CA1164864A - Analogs of lincomycin and clindamycin - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
Novel useful analogs of the well known antibiotics lincomycin and clindamycin. These analogs are prepared by condensing a cyclic acid with a sugar amine and are re!presented by the general formula:

wherein A, B and E are nitrogen, CH3-N?, oxygen, sulfur or CR1R1;
wherein R1, 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, cyclo-alkyl and substituted cycloalkyl, substituted oxygen, sub-stituted nitrogen, halogen, phenyl and substituted phenyl or -(CH2)m-OH, -(CH2)m-NR4R5, and isomeric 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 there-of; and wherein R2, which can be attached to any ring carbon, not already substituted by Rl, or nitrogen atom, is:
-?-X

and X is the amino function of 7(R)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(R)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(S)-methoxy-methyl 1-thio-.alpha.-linco-saminide, 7-deoxy-7(S)-(methylthio)-methyl 1-thio-.alpha.-linco-saminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl 1-thio-.alpha.-lincosaminide or 7-deoxy-7(S)-(3-hydroxypropylthio)-methyl 1-thio-.alpha.-lincosaminide.

Description

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 ¦ 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 ma~/

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 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 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

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:

~ H2)n ~ R

J III
N

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, 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~

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 ~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 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 ~

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

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 HO-C-H
HzN - CH
H ~ ~

OH
epi-MTL is methyl 7(S)-7-deoxy-7-hydroxy-l-thio-~-lincos-aminide of the formula 1 16~86~

H-C-OH

\~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 Cl -C-H

~
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~

H ~ ~

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):

H-C--S-R
NHz-CH
Hp ~ ~

OH.

The hydroxy and halo groups at the 7 position of the above formulas can be shown as follows ~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~

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~

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 ~N "-`COOH

81 - 151951J - ~ lI
~ Br~`~ ~ COOH

Br-CH
81 - i7809a - I ~
N COOH

84 - 30918G - ~
COOH

C2Hs-O ~ o-C2Hs 81 - 33139C - l ll ..~N'~ COOH

I
79 - l9109Y -. ~`N ~ ~COOH

1 16486~

~N ~ COOH

81 - 135964K - ~ ~ COOH

85 - 177349B - ~ OOH

OC2Hs ~ COOH

Cl ~ ~N ~

81 - 135964K - Cl ~ o ~ COOH

COOH

83 - 147397G - \
-- ~N Cl 82 ~ 11036K - ~ NH2 ~ COOH

1 16486~

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 N COOH

68 - 1 04926b ~3~

6 9 - 5 9 04 8 Z ~S~
COOH

1 16486~

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

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 -17- ` 3809A
This compound can be hydrolyzed to the acid by means well known in the art.

90 - 7029/-14-Z 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

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

~ 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

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~

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

~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 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~

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 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 -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 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.

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 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.

- 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.

.. ~

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 the lineosaminides described earlier.

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

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 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) - 1 16486~

Hsca ~8 U-45,656 ~N C-OH

U-45,652 C-OH

C2Hs~L~ U-46,701 -OH
N

C2Hs~L_o U- 6 0, 481 C-OH
2 0 . H

~ U-44,469 . ~IN~-OH

OH

C - OH
~ ~L Cl U-45,657 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

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 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 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 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 ' -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 ~

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 ~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.

_ _ .~ .
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 ~ ~

.

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

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

+

7-Cl -MTL

CH3~ ~ ~ NH--CH

CH3 H ~ 0~

~CH3 OH

- ` 11648B4 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

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~

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.

~ ~ ~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 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 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

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 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 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 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 .
-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 16~8~4 850 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 -~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 -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 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 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.

Claims (4)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A process for preparing a compound of the general formula:

wherein A, B and E are selected from the group consisting of nitrogen, and CR1R1; wherein R1, which can be singly or multiply attached to any ring carbon atom not already substituted by R2, is selected from the group consisting of hydrogen, alkyl of from 1 to 8 carbon atoms, inclusive, and isomeric forms thereof; and wherein R2, which can be attached to any ring carbon, not already substituted by R1, or nitrogen atom, is:

-?-X

and X is the amino function of a compound selected from the group consisting of 7(R)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(R)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(S)-methoxy-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl 1-thio-a-lincosaminide and 7-deoxy-7(S)-(3-hydroxypropylthio)-methyl 1-thio-.alpha.-lincosaminide;
which comprises reacting an acid of the general formula:

wherein A, B, E and R1 are as defined above, and the -COOH group can be attached to any ring carbon, not already substituted by R1, or nitrogen atom, with a sugar amine compound selected from the group consisting of 7(R)-hydroxy-methyl 1-thio-.alpha.-lincos-aminide, 7(S)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(R)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(S)-methoxy-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl 1-thio-.alpha.-lincosaminide and 7-deoxy-7(S)-(3-hydroxypropylthio)-methyl 1-thio-.alpha.-lincosaminide;
and, when required, preparing a pharmaceutically acceptable acid-addition salt.

2. A process for preparing a compound of the formula:

which comprises reacting an acid of the formula:

with a compound of the formula:

and, when required, preparing a pharmaceutically acceptable acid-addition salt.

3. A compound of the general formula:

wherein A, B and E are selected from the group consisting of nitrogen, and CR1R1; wherein R1, which can be singly or multiply attached to any ring carbon atom not already substituted by R2, is selected from the group consisting of hydrogen, alkyl of from 1 to 8 carbon atoms, inclusive, and isomeric forms thereof; and wherein R2, which can be attached to any ring carbon, not already suhstituted by R1, or nitrogen atom, is:

-?-X

and X is the amino function of a compound selected from the group consisting of 7(R)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-hydroxy-methyl 1-thio-.alpha.-lincosaminide, 7(S)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(R)-halo-methyl 1-thio-.alpha.-lincosaminide, 7(S)-methoxy-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl 1-thio-.alpha.-lincosaminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl 1-thio-.alpha.-lincosaminide and 7-deoxy-7(S)-(3-hydroxypropylthio)-methyl 1-thio-.alpha.-lincosaminide;

and the pharmaceutically acceptable acid-addition salts thereof;
when produced by the process defined in claim 1 or an obvious chemical equivalent thereof.

4. A compound of the formula:

and the pharmaceutically acceptable acid-addition salts thereof;
when produced by the process defined in claim 2 or an obvious chemical equivalent thereof.