Note: Descriptions are shown in the official language in which they were submitted.
~ ~ - 1070675
The present inventlon relates to-salts of polymyxine
methyl tetracyclines having the characteristic formula:
- .... : .- ,, ~ .' - : . ,
[(TH - C0 - NH - CH2 - NH)n - P, nA ], mHB (A)
in which~
; m and n represent positive integers less than 6, -
A and B represent mineral or organic anions,~
`T represents a tetracyclical antibiotic residue of the tetra-
cycline family having a prlmary amide function, the formula of
- which is T - C0 - NH2, such as tetracycline, oxytetracycline,
dimethylchlorotetracycline, methacycline, rolitetracycline,
doxycycline, minocycline,
P represents the residue of a colistine molecule, the formula
of whlch is P - (~H2)n.
The tritetracycline methyl polymyxine trichloro-
; hydrates have previously been described, but they are not usable
industrially because of their instabillty both~in dry-condition
and in solution.
The salts of this invention are remarkable in that
they are stable in dry condition and even in neutral or slightly
alkaline solution, for example at pH 8.5.
The present invention also relates to products respond-
ing to the above formula (A) to trioxytetracycline methyl coli-
stine trichlorohydrate, hereinafter called Negafongine.
~ The present invention also relates to the methods for
the preparation of new salts of the invention, characterized by
initially reacting formol and an antibiotic of the tetracycline
family in the form of its salt, and then reacting the product
with colistine sulfate.
-~ According to the invention, a reaction medium is sel-
ected both for the initial addition and for the final
~$
1070675
condensation which does not react with formol, and although
this temperature is not characteristic of the invention, gen-
erally the opexation is carried out between room temperature
and 60C. ~n fact, above this temperature, the antibiotic
substrata co~mence to degrade.
The present invention moreover relates to medica-
tions usable in human or veterinary medicine, characterized
by containing, as primary or secondary active principal, one
or more salts of the invention of the above formula (A). In
;~ 10 fact, at variable doses, these compounds are bacteristatic,
;~ bacteriocides on the Gram+ and Gram- bacteria, and on the
large viruses, the protozaires and certain flagellates.
The interest in the salts of formula (A) of the in-
vention resides particularly in the fact that the toxicity of
the polymyxines is considerably reduced in this type of com-
pound and that the anti-bacterial activity of the new com-
pounds of the invention is higher, in vitro and in vovo, than
the simple mixture of antibiotics from substrates for the syn-
thesis of the new salts of the invention. In vivo, in particu-
~0 lar, it is surprising to note that the new salts of the in-
vention completely pass the gastro-intestinal barrier, thus
pass into the interior medium and thus make it possible to
treat Gram- a~fections outside the digestive tract, such as
urinary affections, while it is well known that tetrac~clines,
which pass the gastro-intestinal barrier, although poorly,
are inactive and that the polymyxines do not pass this barrier
and thus cannot act outside the digestive tract when they are
administered orally.
The present invention relates in particular to med-
3 ications for human or veterinaxy medicine, characterized by
. : ~
:; ~
1070675
the fact that their activity is antibiotic and that they con-
tain at least one salt of this invention of fo~ula (A) above
as primary or secondary active principal.
Moreover, the present invention relates to the phar-
maceutical compounds containing at least one salt of the in-
vention of above formula (A).
In addition, the present invention relates to the
pharmaceutical forms, such as wafers, powders, gellules,
compresses, lozenges, syrup for oral administration, to the
form for administration in ophthalmology, otorhinolaryngology,
such as drops, pommades, cones, to the forms of rectal admin-
istrations, suppositories, enemas, to ready to use injectable
forms or those to be prepared at the time of use.
i The invention will be better understood from the
following example ~iven as a non-limitative case and describ-
ing the preparation and properties of one of the sales of the
invention, defined above under the name of Negafongine.
PREPARATION OF NEGAFONGINE
56 grams oxytetracycline chlorohydrate is put into
suspension in 6 liters of pure anhydrous alcohol. Heat to
50C and introduce, under agitation, 20 cc formol, previously
concentrated by one third and filtered to eliminate the insol-
ubles. The dissolution is complete in 15-20 minutes. Then ~ -
evaporate to dryness under vacuum.
The crystals obtained are taken up with 50 cc abso-
lute methanol after expansion in the air for a few hours to
eliminate the formol odor. Add directly to this solution,
50 grams colistine sulfate to avoid the formation of lumps.
Heat to 45-50C under agitation. The dissolution is complete
in 10~15 minutes. Some insoluble flakes are filtered and the
~070675
filtrate is left to stand overnight at -10C. The Nega0ngin2
obtaine~ is dried cold. 15-20% of product is still found in
the mother liquors.
The Negafongine is water-soluble in stable form but
has no clearly determined melting point because it decomposes
when hot.
Spectroscopic-Analyses, performed essentially in
comparison with those of the substrate having been used dur-
ing the synthesis, that is to say oxytetracycline chloro-
hydrate and colistine sulfate.
Infrared Analysis by dispersion in Nujol.
Considerable differences exist between Negafongine
and oxytetracycline chlorohydrate in the area of 3500-3000
cm-l. Particularly the tetracyclinical band at 3400 cm~l no
longer appears in Negafongine, which proves that the -NH2 group
of the oxytetracycline amide function has reacted, which is
still confirmed, although this area is charged by differences
observed in the region of 17U0-1600 cm~l.
Analysis by Nuclear Magnetic Resonance: by dissolu-
tion in deuterized dimethyl sulfoxide.
The spectrum of Negafongine has a mass in the strong
fields which is absent in each of the starting substrata. It
corresponds to the alcoyl radical hydrogens, proving the real-
ity of the new Negafongine structure. In the region of the
aromatic process, Negafongine maintains unchanged the oxy-
tetracycline phenolhydroxyl, which implies that the addition
of formol was not made in this area.
TOXICITY OF NEGAFONGINE
DL 50 per os male mice: 1900 mg/kg
DL 50 intravenously male mice: 22.5 mg/kg
aa/æmar4
1070675
DL 50 intramuscular male mice: 125 mg/kg
These toxicological studies w~re completed by worX
on different galenic forms: "dry-fill" by intramuscular in-
jection, "dry-fill" by intravenous injection, ready to use
injectable solution, gelatin-coated pill for oral use, going
from acute toxicity to chronic toxicity in mice, rats, rab-
:
bits and dogs. At doses 2.5 times higher than those provid-
ed in man, the tolerance always has been perfect, but at doses
still 10 times higher, that is 25 times higher than those pro- -
;~ 10 vided in man, a high mortality rate is observed (40/0 in rats
i 63 days after treatment), a relatively high reduction of the
number of red blood corpuscles and rather severe gastric
seizures also were noted.
~ ACTIVE ANTI-MICROBIAN ACTIVITY IN VITRO OF NEGAFONGINE
'd` ~ 15 On Gram+ and Gram- bacteria of hospital origin,
tests were conducted according to the known technique and the
disc (gelatin-coated medium). The activities of the colistine
and of oxyte~racycline are found with a synergizing activity
which manifests itself particularly on the Aeruginose ~Gram-)
'~ 20 P.S.
Results of the Clinical Tests
The first clinical tests regarding Negafongine were
-cQnducted by oral administration (gelatin-coated pills at 250
mg), ~y intramuscular ready to use injection or "dry-fill",
by intravenous injection. These results are perfectly satis-
factory, particularly as far as oral administration is con-
cerned, where it is known that the colistine cannot pass the
gastro-intestinal barrier. In fact, negafongine proved to be
active orally on Gram- affection of the respiratory, the
3 urinary and the gynecological tracts, and of course the di-
gestive tract.
-5-
