JPH0526799B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to new antitumor antibiotics and their production and recovery. The antitumor antibiotic of the present invention, BBM-1644, is a new member of the protein antitumor antibiotic family exemplified by neocarzinostatin, macromycin, and auromomycin. Neocardinostatin (also known as dinostatin) is an acidic protein polymer with a molecular weight of 10,700, consisting of a single polypeptide chain of 109 amino acids cross-linked by two disulfide bridges. be. Streptomyces cardinostaticus var. neocarzinostaticus
The production of neocarzinostatin by fermentation of strains of N. carzinostaticus var .
(1965). The amino acid sequence of neocarzinostatin can be found in Cancer Treatment
Reviews 6 :239-249 (1979). Macromomycin is a neutral or slightly acidic polypeptide with an approximate molecular weight of approximately 15,000. Streptomyces macromomyceticus (NIHJ
The production of macromycin by fermentation of Mac-8-42) is described in US Pat. No. 3,595,954 and J. Antibiotics
21:44-49 (1968). Purification of macromycin and characterization data for purified compounds can be found in J. Antibiotics 29:415-423 (1976). Auromomycin has a molecular weight of approximately 12,500 and a pH of
It is a weakly acidic polypeptide with an isoelectric point of 5.4.
Consists of 16 different amino acids. Streptomyces macromomyceticus
The isolation of auromomycin from the culture fluid of Macromomyceticus and the properties of the purified product are described in J. Macromomyceticus.
Antibiotics 32 :330-339 (1979). BBM-1644 can be distinguished from known polypeptide antitumor antibiotics such as neocarzinostatin, macromomycin, and auromomycin by molecular weight, amino acid content, and paper electrophoresis. According to the present invention, a new strain of Actinomaszula, named Actinomaszula sp. BBM-1644 is produced by culturing BBM-1644 until a substantial amount of BBM-1644 is produced and optionally recovering BBM-1644 from the culture medium.
A novel protein antitumor antibiotic designated 1644 is provided. The present invention provides BBM- as dilute solution, crude concentrate, crude solid and purified solid.
Includes 1644 antibiotics. Figure 1 shows the infrared absorption spectrum of BBM-1644 (KBr pellet). Figure 2 shows water, 0.01N HCl and 0.01N NaOH
The ultraviolet absorption spectrum of medium BBM-1644 is shown. The present invention relates to a novel protein anti-tumor antibiotic herein named BBM-1644 and its production by fermentation of a new strain of Actinomaszula named strain Actinomaszula sp. H710-49. The above fungus was isolated from soil samples collected in West Germany. A biologically pure culture of this bacterium is
Commissioned by The American Type Culture Collection in Washington, DC, ATCC
It was added to its permanent microbial collection as 39144. BBM-1644 inhibits the growth of various Gram-positive and acid-fast bacteria. This antibiotic also shows phage inducibility in lysogens and P388 in mice.
Blocks the spread of lymphoid and solid tumors such as leukemia. Therefore, this new antibiotic can be used as an antibacterial agent or an antitumor agent to suppress tumors in mammals. Microorganisms Actinomycete strain No. H710-49 was isolated from a soil sample and prepared as a biologically pure culture for characterization by conventional methods. H710−49 strain is
Forms both sublayer and aerial hyphae. The lower hyphae are long and branched and cannot be cut into short mycelia. Short spore chains are born at the tips or uniaxial branches of aerial hyphae. Spore chains have 2 to 15 spores in the chain (mostly 4 to 8 spores) and are straight, hook-shaped or ring-shaped in shape. The spore has a shaped surface;
Oval or oval shape (0.5-0.6 x 0.7-1.2μm)
and has a round or pointed end. Mature spores are often separated by hollow hyphae. Terminal swelling of hyphae is observed on lower hyphae in Czapek agar or Bennet agar. No mobile spores, sporangia or hardened granules are seen in any of the media examined. Unlike the common species of Streptomyces,
Strain H710-49 grows slowly and forms poor aerial mycelia in chemically defined media and natural organic media. The color of aerial mycelia is white, and oatmeal agar, inorganic salts - starch agar and glycerol -
After sporulation in asparagine agar, the color changes to pink. The color of the mass of lower air threads is colorless, yellow, reddish-brown, or dark gray-brown. Melanoid pigments are not produced, but
Glycerol-asparagine agar, tyrosine agar, and glycerol, L-arabinose, D
- Pridum supplemented with any one of xylose, L-rhamnose, D-glucose, D-fructose, trehalose and D-mannitol -
Lemon yellow diffusible pigment is seen in Gottlieb's basal agar. H710−49 is 20℃, 28℃ and 37
It grows at 10°C or 41°C. Sensitive to 10% NaCl, but 7
% but not 0001% is resistant to lysozyme. D-galactose, D-mannose, sucrose, raffinose and inositol are not utilized by this strain. The culture and physiological characteristics of strain H710-49 are shown in Tables 1 and 2, respectively.
The type of carbon source utilization by this strain is shown in Table 8. Table 1 Culture characteristics of strain H710-49 ^* Tyrosine-yeast extract broth (ISP No.1)
Poor or moderate; hairy, smeared, unpigmented sucrose-nitrate agar G slight (Tsapetsk agar)
R Colorless to pale yellow-orange (73) ^*** A Slight; white (263) D None glucose-asparagine G Poor agar R Yellow-white (92) A Poor; white (263) to pale yellow-orange (31) D Bright yellow (83) Inorganic salt - starch agar (ISP No.4)
G Poor to moderate R Colorless to deep yellow (85) A Poor; red-white (9) to pale yellow-red (31) D None Tyrosine Agar (ISP No. 7) G Moderate R Brown-orange (54) to moderate Reddish brown (43) A Poor; white (263) to pale yellow (89) D Strong yellow (84) Normal agar G Poor to moderate R Yellowish white (92) to medium tan (77) A Poor; white ( 263) D None Yeast Extract - Malt Extract Agar (ISP No. 2)
G Moderate R Dark yellow (88) or dark brown (59) A Slight; white (263) D Pale olive brown (94) Oatmeal agar (ISP No. 3) G Poor R Colorless A Poor; white (263) or red-white Color (9) D None Bennet agar G Medium R Gray yellowish brown (80) to dark grayish brown (62) A Very slight; white (263) D Medium olive brown (95) Peptone-yeast extract-iron agar (ISP No.
6) G Poor R Gray-yellow (90) to dark gray-brown (62) A Poor; white (263) D None to medium tan (77) * Observation after 3 weeks of incubation at 28°C ** Abbreviation: G- Growth; R- Back color; A- Aerial mycelia; D- Diffusible pigment *** Colors and numbers in the box are "Kelly, K.
L. and DB jacket: ISCC-NBS color
−name charts illustrated with Centroid
Colors U.S. Department of Commerce Circular 553, Washington, D.C.
DC, November 1975” [Table] No growth ST extract agar
Table 3 Utilization of carbon sources by strain H710-49 ^* Glycerol + D(-)-arabinose - L(+)-arabinose + D-xylose + D-ribose + L-rhamnose + D-glucose + D-galactose - D- Fructose + D-mannose - L(-)-sorbose - sucrose - lactose - cellobiose + melibiose - trehalose + raffinose - D(+)-melezitose - soluble starch + cellulose - dulcitol - inositol - D-mannitol + D-sorbitol - Salicin - * Observation after 3 weeks of incubation at 28°C. Basal medium: Priddam-Gotzlieb mineral medium The purified cell wall of strain H710-49 contains mesodiaminopimelic acid but does not contain glycine. Whole cell hydrolyzate was prepared using mesulose (3-O-methyl-D
-galactose), glucose, ribose and a small amount of mannose. The cell wall composition and total cell sugar components of strain H710-49 indicate that this strain has a cell wall
Indicates that it belongs to type _B. The above-mentioned characteristics of strain H710-49 are similar to those of the genus Actinomasula. J. Gen． Microbiol .
112:95-111 (1979) According to the numerical taxonomy of Actinomaszula and related actinomycetes by Nakagutsudfellow et al., most of the Actinomaszula species of soil origin are
It is classified as strain group No. 7 out of the 14 bacterial strain groups described. Strain No. H710-49 is most related to strains of strain group 7. Nonomura and Ohara are
J. Ferment ． Technol ． 49 :904-912 (1971)
We report five species of saprophytic fungi in the genus Actinomasula,
Nonomura [ J. Ferment ． Technol ． 52 :71-77
(1974)] and Preobrazhenskaya et al. [ Actnomycetes and Related Organisms]
12:30-38 (1977)] described the identification and classification of Actinomasula species. As a result of comparison with known Actinomazura species described in the literature, H710−49
Strains include A. roseola, A. salmonea, A. vinacea, or A. corallina as described in the cited reference by Preobrazhenskaya et al. above and in Japanese Publication No. 55/94391. )
It is thought to belong to a new species of Actinomazura that resembles the. For the production of BBM-1644, the present invention uses a specific bacterial strain Actinomasula sp. strain H710-49 (ATCC 39144).
Although described in detail, it is not limited to this microorganism or to the microorganisms completely described by the culture characteristics disclosed herein. The present invention relates to H710-49 strain and its natural and artificial
It is specifically intended to include all BBM-1644 producing variants and mutants. Antibiotic Production The BBM-1644 antibiotic of the present invention is produced by a BBM-1644 producing strain of the genus Actinomasula, preferably ATCC.
It can be produced by culturing a strain of Actinomaszula sp. having the identifying characteristics of 39144 or its mutants in a conventional aqueous medium. The fungus grows in a normal medium containing known actinomycete nutritional sources, ie assimilable carbon and nitrogen sources plus any inorganic salts and other known growth factors. Deep aerobic conditions are preferably used for the production of large quantities of antibiotics, although surface cultures and bottles can also be used for production of limited quantities. General procedures used for culturing other actinomycetes are applicable to the present invention. The normal medium contains glycerol, L(+)-arabinose, D-xylose, D-ribose, D-glyose, D-fructose, soluble starch, D
- Should contain a suitable assimilable carbon source such as mannitol or cellobiose. As a nitrogen source, ammonium chloride, ammonium sulfate, uric acid, ammonium nitrate, sodium nitrate, etc. may be used alone or peptone, meat extract, yeast extract, corn staple, etc.
It can be used in combination with organic nitrogen sources such as liquor, soybean flour, cottonseed floors, etc. If necessary, common inorganic salts may also be added to provide sources of sodium, potassium, calcium, ammonium, phosphates, sulfates, chlorides, bromides, carbonates, zinc, magnesium, manganese, cobalt, iron, etc. It's fine. Production of BBM-1644 antibiotics can be carried out at any temperature at which sufficient growth of the producing bacteria occurs, e.g.
It can be carried out at 20-37°C, conveniently at 27°C.
Performed at temperatures around ~32°C. Optimum production is usually obtained after an incubation period of about 6-7 days in shake flasks. When carrying out tank fermentations, it is usually desirable to obtain a broth-grown inoculum by inoculating the broth culture with bacterial slants or soil or lyophilized cultures. After obtaining the active inoculum in this way, it is transferred aseptically to a fermentation tank medium. Antibiotic production is performed by Bacillus subtilis M45
(Rec ^- mutant bacteria; Mutation Res . 16:165-174
(1972)] can be monitored by paper disc-agar diffusion quantification using as test bacteria. Isolation and Purification When the fermentation is complete, after separating the solid portion by filtration or centrifugation, BBM-1644 is primarily present in the liquid portion of the fermentation broth. Thus, the collected broth can be separated into a bacterial cake and a broth supernatant by centrifugation. The liquid is then concentrated and dialyzed against tap water using a semipermeable membrane such as a cellophane tube to remove dialyzable impurities. next
The inner retentate containing BBM-1644 (after insoluble material has been removed) can be saturated with a salting-out reagent such as ammonium sulfate to precipitate BBM-1644 as a crude solid. This solid can be desalted by dissolving it in water and dialyzing it against tap water. Further purification of crude BBM-1644 can be carried out by conventional methods used for other acidic peptides. For example, an aqueous solution containing BBM-1644 can be adsorbed onto an ion exchanger such as DEAE-Sephadex, DEAE-cellulose, CM-Sephadex or CM-cellulose and eluted with a neutral salt solution. Sequential chromatographic steps are preferably used depending on the gradient concentration of the salt solution used as eluent. then refined
The aqueous fraction containing BBM-1644 is concentrated to dryness by freeze-drying or the like. Physicochemical Properties of BBM-1644 Upon lyophilization, BBM-1644 is isolated as an amorphous white powder. High pressure paper electrophoresis (PH8.6)
When tested at 4500 V in 0.05 M barbital buffer, BBM-1644 showed a positive response to the anode after 1 hour.
Travels as an acid that travels 8.7 cm. BBM-1644 does not exhibit an appreciable melting point, but gradually decomposes above 240°C. Soluble in water, but methanol, ethanol, acetone, ethyl acetate and n-
It is virtually insoluble in common organic solvents such as hexane. This antibiotic in 0.25% aqueous solution [α]
²⁶ _D = -75.6° optical rotation. As shown in Figure 2, the UV spectrum of BBM-1644 is 275nm.
(E ¹ % ₁ cm82) and 310 nm (E ¹ % ₁ cm46, shoulder). Shows nearly identical UV spectra in water and 0.01N HCl, but 0.01N
It shows only a single maximum at 285 nm (E ¹ % ₁ cm 8.9) in NaOH. Figure 1 shows the IR spectrum of BBM-1644 measured in KBr. This spectrum is NH
and the presence of OH groups (3300 to 2980 cm ^-1 ) and amide groups (1650 and 1540 cm ^-1 ). This antibiotic reacts positively with phorin-Lowry, xanthoprotein, Biuret and ninhydrin reagents and decolorizes potassium permanganate solutions. Negative for Anthrone and Sakaguchi reactions. Cephadex G-75 Ovalbumin (MW)
43,000), chymotrypsinogen (25,000) and ribonuclease A (13,700), BBM-1644 elutes immediately after chymotrypsinogen, so its molecular weight is 22,000.
It is thought to be nearby. Elemental analysis of BBM-1644 shows 46.60% carbon, 6.45% hydrogen, 13.34% nitrogen and 0.20% sulfur. The presence of 13 types of amino acids in the molecule of BBM-1644 was revealed by amino acid analysis as shown in Table 4. Basic amino acids such as lysine, histidine and arginine are not present in BBM-1644. Table 4 Amino acid composition of BBM-1644 Amino acid relative composition of amino acids Alanine 8.8 Aspartic acid 6.0 Half-cystine 1.0 Glutamic acid 5.7 Glycine 8.7 Isoleucine 2.3 Leucine 1.0 Phenylalanine 1.4 Proline 4.1 Sellin 1.6 Threonine 7.2 Tyrosine 0.6 Valine 11.1 * Leucine content Arbitrarily set to 1.0. BBM-1644 is fairly stable in the PH range of 2-9, but stability decreases rapidly outside this PH range. Aqueous solution of BBM-1644 at neutral pH, 50℃
It is stable for 2 hours at . Upon exposure to ultraviolet light, BBM-1644's antibiotic activity is lost within 20 minutes. The physical properties of BBM-1644 described above indicate that it is a member of the group of protein antitumor antibiotics that includes neocarzinostatin, macromomycin, and auromomycin. However, BBM−1644
can be distinguished from known protein antitumor antibiotics by their molecular weight, amino acid content and paper electrophoresis. Table 5 shows the paper electrophoretic mobilities of BBM-1644, neocarzinostatin, and macromomycin. Table 5 Paper electrophoresis ^* mobility (mm from loading point) BBM-1644 +87 Neocarzinostatin +31 Macromomycin -20 * 4500V, 1 hour; barbital buffer PH8.6 Neocarzinostatin antibiotic group is 275 as well as
Although it shows a UV maximum around 350 nm, BBM-1644
It has UV maxima around 275 and 310 nm. Recently , Biochem reported that the 350 nm maximum of neocarzinostatin antibiotics may be due to a non-protein chromophore that is essential for their biological activity.
Res. Commun ． 95 :1351–1356 (1980). It is noted that BBM-1644 has a different chromophore than the known neocarzinostatin antibiotic family. Biological properties of BBM-1644 The antibacterial activity of BBM-1644 was determined by serial 2-fold agar dilution method. Plain agar medium was used for Gram-positive and Gram-negative bacteria; normal agar medium containing 4% glycerol was used for acid-fast bacteria, and Saboro-agar medium was used for molds. The activity is expressed as minimum inhibitory concentration (MIC) in agar medium and the results are shown in Table 6 along with those for neocarzinostatin. BBM-1644 showed strong inhibitory activity against Gram-positive and acid-fast bacteria, but did not inhibit the growth of Gram-negative bacteria and molds.
Although the antibacterial spectrum of BBM-1644 is similar to neocarzinostatin, the intrinsic activity of BBM-1644 is stronger than the latter in some of the tested bacteria. [Table] The ability of BBM-1644 to induce prophages in lysogenic bacteria (ILB) was evaluated using the method of Rein et al. ( Nature 196 : 783~
784, 1962) using neocarzinostatin as a reference compound. Plaque counts were performed on agar plates containing test material (T) and control (C). A T/C ratio of plaque number greater than 3 was considered significant and ILB activity was expressed by the minimum inducing concentration of the test compound. As shown in Table 7, the IBL activity of BBM-1644 is similar to that observed with neocarzinostatin, with a minimum inducing concentration of 1 mcg/ml. Table 7 Induced ILB activity of lysogenic bacteria by BBM-1644 (T/C) ^* 100 10 1 0.1 mcg/ml BBM-1644 10.4 10.3 6.0 1.2 Neocarzinostatin 28.6 20.4 10.8 0.7 * Significant activity: T/C>3.0 Antitumor activity of BBM-1644 in lymphoid leukemia
p388 was determined in mice (BDF ₁ strain). Each mouse was inoculated intraperitoneally with 3×10 ⁵ tumor cells. Mice were given graded doses of antibiotics intraperitoneally 24 hours after tumor implantation. Treatments were performed on days 1, 4 and 7 (q3d x 3 schedules) or once daily for 9 consecutive days (qd 1→9 schedules). Neocarzinostatin was tested as a control anti-tumor agent for comparison and the results are summarized in Table 8.
BBM-1644 was highly active against murine leukemia in the dose range of 0.03-1.0 mg/Kg/day for both treatments. The antitumor activity of BBM-1644 is similar to that of neocarzinostatin at the lowest effective dose (qd 1→9
scheduler) or three times more powerful (q3d x 3 schedulers). [Table] Tatin [Table] The antitumor activity of BBM-1644 was also demonstrated in a second study against P388 leukemia in mice, the results of which are shown in Table 9 below. Details of the methods used in this study can be found in Cancer Chemother . Rep ．
3:1-87 (Part 3), 1972. [Table] [Table] Acute toxicity of BBM-1644 in mice (ddY lineage)
determined by a single intraperitoneal administration at
LD ₅₀ was calculated to be 5.8 mg/Kg. As shown above, BBM-1644 has strong antibacterial activity against Gram-positive and acid-fast bacteria and is thus useful in the treatment of mammals and other animals against infections caused by said bacteria. . Additionally, it can be used for other common applications of antimicrobial agents, such as disinfecting medical and dental instruments. The induction of prophages in lysogens and the significant antitumor activity shown against P388 leukemia in mice indicate that BBM-1644 may also be therapeutically useful in blocking tumor development in mammals. Accordingly, the present invention provides a method of treating an animal host affected by a bacterial infection or malignancy;
It is an effective antibacterial or tumor inhibiting amount for the host.
The method comprises administering BBM-1644 or a pharmaceutical composition thereof. In another aspect, the invention provides a pharmaceutical composition comprising an effective antimicrobial or tumor inhibiting amount of BBM-1644 in combination with an inert pharmaceutically acceptable carrier or diluent. These compositions may be comprised in any pharmaceutical form suitable for parenteral administration. Formulations according to the invention for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions or emulsions. They can also be prepared in the form of sterile solid compositions that can be dissolved in sterile water, saline, or some other sterile injectable medium immediately before use. The actual preferred amount of BBM-1644 used is
It will be appreciated that this will vary depending on the particular composition formulated, the mode of application and the particular location, host and disease being treated. Many factors modify the action of drugs, such as age, weight, sex, diet, time of administration,
route of administration, excretion rate, host conditions, drug combinations,
Reaction susceptibility and severity of the disease will be taken into account by the person skilled in the art. Administration can be carried out continuously or periodically within the maximum tolerated dose. The optimal rate of application for a given set of conditions can be ascertained by one skilled in the art using routine dosing tests according to the guidelines above. The following examples are presented for illustrative purposes only and are not intended to limit the scope of the invention. DEAE cellulose is diethylaminoethyl ion exchange cellulose. Cephadex G-50 is a supergel manufactured by Pharmacia Fine Chemicals, Inc. DEAE
Cephadex A-50 is a diethylaminoethyl anion exchange gel manufactured by Pharmacia Fine Chemicals, Inc. Safedex is Pharmacia
It is a trademark of Huain Chemicals, Inc. Example 1 Fermentation of BBM-1644 Using agar slants with well-attached growth of Actinomas sp. H710-49, 1% mannitol,
Contains 2% peptone and 1% yeast extract,
A seed medium (100 ml in a 500 ml Erlenmeyer flask) whose pH was adjusted to 7.2 before sterilization was inoculated. This seed culture was incubated for 72 hours at 32° C. on a rotary shaker (250 rpm) and 5 ml of this culture was transferred to a second seed medium (100 ml) having the same composition as the first seed medium. 1st
were cultured under the same conditions used for seed culture.
Using the thus prepared inoculum, 2.5% mannitol, 0.5% glucose, 1% soybean meal,
0.5% peptone, 1% meat extract, 0.3% _CaCO3 and
Fermentation was started in a 500 ml Erlenmeyer flask containing 100 ml of fermentation medium with a composition of 0.2% NaCl.
Fermentation was carried out at 28°C on a rotary shaker using a rotation of 250 rpm. Antibiotic production was monitored by paper disc-agar diffusion assay using Bacillus subtilis M45 (Rec ^- mutant) as the test organism. The antibiotic activity in the culture broth gradually increases as the fermentation progresses, reaching approximately 300 mcg/mcg/day after 6-7 days.
Reached ml. Example 2 The collected broth (18 liters) obtained from the fermentation of Example 1 was transferred to a shear press centrifuge (Kokusai No.
4A) to separate the bacterial cell cake and broth supernatant. Concentrate the solution below 40°C to 10% of the original volume.
The concentrate was dialyzed against tap water in a cold room over Cellophane Anti-Ubing (Union Carbide). Concentrate the inner retentate to approx.
The volume was reduced to 1.5 liters, and it was centrifuged (8000G) to remove insoluble matter. The clear supernatant was saturated with ammonium sulfate and left at 5°C for 5 hours. The resulting precipitate was collected by centrifugation, dissolved in water (300 ml), and desalted by dialysis against tap water. The dialysate (700 ml) had BBM-
Contained 22 g of 1644 crude solid. The remainder of the solution was used for the next purification without concentration to avoid decomposition. The antibiotic solution was passed through a column of DEAE-cellulose (ll ^- , 400 ml) and the column was soaked with water (1
The cells were washed with 1/15M phosphate buffer (PH7.0) containing 0.3M sodium chloride.
The active fractions were combined (300 ml), dialyzed for 18 hours against tap water, and chromatographed on a column of DEAE-cellulose (400 ml) equilibrated with 1/15M phosphate buffer (PH 7.5). The column was developed with the same buffer containing increasing amounts of sodium chloride (0-0.2M). Desalting the active eluate by dialysis;
It was loaded onto a column of DEAE-Sephadex A-50 (17 ml). gradient concentration of sodium chloride (0
~0.3M) in 1/15M phosphate buffer (PH
7.5) expanded this column. B. subtilis
pooling the active fractions determined by M45 quantification;
Dialysis was performed against running water for 18 hours. 1/15M phosphate buffer (PH7.0)-NaCl (0-0.3M) as eluent
DEAE A-50 (18
ml) was chromatographed on a column. Appropriate fractions were collected, concentrated to 10 ml and applied onto a Sephadex G-50 column for desalting. The column was eluted with deionized water and the active eluate was lyophilized to yield 120 mg of a white powder. thus obtained
The BBM-1644 sample was homogeneous as revealed by polyacrylamide gel electrophoresis. According to another feature of the invention: 1 an antibiotic in a recoverable amount when cultured in a normal aqueous medium containing assimilable carbon and nitrogen sources;
a biologically pure culture of the microorganism Actinomaszula sp. ATCC 39144 capable of producing BBM-1644;
3. A method for treating a host, which comprises administering BBM-1644 to an animal host affected by a malignant tumor that is sensitive to BBM-1644. Also provided are methods of treating a host, and 4. a pharmaceutical composition comprising an effective antimicrobial or tumor inhibiting amount of BBM-1644 in combination with a medical carrier or diluent.

[Brief explanation of drawings]

FIG. 1 of the accompanying drawings is an infrared absorption spectrum (KBr pellet) of BBM-1644 of the present invention.
FIG. 2 is the ultraviolet absorption spectrum of BBM-1644 of the present invention in water, 0.01N HCl and 0.01N NaOH.

Claims (1)

[Claims] 1. (a) Effective in inhibiting the growth of Gram-positive and acid-fast bacteria; (b) Effective in inhibiting the development of P388 leukemia in mice; (c) A lysogen. (d) soluble in water but practically insoluble in methanol, ethanol, acetone, ethyl acetate and n-hexane; (e) red at 3400-3200, 1640 and 1530 cm ^-1 The external absorption spectrum (KBr) is shown; (f) UV λ ^H20 _nax nm (E ¹ % ₁ cm): 286 (9.8) and 308 (7
.8)
(g) has an optical rotation of [α] ²⁶ _D -75.6° in a 0.25% aqueous solution; (h) has no sharp melting point, but above about 240°C Gradually decomposes; (i) migrates approximately 8.7 cm toward the anode during electrophoresis using 0.05 M barbital buffer at PH 8.6 for 1 hour at 4500 V; (j) Elemental analysis: C, 46.60%; H ,6.45%;N,
13.34%; S, 0.20% and O (by difference) 33.41%
(k) It is a high molecular weight peptide with a molecular weight of about 22,000; (l) Decolorization of potassium permanganate solution yields positive phorin-Lowry, xanthone protein, Biuret and ninhydrin reactions, and negative anthrone and ninhydrin reactions. shows the Sakaguchi reaction; and (m) hydrolysis produces alanine (8.8), aspartic acid (6.0), half-cystine (1.0), glutamic acid (5.7), glycine (based on the content of leucine arbitrarily taken as 1.0). 8.7), isoleucine (2.3), leucine (1.0), phenylalanine (1.4), proline (4.1), serine (1.6), threonine (7.2), tyrosine (0.6) and valine (11.1). Antibiotics
BBM−1644. 2. A BBM-1644-producing strain of Actinomadura sp. in a normal aqueous medium containing assimilable carbon and nitrogen sources under deep aeration conditions, such that a substantial amount of BBM-1644 is produced by the bacterium in the medium. A method for producing BBM-1644, an antibiotic characterized by culturing until 3. The method according to claim 2, in which BBM-1644 is recovered from the culture medium. 4 BBM-1644 producing bacteria is Actinomaszula sp. H710
49 (ATCC 39144). 5. An antibacterial agent consisting of an effective antibacterial amount of BBM-1644. 6. The antibacterial agent according to claim 5, which contains a pharmaceutical carrier or diluent. 7. An antitumor agent comprising an effective tumor-inhibiting amount of BBM-1644. 8. The antitumor agent according to claim 7, which contains a pharmaceutical carrier or diluent.