LU84930A1 - ANTITUMOR ANTIBIOTIC - Google Patents

Description

»Λ»

DESCRIPTIVE MEMORY

FILED IN SUPPORT OF A REQUEST

□ ε

PATENT

FORMED BY

BRISTOL-MYERS COMPANY, for

Antitumor antibiotic.

"

The present invention relates to a new anti-tumor antibiotic / as well as its production and isolation.

The anti-tumor antibiotic which is the subject of the invention, also known as BBM-1644, is a new CD.YD.MdC.8 - 1 - SY-1711 "substance belonging to anti-tumor protein antibiotics, examples of which are neocarzinostatin, macromomycin and auromomycin.

Neocarzinostatin (also known as zinostatin) is an acidic protein macromolecule with a molecular weight of 10,700 consisting of a single polypeptide chain of 109 amino acids joined by two disulfide bridges. The production of the. neocarzinostatin by fermentation of different strains of Streptomyces carzinostaticus var. neocarzinostaticus is described in U.S. Patent No. 3,334,022 and in Antibiotics 18: 68-76 (1965). The amino acid sequence in neocarzinostatin has been described in Cancer Treatment Reviews _6, 239-249 (1979).

Macromomycin is a neutral or weakly acidic polypeptide with a molecular weight of about 15,000. The production of macromomycin by fermentation of Streptomyces macromomyceticus is described in United States Patent No. 3,595,954 and in Antibiotics 21: 44-49 (1968). The purification of macromomycin and the characterization of the purified compound are described in Antibiotics 29: 415-423 (1976).

Auromomycin is a weakly acidic polypeptide with a molecular weight of about 12,500 and an isoelectric point of pH 5.4. It consists of 16 different friend-noacids. The isolation of auromomycin from the culture broth of Streptomyces macromomyceticus and the characterization of the purified product are described in Antibiotics 32: 330-339 (1979).

BBM-1644 can be distinguished from known polypeptide anti-tumor antibiotics such as neocarzinostatin, macromomycin and auromomycin by its physicochemical properties, such as molecular weight, amino acid composition and elect. CD.YD.MdC .8 - 2 - SY-1711 * trophoresis on paper.

The present invention relates to a new protein anti-tumor antibiotic called BBM-1644 which is prepared by culture of a new strain of Acti-nomadura called Actinomadura sp. strain H717-49 (ATCC 39144) in an aqueous nutritive medium containing assimilable sources of carbon and nitrogen, in aerobic * biosis in submersion until production of a significant quantity of BBM-1644 by this microorganism in this culture medium and optionally by isolating BBM-1644 from the culture medium. The invention also relates to the antibiotic BBM-1644 in dilute solution, in the state of crude concentrate, in the state of crude solid and in the state of purified solid.

• In the drawings:

Fig. 1 shows the infrared spectrum of BBM-1644 (KBr pellet), and

Fig. 2 shows the ultraviolet absorption spectra of BBM-1644 in water, 0.01 N HCl and 0.01 N NaOH

The subject of the invention is a new protein anti-tumor antibiotic called BBM-1644 and its preparation by fermentation of a new strain; d * Actinomadura called Actinomadura sp. strain H710-49.

The above organism has been isolated from a sample of - * soil collected from the Federal Republic of Germany. A biologically pure culture of this organism has been deposited in the American Type Culture Collection, Washington, D.C., (United States of America) under No. ATCC 39144 in its permanent collection of microorganisms.

BBM-1644 inhibits the growth of various gram-positive and acid-resistant bacteria. This antibiotic also tends to confer certain phage properties on lysogenic bacteria and CD.YD.MdC.8 - 3 - SY-1711 * · inhibit the growth of lymphatic and solid tumors such as P388 leukemia in mice. The new antibiotic can therefore be administered as an antibacterial agent or as an anti-tumor agent to inhibit tumors in mammals.

Actinomycete strain H710-49 was isolated from a soil sample and prepared according to standard techniques in the form of a biologically pure culture for characterization. The H710-49 strain forms substrate mycelium and aerial mycelium. The substrate mycelium is long, branched and not fragmented into short filaments. Short chains of spores are carried at the end or branching of the aerial mycelium. Spore chains have 2 to 15 spores assembled into a chain (most often 4 to 8 spores) and are straight, angular or curly. The spore lees have a warty surface and are oval to elliptical in shape (0.5 ^ 0.6 x 0.7 x 1/2 / µm) with a round or pointed tip. Ripe spores are often separated by empty hyphae. Terminal turgor of the hyphae is occasionally observed on the substrate mycelium in Czapek agar and Bennett agar. Mobile spores, sporangia or sclerotic granules were not observed on any medium.

Unlike ordinary species belonging to the genus Streptomyces, the strain H710-49 grows slowly and forms an aerial mycelium which is not abundant in chemically defined media and natural organic media. The color of the aerial mycelium is white, which turns pink after sporulation in oatmeal agar, starch agar with inorganic salts and asparagine agar with glycerol. The substrate mycelium is in colorless, yellow mass, CD.YD.MdC.8 - 4 - SY-1711 * reddish brown or dark greyish brown. A melanoid pigment does not form, but a citrus yellow diffusible pigment is observable on asparagine agar with glycerol, on tyrosine agar and on Pridham-Gottlieb base agar with one or the other. source of glycerol, L-arabinose, D-xylose, L-rhamnose, D-glucose, D-fructose, trehalose and D-mannitol. The H710-49 strain grows at 20 ° C, 28 ° C and 37 ° C, but not at 10 ° C or 41 ° C. It is sensitive to 10% NaCl, but not 7% and resistant to 0.001% lysozyme. D-galactose, D-mannose, sucrose, raffinose and inositol are not used by the strain. The culture characteristics and physiological characteristics of the strain H710-49 are collated in Tables I and II respectively. Table III shows the profile of use of carbon sources by the strain.

CD.YD.MdC.8 - 5 - SY-1711

CNJ

Γ- fl)

* '0) G

* 0 4-1

* 0) C <G

^ in oc - on c "H g

0) r- ρ G

"<0 - 'ί-ι» QJ -I — i 3 o in <d · a) cm G 0) H Q) (0 œ

^ G 'd) <G G G O

O 4J O. 3 O G

OC G

O d) * 0) τη α> - · (0

in r-t E in - G

tp in G -G "G -" I -H g vd an O ·. Cil} -t o- 'CN -nvo ^

H d) O CN * - d) CM CO

t- Ό) c - en v <D * G - 'co K Go 0) <o u G -' * • 0) G G vo G Ό) * u Φ Ό G CN Q) G Ό <n G +>

fi 0+) rt * - G i — i OCOröG

U £ 0) 'n -P £! g rri G

P O <(0 fi r-i 0 'PO' PC C ~ - 'PO · ~ l C0 * Q) G 3 in d) H - · Η

(Ü4-) 0) ι — I d) G 1Γ) G Q) “0 O) r-i G

μ “(Ö G G G fi G -π— ιϋ G a G“ o fi M H OQ) O ü G U U ^ oë r-i “144 Οο'φ -P ο α> ο tu

Ο) · Η -H (1) O fl) G -H C O to G · Η C-rl J) C

Ό Ό Ό G U G G Ό 10 GO ni 'O G Ό Ή G

^ "Φ Ό)" i G Sf'O)> J) ri O h '5) "0) (d" e) <(C rc ^ 0) £ œ G -GGCE, 0 4-> GE τη E ft -not

"* G

M G

w 4) *

^ G U U P C P U Pi <P U (¾ <P

a Ό

ai O O

^ Ό G

t, O

^ C0 4-5 d) 1¾ G> t

01 P

• H +1

4J I

to 0) H

: -HP O) O

G G <1> C0 G

'0)> CO O' 0)

-P fl) O O O

ü H G G> 1

’(0 (0 i — I i — I

G <D fi Cn in

(ΟΌΟ I I

U ü 0) O

fi G G G

• H CD · Η Ή (ö i -— in Cn

G 0) Λί G G

• P 4-> fl) G G

X iC Φ G G

d) G G ft Dî - -P N 10 to

H -H L) G G

VS

rp - <l) r- | r-l H GO - GG 'P' P in O o Q) o

H G d) CD d) d) G

H CO O CO 10

-G P o «H O OP

3 C0 H 'fl) H H CO

O H '0) O' 0) 'd) H

C0> - θ'— Ü CD - CD.YD.MdC.6 -6- SY-1711 σ "- 'ΟΊ φ οο φ a) uk U +> Φ +3 -Ρ" ΰ φ k - <Φ < φ φ η 4-ι σι ιο β σ <Φ <rö m Ο 0 0 Di β ^ k - Φ Ο 0 • η k φ φ 'Φ Ο β * π Ο ß * -> Φ C 0 C (0 in to 0 φ β Ο η • CO Ο k · Γ-ι 0 Μ-ι fi - k fi k Ίϋ fi β 'Φ σ' Φ * ιϋ * ΐο 0 ^ ο ^ * Φ - 'β -—-— I β ^ οο γο η ο οσ> 'φνο .-- νο σ νο ιΗ φΜ4 ^ - mc \ - φ cn cn * φ - α Γ ^ νφ' - ^ ^ .φ <Τι '—- - »^ trj k Φ» φ CO k '-'tn k * φ β to φ ο - »φ u CO CD -k ü Φ Ό 00 k ß Ό Φ β COfNfO β fi Ο Φ k +> Φ φ O k Φ - i — i Φ

Ο £ · σ <φ H to Ë 4-1 H U H

~ 0 Ο β -, Ο β <φ, Ω "0) 0, Ο Φ ο 'ίϋ' tö ß p co Φ" · φ ß r- φβφ 4-> 10 Φ - 'k ^ ·. 4-) 0 Γ "-» βφ> • Η φ φ Η Η rQ -'- 'φβ φ Φ · - φ Ο 1-4 · Η - φ φ φ 0 π3 k krû ^ Φ kH kr-> km Ml, Q Η kkk to r4 0 0 ^ "* Φ * Φ 'Φ OU MD U -Φ OUO ü - Oh - 4-> k σ k Ο Φ Ouk04-> k®- OHO 4-> Φ -r4 Ο Φ Φ ß -Φ β -Φ · ι4 β -k β -φ -Η β -φ β φ β 4 0 · 4 β Ό Ό Ο k Η Φ Ό ιϋΌΌ 0 Ό ιίΌΌ Ιϋ Ό 0k0 Ό ΟΌ ιϋ Η »Φ β (0 < Φ -Φ Ο k Ο -Φ φ> Φ4 Ο'Φ'Φ Ο Φ Φ k -Φ β -Φ -φ φ H-Hkftß ΕΟΕΕ · π Ε, Ω Ë £ ß Ë -η k, Ω Ε -k Ë ß k 0

D 4J

rH

laughed! 0 U Pi laughed! O U Pi laughed! O U Pi laughed! O OPiri! Q UPiriJQ

ü

H

Φ PI Ό - co ω to

Φ 10 O

laughed! 0 Φ 4J β

Cf 0 -k

Eh -k Ü1 IÖ Ck

4J -k -> k M

10 β r- 4-> H

• k Φ X ^ * j-ι σ ο φ -Φ k ß Φ 4-) O 4-> ß ü ß H -k

Φ -k tQ Φ O

k H Ë> Φ to ^ Φ UH Φ Φ Φ Ό Ό

to β CM

I -k 4-> Φ ß tO -k ο β Ο Ο Φ Φ β -k ^ k> k k -k> 1 -k 4J Ο, φ Ë -P 4-) X CO H4

Φ -k Φ H

Φ k - w Φ

1 H «- H -P HH

0 Φ • Φ 'Φ β * Φ k * Φ ο 0 φ β φ φ φ> Φ 01 (0 01 ΟΙ φ 01 0¾ Ο Ο Ο Η Ο Η C0 Η Η Η Η -φ η' Φ 'Φ' Φ Φ 'Φ

CD —- CD CD CD Ό CD

CD.YD.MdC.8 -> 7 - SY-1711 Φ Ul - φ ε -p ·

r-f P P U

Æ o m ·

• H C £ Q

03 U

30

^ -P d) C

0) IM g O

G P · Ρ, _, fl + J

P -P - tfi "0 c CP

P <(Ü Γ- I C

Λ CO f "+ J» J_, -H

<T1 ^ g 1 o £ '(0 i-l dl P H ίο

Cn »Φ Ë" ο id ^ P tJiO ok O c '0) * h 00 3' S & S ΓΛ - - 'PO o ® π £ EH ü S in σ û) rr-. Ym ^ pn in wd ^ < d P +) Λ l + i vo P npc H.

O <nj · - CM ~ VD -P ·> ο ^ n

| H tOCN “- O CM <(C G W 33 P

r * · -p vd 4 (u p ^ C φ .j ffl P ~ 0 P ~ P * P ω u

Cn G MD U rd P φ "Q)" Φ rd ’C φ d · ι — i MD m S.

£ 1) U ri 0 pic id, φ * 2 p

U p G £ E -P> -ri C ri g I

- P -PO <1ϋ fi P E + j ^ n φ ο <(β * P · Φ ü) · - P Dr; o P ω G Φ> -h cm * £ * p φ · -H ΠΦΡ-ΗΦΡΌΟ) '-IpCCn, 3 φ Φ id p rd rri p P '10 MD fl · n tn rri »φ-π + JpOU ü ü QO u

P <fß ΟΦ'ΦΟ-Ρ ÎP

φ “φ c” œ c · η c u · η c u E jj Ό 13 D · Ρ MD D Ό 3 ßr ß 0 2φο H OPPPP MD rd OMDMD COIIJI m Φ g £ tn-P £ E -nin Ë C w £ g

3 billion ^ "J

not a word ·% . · · H ß! I1 Jj ^ ra

Mj P üK <fl ÜPÎ <! D O P 3 £ Ο · Η Φ O1 K £ H -P> 7 *> i φ φ ro Φ P f-i · £ · σ C £ P'®H “o p g φ p

fflra -P a 3 O

Φ & fit C: ri

<P Φ -p O

a1 a - p p m u

Eri -H I flfifiri -μ Φ Φ Φ fl Ό to P IQ rri rri T5 · Η -H P Φ 3 “o

P> C O> P

* φ Φ -h O x -P

P H “fl * ® g o § il“ p c m φ φ φ 5 u p -o m es j, o id a *

U £ ro · 'φ -P

-H φ φ -H

(Ü CD O g 3 £ P / φ 3 O Cri + J ri-> P <0 "P · Η Ό = 4j i"! Ûitc-Ρρφιη Φ Φ (tw'ô + '+ Jr' GI · η P md rd σ CP 'ΦΟΒΕΡΉ

Φ φ> p r * -p -P

CQ Mri P ü O tmi.

r- "Φ U ο P · φ DVO» II ri ri>

Ό (I) £ “OH O

o O U H ü -P g

Φ Φ C

w m o o eu rri rri CO * MD MD H * * Ü o ^ * * * ÇD.YD.MdC.8 - 8 - SY-1711 d

Ο CD

Ο Ό 'Cd Μ “CD -ri 3 3 g> Ε CD CD (Ο>

~ (Ο (Ο -C CO CD

"DD - Ο CD rri

’J CD tri + J Η CD

ρ d -ri MD O CD

C -r-l (U ^ (0 Λ D'H Ό + J -P CD O P '0)

ir-p îcocuü -p * cji + J

Φ CD rri U CO “H X CD -ri C 'CD 3 O -H CD C -P (0

2 d Cnn h Q - 0 cd P

ZL CD 'CD H + J +>

~ ffi (0 CJi 'CD Or CD X

"d H + j g» cd> i d cd <d

<3 \ 5 CD CD CD MD P -ri C

-φ · TJO '<d> C P d -P CO υ O

I ^ (D Ο Ο I O <D -P

O £ Q> 3 d CD 'CD rl CD D> û rH ”CD CD O 3 H 14> 1 SS d) t" o -ri +> σ * -p -Hd-Pft

îü H ri ft IÜ -H 3> P

'CD -h il) h rd o <U rd <L> 4J

<D Ü 2 && 4ril ffl ri ri Ή (D

VS

O

3 o

CO CJ CD

o U U

cd oo »d rri NO li

H CN CD CD

H CD 'CÖ CD 4-! -P

Ό 'Cd -ri CD'CD

CD Ο Ή CD I — I CD P d

D CD cd MD U O E

^ 3 E P -ri O

C O * "-ri'CD CD · 4J 0 -ri h ÎCCOU (β H Cn n ^ 0 ° 1-10 O SEEiQHd 3 0 J H ^ cd ^ O Cn-ri

0 Jii CD CD Oi -ri cd-P

m -ri Kou-p +> <DOcd

CD d d CD U CD U CD

ÔÎ> 1 cd cd U Cti> 1 -ri

Jd CD CD ° U * H H CD d

Eh Oj CDCDC ^ -o ”(]) ΟΌΟ-Ρ -ri -ri CO O 3 P -P d CD 00--101 Ό cd ft cd a) p P -P -ri> i cd cd md

3 U U CD Ίΰ P! K Mil S

01 • ri

• P 'CD

* CD Ë

• ri CD CD 'CD

P CD C P

'' CD P -ri U

+ J 3 -P 'CD

O -P (d

cd cd h + J <D

P P '(D d -ri V

cd ”cd Cn O cd cry U CP Ό rri o E cd -ri d CD“ ri Ë CD ce P Cd i — I rri

CD 'CD

P CD 'O rri CO E

CD CD CD d

CD OR d CD cd CD

E-C d Ο Ό Ό Ό CD Cti -ri rri CD -P CD CD d

rri CD ü CD d O

cd -ri Cti> 1 O -ri

> 0 ‘P rH -H -P

P P 'CD 0 4-1 cti

cd ο d pü E

+ J 01 Ό cd P

d CD -ri> i 'CD 0 H Ό PI K pi p4 ÇD.YD.MdC.8 - 9 - SY-1711 ΰ) T5 ”d) 4J MD m m” 54

.2 O -P

U 1H d) (D d> (D

□ 3 C 54 Ό Ό Ό

H D

E Cn d> +) 4J 4->

.. O d) · Η 1i-l 1H

t, 1 (D H H H3 · 1Ü · (O ·

; ^ + JH 54 (D 54 d> 54 (D

m IÖ-H 0) 4-> C 4-> G 4-1 G

ïï 54 3 Ό KO KO KO

i +10 d) 4-1 d) 4-1 d) 4-1 -h £ 4 ΐ -a - ft - & * f1 C 1H H S H J> 1 H>]

Cl 53 4-1 <ti 54 54 54 »t 'S C d) 54 110 4-1 '10 4-1 rtÜ 4-1

I S O -P Ό I I I

od: Η ^: κω <dd> d) di <D οι H ^ H d) d) O 03 54 0354 0354 t- -H ft- OO 3 03 03 ® 3 (Ö H 3 H> H> H> ON H 'd) d) MD O' (DO)

(D fflü'lDtJi ÜH 0 H UH

£ ü 3

Λ O

m 03 c

2 CO

Z ίΰ O '10 C

"rt H C

m d) (D

C. 0) (D 03 O -H

Ό 4-) -H C nJ

cm m E

.03 m ε 03 LÎ d) 4J 03

M 3 -, 03 1 -H O

D1 m 1H of O

• H 3 m p- 54 5D

n in S '<D U

p O ^ 54 MÖ Md ^ O '5 4-1 d) 0) 0)

M 05 C C ü 4-1 O

w 03 O 0) c C C

j> i -h E m m m H £ +1 '0) 03 · 4-1 dP 03 ·

m ft ü 54 03 dP (0H 03 O

w C MD -H O 1rl O 1H »

^ 03 Ό Ό O H 03 O O

^ (D '0) O 54 MD 1 54 m 3 p $ s u mû p $ ο o »m ^ O1

• H

4-> * 03 • rl 54 »0) 4J φ m cd> 1

54 d) N

m +> 0 um 03 54> 1

+1 HH

• r4 U

c mm

4-1 S5 H

03 03 d> <D »m wü E4 Ό d) d) c or O c c h mm 4J 4-1 4-1 O 03 (0

3 1H -H

»Ö 03 03

"(D MD" d) W

ci (¾ pî a ÇD.YD.MdC.8 -v 10 - SY-1711

TABLE III

Use of carbon sources by the H710-49 strain

Glycerol + D (-) Arabinose - L (+) Arabinose + "D-Xylose + D-Ribose + L-Khamnose + D-Glucose + D-Galactose D-Fructose + D-Mannose L (-) - Sorbose

Sucrose -

Lactose

Cellobiose + Melibiose

Trehalose +

Raffinose - D (+) - Melezitosis

Soluble starch +

Cellulose

Dulcitol -

Inositol - D-Mannitol + D-Sorbitol Salicin 1 CD.YD.MdC.8 - 11 - SY-1711

Observation after 3 weeks of incubation at 28 ° C. Basic medium: Pridham-Gottlieb inorganic medium.

The purified cell wall of strain H719-49 contains mesodiaminopimelic acid but no glycine. The whole cell hydrolyzate reveals the presence of madurose (3-0-methyl-D-galactose), glucose, ribose and a small amount of mannose. The composition of the cell wall and the sugars, which constitute the entire cell of the H710-49 strain show that the latter belongs to the type of cell wall IIIB.

The characteristics listed above of the H710-49 strain resemble those of the genus Actinoma-dura. In accordance with the digital taxonomy of Actino-madura and related actinomycetes established by Goodfellow et al. in Jj ^ Gen. Microbiol. 112; 95-111 (1979), most of the Actinomadura species existing in the soil fall into group 7 (Cluster 7) among the 14 which are described. The H710-49 strain is very related to the group 7 species. Nonomura and Ohara in Ferment. Technol. 49: 904-912 (1971) describe five saprophytic species of the genus Actinomadura and Nonomura £ ~ J. Ferment. Technol. 52: 71-77 (1974) _ ^ and

Preobrazhenskaya et al. ^ ÇActinomycetes and related Organisms 12: 30-38 (1977) _ ^ describe the identification; and the classification of Actinomadura species. According to the comparison with the known species of Actinomadura described in the literature, the strain H710-49 is considered to belong to a new species of Actinomadura similar to A ^ _ roseola, A. salmonea, A. vinacea or A ^ _ corallina , the description of which is given by Preobrazhenskaya et al. above and in Japanese document Kokai 55/9491.

It should be noted that for the production of BBM-1644, the present invention, although described in detail with reference to the particular strain H710-49 of Actinomadura sp. (ATCC 39144), is not limited to this CD.YD.MdC.8 - 12 - SY-1711 microorganism or to microorganisms described completely by the culture characteristics specified in this memo. The scope of the invention extends specifically to the H710-49 strain, as well as to its natural and artificial variants and mutants producing BBM-1644.

The antibiotic BBM-1644 in accordance with the invention "can be prepared by culturing a strain producing BBM-1644 belonging to the genus Actinomadura, preferably a strain d1Actinomadura sp. having the characteristics of identification of the strain ATCC 39144 or of a mutant thereof, in a conventional nutritive aqueous medium. The organism is cultured in a nutritive medium containing known sources of nutrients for actinomycetes, namely assimilable sources of carbon and nitrogen, in addition to possible inorganic salts and various other known growth factors. Aerobic aerosol culture is preferred for the production of large quantities of the antibiotic, but for the production of limited quantities, surface cultures and bottle cultures are suitable. The general techniques applied to the culture of other actinomycetes are applicable for the purposes of the present invention.

- The nutrient medium must contain an appropriate assimilable carbon source, for example glycerol, L (+) - arabinose, D-xylose, D-ribose, D-glucose, D-fructose, soluble starch, D-mannitol or cellobiose. As the nitrogen source, ammonium chloride, ammonium sulfate, urea, ammonium nitrate, sodium nitrate, etc. are suitable, both singly and in conjunction with organic nitrogen sources such as peptone, meat extract, yeast extract, maceration liquor- CD.YD.MdC.8 - 13 - SY-1711 corn, soy flour, cottonseed meal, etc. . The medium can also be supplemented, if necessary, with inorganic nutritive salts constituting sources of sodium, potassium, calcium, ammonium, phosphate, sulphate, chloride, bromide, carbonate, zinc, magnesium. , manganese, cobalt, iron, etc.

The production of the antibiotic BBM-1644 can be brought to any temperature ensuring satisfactory growth of the producing organism, for example 20 to 37 ° C, and is advantageously carried out at a temperature of about 27 to 32 eC. Usually, production is optimal after about 6 to 7 days of incubation in shaken flasks. For fermentation in tanks, it is advantageous to produce a vegetative inoculum in nutritive broth by inoculating the culture broth with a culture on agar or on earth or with a lyophilized culture of the organism. After having obtained in this way an active inoculum, it is aseptically transferred into the medium contained in the fermentation tank. The production of the antibiotic can be monitored by assay by diffusion on agar with paper disc in the presence of Bacillus subtilis M45 £ ”mutant Rec ~; Mutation Res. 16: 165-174 (1972) J as the testing agency.

At the end of fermentation, BBM-1644 is mainly found in the liquid fraction of the fermentation broth, after the solid fraction has been separated by filtration or centrifugation. The broth collected can therefore be separated by centrifugation into a mycelium cake and a supernatant broth. The filtrate is then concentrated and dialyzed against tap water using a semi-permeable membrane, for example a cellophane tube, to remove the permeable impurities. The solution retained CD.YD.MdC.8 - 14 - SY-1711 on the internal side (after elimination of the insolubles) containing the BBM-1644 can then be saturated with a salting-out agent such as ammonium sulfate which causes the BBM-1644 in the form of a crude solid.

The latter can be dissolved in water and then “desalted by dialysis against tap water.

The purification of the crude BBM-1644 can be continued according to the usual techniques for the other acid polypeptides. For example, the aqueous solution containing BBM-1644 can be adsorbed on an ion exchanger, such as DEAE-Sephadex, DEAE-

Cellulose, CM-Sephadex or CM-Cellulose, then be eluted with a neutral saline solution. It is preferable to carry out successive chromatographies with a concentration gradient of the saline solution serving as an eluent. The aqueous fractions containing the purified BBM-1644 are then concentrated to dryness, for example by lyophilization.

BBM-1644 is isolated by lyophilization in the form of an amorphous white powder. Reviewed by

electrophoresis on high voltage paper (4500 V

in 0.05 M barbital buffer at pH 8.6), BBM-1644 migrates as an acid at a rate of 8.7 cm per hour to the anode. BBM-1644 has no defined melting point and gradually decomposes above 240 ° C. It is soluble in water, but practically insoluble in common organic solvents such as methanol, ethanol, acetone, ethyl acetate and n-hexane. The antibiotic has a rotation

optical / o (72® from -75.6 ° in 0.25% aqueous solution. D

As shown in Fxg. 2, the UV spectrum of BBM-1644 1% has absorption maxima at 275 nm (Eicm 8.2) and 1% 310 nm (E ^ cm 4.6, shoulder) in aqueous solution. It has a practically identical UV spectrum in solution in water and in 0.01 N HCl, but only one CD.YD.MdC.8 - 15 - SY-1711 1% maximum at 285 nm (Eicm 8.9) in solution in 0.01 N NaOH. The IR spectrum of BBM-1644, recorded on KBr, constitutes FIG. 1. The spectrum reveals the presence of NH and OH radicals (3300 ^ 2980 cm “l) and amide radicals (1650 and 1540 cm” l). The antibiotic gives positive reactions of Folin-Lowry, to xantho-protein, to biuret and to ninhydrin and discolour the solution of potassium permanganate. It is negative to the reaction to the anthrone and to the Sahaguchi reaction. During cochromatography on Sephadex G-75 with ovalbumin (PM 43,000), chymotrypsinogen (PM 25,000) and ribonuclease A (13,700), BBM-1644 is eluted just after chymotrypsinogen and its molecular weight is as soon as then estimated at a value close to 22,000. Elemental analysis of BBM-1644 reveals 46.60% carbon, 6.45% hydrogen, 13.34% nitrogen and 0.20% sulfur. The determination of the amino acids in the BBM-1644 molecule reveals the presence of thirteen varieties of amino acids indicated in Table IV. BBM-1644 does not contain basic amino acids such as lysine, histidine and arginine.

TABLE IV

Amino acid composition of BBM-1644

Amino acid Relative composition; amino acids *

Alanine 8.8 - Aspartic acid 6.0 Hemicystin 1.0

Glutamic acid 5, 7

Glycine 8.7

Isoleucine 2.3

Leucine 1.0

Phenylalanine 1,4

Proline 4.1 Serine 1.6 CD.YD.MdC.8 - 16 - SY-1711

Threonine 7.2

Tyrosine 0.6

Valine 11il * The leucine content is arbitrarily chosen to be 1.0.

"

BBM-1644 is relatively stable in the pH range of about 2 to 9, but stability declines rapidly beyond this range. The aqueous solution of BBM-1644 is stable for 2 hours at 50 ° C at the pH of neutrality. Upon exposure to ultraviolet light, the antibiotic activity of BBM-1644 is lost within 20 minutes.

The physicochemical properties of BBM-1644 listed above indicate that it belongs to the anti-tumor protein antibiotics among which are neocarzinostatin, macromomycin and auromomycin. However, BBM-1644 can be differentiated from known protein anti-tumor antibiotics by its molecular weight, its amino acid content and its electrophoresis on paper. The electrophoretic mobility on paper of BBM-1644, neocarzinostatin and macromomycin are given "in Table V.

TABLE V

Electrophoresis on paper 1

Mobility 2 BBM-1644 +87 Neocarzinostatin +31

Macromomycin -20 CD.YD.MdC.8 - 17 - SY-1711 4. 500 V, 1 hour, barbital buffer of pH 8.6.

2

In mm from the initial spot.

Antibiotics from the neocarzino-statin group usually exhibit two UV absorption maxima at approximately 275 and 350 nm, while BBM-1644 exhibit UV maxima at approximately 275 and 310 nm. It was recently advanced in Biochem. Res. Common. 95: 1351-1356 (1980) that the maximum at 350 nm of the antibiotic neocarzinostatin could be due to the non-protein chromophores which are essential for their biological activity. It should be recalled that BBM-1644 includes a chromophore which is different from that of the known antibiotic neocarzinostatin.

The antibacterial activity of BBM-1644 is measurable by serial dilution of reason two in agar. Nutrient agar is used for gram-positive and gram-negative bacteria, nutrient agar with 4% glycerol for acid-resistant bacteria and Sabouraud agar for fungi. The activity is expressed by the minimum inhibitory concentration (MIC) in the agar and the results are presented in Table VI in comparison with those of neocarzinostatin. BBM-1644 exhibits potent inhibitory activity

S

against gram-positive and acid-resistant bacteria, but it does not inhibit the growth of gram-negative bacteria or fungi. The antibacterial spectrum of BBM-1644 is similar to that of neocarzinostatin, but the intrinsic activity of BBM-1644 outweighs that of neocarzinostatin for some of the test microorganisms.

CD.YD.MdC.8 - 18 - SY-1711

<D

c • H '

+ J

td ^ Φ CO H VD rH in

O’-fOfOO-HnOOOOOiNO

d R oooomrHO

^ * r-1 rH iH rH iH

g1 h Λ Λ A Λ Λ

ß (0 ^. U O

α 'Φ Φ s s

H

ü

O

R ^

d ^ ^ N (O IC li ffl LO rH

T VO ~ ^ *> -

? i-HOOVOrHrHOOOOOCHCOO

_, I O O O O H O

.5 S rH rH rH rH rH

> φ m Λ A A A Λ c d n 0)

ΰ -H

C Λ ** 0

w U

h 1 "£

<; S

(Ö

Eh 00 'R eu ij w. h a

d O O O

2 CH A OH rH CO

dm + J ο ο σ rH σο 7, 2: <-H CM O rH rH σ VO 00 Î ^ dQEriJrHCH | rij 00 a 0) 00 00 r ω h cm h vo id -h co "sj *

d mffiQ) UrHUbQ) com +) Q

βββΛΟ &, ΕΦ-ΦΟΦ 2 • φ φ φ φ h -Η σ β g · η <

MJH ^ raram! Aß <! - Hcr5a) H

Ό ß ß ο ß ß · η Ο 5ΐ ni Η m (T) (d> ((i)> H-HHtD3Eira g CU +) (d-HHß-HSHWWftß 5 CÜW ß rH +) Ο Φ Sh Φ (Ö “ßßißHiwcßUßidröEeo

’D U Ü ß £ ft tu 3 ß -H

5 ÜÜÜWCQßlÖ rHW-rl-H.Q

2, OOüßßin-HidßröMSHH

R1 dooüu, β η> β φ φ (d O OOUOUlOürH 0 + 1 +) ^ HrHOOO0-H0CQÉÜOfd

> <> H +, c; üHeH-rfßO (d (d'ß, β, β Cu O O rH φ ω φ Ό £ £ -H

& ΰ4Φ! π & Η · .Η £ ΑΑβΟΟ, ΰ ftiid} HÜUÜÜ0O0Uüß hj + j + i-HtH rötoH ^ co> i> ifd C0WC0SSeQlxli *; CUCU22U

CD.YD.MdC.8 r 19 - SY-1711 The ability of BBM-1644 to induce prophage in a lysogenic bacterium (ILB) is determinable by the technique of Lein et al. (Nature 196; 783-784, 1962) using neocarzinostatin as a control compound. The counting of the plates is carried out on agar containing the experimental compound (E) and the; control compound (T). An E / T ratio of plate counts greater than 3 is considered significant and the ILB activity is expressed by the minimum inducing concentration of the test compound. As shown in Table VII, the ILB activity of BBM-1644 is similar to that observed on neocarzinostatin, the minimum inducing concentration of which is 1 microgram per ml.

TABLE VII

Induction of a lysogenic bacterium by BBM-1644

ILB E / T activity * 100 10 JL 0.1 / ug / ml BBM-1644 10.4 10.3 6.0 1.2 Neocarzinostatin 28.6 20.4 10.8 0.7 * Significant E / T activity > 3.0.

The anti-tumor activity of BBM-1644 is determined in mice (strain BDFj) against P388 lymphocytic leukemia. Each mouse is inoculated intraperitoneally with 3 × 105 tumor cells. Phased doses of the antibiotic are administered to mice intraperitoneally 24 hours after implantation of the tumor. Treatments are carried out once a day on days 1, 4 and 7 (program q3dx3) or else 9 consecutive days (program qd 1- ^ · 9). Neocarzinostatin is administered as a control antitumor agent, the results being CD.YD.MdC.8 - 20 - SY-1711

V

collated in Table VIII. BBM-1644 is highly effective against mouse leukemia at doses ranging from approximately 0.03 to approximately 1.0 mg per kg per day depending on the two treatment programs. The anti-tumor activity of BBM-1644 is roughly the same (program qd 1—> 9) or 3 times greater (program q3dx3) than that of neocarzinostatin, to be expressed in minimum effective dose.

TABLE VIII

Anti-tumor activity against leukemia P388 E / T (%) in TSM *

Dose in mg / kg / day, ip, q3dx3 1_ CL_3 CL_1 0.03 0.01 BBM-1644 188 188 138 125 113 Neocarzinostatin 163 150 125 113 E / T (%) in TSM *

Dose in mg / kg / day, ip, qd 1- »9 0.3 0.1 0.03 0.01 BBM-1644 125 175 138 113 Neocarzinostatin 163 138 125 113 * Average survival time; significant activity E / T ^ 125.

The anti-tumor activity of BBM-1644 is also demonstrated by a second test against leukemia P388 in mice, the results of which are collated in Table IX. The details relating to the techniques applied for this test have been described in Cancer Chemother. Rep. 3i 1-87 (Part 3), 1972.

CD.YD.MdC.8 - 21 - SY-1711 fi w

I? VDVDVDV0 \ 0 \ 0 VDVDlDVDVDVDlOVO

rin N N N N \ V NNN \\ NNN

T yjVDV £> V £) V £) VO ^^ DVOVDVOVDVDVO

i s

“N 03 ci n m n n ri ci co ri co l · m S _ <k h k ^ kl ^ k h h Ik“ kl

^ -ί ci Μ H H | <J (N (N N

g I I I I I I I I I I I I I

+! „^ Σ \ cnj r · - o co ovo ^ fovDnr ^ M

ujSivT co γ-ι r- · vo in m omvocomoorHCN

00 η, n "ΗϊΗΉιΗγΗ rH» Hr—! F-HiHr ~! F — II — I

no ^ “m m * eu Φ H rn E oinmoino ootnmoomo U ïP? oommrorg

^ P k "1 e” l i — l iH r-l iH ιΗ ι — I “H fH r — I rH iH iH

H <D

i — I

_ "3

fi5 H

<h -rîs m IfltD'ÎN'CCO ^ 'M

D CT Φ CO N H O IDOJVOCOHOOO

[3 V) - ^ ............

OJ g1 H O O O O O CMrHOOOOOO

J <? in “J s) H e

THIS

s

B CQ

CQ <o r-l fi fl.

u g -H Γ "· t; ni P ^ ή, 2 ^ φ ·

S 0> ft H Q

S o rd

W U M Q

"Ai k (D

0) fi

H

4-> iti c s

î I

"• N ^

U VD

(Ö Ή

0 I

O 2

Ό) CQ

5 m CD.YD.MdC.8 -22- SY-1711 "i *

"I

C w

VDVDVOVDVDVDVDVDVDVOVDVDVOVDVOVD

VDVDVDVDVDVDVD \ OV0VDVDVDVOVDV £) V9

Sa 1 »S vd cninnoom <Tiinaic \ jc \ inc ^ infOcMr-i

'fn noi ^' nrocvjoj ^ -iin ^^ t '^ ronrMrH

ü ^ I I I I I I I I I I I I I I I I

P Eh

0) S \ HrHVOrHCnmrHHCTlrHHOCN '^ nrN

MHCflW VOrHOvÛrOnrHHOOHHOr ^ 'irOfNJ

IM Eh NMNHHHHH ΗΓΊΓΊΗΗγΗΗ

H <#P

® s “inoininoooo0oootnooo 2 n ooo ^ oo ^ cMCMOO ^ oc ^ oDirinoj ^ i

φ. "dj OïiHfHiHiHiHiHiH fHr-WrHrHiHi—! rH

X

H & v • H ^ T * OD ^ tN ^ CO ^ MHyfMVDCO'ÎCMHÎ! [3 *. £; CNVOCOiHOOOOVOCOHOOOOq ^ ra ^ hoooooooooooooOq S ^ w ° e

PI

CQ

(1) <! 3 a)

Eh g · Η Γ "σ>

E -P A

(0 Γ3 T

i M fl) ·· I-H

in & h

0 <ö D

MM O a (¾ »0) ϋ

Vf ü? ! *

"<CQ

CQ

CD.YD.MdC.8 - 23 - SY — 1711 _ φ “ο Η

cm S

ο ο ο ο g! Ρ Η iS rH rH ö Λ m \ \ \ \ § s Ο Ο Ο O 7d

H CN Η H .tJ

3 5 + J

w. ° g fÖ

V

'CD

00 -p S vo 1 O 1 1> * d CP · V Zi Ο Ό. 1 £ ü m -P E-i 2 im to W 1 1 1 1 5 H # ^ u

Φ P

Cn d - (Ö Ο Φ

Φ m Μ O 4J

• P £ OOOO -P Hit d?; co en h -Μ χ * Φ CD ^ ° Η H> - π CO -. * Φ

d Ln d P

Η “H

dl P Ο φ φ X w 3 d E E 'Φ H d? ΟΦ'ΦΕΗ • M 5 d · π> i -P O ft

* d d d d -H Ο O CO

-Ys co w co co des d> φ H *. it co ”φ φ CQC Φ dl B ln d

Co ^; ΡΦ-Μ ^ Μΰο · 'D p Η Η -Μ> »φ Ό · Η d H B t> d> p-P-HtP <l> CO Ω d H CD Ο ΪΜ

p itOdWfödHO

Φ PO OE

CQ -, Ό Φ Φ -P Ο -H

H CD T5 Φ CD CO

cdhh s ^>> h ό £ 2 ^ φ Φ P CO CO CN Ή, d · Η

Eh I Ti HEddEHH +) dO

löd C ^ voin ^ 5 ^ ra § ^ üd ^ feg, ° ° ° ° Η ® II ΛΙΙ-η ο Φ rHMrHr-l φ t0 LO IM -Η Ό

R £ ü -H \ Il E E -ri 4J

£, ¾ p ^ - ^^ ddd

»W Ϊ LOd S W W tn Η H

5 OOv / CQ ΗΟΠ3 + * HCO * E rt1 ä1 CO CO & i

|| I "|| "" ·· ·· · "II

P

d φ

E

d

-P

c φ Φ Ό d S, o 5 * E * φ -η ^ d +> + i φ d Η · Η (0 p • m d o d -p * φ

ο U φ · Η Η Φ -P

E O + JXföMH-H S

»Φ d <0 0>» p P d d

Eh H £ CEhWHU 10 Ü CD.YD.MdC.8 24 - SY-1711

The acute toxicity of BBM-1644 is determined in mice (strain dd Y) by single intraperitoneal administration, the LD50 calculated being 5.8 mg per kg.

As can be seen from the above indications, BBM-1644 exhibits potent antibacterial activity against gram-positive and acid-resistant * bacteria and is therefore useful for the treatment of mammals and other animals in which such bacteria have caused infections. In addition, it can be usefully administered in other traditional applications of antibacterial agents, such as the disinfection of medical or dental equipment.

The induction of prophage in lysogenic bacteria and the marked anti-tumor activity against mouse P388 leukemia reveal that BBM-1644 is also therapeutically useful for inhibiting the growth of tumors in mammals.

The subject of the invention is therefore a method of therapeutic treatment of a host animal suffering from a bacterial infection or a malignant tumor, according to which this host animal is administered, in an effective antibacterial or tumor inhibitory amount, " BBM-1644 or a pharmaceutical composition containing it.

According to another aspect, the subject of the invention is a pharmaceutical composition which comprises, in an effective antibacterial or tumor inhibitor amount, BBM-1644 together with an inert pharmaceutically acceptable excipient or diluent. These compositions can be presented in any pharmaceutical form suitable for parenteral administration.

The preparations according to the invention which CD.YD.MdC.8 - SY-1711 are intended for parenteral administration include in particular sterile aqueous or non-aqueous solutions, suspensions or emulsions. They can also be presented in the form of sterile solid compositions to be dissolved in water or sterile physiological saline solution or a. other sterile injectable medium immediately before administration.

It should be noted that the preferred amounts in fact of the antibiotic BBM-1644 vary with the nature of the composition, the mode and site of administration, the condition treated and the nature of the host. Many factors that modify the effect of the drug should be taken into account by the specialist, such as age, body weight, gender, diet, time of administration, route of administration, route of excretion, patient status, drug combinations, sensitivity reactions and severity of the condition. The administration can be carried out continuously or periodically within the limits of the maximum tolerated dose. The optimal therapeutic program in a given set of conditions can be evaluated by the specialist using standard dose-determination tests, taking into account the indications given above.

The invention is illustrated without being limited by the following examples. DEAE-Cellulose is an ion exchange diethylaminoethylcellulose. SEPHADEX G-50 is a filter gel produced by the company Pharmacia Fine Chemicals, Inc. DEAE SEPHADEX A-50 is a diethylaminoethylated anion exchange gel manufactured by the company Pharmacia Fine Chemicals, Inc. SEPHADEX® is a registered trademark of Pharmacia Fine Chemicals, Inc.

CD.YD.MdC.8 - 26 - SY-1711 EXAMPLE l.-

Preparation of BBM-1644 by fermentation.

A culture on an inclined agar bearing a well-established colony of Actinomadura sp. H710-49 to inoculate a seeding medium (100 ml * in an Erlenmeyer flask) containing 1% mannitol, 2% peptone and 1% yeast extract, the pH having been adjusted to 7.2 before sterilization. The seed culture is incubated at 32 ° C for 72 hours on a rotary shaker (250 rpm) and 5 ml of the culture are transferred to the second seed medium (100 ml) having the same composition as the first. The seeding culture is carried out under the same conditions as the first. 5 ml of the resulting inoculum is used to initiate fermentation in 500 ml Erlenmeyer flasks containing 100 ml of fermentation medium containing 2.5% mannitol, 0.5% glucose, 1% soy flour, 0, 5% peptone, 1% meat extract, 0.3% CaCO3 and 0.2% NaCl. Fermentation is carried out at 28 ° C on a rotary shaker at 250 rpm. The production of the antibiotic is monitored by assay by diffusion on agar with paper disc in the presence of Bacillus. lus subtilis M45 (mutant Rec ”) as a test organism. The antibiotic activity in the culture broth gradually increases with the advancement of the fermentation and reaches approximately 300 μg per ml after 6 or 7 days.

EXAMPLE 2.-

The collected broth (18 liters) obtained by fermentation in Example 1 is separated into a mycelium cake and a supernatant broth using a Sharpless type centrifuge (Kokusan No. 4A). The filtrate is concentrated below 40 ° C to one tenth of the initial volume and the concentrate is dialyzed in a tube CD.YD.MdC.8 - 27 - SY-1711 λ of Cellophane (Union Carbide) against water of distribution in cold room. The solution retained on the inner side is concentrated to approximately 1.5 liters and centrifuged (at 8000 G) to separate the insolubles. The clear supernatant * is saturated with ammonium sulfate and allowed to stand for 5 hours at 5 ° C. The resulting precipitate is collected by centrifugation which is dissolved in water (300 ml) and desalted by dialysis against tap water. The dialyzed solution (700 ml) containing 22 g of crude solid BBM-1644, as indicated by the lyophilization of an aliquot of the solution. The rest of the solution is used for purification without concentrating it to avoid decomposition. The solution of the antibiotic is passed through a column of DEAE-Cellulose (Cl-, 400 ml) and the solution is washed with water (1 liter), then it is developed with phosphate buffer 1/15 M (pH 7.0) containing 0.3 M sodium chloride. The active fractions (300 ml) are combined and dialyzed for 18 hours against tap water, then they are chromatographed on a DEAE- column. Cellulose (400 ml) which has been balanced with 1/15 M phosphate buffer (pH 7.5). The column is developed with the same buffer containing increasing amounts of sodium chloride (0 to 0.2 M). The eluate is desalted by dialysis and deposited on a column of DEAE-Sephadex A-50 (17 ml). The column is developed with 1/15 M phosphate buffer (pH 7.5) by establishing a sodium chloride concentration gradient (0 to 0.3 M). The fractions appearing active against B ^ subtilis M45 are pooled and dialyzed against tap water for 18 hours. The desalinated solution is chromatographed on a column of DEAE-Sephadex A-50 (18 ml) using a buffer system with CD.YD.MdC.8 - 28 - SY-1711 1/15 M phosphate (pH 7.0) - NaCl (0 to 0.3 M) as eluent. The appropriate fractions are collected, concentrated to 10 ml and the product is placed on a column of Sephadex G-50 to desalt it. The column is eluted with deionized water and the active eluate is lyophilized to collect 120 mg of a white powder.

* The BBM-1644 sample thus obtained is homogeneous, as indicated by the polyacryl-amide gel electrophoresis.

* »CD.YD.MdC.8 - 29 - SY-1711

Claims (9)

1, - The antibiotic BBM-1644 which (a) is effective in inhibiting the growth of grain-positive and acid-resistant bacteria, (b) is effective in inhibiting the growth of P388 leukemia in mice, (c) induces the prophage character of lysogenic bacteria, (d) is soluble in water but practically insoluble in methanol, ethanol, acetone, ethyl acetate and n-hexane, * (e) present an infrared absorption spectrum (KBr) substantially as illustrated in FIG. 1, (f) has an ultraviolet absorption spectrum in water, 0.01 N HCl and 0.01 N NaOH substantially as illustrated in FIG. 2, (g) has an optical rotation of -75.6 ° in 0.25% aqueous solution, (h) has no defined melting point, but decomposes gradually beyond 240 ° C, ( i) moves approximately 8.7 cm towards the anode during electrophoresis on paper at 4500 V for 1 hour in 0.05 M barbital buffer of pH 8.6, ”(j) presents the analysis following elementary: a C, 46.60%, H, 6.45%, N, 13.34%, S, 0.20% and O (by difference) 33.41%, (k) is a high peptide molecular weight, the molecular weight of which is estimated at approximately 22,000, (l) discolour the potassium permanganate solution and give positive reactions from Folin-Lowry, to xanthoprotéine, to biuret and to ninhy-drine and negative reactions to 1 anthrone and Sakaguchi, and (m) reveals by hydrolysis the following composition CD.YD.MdC.8 - 30 - SY-1711 in amino acids, based on a leucine content arbitrarily fixed at 1.0: alanine ( 8.8), aspartic acid (6.0), hemicystine (1.0), glutamic acid (5.7), glycine (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). % 2. Method for producing the antibiotic BBM-1644, characterized in that a strain of Actinomadura sp. producer of BBM-1644 in an aqueous nutritive medium containing assimilable sources of carbon and nitrogen in aerobic submersion until production of a significant quantity of BBM-1644 by this organism in this culture medium. 3. Method according to claim 2, characterized in that the BBM-1644 is isolated from the culture medium. 4. Method according to claim 2 or 3, characterized in that the organism producing BBM-1644 has the identifying characteristics of Actino-madura sp. H710-49 (ATCC 39144). 5. Biologically pure culture of the microorganism of Actinomadura sp. ATCC 39144, this culture being capable of producing the antibiotic BBM-1644 in an amount which can be isolated by culture in an aqueous nutritive medium containing assimilable sources of nitrogen and carbon. 6. A method of therapeutic treatment of a host animal suffering from a bacterial infection, characterized in that BBM-1644 is administered to this host animal in an effective antibacterial dose. 7. A method of therapeutic treatment of a host animal suffering from a malignant tumor sensitive to BBM-1644, characterized in that this host animal is administered BBM-1644 in a dose inhibiting the tumor. CD.YD.MdC.8 - 31 - SY-1711 * 4 8. Pharmaceutical composition, characterized in that it comprises BBM-1644 in an effective antibacterial amount, together with a pharmaceutical excipient or diluent. 9. Pharmaceutical composition, characterized in * that it comprises BBM-1644 in an effective amount * inhibiting tumors, together with an excipient% or pharmaceutical diluent. kr «H c CD.YD.MdC.8 - 32 - SY-1711