at first at 600 rpm then at 700 rpm and finally at 800 rpm , each time for 3 minutes with the pellet being discarded after each centrifugation . the supernatant liquid from the third centrifugation was then centrifuged at 3000 rpm for 5 minutes . the pellet from this centrifugation was resuspended in 3 milliliters of 0 . 3m tris buffer and used for mutagenic treatment . this suspension contained from 10 3 to 10 4 spores per milliliter . to the spore suspension was added 100 μg / ml of n - nitroso - n - methylurethane and the resulting mixture shaken at 300 rpm for 20 minutes at room temperature . at the end of this period , the mixture was centrifuged and the supernatant liquid was removed . the pellet was washed twice with 0 . 3m tris buffer ph 7 . 0 and then resuspended in the same buffer and after appropriate dilutions plated on potato dextrose agar for forming isolated colonies . the plates were incubated at 25 ° c . for two weeks for colony formation . the colonies were isolated by separately transferring to slants of potato dextrose agar . the inoculated slants were incubated at 25 ° c . for 10 - 14 days and a plug from the slants taken and tested for the production of compounds x and other components in the fermentation by hplc assay . a plug from one of the slants initially designated as z7 - 9 , was placed in the merck culture collection as mf 5404 and deposited with the american type culture collection as atcc 20957 . __________________________________________________________________________sequence listing ( 1 ) general information :( iii ) number of sequences : 3 ( 2 ) information for seq id no : 1 :( i ) sequence characteristics :( a ) length : 6 ( b ) type : amino acid ( c ) strandedness : na ( d ) topology : circular ( xi ) sequence description : seq id no : 1 : xaathr xaaxaaxaaxaa15 ( 2 ) information for seq id no : 2 :( i ) sequence characteristics :( a ) length : 6 ( b ) type : amino acid ( c ) strandedness : na ( d ) topology : circular ( xi ) sequence description : seq id no : 1 : xaathrxaaxaaxaa xaa15 ( 2 ) information for seq id no : 3 :( i ) sequence characteristics :( a ) length : 6 ( b ) type : amino acid ( c ) strandedness : na ( d ) topology : circular ( xi ) sequence description : seq id no : 1 : xaathrxaaxaaxaaxaa 15
a new antibiotic cyclic lipopeptide having the formula ## str1 ## wherein r is h seq id no : 1 or oh seq id no : 2 and a method of producing is described . the agent has very high activity against human pathogens and is of very low mammalian toxicity .