Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
1 164~64
This application is the seoond of three divisional
ap~lications of copending application serial no. 362,485,
filed October 16, 19800
BACKGROUND OF THE INVENTION
.
The characteristics and preparation of the anti-
biotic lincomycin are disclosed in U.S. Patent 3,086,912.
Clindamycin is disclosed in U.S. Patent 3,496,163. These
antibiotics have been extensively used as medicines in
humans and animals. A number of patents world-wide have
issued concerning these antibiotics and a variety of
derivatives thereof.
Lincomycin has the following structural formula
CH3
¦ ÇH3
HO-~-H
~ C N - ~
~ S~H,
H OH
Clindamycin has the following structural formula
cb/
1 1~4864
CHJ
H H_cH
N ~ ~
H~SCH3
H OH
BRIEF SUMMARY OF THE INVENTION
This application relates to noyel
and useful compounds of the general formula:
2 ~ Rl
E \ / VI
B
wherein ~, B and E are nitrogen, CH3-N=, oxygen, sulfur or CRlRl;
wherein Rl, which can be singly or multiply attached to
any ring carbon atom not already substituted by R2, is
hydrogen, alkyl and substituted alkyl, wherein the alkyl
portion is from 1 to 8 carbon atoms, and isomeric forms
thereof, cycloalkyl and substituted cycloalkyl, substituted
- 2 -
ma~/
1 164864
oxygen, substituted nitrogen, halogen, phenyl and sub-
d henyl or -(CH2) -OH, -(CH2)m NR4 5,
forms thereof, wherein m is an integer of from 1 to 8,
and R4 and R5 are hydrogen or alkyl of from 1 to 8 carbon
atoms and isomeric forms thereof; and wherein R2, which
can be attached to any ring carbon, not already substituted
by Rl, or nitrogen atom, is:
R
--c--x
and X is the amino function of 7(R)-hydroxy-methyl l-thio-
~-lincosaminide, 7(S)-hydroxy-methyl l-thio-~-lincosaminide,
7(S)-halo-methyl l-thio-~-lincosaminide, 7(~)-halo-methyl
l-thio-~-lincosaminide, 7(S)-methoxy-methyl l-thio-~-
lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl l-thio-~-
lincosamini~e, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl
l-thio-~-lincosaminide or 7-deoxy-7(S)-(3-hydroxypropylthio)-
methyl 1-thio-~-lincosaminide.
This application also relates to novel and useful
compounds, which are claimed in the above noted parent
application, of the general formula:
~(CH2)n~ 1
~R2
7 II
R3
wherein Rl, which can be singly or multiply substituted in
the 2, 3, 4, 5, 6, 7, 8 or 9 ~sition of the ring not already
substituted by R2, is as defined above; wherein R2, which
-- 3 --
mab/
1 16486~
can be singly substituted in any position of the ring not
already substituted by Rl, is as defined above; wherein R3
is hydrogen, methyl, ethyl or C2H5OH; and wherein n is an
integer of from 1 to 4.
This application further relates to novel and use-
ful c ~ ounds, which are claimed in a first divisional application,
serial no. 414,643, filed Nove~iber 1, 1982, of the general fonmlla:
~
~ N ~
wherein Rl, which can be singly or multiply substituted in
any position of the pyridine ring not already substituted
by R2, is as defined above; and wherein R2, which can be
singly substituted in any position of the pyridine ring not
already substituted by Rl, is as defined above
- This application still further relates to novel
20 and useful c ~ ounds, which are claimed in a third diyisional a~plication,
serial no. 414,645, filed Noveniber 1, 1~82, of the general fornmla:
~Rl
R2~
~ VII
B
- 4 -
i
mab/ !,
1 16486~
wherein A, B, D and E are nitrogen, oxygen, sulfur or
CRlRl; wherein Rl, which can be singly or multiply attached
to any ring carbon atom not already substituted by R2, is
as defined above; and wherein R2, which can be attached to
any ring carbon, not already substituted by Rl, or nitrogen
atom, is as defined above.
Compounds of particular importance are of the
general formula:
R1
~ H2)n ~ R
J III
N
R3
wherein Rl is in the 4 position and is alkyl of from 1 to
8 carbon atoms and isomeric forms thereof; wherein R2 is in
the 2 or 3 position and is otherwise as defined above; and
wherein R3 is as defined above.
Important precursor compounds of the above have the
general foxmula:
R
~ N ~ IV
wherein Rl and R2 are as defined immediately above.
The pharmaceutically acceptable acid ad~ition salts of the
a~ove no~d com~ounds including the 2-~hos~hates and 2-almitates,
-- 5 --
mab/~l~
1 16486~
wherein the substituent. is attached to thc oxygen atom at
the 2 position of the sugar ring of the above compounds, are
also described and claimed in this and the above noted
related applications.
The synthesis of the novel analogs described
herein can be shown in exemplary form as follows:
HC-Cl ~ 0 HC-HCZ
C-OH ~ ~H ~ CH ~ H
H0 ~ ~ H0 ~ ~
~ S-CH3 ~ S-CH3
CzHs k~
CH3
0 HÇ-Cl
N~ ~C-N~-CH
\l
~ H ~S-CH3
nH
The wavy lines denote either the D-cis or L-cis isomer.
An alternate procedure which may be used to syn-
thesize the novel analogs described herein can be shown in
exemplary form as follows:
C2Hs CzHs
/ + 7-Cl-MTL~ -7-Cl-MTL
H H
mab/ !:~
1 ~6~864
The wavy line denotes either the D-cis, L-cis, D-trans or
L-trans structures.
The L-cis structure
C2H5
H3
~N lc --N H--CH ~ CH l O
H~lk 0~1
~C1~3 y
OH
has been shown to be 5 to 10 times more active than clinda-
mycin against S. aureus and S. hemolyticus in laboratory
mice.
An isomer of V may be isolated from the above
reaction and is presumed to be the D-cis compound, VA.
The D-cis structure i`s not as potent an antibacterial agent
as the L-cis compound.
C=2H5
CH3
J JyO H~C-Cl
~-CH3 V A
.. .. .. . . . ..
DETAILED DESCRIPTION OF THE INVENTION
Upon reacting an amino acid of the formula
- 6a -
~ab/ `l`
1~6486 ~
7 3809A
C-OH
s
N
wherein R1, which can be singly or multiply substituted-in any
position of the pyridine ring not already substituted by
-C-OH ,
is selected from the group consisting of hydrogen, alkyl and substi-
! tuted alkyl wherein the alkyl portion is from 1 to 8 carbon atoms,
inclusive, and isomeric forms thereof, cycloalkyl and substituted
cycloalkyl, substituted oxygen t SU bstituted nitrogen, halogen, phenyl
and substituted phenyl; -(CH2)m-OH, -(CH2)m-NR4R5, and isomeric
forms thereof, wherein m is an integer of from 1 to 8, inclusive,
R4 and R5 are H or alkyl of from 1 to 8 carbon atoms, inclusive,
and isomeric forms thereof, wherein
O
-C-OH
which can be singly substituted in any position of the pyridine ring
not already substituted by R1, with a sugar amine compound selected
from the group consisting of 7(R)-hydroxy-methyl l-thio-a-lincOs-
aminide, 7(S)-hydroxy-methyl l-thio-~-lincosaminide, 7(S)-halo-
methyl l-thiO-a-lincosa'minide, 7(R)-halo-methyl l-thio-a-lincos-
aminide, 7(S)-methoxy-methyl l-thiO-a-lincosaminide, 7-deoxy-7(S)-
(methylthio)-methyl l-thio-a-lincosaminide~ 7-deoxy-7(S)-(2-hydroxy-
ethylthio)-methyl l-thio-~-lincosaminide, and 7-deoxy-7(S)-(3-
hydroxypropylthio)-methyl l-thio-a-lincosaminide; there are obtained
novel and useful compounds of formula I.
Upon reacting an amino acid of the formula
(CH2) ~ d
~ ~ C-OH
~ N
R3
1 16~864
-8- . 3809A
wherein R1 and the position of substitution of -C-OH are as defined
above; wherein R3 is selected from the group consisting of H, CH3,
C2Hs, and -CH2-CH2-OH; wherein n is an integer of from l to 4,
inclusive, with a sugar amine compound, as defined above, there are
obtained novel and useful compounds of formula II.
Upon reacting an acid of the formulae
HO-C ~ R1 `
E A
\ B
// ~ D
B
wherein A, B, D and ~ are selected from the group consisting of
nitrogen, oxygen, sulfur and CR1R1; R1 is as defined previously
and can be attached to any ring carbon or nitrog~n atom; R1 can
be multiply attached to any ring carbon atom; -Cr-OH can be attached
to any ring carbon or nitrogen atom, with a sugar amine compound
selected from the group as defined above, there are obtained novel
and useful compounds of formulae VI and VII.
MTL is methyl l-thlo-a-lincosaminide of the formula
CH3
HO-C-H
HzN - CH
H ~ ~
CH3
OH
epi-MTL is methyl 7(S)-7-deoxy-7-hydroxy-l-thio-~-lincos-
aminide of the formula
1 16~86~
-9- 3809A
CH3
H-C-OH
H2N CH
\~SCH3
OH
7-Cl-MTL is methyl 7(s)-7-deoxy-7-chloro-l-thio-a
lincosaminide of the formula
f
1 5 H-C-Cl
H2N - - fH
HO~O \j
\~CH3
OH
epi-7-Cl-MTL is methyl 7(R)-7-deoxy-7-chloro-1-thio-
~-1 incosaminide of the formula
ICH3
Cl -C-H
H2N ----- CH
~
H ~1 \1
- ~CH3
OH
7~S3-methoxyr~ethyl 1 -t~o-c.-l ~ncvsamtn;~de c~n be
shown as follows (See U.S. 3,702,322, Example 1, Part B-l):
~ ~6486~
-1 O- 3809A
IH3
H-C-O-CH3
NH2-CH
H ~ ~
S-CH 3
OH
With reference to the following formula, 7-deoxy-7(S)-
(methylthio)-methyl l-thio--lincosaminide exists when R
is CH3; 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl l-thio-
-lincosaminide exists when R is -CH2-CH2-OH; and, 7-
deoxy-7(S)-(3-hydroxypropylthio)-methyl l-thio--lincos-
aminide exists when R is -CH2-CH2-CH2-OH (See ~.S. 3,915,954, Examples
1, 10 and 31):
CIH3
H-C--S-R
NHz-CH
Hp ~ ~
S-CH3
OH.
The hydroxy and halo groups at the 7 position of
the above formulas can be shown as follows
CH3
~I~Y
HzN ~ CH
H ~ 0
~ CH3
OH
1 16~86~
~ 3809A
wherein Y is selected from the qroup consisting of 7(R)-
hydroxy, 7(S)-hydroxy, 7(S)- halo , and 7(R)- halo
When a pyridine acyl group is used, the resulting
analog can be reduced to give a mixture of the corres-
ponding saturated compounds, one of which is the L-cis
isomer. Other compounds which may be present include
the L-trans, D-cis, and D-trans isomers. Generally,
for any of the compounds descri~ed herein, the reduced
form is mare antibacterially-active than the unsaturated
precursor. The use of a piperidine acyl group gives
analogs existing as D-cis, L-cis, D-trans, and L-trans
isomers. Again, the L-cis isomer has been found-to be
~ore anti-bacterially active.
The general method used herein to prepare the novel
analogs is the well known process wherein an appropriate
acid is coupled with an appropriate sugar amine.
~'~xed Carboxylic Acid Anhydride Procedure," Chemistry of
The Amino Acids, Vol. 2, p. 970, John Wiley and Sons, Inc,
1961.) When the acid is unsaturated, the resulting
unsaturated analog can be catalytically reduced under
standard conditions to prepare the saturated analog. For
example, the reduction can be conducted using the follow-
ing conditions:
H2 at 5 to 50 psi
~5 Catalyst - platinium oxide (PtO2)
Solvent - H20 or H20 + MeOH, or H20 + EtOH
HCl - 10~ excess
Time - 24 to 48 hours
As used ~e~ein, alk~l of 1 to 8 carbon ato.~s~?
inclusive, and isomeric forms thereof, includes methyl,
ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and
branched chain isomers thereof.
Substituted alkyl means the above alkyl compounds in
which one or more of the hydrogen atoms has been replaced
by a halogen, i.e., Cl, Br, F, and I, oxygen, hydroxyl,
amine (primary), amine (secondary-alkyl substituted by
alkyl as above), amine (tertiary-alkyl substituted by
1 16~86~
-12- 3809A
alkyl as above), sulfur, -SH, and phenyl. Exemplar~
compounds are l-fluoroethyl, l-chloroethyl, 2-fluoroethyl,
2-chloroethyl, l-bromopropyl, 2-iodopropyl, l-chlorobutyl,
4-fluorobutyl, and 4-chlorobutyl.
Cycloalkyl ~eans cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl and cyclooctyl.
Substituted cycloalkyl means a cycloalkyl substituted as above
for substituted alkyl. Exemplary compounds are 2-cyclopropylethyl,
3-cyclobutylpropyl, 4-cyclopentylbutyl, and 4-cyclohexylbutyl.
Aromatic means phenyl and substituted phenyl wherein
one or more of the hydrogen atoms has been replaced by a
halogen, as above, hydroxyl, amine (primary, secondary,
and tertiary with the latter two alkyl substituted as
above), -SH, and phenyl. Exemplary compounds are p-
bromophenyl, m-iodophenyl, o-chlorophenyl, p-ethylphenyl,
m-propylphenyl, o-methylphenyl, and p-octylphenyl.
As detailed infra, the compounds of the invention
can be phosphorylated to 9ive the 2-phosphate, and
acylated to give the 2-palmitate which are both anti-
bacterially-active ln vivo.
Substituted oxygen means oxygen substituted by an alkyl of
from 1 to 8 carbons, inclusive, aryl, and substituted aryl.
Substituted nitrogen means nitrogen substituted by an acyl of
from 2 to 18 carbons, a monoalkyl of 1-8 carbons, inclusive, and a
dialkyl, wherein the alkyl is from 1 to 8 carbons, inclusive, includ-
ing the isomeric forms for all acyl and alkyl groups.
Halo means chloro, bromo, iodo, or fluoro.
Exe~plary sources for the amino acids used as starting
materials herein are as follows:
1. Heterocyclic Compounds, Vol. 1, John Wiley and Sons, Inc.,
1950. This source describes the preparation of halogen
and alkyl substituted amino acids.
2. Chem. Abstracts:
81 - 105223A - alkyl and cycloalkyl
81 - 152243S - alkyl and halogen substituted
8Z - 170746H - halogen substituted
85 - 46322Q dihalo substituted
1 16486~
13 3809A
85 - 177258W - dihalo substituted
84 - 116928X - dihalo substituted
81 - 3737d - phenyl substituted
78 - 58201t - phenyl substituted
76 - 126800y - tetrahalo substituted
82 - 11036K - bromo substituted
_ - 27119W - bromo substituted
84 - 16613X - bromo substituted
78 - 123494G - bromo s~lbstBtuted
CH2CH2=CH-CH3
84 - 135488Y - I II
~N "-`COOH
CH3-O
81 - 151951J - ~ lI
~ Br~`~ ~ COOH
Br-CH
81 - i7809a - I ~
N COOH
O-CH3
84 - 30918G - ~
COOH
C2Hs-O ~ o-C2Hs
81 - 33139C - l ll
..~N'~ COOH
I
79 - l9109Y -
. ~`N ~ ~COOH
1 16486~
-14- 3809A
81 - 37370
~N ~ COOH
81 - 135964K - ~ ~ COOH
NH-C-CH3
85 - 177349B - ~ OOH
OC2Hs
~ COOH
78 - 71865G -
Cl ~ ~N ~
81 - 135964K - Cl ~ o ~ COOH
COOH
83 - 147397G - \
-- ~N Cl
CH3
CH3
82 ~ 11036K - ~ NH2
~ COOH
1 16486~
-1 5- 3809A
84 - 11 6928X - Cl~COOh
C2Hs-O~,~ Q-C2H5
81 - 331 39C - ~N~OOH
Cl
7 6 - 1 26800Y - ~ Cl
Cl N COOH
p-C2H5
C51~
79 - 11 5449b -
N COOH
I~IH-CH3
Cl~l
67 - 63229K
N COOH
CH3
68 - 1 04926b ~3~
CH3 N COOH
6 9 - 5 9 04 8 Z ~S~
COOH
1 16486~
-16- 3809A
71 - 124907M H-~ C-OH
OH
68 - 59465N 1~ 0
O~N C-O-CH3
H
This compound can be hydrolyzed to the acid
by means well known in the art, which acid
can then be reduced, also by means well
known in the art.
86 - 106501 e l~<CH3
C C_N
H3
This compound can be hydrolyzed to the acid
by means well known in the art. The resul t-
ing acid then can be N-demethylated by the
procedures disclosed in U.S. Patent 3,583,972.
69 - 67282M CH3-l~ 1l
~N C-O-C2Hs
CH3
1 164864
-17- ` 3809A
This compound can be hydrolyzed to the acid
by means well known in the art.
CH3
90 - 7029/-14-Z CH3~
CH3
This compound can be hydrolyzed to the acid
by means well known in the art. Also, one
or both of the N-CH3 groups can be removed
from the resulting acid by fo110wing the
procedures disclosed in U.S. Patent
3,583,972.
~ ~ ~
90 - 168488X O~Nl C-O-C4Hg
CH3-f-H
0
This compound can be hydrolyzed to the acid
by means well known in the art. The result-
ing acid can be converted to the following
compound
~ O
~N ~ C-OH
H
by methods disclosed in U.S. Patent
3,583,972.
/=\
85 ~ 142995G NH2 ~ CH2 ~
~ COOH
1 164864
~ 3809
81 - 15202~S CH3-CH-CH2-CH2 ~
~ COOH
75 - 110156M ~
CH3-fH-CH2 N COOH
OH
Compounds having free NH2 or OH groups will have to
have these groups protected before being condensed with
the amino sugar. Protection of such groups is well known
in the art. See Protective Groups in Organic Chemistry,
J. F. W. McOmie, Plenum Publishing Co., Ltd., 1973.
3. Jour. Chem. Soc. 1969 - 2134 -
Various H-alkyl substituted pyridines
C-N ~-R CH2R
~ + R-MgX
(commercially
available)
R - alkyl, branched alkyl and cycloalkyl
4. Jour. Chem. Soc. 1969 - 934 -
R R R R R
NH~ ~ r ~ CsN ~ C-OH
1 164864
-19- 3809
The following examples are illustrative of the
process and products of the invention, but are not to be
construed as limiting. A11 percentages are by weight and
all solvent mixture proportions are by volume unless
otherwise noted.
Example 1 - 4-Cis-ethyl-~-pipecolic acid amide of 7-Cl-
MTL HCl (U-57,930E - Compound V~:
PART I
C2Hs HCl C2H5CH3
~ O ~ 7-C1-MTL ~ O HCI-C
~-OH
HO ~ 0~
~ S-CH3
~
A solution of 67 g (0,357 moles) of the amino acid.'HCl
(C.A. 51, 1643a, 1957) and 71.5 g (0.714 moies) of
triethylamine dissolved in 2.5 liters of acetonitrile is
cooled to 10 C and 47.6 g (0.354 moles) of isobutyl-
chloroformate added in one portion. This mixture
(Solution A) is stirred at 10 C for 1 ~our. Solution
B is made up by dissolving 97,7 9 (0.357 moles) of 7-Cl-
MTL (J. Med. Chem., 12-780, 1969, B. J. Magerlein and F.
Kagan) in a warm mixture of 1500 ml of acetone and 1500
ml of H20. Solution B is cooled to 30 C and added in one
~ortion to Solution A. The reaction is stirred at 25 C
for 18 hours and the acetone and acetnoitrile removed
under vacuum. The ~hite, mushy residue is filtered and
the crystalline material collected and dried to give 95 9
of pure product. Workup of the filtrate (chromatography)
`` 116486~
-20- 3809A
gave another 10 9 of product. The overall yield is 73~.
Anal. Calcd. for C17H2sClN205S: C, 50.42; H, 6.22; N,
6.g2; S, 7.92; Cl, 8.76.
. Found: C, 50.67; H, 6.40; N, 6.64; S, 7~90; Cl,
8.70.
~CHCl3 ~C, 1.0) ~ 293
PART II
~2Hs C2Hs CH3
~ /7-C1-MTL ~ C~H -CH H,O
HC1
~OH
\L__L/S-CH3
V ~ H
A mixture of 4.05 g (0.01 mole) of starting material,
40 ml of water, 60 ml of methanol, 1.0 ml of 37% HCl
and 8.0 g of PTO2 catalyst were reduced on a Parr
hydrogenator at 50 p.s.i. for 3 hours. Analysis of the
reaction mixture by TLC on silica gel plates in a system
composed of CHCl3:methanol (6:1) showed that, all of the
starting material was gone and that two more polar
materials were present in a ratio of about 1:1. The
reaction was filtered to remove the catalyst and the
filtrate concentrated under vacuum to give a,white
crystalline mush. This was filtered and the filtrate
saved. The white s~lid, which was the most polar of the
two products observed upon TEC of the reduction mixture,
was recrystallized from water to give the desired product,
U-57,930E, m.p. 222-224, in a yield of from 25 to 35~.
Anal. Calcd. for C17H32Cl2N205S: C, 45.63; H, 7.21; N,
1 16486'~
-21-` 3809A
6.26; S, 7.1~; Cl, 15.85.
Found: C, 45.77; H, 7.44; N, 6.39; S, 7.21; Cl,
16.17.
aH20 (C, 1,0) f 176
The absolute configuration and sterochemistry of V was
established by X-ray crystallography.
U-57,930E, tested in comparison with clindamycin,
has the following antimicrobial spectra:
Ta~le I
The Minimal Inhibitory Concentra~ion of U-57,930E
and Clindamycin Ys. Aerobic Bacteria.
MIC (~q/ml)
Organism UC Clindamycin U-57,930E
Staphylococcus 6685 >25 >2S
aureus 6686 . .S Z
6687 .025 .20
6688 >25 >25
6689 .OS .78
. 6690. .02S .20
6691 .10 .20
6692 >25 . . ~25
6693 .05 .78
6694 >25 >25
6695 .10 .39
66g6 .10 .39
6675 .05 .39
7~ .05 .10
746 <.05 .05
571 .20 .78
` 1~64864
-22- 3809A
570 .Z0 .39
Staphylococcus 719 .~0 .20epi dermi dis 3389 .10 .20
s
Streptococcus
faecalis 694 25 6.25
Streptococcus
pyogenes 152 <.012 <.012
Streptococcus
viridans 153 ~.012 .05
871 <.012 <.012
Diplococcus
pneumoniae I 41 <.012 ~.012
Diplococcus
pneumoniae II3213 <.012 <.012
Escherich;a c _ 45 50 >50
Proteus vul gari s 93 >50 >50
~leb-siella
pneumoniae 58 6.25 >50
Sa 1 morlel I a
schottmuelleri126 >50 >50
..... Ps~udomonas
aerugi nosa 95 ~50 >50
The procedure for the above test is as follows:
The Minimal Inhibitory Concentration (MIC's) o~ both
compounds Ys. aerobic bacteria is determined using a
` 116~86~
-23- 3809A
standard microplate broth-dilution method. 8rain Hedrt
Infusion (BHI - Di fco) broth medium is used, and the
plates are incubated at 37 C for 20 hrs.
S. aureus UC 6685-6696 are clinical isolates which
are resistant to one or more commercial antibiotics.
"UC" is a registered trademark of The Upjohn Company
Cùlture Collection. These cultures can be obtained from
The Upjohn Company in Kalamazoo, Michigan, upon request.
Table II
The Minimal Inhibitory Concentration of Olindamycin and
U-57930E vs. Gram-Positive and Gram-Negative Anaerobic
Bacteria
lS MICt~q/ml
Organism UC Clindamycin U-57930E
Bacteroides
fragilis 6513 0.06 0.12
,. ,~
cv 6428 0.06 0.25
6864 3.9 2.0
686Z 7.8 15.6
Bacteroides
thetaiotaomicron 6512 2.0 0.5
8acteroides
distasonis 6518 0.12 <0.03
8acteroides
melaninogenicus 6326 0.06 0.06
Cl os tridium
perfringens 247 0,06 0.l2
6509 0,06 0.12
C~ostridium
-
novyi B 6329 0.06 0.12
1 16486~
-2~- 3809A
Clostridium
tertium 6508 7.8 7.8
Clostridium
cadaveris 6510 <0.03 0.06
S
Clostridium
sordellii 6505 2.0 0.5
Clostr;dium
tentani 6521 <0.03 <0~03
Clostridium
botulinum A 6506 0.25 <0.03
Clostridium
bifermentans 6507 0.50 0.06
Clostridium~
difficile 6834 7.8 3.9
6857 250 125
6858 3-9 3 9
6860 500 500
6861 3.9 2.0
Propionibacterium
acnes 6564 0.06 0.12
6575 <0.03 0.06
Eubacterium
limosum 6515 2.0 2.0
.
Eubacterium
lentum 6522 0.50 1.0
Actinomyces
naeslundii 5920 0.25 0.25
-
8 6 4
-25- 3809A
Fusobacterium
nucleatum 6~16 0.12 0.12
6324 0.06 0.06
Fusobacterium
varium 6052 15.6 3.9
fusobacterium
necrophorum 6568 0.06 0.06
Peptococcus
asaccharolyticus 6214 0.50 0.25
Peptococcus
magnus 6258 0.06 0.06
Peptococcus
aerogenes 6319 <0.03 0.06
-
Peptostreptococcus
anaerobius 6321 0.12 0.12
The procedure for the above test is as follows:
Serial two-fold dilutions of drug are prepared in 1.0 ml
volumes of Schaedler 8roth, and 9.0 ml of molten (47 C)
Wilkens-Chalgren Agar Medium, infri, is added to the
antibiotic-supplemented broth. After mixing with the
antibiotic, the agar is poured into 100 mm x 20 mm petri
dishes. The dishes are allowed to stand on the bench
30 oYerni ght prior to inoculation.
Cultures are streaked on Wilkens-Chalgren Agar, and
grown for 48 hours at 37 C in a BBL Anaerobe Jar. Growth
~rom the plate is harvested, and a cell suspension is made
in Schaedler broth to e~ual the turbidity of a 0.5
~cFarland Standard ( 108 cellslml). fhe suspension is
pipetted into the ~ells of a Steers replicator, and
approx. 1-2 ~l is deliyered to the surface of the agar
1 164864
-26- 3809A
plates. After allowing a few minutes for the inoculum to
dry, the plates are placed in a BBL Anaerobe Jar (atmos-
phere o~ 85X N, 10% H, 5% C02) and incubated at 37 C for
72 hours.
The Minimal Inhibitory Concentration (MIC) is read
as the least amount of drug t~at inhibits growth. A very
faint film of growth, or ~3 colonies is considered
negative.
Wilkins-Chalgren Agar Medium
Dispense the following ingredients and dissolve in
1000 ml distilled water. The pH should be 7.0 - 7.2.
Trypticase 10 g
Gelysate 10 9
Yeast Extract 5 g
lS Glucose 1 Q
NaCl . 5 9
L-Arginine-Free Base 1 g
Py-ruvic Acid-Sodium Salt 1 g
Agar 15 g
Add Heme and ~Vitamin K1 solutions to yie7d
final concentrations of 5 llg/ml Hemin and 0.5
g/ml Ki
Autoclave at 121 C for 15 minutes aerobically.
Heme Sto~k - 0.5 g Hemin ~ 10 ml I N NaOH f 990
ml H2
Autoclave at 121 C for 12 minutes.
Add 10 ml stock per liter of medium.
Vitamin K Stock - .05 ml Vit. K1 solution t 20
ml 95% ethanol
Filter sterilize
Add 0.2 ml stock per liter of medium
The I.P. LD50 of U-57,930E in the mouse was found
to be 592 mg/kg. This value is the resulting mean of two
separate and identical LDso determinations. This value
is approximately 2 times the LD50 for clindamycin HCl.
The LD50 va1ue should be interpreted as indicating that
the acute I.P. toxicity of U-57,930E is approximately
one-half that of clindamycin HCl.
1 164864
-27- 3809A
In vivo : Mouse Protection Test
Or~anism & RouteClindamy~in U-57930A Ratio
S. aureus
Subcut. 5.7(4.2-7.8)* <<5
Oral 12.3(8.8-17.3) 1-5 ca.10
S. hemolyticus
Subcut. 2.3(1.6-3.3) .25 ca.10
Subcut. 3.3(2.~-4.2) .25(0.2-0.33) 13
Oral 12.3(10.2-14.8) 2.9 (2.0-4.1) 4.2
K. pneumoniae >320 >320
*CDso ' s as mg/kg
The procedure for the above test is as follows:
Mouse-protection Tests: Groups of 10 standard
laboratory mice (CF-l Mice) weighing 18-20 gm were
infected with approximately 100 LD50's of standardized
bacterial ce.ll suspensions which had been maintained
frozen at -170 C. Immediately before use, the suspen-
sions were thawed quickly and properly diluted. Infec-
tion was via the i,ntraperitoneal route.
Treatment of the infected groups was begun immediate-
ly and continued once per day for 4 days (first 24 hr
period = 1). Groups of untreated infected mice served as
virulence controls for the culture.
Seven days after the treatment regimen was begun the
surviving animals were sacrificed and the median protect-
ive dose of the antib;otic calculated on the basis of
mortality rates in the treatment groups. The median
protective dose and its 95% confidence interval were
calculated according to the method of Spearmen & Karber
as programmed on a 360 digital computer.
Also isolated from Example I, Part II, is compound
V A. This material is obtained as follows-
The filtrate which was saved from Part II was con-
centrated to dryness under vacuum, the residue converte^d
to its free base and chromatographed over silica gel using
1164864
-23- 3809A
CHCl3:methanol (6:1) as the eluting solYent. In this
manner the least polar material mentioned in Part II was
obtained. It was converted to its HCl salt and recrystal-
lized from acetone and water. This isomer is tentatively
being assigned structure V A.
C2H5
- CH~
~ HC-Cl
~ N J "C~ NH _ CH
HO
/ \
V A ~ -CH3
OH
Epimerization of the carbonyl function attached to
the piperidine ring of Y and V A may be accomplished by
methods wel~ known to those skilled in the art. The
trans isomers V B and V C produced by these epimeriza-
tions may be iso1ated by conventional procedures such ascrystallization or chromatography.
C2H5
.. ~
y ~ N J""~7 1 M Y B
C2H s
Y A eplmerize> ~ ~ VC
. H C~7-Cl-M~
Alternatively, V and V b may be hydrolized to give the amino
acids V D and V E which may then be epimerized by methods well
known to those skilled in the art to V F and V G, respectively.
The amino acids V E and Y G may be coupled with any of
~ 164864
-29- 3809A
the lineosaminides described earlier.
C2H5 C2H5
Hydrol;ze ~ epi.merize ~
> O O
~ N~ C-OH ~N ~ ~-OH
V D H V F H
C2Hs C2~5
V Hydrolize ~ epimerize ~
O > O
~ ~ ~-OH ~ N~ "'C-OH
lS H H
YE V G
The D-cis isomer (V A) of U-57,930E has an anti-
bacterial spectrum when tested on BHl broth as described
previously in Table 1.
Organism UC No.MIC (~g/ml)
_ aureus 76 250
570 1000
746 125
5. fecalis 694 >1000
S. pyoqenes 152 62.5
- _ pneumoniae 41 62.5
E. coli 45 ~1000
30 K. pneumoniae 58 >1000
_ schottmuelleri 126 >1000
Ps. aeruqinosa 95 >1000
. . .
Example 2 - Other Analogs of 7-Cl-MTL.
By following the procedures of Example 1, but
substituting the amino acid with the following amino
acids there are prepared the corresponding novel
1 ~64864
-30- 3809A
antibacter;ally-active analogs as their free bases or
acid addition salts. The latter can be prepared by
methods well-known to those skilled in the art.
Amino Acid Analo~
s
U-45,863
C-OH
o 1
_ C-OH
U-46,138
N
HO-C=O
U-46,137
O U-46,337
C-OH (Fast isomer on TLC-
N MeOH:CHCl 95:5. Run on
H silica gel plates~
O U-46,465
C-OH (Slow isomer on TLC)
N
l U-46,699
C-OH (Slow isomer on TLC,
I prepared from U-46,465)
CH3
- 1 16486~
-31- 3B09A
Hsca ~8 U-45,656
~N C-OH
C2H5
U-45,652
C-OH
C2Hs~L~ U-46,701
-OH
N
CH3
C2Hs~L_o U- 6 0, 481
C-OH
2 0 . H
~ U-44,469
. ~IN~-OH
CH2
CH2
OH
0
C - OH
~ ~L Cl U-45,657
1 164864
-32- 3809A
Example 3 - Analoqs of MTL
By following the procedures of Example 1, but
substituting the amino acid with the following amino
acids and substituting MTL (J. Am. Chem. Soc., 89-2448,
1967 W. Schroeder, B. 8annister and H. Hoeksema) for 7-
Cl-MTL, there are prepared the corresponding novel anti-
bacterially-active analogs:
Amino Acid Analogs
tO - U-46,136
l~ cR oH
C2H5
U-45,6~3
C-OH
U-60,493 (fast
~ i somer TLC)
2 5 ~H~-OH
,/~ U-60,492 ~`slow
¦ 1 8 i somer TLC)
N~C-OH
H
Example 4 r Analog~ of e~i`-MTL
3~ By following the procedures of Exampl e 1, but
substituting the amino acid with the following amino
acids, and substituting epi-MTL (J. Chem. Soc, Perkin I
,, .
, ~ t ~
` 116~864
33 3809A
1974, p. 3SO-B, Bannister) for 7-Cl-NTL there are prepared
the corresponding novel antibacterially-acti~.e analogs;
Amino d C i d hnaloq
.
~ o U-46, 135
~ ~C-O~
N
F2H5 . U-.45,65g
1~1 0
1 5
Cpd. A
., ''~1 .
N -OH
H
C2Hs
~ Cpd. 8.
- 25 ~ 1l
~ ~ C-OH
Ex~mple S ~ Analo~s o~ epii~7~Cl~tT~
3~ By following the procedures o~ Example 1, but
substituting the amino acid with the followi.ng amino
acids, and substituting epi-7-Cl-MTL for 7-Cl-MTL7 there
are prepared the correspond.ing novel antibacterially-
active analogs:
1 16486~
3~- 3809A
Amino Acid Analog
~ C-OH Cpd, C
C2Hs
~ 1I Cpd, D
~ C-OH
Cpd, E
N ~OH
H
C~2~s
Cpd~ F
-OH
H
Epi-7-Cl-MTL can be prepared b~ the procedure used
to prepare 7-Cl-MTL with the exception that the starting
material is epi-MTL instead of MTL,
Chemical and p~sical c~aracterization of most of
the compounds of Examples 2~5 are as follows.
c 35
1 164B64
- 3 5 - 38 O9A
C H N S "Cl ~D. Mp.
,
U-45,863 1 47.80 S.62 7.44 8.51 9.41 CHC13
2 47.76 5.54 7.35 8.76 9.34 +286 96-100
U-46,138 1 47.80 5.62 7.44 8.51 9.41 - -
2 47.30 5.74 6.91 8 ~ Sl 9.44
10 U-46,137 1 47.80 5.62 7.44 8.51 9.41 EtOH
2 47.58 5.74 7.56 8.53 9.49 t216 189-190C
U-46,136 1 50.26 6.19 7.82 8.95 MeOH
2 49.16 5.86 8.01 9.15 ~194 l9g-201
U-46,135 1 50.26 6.19 7.82 8.95 MeOH
2 50.50 6.19 7.98 9.27 ~269 97-100
U-46,337A 1 42.96 6.73 6.68 7.65 16.91 EtOH
20` 2 42.73 6.86 6.52 7.73 16.68 ~206 220-30
U-46,465E 1 47.05 7.11 7.32 8.38 9.26 EtOH
2 46.63 7.37 7.12 8.47 9.33 ~231 180-3
25U-46,699E 1 44.34 6.98 6.47 7.40 16.36 H20
2 44.79 7.24 6.25 7.36 16.42 ~172 229-234
U-45,656 1 50.42 6.22 6.92 7.92 8.76 CHC13
2 50.85 6.39 6.72 7.54 8.93 ~250
3~
U-45,652 1 50.42 6.22 6.92 7.92 8.76 CHC13
2 51.03 6.40 6.65 7.56 8.02 +273
U-45,653 1 52.83 6.78 7.25 8.30 - MeOH
2 53.83 7.08 7.39 8.14 - +203
U-45,659 1 52.83 6.78 7.25 8.30 - CHC13
2 52.77 6.70 7.34 8.55 - +295
1164864
' -3~- 3809A
U-46,701A 1 46.85 7.43 6.07 6.95 15.31
2 ~
U-44,469E 1 44.06 6.96 6.05 6.92 15.30
S 2 44.83 6.68 6.07 6.72 15.49 - - - -
U-45,657 1 43.80 4.90 6.81 7.80 17.24MeOH
2 43.52 4.93 6.82 7.82 17.41 +181 10S-130
1 - CALCD.
2 - FOUND'
Example 6'- Fusaric Ac~'d'Ami'de of'7-Chloro-MTL.
lS HgC ~
CH3
HgC4 ~ O HC-Cl
C - NH ~H
~ S-CH3 (U-55,581)
OH
By following the procedure of Example 1. ~ut substi-
tuting the amino acid with ~usaric acid, there is
obtained U-55,581,
Anal. Calcd. for C19H2~ClN205S: C, 52,7~; H, 6.75; N,
6.47; S, 7,41; Cl, 8.19.
Found: C, 52.15; H, 6.65; N, 6~36; S, 7,21; Cl,
7.94.
Fxample 7 - 4-Cis-n-Butyl-L-Pipecolic Acid Amide of
7-Cl-MTL or U-60,970E
116~864
37_ 3809A
C4HgC4Hg
C-7-Cl-MTL ~ C//O:HHCIHCl (X)HzO
H1 ~ ~
~H ~
~ S-CH3
OH
(C17H32Cl2N205S-(X)H20)
A mixture of 4.0 9 (0.0093 mole) of starting material,
40 ml of water, 40 ml of methanol, 2 ml of 37~ HCl, and
8.0 9. PtO2 catalyst were reduced on a Parr hydrogenator
at 50 psi for 18 hours. The reaction was fittered to
remove the catalyst and the filtrate concentrated under
vacuum to give an amber oil. The oil was dissolved in
20 ml. of a 2:1 solution of CHCl3 and methanol and
enough triethylamine added to neutralize the HCl pre-
sent. This solution was then chromatographed over silica
gel using a solvent system composed of CHCl3:methanol
(2:1). Two main product fractions are obtained. The
fractions containing the faster moving material were
pooled and evaporated under vacuum to give a white solid,
fraction A. The fractions containing the slower moving
material were pooled and evaporated under vacuum to give
a white solid, fraction B. Fraction B was dissolved in
a small amount of H20 and enough 37X HCl added to make
the pH 2. Crystallization occurred. The solid was
collected and recrystallized from H20 to give white
crystals of the desired product, U-60,970E, m.p. 224-
226 in a yield of 25-35%.
Anal. Calcd. for C17H32Cl2N205S: C, 47.99; H, 7.63; N~
5.89; S, 6.75; Cl, 14.92.
Found: C, 47.97; H, 7.42, N, 6.23; S, 6.90; Cl,
14.87
116~864
-38- 3809A
~leOH + 178 (C, 1.0)
CMR analysis supports the proposed structure.
The Minimal Inhibitory Concentration (MIC) in ~g/ml
of U-60,970E against various bacteria is as fo110ws:
Organism UC# MIC
_ aureus 76 0.125
570 0.25
746 0.062
_ faecalis 694 0.25
S. pyogenes 152 0.008
D. pneumoniae 41 0.016
_ coli 45 31.2
K. pneumoniae 58 1.8
S. schottmuelleri 126 31.2
Ps. aeru~inosa 95 >125
The test procedure is as disclosed in Example 1.
U-60,970E was also tested in vivo in standard lab-
oratory mice which were experimentally infected with
bacteria. The test was conducted in comparison with
U-57,930E. The following results show that U-60,970E
is significantly more active in viYo against D. pneumoniae
I and III than U-57,930E. Against S. aureus and S.
hemolyticus U-60,970E demonstrates essentially the same
activity as U-57,930E.
1 164864
-39- 3809A
_ _ .~ .
r~ r~
t un
I U~
r~
c
a~
~ C`J 0
O
a) ~ ~
_ O ~D ~.D d r~
1 0 ~ ~ r r~ O O j
E~ O 1~ c~ l ~ r~
c ~) O u~
c ~ O - O O O
O ~ _ _ _
Il~ ~ c~ l 1~ ~ c~
O ~ ~ u~ l
_ < 000
~0 _ _
o ~cô
a~ ~l ~ ~ co
., ~n ~ ~1 . u-~
~ C~
q~ 4~ . 1~ ~ , '
I I ~ 00 u~
c ~ ~
O . ~_ I~ I~ O O
~ ~ ~ ~ r~ a~ ~~
_ _ _
O _ _
Z O r~ a~ C~J ~) ~ ~
-a I 1~ ~
:E ~ co ., .
~0
a- ~ ~ ,_ ~
r~ r _~
O i
a~ a~
2 5 ~ C~
~___
o~ r~ ~ oo
O ~ <~ U~
~f) . . . . .
O ~_ r~ C~
_ _
_ I_
~ r~ c_ ~ I_
~ U ._ ~
c O r~ I_
3s ~ ~
1164864
.
-40- 3809A
Example 8 - 4-Cis-n-Butyl-D-Pipecolic Acid Amide of
7-Cl-MTL or U-61,734E
C4Hg C4Hg
C-7-Cl-MTL ~ ~ HC-Cl
N ` C-NH-CH
H
S-CH3
OH
(c17H32clN2oss- (X)H20)
Fraction A from the preceding experiment was converted to its HCl
salt in the same manner as described for fraction B. A
25-35% yield of product was obtained whose CMR spectrum
was essentially ident1cal to that obtained from fraction
B.
Example 9 -_Preparation of a Compound in Which the Amino-
acid Portion Contains a Heteroatom In a 5-
Membered Ring
I I Hydrolysis>
CH3-h ~ 1 C// CH~ OH
CH 3 CH3
+
7-Cl -MTL
CH3~ ~ ~ NH--CH
CH3 H ~ 0~
~CH3
OH
- ` 11648B4
-41- 3809A
The aminoacid ester (see C.A. 69 - 67282M) may be
hydroly~ed to the free acid by methods we11 known to
those skilled in the art ~acid or basic hydrolysis may
be used). It may be obtained in the form of the HCl
salt or the zwitterion. The coupling of the aminoacid
HCl with 7-Cl-MTL is accomplished in the same manner as
described in Example 1, except that 67.7 9. (0.357 moles)
of the aminoacid is used. After workup, as described in
Example 1, the crude product may be purified via chroma-
tography over silica gel and the product fractions com-
bined and converted to the HCl salt.
Example 10 - Preparation of a Compound in Which the Amino-
acid Portion Contains a Heteroatom in a 6-
Membered Ring
CH3 ~ Hydrolysis~ ~ /0
~N C-O-CH3 ~ C-OH
C~ Cl
CH3 +
CH3 ~ HC-Cl 7-Cl-MTL
N NH - CH
C1
The aminoacid ester (see C.A. 68 - 59465N) may be
hydrolyzed to the free acid by methods well known to
those skilled in the art (acid or basic hydrolysis may
be used). It may be obtained in the form of the HCl salt
~ 164~6~
-42- 3809A
or the zwitterion~ The coupling of the aminoacid-HCl
with 7-Cl-MTL is accomplished in the same manner as
described in Example 1 except that 103.6 9. (0.357 moles)
of the aminoacid is used. After workup, as described in
Example 1, the crude product may be purified via chroma-
tography over silica gel and the product fractions com-
bined and converted to the HCl salt.
Example 11 - 2-Phosphate Analo~s
The 2-phosphate analog of the compounds prepared in
Examples l-lOcan be prepared by procedures well-known to
those skilled in the art. By obvious appropriate modifi-
cation, the procedure disclosed in U.S. Patent 3,487,C68
may be used. Basically, any procedure would first
involve the protection of vulnerable groups by methods well-known
to those skilled in-the art-whieh would then be removed upon com-
pletion of the phosphorylation.
Example 12 - 2-Palmitate Analogs
The 2-palmitate analog of the compounds prepared in
Examples l-lOcan be prepared by procedures well-known to
those skilled in the art. By obvious appropriate modifi-
cation, the procedure disclosed in U.S. Patent 3,580,904
may be used. Basically, any procedure would first
involve the protection of vulnerable groups by methods well-known
to those skilled in the art which would then be removed upon com-
pletion of the acy.lation with palmitoyl chloride.
The Minimal Inhibitory Concentration (MIC's) of arepresentatiYe-num~er of the compounds prepared in
Examples 2-8 follows. The test procedure is as given
supra.
1 164864 3809A
~ ~ ~OOa~O 000~ ~ ~
1~ N ~) ~r, O O O O O O t~
~O ~/ ,_ C~.J t~J O O _ O O O l ~
.~ ~ ~ ~ . ~:
X- I _ _ _ ~ _ _ _ ._ _ _ _ ~
a~ o o o .--o o o o o o
~ ~ ~: X _ u~ In O c~ O O O O O O
~3_~_~ ~ C~J C~l A O O O O A A
CJ
_ _ _ _ _ _ _ _ _
. ~ . ._
O ~ C~J ~ ~ O O O O O O
I~ ~ ~ x ~ c~l c~l l o o o o o o t a~
~ ~ ~D tf~ O O O O O
~ , ~ ~ ~ ~ ~ '~ 0=~~ .
a~ _ _ _ _ _ _ _ _ __
1 1, 1 1
a I~J . I I I I I ~
I ~.) n ~ o o ol o o ol o ol 01 01 01 0
El ~ Df ~: o o l O 0 01 0 01 01 01 01 0
C ~ , O O 01 ~ O 01 0 01 01 01 01 U~
~ ~ U~ ~_ _ .-- ~ 1 _ ,--1 .
El ~ , ~ ~` l ~ I A ~ 1 1 A l l
.cl~c _ _t_ t I I I I
~I s l~ _~ ol ol ol o o I o ol ol ol ol o
q)l-o /~ ~ ol ol 01 0 o I O ol ol ol ol o o~
f ol ol01 0 O I O 01 01 01 01 o
E IL~7 . I ~ ~ I ~ --I ~ I --I ' - I --
~1~ ~I Al~I ~ ~ I ~` ~l Al Al Al A
Z~
ll l l l i ~
='lx I I I I I I I I I
1~ ~1~ ol ol ol o ol I o ol ol ol ol o
~ ~ ol ol ol o ol I o ol ol ol ol o
I ~ ~ ol ol ol ~ ol I o ol u~l ol ol ~ J
lU~'-1-1'-1 ~ I 1- 1-1
~ I I I I I I
I-D X ~ I I I I I I
'`'I'`JI'`'I~ ol 1o olololol' o=~
I'J I I l I I I l l I
l l l l l 1 1~
1 ~1 1 1 I 1 ~1 ~1
I 01 DI ~I I I
I ~1 ~1 ~rl I ~1 ol I ~1 ~1 1
I ~1 '~1 ~ Ul V~l ~ ~ ~ l o
U~ C J
I ~1 _1 ~I c~ ~ I Cl El ~I C
cn I <~ I I ~I E ~ E
1' 1 ~1 ~1 a)l E¦ ~ ¦ = ,~ a¦ l '--I
L , ~ ~ J o_Y l ~ I . c~ l
1 164864
44 3809A
a~ x o~ a:~--0 a ~ _ _ _ o s~ s_
^ C~ ~~ r~ I~ r~ ~ ~ o o o o O ~ E o o
~ ,- ,_ , ~ z o v~_ _ _ _ _ _ _ _
a~ ~ a~ o a~ o
U~ ~X ~ ~ ~ ~ f~ O O O O O o Q~
~o C3 ~ V . ,oo--~ o o o V 3 E ~
A ~ '.~~ O
r~ X o o o o o o o o o o c Q~
O o O u~ o o O O O O u~ ~ E ',--
~ z~ o o o c~ o o o o o o c~ "~ o
r~ J ~ f--r ~ r ~ r~ f-- ~
In ~4 _ _ _ _ A _ _ _ 1~ ~_ O
~r) _~ O O O O O O O O O O O
. _ O O O O O O O O O O O
~ ~ O O O O O O O O O O O
U`f_ ,_ r~ f--~ ~-- ~ f--f--,_ ,_ f--
a~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
S_ _ _ _ _ _
~) ~ ~ O O O O O O O O O O O
E ~ ,- ~ o o o o o o o o o o o
_ ~ r, ~ o fo _ o o o o o o o o
E ~ e5- ~, ~ ~ ~ ~ ~ A A ~ ~ ~:
~ ~ _ _ _ _ _ 0=~
: L r O g g O g g O o g 0 8
~ E ~D ~ O O O ~ O ~ O O O O U~ ~ ~
Zl ~ ~ ~ ~ ~ ~ _ ~ ___ _ I_
~ 0 x I I I .11."1 1~1 I
~ I I 1~-1 f~l fl ~1_'1 l
~ I I !q~ l I ~ ~o=~
~ % u~ ol I I I I I u~ ~_
ID ~. Nl I I Nl 01 1 01 01 0 01 0 N 0
Ct~ DI I I 1 01 1 01 01 0 01 0 ~O
~ ~ I I I I l I l l l ~
I I I 1^1 I^L^I^ ^I^--x
I I I I I I I I _I
l l l l l l l l N l
I I I '`'I I I I I ~
I I 1~1 o~l I I I c~ 'I ` ~1
ol ~ s
1 ~1 1 ~1 1 1 1 ~ a,i ~1
'-I ~1 '-I ,1 'I I ,...,1 ''I a~ , I ~ ~,1
JI v~l I crl I ~ Q~l O ,al
~1 ~1 ~1 ~ _ ~1 c ,
" 1 ,J,I ~ ~1 1 ,1 C I o El _ ol
~1 ~1 ~1 _1 ~)1 1 1 ~I c ~1 ~ El
a~ l a~l ol a~l I, I crl ~ ol s_ ~1
I I C I El ~1 1,--1 I a) --I u~ ~)1
a~l ~1 1 ol ~I c t.)l ~ cl
'I 'I ~ ~nl ~
.1 .1 .1 .1 ~1 1 .1 .1 . .1 V~.1
U~ I V I ~7 1 U~ I V~ I I LLJ I o_ ' ~ V~ o I
. . _
1 16~864
3809A
MIC in mcg~ml
U-Number and Structure
-
~ U-60,493 U-60~492U-60,481
S . ~L ~ R~X
_ .
S. aureus UC 76 1000 1000 2 0
. _ .
S. aureus UC 570 1000>1000 _ _ 3.9
1 S. aureus UC 7q6 250 250 2.0
O
S. hemolyticus UC 152 7.8 15.6 :~1.0 .
St. faecalis UC 694 tOOOjlOOO _31_.2 ,
. . __
S. lutea UC 130
E. coli UC 45 ,1000 >1000 500 -
.
P. vulgaris UC 93 >1000 >1000 1000
K. pneumoniae UC 58 >1000 >1000 250
. .
S Sc~ott~el ~ trf UC I ~ ; >1 000 >1 0001 000
Ps. aeruginosa UC ~5 >1000 >1000_ _>1000
O. pneumoniae UC 41 15.6 31.2
Fast Slow
isomer isomer
on T~C on TLC IR &
NMR OK
, O O
11 11
X=C-7-Cl-~TL' Y=C-MTL
R=C 2Hs
. . .
Since the compounds of the subject invention are
actiYe against various Gram-positive and ~ram-negative
microbes, they can be used in various environments to
inhibit such microbes. For example, they can be used as
disin~ectants to inhibit S. aureus on washed and stacked
food utensils contaminated with this bacterium. They al50
can be used as disinfec,tants on various dental and medical
- equipment contaminated with S. aureus. Further, the
compounds of the inYention can be used as bacteriostatic
rinses for laundered clothes, and for impregnating papers
1 16~86~
-~t6 3~09A
and fabric~; and, they are also useful for suppressing the
growth of sensitive organisms in plate assays and other
microbiological media.
The compounds of the subJect invention exist in the
protonated or non-protonated forms according to the pH of
the environment. When the protonated form is intended,
the compounds exist as pharmaceutically-acceptable acid-
addition salts, and when the non-protonated form is
intended, the compounds exist as the free base, The free
bases can be converted to stable acid~addition salts by
neutralizing the free ~ase with the appropriate acid,
about pH 7.0, and advantaQeously to about pH 2 to pH 6.
Suitable acids for this purpose include hydrochloric,
sulfuric, phosphoric, thiocyanic, fluosilicic, hexafluoro-
arsenic, hexafluorophosphoric, acetic, succinic, citric,lactic, maleic, fumaric, pamoic, cholic, palmitic, ~ucic,
camphoric, glutaric, glycolic, phthalic, tartaric, lauric,
stearic, salicylic, 3-phenylsalicylic, 5-phenylsalicylic,
3-methylglutaric, o.rthosulfobenzoic, cyclohexanesulfamic,
cyclopentanepropionic, 1,2-cyclohexanedicarboxylic, 4-
cyclohexanecarboxylic, octadecenylsuccinic, octenyl- -
succinic, methanesulfonic, helianthic, Reinecke's,
dimethyldithiocarbamic, hexadecylsulfamic, octadecyl-
sulfamlc, sor~ic, monochloroacetic, undecylenic, 4'-
hydroxyazobenzene-4-sulfonic, octadecylsulfuric, picric,
benzoic, cinnamic, and like acids.
The acid-addition salts can be used for the same
purposes as the free base or they can be employed to up-
grade the same. For example, the free base can be con-
verted to a water-insoluble salt, such as the picrate,
which can be subjected to purification procedures, for
example, solvent extractions and washings,.chromatography,
fractional liquid-liquid extractions, and crystallization,
and then used to regenerate the free base form by treat-
ment with alkali or to make a different salt by meta-
thesisO Or the free base can be converted to a water-
solub1e salt, such as the hydrochloride or sulfate, and
116~86~
` -47- 3809A
the aqueous solution of the salt extracted with various
water-imm;scibte solvents before regenerating the free
base form by treatment of the thus-extracted acid solu-
t:ion, or converted to another salt by metathesis.
In addition to the antibacterial uses, disclosed
above, the free bases can be used as buffers or as ant-
acids, The thiocyanic acid addition salt when condensed
wit~ formaldehyde forms resinous materials useful as
pickling inhibitors according to U.S, patent Nos.
2,425,320 and 2,606,155. T~e free bases also ma~e good
vehicles for toxic acids. For example, the fluosilicic
acid addition salts are useful as mot~proofing agents
according to U.S, patent Nos. 1,915,334 and 2,075,359 and
the hexafluoroarsenic acid and hexafluorop~osphoric acid
addition salts are useful as parasiticides according to
U.S. Patent Nos. 3,122,536 and 3,122,552,
The compounds of the su~ject invention are useful as
antibacteria~ agents in sui:ta~le compositions, These
compositi;ons are prefera~l~ presented for administration
to humans and animals in unit dosage forms, such as
tablets, capsules, pills, powders, granules, sterile
parenteral solutions or suspensi:ons, and oral solutions
or suspensions, aad oil-water emulsions containing suit-
able quantities of the active compound in t~e form of the
free base, or its pharmacologicall~ accepta61e salts,
For oral administration, either solid or flu;d unit
dosa~e ~orms can be prepared. For preparing soljd compo-
sitions such as ta61ets, t~e principal active ingredient
is mixed with conventional ingredients such as talc,
magnesium stearate, dicalcium phosphate, magnesium
aluminum silicate, calcium sulfate, starch, lactose,
acacia, methylcellulose, and functionally similar mater-
ials as pharmaceutical diluents or carriers The tablets
can be laminated or otherwise compounded to provide a
dosage form affording the advantage of prolonged or
delayed action or predeterm~ned successiYe action of
the enclosed medication. For example, the tablet can
1 16486~
4~ 3809A
comprise an inner dosage and an outer dosaye component,
the latter being in the form of an envelope over the
former. The two components can be separated by an
enteric layer which serves to resist disintegration in
the stomach and permits the inner component to pass intact
into the duodenum or to be delayed in re1ease. A variety
of materials can be used for such enteric layers or coat-
ings, such materials including a number of polymeric acids
or mixture of polymeric acids with such materials as
shellac, cetyl alcohol, cellulose acetate phthalate,
styrene maleic acid copol~mer and the like. Alternatively,
the two component system can be utilized for preparing
tablets containing two or more incompatible active
ingredients. Wafers are prepared in the same manner as
tablets, di~fering only in shape and the inclusion of
sucrose or other sweetener and flavor. In their simplest
embodiment, capsules, like tablets, are prepared by mix-
ing the compound of the formulation with an inert pharma-
ceutical diluent and filling and mixture into a hard
gelatin capsule of appropriate size. rn another embodi-
ment, capsules are prepared by filling hard gelatin cap-
sules with polymeric acid coated ~eads containing the
active compound. Soft gelatin capsules are prepared by
machine encapsulation of a slurr~ of the active compound
with an acceptable vegetable oil, light liquid petrolatum
or other inert oil.
Fluid unit dosage forms for oral administration such
as syrups, elixirs, and suspensions can be prepared, T~e
water-soluble forms of the active compound can be dis-
solved in an aqueous vehicle together with sugar,aromatic flavoring agents and preservatives to form a
syrup. An elixir is prepared by using a h~dro-alcoholic
(ethanol) vehicle with suitable sweeteners such as sucrose
together with an aromatic flavoring agent. Suspensions
can be prepared of the insoluble forms with a syrup
vehicle with the aid of a suspending agent such as acacia,
tragacanth, methylcellulose and the li~e.
1 16~864
` -49- 38~9A
Topical ointments can be prepared by dispersing the
active compound in a suitable ointment base such as
petrolatum, lanolin, polyethylene glycols, mixtures there-
of, and the like Advantageously, the compound is finely
divided by means of a colloid mill utilizing light liquid
petrolatum as a levigating agent prior to dispersing in
the ointment base. Topical creams and lotions are pre-
pared by dispers;ng the compound in the oil phase prior
to the emulsification of the oil phase in water.
For parenteral administration, fluid unit dosage
- forms are prepared utilizing the active compound and a
sterile vehicle, water being preferred. The active com-
pound, depending on the form and concentration used, can
be either suspended or dissolved in the vehicle. In pre-
lS paring solutions, a water-soluble ~orm of the active
compound can be dissolved in water for injection and
filter steril.ized before filling into a suitable vial or
ampul and sealing. Advantageousl~ adjuvants such as a
local anesthetic, preservative and buffering agents can
be disso1ved in the vehicle. To enhance the stability,
the composition can be fro7en after filling into the vial
and the water removed under vacuum. T~e dry lyophilized
powder is then sealed in the vial and an accompanying vial
of water for injection is supplied to reconstitute the
powder prior to use. Parenteral suspensions are prepared
in substantially the same manner except that the active
compound is suspended in the vehicle instead of being
dissolYed and sterilization cannot ~e accomplished by
filtration. The active compound can be sterilized by
3~ exposure to ethylene oxide before suspending the sterile
vehicle. Advantageously, a surfactant or wetting agent
is included in the composition to facilitate uniform
distribution of the active compound.
The term unit dosage form as used in t~e specifica-
tion and claims refers to physicall~ discrete units su;t-
able as unitary dosages for human subjects and animals,
each unit containing a predetermined quantity of active
1 164864
-50- 3809A
material calculated to produce the desired therapeutic
e~fect in association with t~e required pharmaceutical
diluent, carrier or vehicle. The specifications for the
novel unit dosage forms of t~is invention are dictated
S by and directly dependent on (a) the unigue characteris-
tics of the active material and the particular thera-
peutic effect to be achieved, and (b) the limitations
inherent in the art of compounding such an active material
for therapeutic use i.n humans and animals, as disclosed in
detail in th;s specification, these ~eing features of the
present invention. Examples of suita~le unit dosage forms
in accord with thi~ invention are ta~lets, capsules, pills,
troches, supposito.ries, powder packets~ granules, wafers,
cachets, teaspoonfuls, ta~lespoonfuls, dropperfuls, ampuls,
vials, segregated.multiples of any of the foregoing, and
other forms as herein descri~ed,
In addition to the administration of the active com-
pound as the princ.ipal active ingredient of compositions
for the treatment of the conditions descri~ed herein, the
said compound can ~e included with other types of com-
pounds to o6tain advantageous com6inations of properties,
Such combina.tions i.nclude the active compound with anti-
~iotics suc~ as spectinomycin, chloramphenicol, novo-
~iocin, dih~dranovo~iocin, tetracy~clines (e,g., tetra-
cycline, oxytetracycline and c~lortetracycline) ? peni-
ci11ins, erythromycin ? kanam~cin, streptom~cin, neomycin,
polymyxin, bacitracin, nystatin, filipin, fuma~illin and
endom~cin to hroaden the ~acterial spectrum of the compo-
sition and for synergistic action against particular
bacteria.;. steroids having anti-inflammatory actiYity such
as h~drocortisone, prednisolone, 6-meth~lprednisolone,
6a-fluoroprednisolone and the like; analgesics such as
aspirin, sodium salicylate (acetylsalicylic acid)-
anhydride, N-acetyl-p-aminophenyl and salicylamide;
antihistamines, such as chlorpheniramine maieate,
diphenylhydramine, promethazine, pyrathiazine, and the
like; sulfas, such as sulfadiazine, sulfamethazine,
1 16'1~
-51- 3~09A
sulfamerazine sulfacetamide, sulfadimethyloxazole,
sulfamethizole, and the like; antifungals, such as
undecylenic acid, sodium propionate, salicylanilide,
sodium caprylate, and hexetidine; and the vitamins.
The dosage of the active compound for treatment
depends on route of administration; the age, weight, and
condition of the patient; and the particular disease to
be treated. A dosage schedule of from about 15 to 500
mg., 1 to 4 times daily (every six hours), embraces the
effective range for the treatment of most conditions for
which the compositions are effective. For children, the
dosage is calculated on the basis of 15 to 30 mg,/kg./day
to be administered every six hours.
The act;ve compound is compounded with a suita~le
pharmaceutical carrier in unit dosage form for convenient
and effective administration. In the preferred embodi-
ments of this invention, the dosage units contain the
compound in. 15, 30, 50, 125, 250 and 500 mg. amounts for
systemic treatment; in 0.25, 0.5, 1, 2 and 5% amounts for
topical or localized treatment; and 5 to 65X w/v for
parenteral treatment. The dosage of compositions contain-
ing the active compound and one or more other active
ingredients is to ~e determined with reference to the
usual dosage of each such ingredient.
The following examples are illustrative of the best
mode contemplated by the inventor for carrying out his
invention and are not to be construed as limiting.
The examples use U-57,930E or U-60,970E as the active compound,
but it should be understood that this is only exemplary -
of the other active compounds of the subject invention.
As distinguished from the previous examples, the follow-
ing are Composition ExamplesO
Composition Example 1
Capsules
One thousand two-piece hard gelatin capsules for ora
use, each containing 250 mg. of U-57,930E or U-60,970E are prepared
from the following types and amounts of materials:
~ 16486'1
-52- 3809A
Gm.
U-57,930E or U-60,970E 250
Corn starch lOO
Talc 75
Magnesium stearate 25
The materials are thoroughly mixed and then encap-
sulated in the usual manner.
The foregoi ng capsules are useful for the systemic
treatment of infection in adult humans by the oral
lO administrat;on of l capsule every 4 hours.
Using the procedure above, capsules are similarly
prepared containing U-57,930E or U-60,970E in 15, 30, 50, 125 and 500
mg. amounts by substituting lS, 30, 5û, 125, and 500 gm
of U-57,930E or U-60,970E for the 250 gm used above.
.
15 Composition Example_2
Capsules
One thousand two-piece ~ard gelatin capsules for oral
use, each containing 200 mg. of U-57,930E or U-60,970E and 250 mg. of
tetracycline hydrochloride, are prepared from the follow-
ing types and amounts of ingredients:
Gm,
U-57,930E or U-60,970E 200
Tetracycline hydrochloride25
Talc 75
Magnesium stearate 25
The ingredients are thoroughl~ mixed and then encap-
sulated in the usùal manner,
The foregoing capsules are use~ul for the systemic
treatment of infection in adu1 t humans ~y the oral
3Q administration of 1 capsule every~ 6 hours,
Using the procedure above, capsules are similarly
prepared containing U-57,930E or U-60,970E and each of the following
antibiotics ~n -p~ace- of- i;etracycline by substituting 250
gm. of such other antibiotic for tetracycline: chlor-
35 amphenicol, oxytetracycline, chlortetracycl;ne, fumagillin,erythromycin, streptomycin, dihydronovobiocin and novo-
biocin. When a penicillin, such as potassium penicillin
li6~4864
-53-
3809A
G, is to be used in place of tetracycline, 250,000 units
per capsule is employed.
Such combination products are useful for the system-
ic treatment of mixed infections in adult humans by the
5 oral administration of 1 capsule every 6 hours.
Composition Example 3
Tablets
One thousand tablets for oral use, each containing
500 mg. ~f U-57,930E or U-60,970E are prePared from the following
types and amounts of materi`als:
Gm.
U-57,930E or U-60,970E 500
Lactose 125
Corn starch 65
Magnesium stearate 25
Light liquid petrolatum 3
The ingredients are thoroughly mixed and slugged.
The slugs ar'e broken down by forcing through a number
sixteen screen, The resulting granules are then com-
pressed into tablets, each tablet containing 500 mg. of
U-57,930E or U-60,970E
The foregoing tablets are useful for systemic treat
ment of infections in adult humans by oral administration
of 1 tablet every 4 hours.
Using the above procedure, except for reducing the
amount of U-57,930E or U-60,970E to 250 gm., tablets containing 250
mg. of U-57,930E or U-60,970E are prepared.
Composition Example 4
- Tablets
One thousand oral tablets, each containing 250 mg.
of U-57,930E or U-60,970E, and a total of 250 mg. (83.3 mg. each) of
sulfadiaz;ne, sulfamerazine, and sulfamethazine, are pre-
pared from the fol'lowi'ng types'~a~nd'''amounts of materials:
~m.
U-57,930E or U-60,970E 250
Sulfadiazine 83.3
Sul~amerazine 83.3
1 164864
.
-5~- 3809A
Sulfamethazine 83.3
Lactose 50
Corn starch 50
Calcium stearate 25
Light liquid petrolatum 5
The intredients are thoroughly mixed and slugged.
The slubs are broken down by forcing through a number
sixteen screen. The resulting granu1es are then com-
pressed into tablets, each containing 250 mg. of U-57,930E
or U-60,970E and a total of 250 mg. (83.3 mg. each) of sulfadiazine,
sulfamerazine, and sulfamethazine.
The foregoing tablets are useful for systemic
treatment of infections by the oral administration of 4
tablets first and then 1 every six hours.
For the treatment of urinary infections, the triple
sulfas in the above formulation is advantageously
replaced by 250 gm. of sulfamethylthiadiazole or 250 gm.
of sulfaceta~ide.
Composition Example 5
Oral Syrup
One thousand cc. of an aqueous suspension for oral
use, containing in each 5 cc. dose, one-half gram of total
sulfas and 250 mg. of U-57,930E or U-60,970E is prepared from the
following types and amounts of ingredients:
Gm.
U-57,930Eor U-6Q,970E 50
Sulfadiazine 33.3
Sulfamerazine 33.3
Sulfamethazine 33.3
Citric acid 2
Benzoic acid
Sucrose 700
Tragacanth 5
Lemon oll 2 cc.
Deionized water, q.s. 1,000 cc.
The citric acid, benzoic acid, sucrose, tragacanth,
and lemon oil are dispersed in sufficient water to make
1 16~8~4
-55- 3809A
850 cc. of solution. The U-5J,930E or U-60,970E and finely powdered
sulfas are stirred into the syrùp until uniformly dis-
tributed. Sufficient water is added to make 1 ,000 cc.
The composition so prepared is useful in the systemic
5 treatment of pneumonia in adult humans at a dose of 1
teaspoonful 4 times a day.
Composition ExamPle 6
Parenteral Solution
A sterile aqueous solution for intramuscular use,
containing in 1 cc. 200 mg. of U-57,930E or U-60,970E is prepared fro~ l
the following types and amounts of materials:
Gm.
U-57,930E or U-60,970E 200
Lidocaine hydrochloride 4
Methyl paraben 2.5
Water for injection, q.s. 1,000 ccO
The ingredients are dissolved in water and the solu-
tion sterilized ~y filtration. The sterile solution is
filled into vials and the vials sealed.
20 ComPosition Example 7
Parenteral Preparation
A sterile aqueous solution for intramuscular use,
containing in 1 cc. 200 mg. of U-57,930E and U-60,970E and 400 mg. of
spectinomycin sulfate, is prepared from the following
types and amounts of ingredients:
Gm.
U-57,930E or U-60?970E 200
Spectinomycin sulfate 400
Lactose 50
Water for injection, q.s. 1,000 cc.
The U-57,930E or U-60,970E, spectinomycin sulfate and lactose
are dissolved in the water and the solution sterilized by
filtration. The sterile solution, in the amount of 2 cc.,
is aseptically filled into sterile vials and frozen. The
35 water is removed under high vacuum and the vials contain-
ing the lyophilized powder are sealed. Just prior to use,
sufficient sterile water for injection to make 2 cc. of
- 1 L64864
-~6- 3809A
solution is added to the vial.
Composition Example 8
Topical Ointment
One thousand gm. of 0.25% ointment is prepared from
5 the following types and amounts of ingredients:
Gm.
U-57,930E or U-60,970E 2.5
Zinc oxide 50
Calamine 50
Liquid petrolatum (heavy) 250
Wool fat 200
White petrolatum, q.s. 1,000 gm.
The white petrolatum and wool fat are melted and 100
gm. of liquid petrolatum added thereto. The U-57,930E or U-60,970E,
zinc oxide and calamine are added to the remaining liquid
petrolatum and the mixture milled until the powders are
finely divided and uniformly dispersed. The powder mix-
ture is stirred into the white petrolatum mixture and
stirring continued until the ointment congeals.
The foregoing ointment is usefully applied topically
to the skin of mammals for the treatment of infection.
The foregoing composition can be prepared by omitting
the zinc oxide and calamine.
following the procedure above, ointments are similarly
prepared cont~ining U-57,930E or U-60,970E in 0.5, 1, 2, and 5%
amounts by substituting S, 10, 20 and 50 gm. of U-57,930E or
U-60,970E for the 2.5 gm. used above.
Composi tion Example 9
Cream
One thousand gm. of a vaginal cream are prepared from
the following types and amounts of ingredients:
Gm.
U-57 ,930E or U-60,970E 50
Tegacid Regular1 150
Spermaceti 100
Propylene glycol 50
Polysorbate 80 5
1164864
-57- 3~09A
,~lethylparaben
Deionized water, q.s. 1,000 grn.
' Sel f-emulsifying glyceryt monostearate from
Goldschmidt Chemical Corporation, New York, N.Y.
The Tegacid and spermaceti are melted together at a
temperature of 70-80 C. The methylparaben is dissolved
in about 500 gm. of water and the propylene glycol,
Polysorbate 80, and U-57,930E or U-60,970E are added in turn, main-
taining a temperature of 75-80 C. The methylparaben mixture
is added slowly to the Tegacid and spermaceti melt, with
constant stirring. The addition is continued for at
least 30 m;nutes with continued stirring until the
temperature has dropped to 40-4S C. The p~ of the final
cream is adjusted to 3.5 by incorporating 2.5 gm. of
citric acid and O.Z g. of dibasic sodium phosphate dis-
solved in about 50 gm. of water. Finally, sufficient
water is added to bring the final weight to 1 ,000 gm. and
the preparation stirred to maintain homogeneity until
cooled and congealed.
The forégoing composition is useful for the treat-
ment of vaginal infections in humans.
Composition Example 10
Ointrnent, Ophthalmic
One thousand gm. of an ophthalmic ointment containing
0.5% U-57,930É or U-60,970E are prepared from the following types and
amounts of ingredients:
Gm.
U-57,930E or U-60,970E 5
Bacitracin 12.2
Polymyxin B sulfate (10,000
units/mg.)
Light liquid petrolatum 250
Wool fat 200
White petrolatum, q.sO 1~000 gm.
The solid ingredients are finely divided by means of
an air micronizer and added to the light liquid petrolatum.
The m;xture is passed through a colloid mill to uniformly
- ` 1 16~864
-5~~ 3~09A
distribute the micronized par~icles. The wool fat and
white petrolatum are melted together, strained, and the
temperature adjusted to 45-50 C. The liquid petrolatum
slurry is added and the ointment stirred until congealed.
Suitably the ointment is packaged in one dram ophthalmic
tubes.
The foregoing ointment is usefully applied to the
eye for treatment of localized infection in humans and
other animals
Advantageously the foregoing composition can contain
S gm. (0.5~) of methylprednisolone for the treatment of
inflammation, and, alternatively, the bacitracin and
polymyxin 8 sul~ate can be omitted.
Composition Example 11
Eye-Ear Drops
One thousand cc. of a sterile aqueous solution for
eye or ear use containing ~O mg. ~f U-57,930E or U-6Q,970E and S mg.
of methylprednisolone in each cc. is prepared from the
following types and amounts of ingredients:
Gm.
U-57,930E or U-60,970E 10
Methylprednisolone phosphate sodium 5
Sodium citrate 4.5
Sodium bisulfite
Polyethytene glycol 4000 120
Myristyl-y-picolinium chloride 0.2
Polyvinylpyrrolidone
Deionized water, q.s. ad 1000 cc.
The ingredients are dissolved in the water and the
resulting solution is sterilized by filtration. The
solution is aseptically filled into sterile dropper
containers~
The composition so prepared is useful in the topical
treatment of inflammation and infection of the eye and ear
as well as other sensitive tissues of the animal body.
Composition Example 12
Troches
1 164864
-59- 3809A
Ten thousand troches are prepared from the following
types and amounts of ingredients:
Gm.
U-57,930E or U-60,970E 100
Neomycin sul fate 50
Polymyxin B sulfate (10,000
units/mg.)
Ethyl aminobenzoate 50
Calcium stearate 150
Powdered sucrose, q.s. 5,000 gm.
The powdered materials are mixed thoroughly and then
compressed into half gram troches following the usual
techniques for the preparation of compressed tablets.
The troches are held in the mouth and allowed to
dissolve slowly to provide treatment for the mouth and
throat o~ humans.
Composition Example 13
~ Suppository, Rectal
One thousand suppositories, each weighing 2.5 gm. and
containing 100 mg. of U-57,930E or U-60,970E are prepared from the
following types and amounts of ingredients:
Gm.
U-57,930E or U-60,970E 100
Polymyxin B sulfate (10,000
units/mg.) 1.25
Methylprednisolone
Ethyl aminobenzoate 75
Zinc oxide 62.5
Propylene glycol 162.5
Polyethylene glycol 4,00û q.s.
2,500 gm.
The U-57,930E or U-60,9ioE, polymyxin B sulfate, ~nethylpredniso-
lone, ethyl aminobenzoate, and zinc oxide are added to
the propylene glycol and the mixture milled until the
35 powders are finely divided and uniformly dispersed. The
polyethylene glycol 4000 is melted and the propylene
glycol dispersion added slowly with st;rring. The
1 164~64
- -60- 3809A
suspension is poured into unchllled molds at 40 C.
The composition ;s allowed to cool and solidify and
then removed from the mold and each suppository foil
wrapped.
The foregoing suppositories are inserted rectally for
local treatment of inflammation and infection.
Alternatively, the foregoing composition can be pre-
pared omitting the steroid.
Composition Example 14
Mast;tis Ointment
One thousand gm. of an ointment for the treatment of
mastitis in dairy cattle is prepared from the following
types and amounts of ingredients:
Gm.
lS U-57,930E or U-60,970E 25
Methylprednisolone acetate O.S
Light liquid petrolatum 300
Chlo-robutanol, anhydrous 5
Polysorbate 80 5
2% Aluminum monostearate-
peanut oil gel 400
White petrolatum, q.s. 1000 gm.
The U-57,930E or U-60,970E and methylprednisolone acetate are
milled with the lig~lt liquid petrolatum until finely
divided and uniformly dispersed. The chlorobutanol.
polysorbate 80, peanut oil gel and white petrolatum are
heated to 120 F. to form a melt and the liquid petrolatum
dispersion stirred in. With continued stirring, the dis-
persion is allowed to cool (and congeal) to room tempera-
ture and is filled into disposable mastitis syringes in10 gm. doses.
Composition Example 15
Animal Feed
One thousand gm. of a feed mix is prepared from the
following types and amounts of ingredients:
Gm.
U-57,930E or U-60,970E 10
1 16~864
-61- 3~09A
Soybean meal 400
Fish meal 400
Wheat germ oil S0
Sorghum molasses 140
The ingredients are mixed together and pressed into
pellets. The composition can be fed to laboratory
animals, i.e., rats, mice, guinea p;gs, and hamsters for
prophylaxis during shipping.
For other animals such as poultry, e.g., chickens,
ducks, turkeys, and geese, the composition can be added
to the animal's regular feed in an amount calculated to
give the desired dose of U-57,930Eor U-60,970E.
Composition Example 16
Following the procedure of each of the preceding
Composition Examples 1-15, inclusive, each antibacterially-
active compound of the subject invention is substituted in
an equivalent amount for the U-57,930E or U-60,970E shown in the
example to pPovide therapeutic properties.
Similarly, each of the above free base compounds can
be used in the form of a pharmaceutically (or pharmaco-
logically) acceptable acid addition salt, e.g., hydro-
chloride, sulfate, nitrate, phosphate, citrate, lactate,
acetate, tartrate and succinate.
Further, the 2-phosphate and/or 2-palmitate of each
of the above antibacterially-active invention compounds
can be substituted as the active ingredient to provide
compositions having therapeutic properties.
