IE42455B1 - Alkylated derivatives of antibiotic bm123 - Google Patents
Alkylated derivatives of antibiotic bm123Info
- Publication number
- IE42455B1 IE42455B1 IE2651/75A IE265175A IE42455B1 IE 42455 B1 IE42455 B1 IE 42455B1 IE 2651/75 A IE2651/75 A IE 2651/75A IE 265175 A IE265175 A IE 265175A IE 42455 B1 IE42455 B1 IE 42455B1
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- lower alkyl
- phenyl
- trans
- substituted
- methanol
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
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- Genetics & Genomics (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Saccharide Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
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Abstract
Reductive alkylation with aldehydes R1-CHO or ketones R2-CO-R3 results in alkylation of the antibiotics BM 123 gamma 1, gamma 2 or mixtures thereof at the terminal primary amino group. With ketones there is single N-alkylation, while with aldehydes the primary amino group is substituted once or twice, depending on the amount of reagents, and the proximal secondary amino group on the side chain can likewise be alkylated. The meanings of R1, R2 and R3 are to be found in Claim 1. The resulting derivatives have a potent antibacterial action; they can be used for the treatment of infectious diseases.
Description
This invention relates to a novel group Of antibiotics and, more particularly, is concerned with a novel series of potent antibacterial agents derived by reductive alkylation of antibiotic BM123/^ with an aldehyde or ketone of the following general formulae:
R1-CHO
It r2-c-r3 wherein R^ is hydrogen, lower alkyl, halo substituted lower alkyl, lower alkenyl, phenyl, monosubstituted phenyl, phenyl lower alkyl, 2-furyl, methyl substituted 2-furyl, 2-thienyl, methyl substituted 2-thienyl, 2-pyrryl, methyl substituted 2-pyrryl, 2-pyridyl or 2-guinolyl; R2 is lower alkyl, halo substituted lower alkyl or phenyl lower alkyl; R3 is lower alkyl, halo
- 2 42455 substituted lower alkyl, lower alkenyl, lower eycloalkyl, phenyl, monosubstituted phenyl, phenyl lower alkyl or monosubstituted phenyl lower alkyl; and R2 and R3 taken together is tetramethylene, pentamethylene or hexamethylene. Suitable lower alkyl and halo substituted lower alkyl groups contemplated by the present invention are those having up to six carbon atoms wherein halo is exemplified by chloro, bromo, and iodo such as methyl, ethyl, isopropyl, sec-butyl, n-amyl, dichloromethyl, 2-bromoethyl, 2,3-diiodopropyl, /-chloropropyl, etc. Suitable lower alkenyl groups are those having up to four carbon atoms such as vinyl, allyl, propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, isobutenyl, etc. Suitable lower eycloalkyl groups are cyclopentyl, cyclohexyl, and cycloheptyl. Suitable monosubstituted phenyl groups contemplated by the present invention are, for example, p-acetamidophenyl, m-nitrophenyl, m-mercaptophenyl, o-anisyl, ja-anisyl, o-tolyl, g-tolyl, and the like whereas phenyl lower alkyl is exemplified by benzyl, α-phenylethyl, and β-phenylethyl. Suitable monosubstituted phenyl lower alkyl groups may be o, in, or £-chlorobenzyl, (g-aminophenyl)ethyl, β-(m-nitrophenyl)ethyl, etc. Suitable methyl substituted 2-furyl, 2-thienyl, and 2-pyrryl groups which may be employed are, for example, 5-methyl-2-furyl, 3,4-dimethyl-2-furyl, 4-methyl-2-thienyl, 3,5-dimethyl-2-thienyl, 5-methyl-2-pyrryl,
1,3,4-trimethyl-2-pyrryl, and the like.
The reductive alkylation process whereby the novel antibacterial agents of the present invention may be prepared is carried out as follows. Antibiotic
BM123V/, BM123or 814123/^ is dissolved in a suitable solvent such as water, methanol, methyl cellosolve, or mixtures thereof, an amount in excess of an equimolar amount of the desired aldehyde or ketone is then added followed by the addition of a reductive sufficiency of sodium cyanoborohydride. The pH of the reaction mixture is maintained at 6.0-8.0 with dilute mineral acid during the course of the reaction. After one to 24 hours at ambient temperature (10°-35°C.), the reaction mixture is evaporated to dryness in vacuo and the residue is triturated with methanol and filtered. The fii fcrate is diluted with acetone and the solid product that precipitates is removed by filtration and dried in vacuo.
Aldehydes which may be so employed in the above process are, for example, acetaldehyde, propionaldehyde, butyraldehyde, isobutyraldehyde, crotonaldehyde, valeraldehyde, benzaldehyde, p-cyanobenzaldehyde, salicylaldehyde, cinnamaldehyde, trichloroacetaldehyde, etc. Ketones which may be so employed in the above process are, for example, acetone, 2-butanone, 1,3-dibromoacetone, chloroacetone, acetophenone, m-chloroacetophenone, £-bromoacetophenone, £-trifluoromethylacetophenone, m-nitroacetophenone, £-dimethylaminoacetophenone, etc.
The products are obtained from the reductive alkylation reaction mixtures by standard procedures such as precipitation, concentration, solvent extraction or combinations of these procedures. After isolation, the products may be purified by any of the generally known methods for purification. These include recrystalliza30 tion from various solvents and mixed solvent systems,
24 5 5 chromatographic techniques, and counter current distribution, all of which are usually employed for this purpose.
The novel ani ihact'-ri i 1 agents of tho present invention are organic bases and thus are capable of forming acid-addition salts with a variety of organic and inorganic salt-forming reagents. Thus, acid-addition salts, formed by admixture of the antibacterial free base with up to three equivalents of an acid, suitably in a neutral solvent, are formed with such acids as sulfuric, phosphoric, hydrochloric, hydrobromic, sulfamic, citric, maleic, fumaric, tartaric, acetic, benzoic, gluconic, ascorbic, and related acids. The acid-addition salts of the antibacterial agents of the present invention are, in general, crystalline solids relatively soluble in water, methanol and ethanol but are relatively insoluble in non-polar organic solvents such as diethyl ether, benzene, toluene, and the like. For purposes of this invention, the antibacterial free bases are equivalent to their non-toxic acid-addition salts.
The antibiotics designated BM123fi^, BM123B2' BM123/i and BM123 are formed during the cultivation under controlled conditions of a new strain of an undetermined species of Nocardia. This new antibiotic producing strain was isolated from a garden soil, sample collected at Oceola, Iowa, and is maintained in the culture collection of the Lederle Laboratories Division, American Cyanamid Company, Pearl River, New York, as Culture No. BM123. A viable culture of the new microorganism has been deposited with the Culture Collection
Laboratory, Northern Utilization Research and Development Division, United States Department of Agriculture, Peoria, Illinois, and has been added to its permanent collection. It is freely available to the public in this depository under its accession No. NRRL 5646. Here in BM123P refers to a mixture in any proportion of ΒΜ123β·^ and ΒΜ123β2, and BM123/ refers to a mixture in any proportion of BM123'/'1 and BM123'/’2·
The following is a general description of the 10 microorganism Nocardia sp., NRRL 5646, based on diagnostic characteristics observed. Observations were made of the cultural, physiological, and morphological features of the organism in accordance with the methods detailed by Shirling and Gottlieb, Internat. Journ. of
Syst. Bacteriol, 1(5:213-240 (1966). The chemical composition of the culture was determined by the procedures given by Lechevalier et al., Advan. Appl. Microbiol. 14:· 47-72 (1971). The underscored descriptive colors and color chip designations are taken from Jacobson et al.,
Color Harmony Manual, 3rd ed. (1948), Container Corp, of America, Chicago, Illinois. Descriptive details are recorded in Tables I through V below.
Amount of Growth
Moderate on yeast extract, asparagine dextrose,
Benedict's, Bennett's, potato dextrose and
Weinstein's agars; light on Hickey and Tresner's, tomato paste, oatmeal, and pablum agars, and a trace of growth on inorganic salts-starch,
Kuster's oatflake, Czapek's solution, and rice agars.
2 4 5 5
Aerial Mycelium
Aerial mycelium whitish when present; produced only on yeast extract, asparagine dextrose, Benedict's, Bennett's, and potato dextrose agars.
Soluble Pigments
No soluble pigments produced.
Reverse Color
Colorless to yellowish shades.
Miscellaneous Physiological Reactions
No liquefaction of gelatin; nitrates reduced to nitrates in 7 days; melanoid pigments not formed on peptone-iron agar; no peptonization or curd formation in purple milk; NaCl tolerance in yeast extract agar >4% but <^7%; optimal growth temperature 32°C. Carbon source utilization, according to the Pridham and Gottlieb method [J. Bacteriol. 56:107-114 (1948)] as follows: Good utilization of glycerol, salicin, d-trehalose and dextrose; fair utilization of ί-inositol; and poor to non-utilization of d-fructose, maltose, adonitol, 1-arabinose, lactose, d-mannitol, d-melibiose, d-raffinose, 1-rhamnose, succrose and d-xylose.
Chemical Composition
The organism belongs to cell wall type IV, i.e. contains meso-2,6-diaminopimelic acid and has a Type A whole-cell sugar pattern, i.e., contains arabinose and galactose. Methylated whole cell extracts, when subjected to gas
I
4245 5 chromatography, showed fatty acid patterns similar to those produced by Nocardia asteroides ATCC 3308.
Micromorphology
Aerial mycelium arises from substrate mycelium as sparingly branched moderately long flexous elements that commonly terminate in elongated primitive spirals. The flexous elements are irregularly segmented into short elliptical to
' cylindrical sections (spores?) which disarticulate readily. The spiral terminal portions are less conspicuously segmented. Segments generally range 0.8-1.7 yvn x 0.3-0.5 pm, averaging 0.4 pm x 1.2 pm.
Diagnosis
The morphological characteristics of Culture No. BM123 are difficult to observe and interpret because of the poor development Of aerial mycelium on most media. Hence, considerable importance is attached, out of necessity, to the chemical analysis in determining the generic relationship of the organism. On the basis of the system proposed by Lechevalier et al., Culture No. BM123 contains meso-2,625 -diamihOpimelic aoid in its whole cells and sugar analysis shows arabinose and galactose to be present. Therefore, the culture belongs to cell wall type IV. A comparison of the gas chromatography pattern of Culture No. BM123 with that of Nocardia asteroides ATCC 3308
3 4 5 5 showed the two to be remarkably similar. Other characteristics of Culture No. BM123 that are in keeping with the Nocardia concept, are its fragmenting aerial growth on some media and the total absence of aerial growth on most media.
In view of the lack of adequate criteria for the characterization of Nocardia to the species level, no attempt has been made to make this determination. Therefore, Culture No. BM123 will be considered an undetermined species of
Nocardia until such a diagnosis is feasible.
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NRRL 5546
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- 10 424S5
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- 11 43455
TABLE II
Micromorphology of Nocardia sp. NRRL 5646
Medium Aerial Mycelium and/or Sporiferous Structures Yeast Extract Agar Aerial mycelium arises from substrate mycelium as sparingly branced, flexous elements that commonly terminate in elongated primitive spirals. The flexuous elements are irregularly segmented into short sections (spores?) which disarticulate readily. The spiral terminal portions are less conspicuously segmented. Segments generally range 0.8-1^7 Jim x 0.3-0.5 jim, averaging 0.4 jam x 1.2 jim.
- 12 42455
Iscellaneous Physiological Reaction of Nocardia sp. NRRL 5646
. '4 3 4 5 5
TABLE IV
Carbon Source Utilization Pattern of Nocardia sp. NRRL 5646 Incubation: 10 days Temperature: 32°C.
Carbon Source Utilization* Adonitol 0 1-Arabinose 0 Glycerol 3 d-Fructose 1 i-Inositol 2 Lactose 0 d-Manrtitol 0 Salicin 2 d-Melibiose 0 d-Raffinose 0 Rhamnose 0 Maltose 1 Sucrose 0 d-Trehalose 3 d-Xylose 0 Dextrose 3 Negative Control 0
*3-Good Utilization 1-Poor Utilization
2-Fair Utilization 0-No Utilization
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TABLE V
Chemical Composition of Nocardia sp. NRRL 564G
Cell Wall Type Major Constituents Type IV nieso-DAP, a rab inose, galactose
The production of BM1230 and BM123-/ is not limited to this particular organism or to organisms fully answering the above growth and microscopic characteristics which are given for illustrative purposes only.
In fact, mutants produced from this organism by various means such as exposure to X-radiation, ultra-violet radiation, nitrogen mustard, actinophages, and the like, may also be used. A viable culture of a typical such mutant strain has been deposited with the Culture Collection Laboratory, Northern Utilization Research and Development Division, United States Department of Agriculture, Peoria, Illinois, and has been added to its permanent collection under its accession number NRRL 8050. Although the cultural, physiological, and morphological features of NRRL 8050 are substantially the same as those of NRRL 5646, it produces enhanced amounts of BM123/” during aerobic fermentation. Also, NRRL 8050 varies from the parent NRRL 5646 as follows:
(a) slower reduction of nitrates to nitrites; and (b) production of a rosewood tan mycelial pigment on Bennett's and yeast extract agars.
The novel antibacterial agents of the present invention are, in general, crystalline solids of relatively limited solubility in non-polar solvents such as diethyl ether and hexane, but considerably more soluble in solvents such as water and lower alkanols. Anti5 biotics 1315123/-^ and BM123V<2 are structural isomers and may be represented by the following structural formulae:
- 16 42455
- 18 42455
The reductive alkylation of BM123’/', and ΒΜΙΣΙ/^ or BM123/2 with ketones takes place on the spermadine side-chain to form derivatives of the formula:
R
trans j| /¾
CH=CH-C-NH- (CII2) j-NH- (CIl,) .-NH-CH ^R3 wherein R is a moiety of the formulae:
and R2 and R-j are as hereinabove defined. The reductive alkylation of BM123/ζ BM123VZ1 or BM123/2 with aldehydes takes place on the spermadine side-chain to form mono-, di-, and tri-substituted derivatives of the formulae:
trans ||
R-// y-CH=CH-C-NII-(CH2)3~NH-(cfI2) 4_NHCH2_R1 0 /CBj-R].
/77\ trans || / R \ /CH=CH“C“NH- (ch2} 3-NH_ (ch2> 4~nx ch2-ri /7~~λ trans || _ y-CH=CH-C-NH-(CH2)3-N-(CHz)4-NH-CH2-R1 ch2-r1 trans || /CH2_Rl j^J^-CH^CH-C-NH- (CH2) 3-Nch2-Ri wherein R and R^ are as hereinabove defined.
The usefulness of the alkylated derivatives of BM123/Z is demonstrated by their ability to control systemic lethal infections in mice. Those new sub20 stances show high in vivo antibacterial activity in mice against Escherichia coli US311 when administered by a single subcutaneous dose to groups of Carworth Farms CF-1 mice, weight about 20 gm., infected intraperitoneally with a lethal dose of this bacteria in a IO-** trypticase soy broth TSP dilution Of a 5 hour TSP blood culture.
In Table VI below is set forth the in vivo activity of typical products of this invention (prepared from the indicated carbonyl reagents) against Escherichia coli US311 in mice. The activity is expressed in terms of the
ED50 or the dose in mg./kg. of body weight required to protect 50% of the mice against E. coli.
424337
ED50 in mg./kg.
Carbonyl· Reagent Employed Derivative Name of body weight
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'42455
by 1 tridecyl-BMl 2 3/
43455
42453
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Fermentation Process Selected to Produce Primarily ΒΜ123β and BM123y<.
Cultivation of Nocardia sp. NRRL 8050 may be carried out in a wide variety of liquid culture media. Media which are useful for the production of the antibiotics include an assimilable source of carbon such as starch, sugar, molasses, glycerol, etc.; an assimilable source of nitrogen such as protein, protein hydrolyzate, polypeptides, amino acids, corn steep liquor, etc.; and inorganic anions and cations, such as potassium, magnesium, calcium, ammonium, sulfate, carbonate, phosphate, chloride, etc. Trace elements such as boron, molybdenum, copper, etc.; are supplied as impurities of other constituents of the media. Aeration in tanks and bottles is provided by forcing sterile air through or onto the surface of the fermenting medium. Further agitation in tanks is provided by a mechanical impeller. An antifoaming agent, such as HodagCi FD82 may be added as needed.
Inoculum Preparation for ΒΜ123β and BM123/
Primary shaker flask inoculum of Nocardia sp.
NRRL 8050 is prepared by inoculating 100 milliliters of sterile liquid medium in 500 milliliter flasks with scrapings or washings of spores from an agar slant of the culture. The following medium is ordinarily used:
Bacto-tryptone Yeast extract Beef extract Glucose Water to gm. 5 gm. 3 gm.
gm. 1000 ml.
The flasks were incubated at a temperature from 25-29°C., preferably 28°C. and agitated vigorously
4245 5 on a rotary shaker for 30 to 48 hours. The inocula are then transferred into sterile screw cap culture tubes and stored at below 0°F. This bank of vegetative inoculum is used instead of slant scrapings for inocu5 lation of additional shaker flasks in preparation of this first stage of inoculum.
These first stage flask inocula are used to seed 12 liter batches of the same medium in 20 liter glass fermentors. The inoculum mash is aerated with sterile air while growth is continued for 30 to 48 hours.
The 12 liter batches of second stage inocula are used to seed tank fermentors containing 300 liters of the following sterile liquid medium to produce the third and final stage of inoculum:
Meat solubles 15 gm. Ammonium sulfate 3 gm. Potassium phosphate, dibasic 3 gm. Calcium carbonate 1 gnu Magnesium sulfate heptahydrate 1.5 gm. Glucose 10 gm. Water to 1000 ml.
The glucose is sterilized separately.
The third stage inoculum is aerated at 0.4 to 0.8 liters of sterile air per liter of broth per minute, and the fermenting mixture is agitated by an impeller driveh at 150-300 revolutions per minute. The temperat25 ure is maintained at 25-29°C., usually 28°C. The growth is continued for 48 to 72 hours, at which time the inoculum is used to seed a 3000 liter tank fermentation. Tank Fermentation for ΒΜ123β and liMI23~/
For the production of ΒΜ123β and BM12 3V/' in tank fermentors, the following fermentation medium is preferably used:
Meat solubles 30 gm. Ammonium sulfate u gm. Potassium phosphate, dibasic 6 gm. Calcium carbonate 2 gm. Magnesium sulfate heptahydrate 3 gm. Glucose 20 gm. Water to 1000 ml.
The glucose is sterilized separately.
Each tank is inoculated with 5 to 10% of third stage inoculum made as described under inoculum preparation. The fermenting mash is maintained at a temperature of 25-28°C., usually 26°C. The mash is aerated with sterile air at a rate of 0.3-0.5 liter of sterile air per liter of mash per minute and agitated by an impeller driven at 70 to 100 revolutions per minute. The fermentation is allowed to continue from 65-90 hours and the mash is harvested.
The invention will be described in greater detail in conjunction with the following specific examples.
Example 1
Inoculum Preparation for BM12 3(i and BM12 3
A typical medium used to grow the first and second stages of inoculum was prepared according to the following formula:
Bacto-tryptone Yeast extract Beef extract Glucose Water to gm. 5 gm. 3 gm.
gm. 1000 ml.
Two 500 milliliter flasks each containing 100 milliliters of the above sterile medium were inoculated with 5 milliliters each of a frozen vegetative inoculum from Nocardia sp. NRRL 8050. The flasks were placed on a rotary shaker and agitated vigorously for 48 hours at 28°C. The resulting flask inoculum was transferred to a 5 gallon glass fermentor containing 12 liters of the above sterile medium. The mash was aerated with sterile air while growth Was carried out for about 48 hours, after which the contents were used to seed a 100 gallon tank fermentor containing 300 liters of the following sterile liquid medium:
Meat solubles 15 gm. Ammonium sulfate 3 gm. Potassium phosphate, dibasic 3 gm. Calcium carbonate 1 gm. Magnesium sulfate heptahydrate 1.5 gm. Glucose 10 gm. Water to 1000 ml. The glucose is sterilized separately.
The third stage of inoculum mash was aerated 15 with sterile air sparged into the fermentor at 0.4 liters of air per liter of mash per minute. Agitation was supplied by a driven impeller at 240 revolutions per minute. The mash was maintained at 2 8 °C. and Hodag® FD82 was used as a defoaming agent. After 48 hours of growing time the inoculum mash was used to seed a 3000 liter fermentation.
Example 2
Fermentation Employing Nocardia sp. NRRL 8050 and Medium Favoring the Production of BM1238 and BM123yz.
A fermentation medium was prepared according to the following formula:
Meat solubles 30 gm. Ammonium sulfate 6 gm. Potassium phosphate, dibasic 6 gm. Calcium carbonate 2 gm. Magnesium sulfate heptahydrate 3 gm. Glucose 20 gm. Water to 1000 ml. The glucose is sterilized separately.
43455
The ternien.it ion medium waa sterilized .it l-hF'C.
with steam at 20 pounds pressure for 60 minutes. The pll of the medium after sterilization was 6.9. Three thousand liters of sterile medium in a 4000 liter tank fer5 mentor was inoculated with 300 liters of inoculum such as described in Example 1, and the fermentation was carried out at 26°C. using Hodag® FD82 as a defoaming agent. Aeration was supplied at the rate of 0.35 liter of sterile air per liter of mash per minute. The mash was agi10 tated by an impeller driven at 70-72 revolutions per minute. At the end of 67 hours of fermentation time the mash was harvested.
Example 3 isolation of BM123P and BM123y^
A 3000 liter portion of fermentation mash prepared as described in Example 2, pH 4.3, was adjusted to pH 7.0 with sodium hydroxide and filtered using 5-ύ diatomaceous earth as a filter aid. The cake was washed with about 1.00 liters of water anti discarded. The com20 bined fill rate anil wash was pumped upward through liiree parallel 8 1/4 x 48 stainless steel columns ..·<ι-·Ιι containing 15 liters of CM Sephadex'/ C-25 [Na4 ] resin (a cross-linked dextra-epichlox-ohydrin cation exchange gel available from Pharmacia Fine Chemicals, Inc.). The charged columns were washed with a total of about 390 liters of water and then developed with 200 liters of
17. aqueous sodium chloride followed by 560 liters of 5% aqueous sodium chloride. The 5% aqueous sodium chloride eluate was clarified by filtration through diatomaceous earth and the clarified filtrate passed through a 9 x
4.24 5 5
60 glass column containing 25 liters of granular
Darco® G-60 (20-40 mesh) (a granular activated carbon available from Atlas Chemical Industries, Inc.). The charged column was washed with 120 liters of water and then developed with 120 liters of 15% aqueous methanol followed by 340 liters of 50% aqueous methanol and then 120 liters of 50% aqueous acetone. The 15% aqueous methanol eluate was concentrated in vacuo to about 7 liters of an aqueous phase and the pH adjusted from 4.5 to 6.0 with Amberlite® IR-45 (OH-) resin (a weakly basic polystyrene-polyamine type anion exchange resin)> The resin was removed by filtration and the filtrate was concentrated in vacuo to about 1 liter and then lyophilized to give 38 grams of material consisting primarily of BM123U along with a small amount of BM123/” (primarily BM123V^2). The 50% aqueous methanol eluate was adjusted from pH 4.65 to 6.0 with Amberlite® IR-45 (OH-) resin. The resin was removed by filtration and the filtrate was concentrated in vacuo to about 6.3 liters and then lyophylized to give 213 grams of material consisting primarily of 33^123/^. The 50% aqueous acetone eluate was adjusted from pH 4.0 to 6.0 with Amberlite®
IR-45 (OH-) resin. The resin was removed by filtration and the filtrate was concentrated in vacuo to about 1.5 liters and then lyophylized to give 56 grams of impure BM123/'.
Example 4
Further Purification of BM123'/Z A slurry of CM Sephadex® C-25 (NH^] in 2% aqueous ammonium chloride was poured into a 2.6 centi36 meter diameter glass column to a resin height of approximately 62 centimeters. The excess 2% aqueous ammonium chloride was drained away and a 5.0 gram sample of BM123/* prepared as described in Example 3 was dissolved in about 10 milliliters of 2% aqueous ammonium chloride and applied to the column. The column was then eluted with a gradient between 6 liters each of 2% and 4% aqueous ammonium chloride. Fractions of about 75 milliliters each were collected automatically every 15 minutes. Antibiotic BM123/ was located by monitoring the column effluent in the ultra-violet and by bioautography of dipped paper disks on large agar plates seeded with Klebsiella pneumoniae strain AD. The majority of BM123V^ was located between fractions 71-107 inclusive.
One hundred thirty milliliters of granular Darco® G-60 (2040 mesh) was suspended in water, transferred to a glass column, allowed to settle and the excess water was allowed to drain away. Fractions 84-96 inclusive from the above CM Sephadex chromatography were combined and passed through the granular carbon column.
The charged column was washed with 600 milliliters of water and then developed with 1 liter of 50% aqueous acetone. The eluates, both of which contained BM123)/, were concentrated to aqueous phases in vacuo and lyophilized to give a total of 886 milligrams of BM123l/ as the hydrochloride salt. A microanalytical sample was obtained by subjecting the above material to a repeat of the above process.
Antibiotic BM123/X does not possess a definite melting point, but gradual decomposition starts in the vicinity of 200°C. Microanalysis of a sample equilibrated for 24 hours in a>72°F. atmosphere containing 23% relative humidity gave C, 39.44%; H, 6.10%; N, 16.19% Cl(ionic), 11.54%; loss on drying, 8.19%. In water
BM123'/' gave a U.V. absorption maximum at 286 nm with eJ·* = 250. The position of this maximum did not change lcm with pH. BM123Y^ had a specific rotation of [α]β = +71° (C - 0.97 in water).
Antibiotic BM123/^ exhibited characteristic 10 absorption in the infrared region of the spectrum at the following wavelengths: 770, 830, 870, 930, 980, 1035, 1105, 1175, 1225, 1300, 1340, 1370, 1460, 1510, 1555, 1605, 1660, 1740, 2950 and 3350 crrT^. A standard infrared absorption spectrum of BM123’/< prepared in a KBr pellet is shown in Figure 1 of the accompanying drawings. Example 5
Isolation of ΒΜ123)Λ
A slurry of CM Sephadex® C-25 [Na+] in 2% aqueous sodium chloride was poured into a 2.6 centimeter diameter glass column to a resin height of approximately 70 centimeters. The excess 2% aqueous sodium chloride was drained away and 4.11 gram of a sample containing primarily BM123/^ along with some BM123/^ and other impurities, prepared as described in Example 3, was dis25 solved in about 10 milliliters of 2% aqueous sodium chloride and applied to the column. The column was then eluted with a gradient between 4 liters each of 2% and 4% aqueous sodium chloride. Fractions of about 75 milliliters each were collected automatically every 15 min30 utes. Antibiotic BM123/Z was located by monitoring the
2 4 S 5 column effluent In the ultraviolet and by bioautography of dipped paper disks on large agar plates seeded with Klebsiella pneumoniae strain AD. The majority of 13(1123/^ was located between fractions 64-90 inclusive; the initial fractions (64-80) contained a mixture of ΒΜ123'/’ι and BM123vC, whereas the later fractions (81-90) contained essentially pure B!112 3V^.
One hundr ed milliliters of granular narco1'
G-60 (20-40 mesh) w.ih suspended in water , Lrunslerred lo a glass column, allowed to settle and the excess water was allowed to drain away. Fractions 81-90 inclusive from the above CM Sephadex chromatography were combined and passed through the granular carbon column. The charged column was washed with 500 milliliters of water and then developed with 500 milliliters of 10% aqueous methanol followed by 1 liter of 50% aqueous methanol.
The 50% aqueous methanol eluate, which contained the majority of ΒΜ123/^, was adjusted from pH 5.9 to 6.0 with Amberlite® IR-45 (Oil-1) resin. The resin was removed by filtration and the filtrate was concentrated in vacuo to an aqueous phase and lyophilized to give 294 milligrams of white amorphous BM123as the hydrochloride salt.
Antibiotic BM123does not possess a definite melting point, but gradual decomposition starts in the vicinity of 200°C. Microanalysis of a sample equilibrated for 24 hours in a 70°F. atmosphere containing 60% relative humidity gave C, 37.84%; 11, 5.73%; N,
.58%; Cl(ionic), 10.01%; loss on drying 10.45%. In methanol BM123Y^ gave a U.V. absorption maximum at 286 .43455 nm with E^m = 225. The position of this maximum did not change with pH. BM123had a specific rotation of +55° (C=0.803 in water).
Antibiotic BM123 exhibited characteristic absorption in the infrared region of the spectrum at the following wavelengths: 770, 830, 870, 930, 980, 1045, 1080, 1110, 1125, 1175, 1225, 1305, 1345, 1380, 1465, 1515, 1560, 1605, 1660, 1730, 2950 and 3350 cm-1.
A standard infrared absorption spectrum of Bt-1123’/^ pre10 pared in a KBr pellet is shown in Figure 2 of the accompanying drawings. A standard proton magnetic resonance spectrum of BM123/^ determined on a D20 solution in a 100 megacycle spectrometer is shown in Figure 4 of the accompanying drawings.
Example 6
Isolation of BM123Y^
A 25 gram sample containing primarily BM1232 and ΒΜ123β, prepared as described in Example 3, was dissolved in about 120 milliliters of 2% aqueous sodium chloride and applied to a column containing 1800 ml. of CM Sephadex® C-25 [Na+] in 2% aqueous sodium chloride.
The column was then eluted with a gradient between 20 liters each of 2% and 4% aqueous sodium chloride. The initial 12 liters of eluate was collected in a large bottle and discarded. Thereafter fractions of about 800 milliliters each were collected automatically every 40 minutes. Antibiotic BM123V” was located by monitoring the column fractions in the ultraviolet. The majority of BM123Y’/ was located between fractions 7-18 inclusive;
the initial fractions (7-15) contained essentially pure
2 4 5 5
DM123ancJ fche later fractions (16-18) contained a mixture of BM123)/^ and BM123
Six hundred milliliters of granular Darco®
G-60 (20-40 mesh) was suspended in water, transferred to a glass column, allowed to settle and the excess water was allowed to drain away. Fractions 7-15 inclusive from the above CM Sephadex chromatography were combined and passed through the granular carbon column. The charged column was washed with 3 liters of water and then developed with 3 liters of 10% aqueous methanol followed by 6 liters of 50% aqueous methanol. The 10% aqueous methanol eluate was adjusted from pH 5.8 to 6.0 with Amberlite® IH 45 (OH-) resin. The resin was removed by filtration and the filtrate was concentrated in vacuo to an aqueous phase and lyophilized to give 595 milligrams of white amorphous BM123Y/2 as the hydrochloride salt. The 50% aqueous methanol eluate was adjusted from pH 4.6 to 6.1 with Amberlite® IR 45 (OH-) resin.
The resin was removed by filtration and the filtrate was concentrated in vacuo to an aqueous phase and lyophilized to give 3.645 grams of slightly less pure white amorphous ΒΜ123/”2 as the hydrochloride salt.
Antibiotic BM123VZ2 does not possess a definite melting point, but gradual decomposition starts in the vicinity of 200eC. Microanalysis of a sample equilibrated for 24 hours in a 70°F. atmosphere containing 60% relative humidity gave C, 36.14%, Ii, 5.67%; N, IS.1%; Cl(ionic), 11.11%; loss on drying 10.87%. In methanol BM123Y^> gave a U.V. absorption maximum at 286 nm with = 220. The position of this maximum did not change
424 5 5 with pH. BM123y^ had a specific rotation of +60° (C=
0.851 in water).
Antibiotic BM123 exhibited characteristic absorption in the infrared region of the spectrum at the following wavelengths: 770, 830, 870, 950, 980, 1035, 1110, 1175, 1225, 1285, 1345, 1380, 1470, 1515, 1560, 1605, 1660, 1755, 2950 and 3350 cm-1. A standard infrared absorption spectrum of BM123 '/C, prepared in a KBr pellet is shown in Figure 3 of the accompanying drawings. A standard proton magnetic resonance spectrum of BM123 determined on a D2O solution in a 100 megacycle spectrometer is shown in Figure 5 of the accompanying drawings.
Example 7
Paper Partition and Thin Layer Chromatography of BM1238 and BM123/· The Bml23 antibiotics can be distinguished by paper chromatography. For this purpose Whatman No.
strips were spotted with a water or methanol solution of the substances and equilibrated for 1 to 2 hours in the presence of both upper and lower phases. The strips were developed overnight with the lower (organic) phase obtained from mixing 90% phenol:m-cresol:acetic acid:pyridine:water (100:25:4:4:75 by volume). The developed strips were removed from the chromatographic chamber, air dried for 1 to 2 hours, washed with ether to remove residual phenol and bioautographed on large agar plates seeded with Klebsiella pneumoniae strain AD. Representative Rf values are listed in Table VII below:
24 55
TABLE VII
Component RF
BM123'< 0.85
BM1230 0.50, 0.70
The β component was a mixture of two antibiotics using this system. BM1230 was composed of a major antibiotic (Rf = 0.50) called ΒΜ123β^ and a minor antibiotic (Rf = 0.70) called BM12302.
The BM123 antibiotics can also be distinguished by thin layer chromatography. For this purpose pre-coated Cellulose F® plates (0.10 millimeters thick), a form of thick layer cellulose supplied by EM Laboratories, Inc., Elmsford, New York, were spotted with a water solution of the substance to be chromatographed (about 20-40 micrograms per spot). The plates were developed overnight with the solvent obtained by mixing 1-butanol:water:pyridine:acetic acid (15:12:10:1 by volume). The developed plates were removed from the chromatographic chamber and air dried for about 1 hour. The antibiotics were detected by using either standard ninhydrin or Sakaguchi spray reagents. Representative Rf values are listed in Table VIII below:
TABLE VIIT
Component BM123/’ BM1230 Both BM123P components using this major component (Rf = minor component (Rf =
Rf
0.17, 0.23 0.08, 0.14 and BM123Y^ were a mixture of two system. BM1230 was composed of a 0.08) which was ΒΜ123β2 and a 0.14) which was ΒΜ123β2· The less polar component of BMlSs/* (Rf = 0.23) was BM123)/j_ and the more polar component (Rf = 0.17) was BMWlV^·
Example 8
General Procedure for Reductive Alkylation of Antibiotic _BM123<__
To a stirred solution of 100 mg. of antibiotic ΒΜ123)^ in 20 ml. of methanol is added 5 ml. (or 5 g.) of the appropriate aldehyde or ketone and 100 mg. of sod·ium cyanoborohydride. The pH of the resulting solution is maintained at about 7.0 with 0.1N methanolic hydrogen chloride over a 3 to 24 hour period. The reaction is monitored by thin layer chromatography to the disappearance of BM123l/\. The reaction mixture is then filtered and the filtrate is evaporated to dryness. The residue is triturated with 3 ml. of methanol and filtered. The filtrate is diluted with 50 ml. of acetone and the precipitate which forms is removed by filtration and dried. The methanol solvent may be replaced by 20 ml· of water wherever the starting aldehyde or ketone is water soluble.
Example 9
Preparation of methy1-BM123Z'
To a solution of 1.0 g. of BM123Y/” and 2.5 ml.
of a 37% aqueous formaldehyde solution in 50 ml. of water is added, protionwise, 400 mg. of sodium cyanoborohydride The pH of the reaction mixture was maintained at 7.0 with IN hydrochloric acid during this addition. The reaction mixture was stirred an additional ten minutes at room temperature and then evaporated to dryness in vacuo.
The residue was triturated with 20 ml. of methanol, filt44 ered and the filtrate diluted with 250 ml. of acetone. The product which precipitated was removed by filtration and dried; yield, 667 mg.
Example 10
Preparation of isopropyl-BM12 37^
To a solution of 200 mg. Of BM1237Z in 30 ml.
of methanol was added 5 ml. of acetone. To this solution was added 139 mg. of sodium cyanoborohydride and the reaction mixture was stirred at room temperature for 30 minutes. During this time the pH of the reaction mixture was maintained between 7.4 and 7.8 by the addition of 0.1N methanolic hydrogen chloride. The small amount of precipitate which had dormed was removed by filtration and the filtrate was evaporated to dryness in vacuo. The residue was triturated with two ml. of methanol and filtered. The filtrate was diluted with 100 ml. of acetone and the solid product that separated was removed by filtration and dried; yield, 184 mg.
Example 11
Preparation of 8-phenylethyl-BM123Z
To a solution of 200 mg. of BM123/* in 15 ml.
of water and 25 ml. of acetonitrile was added a solution of 2 ml. of phenylacetaldehyde in 4 ml. of ethanol. To this was added 103 mg. of sodium cyanoborohydride.
The reaction mixture was stirred at room temperature for thirty minutes during which time the pH of the mixture was maintained at 7 with 0.2N hydrochloric acid. The reaction mixture was then filtered and the filtrate was evaporated to dryness in vacuo. The residue was triturated with two ml. of methanol and filtered. The fil45 trate was diluted with 100 ml. of acetone and the product that separated was removed by filtration and dried; yield, 180 mg.
Example 12
Preparation of l,3,3-trimethylbutyl-BM123/
To a solution of 200 mg. of BM123/ hydrochloride in 50 ml. of methanol was added 3 ml. of 4,4-dimethyl-2-pentanone and 106 mg. of sodium cyanoborohydride. The reaction solution was maintained at pH 7 by the dropwise addition of methanolic hydrogen chloride. The reaction was stirred at room temperature for 18 hours and filtered. The filtrate was evaporated to dryness in vacuo. The residue was dissolved in 3 ml. of methanol, diluted with 50 ml. of acetone and filtered, yield 125mg.
Example 13
Preparation of l-methylphenethyl-BM123/
To a solution of 200 mg. of BM1231/ in 50 ml.
of methanol was added 5 ml. of phenylacetone. To this solution was added 170 mg. of sodium cyanoborohydride and the reaction mixture stirred at room temperature for 3 and a half hours. During this time the pH of the reaction mixture was maintained at 7.0 with methanol saturated with hydrogen chloride gas. Reaction mixture was concentrated to about 5 ml. volume, diluted with two ml. of methanol, and filtered. Filtrate was poured into 100 ml. of acetone and the solid product that separated was removed by filtration and dried; yield, 233 mg.
Example 14
Preparation of l-methylnonyl-BM123’/'
Sodium cyanoborohydride (100 mg.) was added to a solution of BM123Y^ (200 mg.) and 2-decanone (I ml.) in 40 ml. of methanol. The pH of the solution was adjusted to 7.0 and maintained at 7.0 I 0.2 by tiie addition of 0.1N methanolic hydrogen chloride as necessary. After 19.5 hours the reaction mixture was filtered and the filtrate was concentrated iri vacuo at 30’C. The residue was slurried in 5 ml. of methanol and filtered. Tiie filtrate was added to 50 ml. of acetone. The off-white solid which precipitated was collected by filtration, washed with acetone, and dried in vacuo. The yield of crude l-methylnonyl-BM123'/z” was 167 mg.
Example 15
Preparation of 1,3-dimethylbutyl-BM123yz'
To a solution of 210 mg. of BM123V^ in 50 ml. of methanol was added 5 ml . of methyl, isobutyl ketone.
To this solution was added 166 mg. of sodium eyanoborohydride and the reaction mixture stirred at room temperature for five hours. During this time the pH of the reaction mixture was maintained at 7.0 with methanol saturated with hydrogen chloride gas. Reaction mixture was evaporated to dryness in vacuo. The residue was triturated with two ml. of methanol and filtered. The filtrate was diluted with 100 ml. of acetone and the solid product that separated was removed by filtration and dried; yield, 210 mg.
Example 16
Preparation of 2-norbornyl-BM123/^
Sodium cyanoborohydride (100 mg.) was added to a solution of BM123Y^ (200 mg.) and 2-norbornanone
4345 5 (400 mg.) in 40 ml. of methanol. The pH of the solution was adjusted to 7.0 with 0.1N methanolic hydrogen chloride. The pH was maintained at 7,0 ί 0.2 by the addition of 0.1N hydrogen chloride as necessary. After
21.5 hours the reaction mixture was filtered and the filtrate was concentrated in vacuo at 35°C. The residue was slurried in*5 ml. of methanol and filtered. The filtrate was added to 50 ml. of acetone. The off-white solid which precipitated was collected by filtration, washed with acetone and dried in vacuo. The yield of crude 2-norbornyl-BM123/Z was 175 mg.
Example 17
Preparation of isopropyl-BM123'/^
A mixture of 50 mg. of BM123/Z^, 5 ml. of ace15 tone and 60 mg. of Sodium cyanoborohydride in 35 ml. of methanol was stirred at room temperature for 40 minutes. The pH of the solution was maintained at 7 by the dropwise addition of a methanolic hydrogen chloride solution. The mixture was evaporated to dryness in vacuo. The resi20 due was triturated with 5 ml. of methanol and the resulting solution was diluted with 50 mb. of acetone? yield, 49 mg.
Example 18
Preparation of isopropyl-BM123
A mixture of 41 mg. of BMl23y<2, 5 ml. of acetone and 50 mg. of sodium cyanoborohydride in 35 ml. of methanol Was stirred at room temperature for 40 minutes. The pH of the solution was maintained at 7 by the dropwise addition of a methanolic hydrogen chloride solution (saturated). The mixture was filtered and evaporated
2 4 5 5 to dryness in vacuo. The residue was triturated with 5 ml. of methanol and the resulting solution was diluted with 50 ml. of acetone; yield, 46 mg.
Example 19
Preparation of 1-methyl-2-phenyl-ethyl-BM123Z2
A mixture of 200 mg. of BMI23Z2, 5 ml. of phenyl-acetone and 170 mg. of sodium cyanoborohydride in 50 ml. of methanol was stirred at room temperature for 3 hours and 45 minutes. During this time the pll of the reaction mixture was maintained at 7 with dropwise addition of a methanolic hydrogen chloride solution (saturated). The mixture was evaporated to dryness in vacuo. The residue was triturated with 5 ml. of methanol and the resulting methanol solution was diluted with approximately 50 ml. of acetone; yield 233 mg.
Example 20
Preparation of (2-ethylcyclopentyl) BM123)/
A solution of 200 mg. of BM123)Z, 3 ml. of 2-ethylcyclopentanone and 101 mg. of sodium cyanoborohydride in 50 ml. of methyl alcohol was stored at room temperature for 18 hours. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol.
The reaction mixture was evaporated to dryness. The residue was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield, 157 mg.
Example 21
Preparation of 3,5-diinethylcyclohexyl BM123/
A solution of 200 mg. of BM123Z, 5 ml. of
24 5 5
3,5-dimethylcyclohexanone and 200 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 1 hour. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol.
The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield 200 mg.
Example 22
Preparation of 2,4-dimethylcyclopentyl BM123/'
A solution of 206 mg. of Bf-1123/^, 3 ml. of
2,4-dimethyloyclopentanone and 104 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 6 hours. During this time the pH of
IS the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol. The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield 101 mg.
. Example 23
Preparation of 2-ethylcyclohexyl BM123Y^
A solution of 200 mg. of BM123/S 5 ml. of 2-ethylcyclohexanone and 213 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temp25 erature for 3 hours. During this time the pH of the solution was maintained at 7 with the addition of saturated hydrogen chloride in methanol. The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield
200 mg.
- 50 <13 4 K S
Example 24
Preparation of 3-methylcyclohexyl BM123>/
A solution of 200 mg. of BM123/, 1.5 ml. of
3-methyIcyclohexanone and 200 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 2 hours. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol. The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield 200 mg.
Example 25
Preparation of 2,4,4-trimethylcyclopentyl BM123/'
A solution of 200 mg. of ΒΜΐ23/\ 5m]. of
2,4,4-trimethylcyclopentanone and 179 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 24 hours. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol. The reaction was triturated with 3 ml. of methanol, filt ered and the filtrate was diluted with 40 ml, of acetone, yield 176 mg.
Example 26
Preparation of 2-propylcyclohexyl BM123VZ
A solution of 200 mg. of BM123/, 3 ml. of 2-propylcyclohexanone and 157 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 4 hours. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol.
4 2 4 5 5
The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield 75 mg.
Example 27 5 Preparation of 2-methylcyclopentyl BM123>^
A solution of 211 mg. of BM123/<, 3 ml. of
2-methylcyclopentanone and 98 mg. of sodium cyanoborohydride in 50 ml. of methanol was stored at room temperature for 3.5 hours. During this time the pH of the solution was maintained at 7 with the addition of a saturated solution of hydrogen chloride in methanol. The reaction was triturated with 3 ml. of methanol, filtered and the filtrate was diluted with 40 ml. of acetone, yield 157 mg.
Claims (4)
1. A compound select ed from the group consisting of those of the formulae: trans η R-// \\_ cii=CH-C-NH-(CIIj) 3 -HII-(Clip,. -Nil GSIa-li, (I) t) .CIb-Ri /T7\ trans I, R-C CH=CH-C-NH’(CHj 3 -Nfl-(C!h)i.-N \ CHa-Rt (XI) 0 CHn-R, ΖΓλ trans „ | 1 R-\ _ >CH 3 -N-(Cll..),rNH-CH 2 -Ri (Iii) 0 CIU-R, rn r χρ-Λ trans ι ^t.il 2 -Ki R-/ Z ')— CH=Cll“C-NH -(Clli) 3-N-(Cil 2 )z,-N W CIU-R, ( IV) therein R is a moiety selected Ing of those of the formulae: from the group consist- - 53 42455 lo wherein is selected from the group consisting of hydrogen, lower alkyl, halo substituted lower alkyl, lower alkenyl, phenyl, monosubstituted phenyl, phenyl lower alkyl, 2-furyl, methyl substituted 2-furyl, 2-thineyl, methyl substituted 2-thineyl, 2-pyrryl, methyl substituted 2-pyrryl, 2-pyridyl and 2-quinolyl; and the pharmacologically acceptable acid-addition salts thereof.
2. A compound selected from the group consisting of those Of the formula: trans || R, CH=CH-C-NH*· (CH 2 ) 3NH- (CH 2 ) 4 -NH-CH^ wherein R is a moiety selected from the group consisting of those of the formulae: nh 2 nh 2 (A) and - 54 4 2 15 5 (Β) wherein R 2 is selected from the group consisting of lower alkyl, halo substituted lower alkyl and phenyl 5 lower alkyl; R^ is selected from the group consisting of lower alkyl, halo substituted lower alkyl, lower alkenyl, lower eycloalkyl, phenyl, monosubstituted phenyl, phenyl lower alkyl and monosubstituted phenyl lower alkyl; and R 2 and taken together with the 10 associated methylidyne group is cyclopentyl, mono-lower alkyl substituted cyclopentyl, di-lower alkyl substituted cyclopentyl, tri-lower alkyl substituted cyclopentyl, cyclohexyl, mono-lower alkyl substituted cyclohexyl, di-lower alkyl substituted cyclohexyl or tri 15 -lower alkyl substituted cyclohexyl; and the pharmacologically acceptable acid-addition salts thereof.
3. The process of preparing compounds of the formulae: trans || - CII=CH-C-NH- (CHa) 3 -Nli-(CHa) u-NH-CHa-Ri 20 (T) II s -C-NII-(Cll,.) .,-NII-(CH ? ) »-11 Clb-lh (II) - 55 43455 ΖΓΑ tl ail!! II I >-GH=Gti-G-NII- (Clip) 3 -W- (Gll ; .) ,,-NU-CH z -R» (IXX) 0 CH Z -R X trans || | CHa-R x •CH=GH-C-NH- (CHa)3—N- (CII 2 ) »-Νζ CHa-Ri (XV) wherein R·^ is hyd rogen, lower alkyl, halo substituted lower alkyl, lower alkenyl, phenyl, monosubstituted phenyl, phenyl lower alkyl, 2-furyl, methyl substituted 2-furyl, 2-thineyl, methyl substituted 2-thienyl, 2-pytryl, methylsubstituted 2-pyrryl, 2-pyridyl or 2-quinolyland R is a moiety of the formulae: - 56 4 2 4 5 5 which comprises alkylating an amine of the formula: o r-κ trans || R Cn=Cn-C-NiI “ (Ct, 2 ) 3 -nh (CH 2 ) 4 ~NH 2 wherein R is as hereinabove defined with an aldehyde of the formula: Rj-CHO wherein R^ is as hereinabove do fined in the presence of a reducing agent in a solvent inert; to the reactants for a period of time sufficient for a substantial degree of reductive alkylation to take place.
4. The process of preparing compounds of the formula: ΖΓΛ trans || R-/ Z _y-CH=CH-C-NH- (CH 2 ) 3 -NH- (CH 2 ) 4 ~N~CH Ro wherein R 2 is lower alkyl, halo substituted lower alkyl or phenyl lower alkyl; R 3 is lower alkyl, halo substituted lower alkyl, lower alkenyl, lower cycloalkyl, phenyl, monosubstituted phenyl, phenyl lower alkyl or monosubstituted phenyl lower alkyl; and R 2 and R 3 taken together is tetramethylene, pentamethylene or hexamethylene; and R is a moiety of the formulae: - 57 4 2455 which comprises alkylating an amine of the formula: trans CH=CH-C-NH-(CH 2 ) 3 -NH-(CH 2 ) 4 -NH 2 wherein R is as hereinabove defined with a ketone of the formula: · II R 2 -c-R 3 wherein R 2 and R 3 are as hereinabove defined in the presence of a reducing agent in a solvent inert to the reactants for a period of time sufficient for a substantial degree of reductive alkylation to take place.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US52986274A | 1974-12-05 | 1974-12-05 | |
US61297575A | 1975-09-12 | 1975-09-12 |
Publications (2)
Publication Number | Publication Date |
---|---|
IE42455L IE42455L (en) | 1976-06-05 |
IE42455B1 true IE42455B1 (en) | 1980-08-13 |
Family
ID=27063126
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
IE2651/75A IE42455B1 (en) | 1974-12-05 | 1975-12-05 | Alkylated derivatives of antibiotic bm123 |
IE1836/76A IE44567B1 (en) | 1974-12-05 | 1975-12-05 | Antibiotic bm123 and production thereof |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
IE1836/76A IE44567B1 (en) | 1974-12-05 | 1975-12-05 | Antibiotic bm123 and production thereof |
Country Status (25)
Country | Link |
---|---|
JP (1) | JPS51125279A (en) |
AR (1) | AR215581A1 (en) |
BG (1) | BG26404A3 (en) |
CA (1) | CA1072547A (en) |
CH (1) | CH626895A5 (en) |
DD (1) | DD124602A5 (en) |
DE (1) | DE2553927C3 (en) |
DK (1) | DK547975A (en) |
EG (1) | EG12248A (en) |
FI (1) | FI753415A (en) |
FR (1) | FR2293212A1 (en) |
GB (1) | GB1536153A (en) |
GR (1) | GR58534B (en) |
HK (1) | HK80779A (en) |
HU (1) | HU177070B (en) |
IE (2) | IE42455B1 (en) |
IL (1) | IL48476A (en) |
LU (1) | LU73930A1 (en) |
NL (1) | NL165469C (en) |
NO (1) | NO754011L (en) |
NZ (1) | NZ179276A (en) |
RO (1) | RO74426B (en) |
SE (1) | SE7513707L (en) |
YU (1) | YU304975A (en) |
ZM (1) | ZM16475A1 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA1087537A (en) * | 1975-08-21 | 1980-10-14 | John H. E. J. Martin | Antibiotic bm123 and production thereof |
US4112219A (en) * | 1977-07-13 | 1978-09-05 | American Cyanamid Company | Acyl derivatives of antibiotic BM123γ |
-
1975
- 1975-11-14 IL IL48476A patent/IL48476A/en unknown
- 1975-11-18 NZ NZ179276A patent/NZ179276A/en unknown
- 1975-11-25 GR GR49451A patent/GR58534B/en unknown
- 1975-11-26 CA CA240,497A patent/CA1072547A/en not_active Expired
- 1975-11-27 ZM ZM164/75A patent/ZM16475A1/en unknown
- 1975-11-27 AR AR261375A patent/AR215581A1/en active
- 1975-11-27 NO NO754011A patent/NO754011L/no unknown
- 1975-12-01 DE DE2553927A patent/DE2553927C3/en not_active Expired
- 1975-12-03 RO RO84081A patent/RO74426B/en unknown
- 1975-12-03 YU YU03049/75A patent/YU304975A/en unknown
- 1975-12-03 LU LU73930A patent/LU73930A1/xx unknown
- 1975-12-04 DK DK547975A patent/DK547975A/en not_active Application Discontinuation
- 1975-12-04 CH CH1581275A patent/CH626895A5/en not_active IP Right Cessation
- 1975-12-04 FI FI753415A patent/FI753415A/fi not_active Application Discontinuation
- 1975-12-04 SE SE7513707A patent/SE7513707L/en not_active Application Discontinuation
- 1975-12-04 FR FR7537174A patent/FR2293212A1/en active Granted
- 1975-12-04 HU HU75AE456A patent/HU177070B/en unknown
- 1975-12-05 JP JP50144097A patent/JPS51125279A/en active Pending
- 1975-12-05 IE IE2651/75A patent/IE42455B1/en unknown
- 1975-12-05 IE IE1836/76A patent/IE44567B1/en unknown
- 1975-12-05 NL NL7514234.A patent/NL165469C/en not_active IP Right Cessation
- 1975-12-05 DD DD189912A patent/DD124602A5/xx unknown
- 1975-12-05 BG BG031708A patent/BG26404A3/en unknown
- 1975-12-05 GB GB50093/75A patent/GB1536153A/en not_active Expired
- 1975-12-06 EG EG75719A patent/EG12248A/en active
-
1979
- 1979-11-22 HK HK807/79A patent/HK80779A/en unknown
Also Published As
Publication number | Publication date |
---|---|
ZM16475A1 (en) | 1977-03-21 |
DE2553927A1 (en) | 1976-06-16 |
BG26404A3 (en) | 1979-03-15 |
AU8659675A (en) | 1977-06-02 |
CA1072547A (en) | 1980-02-26 |
GR58534B (en) | 1977-10-31 |
SE7513707L (en) | 1976-06-08 |
AR215581A1 (en) | 1979-10-31 |
YU304975A (en) | 1982-05-31 |
FR2293212B1 (en) | 1980-06-27 |
IL48476A0 (en) | 1976-01-30 |
DE2553927C3 (en) | 1980-07-03 |
FR2293212A1 (en) | 1976-07-02 |
NZ179276A (en) | 1978-06-20 |
RO74426A (en) | 1983-04-29 |
NL165469B (en) | 1980-11-17 |
NL7514234A (en) | 1976-06-09 |
JPS51125279A (en) | 1976-11-01 |
DD124602A5 (en) | 1977-03-02 |
DK547975A (en) | 1976-06-06 |
HU177070B (en) | 1981-07-28 |
NO754011L (en) | 1976-06-09 |
EG12248A (en) | 1978-12-31 |
IE42455L (en) | 1976-06-05 |
LU73930A1 (en) | 1976-07-01 |
CH626895A5 (en) | 1981-12-15 |
GB1536153A (en) | 1978-12-20 |
FI753415A (en) | 1976-06-06 |
RO74426B (en) | 1983-04-30 |
DE2553927B2 (en) | 1979-10-18 |
IE44567B1 (en) | 1982-01-13 |
HK80779A (en) | 1979-11-30 |
IL48476A (en) | 1980-12-31 |
NL165469C (en) | 1981-04-15 |
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