Novel antibiotic SF-2107 series substance and process for preparing the same

Disclosed is a novel antibiotic SF-2107 series substance consisting essentially of Substance SF-2107 A-1, Substance SF-2107 B and/or Substance SF-2107 C, each having the phisico-chemical properties as described in the specification, which substance exhibits an antibacterial activity against both gram-positive and gram-negative bacteria and is useful for pharmaceuticals, bactericides, disinfectants, etc. Also disclosed is a process for preparing the substance.

This invention relates to a novel antibiotic substance and a process for 
preparing the same. More particularly, it is concerned with a novel 
antibiotic SF-2107 series substance, and also with a process for preparing 
a novel antibiotic SF-2107 series substance which comprises cultivating an 
antibiotic SF-2107 series substance-producing microorganism belonging to 
the genus of Dactylosporangium on a culture medium and isolating said 
antibiotic SF-2107 series substance from the resulting cultured broth. 
In this application, the above-mentioned "an antibiotic SF-2107 series 
substance" means a Substance SF-2107 A-1, Substance SF-2107 B and/or a 
Substance SF-2107 C. 
The present inventors have found that there is produced in the cultured 
broth of a certain strain a substance which exhibits an antibacterial 
activity against both gram-positive and gram-negative bacteria. Then, they 
have isolated said active substance in a pure state and investigated its 
properties and, as a result, confirmed that it is a novel antibiotic 
substance different from known substances. This active substance has been 
named an antibiotic SF-2107 series substance, and more particularly, 
Substance SF-2107 -1, Substance SF-2107 B or Substance SF-2107 C. 
As the new antibiotic SF-2107 series substance-producing microorganism, 
there may be employed any of those having an ability to produce a 
sufficient amount of the SF-2107 series substance to be isolated in a 
cultured broth, but, as an example of these strains, there may be 
mentioned the SF-2107 strain that was first isolated by the present 
inventors from a sample of soil collected at Jorinji Temple in 
Komaoka-cho, Tsurumi-ku, Yokohama-shi, Kanagawa-ken, Japan. Morphological 
properties of this strain are as recited below. 
I. MORPHOLOGICAL CHARACTERISTICS 
The substrate mycelium is well-branched, wavy-elongated and of a diameter 
of about 0.5-0.6.mu.. No branched substrate mycelia are usually observed 
on either agar media or liquid media. 
Aerial mycelia are hardly observed and appear not to be substantially 
formed. The SF-2107 strain forms one or tufty sporangium over the surface 
of agar medium. Many sporangia can be observed on starch agar medium, 
glycerol. asparagene agar medium and the like. The sporangium is of a 
finger-shape and of a size of about 0.8-1.1.times.2.5-4.0.mu.. Each 
sporangium contains therein 3-4 spores in a line. When the surface portion 
of agar medium containing sporangia are scratched off, suspended in a 
sterile water, allowed to stand for not less than 30 minutes and then 
viewed under the microscope, it can be observed that the spores show an 
active motility. When such spores are viewed under the electron 
microscope, the spores are elliptical or short cylindrical and their 
surface is smooth and several flagella are observed on their one end. 
II. CULTURAL CHARACTERISTICS ON VARIOUS CULTURE MEDIA 
Cultural characteristics of the SF-2107 strain on various culture media are 
as shown in the following Table. As the standard for the symbols shown in 
the square bracket [] with respect to color indication was employed the 
color chip number as taught in "Color Harmony Manual", available from 
Container Corporation of America. Observation was effected after 
cultivation at 28.degree. C. for 14-21 days. 
______________________________________ 
Spor- Aerial Soluble 
Medium Growth and Color 
angium mycelium 
pigment 
______________________________________ 
Sucrose .multidot. 
Thin, very poor, 
Scant None None 
nitrate agar 
colorless 
Glucose .multidot. 
Very poor, Abun- " " 
asparagine 
colorless dant 
agar 
Glycerol .multidot. 
Poor, colorless 
Abun- " " 
asparagine dant 
agar 
Starch Poor, colorless to 
Abun- " " 
agar pale rose beige [4ec] 
dant 
Oatmeal Poor, colorless to 
Abun- " " 
agar very pale yellow 
dant 
Yeast malt 
Moderate, amber to 
None " " 
agar pale orange [3lc] 
Tyrosine Moderate, melon 
Abun- " " 
agar yellow [3ga.about.3ea] 
dant 
Nutrient agar 
Poor, colorless 
None " " 
BENNETT's 
Moderate, melon 
" " " 
agar yellow to amber 
[3ea] 
Malic acid 
Very poor, colorless 
Scant " " 
calcium agar 
______________________________________ 
III. PHYSIOLOGICAL PROPERTIES 
(1) Growth temperature range: Growth on a yeast.malt.agar medium at a 
temperature range of 20.degree.-42.degree. C., good growth at 
28.degree.-37.degree. C. 
(2) Liquefaction of gelatin: Negative (20.degree. C., cultivated for 21 
days) 
(3) Hydrolysis of starch: Negative (28.degree. C., cultivated for 14 days) 
(4) Nitrate reduction: Positive (28.degree. C., cultivated for 14 days) 
(5) Milk peptonization: Negative (28.degree. C., 37.degree. C., cultivated 
for 14 days) Milk coagulation: Negative (28.degree. C., 37.degree. C., 
cultivated for 14 days) 
(6) Halotolerance: Growth at 1.5%, but no growth at not less than 3.0% 
(7) Melanin formation: Negative 
______________________________________ 
IV. Carbon source utilization: 
Carbon source Growth 
______________________________________ 
D-glucose ++ 
D-xylose + 
D-fructose ++ 
D-mannitol + 
L-arabinose + 
L-rhamnose + 
i-inositol + 
sucrose ++ 
raffinose + 
glycerol + 
none + 
______________________________________ 
++ Good growth (utilization: +) 
+ Minor growth (utilization: -) 
The basal medium employed: 
______________________________________ 
Yeast extract (Difco Co., Ltd.): 
1 g 
Calcium carbonate: 0.2 g 
Agar (Difco Co., Ltd.): 15 g 
Distilled water: 1000 ml 
______________________________________ 
V. CELL WALL COMPOSITION 
As a result from analysis according to Becker et al method [See Appln. 
Microbiol., 13:236 (1965)], the diaminopimelic acid in the cell wall 
components was mainly of a hydroxy type. 
From the foregoing properties, SF-2107 strain has been identified as a 
strain belonging to the genus Dactylosporangium. 
The present inventors have named the SF-2107 strain as Dactylosporangium 
sp. SF-2107. 
This strain has been deposited with Fermentation Research Institute, Agency 
of Industrial Science & Technology, Ministry of International Trade & 
Industry, Japan and its accession number of an application in the 
Fermentation Research Institute is FERM No. 5351. The corresponding strain 
has also been deposited with American Type Culture Collection under 
accession number of ATCC No. 31744. The former was deposited on Apr. 1, 
1980 and the latter on Nov. 6, 1980. 
The SF-2107 strain is apt to have its variable properties as can be seen in 
the case of many strains in actinomycetes and may be variable by 
artificial variation procedures, for example, using an ultraviolet ray, an 
X-ray, a radiation, a chemical agent and the like. However, even any 
variants are usable in the present process which are capable of producing 
the SF-2107 series substance and belong to the genus of Dactylosporangium. 
In the process according to the present invention, the aforesaid strain can 
be cultivated on a culture medium containing those nutrients utilizable by 
ordinary microorganisms. As a nutrient source, there may be employed any 
materials hitherto well-known to be utilized for the cultivation of 
actinomycetes. For instance, there may be employed as a carbon source 
glucose, glycerol, sucrose, starch, dextrin, starch syrup, molasses, 
soybean oil etc. On the other hand, there may be employed as a nitrogen 
source soybean meal, wheat embryo, meat extract, peptone, yeast extract, 
dry yeast, corn steep liquor, cotton seed cake, fish meal, ammonium 
sulfate, sodium nitrate, urea etc. Additionally, there may be added 
inorganic salts such as calcium carbonate, sodium chloride, cobalt 
chloride, phosphates etc., if necessary, and, further, organic and 
inorganic substances may be suitably incorporated which can promote the 
growth of a strain and the production of the SF-2107 series substance. 
For cultivation, there may be any of cultivation methods under aerobic 
conditions similarly to the method for the production of general 
antibiotic substances, but submerged culture is most preferable. Suitable 
cultivation temperature may be 25.degree.-37.degree. C., but it is 
preferable in many instances to conduct the cultivation around 28.degree. 
C.-32.degree. C. Maximum accumulation can be accomplished in 3-10 days in 
the production of the SF-2107 series substance by either shaken culture or 
tank culture. 
In assay of the SF-2107 series substance, there is used a biological assay 
with Vibrio percolans ATCC 8461. According to this assay, the SF-2107 
series substance shows a linear relationship between its logarithmic 
concentration between its inhibition zone size at 1000 mcg/ml-31.3 mcg/ml, 
while inhibition circle diameters of 26-14 mm can be seen, respectively, 
according to a paper disc method. 
The SF-2107 series substance has the under-mentioned physico-chemical 
properties and, accordingly, can be extracted and purified upon such 
properties, but more effective extraction and purification is feasible 
according to the procedures as shown below. Namely, the active component 
which is involved mainly in a solid portion obtained after removal of a 
liquid portion from a cultured broth by filtration, can be extracted from 
the solid portion with aqueous acetone, aqueous methanol and the like and, 
after the organic solvent is distilled off, extracted with a solvent such 
as ethyl acetate and the like. In the case where the active component is 
also contained in the filtrate, it may be extracted from a culture 
filtrate with a solvent such as ethyl acetate and the like. Thereafter, 
the solvent, e.g., ethyl acetate layer containing the active component is 
concentrated to dryness and then a pure form of the SF-2107 series 
substance can be obtained by any suitable combination of chromatography 
using as a support silica gel, alumina, Sephadex LH-20 (Pharmacia Fine 
Chemicals), Florisil and so on and a counter-current distribution method. 
The SF-2107 series substance thus obtained can develop a single spot on 
all of thin-layer chromatography using various solvent systems and thus 
can be regarded as a pure form. 
Physico-chemical properties of the SF-2107 series substance obtained 
according to the above-mentioned procedures are as follows; FIGS. 1, 3 and 
5 referred to therein show each an ultraviolet absorption spectrum of the 
Substance SF-2107 A-1, Substance SF-2107 B and Substance SF-2107 C, 
respectively, as measured with a methanolic solution of 25 mcg/ml; FIGS. 
2, 4 and 6 also referred to therein show each an infrared absorption 
spectrum of the Substance SF-2107 A-1, Substance SF-2107 B and Substance 
SF-2107 C, respectively, as measured in a potassium bromide tablet: 
I. SUBSTANCE SF-2107 A-1 
Elementary analysis: 
C, 58.39 wt %; H, 7.88 wt %; O, 33.73 wt % (balance) which does not 
contain any nitrogen, sulfur, phosphorus, or halogen. 
Molecular weight: 
900-1100 (gel filtration method) 
Melting point: 
170.degree.-184.degree. C. (slowly molten) 
Specific rotation: 
[.alpha.].sub.D.sup.25 =+20.degree. (c=0.2, Methanol) 
Ultraviolet absorption spectrum: 
Absorption maxima at 250 nm (E.sub.1 cm.sup.1% =72), 342 nm (206), (in 
methanol) 
Infrared absorption spectrum: 
Characteristic absorption bands at 3460, 2950, 1730, 1630, 1610, 1460, 
1380, 1300, 1280, 1130, 1100, 1040, 910, 860, 820, 790, 750, 740, 690 
cm.sup.-1 (KBr tablet method) 
Color reaction: 
Iodine reaction, Lemieux reaction, positive Ninhydrin reaction, ferric 
chloride reaction, negative 
State: 
Pale yellow powder 
Neutrality, acidity or basicity: 
Acting as a neutral or weakly acidic substance (electrophoresis) 
Silica gel thin-layer chromatography: 
Rf=0.67 (chloroform:methanol=5:1)=0.56 (acetone:benzene=5:1) 
Solubility: 
Soluble in methanol, acetone. Sparingly soluble in benzene, chloroform, 
n-hexane, water 
II. SUBSTANCE SF-2107 B 
Elementary analysis: 
C, 53.66 wt %; H, 6.75 wt %; O, 39.59 wt % (balance) which does not 
contain any nitrogen, sulfur, phosphorus, or halogen 
Molecular weight: 
900-1100 (gel filtration method) 
Melting point: 
170.degree.-175.degree. C. (slowly molten) 
Specific rotation: 
[.alpha.].sub.D .sup.23 =-5.degree. (c=1, Methanol) 
Ultraviolet absorption spectrum: 
Absorption maxima at 242 nm (E.sub.1 cm.sup.1% =52), 343 (206), (in 
methanol) 
Infrared absorption spectrum: 
Characteristic absorption bands at 3450, 2970, 2930, 1720, 1600, 1440, 
1370, 1270, 1170, 1130, 1040, 980, 790, 750, 690 cm.sup.-1 (KBr tablet 
method) 
Color reaction: 
Iodine reaction, Lemieux reaction, positive Ninhydrin reaction, ferric 
chloride reaction, negative 
State: 
Slightly yellowish powder 
Neutrality, acidity or basicity: 
Acting as a neutral or weakly acidic substance (electrophoresis) 
Silica gel thin-layer chromatography: 
Rf=0.34 (chloroform:methanol=5:1)=0.51 (acetone:benzene=5:1) 
Solubility: 
Soluble in methanol, acetone. Sparingly soluble in benzene, chloroform, 
n-hexane, water 
III. SUBSTANCE SF-2107 C 
Elementary analysis: 
C, 57.50 wt %; H, 7.04 wt %; O, 35.46 wt % (balance) which does not 
contain any nitrogen, sulfur, phosphorus, or halogen. 
Molecular weight: 
900-1100 (gel filtration method) 
Melting point: 
155.degree.-168.degree. C. (slowly molten) 
Specific rotation: 
[.alpha.].sub.D.sup.23 =+29.8.degree. (c=1, methanol) 
Ultraviolet absorption spectrum: 
Absorption maximum at 345 nm (E.sub.1 cm.sup.1 %=258) (in methanol) 
Infrared absorption spectrum: 
Characteristic absorption bands at 3430, 2930, 1720, 1620, 1600, 1440, 
1370, 1300, 1270, 1130, 1100, 1050, 900, 790, 750, 690 cm.sup.-1 (KBr 
tablet method) 
Color reaction: 
Iodine reaction, Lemieux reaction, positive Ninhydrin reaction, ferric 
chloride reaction, negative 
State: 
Slightly yellowish powder 
Neutrality, acidity or basicity: 
Acting as a neutral or weakly acidic substance (electrophoresis) 
Silica gel thin-layer chromatography: 
Rf=0.27 (chloroform:methanol=5:1)=0.07 (acetone:benzene=5:1) 
Solubility: 
Soluble in methanol, acetone. Sparingly soluble in benzene, chloroform, 
n-hexane, water 
Minimum inhibitory concentrations (MIC) against various bacteria of the 
SF-2107 series substance as assayed by an agar dilution method are given 
in the following Table 1, which demonstrates effectiveness against both 
gram-positive and gram-negative bacteria. Also, acute toxicity tests of 
the present substance in mice showed that all animals survived at 90 mg/kg 
via intraperitoneal administration. Accordingly, the antibiotic SF-2107 
series substance is useful for pharmaceuticals, drugs for animals, 
bactericides, and disinfectants, as well as for convertion materials 
thereto. 
As the pharmaceuticals for which the antibiotic SF-2107 series substance or 
a salt thereof is useful, there may be mentioned various types for oral, 
topical or parenteral administrations such as tablets, capsules, creams, 
syrup, suspensions, solutions, powders and sterilized compositions 
suitable for injections or the like. The pharmaceuticals according to the 
present invention may be administered at the dosage of 30 to 50 mg a day, 
more generally, 3 to 300 mg a day. 
Comparison was effected in physico-chemical properties and biological 
activities between the SF-2107 series substance and known antibiotic 
substances, by which it has been proven that there are no corresponding 
known substances, and, therefore, the present substance has been evident 
to be a novel antibiotic substance. 
Examples are given below for the production of the SF-2107 series 
substance, but it is to be noted that many other variation and 
modification means not illustrated herein may be applied. 
TABLE 1 
______________________________________ 
MIC (mcg/ml) 
SF-2107 
Test Organism A-1 B C 
______________________________________ 
Staphyloccus aureus JC-1 
3.13 25 0.78 
Staphylococcus epidermidis 
3.13 50 6.25 
ATCC 14900 
Bacillus anthracis No 119 
0.78 12.5 0.39 
Escherichia coli JC-2 
&gt;100 &gt;100 &gt;100 
Escherichia coli RGN 823 
0.78 50 6.25 
Salmonella typhi D-901-W 
25 &gt;100 &gt;100 
Klebsiella pneumoniae 
&gt;100 &gt;100 &gt;100 
PCI 602 
Proteus vulgaris OX 19 
6.25 50 6.25 
Serratia marcescens 
25 &gt;100 &gt;100 
MB-3838 
Pseudomonas cepacia 
3.13 50 6.25 
M-0527 
Pseudomonas maltophilia 
0.20 100 6.25 
M-0627 
______________________________________ 
Midium: Heart infusion agar (Eiken Chemical Ltd.)

EXAMPLE 1 
As a seed culture, there was used Dactylosporangium sp. SF-2107 strain 
(FERM. No. 5351 or ATCC No. 31744) and, as a seed culture medium, a medium 
containing glucose 1.0%, soluble starch 1.0%, Polypepton 0.5%, meat 
extract 0.2%, yeast extract 0.3%, soybean meal 0.2% and calcium carbonate 
0.2% (pH 7.0 before sterilization). 
Six to 7 platinum loops of the seed culture, which had been cultivated on a 
yeast-malt-agar slant medium at 28.degree. C. for 14 days, were inoculated 
into 20 ml of the above-mentioned seed culture medium in a 100 ml volume 
Erlenmeyer flask and then shaken culture was effected at 28.degree. C. for 
6 days. This was used as the first seed culture and three flasks were 
cultivated. 
Subsequently, this seed culture was inoculated into 80 ml of the seed 
culture medium in each of six 500 ml volume Erlenmeyer flasks in 8 ml 
portions and then shaken culture was effected at 28.degree. C. for 3 days. 
This was used as the second seed culture. 
In each of the one hundred 500 ml volume Erlenmeyer flasks were placed 80 
ml of a production medium, into which the above-mentioned second seed 
culture was then inoculated at a rate of 5%. 
As the production medium, there was employed a culture medium having the 
composition of glucose 2.5%, wheat embryo 2.0%, Sungrain (manufactured by 
Suntory Ltd.) 0.5%, and sodium chloride 0.25% (pH 7.0 before 
sterilization). 
Cultivation was effected by shaken culture at 28.degree. C. for 7 days by 
means of a rotary shaker (220 rpm). After completion of the cultivation, 
filtration was done to remove the filtrate, 6 l of 80% acetone in water 
were added to the resulting solid and then stirring was effected to 
extract the Substance SF-2107 A-1. The acetone was distilled off under 
reduced pressure from the extract, the residue was dissolved in water to 
1.3 l of an aqueous solution, the pH of the resulting solution adjusted to 
9 and extraction was made twice with 1 l portions of ethyl acetate. The 
extracts were combined, concentrated to dryness under reduced pressure to 
afford 300 mg of an oily substance. This was dissolved in 3 ml of 
methanol, applied to a column packed with 100 ml of Sephadex LH-20 
(Farmacia Fine Chemicals) and developed with methanol to separate active 
fractions, which were then concentrated under reduced pressure and dried 
to give 120 mg of a powdery substance. The resulting powdery substance was 
applied to a column packed with 20 ml of Wako Gel C-200 (Wako Pure 
Chemical Industries Ltd.), which was developed with a mixed solvent of 
chloroform-methanol (50:1). Of the active fractions, those fractions 
showing a single spot in a thin-layer chromatography were concentrated to 
dryness to afford 16 mg of a pale yellow powder of the Substance SF-2107 
A-1. 
EXAMPLE 2 
As a seed culture, there was used Dactylosporangium sp. SF-2107 strain 
(FERM No. 5351 or ATCC No. 31744) and, as a seed culture medium, a medium 
containing soluble starch 2.0%, glucose 1.0%, wheat embryo 0.6%, soybean 
meal 0.2%, Polypepton 0.5%, yeast extract 0.3%, meat extract 0.2% and 
calcium carbonate 0.1% (pH 7.0 before sterilization). 
Five platinum loops of the seed culture, which had been cultivated on a 
yeast-malt-agar slant medium at 28.degree. C. for 14 days, were inoculated 
into 20 ml of the above-mentioned seed culture medium in a 100 ml volume 
Erlenmeyer flask and then shaken culture was effected at 32.degree. C. for 
96 hours. This was used as the first seed culture. Subsequently, this seed 
culture broth was inoculated in 8 ml portions into 80 ml of the seed 
culture medium in each of ten 500 ml volume Erlenmeyer flasks and then 
shaken culture was effected at 32.degree. C. for 72 hours. This was used 
as the second seed culture. 
In a 30 l volume jar fermenter were placed 20 l of a production medium and 
800 ml of the above-mentioned second seed culture were inoculated 
thereinto. As the production medium, there was used a medium having the 
composition of glucose 1.7%, sucrose 1.5%, wheat embryo 2.0%, yeast 
extract 0.2%, gluten meal 0.3%, and sodium chloride 0.25% (pH 7.0 before 
sterilization). 
Cultivation was effected by aerated agitation culture at 28.degree. C. for 
164 hours. After completion of the cultivation, filtration was done to 
remove the filtrate, 12 l of 80% acetone in water were added to a solid 
and stirring was made to extract an active ingredient. The acetone was 
distilled off under reduced pressure, the residue dissolved in water to 2 
l of an aqueous solution, which was then adjusted to pH 9 and extracted 
twice with 1.5 l portions of ethyl acetate. The extracts were combined and 
concentrated to dryness under reduced pressure to yield 800 mg of an oily 
substance. This was dissolved in 5 ml of methanol and applied to a column 
packed with 500 ml of Sephadex LH-20 (Pharmacia Fine Chemicals), which was 
then developed with methanol to separate active fractions. These fractions 
were concentrated to dryness to afford 280 mg of a powdery substance. 
This powdery substance was applied to a column packed with 100 ml of Wako 
Gel C-200 (Wako Pure Chemical Industries Ltd.), which was developed 
stepwise, first with 1000 ml of a mixed solvent of chloroform-methanol 
(25:1) and then with 2000 ml of a mixed solvent of chloroform-methanol 
(15:1) to collect 20 ml each of fractions by using a fraction collector. 
Active fractions were found at Fraction Nos. 41-65, Nos. 76-89 and Nos. 
95-120. 
The active fractions thus obtained were subjected to silica gel thin-layer 
chromatography using the solvent system (I) of chloroform-methanol (5:1) 
and the solvent system (II) of acetone-benzene (5:1). The fraction showing 
RF=0.67 with the system (I) and the fraction showing RF=0.56 with the 
system (II) were combined and concentrated to dryness under reduced 
pressure to give 60 mg of powdery Substance SF-2107 A-1. The fraction 
showing RF=0.34 with the system (I) and the fraction of Rf=0.51 with the 
system (II) were combined and concentrated to dryness under reduced 
pressure to give 52 mg of Substance SF-2107 B. Further, the fraction 
showing Rf=0.27 with the system (I) and the fraction showing Rf=0.07 with 
the system (II) were combined and concentrated to dryness under reduced 
pressure to give 93 mg of Substance SF-2107 C. 
EXAMPLE 2-1 
60 mg of the powdery Substance SF-2107 A-1 obtained according to Example 2 
was dissolved in methanol to a concentration of about 2% and fractionated 
by a high performance liquid chromatography, using a column of 
Micropondapak C 18 (Waters Co.) and, as an eluant, a mixed solvent of 
methanol-acetonitrile-water (7:1:3). The resulting fractions were 
concentrated to dryness under reduced pressure to give 36 mg of a pale 
yellow powder of pure substance of the Substance SF-2107 A-1. 
EXAMPLE 2-2 
52 mg of the Substance SF-2107 B and 93 mg of the Substance SF-2107 C 
obtained according to Example 2 were respectively dissolved in a small 
amount of methanol, and applied separately to a column packed with 50 ml 
of Sephadex LH-20 (Pharmacia Fine Chemicals), which were then developed 
with methanol. The active fractions were subjected to thin-layer 
chromatography using the aforementioned two solvent systems, and the 
fractions showing a single spot therein were combined and concentrated to 
dryness under reduced pressure to yield 38 mg of pure substance of the 
Substance SF-2107 B and 76 mg of pure substance of the Substance SF-2107 
C, respectively. 
EXPERIMENT 1 
Therapeutic effect of SF-2107 series substance on mice infected with 
Pseudomonas cepacia 
(a) Sample: Substance SF-2107 A-1 
(b) Host animals: ddY-SLC female mice of average body weight of 19 g of 
4-week age, each group consisting of 3 animals. 
(c) Test strain: Pseudomonas cepacia M-0527. The strain was cultured to 
prepare solutions containing the Pseudomonas cepacia. 
(d) Dose and administration of the sample: The samples were dissolved in 
ethanol and adjusted with sterilized physiological saline solution to form 
solution of given concentrations (ethanol concentration: 10%), which were 
administered orally and subcutaneously at 2 dose levels, respectively: 10 
mg/mouse and 1 mg/mouse in oral administration; 5 mg/mouse and 1 mg/mouse 
in subcutaneous administration. The dose volumes were 0.5 ml/mouse in the 
oral administration and 0.2 ml/mouse in the subcutaneous administration. 
(e) Test procedures: Solutions containing the Pseudomonas cepacia were 
inoculated into abdominal cavities of the respective mice (inoculated 
bacterial number: 6.75.times.10.sup.7 cells/mouse, MLD). Immediately 
thereafter, the sample solutions were administered orally, or 
subcutaneously at the femurs, at the dosage as prescribed above to observe 
the living bodies after 7 days. 
(f) Test results: As seen from the following Table, the Substance SF-2107 
A-1 showed excellent therapeutic effect on mice infected with Pseudomonas 
cepacia. 
______________________________________ 
Living bodies after 7 days 
Oral admin- 
Subcutaneous 
istration administration 
Control 
______________________________________ 
Dose 10 1 5 1 
(mg/mouse) 
Living 3 2 3 2 0 
bodies 
______________________________________ 
(3 mice for each group)?