Note: Descriptions are shown in the official language in which they were submitted.
~ ~ 6~3~
This application is the third of three divisional
ap~lications of copending application serial no. 362,485,
filed October 16, 1980.
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 3,496,1630 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 thereofD
Lincomycin has the following structural formula
CH3
i ÇH3
HO-~-H
C -N ~
~0~ H ~H
H ~ S~H3
~ OH
Clindamycin has the following structural formula
cb/
~.~65~
CH3
CH3
N \ IH
- N - ~
H H ~ H0~ ~H
SCH3
H QH
1 0 ~
BRIEF SUMMARY OF TH_NVE~NTION
This application relates to novel
and useful compounds of the general formula-
: ~ D ~ 1
. /
- R2 - ~ I
~: : I VII
E A
B ~
~: 2~0 :~ wherein A, B, D and E are nitrogen/ Cl ~ N-, oxy~en, 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
.
al~yl portion is from 1 to 8 carbon atoms, and isomeric
forms thereof, cycloalkyl~and substituted cycloalkyl,
: substitu~ed oxygen, substituted nitrogenl halogen, phenyl
~ and substituted phenyl or -(CH2)m-OH, -(CH2)m-NR4~51 and
: ~: ::
; ~:
a~
':
:~ ' , ~ ' ' ,
,
~ 6~
isomeric forms thereof, wherein-m is an integer of from
1 to 8, and R4 and R5 are hydrogen or al~yl of from 1
to 8 car~on 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:
--C--X
and X is the amino function of 7(R)-hydroxy-methyl l-thio-
~-lincosamlnide, 7(S)-hydroxy-methyl l-thio-~-lincosaminide,
7(S)-halo-methyl l-thio-~-lincosaminide, 7(R)-halo-methyl
l-thio-a-lincosaminide, 7(S)-methoxy-methyl l-thio-~-
lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl l-thio-
~-lincosaminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl
l-thio-~-lincosaminide or 7-deoxy-7(S)-(3-hydroxypropylthio)-
methyl l-thio-~-lincosaminide.
This application also relates to novel and useful
compounds, which are claimed in the above noted parent
application, of the general formula:
R2
N
: R3 . -
:
wherein Rl, which can be singly or multiply substituted
in the 2, 3, 4, 5, 6, 7, 8 or 9 position of the ring not
already substituted by R2, is as defined above; wherein R2,
mab/~ b
.
~ ~6 ~3~ ~
which can be singly substituted in an~ position of the ring
not already substituted by Rl, is as defined above; wherein
R3 is hydrogen, methyl, ethyl or C2H50H, and wherein n
is an i.nteger oE from 1 to 40
This application Eurther relates to novel and use-
ful compounds, which are cla~ .in a first diyisional application,
serial no. 414y643, filed Nove~ber 1, 1982, of the general formula;
Rl
1 ~ ~2
~ N /
wherein Rl, which can be s.ingly 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 ~1~ is as defined aboveO
This application still further relates to novel
and useful compounds/ which are claiméd in:a second diYisional applica
~ tion~ serial noO 414,644? filed Nove~ber 1, 1382~ of~the general formula:
: R~ ~ R
\ B ~
wherein A, B 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
,
. - 4 -
mabj
5~
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:
~H2)r$R
III
R~
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 formula:
~ R
~: : ~R2
~ ~ N Iy
:~ .
wherein Rl and R2 are as defined immediately above.
The pharmaceuticall~ acceptable acid addition salts of the
;~ above noted compounds including the 2-phosphates and 2-
palmitates, wherein the substituent is attached to the
oxygen atom at the 2 position of the sugar ring of the
above compounds, are also described and claimed in this
~ 5 --
~ ., ~ .,l )
and the above noted related applications.
The synthesis of the novel analogs described
herein can be shown in exemplary form as follows:
C Hs ~ H ~j~ O HF
- G-OH* ~H~C~
HO~O H~O,~O,~
~5-C ~-cH3
C2~s
.. C~3
Hl~-Cl
~N C-N~ÇH
H HO~O
I~ OH.
S-~H3
n~
The wavy lines denote either the D-cis or L-cis isomer.
An alternate procedure which may be used to
synthesize the novel analogs described herein can be shown
in exemplary form as follows:
~2H5 . C2H5
/ ~ 7 Cl-MTL ~ ~C-7-Cl-MTL
~:~ H H
'
The wavy line denotes either the D-cis, L-cis, D-trans
or L-trans structures.
- - 6 -
mab/~ >
. .
i53~j
The L-cis structure
C2H5
,1~ f H3
~HCl
N C--NH IH ~ XH 2(~
HO~ ~
~S.CH3 y
OH
has been shown to be 5 to 10 times more active than elinda-
mycin against S. aure-us and-S. hemolyticus in laboratory
mice.
An isomer of V may be isolated from the above
reaction and is presumed to be the D-eis eompound, VA.
The D-cis structure is not as potent an antibacterial
a~ent as the L-cis compound.
~;20~
C2H5
C H 3
N J~c - N H--C H
H
H~
0~
DETAILED DESCRIPTION OF THE INVENTION
Upon reacting an amino aeid of the formula
` - 6a -
-7- 3809A
~pol~
N
wherein R1, which can be singly or multiply substituted in any
position of the pyridine ring not already substituted by
1 0 -C-OI-I 9
: is selected from the group consisting of hydrogen, alkyl and substi-
tuted alkyl wherein the alkyl portion is from l to 8 carbon atoms,
inclusive, and isomeric forms thereof, cycloalkyl and substituted
cycloalkylg substituted oxygen, substituted nitrogen, halogen, phenyl
and substituted phenyl; -(CH2)~-OH, -(CH2)~-NR4R5, and isomeric
forms thereof, wherein m is an integer of from 1 to 8~ inclusive,
; R4 and R5 ar~ H or alkyl of from 1 to 8 carbon atoms, inclusive,
and isomeric forms thereof, wherein
' 11 i
-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-lin
;: aminide, 7(S)-hydroxy-methyl l-thlo-a-lincosaminide~ 7(5)-halo-
methyl l-thiO-a-linGosaminide, 7(R)-halo-methyl l-thiO-a-lincos-
~:~ aminide, 7(S)-methoxy-methyl l-thio-a-lincosaminide9 7-deoxy-7(S)-
(methylthio)-methyl l-thlo-a-lincosaminide~:7-deoxy-7(s)-(2-hydr
. ethylthio)-methyl l-thio-~-lincosaminide, and 7-deoxy-7(S)-(3-
hydroxypropylthio)-methyl l-thio--lincosaminide; there are obtained
: 30 novel and useful compounds of formula Io
Upon reacting an amino acid of the formula
CH
N
~ ) , 3
.
.
-8- . 3809A
//o
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,
C2H5, 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 react;ng an acid of the formulae
HO-C ~ R
E A
\ B ~
R1
HO-C// f
B
wherein A, B, D and ~ are selected from the group consisting of
nitrogen, oxygen~ sulfur and CR1R~; 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; -C-OH can be attached
to any ring carbon or nitrogen atom, with a sugar amine compound
selected from the group as defined abo~e, there are obtained novel
and useful compounds of formulae VI and VII.
MTL is methyl l-thio-~-lincosaminide of the formula
CH3
HO C-H
M~ H2N CH
~ 30 . H ~ ~
: ~ CH3
OH
~: 35
epi-MTL is methyl 7(S)-7-deoxy-7-hydroxy-1-thio--lincos-
aminide of the formula
.
-9- 3809A
CH3
H -C-OH
H2N--c,u
H V~--\l
~SCH3
OH
7-Cl-MTL is methyl 7(s)-7-deoxy-7-chloro-l-thio-a
lincosaminide of the formula
. fH3
: ~: 15 H-C-Cl
H2N~ ÇH
H 0~--O
2 0 ~ C H 3
O H
: ~ epi-7-Cl-MTL is methyl 7(R~-7-deoxy-7-chloro-1-thio
-l incosaminide of the formula
2 5
: ~:: ru
l - C - H
H a N ~-~ H
3 0~ ~ ~ H O/
~CH3
:`:: : OH
~: : 3 5 ~
7~S3-~ethoxyr-met~yl l rt~To-a-l ~ncosamTni~de ç~n ~e
:: ~ shown as follows (See U.S. 3,702,322, Example l, Part B-l):
.
:::
,
.' ~ , .
-1 0- 3809A
CH3
H-C -O-CH3
5 NH2-CH
H o~a~l
~S -CH3
OH
With reference to the following formula, 7-deoxy-7(S)-
(methyl thio) -methyl 1 -thi O-a-l i ncosami nide exi sts when R
is CH3; 7-deoxy-7(S)-(2-hydroxyethyl thio)-methyl l-thio-
a-lincosaminide exists when R is -CH2-CH2-OH; and, 7-
deoxy-7(S)-(3-hydroxypropylthio~-methyl l-thio-a-lincos-
amin;de exists when R is -CH2-CH2-CH2-OH (See l,'.S. 3,915,954, Examples
. ~: 1, 10 and 31):
C H 3
I
:: H-C-S-R
NH~-C H
H L/~ ~
~\H~
OH.
The hydroxy and halo groups at the 7 position of
the above formulas can be shown as ~ol lows
CH3
- H2N--CH
H V~
~CH3
OH
.
11- 3809A
wherein Y is selected from the group consisting of 7(R)
hydroxy, 7(S)-hydroxy, 7(S)- halo , and 7(R)- halo
~hen a pyridine acyl group is used, the result;ng
analog can ~e 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 isomersO Generally,
for any of the compounds described herein, the reduced
form is more antibacterially-active than the unsaturated
precursor. The use of a piperid;ne 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
more 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.
C'~xed Carboxylic Acid Anhydride Procedure," Chemistry of
The Amino Acids, Vol. 2, p. 970, John ~iley and Sons, Inc,
19610) When the acid is unsàturated, the resulting
unsaturated analog can be catalytically reduced under
standard conditîons to prepare the saturated analog, For
example, the reduction can be conducted using the follow-
. ing conditions
H2 at S to 50 psi
Catalyst - platinium oxide (Pt 2 )
Solvent - H20 or H20 ~ MeOH, or H20 + EtOH
HCl - 10~ excess
Time - 24 to 48 hours
As used herein~ a1k~1 o~ 1 t~ 8 can~on at~s~,
~nclusive, and isomeric forms thereof, includes methyl,
ethyl 9 propyl, butyl, pentyl, hexyl, heptyl, octyl, and
branched chain isomers thereof.
$ubstituted alkyl means the above a1kyl compounds in
which one or more of the hydrogen atoms has been replaced
by a halogen7 i.e., Cl, Br, F, and I, oxygen, hydroxylJ
amine ~primary)~ amine gsecondary-alkyl substituted by
alkyl as above3, amine (tertiary-alkyl substituted by
3L~l6. D3~
-12- 3809A
alkyl as above), sulfur, -SH, and phenyl. Exemplary
compounds are l-fluoroethyl, l-chloroethyl, 2-fluoroethyl,
2-chloroethyl, l-bromopropyl, 2~iodopropyl, l-chlorobutyl~
4-fluorobwtyl, and 4-chlorobutyl.
Cycloalkyl means 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, hydroxyt, amine (pr;mary, secondary9
and tertiary with the latter two alkyl substituted as
above), -SH, and phenyl. Exemplary compounds are p-
lS bromophenyl, m iodophenyl, o-chlorophenyl, p-ethylphenyl 9
m-propylphenyl, o-methylpheny1, and p-octylphenyl.
As det2iled infra, the compounds of the invention
can be phosphorylated to 9ive the 2-phosphate, and
acylated to give the 2-palrnitate which are both anti-
bacterially-active in vivo.
Substituted oxygen means oxygen substituted by an alkyl of
from 1 to 8 carbons, inclusive3 aryl, and substituted arylO
Substituted nitrogen means nitrogen substituted by an acyl of
from 2 to 18 carbons, a monoalkyl of 1-8 carbons, înclusive9 and a
dialkyl, wherein the alkyl is from 1 to 8 carbons, inclusive, includ
ing the iso~eric forms for all acyl and alkyl groups~
; Halo means chloro, bromo, iodo, or fluoro.
Exemplary sources for the amino acids used as starting
mRterials herein are as follows:
30 ~ 1. Heterocyclic Compounds7 Vol. 1, John Wiley and Sons~ Inc.9
1950. This sou~e describes the preparation of halogen
and alkyl substituted amino acids~
~; 2. Chem. Abstractso
81 - 105223A - alkyl and cycloalkyl
81 ~ 152243S - alkyl and halogen subst;tuted
8Z - 170746H - halogen substituted
85 - 46322Q - dihalo substituted
: :
' ' .
3~
-13- 3809A
8$ - 177258W - dihalo substituted
84 - 116928X - dihalo substituted
81 - 3737d - phenyl substituted
78 - 58201t - phenyl substituted
76 - 126800y - tetrahalo substituted
82 - 11036K - bromo substituted
83 27119~1 - bromo substituted
84 - 16613X - bromo substituted
78 - 123494G - bromo substituted
_ Br
1 0 CH2CH2-CH-CH3
\~
84 - 135488V ~
~N ~ OOH
.
CH~-O
: 81 - 151951J ~
Br ~ ~ ~ ~ COOH
B r - CH 2
8~ - i7809~
: . N `COOH
~ ~ ,
O CH3
:84 - 30918G - ~ ~
CQOH
C2H5-O O-C~H5
l 33139C - ~
~ ~ ~ ~ COOH
79 - 19109V - ~
COOH
:
~.
:. . ' . .
.
:
s~
-1 4- 3809A
81 - 3737D
5 . ~3~
~N COOH
O ~
8t - 13~964K - ~ ~ COOH
I H-C-CH3
85 - 1 77349B -~f OOH
~ _ OC 21~ 5
~COOH
7 8 - 71 8 6 5 G ~J i
2 0 C l ~ ~N
81 - 1 3 5 9 6 4 K ~ ~ C O OH
Ct O N
25~ COOH
$3 - 147397G - ~3\
CH
- :: 3 0 3
C H 3
82 ~ 11036K - ~NH 2
COOH
.
`
:, , ` ` , `: . .
6.'~
-l S- 3809A
~Çl
84 - 116928X - Cl~N COOh
C2Hs~O~ Q-C~Hs
81 - 3313gC ~
N OOH
Cl
Cl~ Cl
76 - 126800Y
Cl N~ COOH
~C2Hs
Cs~
: 79 - 115449b - ~ ;
~ , N COOH
: : 20
NH-CH3
Gl
~Cl
67 - 63229K
N COOH
: 25
CH3
: ~ - .
~ ~ ~ 68 - 104926b ~ ~ ~COOH
: ~
:~ 30
CH3~,~
69 - 59048Z
` N COOH
.
~;
'
.~. . .
6 ~
-16- 3809A
71 - 124907M H- ~ C-OH
S
~R
H-N~N C-OH
H
O
68 - 59465N 0 ~ COl-O-CH3
This compound can be hydrolyzed to the acid
by means well known in the art9 which acid
can then be reduced, also by means well
known in the artO
86 - 106501 e l/CH3
2 5 ~\ C _ N
H3
This compound can be hydrolyzed to the acid
~H ~ by means well known in the art. The result-
30 ~ ing acid then can be N-demethylated by the
u ~ . procedures disclosed in U.SO Patent 3,58399720
:
69 67282M CH I ~
CH3
:
, ,
, . ,
6 ~
_17_ 3809A
This compound can be hydrolyzed to the acid
by means well known in the art.
CH~
90 - 7029/-14-Z CH3~
CH3
This compound can be hydrolyzed to the acid
: 10 by means well known in the art. Also~ one
or both of the N-CH3 groups can be removed
from the resulting acid by following the
- procedures disclosed in U.S. Patent
3,583,972.
1 5 ~
9 - 168488X o ~ C-O-C4Hg
CH3-~-H
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
2~ ~ RC ~H
H
,
by methods disclosed in U.S. Patent
30 : ~ 3,583,9720
: 85 - 142995G~ NH2 ~ CH2 ~
~ 35 COOH
: ~ H
: ~
,.
~6~3~
~ 3809
81 - 15202~S CH3-CH-CH2-CH
OH ~ l
~ 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. Pr~tection of such groups is well known
in the art. See Protective Groups in Organic Chemistry,
J. F. W. M~Omie, Plenum Publishing Co., Ltd., 1973.
Jour. Chem. Soc. 1969 - '2134
Various H-alkyl substituted pyrid~nes
O
-N ~-R CHzR
2S ~ ~ R-MgX
commerciall~
available)
~ 30 R = alkyl, branched alkyl and cycloalkyl
:: :: :: :
4. Jour. Chem. Soc. 1969 - 934 -
~ NH, ~r ~ C_N ~ C-OH
~. ~
653
-19- 3809
The following examples are illustrative of the
process and products of the invention, but are not to be
construed as limiting. All percentages are by weight and
all solvent mixture proport~ons are by volume unless
otherwise noted.
Example l_- 4-Cis~et yl-~-pi Qeco?ic acid amide of 7-Cl-
MTL ' HCl (U-57,930E - Compound V~
PART I
C2H~ oHCl l2H~ CH3
7-Cl-MTL ~ ~3~ o
C-OH ~ NH ~H
HO~ O
~S-CH3
2 0 OH
A solution o~ 67 9 (0,357 moles) of the amino acid~HCl
(C.A. 51, 1643a, 1957) and 71.5 9 (0.714 moies) of
triethyla~ine dissolved in 2.5 liters of acetonitrile is
cooled to 10 C and 47.6 y (0.354 moles) of isobutyl-
chloroformate added in one portionO This mixture
(Solut;on A~ is stirred at 10 C for l hour. Solution
3 ;s made up by dissolYiny 97,7 9 ~0~357 moles) of 7-Cl-
MTL (J. Med. ChemO, 12-780, 1969, B, J. Magerlein`and F.
Kagan) in a warm mixture of 1500 ml of acetone and 1500
ml of H200 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 vacuumO The white~ mushy residue is fil~ered and
the crystalline material collected and dried to give 95 9
of pure productO Workup of the filtrate (chromatography~
5 3~.
20- 3809~
gave another 10 9 of product. The overall yield is 73%0
Anal. Calcd~ for C17H2sClN205S: C, 50.42; H, 6.22; N,
6.92; S, 7.92; Cl, 8.76.
. found: C~ 50.67; H~ 6.40; N, 6.64; S, 7,90; Cl,
8.7~.
~CHCl3 ~C~ 1.0) ~ 293
PART II
_
1 0
~2~5 C2H5 CH3
lS ~ -7-C1-MTL R C~H -CH XHzO
~Cl
~OH
~ / S-C1~3
Y bH
A mixture of 4.05 g (0.01 mole) of starting material,
40 ml of water., 60 ml of met~anolg 1.0 ml of 37% HCl
and 8.0 g of PTOa 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 l:l. 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
savedO The white s~lid3 which was the most polar of the
two products observed upon TLC of the reduction mixture,
3S was recrystallized from water to give the desired product,
U~57,930E~ m.p. 222-224, ln a yield of from 25 to 35X.
Anal~ Calcd. ~or C1,H3aCl2NzO5S: C, 45.63; H, 7.21; N,
~.~l6~3~
I
-21-.. 3809A
~.26; S, 7.17; Cl, 15.85.
Found: C, 45.77; H, 7.44; N3 6.39; S, 7.21; Cl,
16.17.
aH20 (C, 1.0) ~ 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 antimicrob-al spectra
Table I
The Minimal Inhibitory Concentra~ion of U-57,930
.: and Clindamycin Ys. Aerobic Bacteria.
:
~IC (~qJml)
:: Orqanism UC Cl;ndamycin U-57,930E
Staphyl ococcus 6685 >25 >25
: ::
dureus 6686 - . .05 .20
2 0
~ 6 ~ 7 . 0 2 5 . 2 0
66~ >25 >25
; ~ ~689 ~ OS . 78
6 h 9 0 . . 0 2 5 . 2 0
2 S : 6 6 9 1 . 1 ~ . 20
6 ~9 2 > 2 5 . . > 2 5
Ç693 .05 .78
6 6 9 4 > 2 5 ~ 2 5
66g5 . lO .~9
30~ : 6696 .10 .39
.
6 ~ 7 ~ . ~ 5 . 3 9
7 6 ~ 0 5 . 1 0
746 <.05 .05
57 1 . 20 . 78
3 5
'
5~
-22- 3809A
57~ .20 .39
Staphylococcus 719 .~0 .20
epidermid;s 3389 .10 .20
Streptococcus
~aecalis 694 25 6.25
Streptococcus
pyogenes 152 C.012 ~.012
Streptococcus
viridans 153 <.012 .05
87~ <.012 <.012 ,
Diplococcus
pneumoniae I- 41 <.012 <.012
Oi pl ococcus
zo pneumoniae II3Z13 <~,û12 <.012
.
Escheri chi a col i 45 50 >50
Proteus vulgaris93 >50 >50
25.
~1 eb-s i el l a
pneumoniae 58 6.Z5 >50
:: ~
Salmonella
schottmuelleri126 ~50 >50
' - ,. . .
............ ..... Pseudomonas
: aeruginosa 9S ~50 ~50
~ 35 The procedure for the above test is as ~ollows~
The Minimal Inhibitory Concentration (MIC's) of both
compounds Vs. aerobic bacteria is determined using a
3~
-23- 3809A
standard microplate broth dilution rnethod. Brain Heart
Infusion (BHI - Difco~ broth medium is used, and the
plates are ;ncubated at 37 C for 20 hrs.
: S. aureus UC 6685-6696 are clinical isolates which
5 are resistant to one or more co~mercial antibiotics.
"UC" is a registered trademark of The Upjohn Company
Culture Collection. These cultures can be obtained from
The Upjohn Company in Kalamazoo, M;chigan, upon request.
Table II
The Minimal Inhibitory Concentration of Cl;ndamycin and
U-57930E vs. Gram-Positive and Gram-NegatiYe Anaerobic
Bac~eria
MIC(~g/ml
Or~anism UC Clindamycin U-57930E
Bacteroides
fragilis 6513 0.06 C.12
6428 . 0 . 06 0 . 25
6864 3.9 2.0
6~62 7.~ 15,6
: 8acteroides
: : thetaiotaomicron 6512 200 û.5
,
~: Bacteroides
distasonis 6518 0.12 <0.03
~ 30
:~ Bacteroides
melaninogen_cus 6326 0.06 0.06
Clostridium
~ 247 0, 06 - 0 . 1 2
. 6509 0,06 0.12
Olostridium
.
no~yi B 6329 0.06 0.l2
: .
6~
- 2~ - 3809A
Clostridiurn
-
tert_um 6508 7.8 7.8
C l o st r i d i u m
-
cadaveris 6510 <0.03 0.06
Clostridium
sordel 1 i i 6505 2.0 0. S
Cl ostr_di um
0 t entan i 6521 <0 O 03 <0.03
C l o s t r d i u m
botulinum A 6506 0025 <0.03
l S Ct ost ri d i um
-
bifermentans 6507 0.50 0~06
Cl ostri d; um~
di~icile 6834 7.8 3.9
6857 250 125
6858 3.9 3.9
6860 500 50~
6861 3.9 200
25 Pro~onl bacteri um
acnes 6~64 0.06 0.12
6575 ~0.~3 0.06
Eubacterium
l imosum 6515 2.0 2.0
Eubacteri um
lentum 6522 0O50 1.0
~,s
naeslundii 5920 ~.25 0.25
-25- 3809A
Fusobacterium
nucleatum 6516 0.1~ 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 Oq2
Peptococcus
magnus 6258 0~06 0.06
Peptococcu~
aerogenes 6319 <0.03 0.05
I
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 o~ Schaedler 8roth~ and 9.0 ml of molten (47 C3
Wilkens Chalgren Agar Medium, in~ra,is added to the
antibiotic-supplemented brotL, After mixing with the
antibiotic, the agar is poured into lO0 mm x Z0 mm petri
. dishes. The dishes are allowed to stand on th-e bench
overnight prior to inoculationO
: Cultures are streaked on Wilkens-Chalgren Agar, and
: grown for 48 hours at 37 C in a 8BL Anaerobe Jar. Growth
:: From the plate is harvested, and a cell suspension is made
in Schaedler broth to equal the turbidity o~ a 0~
McFarland Standard ( lO8 cellsJml)~ The suspension is
pipetted into the wells of a Stee~s replicator, and
~pprox. 1-2 ~1 is deliyered to the sur~ace ~ the asar
-26- 3809A
plates. After allowing a few minutes for the inoculum to
dry, the plates are placed in a B8L Anaerobe Jar (atmos-
phere o~ 85% N, 10% H, 5% CO2) and incubated at 37 C for
72 hours.
S The Minimal Inhibitory Concentrat;on (MIC) is read
as the least amount of drug that inhibits growth. A very
faint film of growth, or ~3 colonies is considered
negative.
Wilkins-Chalg _n Agar Medium
Dispense the following ingredients and dissolve in
1000 ml disti1led wa~er. The pH should be 7.0 - 7.20
Trypticase 10 9
Gelysate 10 g
Yeast Extract 5 9
lS Elucose l g
NaCl . s g
L-Arginine-Free e ase l g
Py-ruvic Acid-Sodium Salt 1 g
Agar 15 g
Add Heme and ~Vitamin K1 solutions to yield
final concentrations of 5 ~g/ml Hemin and 0.5
g/ml Ki
AutoclaYe at 121 C For 15 minutes aerobicallyO
Heme Stock - 0.5 g Hemin ~ lO ml 1 N NaOH + 990
~l ~2
Autoclave at 121 C for 12 minutesO
Add lO ml stosk per liter of medium.
Yitamin K Stock - 005 ml Vit. K1 solution ~ 20
~ ~ ml 95% ethanol
- 30 Filter sterilize
Add 0.2 ml stock per liter of medium
The I.P~ ~Dso 0~ 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 determinationsO, This value
is approximately 2 times the LD50 for clindamycin HCl.
The LDso value should be interpreted as indicating that
the acute r.P. toxîcity of U-57,930E is approximately
one-half that of clindamycin HCl.
l~ÇiS~
-27- 3809A
In vivo . Mouse Protection Test
_ _ _
Or~anism & Route C1 indamycin U-57930A Ratio
S. aureus
_. _
Subcut.5.7(4.Z-7.8)* ~<5
Oral 12.3~8.8-17.3) 1-5 ca.10
_ hemolyticus
Subcut. 2.3(1.6-3.3) .25 ca.10
Subcut. 3.3(2.6-4.2) .25(0.2-0.33) 13
Oral 12~3(1002-14~8) 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-1 Mice~ weighing 18-20 gm were
infected with approximately 100 LD50'S of standardized
bacterial ce~ll suspensions w~ich had been maintained
frozen at -170~ C. Immediately before use, the suspen-
sions were thawed quickly and properly diluted. Infec-
t;on was via the intraperitoneal routeO
Treatment of the infected groups was begun immediate-
ly and continued once per day for 4 days (ti rst 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 anima1s were sacrificed and the median protect^
ive dose of the antibiotic calculated on the basis of
mor~ality ra~es i.n the treatmen~ groups. The ~edian
protectiYe dose and its 95~ confidence interval were
calculated according to ~he met~od of Spearmen & Karber
as programmed on a 360 digital computer.
Also isolated from Example I, Par~ II, is compound
V A. This material is obtained as follows.
The filtrate wh;ch was saved from Part II was con-
centrated to dryness under vacuum9 ~he residue converte~
to its free base and chromatographed over silica gel using
6t~
' -23- 3809A
CHCl3:methanol (6:1) as the eluting solvent. In this
manner the least polar material mentioned in Part II was
obtained. It was converted to its HCl salt and recrystal-
1 i zed from acetone and water. This isomer is tentatively
being assigned structure Y A.
C2H5
lo ~J4~ C//() NH ¦H
H HO ~ ~
V A ~ S-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 isolated by conventional procedures such ascrystallization or chromatography.
; l2Hs
V eDimerize~ ~ V B
N -7-Cl-MTL
Ç2Hs
e imeriZ ~
V A ~ ~ l Q
N ~ ~ ~ 7ycl-MT~
Alternatively, V and V A 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 F and V G may be coupled with any of
3 ~
-29 3809A
the lincosaminides described earlierO
C2Hs C2Hs
_
Hydrolize ~ e~i.. eri~ O
N 'C-OH N ~ -OH
V D H V F H
C2Hs C2H5
V Hydrolize ~ ~ epimerize ~ ~
~~ N " ~-OH
H H
VE V G
The D-cis isomer (V A) of U-57,930E has an anti-
bacterial spectrum when tested on BHl ~roth as described
~:: . previously in Table 1.
: Organism UC No. MIC (~g/ml)
S. aureus 76 250
570 100
746 125
: : S. fecalis 694 ~1000
S.~p~ogenes 152 62.5
:: D. pneumoniae 41 62.5
E. coli 45 ,looo
K. pneumoniae 58 ~1000
S schottmuelleri lZ6 ~1000
: Ps. aeruqinosa 95 >1000
~:
Example 2 Other Analogs of 7-Cl~MTLo
By following the procedures of Example 1, but
substituting the amino acid with the following amino
acids there are prepared the corresponding novel
~653~
, . 30_ 3809A
antibacterially-active analogs as their free bases or
acid addition salts. The latter can be prepared by
methods well-known to ~hose skilled in the art.
Amino Acid Analo~
~J-45,863
N ~L C - OH
C-OH
U-46,138
N
: HO-C=O
1S L
: ~ U-46,137
~J '''
N
O U-46,337
~ ~ C-OH ~Fast isomer on TLC-
: N MeOH:CHCl 95:5. Run on
silica gel plates~
U-46,~65
N ~ C OH (Slow isomer on TLC~
H
30 : :
~ E: U-46,69~
C-OH (Slow isomer on TLC,
prepared from U-46,465)
CH3
3s
:~
-31- 3809A
HsC2 -~L8 U-45 ,656
~ C-OH
N
C2Hg
~ ~ 8 U-45,652
N C-OH
C2H5~_ ll U-46 ,701
N C~OH
CH3
C2H~LII U-60,481
C-OH
N
. H
U-44,469
2 5 N - O H
C H 2
IC H 2
O H .
3 0 O
~C-OH
3 5
tiS3~;
-32- 3809A
Example 3 - Analogs 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. Bannister and H. Hoeksema) ~or 7-
C1-MTL, there are prepared the corresponding novel anti-
bacterially-active analogs:
Ami no Ac;id Anal_ogs
U-46, 136
[~L R
G-OH
C;~Hs
C-OH U-45,653
U-60,493 (fast
isomer TLC)
O
2 5 H ~-OH
~ U-~0,492 ~slow
~ i somer TLC~
N C-OH
H
Example_4 - Analogs~ Q~ MT~
By following the procedures of Example 1, ~ut
substituting the amino acid with the following amino
acids, and substituting epi-MTL (J. Chem. Soc, Perkin I
~.-~
33 3809A
1974, p. 360-8, 8annister) for 7-Cl-~T~ there are prepared
the correspond;ng novel antibacterially-act;v.e analogs:
Ami no. dC i d hnaloq
~ o U-46,135
F2H~ . U-.45,6~9 -
C-OH
: . .
Cpd. A
~: '~ O
~ N ~ -OH
H
C2Hs
, ~ Cpd. 8.
: ~ N
H
~ ~ .
:
Ex~mple 5 -. Analo~s o~ epT~.7-.Cl..~lT~
By~foltowing thé procedures- of Exa~pte 1, but
:: substituting:the am;no;ac;d with the ~ollowing aminQ
acids, and substituting epi-7-Cl=MTL for 7-Cl-MTL, there
are prepared the correspond.ing novel antibacteriall~-
-active analogs~ .
~; ~S
~ .
3~L~
-34- 3809A
Ami n o A c i d A n a l o ~
~LII Cpd~ C
~N C - O H
C 2Hs
:~ 10 ~ 1I Cpd. D
, ~N
Cpd, E
N
H
2~5
Cpd~ F
-OH
N
Z5
Epi-7-Cl~MTL can be prepared b~ the procedure used
to prepare 7-C1-MTL with t~e exceptlon that the starting
: ma~eria:l is epi:-~TL instead o~ MTL~
~: 30 Chèmical and ph~stcal c~aracterization o~ most of
the compounds o~ Examples 2~5 are as ~ollows~ -
.
- 3 5 - 38 O9A
C H N S C l a D . ~I p .
,
U-45,863 1 47.80 5.62 7.44 8.51 9.41 CHC13
2 ~7.76 5.54 7.35 8.76 9.34 +286~ 96-100
U-46,138 1 47.80 5.62 7.~4 8.51 9041
~ 47.30 5.74 6091 8.51 9.44
.
10U-46,137 1 47.80 5.62 7.44 8.51 9.41 EtOH
2 47058 5.74 7.56 8.53 9~49 -~216 189-190
U-46,136 1 50.25 6.19 7.82 8.95 MeOH
2 49.16 5.86 8.01 9.15 +194 199-201
U-46,135 1 50.26 6.19 7.82 8.95 MeOH
2 50.5~ ~.19 7.98 9.27 ~269 97-100
.
U-46,337A 1 42.96 6.73 6.68 7.65 16.91 EtOH
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
U-46,699E 1 44.34 6.98 6.47 7.40 16.36 H20
2 44.79 7024 6.25 7.36 16.~2 +1~2 229-234
U-45,656 1 50.~2 6.2~ 6.92 7.92 8~76 C~C13
2 50.85 6.39 6.7~ 7.54 ~3.S3 +250
.
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
: : ~
45,653 1 52.83 6.78 7.25 8.30 - MeOH
2 53.83 7.~8 7.39 8.14 ~ ~203
U-45,659 1 5Z.83 6.78 7~?5 8.30 - CHC13
2 52.77 6.70 7.34 8.55 - ~295
i5~
-3~- 3809A
U-46, 701 A 146 . 85 7 . 43 6 . 07 6 . 9S 15 . 37
2 ~
U-44,469E 144.06 6.96 6.05 6.92 15.30
24~.~3 ~ .68 6 .07 6 .72 1 5 .49
U-45,6S7 143.80 4090 6.81 7.80 17.24 MeOH
243.5~ ~.93 6.82 7.82 17.41 fl81 105-130
10 1 ~ CALCD.
2 - FOUND
Example 6 - Fusaric Actd Amide of 7-Chloro-MTL.
H~3C ~
C - OH
CH3
HgC4i~3 HC-Cl
C--NH ~H
~ \~
2~ ~/S-CH3 ~u-55,
: 0~3
: .
: ~ By following the procedure oF Example 1, ~ut su~tir
tuting the amino acid with fusaric acid, there is
3 0o bta i n ed U- 5 5 , 581 ,
Anal. Calcd. for ~19H29C1Nz05S 0, 52~70i ~1, 6.75; N,
6.479 S, ~,41; Cl ~ 8.~9.
Found: C7 52.15; H, 6.65; N, 6~36; S, 7,~1; Cl ?
7 .94 .
Example 7 - 4-Cis-n-Butyl-L-Pipecolic Acid Amide of
7-Cl -MTL or U-60 ,970E
;
6 5~
7 3809A
CL,Hg C4H9
C-7-Cl-~l. HC//ONHHClHcl (X)H,O
H ~ Oy
I\g~
¦ I S-CH3
OH
(Cj7H32Cl2N205S (X)H20)
A mixture of 4.0 g (0.0093 mole) of starting material,
40 ml of water, 40 ml of methanol, 2 ml'of 37% HCl, and
lS 8.0 9. PtO2 catalyst were reduced on a Parr hydrogenator
at 50 psi for 18 hours. The reaction was filtered to
remove the catalyst and the filtrate concentrated under
vacuum to give an amber oil. The oil was dissolved in
20 ml. o~ a 2:1 solution of CHCl 3 and methanol and
enough triethylamine added to neutralize the HCl pre-
sentO This solution was then chromatographed over silica
gel using a so'lvent system composed of CHCl3:methanol
(2:1~. Two main product fractions are obtained. The
fract'ions containing the faster moving material were
pooled and evaporated under vacuum to give a white solidg
fract;on Ao ~he 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 37% 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 Cl7H32Cl2N205S: C, 47.99; H, 7.63; N,
5.89; S, 6.75; Cl, 14.920
Found: C, 47.97; H, 7.42, N, 6.23; S, 6.90; Cl,
14.870
6 ~
-38- 3809A
MeOH + 178 (C, 1.0)
CMR analysis supports the proposed structure.
S The Minimal Inhibitory Concentration (MIC) in ~g/ml
of U-60,970E against various bacteria is as follows:
Organism UC# MIC
S. aureus 76 0.125
. 570 0.25
746 0.062
S. faecal;s 694 0.25
S. pyogenes 152 0.008
__
D pneumoniae 41 0.016
_ coli 45 31.2
lS K. pneumoniae 58 7.8
S. schottmuelleri126 31.2
: Ps. aeruginosa 95 >125
: The test procedure is as disclosed in Example 1.
~ U-60,970E was also tested in vivo in standard lab-
: ~ 20 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,g70E
: is significantly ~ore active in vivo against D. pneumoniae
n ~ 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~
~ ~ ~ 35
: ~ ~
: ~
L 6 ~
39 3809A
._ _ ,
_ ,~ ,_ ~ .
~o r~ c~l ~ 03
. a~ ~o ~ In
I I I I
I L~
rr~ L~
S_ r~ ~D ~ C~J C~J
V o ~
oo '`l ~ ~ '`'
.--._ ,_ ~
X C~J co ~ ~ d-
a~ ~
_ O ~O ~ .
V l V~ . . . .
~, ~ ~ ._ ~ o o
U~
s~: u~ O u~ ~ C`J
.' ~ ~ .- o o o
~ o_~
C V~ . ooo
~n _ _ ~
~ 15 o~,_ ~
~-- d- r~ CO
~ ~ ,_ ~
~ C~
Lf~
' l ~ CO .~, , -
d~
o . ~_ r r~ o . o
~L
~_ o ~_ _
~, .- C~ d
a
: ~ ~ ~ CO
~o
C~ ~ ,~
~_ ~-- cn ,,-- ,_ _~,
~7 L~
U~j C~ r,~
o I I
a) o~ ~ c~ oo I
~:: 2 5 _ c~ ,~ . co ~D
o o .~
_. _, _ ,_
oo r_ ~ cO
o Ln ~Q
:: ~ U) o . ~ ; -
_ _ . .:.
I_
` ~: 30 .- _ ,_
~5 ~ ,~
: . o ,_
. ,_ ~
` 3 5 F ~ el C
. _ o ~ ~ ~
- ~ Q a~
~, _v~ '" o C~I
,
~ ,,
''
-4Q- 3809A
Example 8 - l-D-Pipecolic Acid Amide of
7-Cl-_TL or U-61,734E
C~Hg ,C4Hg
~ C-7-Cl-MTL ~ ~C-NH-CH
H
H ~)~
S-CH3
OH
(C17H32ClN205S (X)H20~
Fraction A from the preceding experiment was converted to its HCl
salt in the same manner as described for fraction ~. A
25-35% yield of product was obtained whose CMR spectrum
: was essentially identical to that obtained from fraction
B.
Example 9 - Preparation of a Compound in ~hich the Amino-
acid Portion Contains a Heteroatom In a 5-
: Membered _ing
¦ ! H~drplysis~
CH 3~ CH3-~
CH 3 ~H3
+
7-C1-MTL
CH3
; C~3-~ C// HC-~l
\ NH - CH
CH3 H~ ~ O
:~ ~ SCH3
OH
:-
-41- 3809A
The aminoacid ester (see C.A. 69 - 67282M) 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 or the zwitterion. The coupling of the aminoacid
HCl with 7-Cl-MTL is accomplished in the same manner as
described in Example 19 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 - Preearation o~ a Compound in Which the Amino-
acid Portion Contains a Heteroatom in a 6-
Membered Ring
~O ~ ~ ~O
`N C-O-CH3 N C-OH
Cl Cl
CH3 +
CH3 ~ HC-Cl 7-Cl- ~L
~N ~ NH - CH
~-CH I
OH
Cl
The aminoacid ester (see C.A. 6~ - 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
-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 g. (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 Analogs
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
ts those skilled ;n-the art--which 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 1-lOcan be prepared by procedures well-known ~o
those skilled in the artO By obvious appropriate modifi~
cation9 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 arepresentative num~er- of the compounds prepared in
Examples 2-8 follows. The test procedure is as given
supra.
.
6~3~L~
43 _ 3809A
~ o~o
L5'~ N L O O O O O ~`1
~ ~t - C`J ~I O O O O C~ O l ~ .
1~ ~ A ~ ~ A A . ~2:
l _ _ _ _ _ ~ _ _ _ _ _ _
LC~ n u~ 3 ~ o g o oO g o
~ ~C ~ ~ ~ O O O O O O O
~ ~ ~ A A ~ ~
r- ~ _ __ _ _ _ _ _ _ _ __ _
O ,1 ~ ~ u~ ~n o o o o o o
I~ ~ ~C ~ C~l C~ l O O O O O O
~ Z- ~ --~D l~ O O O O O
e 1- ~ ~ --r~ A A A O =
5 _ _ _ _ _ _ _ _ _ _ _ _ _ _
.--~ L~ o o o o o o o o o o ol o
E~u~ /~ o o o o o o o o o o ol O
, o o o L~ o o o o o o olL~
~ ~ Lr~ _, ,_ .--~ .-- ~ .--.--~ .,
E~ ~ ~ . A ~ ~ J~
.CI a ' ' - - - - - - - - - - - ~
vl 5 hn ~l ol ol o o o o ol o o ol o
~ ol l o ~ O .o¦ .--roo--gi =~
~¦~r ~ ¦ A ¦ A A A A A I A A A I 1~
-I - I 1- --------I _ T
ILC~ O; ol ol o ~1 - o ol o ol ol o
1~1 ~ ol gl ol n ol ¦ o¦ ~c~ ol l ~3
IL~ ~ _1 ~1 I ~_~ ~ ~
~ ~ ¦' I I ! ~--~ ! -- I ~
1~ x ~ I ~ I I 1-
~L~ ~ CJI~ ol o olo olo
ol oo ol Q ~1 ol o=~
I I I --I I ~ I i
:: ~ ~1 ~ T ~ ~1~
I I ~ 1'~
, ol ~, ,, , ~, ~, , ~,
~1 r l Cl l I Vl O I ~I aJI I
1~~1 ~1 ~ o l 1 ~1 a l
~: I ~ ~ ol
¦ ~ I o ~ i
1 l a ~a l E l ~1 ~ I E
5 1 5 1 ~ 1 E I ~ I ~> 1 ~ I Ql I ~
~1 ~1 ~1 a~l~1 ~ I ::~1 ~I L>
"~ I a ~ ~ 1 ' I I ~ I > ~ o
'~ ~ ~. ~
.
.
.
6s~
44 3809A
..
~ X ~:n co cs: o7 ~ _ _ _~--O ~____
o- I ~ r~ r_ ~ u~ o o o o o ~ ~ aJ
~D ~Z~ --O O 1~7 A O ~ L
_ ,,_'_ _ _ _ _ _ _ _ _ _ _ _ : 2~ 0
L~ X ~ ~ a o cl~ o s_~
~ ~ ~ ~) V ~ . O O L" O O V 3 E-
~ ' _ _ _ _ _ ~ _ _ _ ~ _ ~--o
I~x e~
__l O G O O O O O O O O O Q~ _
~) ~ o o o ~ o o o o o o L~ ~ E ~--
D O O ,_ C`J O O O O ----N ~n O
LO 1`1 _ __ _ _ _ _ _ _ A _ L~ O
~ ~ o o 8 oO o o o g o o g
D ~ O O O O - O O O O O ~
a~ __ _ A _ _ _ _ _ _ _ _ _
~ ~D ~
- ~ ~ _~ O O O O O O O O O O O
E s_ ~ <~ ~oo- o o o oo-- ,o --o roo--o o
E ~ ~ ~. ~ A _ ~ _ ~ ~ ~ ~ ~ ~ O_t~
~ ~ ~ O O O O C:~ O O O O O O ll
E ,_ _~ o o A O A . O O O O O O J .
X N ¦ ¦ N ¦ o l ¦ o l o l O o l L~ O ~
co ~ ~O I I lol lololo olo ~o ll
~ c~ --I ! I_A~ 1~--X
~1 l
I ~ =~ ~rl
1. I Vl ~O~ l l ~1 ~--1 V
' l ~1 '1 ~1 1 1 'l '~ ~1 ' '~1
I~ u~ 1 1 1 ~)1 ~ a
t~ ~ I s~ Q~l 0 ~1
=~ ~ ~ ~ I
L~l ~ 81 1 ~I s. l ~1 'O ol
=~ ~ ,1 C ol s ~l
s ~1 5-l El ,ul I ~1 ~1 ~u cl ~ a~l
~I :~1 ~1 ~1 1 l =)l ~ 5~ Cl
~ ~1 :al ~1 1 1 ~1 >I o u~l 'Y
v~ ~
~ t3 ~
3809A
MIC ln mcq/ml
U-Number and Structure
. U-60,493 U-60,492U-60,481
_ . , _ _ . ,
S aureus UC 76 1000 1000 2.0
__ __ _ ~ . . _ ,
S. aureus UC 5701000 ~1000 3.9
. _ ~
10S~ aureus UC 746 250 250 2.Q
S. h~nolyticus UC 152 7.8 15.6 >1.0
._, _
St. faecalis UC 694 _ lQOO 1000 _ 31.2 ,_ _ .
S. lutea UC 130
~ . .
E. coli UC 45 >1000 >1000 500
P. vulgaris UC 93 >1000 >1000 1000
K. pneumon;ae UC S8 >1000 ~1000 250
. ~ ~
S. Schottmuelleri UC 126 >1000 >1000 1000
_ _ _ ~
Ps, aeruginosa UC g~ ~1000 ~1000 ~1000
O ~ M~ 15.6 31.2
~ ~ast Slow
isomer - isomer
on TLC on TLC IR &
. NMR OK
O O
11 11
X=C-7-01 ~MTL' - Y-C-MTL
. R=C2H5
. ~
Since the compounds of the subject invention are
active agai nst Yari OU5 Gram-positive and Gram-negative
microbes, they can be used ;n various environments to
inhibit such microbes- For example, they can be used as
d;sinfectants to inhibit S- aureus on washed and stacked
~ood utensils contaminated with this bacterium. They alsn
can be used as disinfectants on various dental and medioal
equipment contaminated with S. aureus. Further9 the
compounds of the inYention can be used as bacteriostatic
rinses for laundered clothes, and for impregnating papers
~ 6 3809A
and fabrics; 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 ~orms according to the pH of
the environ~ent. ~hen 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 conver~ed to stable acid~addition saits by
neutrali2ing t~e free ~ase with the appropriate acid,
about pH 7.0, and advan~ageousl~ to about p~ 2 to pH 6.
Suitable acids for this purpose include hydrochloric,
sulfuric, phosphoric, thiocyanic, fluosilicic, hexa~luoro-
arsenic, hexafluorophosp~oric, acetic, succin;c~ citric,lactic, maleicl fumaric, pamoic, cholic, palmi~ic, mucic,
camphoric, glutaric, glycolic, phthalic, tartaric? lauric,
stearic, salicylic, 3-phenylsalicylic, 5-phenylsalicylic,
3-methylglutaric, orthosulfobenzoic~ cyclohexanesulfamic,
ZO cyclopentanepropionic, 1,2-crclohexanedicarboxylicl 4-
cyclohexanecarboxylic, oc~adecen~lsucc~nic, octenyl-
succinic, methanesulfonic, helianthic, Reinecke's7
dimethyldithiocarbamic, hexadecylsulramic~ octadecyl-
sulfamic, sorbic, monochloroacetic, undecylenic, 4'-
hydroxyazobenzene-~-sulfonic, octadecylsul~uric, picric,
benzoic, cinnamic, and like acids.
The acid-addition salts can be used for the same
purposes as the free base or the~ 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,
wh;ch can be subjected to purif;cat;on procedures, ~or
example, solvent extractions and washings7 chromatography9
fraction~l liquid-liquid extractions, and crystallization9
and then used to regenerate the free base form by treat-
ment with alkali or to make a different salt by meta-
thes;sO Or the free base can ~e converted to a water-
soluble salt~ such as the hydrochloride or sulfate, and
~ ~6~3.~
~ ~ -47- 3809A
the aqueous solution o~ the salt extracted with various
water-immiscible solvents before regenerating the free
base iorm by treatment of the thus-extracted acid solu-
tion, or converted to another salt by meta-thesis.
S 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~ ~ormaldehyde forms resinous materia1s useful as
pickliny inhibitors according to U.S, Patent Nos.
2,425,320 and 2,606,155. T~e free bases also ma~e good
Yehicles for toxic acids. For example, t~e Fluosilicic
acid addit;on salts are useful as mothproofing agents
according to U.S~ patent Nos~ 1,915,334 and 2?075~359 and
the hexafluoroarsenic acid and hexafluorophosphori-c acid
addition salts are useful as parasiticides according to
U.S. Patent Nos. 3,122,536 and 3,122,55~,
The compounds of the su~ject invention are useful as
ant;bacterial- a~ents in suita~le compositions, These
compositions are preferabl~ presented for administration
to humans and animals in unit dosage forms, such as
tablets, capsules ? pi lls, po~ders, granules, sterile
parenteral solutions or suspensions, and ~ral solutions
or suspensions, a~d oil-water emulsions containing suit-
a~le quantities of the active compound in t~e ~orm of the
Free base? or its pharmacologicall~ accepta61e salts.
For oral administration? either solid or fluid unit
dosa~e ~orms can ~e prepared, For preparing soljd compo-
sitions such as ta~lets, the principal active ingredient
is mixed with conventional ingredients such as talc,
3Q magnesium stearate~ dicalcium phosphate, magnesium
aluminum silicate, calcium sul~ate, starch, lactose,
acacia, methylcellulose, and functionally similar mater-
;als as pharmaceutical diluents or carriers. The tablets
can be laminated or otherwise compounded to provide a
dosage form afiording the advantage o~ prolonged or
delayed action or predetermined successive action of
the enclosed medication. For example, the tab7et can
~ ~.53~i
4~ 3809A
comprise an inner dosage and an outer dosa~e 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 ~he inner component to pass intact
into the duodenum or to be delayed in release~ A variety
of materials can be used for such enteric layers or coat-
ings, such materials including a number of polymeric acids
or mixture o~ 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 incompati~le active
ingredients. Wafers are prepared in the same manner as
lS tablets, differing 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
20 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 slurry o~ 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, The
water-soluble forms of the active compound can be dis-
solved in an aqueous vehicle together with sugar,aromatic ~laYoring agents and preservatives to form a
syrup. An elixir is prepared by using a h~dro-alcoholic
(ethanol3 vehicle with suita~le sweeteners such as sucrose
- together with an aromatic flavoring agent. Suspensions
can be prepared of the insoluble ~orms with a syrup
vehicle with the aid of a suspending agent such as acacia,
tragacanth, methylcellulose and the lîke,
' -49- 38~9A
Topical ointments can be prepared by dispersing the
active compaund in a suitable ointment base such as
petrolatum, lanolin, polyethylene glyco1s, 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 ~ase. Topical creams and lotions are pre-
pared b~ dispersing the compound in the oil phase prior
to the emulsification of the oil phase in waterO
10 For parenteral administration, fluid unit dosage
- forms are prepared utilizing the active compound and a
sterile vehicle9 water being preferred. The active com-
pound~ depending on the form and concentration used, can
be either suspended or dissolved in the ~ehicle. In pre-
paring solutions, a water-soluble form o-F t~e active
compound can be dissolved in water for injection and
filter steril.ized ~efore filling into a suitable vial or
ampul and sealing.0 Advantageousl~ adjuvants such as a
local anesthetic9 preservative and ~uffering agents can
2~ be dissolved in the vehicle. To enhance the sta~ility,
the composition can 5e frozen after filling into the Yial
and the water rem.oved under vacuum. T~e dry lyophilized
powder is then sea.led in the vial and an accompanying vial
o~ 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 ~eing
dissolved and sterilization cannot be accomplished by
- filtra.tionO The active compound can be sterilized by
exposure to ethylene oxide before suspending the sterile
vehicle. Advantageously, a surfactant or wetting agent
is inciuded in the compos;tion to facilitate uniform
: distribut~on of the active compound.
The term unit dosage form as used in t~e specifica-
tion and claims re~ers to physicall~ disc.rete units suit-
able as .unitary dosages for human subjects and animals,
each unit containing a predetermined quantit~ of active
53~
-50~ 3809A
material calculated ~o produce the desired therapeutic
e~fect in association with the required pharmaceutical
diluent, carrier or vehicle. The spec;fications for the
novel unit dosage forms of this invention are dictated
by and directly dependent on (a) the uni.que characteris-
tics of the active material and the particular thera-
peutic effect to be acnieved, 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 this specification, these ~eing features of the
present invention. Examples oF suitable unit dosage forms
in accord with this invention are ta~lets, capsules, pills,
troches, supposito.ries, powder packets, granules, wafers,
cachets, teaspoonfuls, ta~lespoonfuls, dropper-Fuls, ampu1s,
vials, segregated.multiples of any cf t~e foregoing, and
other forms as herein descri~ed,
In addition to the administration of the active comY
pound as the princ.ipal active ;ngred~ent of compositions
for the treatment o~ the conditions described ~erei.n, the
said compound can ~e included with other types of com~
pounds to obtain ad~antageous comGinations of properties~
Such combina.tions i.nclude the acti~e compound with anti-
biotics such as spectinomycin, chloramphenicol, novo-
~iocin, dih~dronovobioc.in, tetracyclines (eeg., tetra-
cycline, oxytetracycline and c~lortetracy~cline)? peni-
cillin~,. erythromyci:n ? kanamycin, streptom~cin ? neomycin ?
~ polymyx;n, ~acltractn~ nystatin, filipin, fuma~illin ~nd
: :: endom~cin to Broaden the bacterial spectrum of the compo-
sition and for s~nergistic action against particular
bacteria.;. steroids having anti-inflammatory actiyity such
as h~drocorti~one~ predn~solone, 6~-meth~lpredn;solone,
6~-fluoroprednisolone and the like; analgesics such as
aspirin, sodium salic~late ~ace~lsalic~lic acid3
: anh~dride, N-acetyl-p-aminophenyl and salicylamide;
antihistaminesg suc~ as chlorpheniramine ~aieate,
diphenylhydramine, promethaz;ne, p~rathiazine, and the
like, sulfas, such as sulfadlazine, sul~amethazine~
a.
-51- 3~09A
sulfamerazine sulface-tam~de, 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 dail~ (ever~ six hours), embraces the
ef~ective range for the treatment o~ most conditions for
which the compositions are e~fective, for ch~ldren, the
dosage is calculated on the basis o~ 15 to 30 mg,/kg./day
to be administered every six hours.
The active compound is compounded with a suitable
pharmaceutical carrier in unit dosage form for convenient
and effectiYe 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 65% w/v for
parenteral treatment. The dosage of compositions contain
ing the active compound and one or more other active
ingredients is to he determined with reference to the
usual dosage o~ each such ingredient.
The following examples are illustrative of the best
mode contemplated by the inventor for carrying out his
in~ention and are not to be construed as limitingO
The examples use U-57,930E or U-60,970E as the active compound,
but it-should be-understood that this is only exemplary --
o~ the other active compounds of the subject invention.
As distinguished from the prev;ous examples, the follow-
ing are Composition Examples O
Composition Example 1
Capsules
3S One thousand two-piece hard gelatin capsules for oral
use, each containing 250 mg. of U-57,930E or U-60,970E are prepared
from the following types and amounts of materials:
~6~
-52- 3809A
Gm.
U-57,930E or U-60,970E 250
Corn starch lOO
Talc 7S .
Magnesium stearate 25
The materials are thoroughly mixed and then encap-
sulated in the usual manner.
The foregoing capsules are useful for the systemic
treatment of infection in adult humans by the oral
10 adminisl:ration of 1 capsule every 4 hours.
Using the procedure a~ove, capsules are similarly
prepared containing U-57,930E or U-60,970E in 15, 30, 50, 125 and 500
mg. amounts by substituting 15, 303 50, 125~ and 500 gm
of U-57,930E or U-60,970E for the 250 gm used aboYe.
15 Composition Example 2
Capsules
One thousand two~piece ~ard gelati.n 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 o~ ingredients~ .
: : G~,
U-57,930E or U~60,970E 200
Tetracycline ~drochloride 25
Talc 75
llagnesium stearate 25
:: The ingredients are thoroughl~ mixed and then encap-
: sulated in the usual manner~
The foregoing capsules are useful for khe systemic
~: ~ treatment of infection in adult humans b~ the oral
~: ~ 3~ administration o~ 1 capsule every~ 6 hours,
Using :the procedure above3 capsules are similarly
prepared~containing U-57,930E or U-60,970E and each of the following
antibiotics -i-n p~ace- ~f- tet~.ac.~.cl.ine..~y su~stituting 250
: ~ gm. o~ such other antibiotic for tetracycline: chlor-
: . 35 amphenicol, oxytetrac~cline, chlortetracycline, fumagillin9
erythromycin, streptomycin, dihydronovobiocin and novo
biocinO When a penic~llin, such as potassi.um penicillin
~,
. .
, . .
5~
-53-
3809A
G, is to be used in place of tetracycline, 250,000 units
per capsule is employed.
Such combinat;on products are useful for the system-
ic treatment of mixed infections in adult humans by the
S oral administration of 1 capsule every 6 hours.
Composition Example 3
Tablets
One thousand tablets for oral use, each containing
500 mg. of U-57,930E or U 60,970E are prePared from the following
types and amounts of materials:
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.
Z5 Using the above procedure, except for reducing the
- ' amount of U-57,930E or U-60,970E to 250 gm., tablets containing 250
mg. o~ U-57,930E or U-60,970E are prepared.
Composition Exampl Q4
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
sulfadiazine, sul~amerazine, and sulfamethazine, are pre-
pared from ~he fol'lowi'ng types'~a~nd''amounts of materials
~m.
U-57,930E or U-60~970E 250
Sul fadiazine 83.3
Sulfamerazine 83.3
" ~ 5 3~i
-5~- 3809A
Sulfamethazine 83.3
Lactose 50
Corn starch SO
Calcium stearate 25
S Light liqu;d petrolatum 5
The intredients are thoroughly mixed and slugged.
The slubs are broken down by forcing through a number
sixteen screen. The resulting granules are then com-
pressed into tablets, each containing 250 mg. of U-$7,930E
or U-60,970E and a total of 2S0 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 hoursO
For the treatment of urinary infections, the triple
sulfas in the above formulation is advantageously
replaced by 250 gm. of sulfamethylthiadia.zole or 250 gm.
of sulfaceta~ide.
Composition Example 5
ZO 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. o~ U-57,930E or U-60,970E is prepared from the
following types and amounts of ingredients:
Gm.
U-57,930Eor U-60,~70E 50
Sulfadiazine 33.3
Sulfamerazine 3303
Sulfamethazine 33.3
Citric acid 2
Benzoic acid
Sucrose 700
Tragacanth 5
Lemon oil 2 ccO
Deionized water, q.s. 1,000 ccO
The citric acid, benzoîc acid, sucrose, tragacanth,
and lemon oil are dispersed in sufficient water to make
-55- 3809A
850 cc. of solution. The U-57,930E or U-60,970E and finely powdered
sulfas are stirred into the syrup until uniformly dis-
tributed. Sufficient water is added to make 1,000 cc.
The composition so prepared is useful in the systemic
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 from
the following types and amounts of materials:
Gm.
U-57,930E or U-60,970E 200
Lidocaine hydrochloride 4
Methylparaben 2.5
Water for injection, q.s. 1,000 cc.
The ingredients are dissolved in water and the solu-
tion sterilized by filtration. The sterile solution is
filled into vials and the vials sealed.
Z0 Composition Example 7
Parenteral Pre~aration
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 ~rom the following
types and amounts of ingredients:
Gm O
U-57,930E or U-60~970~ 200
Spectinomycin sulfate 400
Lactose
- 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 sterili~ed by
filtration. The sterile solution, in the amount of 2 cc.,
is aseptically f;lled into sterile vials and~ frozen. The
water is removed under high vacuum and the vials contain-
ing the lyophilized powder are sealedO Just prior ~o use,
sufficient sterile water for injection to make 2 cc. of
6 ~ 3 ~
6- 3809A
solution is added to the vial.
Composition Exa~le 8
Topical Ointment
One thousand gm. of 0025% ointment is prepared from
5 the ~ollowing types and amounts o~ ingredients:
Gm.
U-57,930E or U-60,970E 2.5
Zinc oxide 50
Calam;ne 50
Liquid petrolatum (heavy) 250
Wool fa~ 200
White petrolatumg q.sO 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 ~he powders are
finely divided and uniformly dispersed. The powder mix-
ture is stirred into the white petrolatum mixture and
stirring continued until the ointment congealsO
ZO The ~oregoing 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 calamineO
Following the procedure above, ointments are similarly
prepared cont~ining U-57,930E or U-60,970E in 0.5, 1, 2, and 5X
amounts by substituting 5, 109 20 and 50 gm. o~ U-57,930E or
: U-60,:970E for the 2.5 gm. used above.
C~position Example 9
Cream
One thousand gm. of a vaginal cream are prepared from
the following types and amoun~s o~ ingredients:
Gm O
U-57,93~E or U~60,970~ 50
Tegacid Regular1 1~0
Spermaceti 100
Propylene glycol 50
Polysorba'e 0 5
:
' :
-57- 3809A
Methylparaben
Deionized water, q.s. 1,000 gm.
Self-emulsifying glyceryl monostearate from
Goldschmidt Chemical Corporation, New York, N.Y.
The Tegacid and spermacetl are melted together at a
temperature of 70-80 C. The methylparaben is dissolved
in about 500 gm. o~ ~ater and the propylene glycol,
Polysorbate ~0, 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 minutes with continued stirring until the
temperature has dropped to 40-45 C. The pH of the final
cream is adjusted to 3.5 by incorporating 2.5 gm. of
citric acid and 0.2 g. of dibasic sodium phosphate dis-
solved in about 50 gm. of water. Finally, suffic;ent
water is added to bring the final weight to 1,000 gm. and
the preparation stirred to maintain homogeneity until
cooled and congealedO
The foregoing composition is useful for the treat~
ment of vaginal infections in humansO
Composition Exam~le 10
Ointment, Ophthalmic
One thousand gm. of an ophthalmic ointment containing
0.5~ U-57,930E or U-60,970E are prepared from the following types and
amounts of ingredientso
GmO
U-57,930E or U~60,970~ 5
Bacitracin 12.2
Polymyxin B sul~ate tlO,OOO
units/mg.)
Light liquid petrolatum 250
Wool fat 200
White petrolatum, q.s. 1,000 gmO
The solid ingredients are finely divided by means of
an air micronizer and added to the light liquid petrolatum.
The mixture is passed through a colloid mill to uniformly
3~
~5~~ 3~09A
distribute the micronized particles. The wool fat and
white petrolatum are melted toyether, 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
5 gm. (0.5~) of methylprednisolone for the treatment o~
inflammation, and, alternatively, the bacitracin and
polymyxin B sulfate can be omitted
Composition_Example 11
Eye-Ear Drops
One thousand cc. of a sterile aqueous solution for
eye or ear use containing 10 mg. ~f U-57,930E or U-60~970E and 5 mg.
of methylprednisolone in each cc. is prepared from the
following types and amounts of ingredients:
GmO
U-57,930~ or U~60,970E 10
Methylprednisolone phosphate sodium S
Sodium citrate 4.5
Sodium bisulfite
Polyethylene glycol 4000 120
Myristyl-y-picoliniu~ chloride 0.2
Polyvinylpyrrolidone
Deionized water, q.s. ad 1000 cc~
~ The in~redients are dissolved in the water and the
resulting solution is steril;zed by filtration. The
solution is aseptically filled into sterile dropper
containersO
The composition so prepared is useful in the ~opical
treatment of inflammation and infection of the eye and ear
as well as o~her sensitive tissues of the animal body.
Composition Example 12
Troches
tj3~6
-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 sulfate 50
Polymyxin B sulfate (10,000
units/mg.~ 1
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 of humans.
Composition Example_13
~ Suppository, Rectal
~ One thousand suppositories, each weighing 2.5 gm. and
20 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
unitslm~.) 1.25
; Methylprednisolone
Ethyl aminobenzoate 75
; Zinc oxide 62.5
Propylene glycol 162.5
` 30 ~ Polyethylene glycol 4,000 q.sO
2,500 gm.
;The U-579930E or U-60,970E, polymyxin B sulfate, methylpredniso-
lone, ethyl aminobenzoate3 and zinc oxide are added to
the propylene glycol and the mixture milled until the
pow~ers are finely diYided and uniformly dispersed. The
polyethylene glycol 4000 is melted and the propylene
glycol dispersion added slowly with stirring. The
. , ~ .
.
6 ~
' -60- 3809A
suspension is poured into unchilled molds at 40 C.
The composit;on is 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.
Compos;tion Examele 14
Mastitis 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.
U-57,930E or U-60,970E 25
Methylprednisolone acetate 0.5
Light liquid petrolatum 300
Chlorobutanol, anhydrous 5
Polysorbate 80 5
2Q - 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 light liquid petrolatum until finely
25 divided and uniformly dispersed. The chlorobutanol,
s 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-
30 ture and is filled into disposable mastitis syringes in
10 gm. doses.
Composition Example 15
Animal Feed
One thousand gm. of a feed mix is prepared from the
35 following types and amounts of ingredients:
Gm O
U-57,930 or U-60,970E 10
-61- 3809A
Soybean meal 400
Fish meal 400
Wheat germ oil 50
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 pigs, 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
g;ve the desired dose of U-57,930Eor U-60,970E.
Composit~on 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 p~ovide therapeutic propertiesO
Similarly, each of the above free base compounds can
be used in the form o~ 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
o~ the above antibacterially-active invention compounds
can be substituted as the active ingredient to provide
compositlons having therapeutic properties.
.
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