U.S. patent application number 12/163299 was filed with the patent office on 2008-11-20 for novel lipopeptides as antibacterial agents.
Invention is credited to Dale Christensen, John Finn, Jason Hill, Dennis Keith, Tsvetelina Lazarova, Michael Morytko, Ian Parr, Jim Siedlecki, Jared Silverman, Alan D. Watson, Xiang Yang Yu, Yan Zhang.
Application Number | 20080287347 12/163299 |
Document ID | / |
Family ID | 26866577 |
Filed Date | 2008-11-20 |
United States Patent
Application |
20080287347 |
Kind Code |
A1 |
Hill; Jason ; et
al. |
November 20, 2008 |
Novel Lipopeptides as Antibacterial Agents
Abstract
The present invention relates to novel lipopeptide compounds.
The invention also relates to pharmaceutical compositions of these
compounds and methods of using these compounds as antibacterial
compounds. The invention also relates to methods of producing these
novel lipopeptide compounds and intermediates used in producing
these compounds.
Inventors: |
Hill; Jason; (Auburndale,
MA) ; Parr; Ian; (Medford, MA) ; Morytko;
Michael; (Framingham, MA) ; Siedlecki; Jim;
(Burlington, MA) ; Yu; Xiang Yang; (Billerica,
MA) ; Silverman; Jared; (Brookline, MA) ;
Keith; Dennis; (Arlington, MA) ; Finn; John;
(Stow, MA) ; Christensen; Dale; (Apex, NC)
; Lazarova; Tsvetelina; (Brookline, MA) ; Watson;
Alan D.; (Lexington, MA) ; Zhang; Yan;
(Sharon, MA) |
Correspondence
Address: |
Intellectual Property Department;Cubist Pharmaceuticals, Inc.
65 Hayden Avenue
Lexington
MA
02421
US
|
Family ID: |
26866577 |
Appl. No.: |
12/163299 |
Filed: |
June 27, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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|
09737908 |
Dec 15, 2000 |
7408025 |
|
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12163299 |
|
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60170946 |
Dec 15, 1999 |
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60208222 |
May 30, 2000 |
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Current U.S.
Class: |
514/1.1 ;
530/317 |
Current CPC
Class: |
C07K 7/08 20130101; A61K
38/00 20130101; A61P 31/04 20180101 |
Class at
Publication: |
514/11 ;
530/317 |
International
Class: |
A61K 38/12 20060101
A61K038/12; C07K 7/54 20060101 C07K007/54; A61P 31/04 20060101
A61P031/04 |
Claims
1. A compound having the formula (I): ##STR01497## and salts
thereof, wherein R is: ##STR01498## wherein X and X'' are
independently C.dbd.O, C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X, S.dbd.O
or SO.sub.2; wherein n is 0 or 1; wherein R.sup.X is alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl,
hydroxyl, alkoxy, carboxy or carboalkoxy; wherein B is X''R.sup.Y,
H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or
heterocyclyl; wherein R.sup.Y is hydrido, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl or hydroxyl; wherein A
is H, NH.sub.2, NHR.sup.A, NR.sup.AR.sup.B, alkyl, alkenyl,
alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or
heterocyclyl; wherein R.sup.A and R.sup.B are independently alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
carboalkoxy; wherein when n is 0, then A is additionally:
##STR01499## wherein each of R.sup.50-R.sup.53 is independently
C.sub.1-C.sub.15 alkyl; alternatively, wherein B and A together
form a 5-7 membered heterocyclic or heteroaryl ring; wherein
R.sup.1 is ##STR01500## wherein X' and X''' are independently
C.dbd.O, C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X', S.dbd.O or SO.sub.2;
wherein m is 0 or 1; wherein R.sup.X' is alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,
carboxy or carboalkoxy; wherein B' is X'''R.sup.Y', H, alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl;
wherein R.sup.Y' is hydrido, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, cycloalkyl, heterocyclyl or hydroxyl; wherein A' is H,
NH.sub.2, NHR.sup.A', NR.sup.A'R.sup.B', heteroaryl, cycloalkyl or
heterocyclyl; wherein R.sup.A' and R.sup.B' are independently
alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl
or carboalkoxy; wherein when m is 0, then A' is additionally:
##STR01501## wherein each of R.sup.50-R.sup.53 is independently
C.sub.1-C.sub.15 alkyl; provided that when B' is H and X' is
C.dbd.O, then A' is other than (a) a pyridinyl ring substituted
with one substitutent NHC(O)R.sup.D or (b) a C.sub.5-C.sub.6
saturated cycloalkyl ring substituted with one substitutent
NHC(O)R.sup.D; wherein R.sup.D is C.sub.1-C.sub.17 unsubstituted
alkyl or C.sub.2-C.sub.17 unsubstituted alkenyl; and when B' is H
and m=0, then A' is not H; wherein R.sup.2 is ##STR01502## wherein
K and K' together form a C.sub.3-C.sub.7 cycloalkyl or heterocyclyl
ring or a C.sub.5-C.sub.10 aryl or heteroaryl ring; wherein J is
hydrido, amino, NHR.sup.J, NR.sub.JR.sup.K, alkyl, alkenyl,
alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl,
heterocyclyl, alkylamino, hydroxyl, thio, alkylthio, alkenylthio,
sulfinyl, sulfonyl, azido, cyano, halo, ##STR01503## wherein each
of R.sup.24, R.sup.25, and R.sup.26 is independently alkyl,
cycloalkyl, heterocyclyl, aryl and heteroaryl; or R.sup.24 and
R.sup.25 together form a 5-8 membered heterocyclyl ring; wherein
R.sup.J and R.sup.K are independently alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl or heterocyclyl; or alternatively,
wherein J, together with R.sup.17, forms a 5-8 membered
heterocyclyl or cycloalkyl ring; or alternatively, wherein J,
together with both R.sup.17 and R.sup.18, forms a 5-8 membered
aryl, cycloalkyl, heterocyclyl or heteroaryl ring; and wherein each
of R.sup.17 and R.sup.18 is independently hydrido, halo, hydroxyl,
alkoxy, amino, thio, sulfinyl, sulfonyl or ##STR01504## or wherein
R.sup.17 and R.sup.18 taken together can form a ketal, thioketal,
##STR01505## wherein each of R.sup.22 and R.sup.23 is independently
hydrido or alkyl.
2. A compound having the formula (I): ##STR01506## and salts
thereof; wherein R is: ##STR01507## wherein X and X'' are
independently C.dbd.O, C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X, S.dbd.O
or SO.sub.2; wherein n is 0 or 1; wherein R.sup.X is alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl,
hydroxyl, alkoxy, carboxy or carboalkoxy; wherein B is X''R.sup.Y,
H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or
heterocyclyl; wherein R.sup.Y is hydrido, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl or hydroxyl; wherein A
is H, NH.sub.2, NHR.sup.A, NR.sup.AR.sup.B, alkyl, alkenyl,
alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or
heterocyclyl; wherein R.sup.A and R.sup.B are independently alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
carboalkoxy; wherein when n is 0, then A is additionally selected
from: ##STR01508## wherein each of R.sup.50-R.sup.53 is
independently C.sub.1-C.sub.15 alkyl; alternatively, wherein B and
A together form a 5-7 membered heterocyclic or heteroaryl ring;
wherein R.sup.1 is ##STR01509## wherein X' and X''' are
independently C.dbd.O, C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X', S.dbd.O
or SO.sub.2; wherein m is 0 or 1; wherein R.sup.X' is alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl,
hydroxyl, alkoxy, carboxy or carboalkoxy; wherein B' is
X'''R.sup.Y', H, alkyl, alkenyl, alkynyl, aryl, heteroaryl,
cycloalkyl or heterocyclyl; wherein R.sup.Y' is hydrido, alkyl,
alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
hydroxyl; wherein A' is aryl; provided that when B' is H and X' is
C.dbd.O, then A' is other than a phenyl ring substituted with
substitutent NHC(O)R.sup.D, wherein R.sup.D is C.sub.1-C.sub.17
unsubstituted alkyl or C.sub.2-C.sub.17 unsubstituted alkenyl,
wherein said phenyl ring may be further optionally substituted with
1-2 substituents independently selected from amino, nitro,
C.sub.1-C.sub.3 alkyl, hydroxyl, C.sub.1-C.sub.3 alkoxy, halo,
mercapto, C.sub.1-C.sub.3 alkylthio, carbamyl or C.sub.1-C.sub.3
alkyl carbamyl; wherein R.sup.2 is ##STR01510## wherein K and K'
together form a C.sub.3-C.sub.7 cycloalkyl or heterocyclyl ring or
a C.sub.5-C.sub.10 aryl or heteroaryl ring; wherein J is hydrido,
amino, NHR.sup.J, NR.sup.JR.sup.K, alkyl, alkenyl, alkynyl, alkoxy,
aryloxy, aryl, heteroaryl, cycloalkyl, heterocyclyl, alkylamino,
hydroxyl, thio, alkylthio, alkenylthio, sulfinyl, sulfonyl, azido,
cyano, halo, ##STR01511## wherein each of R.sup.24, R.sup.25, and
R.sup.26 is independently alkyl, cycloalkyl, heterocyclyl, aryl or
heteroaryl; or R.sup.24 and R.sup.25 together form a 5-8 membered
heterocyclyl ring; wherein R.sup.J and R.sup.K are independently
selected from alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl
or heterocyclyl; or alternatively, wherein J, together with
R.sup.17, forms a 5-8 membered heterocyclyl or cycloalkyl ring; or
alternatively, wherein J, together with both R.sup.17 and R.sup.18,
forms a 5-8 membered aryl, cycloalkyl, heterocyclyl or heteroaryl
ring; and wherein each of R.sup.17 and R.sup.18 is independently
hydrido, halo, hydroxyl, alkoxy, amino, thio, sulfinyl, sulfonyl or
##STR01512## or wherein R.sup.17 and R.sup.18 taken together can
form a ketal, thioketal, ##STR01513## wherein each of R.sup.22 and
R.sup.23 is independently hydrido or alkyl.
3. The compound according to either of claims 1 or 2, wherein R is
##STR01514## wherein each of R.sup.3, R.sup.4, R.sup.5, and R.sup.6
is independently hydrido, alkyl, aryl, heterocyclyl or heteroaryl,
and wherein R.sup.44 is s alkyl, aryl, heterocyclyl or
heteroaryl.
4. The compound according to claim 3, wherein R is ##STR01515##
wherein R.sup.4' is s alkyl, aryl-substituted alkyl, substituted
phenyl, heteroaryl, heterocyclyl, optionally substituted
(C.sub.8-C.sub.14)-straight chain alkyl or ##STR01516## wherein
R.sup.7 is an alkyl group.
5. The compound according to claim 4, wherein R is ##STR01517##
wherein X.sup.3 is chloro or trifluoromethyl and wherein q is 0 or
1.
6. The compound according to either of claims 1 or 2, wherein
R.sup.1 is ##STR01518## wherein each of R.sup.10 and R.sup.11 is
independently hydrido, alkyl, aryl, heterocyclyl or heteroaryl; and
wherein R.sup.13 is aryl.
7. The compound according to claim 6, wherein R.sup.1 is
##STR01519## wherein each of R.sup.10 and R.sup.11 is independently
hydrido or alkyl; and wherein X.sup.4 is fluoro or
trifluoromethyl.
8. The compound according to either of claims 1 or 2, wherein J is
hydrido, amino, azido or ##STR01520## wherein R.sup.17 and R.sup.18
taken together form a group selected from ketal, ##STR01521## or
wherein R.sup.17 is hydroxyl when R.sup.18 is hydrido; or wherein
J, together with R.sup.17, forms a heterocyclyl ring.
9. The compound according to claim 8, wherein R.sup.2 is
##STR01522## wherein R.sup.17 and R.sup.18 taken together form
##STR01523## wherein R.sup.22 is H or alkyl; and wherein R.sup.19
is hydrido, amino, azido or ##STR01524##
10. The compound according to claim 9, wherein R.sup.2 is
##STR01525##
11. The compound according to either of claims 1 or 2 wherein said
compound is TABLE-US-00005 Cpd # R R.sup.1 R.sup.2 1
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01526## ##STR01527## 2
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01528## ##STR01529## 3
NHCO(CH.sub.2).sub.8CH.sub.3 NHSO.sub.2Ph ##STR01530## 4
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01531## ##STR01532## 5
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01533## ##STR01534## 6
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01535## ##STR01536## 7
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01537## ##STR01538## 8
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01539## ##STR01540## 9
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01541## ##STR01542## 10
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01543## ##STR01544## 11
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01545## ##STR01546## 12
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01547## ##STR01548## 13
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01549## ##STR01550## 14
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01551## ##STR01552## 15
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01553## ##STR01554## 16
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01555## ##STR01556## 17
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01557## ##STR01558## 18
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01559## ##STR01560## 19
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01561## ##STR01562## 20
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01563## ##STR01564## 21
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01565## ##STR01566## 22
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01567## ##STR01568## 23
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01569## ##STR01570## 24
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01571## ##STR01572## 25
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01573## ##STR01574## 26
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01575## ##STR01576## 27
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01577## ##STR01578## 28
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01579## ##STR01580## 29
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01581## ##STR01582## 30
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01583## ##STR01584## 31
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01585## ##STR01586## 32
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01587## ##STR01588## 33
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01589## ##STR01590## 34
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01591## ##STR01592## 35
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01593## ##STR01594## 36
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01595## ##STR01596## 57
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01597## ##STR01598## 70
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01599## ##STR01600## 182
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01601## ##STR01602## 190
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01603## ##STR01604## 193
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01605## ##STR01606## 221
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01607## ##STR01608## 254
NHCO(CH.sub.2).sub.7CH.sub.3 ##STR01609## ##STR01610## 255
NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01611## ##STR01612## 256
NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01613## ##STR01614## 257
NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01615## ##STR01616## 258
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01617## ##STR01618## 259
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01619## ##STR01620## 260
NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01621## ##STR01622## 261
NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01623## ##STR01624## 262
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01625## ##STR01626## 263
##STR01627## ##STR01628## ##STR01629## 270
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01630## ##STR01631## 292
##STR01632## ##STR01633## ##STR01634## 293
NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01635## ##STR01636## 294
NHCO(CH.sub.2).sub.7CH.sub.3 ##STR01637## ##STR01638## 312
NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01639## ##STR01640## 313
NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01641## ##STR01642## 314
NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01643## ##STR01644## 342
NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01645## ##STR01646## 343
NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01647## ##STR01648## 344
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01649## ##STR01650## 350
##STR01651## ##STR01652## ##STR01653## 351
NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01654## ##STR01655## 352
NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01656## ##STR01657##
12. The compound according to claim 11 wherein the compound is
TABLE-US-00006 Cpd # R R.sup.1 R.sup.2 260
NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01658## ##STR01659## 262
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01660## ##STR01661## 344
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01662## ##STR01663##
13. A compound of formula (I) according to either of claim 1 or
claim 2, wherein R is NHCO--[(C.sub.6-C.sub.14)-alkyl]-CH.sub.3,
and R.sup.1 and R.sup.2 are selected from: TABLE-US-00007 R.sup.1
R.sup.2 ##STR01664## ##STR01665## ##STR01666## ##STR01667##
NHSO.sub.2Ph ##STR01668## ##STR01669## ##STR01670## ##STR01671##
##STR01672## ##STR01673## ##STR01674## ##STR01675## ##STR01676##
##STR01677## ##STR01678## ##STR01679## ##STR01680## ##STR01681##
##STR01682## ##STR01683## ##STR01684## ##STR01685## ##STR01686##
##STR01687## ##STR01688## ##STR01689## ##STR01690## ##STR01691##
##STR01692## ##STR01693## ##STR01694## ##STR01695## ##STR01696##
##STR01697## ##STR01698## ##STR01699## ##STR01700## ##STR01701##
##STR01702## ##STR01703## ##STR01704## ##STR01705## ##STR01706##
##STR01707## ##STR01708## ##STR01709## ##STR01710## ##STR01711##
##STR01712## ##STR01713## ##STR01714## ##STR01715## ##STR01716##
##STR01717## ##STR01718## ##STR01719## ##STR01720## ##STR01721##
##STR01722## ##STR01723## ##STR01724## ##STR01725## ##STR01726##
##STR01727## ##STR01728## ##STR01729## ##STR01730## ##STR01731##
##STR01732## ##STR01733## ##STR01734## ##STR01735## ##STR01736##
##STR01737## ##STR01738## ##STR01739## ##STR01740## ##STR01741##
##STR01742## ##STR01743## ##STR01744## ##STR01745## ##STR01746##
##STR01747## ##STR01748## ##STR01749## ##STR01750##
14. The compound according to claim 13, wherein R is
NHCO--[(CH.sub.2).sub.6-14]-CH.sub.3.
15. A pharmaceutical composition comprising the compound according
to either of claims 1 or 2 and a pharmaceutically acceptable
carrier.
16. A method of treating a bacterial infection in a subject,
comprising the step of administering the pharmaceutical composition
according to claim 15 to a subject in need thereof for a time and
under conditions effective to ameliorate said bacterial
infection.
17. The method according to claim 16, wherein said subject is a
human, an animal, a cell culture or a plant.
18. The method according to claim 16, wherein said bacterial
infection is caused by a gram-positive bacteria.
19. The method according to claim 18, wherein said bacteria is an
antibiotic-resistant bacteria.
20. The method according to claim 19, wherein said
antibiotic-resistant bacteria are resistant to vancomycin,
methicillin, glycopeptide antibiotics, penicillin or
daptomycin.
21. The method according to claim 16, further comprising the step
of co-administering more than one compound of Formula (I) according
to either of claims 1 or 2 to a subject in need thereof.
22. The method according to claim 16, further comprising the step
of co-administering a second antimicrobial agent wherein said
second antimicrobial agent is not included within the scope of
Formula (I).
23. The method according to claim 22, wherein said antimicrobial
agent is penicillins, carbapenems, cephalosporins, aminoglycosides,
bacitracin, gramicidin, mupirocin, chloramphenicol, thiamphenicol,
fusidate sodium, lincomycin, clindamycin, macrolides, novobiocin,
polymyxins, rifamycins, spectinomycin, tetracyclines, vancomycin,
teicoplanin, streptogramins, anti-folate agents, trimethoprim,
pyrimethamine, nitroimidazoles, quinolones, fluoroquinolones,
isoniazid, ethambutol, pyrazinamide, para-aminosalicylic acid
(PAS), cycloserine, capreomycin, ethionamide, prothionamide,
thiacetazone, viomycin, everninomicin, glycopeptide, glycylcycline,
ketolides, oxazolidinones, imipenen, amikacin, netilmicin,
fosfomycin, gentamicin, ceftriaxone, ZIRACIN
(56-deacetyl-57-demethyl-45-O-de(2-methyl-1-oxopropyl)-12-O-(2,3,6-trideo-
xy-3-C-methyl-4-O-methyl-3-nitro-alpha-L-arabino-hexopyranosyl)flambamycin-
), LY333328 (oritavancin), linezolid
(N-[[(5S)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxo-5-oxazolidinyl]methy-
l]acetamide), SYNERCID (dalfopristin-quinupristin), aztreonam
(2-[[(Z)-[1-(2-amino-4-thiazolyl)-2-[[(2S,3S)-2-methyl-4-oxo-1-sulfo-3-az-
etidinyl]amino]-2-oxoethylidene]amino]oxy]-2-methyl-propanoic
acid), metronidazole (2-methyl-5-nitro-1H-imidazole-1-ethanol),
epiroprim
(5-[[3,5-diethoxy-4-(1H-pyrrol-1-yl)phenyl]methyl]-2,4-pyrimidinediamine)-
, OCA-983
(1-[[(2S)-2-amino-3-methyl-1-oxobutyl]amino]-2,5-anhydro-3-S-[(4-
R,5S,6S)-2-carboxy-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2-
.0]hept-2-en-3-yl]-1,4-dideoxy-3-thio-D-threo-pentitol), GV-143253
(trinem),
sanfetrinem((1S,5S,8aS,8bR)-1,2,5,6,7,8,8a,8b-octahydro-1-[(1R)-
-1-hydroxyethyl]-5-methoxy-2-oxo-azeto[2,1-a]isoindole-4-carboxylic
acid), CS-834
((4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-3-[[(3R)-5-oxo--
3-pyrrolidinyl]thio]-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic
acid (2,2-dimethyl-1-oxopropoxy)methyl ester),
biapenem(6-[[(4R,5S,6S)-2-carboxy-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo--
1-azabicyclo[3.2.0]hept-2-en-3-yl]thio]-6,7-dihydro-5H-pyrazolo[1,2-a][1,2-
,4]triazol-4-ium inner salt), KA 159 (stipiamide), dynemicin A
((1S,4R,4aR,14S,14aS,18Z)-1,4,7,12,13,14-hexahydro-6,8,11-trihydroxy-3-me-
thoxy-1-methyl-7,12-dioxo-4a,14a-epoxy-4,14-[3]hexene[1,5]diynonaphtho[2,3-
-c]phenanthridine-2-carboxylic acid), DX8739
((4R,5S,6S)-3-[[(3S,5S)-5-[[4-[(2S)-5-amino-2-hydroxy-1-oxopentyl]-1-pipe-
razinyl]carbonyl]-3-pyrrolidinyl]thio]-6-[(1R)-1-hydroxyethyl]-4-methyl-7--
oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid), DU 6681
((4R,5S,6S)-3-[[(6S)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl]thio]-6-[-
(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carbox-
ylic acid), cefluprenam
((2E)-N-(2-amino-2-oxoethyl)-3-[(6R,7R)-7-[[(2Z)-(5-amino-1,2,4-thiadiazo-
l-3-yl)[(fluoro
methoxy)imino]acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-
-2-en-3-yl]-N-ethyl-N-methyl-2-propen-1-aminium inner salt), ER
35786
((4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-3-[[(3S,5S)-5-[(R)-hydroxy(3R)-3-pyrr-
olidinylmethyl]-3-pyrrolidinyl]thio]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hep-
t-2-ene-2-carboxylic acid monohydrochloride), cefoselis
((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxy
imino)acetyl]amino]-3-[[2,3-dihydro-2-(2-hydroxyethyl)-3-imino-1H-pyrazol-
-1-yl]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic
acid), sanfetrinem celexetil
((1S,5S,8aS,8bR)-1,2,5,6,7,8,8a,8b-octahydro-1-[(1R)-1-hydroxyethyl]-5-me-
thoxy-2-oxo-azeto[2,1-a]isoindole-4-carboxylic acid
1-[(cyclohexyloxy)carbonyl]oxy]ethyl ester), cefpirome
(1-[[(6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-2--
carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-6,7-dihydro--
5H-cyclopenta[b]pyridinium inner salt), HMR-3647
(3-de[(2,6-dideoxy-3-C-methyl-3-O-methyl-alpha-L-ribo-hexopyranosyl)oxy]--
11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl[[4-[4-(3-pyridinyl)-1H-i-
midazol-1-yl]butyl]imino]]-erythromycin), RU-59863 (C-7 catechol
substituted cephalosporin), KP 736
((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)[[(1,4-dihydro-1,5-dihydroxy-4-oxo-
-2-pyridinyl)methoxy]imino]acetyl]amino]-8-oxo-3-[(1,2,3-thiadiazol-5-ylth-
io)methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
disodium salt), Rifalazil
(1',4-didehydro-1-deoxy-1,4-dihydro-3'-hydroxy-5'-[4-(2-methylpropyl)-1-p-
iperazinyl]-1-oxo-rifamycin VIII, MEN 10700
((5R,6S)-3-[[(2-amino-2-oxoethyl)methylamino]methyl]-6-[(1R)-1-hydroxyeth-
yl]-7-oxo-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid),
lenapenem
((4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-3-[[(3S,5S)-5-[(1R)-1-hydro-
xy-3-(methylamino)propyl]-3-pyrrolidinyl]thio]-4-methyl-7-oxo-1-azabicyclo-
[3.2.0]hept-2-ene-2-carboxylic acid), BO 2502A
((4R,5S,6S)-3-[(2S,3'S,4S)-[2,3'-bipyrrolidin]-4-ylthio]-6-[(1R)-1-hydrox-
yethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic
acid), NE-1530 (3'-sialyllacto-N-neotetraose), K130
(5-[[4-[3-[[4-[(4-aminophenyl)sulfonyl]phenyl]amino]propoxy]-3,5-dimethox-
yphenyl]methyl]-2,4-pyrimidinediamine), PD 138312
((R)-7-[3-(1-amino-1-methylethyl)-1-pyrrolidinyl]-1-cyclopropyl-6-fluoro--
1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid), PD 140248
(7-[(3R)-3-[(1S)-1-aminoethyl]-1-pyrrolidinyl]-1-(2,4-difluorophenyl)-6-f-
luoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid), CP
111905
(5-deoxy-5-[[(2E)-3-[3-hydroxy-4-(2-propenyloxy)phenyl]-2-methyl-1-oxo-2--
propenyl]amino]-1,2-O-methylene-D-neo-inositol), sulopenem
((5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-[[(1R,3S)-tetrahydro-1-oxido-3-t-
hienyl]thio]-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic
acid), ritipenam acoxyl
((5R,6R)-3-[[(aminocarbonyl)oxy]methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-4-t-
hia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid
(acetyloxy)methyl ester), RO-65-5788
((6R,7R)-7-[[(2Z)-(5-amino-1,2,4-thiadiazol-3-yl)(hydroxyimino)acetyl]ami-
no]-3-[(E)-[(3'R)-1'-[[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy]carbonyl]-2-
-oxo[1,3'-bipyrrolidin]-3-ylidene]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]-
oct-2-ene-2-carboxylic acid monosodium salt), Sch-40832
(N-[[48-[1-[[2,6-dideoxy-3-O-(2,6-dideoxy-D-arabino-hexopyranosyl)-D-arab-
ino-hexopyranosyl]oxy]ethyl]-15-ethylidene-1,3a,4,5,10,11,12,13,14,15,19,2-
0,21,22,28,29,41,42-octadecahydro-41-hydroxy-12,45-bis(1-hydroxyethyl)-1-(-
hydroxymethyl)-22-(1-hydroxy-1-methylpropyl)-36-methyl-51,54,57-tris(methy-
lene)-3-(methylthio)-10,13,20,27,38,49,52,55,58-nonaoxo-18H,27H-5a,29-(imi-
noethaniminoethanimino ethaniminoethanimino[7,2]quinolinomethanoxy
methano)-9,6:19,16:26,23:33,30-tetranitrilo-16H,33aH-imidazo[1',5':1,6]py-
rido[3,2-m][1,11,17,24,4,7,20,27]tetrathiatetraazacyclotriacontin-1-yl]car-
bonyl]-2,3-didehydroalanyl-2,3-didehydro-alanine methyl ester
stereoisomer), micacocidin A
((OC-6-26-A)-[(4S)-2-[(2S)-2-[(2R,4R)-2-[(4R)-4,5-dihydro-2-[2-(hydroxy-.-
kappa.O)-6-pentylphenyl]-4-thiazolyl-.kappa.N3]-3-methyl-4-thiazolidinyl-.-
kappa.N3]-2-(hydroxy-.kappa.O)-1,1-dimethylethyl]-4,5-dihydro-4-methyl-4-t-
hiazolecarboxylato(2-)-.kappa.N3, .kappa.O4]-Zinc), SR-15402
((1S,5S,8aS,8bR)-1,2,5,6,7,8,8a,8b-octahydro-1-[(1R)-1-hydroxyethyl]-2-ox-
o-5-[(3S)-3-pyrrolidinylthio]-azeto[2,1-a]isoindole-4-carboxylic
acid), TOC 39
(1-(2-amino-2-oxoethyl)-4-[[(1E)-2-[(6R,7R)-7-[[(2Z)-(2-amino-4-th-
iazolyl)(hydroxyimino)acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.-
2.0]oct-2-en-3-yl]ethenyl]thio]-pyridinium inner salt), carumonam
([[(Z)-[2-[[(2S,3S)-2-[[(aminocarbonyl)oxy]methyl]-4-oxo-1-sulfo-3-azetid-
inyl]amino]-1-(2-amino-4-thiazolyl)-2-oxoethylidene]amino]oxy]-acetic
acid), cefozopran
(1-[[(6R,7R)-7-[[(2Z)-(5-amino-1,2,4-thiadiazol-3-yl)(methoxy
imino)acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3--
yl]methyl]-imidazo[1,2-b]pyridazinium inner salt), cefetamet
pivoxil ((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxy
imino)acetyl]amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2--
carboxylic acid (2,2-dimethyl-1-oxopropoxy)methyl ester), or T 3811
(des-F(6)-quinolone).
24. The method according to claim 22, wherein said second
antimicrobial agent is imipenen, amikacin, netilmicin, fosfomycin,
gentamicin, ceftriaxone, teicoplanin, Ziracin, LY333328, CL331022,
HMR3647, Linezolid, Synercid, Aztreonam or Metronidazole.
25. The method according to claim 17, wherein said subject is a
human or an animal.
26. The method according to claim 25, wherein said subject is a
human.
27. The compound of claim 1 having the formula (III): ##STR01751##
wherein R.sup.15 is hydrido or a carbamate amino protecting group;
and wherein R.sup.16 is ##STR01752##
28. The compound of claim 2 having the formula (III): ##STR01753##
wherein R.sup.15 is hydrido or a carbamate amino protecting group;
wherein R.sup.16 is ##STR01754## and wherein R.sup.57 is a halo or
halo substituted alkyl group.
29. The compound according to either of claim 27 or 28 wherein said
compound is TABLE-US-00008 Compound # R.sup.16 2 ##STR01755## 10
##STR01756## 25 ##STR01757##
Description
[0001] This application is a divisional of U.S. patent application
Ser. No. 09/737,908 filed Dec. 15, 2000, which claims priority to
U.S. Provisional Application No. 60/170,946, filed Dec. 15, 1999
and U.S. Provisional Application No. 60/208,222, filed May 30,
2000.
FIELD OF THE INVENTION
[0002] The present invention relates to novel lipopeptide
compounds. The invention also relates to pharmaceutical
compositions of these compounds and methods of using these
compounds as antibacterial compounds. The invention also relates to
methods of producing these novel lipopeptide compounds and
intermediates used in producing these compounds.
BACKGROUND OF THE INVENTION
[0003] The rapid increase in the incidence of gram-positive
infections--including those caused by resistant bacteria--has
sparked renewed interest in the development of novel classes of
antibiotics. A class of compounds which have shown potential as
useful antibiotics includes the A-21978C lipopeptides described in,
for example, U.S. Pat. Nos. RE 32,333; RE 32,455; RE 32,311; RE
32,310; 4,482,487; 4,537,717; and 5,912,226. Daptomycin, a member
of this class, has potent bactericidal activity in vitro and in
vivo against clinically relevant gram-positive bacteria that cause
serious and life-threatening diseases. These bacteria include
resistant pathogens, such as vancomycin-resistant enterococci
(VRE), methicillin-resistant Staphylococcus areus (MRSA),
glycopeptide intermediate susceptible Staphylococcus areus (GISA),
coagulase-negative staphylococci (CNS), and penicillin-resistant
Streptococcus pneumoniae (PRSP), for which there are few
therapeutic alternatives. See, e.g. Tally et al., 1999, Exp. Opin.
Invest. Drugs 8:1223-1238.
[0004] Despite the promise that antibacterial agents such as
daptomycin offer, the need for novel antibiotics continues. Many
pathogens have been repeatedly exposed to commonly-used
antibiotics. This exposure has led to the selection of variant
antibacterial strains resistant to a broad spectrum of antibiotics.
The loss of potency and effectiveness of an antibiotic caused by
resistant mechanisms renders the
antibiotic ineffective and consequently can lead to
life-threatening infections that are virtually untreatable. As new
antibiotics come to market pathogens may develop resistance or
intermediate resistance to these new drugs, effectively creating a
need for a stream of new antibacterial agents to combat these
emerging strains. In addition compounds that exhibit bacteriacidal
activity would offer advantages over present bacteriastatic
compounds. Thus, novel synthetic antibacterial agents would be
expected to be useful to treat not only "natural" pathogens, but
also intermediate drug resistant and drug resistant pathogens
because the pathogen has never been exposed to the novel
antibacterial agent. Additionally, new antibacterial agents may
exhibit differential effectiveness against different types of
pathogens.
SUMMARY OF THE INVENTION
[0005] The present invention addresses this problem by providing
novel lipopeptide compounds which have antibacterial activity
against a broad spectrum of bacteria, including drug-resistant
bacteria. Further, the compounds of the present invention exhibit
bacteriacidal activity.
[0006] The present invention comprises, in one aspect,
antibacterial compounds of Formula I:
##STR00001##
and salts thereof,
[0007] wherein R is:
##STR00002##
[0008] wherein X and X'' are independently selected from C.dbd.O,
C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X, S.dbd.O or SO.sub.2;
[0009] wherein n is 0 or 1;
[0010] wherein R.sup.X is selected from alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,
carboxy or carboalkoxy;
[0011] wherein B is X''R.sup.Y, H, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, cycloalkyl or heterocyclyl;
[0012] wherein R.sup.Y is selected from hydrido, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
hydroxyl;
[0013] wherein A is H, NH.sub.2, NHR.sup.A, NR.sup.AR.sup.B, alkyl,
alkenyl, alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or
heterocyclyl;
[0014] wherein R.sup.A and R.sup.B are independently selected from
alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl
or carboalkoxy;
[0015] wherein when n is 0, then A is additionally selected
from:
##STR00003##
[0016] wherein each of R.sup.50-R.sup.53 is independently selected
from C.sub.1-C.sub.15 alkyl;
[0017] alternatively, wherein B and A together form a 5-7 membered
heterocyclic or heteroaryl ring.
[0018] Wherein R.sup.1 is
##STR00004##
[0019] wherein X' and X''' are independently selected from C.dbd.O,
C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X', S.dbd.O or SO.sub.2;
[0020] wherein m is 0 or 1;
[0021] wherein R.sup.X' is selected from alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,
carboxy or carboalkoxy;
[0022] wherein B' is X'''R.sup.Y', H, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl or heterocyclyl; and
[0023] wherein R.sup.Y' is selected from hydrido, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
hydroxyl.
[0024] In one aspect of the invention, A' is H, NH.sub.2,
NHR.sup.A', NR.sup.A'R.sup.B', heteroaryl, cycloalkyl or
heterocyclyl;
[0025] wherein R.sup.A' and R.sup.B' are independently selected
from alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,
heterocyclyl or carboalkoxy;
[0026] wherein when m is 0, then A' is additionally selected
from:
##STR00005##
[0027] wherein each of R.sup.50-R.sup.53 is independently selected
from C.sub.1-C.sub.15 alkyl;
[0028] provided that when B' is H and X' is C.dbd.O, then A' is
other than
[0029] (a) a pyridinyl ring substituted with one substitutent
NHC(O)R.sup.D or
[0030] (b) a C.sub.5-C.sub.6 saturated cycloalkyl ring substituted
with one substitutent NHC(O)R.sup.D;
[0031] wherein R.sup.D is C.sub.1-C.sub.17 unsubstituted alkyl or
C.sub.2-C.sub.17 unsubstituted alkenyl; and when B' is H and m=0,
then A' is not H.
[0032] In another aspect of the invention, A' is aryl;
[0033] provided that when B' is H and X' is C.dbd.O, then A' is
other than a phenyl ring substituted with substitutent
NHC(O)R.sup.D, wherein R.sup.D is defined as above, which may be
further optionally substituted on the phenyl ring with 1-2
substituents independently selected from amino, nitro,
C.sub.1-C.sub.3 alkyl, hydroxyl, C.sub.1-C.sub.3 alkoxy, halo,
mercapto, C.sub.1-C.sub.3 alkylthio, carbamyl or C.sub.1-C.sub.3
alkyl carbamyl.
[0034] In a third aspect of the invention, A' is alkyl, alkenyl,
alkynyl, alkoxy or aryloxy;
[0035] provided that when B' is H and X' is C.dbd.O, then A' is
other than
[0036] (a) --(C.sub.1-C.sub.16unsubstituted alkyl)-NH.sub.2;
[0037] (b) --(C.sub.1-C.sub.10 unsubstituted alkyl)-NHC(O)R.sup.D,
wherein R.sup.D is defined as described above;
[0038] (c) --C.sub.1-C.sub.18 alkyl, optionally substituted with up
to one hydroxyl, carboxyl or C.sub.1-C.sub.3 alkoxy, or one to
three halo substituents;
[0039] (d) --C.sub.4-C.sub.18 unsubstituted alkenyl;
##STR00006##
[0040] wherein R.sup.54 is selected from
C.sub.1-C.sub.17-unsubstituted alkyl or
C.sub.2-C.sub.17-unsubstituted alkenyl; wherein R.sup.55 is
selected from hydroxyethyl, hydroxymethyl, mercaptomethyl,
mercaptoethyl, methylthioethyl, 2-thienyl, 3-indolemethyl, phenyl
optionally substituted with a group selected from halo, nitro,
C.sub.1-C.sub.3-unsubstituted alkyl, hydroxy,
C.sub.1-C.sub.3-unsubstituted alkoxy, C.sub.1-C.sub.3-unsubstituted
alkylthio, carbamyl or C.sub.1-C.sub.3 unsubstituted alkylcarbamyl;
or benzyl optionally substituted with a group selected from halo,
nitro, C.sub.1-C.sub.3-unsubstituted alkyl, hydroxy,
C.sub.1-C.sub.3-unsubstituted alkoxy, C.sub.1-C.sub.3-unsubstituted
alkylthio, carbamyl or C.sub.1-C.sub.3 unsubstituted alkylcarbamyl;
wherein t is 0 or 1 and wherein u is an integer from 1-3; and
[0041] when B' is H and X' is C.dbd.O, then X', together with A',
does not form a carbamate amino protecting group; and
[0042] when B' is H and m is 0, then A' is other than
C.sub.4-C.sub.14 unsubstituted alkyl.
[0043] In a fourth aspect of the invention, B' and A' together form
a 5-7 membered heterocyclic or heteroaryl ring.
[0044] Wherein R.sup.2 is
##STR00007##
wherein K and K' together form a C.sub.3-C.sub.7 cycloalkyl or
heterocyclyl ring or a C.sub.5-C.sub.10 aryl or heteroaryl
ring;
[0045] wherein J is selected from the group consisting of hydrido,
amino, NHR.sup.J, NR.sub.JR.sup.K, alkyl, alkenyl, alkynyl, alkoxy,
aryloxy, aryl, heteroaryl, cycloalkyl, heterocyclyl, alkylamino,
hydroxyl, thio, alkylthio, alkenylthio, sulfinyl, sulfonyl, azido,
cyano, halo,
##STR00008##
[0046] wherein each of R.sup.24, R.sup.25, and R.sup.26 is
independently selected from the group consisting of alkyl,
cycloalkyl, heterocyclyl, aryl and heteroaryl; or R.sup.24 and
R.sup.25 together form a 5-8 membered heterocyclyl ring;
[0047] wherein R.sup.J and R.sup.K are independently selected from
alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or
heterocyclyl; or
[0048] alternatively, wherein J, together with R.sup.17, forms a
5-8 membered heterocyclyl or cycloalkyl ring; or
[0049] alternatively, wherein J, together with both R.sup.17 and
R.sup.18, forms a 5-8 membered aryl, cycloalkyl, heterocyclyl or
heteroaryl ring; and
[0050] wherein each of R.sup.17 and R.sup.18 is independently
selected from the group consisting of hydrido, halo, hydroxyl,
alkoxy, amino, thio, sulfinyl, sulfonyl and
##STR00009##
[0051] wherein R.sup.17 and R.sup.18 taken together can form a
group consisting of ketal, thioketal,
##STR00010##
[0052] wherein each of R.sup.22 and R.sup.23 is independently
selected from the group consisting of hydrido and alkyl.
[0053] In another embodiment, the invention also provides
pharmaceutical compositions comprising compounds of Formula I and
methods of use thereof.
[0054] In a further embodiment, the invention provides methods of
making compounds of Formula I and pharmaceutical compositions
thereof.
[0055] In a further embodiment, the invention provides compounds
useful as intermediates for the preparation of compounds of Formula
I.
[0056] In a still further embodiment, the invention provides
methods of use of the compounds of Formula I to treat bacterial
infections in humans.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0057] Molecular terms, when used in this application, have their
common meaning unless otherwise specified.
[0058] The term "hydrido" denotes a single hydrogen atom (H).
[0059] The term "acyl" is defined as a carbonyl radical attached to
an alkyl, alkenyl, alkynyl, cycloalkyl, heterocycyl, aryl or
heteroaryl group, examples including, without limitation, such
radicals as acetyl and benzoyl.
[0060] The term "amino" denotes a nitrogen radical containing two
substituents independently selected from the group consisting of
hydrido, alkyl, cycloalkyl, carboalkoxy, heterocyclyl, aryl,
heteroaryl and sulfonyl. Subsets of the term amino are (1) the term
"unsubstituted amino" which denotes an NH.sub.2 radical, (2) the
term "mono substituted amino" which is defined as a nitrogen
radical containing a hydrido group and a substituent group selected
from alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, and (3)
the term "disubstituted amino" which is defined as a nitrogen
radical containing two substituent groups independently selected
from, alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.
Preferred mono substituted amino radicals are "lower mono
substituted amino" radicals, whereby the substituent group is a
lower alkyl group. Preferred disubstituted amino radicals are
"lower disubstituted amino" radicals, whereby the substituent
groups are lower alkyl.
[0061] The term "acyloxy" denotes an oxygen radical adjacent to an
acyl group.
[0062] The term "acylamino" denotes a nitrogen radical adjacent to
an acyl group.
[0063] The term "carboalkoxy" is defined as a carbonyl radical
adjacent to an alkoxy or aryloxy group.
[0064] The term "carboxyamido" denotes a carbonyl radical adjacent
to an amino group.
[0065] The term "halo" is defined as a bromo, chloro, fluoro or
iodo radical.
[0066] The term "thio" denotes a radical containing a substituent
group independently selected from hydrido, alkyl, cycloalkyl,
heterocyclyl, aryl and heteroaryl, attached to a divalent sulfur
atom, such as, methylthio and phenylthio.
[0067] The term "alkyl" is defined as a linear or branched,
saturated radical having one to about twenty carbon atoms unless
otherwise specified. Preferred alkyl radicals are "lower alkyl"
radicals having one to about five carbon atoms. One or more
hydrogen atoms can also be replaced by a substitutent group
selected from acyl, amino, acylamino, acyloxy, carboalkoxy,
carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio, alkyl,
alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,
alkoxy, aryloxy, sulfinyl, sulfonyl, oxo, guanidino, formyl and an
amino acid side chain. Examples of alkyl groups include, without
limitation, methyl, tert-butyl, isopropyl, and methoxymethyl.
Subsets of the term alkyl are (1) "unsubstituted alkyl" which is
defined as an alkyl group that bears no substituent groups (2)
"substituted alkyl" which denotes an alkyl radical in which (a) one
or more hydrogen atoms is replaced by a substitutent group selected
from acyl, acyloxy, carboalkoxy, carboxy, carboxyamido, cyano,
nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,
aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,
N-sulfonylcarboxyamido, N-acylaminosulfonyl or (b) two or more
hydrogen atoms are each replaced by a substituent group
independently selected from hydroxyl, carboxy, C.sub.1-C.sub.3
alkoxy, amino, acylamino, oxo or guanidino; and (3) the term
"selected substituted alkyl" which denotes an alkyl radical in
which (a) one proton is replaced by a group selected from hydroxyl,
carboxy C.sub.1-C.sub.3 alkoxy, unsubstituted amino, acylamino, or
acylamino phenyl or (b) one to three protons is replaced by a halo
substituent.
[0068] The term "alkenyl" is defined as linear or branched radicals
having two to about twenty carbon atoms, preferably three to about
ten carbon atoms, and containing at least one carbon-carbon double
bond. One or more hydrogen atoms can also be replaced by a
substituent group selected from acyl, amino, acylamino, acyloxy,
carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro,
thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, formyl, oxo and
guanidino. The double bond portion(s) of the unsaturated
hydrocarbon chain may be either in the cis or trans configuration.
Examples of alkenyl groups include, without limitation, ethylenyl
or phenyl ethylenyl.
[0069] The term "alkynyl" denotes linear or branched radicals
having from two to about ten carbon atoms, and containing at least
one carbon-carbon triple bond. One or more hydrogen atoms can also
be replaced by a substituent group selected from acyl, amino,
acylamino, acyloxy, carboalkoxy, carboxy, carboxyamido, cyano,
halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl,
heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl,
sulfonyl, formyl, oxo and guanidino. An example of alkynyl group
includes, without limitation, propynyl.
[0070] The term "aryl" or "aryl ring" denotes aromatic radicals in
a single or fused carbocyclic ring system, having from five to
fourteen ring members. In a preferred embodiment, the ring system
has from six to ten ring members. One or more hydrogen atoms may
also be replaced by a substituent group selected from acyl, amino,
acylamino, acyloxy, azido, alkylthio, carboalkoxy, carboxy,
carboxyamido, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl,
alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy,
aryloxy, sulfinyl, sulfonyl and formyl. Examples of aryl groups
include, without limitation, phenyl, naphthyl, biphenyl, terphenyl.
Subsets of the term aryl are (1) the term "phenyl" which denotes a
compound of the formula:
##STR00011##
(2) the term "substituted phenyl" which is defined as a phenyl
radical in which one or more protons are replaced by a substituent
group selected from acyl, amino, acyloxy, azido, alkylthio,
carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro,
thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,
N-sulfonylcarboxyamido, and N-acylaminosulfonyl and (3) the term
"acylamino phenyl" denotes a phenyl radical in which one hydrogen
atom is replaced by an acylamino group. One or more additional
hydrogen atoms can also be replaced by a substituent group selected
from acyl, amino, acylamino, acyloxy, azido, alkylthio,
carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro,
thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,
N-sulfonylcarboxyamido, and N-acylaminosulfonyl.
[0071] "Heteroaryl" or "heteroaryl ring" denotes an aromatic
radical which contain one to four hetero atoms or hetero groups
selected from O, N, S.
##STR00012##
in a single or fused heterocyclic ring system, having from five to
fifteen ring members. In a preferred embodiment, the heteroaryl
ring system has from six to ten ring members. One or more hydrogen
atoms may also be replaced by a substituent group selected from
acyl, amino, acylamino, acyloxy, carboalkoxy, carboxy,
carboxyamido, cyano, halo, hydroxyl, nitro, thio, thiocarbonyl,
alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.
Examples of heteroaryl groups include, without limitation,
pyridinyl, thiazolyl, thiadiazoyl, isoquinolinyl, pyrazolyl,
oxazolyl, oxadiazoyl, triazolyl, and pyrrolyl groups. Subsets of
the term heteroaryl are (1) the term "pyridinyl" which denotes
compounds of the formula:
##STR00013##
(2) the term "substituted pyridinyl" which is defined as a
pyridinyl radical in which one or more protons is replaced by a
substituent group selected from acyl, amino, acyloxy, carboalkoxy,
carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio, alkyl,
alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,
alkoxy, aryloxy, sulfinyl, sulfonyl, N-sulfonylcarboxyamido, and
N-acylaminosulfonyl and (3) the term "acylamino pyridinyl" which
denotes a pyridinyl radical in which one hydrogen atom is replaced
by an acylamino group, additionally, one or more additional
hydrogen atoms can also be replaced by a substituent group selected
from acyl, amino, acylamino, acyloxy, carboalkoxy, carboxy,
carboxyamido, cyano, halo, hydroxyl, nitro, thio, thiocarbonyl,
alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,
N-sulfonylcarboxyamido, and N-acylaminosulfonyl.
[0072] The term "cycloalkyl" or "cycloalkyl ring" is defined as a
saturated or partially unsaturated carbocyclic ring in a single or
fused carbocyclic ring system having from three to twelve ring
members. In a preferred embodiment, a cycloalkyl is a ring system
having three to seven ring members. One or more hydrogen atoms may
also be replaced by a substituent group selected from acyl, amino,
acylamino, acyloxy, carboalkoxy, carboxy, carboxyamido, cyano,
halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl,
heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl
and formyl. Examples of a cycloalkyl group include, without
limitation, cyclopropyl, cyclobutyl, cyclohexyl, and
cycloheptyl.
[0073] The term "heterocyclyl," "heterocyclic" or "heterocyclyl
ring" is defined as a saturated or partially unsaturated ring
containing one to four hetero atoms or hetero groups selected from
O, N, NH,
##STR00014##
wherein R.sup.Z is as defined for R.sup.X,
##STR00015##
in a single or fused heterocyclic ring system having from three to
twelve ring members. In a preferred embodiment, a heterocyclyl is a
ring system having three to seven ring members. One or more
hydrogen atoms may also be replaced by a substituent group selected
from acyl, amino, acylamino, acyloxy, oxo, thiocarbonyl, imino,
carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro,
thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,
heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl and formyl.
Examples of a heterocyclyl group include, without limitation,
morpholinyl, piperidinyl, and pyrrolidinyl.
[0074] The term "alkoxy" denotes oxy-containing radicals
substituted with an alkyl, cycloalkyl or heterocyclyl group.
Examples include, without limitation, methoxy, tert-butoxy,
benzyloxy and cyclohexyloxy.
[0075] The term "aryloxy" denotes oxy-containing radicals
substituted with an aryl or heteroaryl group. Examples include,
without limitation, phenoxy.
[0076] The term "amino acid side chain" denotes any side chain (R
group) from a naturally-occurring or a non-naturally occurring
amino acid.
[0077] The term "sulfinyl" is defined as a tetravalent sulfur
radical substituted with an oxo substituent and a second
substituent selected from the group consisting of alkyl,
cycloalkyl, heterocyclyl, aryl, or heteroaryl group.
[0078] The term "sulfonyl" is defined as a hexavalent sulfur
radical substituted with two oxo substituents and a third
substituent selected from alkyl, cycloalkyl, heterocyclyl aryl, or
heteroaryl.
[0079] The term "carbamate amino protecting group" is defined as a
recognized amino protecting group that when bound to an amino group
forms a carbamate. Examples of carbamate amino protecting groups
can be found in "Protective Groups in Organic Synthesis" by
Theodora W. Greene, John Wiley and Sons, New York, 1981. Examples
of carbamate amino protecting groups include benzyloxycarbonyl,
t-butoxycarbonyl, t-amyloxycarbonyl, isobornyloxycarbonyl,
adamantyloxycarbonyl, chlorobenzyloxycarbonyl,
nitrobenzyloxycarbonyl or the like.
[0080] The salts of the compounds of the invention (preferably a
compound of Formula I) include acid addition salts and base
addition salts. In a preferred embodiment, the salt is a
pharmaceutically acceptable salt of the compound of Formula I. The
term "pharmaceutically-acceptable salts" embraces salts commonly
used to form alkali metal salts and to form addition salts of free
acids or free bases. The nature of the salt is not critical,
provided that it is pharmaceutically-acceptable. Suitable
pharmaceutically-acceptable acid addition salts of the compounds of
the invention (preferably a compound of Formula I) may be prepared
from an inorganic acid or an organic acid. Examples of such
inorganic acids include, without limitation, hydrochloric,
hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric
acid. Appropriate organic acids may be selected from aliphatic,
cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic
and sulfonic classes of organic acids, examples of which include,
without limitation, formic, acetic, propionic, succinic, glycolic,
gluconic, maleic, embonic (pamoic), methanesulfonic,
ethanesulfonic, 2-hydroxyethanesulfonic, pantothenic,
benzenesulfonic, toluenesulfonic, sulfanilic, mesylic,
cyclohexylaminosulfonic, stearic, algenic, .beta.-hydroxybutyric,
malonic, galactic, and galacturonic acid. Suitable
pharmaceutically-acceptable base addition salts of compounds of the
invention (preferably a compound of Formula I) include, but are not
limited to, metallic salts made from aluminum, calcium, lithium,
magnesium, potassium, sodium and zinc or organic salts made from
N,N'-dibenzylethylenediamine, chloroprocaine, choline,
diethanolamine, ethylenediamine, N-methylglucamine, lysine and
procaine. All of these salts may be prepared by conventional means
from the corresponding compound of the invention (preferably a
compound of Formula I) by treating, for example, the compound of
the invention (preferably a compound of Formula I) with the
appropriate acid or base.
[0081] The compounds of the invention (preferably compounds of
Formula I) can possess one or more asymmetric carbon atoms and are
thus capable of existing in the form of optical isomers as well as
in the form of racemic or non-racemic mixtures thereof. The
compounds of the invention (preferably compounds of Formula I) can
be utilized in the present invention as a single isomer or as a
mixture of stereochemical isomeric forms. Diastereoisomers, i.e.,
nonsuperimposable stereochemical isomers, can be separated by
conventional means such as chromatography, distillation,
crystallization or sublimation. The optical isomers can be obtained
by resolution of the racemic mixtures according to conventional
processes, for example by formation of diastereoisomeric salts by
treatment with an optically active acid or base. Examples of
appropriate acids include, without limitation, tartaric,
diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and
camphorsulfonic acid. The mixture of diastereomers can be separated
by crystallization followed by liberation of the optically active
bases from these salts. An alternative process for separation of
optical isomers includes the use of a chiral chromatography column
optimally chosen to maximize the separation of the enantiomers.
Still another available method involves synthesis of covalent
diastereoisomeric molecules by reacting compounds of the invention
(preferably compounds of Formula I) with an optically pure acid in
an activated form or an optically pure isocyanate. The synthesized
diastereoisomers can be separated by conventional means such as
chromatography, distillation, crystallization or sublimation, and
then hydrolyzed to obtain the enantiomerically pure compound. The
optically active compounds of the invention (preferably compounds
of Formula I) can likewise be obtained by utilizing optically
active starting materials. These isomers may be in the form of a
free acid, a free base, an ester or a salt.
[0082] The invention also embraces isolated compounds. An isolated
compound refers to a compound which represents at least 10%,
preferably at least 20%, more preferably at least 50% and most
preferably at least 80% of the compound present in the mixture. In
a preferred embodiment, the compound, a pharmaceutically acceptable
salt thereof, or a pharmaceutical composition comprising the
compound exhibits a detectable (i.e. statistically significant)
antimicrobial activity when tested in conventional biological
assays such as those described herein.
Lipopeptide Compounds
[0083] A compound of the formula (I):
##STR00016##
and salts thereof,
[0084] wherein R is:
##STR00017##
[0085] wherein X and X'' are independently selected from C.dbd.O,
C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X, S.dbd.O or SO.sub.2;
[0086] wherein n is 0 or 1;
[0087] wherein R.sup.X is selected from alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,
carboxy or carboalkoxy;
[0088] wherein B is X''R.sup.Y, H, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, cycloalkyl or heterocyclyl;
[0089] wherein R.sup.Y is selected from hydrido, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
hydroxyl;
[0090] wherein A is H, NH.sub.2, NHR.sup.A, NR.sup.AR.sup.B, alkyl,
alkenyl, alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or
heterocyclyl;
[0091] wherein R.sup.A and R.sup.B are independently selected from
alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl
or carboalkoxy;
[0092] wherein when n is 0, then A is additionally selected
from:
##STR00018##
[0093] wherein each of R.sup.50-R.sup.53 is independently selected
from C.sub.1-C.sub.15 alkyl;
[0094] alternatively, wherein B and A together form a 5-7 membered
heterocyclic or heteroaryl ring.
[0095] Wherein R.sup.1 is
##STR00019##
[0096] wherein X' and X''' are independently selected from C.dbd.O,
C.dbd.S, C.dbd.NH, C.dbd.NR.sup.X', S.dbd.O or SO.sub.2;
[0097] wherein m is 0 or 1;
[0098] wherein Rx is selected from alkyl, alkenyl, alkynyl, aryl,
heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy, carboxy or
carboalkoxy;
[0099] wherein B' is X'''R.sup.Y', H, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, cycloalkyl or heterocyclyl; and
[0100] wherein R.sup.Y' is selected from hydrido, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or
hydroxyl.
[0101] In one aspect of the invention, A' is H, NH.sub.2,
NHR.sup.A', NR.sup.A'R.sup.B', heteroaryl, cycloalkyl or
heterocyclyl;
[0102] wherein R.sup.A' and R.sup.B' are independently selected
from alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,
heterocyclyl or carboalkoxy;
[0103] wherein when m is 0, then A' is additionally selected
from:
##STR00020##
[0104] wherein each of R.sup.50-R.sup.53 is independently selected
from C.sub.1-C.sub.15 alkyl;
[0105] provided that when B' is H and X' is C.dbd.O, then A' is
other than
[0106] (a) a pyridinyl ring substituted with one substitutent
NHC(O)R.sup.D or
[0107] (b) a C.sub.5-C.sub.6 saturated cycloalkyl ring substituted
with one substitutent NHC(O)R.sup.D;
[0108] wherein R.sup.D is C.sub.1-C.sub.17 unsubstituted alkyl or
C.sub.2-C.sub.17 unsubstituted alkenyl; and when B' is H and m=0,
then A' is not H.
[0109] In another aspect of the invention, A' is aryl;
[0110] provided that when B' is H and X' is C.dbd.O, then A' is
other than a phenyl ring substituted with substitutent
NHC(O)R.sup.D, wherein R.sup.D is defined as above, which may be
further optionally substituted on the phenyl ring with 1-2
substituents independently selected from amino, nitro,
C.sub.1-C.sub.3 alkyl, hydroxyl, C.sub.1-C.sub.3 alkoxy, halo,
mercapto, C.sub.1-C.sub.3 alkylthio, carbamyl or C.sub.1-C.sub.3
alkyl carbamyl.
[0111] In a third aspect of the invention, A' is alkyl, alkenyl,
alkynyl, alkoxy or aryloxy;
[0112] provided that when B' is H and X' is C.dbd.O, then A' is
other than
[0113] (a) --(C.sub.1-C.sub.16 unsubstituted alkyl)-NH.sub.2;
[0114] (b) --(C.sub.1-C.sub.10 unsubstituted alkyl)-NHC(O)R.sup.D,
wherein R.sup.D is defined as described above;
[0115] (c) --C.sub.1-C.sub.18 alkyl, optionally substituted with up
to one hydroxyl, carboxyl or C.sub.1-C.sub.3 alkoxy, or one to
three halo substituents;
[0116] (d) --C.sub.4-C.sub.18 unsubstituted alkenyl;
##STR00021##
[0117] wherein R.sup.54 is selected from
C.sub.1-C.sub.17-unsubstituted alkyl or
C.sub.2-C.sub.17-unsubstituted alkenyl; wherein R.sup.55 is
selected from hydroxyethyl, hydroxymethyl, mercaptomethyl,
mercaptoethyl, methylthioethyl, 2-thienyl, 3-indolemethyl, phenyl
optionally substituted with a group selected from halo, nitro,
C.sub.1-C.sub.3-unsubstituted alkyl, hydroxy,
C.sub.1-C.sub.3-unsubstituted alkoxy, C.sub.1-C.sub.3-unsubstituted
alkylthio, carbamyl or C.sub.1-C.sub.3 unsubstituted alkylcarbamyl;
or benzyl optionally substituted with a group selected from halo,
nitro, C.sub.1-C.sub.3-unsubstituted alkyl, hydroxy,
C.sub.1-C.sub.3-unsubstituted alkoxy, C.sub.1-C.sub.3-unsubstituted
alkylthio, carbamyl or C.sub.1-C.sub.3 unsubstituted alkylcarbamyl;
wherein t is 0 or 1 and wherein u is an integer from 1-3; and
[0118] when B is H and X is C.dbd.O, then X, together with A, does
not form a carbamate amino protecting group; and
[0119] when B' is H and m is 0, then A' is other than
C.sub.4-C.sub.14 unsubstituted alkyl.
[0120] In a fourth aspect of the invention, B' and A' together form
a 5-7 membered heterocyclic or heteroaryl ring.
[0121] Wherein R.sup.2 is
##STR00022##
[0122] wherein K and K' together form a C.sub.3-C.sub.7 cycloalkyl
or heterocyclyl ring or a C.sub.5-C.sub.10 aryl or heteroaryl
ring;
[0123] wherein J is selected from the group consisting of hydrido,
amino, NHR.sup.J, NR.sup.JR.sup.K, alkyl, alkenyl, alkynyl, alkoxy,
aryloxy, aryl, heteroaryl, cycloalkyl, heterocyclyl, alkylamino,
hydroxyl, thio, alkylthio, alkenylthio, sulfinyl, sulfonyl, azido,
cyano, halo,
##STR00023##
[0124] wherein each of R.sup.24, R.sup.25, and R.sup.26 is
independently selected from the group consisting of alkyl,
cycloalkyl, heterocyclyl, aryl and heteroaryl; or R.sup.24 and
R.sup.25 together form a 5-8 membered heterocyclyl ring;
[0125] wherein R.sup.J and R.sup.K are independently selected from
alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or
heterocyclyl; or
[0126] alternatively, wherein J, together with R.sup.17, forms a
5-8 membered heterocyclyl or cycloalkyl ring; or
[0127] alternatively, wherein J, together with both R.sup.17 and
R.sup.18, forms a 5-8 membered aryl, cycloalkyl, heterocyclyl or
heteroaryl ring; and
[0128] wherein each of R.sup.17 and R.sup.18 is independently
selected from the group consisting of hydrido, halo, hydroxyl,
alkoxy, amino, thio, sulfinyl, sulfonyl and
##STR00024##
[0129] wherein R.sup.17 and R.sup.18 taken together can form a
group consisting of ketal, thioketal,
##STR00025##
[0130] wherein each of R.sup.22 and R.sup.23 is independently
selected from the group consisting of hydrido and alkyl.
[0131] In a preferred embodiment of the invention, R is selected
from
##STR00026##
wherein each of R.sup.3, R.sup.4, R.sup.5, and R.sup.6 is
independently selected from the group consisting of hydrido, alkyl,
aryl, heterocyclyl and heteroaryl, and wherein R.sup.44 is selected
from the group consisting of alkyl, aryl, heterocyclyl and
heteroaryl.
[0132] In a more preferred embodiment of the invention R is
selected from
##STR00027##
wherein R.sup.4' is selected from the group consisting of alkyl,
aryl-substituted alkyl, substituted phenyl, heteroaryl,
heterocyclyl, optionally substituted (C.sub.8-C.sub.14)-straight
chain alkyl and
##STR00028##
[0133] wherein R.sup.7 is an alkyl group.
[0134] In an even more preferred embodiment of the invention, R
is
##STR00029##
wherein X.sup.3 is chloro or trifluoromethyl and wherein q is 0 or
1.
[0135] In a preferred embodiment of the invention, R.sup.1 is
selected from the group consisting of:
##STR00030##
wherein R.sup.8 is selected from an amino acid side chain, wherein
said amino acid side chain may be one that is naturally occurring
or one that is not naturally occurring, wherein each of R.sup.9,
R.sup.10 and R.sup.11 is selected from hydrido, alkyl, aryl,
heterocyclyl and heteroaryl; wherein R.sup.12 is selected from the
group consisting of heterocyclyl, heteroaryl, aryl, and alkyl and
wherein R.sup.13 is selected from (C.sub.1-C.sub.3)-alkyl and
aryl.
[0136] In a more preferred embodiment of the invention, R.sup.1 is
selected from the group consisting of
##STR00031##
wherein R.sup.8 is selected from tryptophan side chain and lysine
side chain; wherein each of R.sup.10 and R.sup.11 is independently
selected from hydrido and alkyl; wherein R.sup.12 is selected from
imidazolyl, N-methylimidazolyl, indolyl, quinolinyl,
benzyloxybenzyl, and benzylpiperidenylbenzyl; and wherein X.sup.4
is selected from fluoro and trifluoromethyl.
[0137] In a preferred embodiment of R.sup.2, J is selected from the
group consisting of hydrido, amino, azido and
##STR00032##
wherein R.sup.17 and R.sup.18 taken together form a group selected
from the group consisting of ketal,
##STR00033##
alternatively, R.sup.17 is hydroxyl when R.sup.18 is hydrido.
Alternatively, wherein J, together with R.sup.17, forms a
heterocyclyl ring.
[0138] In a more preferred embodiment of the invention, R.sup.2 is
selected from
##STR00034##
[0139] wherein R.sup.17 and R.sup.18 taken together form a group
selected from
##STR00035##
wherein R.sup.22 is selected from the group consisting of H and
alkyl; wherein R.sup.19 is selected from the group consisting of
hydrido, amino, azido and
##STR00036##
[0140] In an even more preferred embodiment of the invention
R.sup.2 is
##STR00037##
[0141] Another aspect of the present invention provides compounds
of formula (I), wherein R is selected from
NHCO--[(C.sub.6-C.sub.14)-alkyl]CH.sub.3, and R.sup.1 and R.sup.2
are selected from Table A below. More preferably, R is selected
from NHCO--[(CH.sub.2).sub.6-14]--CH.sub.3.
TABLE-US-00001 TABLE A R.sup.1 R.sup.2 ##STR00038## ##STR00039##
##STR00040## ##STR00041## NHSO.sub.2Ph ##STR00042## ##STR00043##
##STR00044## ##STR00045## ##STR00046## ##STR00047## ##STR00048##
##STR00049## ##STR00050## ##STR00051## ##STR00052## ##STR00053##
##STR00054## ##STR00055## ##STR00056## ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063##
##STR00064## ##STR00065## ##STR00066## ##STR00067## ##STR00068##
##STR00069## ##STR00070## ##STR00071## ##STR00072## ##STR00073##
##STR00074## ##STR00075## ##STR00076## ##STR00077## ##STR00078##
##STR00079## ##STR00080## ##STR00081## ##STR00082## ##STR00083##
##STR00084## ##STR00085## ##STR00086## ##STR00087## ##STR00088##
##STR00089## ##STR00090## ##STR00091## ##STR00092## ##STR00093##
##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098##
##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103##
##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108##
##STR00109## ##STR00110## ##STR00111## ##STR00112## ##STR00113##
##STR00114## ##STR00115## ##STR00116## ##STR00117## ##STR00118##
##STR00119## ##STR00120## ##STR00121## ##STR00122## ##STR00123##
##STR00124## ##STR00125## ##STR00126## ##STR00127## ##STR00128##
##STR00129## ##STR00130## ##STR00131## ##STR00132## ##STR00133##
##STR00134## ##STR00135## ##STR00136## ##STR00137## ##STR00138##
##STR00139## ##STR00140## ##STR00141## ##STR00142## ##STR00143##
##STR00144## ##STR00145## ##STR00146## ##STR00147## ##STR00148##
##STR00149## ##STR00150## ##STR00151## ##STR00152## ##STR00153##
##STR00154## ##STR00155## ##STR00156## ##STR00157## ##STR00158##
##STR00159## ##STR00160## ##STR00161## ##STR00162## ##STR00163##
##STR00164## ##STR00165## ##STR00166## ##STR00167## ##STR00168##
##STR00169## ##STR00170## ##STR00171## ##STR00172## ##STR00173##
##STR00174## ##STR00175## ##STR00176## ##STR00177## ##STR00178##
##STR00179## ##STR00180## NH(CH.sub.2).sub.2OH ##STR00181##
##STR00182## ##STR00183## ##STR00184## ##STR00185## ##STR00186##
##STR00187## ##STR00188## ##STR00189## ##STR00190## ##STR00191##
##STR00192## ##STR00193## ##STR00194## ##STR00195## ##STR00196##
##STR00197## ##STR00198## ##STR00199## ##STR00200## ##STR00201##
##STR00202## ##STR00203## ##STR00204## ##STR00205## ##STR00206##
##STR00207## ##STR00208## ##STR00209## ##STR00210## ##STR00211##
##STR00212## ##STR00213## ##STR00214## ##STR00215## ##STR00216##
##STR00217## ##STR00218## ##STR00219## ##STR00220## ##STR00221##
##STR00222## ##STR00223## ##STR00224## ##STR00225## ##STR00226##
##STR00227## ##STR00228## ##STR00229## ##STR00230## ##STR00231##
##STR00232## ##STR00233## ##STR00234## ##STR00235## ##STR00236##
##STR00237## ##STR00238## ##STR00239## ##STR00240## ##STR00241##
##STR00242## ##STR00243## ##STR00244## ##STR00245## ##STR00246##
##STR00247## ##STR00248## ##STR00249## ##STR00250## ##STR00251##
##STR00252## ##STR00253## ##STR00254## ##STR00255## ##STR00256##
##STR00257## ##STR00258## ##STR00259## ##STR00260## ##STR00261##
##STR00262## ##STR00263## ##STR00264## ##STR00265## ##STR00266##
##STR00267## ##STR00268## ##STR00269## ##STR00270## ##STR00271##
##STR00272## ##STR00273## ##STR00274## ##STR00275## ##STR00276##
##STR00277## ##STR00278## ##STR00279## ##STR00280##
##STR00281##
##STR00282## ##STR00283## ##STR00284## ##STR00285## ##STR00286##
##STR00287## ##STR00288## ##STR00289## ##STR00290## ##STR00291##
##STR00292## ##STR00293## ##STR00294## ##STR00295## ##STR00296##
##STR00297## ##STR00298## ##STR00299## ##STR00300## ##STR00301##
##STR00302## ##STR00303## ##STR00304## ##STR00305## ##STR00306##
##STR00307## ##STR00308## ##STR00309## ##STR00310## ##STR00311##
##STR00312## ##STR00313## ##STR00314## ##STR00315## ##STR00316##
##STR00317## ##STR00318## ##STR00319## ##STR00320## ##STR00321##
##STR00322## ##STR00323## ##STR00324## ##STR00325## ##STR00326##
##STR00327## ##STR00328## ##STR00329## ##STR00330## ##STR00331##
##STR00332## ##STR00333## ##STR00334## ##STR00335## ##STR00336##
##STR00337## ##STR00338## ##STR00339## ##STR00340## ##STR00341##
##STR00342## ##STR00343## ##STR00344## ##STR00345## ##STR00346##
##STR00347## ##STR00348## ##STR00349## ##STR00350## ##STR00351##
##STR00352## ##STR00353## ##STR00354## ##STR00355## ##STR00356##
##STR00357## ##STR00358## ##STR00359## ##STR00360## ##STR00361##
##STR00362## ##STR00363## ##STR00364## ##STR00365## ##STR00366##
##STR00367## ##STR00368## ##STR00369## ##STR00370## ##STR00371##
##STR00372## ##STR00373## ##STR00374## ##STR00375## ##STR00376##
##STR00377## ##STR00378## ##STR00379## ##STR00380## ##STR00381##
##STR00382## ##STR00383## ##STR00384## ##STR00385## ##STR00386##
##STR00387## ##STR00388## ##STR00389## ##STR00390## ##STR00391##
##STR00392## ##STR00393## ##STR00394## ##STR00395## ##STR00396##
##STR00397## ##STR00398## ##STR00399## ##STR00400## ##STR00401##
##STR00402## ##STR00403## ##STR00404## ##STR00405## ##STR00406##
##STR00407## ##STR00408## ##STR00409## ##STR00410## ##STR00411##
##STR00412## ##STR00413## ##STR00414## ##STR00415## ##STR00416##
##STR00417## ##STR00418## ##STR00419## ##STR00420## ##STR00421##
##STR00422## ##STR00423## ##STR00424## ##STR00425## ##STR00426##
##STR00427## ##STR00428## ##STR00429## ##STR00430## ##STR00431##
##STR00432## ##STR00433## ##STR00434## ##STR00435## ##STR00436##
##STR00437## ##STR00438## ##STR00439## ##STR00440## ##STR00441##
##STR00442## ##STR00443## ##STR00444## ##STR00445## ##STR00446##
##STR00447## ##STR00448## ##STR00449## ##STR00450## ##STR00451##
##STR00452## ##STR00453## ##STR00454## ##STR00455## ##STR00456##
##STR00457## ##STR00458## ##STR00459## ##STR00460## ##STR00461##
##STR00462## ##STR00463## ##STR00464## ##STR00465## ##STR00466##
##STR00467## ##STR00468## ##STR00469## ##STR00470## ##STR00471##
##STR00472## ##STR00473## ##STR00474## ##STR00475## ##STR00476##
##STR00477## ##STR00478## ##STR00479## ##STR00480## ##STR00481##
##STR00482## ##STR00483## ##STR00484## ##STR00485## ##STR00486##
##STR00487## ##STR00488## ##STR00489## ##STR00490## ##STR00491##
##STR00492## ##STR00493## ##STR00494## ##STR00495## ##STR00496##
##STR00497## ##STR00498## ##STR00499## ##STR00500## ##STR00501##
##STR00502## ##STR00503## ##STR00504## ##STR00505## ##STR00506##
##STR00507## ##STR00508## ##STR00509## ##STR00510## ##STR00511##
##STR00512## ##STR00513## ##STR00514## ##STR00515## ##STR00516##
##STR00517## ##STR00518## ##STR00519## ##STR00520## ##STR00521##
##STR00522## ##STR00523## ##STR00524## ##STR00525## ##STR00526##
##STR00527## ##STR00528## ##STR00529## ##STR00530## ##STR00531##
##STR00532## ##STR00533##
##STR00534## ##STR00535## ##STR00536## ##STR00537## ##STR00538##
##STR00539## ##STR00540## ##STR00541## ##STR00542## ##STR00543##
##STR00544## ##STR00545## ##STR00546## ##STR00547## ##STR00548##
##STR00549## ##STR00550## ##STR00551## ##STR00552## ##STR00553##
##STR00554## ##STR00555## ##STR00556## ##STR00557## ##STR00558##
##STR00559## ##STR00560## ##STR00561## ##STR00562## ##STR00563##
##STR00564## ##STR00565## ##STR00566## ##STR00567## ##STR00568##
##STR00569## ##STR00570## ##STR00571## ##STR00572## ##STR00573##
##STR00574## ##STR00575## ##STR00576## ##STR00577## ##STR00578##
##STR00579## ##STR00580## ##STR00581## ##STR00582## ##STR00583##
##STR00584## ##STR00585## ##STR00586## ##STR00587## ##STR00588##
##STR00589## ##STR00590## ##STR00591## ##STR00592## ##STR00593##
##STR00594## ##STR00595## ##STR00596## ##STR00597## ##STR00598##
##STR00599## ##STR00600## ##STR00601## ##STR00602## ##STR00603##
##STR00604## ##STR00605## ##STR00606## ##STR00607## ##STR00608##
##STR00609## ##STR00610## ##STR00611## ##STR00612## ##STR00613##
##STR00614## ##STR00615## ##STR00616## ##STR00617## ##STR00618##
##STR00619## ##STR00620## ##STR00621## ##STR00622## ##STR00623##
##STR00624## ##STR00625## ##STR00626## ##STR00627## ##STR00628##
##STR00629## ##STR00630## ##STR00631## ##STR00632## ##STR00633##
##STR00634## ##STR00635## ##STR00636## ##STR00637## ##STR00638##
##STR00639## ##STR00640## ##STR00641## ##STR00642## ##STR00643##
##STR00644## ##STR00645## ##STR00646## ##STR00647## ##STR00648##
##STR00649## ##STR00650## ##STR00651## ##STR00652## ##STR00653##
##STR00654## ##STR00655## ##STR00656## ##STR00657## ##STR00658##
##STR00659## ##STR00660## ##STR00661## ##STR00662## ##STR00663##
##STR00664## ##STR00665## ##STR00666## ##STR00667##
TABLE-US-00002 TABLE I Table I provides exemplary compounds of
Formula I: Synth Mass Ex Cpd # R R.sup.1 R.sup.2 Spec # 1
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00668## ##STR00669## 1863 6 2
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00670## ##STR00671## 1663 6 3
NHCO(CH.sub.2).sub.8CH.sub.3 NHSO.sub.2Ph ##STR00672## 1762 5 4
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00673## ##STR00674## 1792 4 5
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00675## ##STR00676## 1694 4 6
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00677## ##STR00678## 1722 4 7
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00679## ##STR00680## 1764 4 8
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00681## ##STR00682## 1720 4 9
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00683## ##STR00684## 1775 4 10
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00685## ##STR00686## 1740 2 11
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00687## ##STR00688## 1775 2 12
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00689## ##STR00690## 1820 2 13
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00691## ##STR00692## 1755 2 14
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00693## ##STR00694## 1755 2 15
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00695## ##STR00696## 1771 2 16
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00697## ##STR00698## 1771 2 17
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00699## ##STR00700## 1775 2 18
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00701## ##STR00702## 1812 3b 19
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00703## ##STR00704## 1785 2 20
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00705## ##STR00706## 1755 2 21
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00707## ##STR00708## 1756 3b 22
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00709## ##STR00710## 1757 2 23
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00711## ##STR00712## 1742 2 24
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00713## ##STR00714## 1790 2 25
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00715## ##STR00716## 1758 2 26
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00717## ##STR00718## 1758 2 27
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00719## ##STR00720## 1758 2 28
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00721## ##STR00722## 1726 3b 29
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00723## ##STR00724## 1728 3b 30
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00725## ##STR00726## 1741 3b 31
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00727## ##STR00728## 1741 3b 32
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00729## ##STR00730## 1771 3b 33
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00731## ##STR00732## 1851 3b 34
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00733## ##STR00734## 1767 3b 35
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00735## ##STR00736## 1782 3b 36
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00737## ##STR00738## 1780 8 37
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00739## ##STR00740## 1873 8 38
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00741## ##STR00742## 1729 1 39
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00743## ##STR00744## 1838 3b 40
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00745## ##STR00746## 1741 1 41
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00747## ##STR00748## 1908 3 42
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00749## ##STR00750## 1865 3 43
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00751## ##STR00752## 1893 3 44
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00753## ##STR00754## 1908 3 45
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00755## ##STR00756## 1808 3 46
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00757## ##STR00758## 1764 3 47
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00759## ##STR00760## 1750 3 48
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00761## ##STR00762## 1736 3 49
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00763## ##STR00764## 2004 3a 50
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00765## ##STR00766## 1712 1 51
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00767## ##STR00768## 1904 3a 52
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00769## ##STR00770## 1725 1 54
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00771## ##STR00772## 1749 3a 55
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00773## ##STR00774## 1884 3 56
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00775## ##STR00776## 1785 3 57
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00777## ##STR00778## 1853 3 58
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00779## ##STR00780## 1847 3 60
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00781## ##STR00782## 1778 3 61
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00783## ##STR00784## 1792 3 62
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00785## ##STR00786## 1826 3 63
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00787## ##STR00788## 1826 3 64
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00789## ##STR00790## 1838 3 65
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00791## ##STR00792## 1812 3 66
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00793## ##STR00794## 1808 3 67
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00795## ##STR00796## 1769 3 68
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00797## ##STR00798## 1824 3 69
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00799## ##STR00800## 1775 3 70
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00801## ##STR00802## 1820 3 72
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00803## ##STR00804## 1707 3 73
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00805## ##STR00806## 1758 3 74
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00807## ##STR00808## 1959 3 75
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00809## ##STR00810## 1810 3 76
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00811## ##STR00812## 1787 1g 77
NHCO(CH.sub.2).sub.8CH.sub.3 NH(CH.sub.2).sub.2OH ##STR00813## 1665
1 78 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00814## ##STR00815## 1820 1
79 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00816## ##STR00817## 1750 1 80
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00818## ##STR00819## 1779 1 81
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00820## ##STR00821## 1767 1e 82
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00822## ##STR00823## 1763 1 83
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00824## ##STR00825## 1869 1 84
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00826## ##STR00827## 1764 1 85
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00828## ##STR00829## 1714 1c 86
##STR00830## ##STR00831## ##STR00832## 1935 9 87
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00833## ##STR00834## 1863 1 88
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00835## ##STR00836## 2151 1 89
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00837## ##STR00838## 1887 1 90
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00839## ##STR00840## 2046 1 91
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00841## ##STR00842## 1996 1 92
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00843## ##STR00844## 1809 1 93
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00845## ##STR00846## 1783 1 94
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00847## ##STR00848## 1770 1 95
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00849## ##STR00850## 1836 1 96
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00851## ##STR00852## 1792 1 97
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00853## ##STR00854## 1847 1 98
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00855## ##STR00856## 1838 1 99
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00857## ##STR00858## 1837 1 100
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00859## ##STR00860## 1817 1 101
##STR00861## ##STR00862## ##STR00863## 1867 9 102
NHCO(CH.sub.2).sub.11CH.sub.3 ##STR00864## ##STR00865## 1849 9 103
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00866## ##STR00867## 1885 1 104
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00868## ##STR00869## 2150 1 105
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00870## ##STR00871## 1756 1 106
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00872## ##STR00873## 1833 1 107
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00874## ##STR00875## 1871 1 108
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00876## ##STR00877## 1873 1 109
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00878## ##STR00879## 1872 1 110
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00880## ##STR00881## 2014 1 111
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00882## ##STR00883## 1817 1 112
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00884## ##STR00885## 2121 1 113
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00886## ##STR00887## 2036 1 114
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00888## ##STR00889## 1826 1 115
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00890## ##STR00891## 1736 1 116
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00892## ##STR00893## 1797 1 117
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00894## ##STR00895## 1860 1 118
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00896## ##STR00897## 2055 1 119
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00898## ##STR00899## 1837 1 120
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00900## ##STR00901## 2104 1 121
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00902## ##STR00903## 1803 1 122
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00904## ##STR00905## 1755 1 123
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00906## ##STR00907## 1812 1 124
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00908## ##STR00909## 2002 1
125 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00910## ##STR00911## 1946 1
126 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00912## ##STR00913## 1918 1
127 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00914## ##STR00915## 1811 1
128 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00916## ##STR00917## 2050 1
129 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00918## ##STR00919## 1756 1
130 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00920## ##STR00921## 1762 1
131 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00922## ##STR00923## 1904 1
132 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00924## ##STR00925## 1962 1
133 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00926## ##STR00927## 1726 1
134 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00928## ##STR00929## 2074 1
135 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00930## ##STR00931## 1729 1
136 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00932## ##STR00933## 1729 1
137 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00934## ##STR00935## 2014 1
138 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00936## ##STR00937## 1762 1
139 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00938## ##STR00939## 1751 1
140 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00940## ##STR00941## 1881 1
141 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00942## ##STR00943## 1914 1
142 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00944## ##STR00945## 1753 1
143 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00946## ##STR00947## 1803 1
144 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00948## ##STR00949## 1813 1
145 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00950## ##STR00951## 2006 1
146 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00952## ##STR00953## 1701 1
147 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00954## ##STR00955## 1799 1
148 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00956## ##STR00957## 1978 1
149 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00958## ##STR00959## 1834 1
150 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00960## ##STR00961## 1777 1
151 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00962## ##STR00963## 1847 1
152 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00964## ##STR00965## 2074 1
153 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00966## ##STR00967## 1895 1
154 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00968## ##STR00969## 1867 1
155 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00970## ##STR00971## 1839 1
156 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00972## ##STR00973## 1781 1
157 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00974## ##STR00975## 1780 1
158 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00976## ##STR00977## 1781 1
159 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00978## ##STR00979## 1805 1
160 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00980## ##STR00981## 1990 1
161 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00982## ##STR00983## 1785 1
162 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00984## ##STR00985## 2092 1
163 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00986## ##STR00987## 1944 1
164 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00988## ##STR00989## 1817 1
165 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00990## ##STR00991## 2014 1
166 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00992## ##STR00993## 1747 1
167 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00994## ##STR00995## 1853 1
168 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00996## ##STR00997## 1762 1
169 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR00998## ##STR00999## 1829 1
171 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01000## ##STR01001## 1914 1
172 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01002## ##STR01003## 1767 1
173 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01004## ##STR01005## 1736 1
174 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01006## ##STR01007## 1718 1
175 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01008## ##STR01009## 1808 1
176 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01010## ##STR01011## 1781 1
177 NH.sub.2 ##STR01012## ##STR01013## 1632 1 178
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01014## ##STR01015## 1783 3 179
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01016## ##STR01017## 1884 3 180
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01018## ##STR01019## 1905 3 181
NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01020## ##STR01021## 1851 9
182 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01022## ##STR01023## 1801 3b
183 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01024## ##STR01025## 1833 1
184 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01026## ##STR01027## 1727 1
185 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01028## ##STR01029## 1743 1
186 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01030## ##STR01031## 1890 1
187 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01032## ##STR01033## 1756 1
189 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01034## ##STR01035## 1717 3b
190 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01036## ##STR01037## 1805 2
192 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01038## ##STR01039## 1811 8
193 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01040## ##STR01041## 1836 3
194 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01042## ##STR01043## 1795 1
195 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01044## ##STR01045## 1862 1
196 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01046## ##STR01047## 1780 1
197 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01048## ##STR01049## 1746 1
198 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01050## ##STR01051## 1754 1
199 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01052## ##STR01053## 1780 1
200 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01054## ##STR01055## 1792 8a
201 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01056## ##STR01057## 1821 1
202 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01058## ##STR01059## 1 203
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01060## ##STR01061## 1793 1 204
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01062## ##STR01063## 1893 205
NH(CH.sub.2).sub.8CH.sub.3 ##STR01064## ##STR01065## 1779 9a 206
NHCO(CH.sub.2).sub.8CO.sub.2Me ##STR01066## ##STR01067## 1851 9 207
NHCO(CH.sub.2).sub.6CO.sub.2Me ##STR01068## ##STR01069## 1823 9 208
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01070## ##STR01071## 1878 1 209
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01072## ##STR01073## 1880 1h 210
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01074## ##STR01075## 1851 1 211
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01076## ##STR01077## 1924 1 212
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01078## ##STR01079## 1701 1d 213
NHCO(CH.sub.2).sub.6NHBoc ##STR01080## ##STR01081## 1980 9 214
NHCO(CH.sub.2).sub.7NHBoc ##STR01082## ##STR01083## 1994 9 215
NHCO(CH.sub.2).sub.10NHBoc ##STR01084## ##STR01085## 2036 9 216
NHCO(CH.sub.2).sub.11NHBoc ##STR01086## ##STR01087## 2050 9 217
NHCO(CH.sub.2).sub.10NH.sub.2 ##STR01088## ##STR01089## 1836 9 218
NHCO(CH.sub.2).sub.11NH.sub.2 ##STR01090## ##STR01091## 1850 9 219
NHCO(CH.sub.2).sub.6CH(CH.sub.3).sub.2 ##STR01092## ##STR01093##
1807 9 220 NHCONH(CH.sub.2).sub.11CH.sub.3 ##STR01094##
##STR01095## 1865 9 221 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01096##
##STR01097## 1807 6 222 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01098##
##STR01099## 1935 1 223 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01100##
##STR01101## 1779 1 224 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01102##
##STR01103## 1936 1 225 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01104##
##STR01105## 1735 1 226 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01106##
##STR01107## 1958 1 227 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01108##
##STR01109## 1899 1 228 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01110##
##STR01111## 1917 1 229 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01112##
##STR01113## 1914 1 230 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01114##
##STR01115## 1969 1 231 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01116##
##STR01117## 1990 1 232 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01118##
##STR01119## 1940 1 233 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01120##
##STR01121## 1902 1 234 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01122##
##STR01123## 1901 1 235 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01124##
##STR01125## 1934 1 236 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01126##
##STR01127## 1984 1 237 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01128##
##STR01129## 1926 1 238 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01130##
##STR01131## 1944 1 239 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01132##
##STR01133## 1940 1 240 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01134##
##STR01135## 1995 1 241 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01136##
##STR01137## 2016 1 242 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01138##
##STR01139## 1928 1 243 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01140##
##STR01141## 1927 1 244 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01142##
##STR01143## 1960 1 245 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01144##
##STR01145## 1790 3 246 ##STR01146## ##STR01147## ##STR01148## 1807
9 247 ##STR01149## ##STR01150## ##STR01151## 1841 9 248
##STR01152## ##STR01153## ##STR01154## 1864 9 249 ##STR01155##
##STR01156## ##STR01157## 1843 9 250 ##STR01158## ##STR01159##
##STR01160## 1882 9 251 ##STR01161## ##STR01162## ##STR01163## 1823
9 252 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01164## ##STR01165## 1931
1
253 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01166## ##STR01167## 1886 1f
254 NHCO(CH.sub.2).sub.7CH.sub.3 ##STR01168## ##STR01169## 1650 7
255 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01170## ##STR01171## 1678 7
256 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01172## ##STR01173## 1692 7
257 NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01174## ##STR01175## 1706 7
258 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01176## ##STR01177## 1720 7a
259 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01178## ##STR01179## 1706 6
260 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01180## ##STR01181## 1678 7
261 NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01182## ##STR01183## 1705 7
262 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01184## ##STR01185## 1719 7a
263 ##STR01186## ##STR01187## ##STR01188## 1738 7 264 ##STR01189##
##STR01190## ##STR01191## 1862 9 265 NHCO(CH.sub.2).sub.8CH.sub.3
##STR01192## ##STR01193## 1890 1 266 NHCO(CH.sub.2).sub.8CH.sub.3
##STR01194## ##STR01195## 1841 1 267 NHCO(CH.sub.2).sub.8CH.sub.3
##STR01196## ##STR01197## 1910 1 268 NHCO(CH.sub.2).sub.8CH.sub.3
##STR01198## ##STR01199## 1940 9 269 ##STR01200## ##STR01201##
##STR01202## 1862 6 270 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01203##
##STR01204## 1706 6 271 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01205##
##STR01206## 1851 1 272 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01207##
##STR01208## 2081 1 273 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01209##
##STR01210## 1964 1 274 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01211##
##STR01212## 1793 1 275 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01213##
##STR01214## 1797 1 276 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01215##
##STR01216## 1973 1 277 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01217##
##STR01218## 1778 1 278 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01219##
##STR01220## 1780 1 279 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01221##
##STR01222## 1940 1 280 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01223##
##STR01224## 1797 1 281 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01225##
##STR01226## 1974 1 282 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01227##
##STR01228## 1807 1a 283 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01229##
##STR01230## 1797 1 284 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01231##
##STR01232## 1973 1 285 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01233##
##STR01234## 1796 1b 286 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01235##
##STR01236## 1898 1 287 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01237##
##STR01238## 1806 1 288 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01239##
##STR01240## 1812 1 289 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01241##
##STR01242## 1806 1 290 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01243##
##STR01244## 1806 1 291 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01245##
##STR01246## 1848 1 292 ##STR01247## ##STR01248## ##STR01249## 1738
7 293 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01250## ##STR01251## 1692
7 294 NHCO(CH.sub.2).sub.7CH.sub.3 ##STR01252## ##STR01253## 1650 7
295 NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01254## ##STR01255## 1991
10b 296 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01256## ##STR01257##
1978 10b 297 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01258## ##STR01259##
1964 10b 298 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01260##
##STR01261## 1950 10b 299 NHCONH(CH.sub.2).sub.10CH.sub.3
##STR01262## ##STR01263## 1992 10b 300
NHCONH(CH.sub.2).sub.11CH.sub.3 ##STR01264## ##STR01265## 2006 10b
301 NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01266## ##STR01267## 1791
10b 302 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01268## ##STR01269##
1778 10b 303 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01270## ##STR01271##
1764 10b 304 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01272##
##STR01273## 1750 10b 305 NHCONH(CH.sub.2).sub.10CH.sub.3
##STR01274## ##STR01275## 1792 10b 306
NHCONH(CH.sub.2).sub.11CH.sub.3 ##STR01276## ##STR01277## 1806 10b
307 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01278## ##STR01279## 1922 10b
308 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01280## ##STR01281## 1936
10b 309 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01282## ##STR01283##
1836 10b 310 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01284## ##STR01285##
1821 10b 311 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01286##
##STR01287## 1808 10b 312 NHCONH(CH.sub.2).sub.7CH.sub.3
##STR01288## ##STR01289## 1759 10b 313
NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01290## ##STR01291## 1665 7 314
NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01292## ##STR01293## 1707 7
315 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01294## ##STR01295## 1779
10a 316 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01296## ##STR01297##
1700 10a 317 NHCONH(CH.sub.2).sub.7CH.sub.3 ##STR01298##
##STR01299## 1806 10a 318 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01300##
##STR01301## 1793 10a 319 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01302##
##STR01303## 1714 10a 320 NHCO(CH.sub.2).sub.11CH.sub.3
##STR01304## ##STR01305## 1821 10a 321
NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01306## ##STR01307## 1848 10a
322 NHCO(CH.sub.2).sub.11CH.sub.3 ##STR01308## ##STR01309## 1742
10a 323 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01310## ##STR01311## 1943
1 324 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01312## ##STR01313## 2010 1
325 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01314## ##STR01315## 1893 1
326 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01316## ##STR01317## 956 1
327 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01318## ##STR01319## 1875 1
328 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01320## ##STR01321## 1919 1
329 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01322## ##STR01323## 1987 1
330 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01324## ##STR01325## 1909 1
331 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01326## ##STR01327## 1998 1
332 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01328## ##STR01329## 1807
10a 333 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01330## ##STR01331##
1834 10a 334 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01332##
##STR01333## 1728 10a 335 NHCONH(CH.sub.2).sub.11CH.sub.3
##STR01334## ##STR01335## 1757 10a 336
NHCONH(CH.sub.2).sub.11CH.sub.3 ##STR01336## ##STR01337## 1864 10a
337 NHCONH(CH.sub.2).sub.11CH.sub.3 ##STR01338## ##STR01339## 1836
10a 338 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01340## ##STR01341##
1963 10b 339 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01342##
##STR01343## 1863 10b 340 NHCO(CH.sub.2).sub.12CH.sub.3
##STR01344## ##STR01345## 2006 10b 341
NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01346## ##STR01347## 1805 10b
342 NHCO(CH.sub.2).sub.9CH.sub.3 ##STR01348## ##STR01349## 1773 10b
343 NHCO(CH.sub.2).sub.10CH.sub.3 ##STR01350## ##STR01351## 1786
10b 344 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01352## ##STR01353##
1814 10b 345 NHCO(CH.sub.2).sub.12CH.sub.3 ##STR01354##
##STR01355## 1756 10a 346 NHCO(CH.sub.2).sub.12CH.sub.3
##STR01356## ##STR01357## 1836 10a 347 NHCO(CH.sub.2).sub.7CH.sub.3
##STR01358## ##STR01359## 1765 10a 348 NHCO(CH.sub.2).sub.7CH.sub.3
##STR01360## ##STR01361## 1686 10a 349 NHCO(CH.sub.2).sub.7CH.sub.3
##STR01362## ##STR01363## 1792 10a 350 ##STR01364## ##STR01365##
##STR01366## 1832 10b 351 NHCO(CH.sub.2).sub.11CH.sub.3
##STR01367## ##STR01368## 1801 10b 352
NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01369## ##STR01370## 1801 10b
355 NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01371## ##STR01372## 1743
10a 356 NHCONH(CH.sub.2).sub.10CH.sub.3 ##STR01373## ##STR01374##
1822 10a 358 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01375## ##STR01376##
1893 1 359 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01377## ##STR01378##
948 1 360 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01379## ##STR01380##
938 1 361 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01381## ##STR01382##
952 1 362 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01383## ##STR01384##
969 1 363 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01385## ##STR01386##
970 1 364 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01387## ##STR01388##
976 1 365 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01389## ##STR01390##
976 1 366 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01391## ##STR01392##
984 1 367 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01393## ##STR01394##
984 1 368 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01395## ##STR01396##
986 1 369 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01397## ##STR01398##
987 1 370 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01399## ##STR01400##
978 1 371 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01401## ##STR01402##
998 1 372 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01403## ##STR01404##
1003 1 373 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01405## ##STR01406##
1003 1 374 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01407## ##STR01408##
970 1 375 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01409## ##STR01410##
950 1 376 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01411## ##STR01412##
950 1
377 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01413## ##STR01414## 950 1
378 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01415## ##STR01416## 955 1
379 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01417## ##STR01418## 957 1
380 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01419## ##STR01420## 958 1
381 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01421## ##STR01422## 959 1
382 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01423## ##STR01424## 959 1
383 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01425## ##STR01426## 965 1
384 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01427## ##STR01428## 965 1
385 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01429## ##STR01430## 975 1
386 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01431## ##STR01432## 975 1
387 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01433## ##STR01434## 975 1
388 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01435## ##STR01436## 957 1
389 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01437## ##STR01438## 976 1
390 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01439## ##STR01440## 976 1
391 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01441## ##STR01442## 976 1
392 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01443## ##STR01444## 983 1
393 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01445## ##STR01446## 983 1
394 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01447## ##STR01448## 948 1
395 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01449## ##STR01450## 941 1
398 NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01451## ##STR01452## 1 399
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01453## ##STR01454## 1 400
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01455## ##STR01456## 1 401
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01457## ##STR01458## 1 402
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01459## ##STR01460## 1 403
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01461## ##STR01462## 1 404
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01463## ##STR01464## 1 405
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01465## ##STR01466## 1 406
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01467## ##STR01468## 1 407
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01469## ##STR01470## 1 408
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01471## ##STR01472## 1 409
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01473## ##STR01474## 1 410
NHCO(CH.sub.2).sub.8CH.sub.3 ##STR01475## ##STR01476## 1
[0142] Preferred compounds of the present invention are compounds
45, 54, 76, 81, 85, 102, 209, 212, 253, 260, 262, 282, 285, 319,
322, 333, 334, 335, 336, 344 and 355.
[0143] According to a preferred embodiment, the present invention
provides one or more crystalline forms of compounds of formula (I)
and salts thereof
Lipopeptide Intermediates
[0144] The present invention also provides compounds that are
particularly useful as intermediates for the preparation of the
compounds of Formula I. These compounds may also have antibacterial
properties, as discussed above. In one aspect of the invention,
compounds of Formula II are provided:
##STR01477##
wherein R.sup.14 is selected from the group consisting of
##STR01478##
wherein R.sup.56 is an optionally substituted straight-chain
C.sub.8-C.sub.14 alkyl group and wherein q' is 0-3.
[0145] In another aspect of the invention, compounds of Formula III
are provided as useful intermediates for the preparation of
compounds of Formula I and/or as antibacterial compounds:
##STR01479##
wherein R.sup.15 is selected from hydrido and a carbamate amino
protecting group, preferably a tert-butoxycarbonyl group; wherein
R.sup.16 is selected from the group consisting of
##STR01480##
wherein R.sup.57 is a halo or halo substituted alkyl group,
preferably a fluoro or trifluoromethyl group; wherein, R.sup.20 is
an amino acid side chain, preferably a lysine or tryptophan side
chain.
[0146] Compounds 2, 10, 25, 38, 45, 50, 54, 76, 78, 79, 80, 81, 82,
84, 85, 103, 105, 107, 111, 115, 130, 138, 139, 146, 147, 150, 158,
164, 168, 174, 210, 212, 227, 253, 274, 275, 280, 283, 285, 317,
372 and 386 are useful both as antibacterial compounds and as
intermediates in the synthesis of compounds of this invention.
Lipopeptide Compound Pharmaceutical Compositions and Methods of Use
Thereof
[0147] Another object of the instant invention is to provide
lipopeptide compounds or salts thereof, as well as pharmaceutical
compositions or formulations comprising lipopeptide compounds or
its salts.
[0148] Lipopeptide compounds, or pharmaceutically acceptable salts
thereof, can be formulated for oral, intravenous, intramuscular,
subcutaneous or parenteral administration for the therapeutic or
prophylactic treatment of diseases, particularly bacterial
infections. For oral or parenteral administration, lipopeptide
compounds of this invention can be mixed with conventional
pharmaceutical carriers and excipients and used in the form of
tablets, capsules, elixirs, suspensions, syrups, wafers and the
like. The compositions comprising a compound of this invention will
contain from about 0.1 to about 99% by weight of the active
compound, and more generally from about 10 to about 30%.
[0149] The pharmaceutical preparations disclosed herein are
prepared in accordance with standard procedures and are
administered at dosages that are selected to reduce, prevent or
eliminate the infection (See, e.g., Remington's Pharmaceutical
Sciences, Mack Publishing Company, Easton, Pa. and Goodman and
Gilman's The Pharmaceutical Basis of Therapeutics, Pergamon Press,
New York, N.Y., the contents of which are incorporated herein by
reference, for a general description of the methods for
administering various antimicrobial agents for human therapy). The
compositions of the invention (preferably of Formula I) can be
delivered using controlled (e.g., capsules) or sustained release
delivery systems (e.g., bioerodable matrices). Exemplary delayed
release delivery systems for drug delivery that are suitable for
administration of the compositions of the invention (preferably of
Formula I) are described in U.S. Pat. Nos. 4,452,775 (issued to
Kent), 5,239,660 (issued to Leonard), 3,854,480 (issued to
Zaffaroni).
[0150] The pharmaceutically-acceptable compositions of the present
invention comprise one or more compounds of the invention
(preferably compounds of Formula I) in association with one or more
nontoxic, pharmaceutically-acceptable carriers and/or diluents
and/or adjuvants and/or excipients, collectively referred to herein
as "carrier" materials, and if desired other active ingredients.
The compositions may contain common carriers and excipients, such
as corn starch or gelatin, lactose, sucrose, microcrystalline
cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride
and alginic acid. The compositions may contain croscarmellose
sodium, microcrystalline cellulose, corn starch, sodium starch
glycolate and alginic acid.
[0151] Tablet binders that can be included are acacia,
methylcellulose, sodium carboxymethylcellulose,
polyvinylpyrrolidone (Povidone), hydroxypropyl methylcellulose,
sucrose, starch and ethylcellulose.
[0152] Lubricants that can be used include magnesium stearate or
other metallic stearates, stearic acid, silicone fluid, talc,
waxes, oils and colloidal silica.
[0153] Flavoring agents such as peppermint, oil of wintergreen,
cherry flavoring or the like can also be used. It may also be
desirable to add a coloring agent to make the dosage form more
aesthetic in appearance or to help identify the product.
[0154] For oral use, solid formulations such as tablets and
capsules are particularly useful. Sustained release or enterically
coated preparations may also be devised. For pediatric and
geriatric applications, suspensions, syrups and chewable tablets
are especially suitable. For oral administration, the
pharmaceutical compositions are in the form of, for example, a
tablet, capsule, suspension or liquid. The pharmaceutical
composition is preferably made in the form of a dosage unit
containing a therapeutically-effective amount of the active
ingredient. Examples of such dosage units are tablets and capsules.
For therapeutic purposes, the tablets and capsules which can
contain, in addition to the active ingredient, conventional
carriers such as binding agents, for example, acacia gum, gelatin,
polyvinylpyrrolidone, sorbitol, or tragacanth; fillers, for
example, calcium phosphate, glycine, lactose, maize-starch,
sorbitol, or sucrose; lubricants, for example, magnesium stearate,
polyethylene glycol, silica, or talc; disintegrants, for example,
potato starch, flavoring or coloring agents, or acceptable wetting
agents. Oral liquid preparations generally are in the form of
aqueous or oily solutions, suspensions, emulsions, syrups or
elixirs may contain conventional additives such as suspending
agents, emulsifying agents, non-aqueous agents, preservatives,
coloring agents and flavoring agents. Examples of additives for
liquid preparations include acacia, almond oil, ethyl alcohol,
fractionated coconut oil, gelatin, glucose syrup, glycerin,
hydrogenated edible fats, lecithin, methyl cellulose, methyl or
propyl para-hydroxybenzoate, propylene glycol, sorbitol, or sorbic
acid.
[0155] For intravenous (IV) use, a lipopeptide compound according
to the invention can be dissolved or suspended in any of the
commonly used intravenous fluids and administered by infusion.
Intravenous fluids include, without limitation, physiological
saline or Ringer's solution. Intravenous administration may be
accomplished by using, without limitation, syringe, minipump or
intravenous line.
[0156] Formulations for parenteral administration can be in the
form of aqueous or non-aqueous isotonic sterile injection solutions
or suspensions. These solutions or suspensions can be prepared from
sterile powders or granules having one or more of the carriers
mentioned for use in the formulations for oral administration. The
compounds can be dissolved in polyethylene glycol, propylene
glycol, ethanol, corn oil, benzyl alcohol, sodium chloride, and/or
various buffers.
[0157] For intramuscular preparations, a sterile formulation of a
lipopeptide compound or a suitable soluble salt form of the
compound, for example the hydrochloride salt, can be dissolved and
administered in a pharmaceutical diluent
such as Water-for-Injection (WFI), physiological saline or 5%
glucose. A suitable insoluble form of the compound may be prepared
and administered as a suspension in an aqueous base or a
pharmaceutically acceptable oil base, e.g., an ester of a long
chain fatty acid such as ethyl oleate.
[0158] A dose of an intravenous, intramuscular or parental
formulation of a lipopeptide compound may be adminstered as a bolus
or by slow infusion. A bolus is a dose that is administered in less
than 30 minutes. In a preferred embodiment, a bolus is administered
in less than 15 or less than 10 minutes. In a more preferred
embodiment, a bolus is administered in less than 5 minutes. In an
even more preferred embodiment, a bolus is administered in one
minute or less. An infusion is a dose that is administered at a
rate of 30 minutes or greater. In a preferred embodiment, the
infusion is one hour or greater. In another embodiment, the
infusion is substantially constant.
[0159] For topical use the compounds of the present invention can
also be prepared in suitable forms to be applied to the skin, or
mucus membranes of the nose and throat, and can take the form of
creams, ointments, liquid sprays or inhalants, lozenges, or throat
paints. Such topical formulations further can include chemical
compounds such as dimethylsulfoxide (DMSO) to facilitate surface
penetration of the active ingredient.
[0160] For application to the eyes or ears, the compounds of the
present invention can be presented in liquid or semi-liquid form
formulated in hydrophobic or hydrophilic bases as ointments,
creams, lotions, paints or powders.
[0161] For rectal administration the compounds of the present
invention can be administered in the form of suppositories admixed
with conventional carriers such as cocoa butter, wax or other
glyceride.
[0162] Alternatively, the compounds of the present invention can be
in powder form for reconstitution in the appropriate
pharmaceutically acceptable carrier at the time of delivery. In
another embodiment, the unit dosage form of the compound can be a
solution of the compound or preferably a salt thereof in a suitable
diluent in sterile, hermetically sealed ampoules or sterile
syringes. The concentration of the compound in the unit dosage may
vary, e.g. from about 1 percent to about 50 percent, depending on
the compound used and its solubility and the dose desired by the
physician. If the compositions contain dosage units, each dosage
unit preferably contains from 1-500 mg of the active material. For
adult human treatment, the dosage employed preferably ranges from 5
mg to 10 g, per day, depending on the route and frequency of
administration.
[0163] In another aspect, the invention provides a method for
inhibiting the growth of microorganisms, preferably bacteria,
comprising contacting said organisms with a compound of the
invention, preferably a compound of Formula I, under conditions
which permit entry of the compound into said organism and into said
microorganism. Such conditions are known to one skilled in the art
and are exemplified in the Examples. This method involves
contacting a microbial cell with a therapeutically-effective amount
of compound(s) of the invention, preferably compound(s) of Formula
I, in vivo or in vitro.
[0164] According to this aspect of the invention, the novel
compositions disclosed herein are placed in a pharmaceutically
acceptable carrier and are delivered to a recipient subject
(preferably a human) in accordance with known methods of drug
delivery. In general, the methods of the invention for delivering
the compositions of the invention in vivo utilize art-recognized
protocols for delivering the agent with the only substantial
procedural modification being the substitution of the compounds of
the invention (preferably compounds of Formula I) for the drugs in
the art-recognized protocols. Likewise, the methods for using the
claimed composition for treating cells in culture, for example, to
eliminate or reduce the level of bacterial contamination of a cell
culture, utilize art-recognized protocols for treating cell
cultures with antibacterial agent(s) with the only substantial
procedural modification being the substitution of the compounds of
the invention (preferably compounds of Formula I) for the agents
used in the art-recognized protocols.
[0165] In one embodiment, the invention provides a method for
treating an infection, especially those caused by gram-positive
bacteria, in a subject with a therapeutically-effective amount of a
lipopeptide compound according to Formula I. Exemplary procedures
for delivering an antibacterial agent are described in U.S. Pat.
No. 5,041,567, issued to Rogers and in PCT patent application
number EP94/02552 (publication no. WO 95/05384), the entire
contents of which documents are incorporated in their entirety
herein by reference. As used herein the phrase
"therapeutically-effective amount" means an amount of a compound of
the present invention that prevents the onset, alleviates the
symptoms, or stops the progression of a bacterial infection. The
term "treating" is defined as administering, to a subject, a
therapeutically-effective amount of a compound of the invention
(preferably a compound of Formula I) both to prevent the occurrence
of an infection and to control or eliminate an infection. The term
"subject", as described herein, is defined as a mammal, a plant or
a cell culture. In a preferred embodiment, a subject is a human or
other animal patient in need of lipopeptide compound treatment.
[0166] The method comprises administering to the subject an
effective dose of a compound of this invention. An effective dose
is generally between about 0.1 and about 100 mg/kg of a lipopeptide
compound of Formula I or a pharmaceutically acceptable salt
thereof. A preferred dose is from about 0.1 to about 50 mg/kg of a
lipopeptide compound of Formula I or a pharmaceutically acceptable
salt thereof. A more preferred dose is from about 1 to 25 mg/kg of
a lipopeptide compound of Formula I or a pharmaceutically
acceptable salt thereof. An effective dose for cell culture is
usually between 0.1 and 1000 .mu.g/mL, more preferably between 0.1
and 200 .mu.g/mL.
[0167] The compound of Formula I can be administered as a single
daily dose or in multiple doses per day. The treatment regime may
require administration over extended periods of time, e.g., for
several days or for from two to four weeks. The amount per
administered dose or the total amount administered will depend on
such factors as the nature and severity of the infection, the age
and general health of the patient, the tolerance of the patient to
the compound and the microorganism or microorganisms involved in
the infection. A method of administration to a patient of
daptomycin, another member of the lipopeptide compound class, is
disclosed in U.S. Ser. No. 09/406,568, filed Sep. 24, 1999, which
claims the benefit of U.S. Provisional Application Nos. 60/101,828,
filed Sep. 25, 1998, and 60/125,750, filed Mar. 24, 1999.
[0168] A lipopeptide compound according to this invention may also
be administered in the diet or feed of a patient or animal. If
administered as part of a total dietary intake, the amount of
compound employed can be less than 1% by weight of the diet and
preferably no more than 0.5% by weight. The diet for animals can be
normal foodstuffs to which the compound can be added or it can be
added to a premix.
[0169] The methods of the present invention comprise administering
a lipopeptide compound of Formula I or a pharmaceutical composition
thereof to a subject in need thereof in an amount that is
efficacious in reducing or eliminating the bacterial infection. The
compound may be administered orally, parenterally, by inhalation,
topically, rectally, nasally, buccally, vaginally, or by an
implanted reservoir, external pump or catheter. The compound may be
prepared for opthalmic or aerosolized uses. The compounds of the
present invention can be administered as an aerosol for the
treatment of pneumonia or other lung-based infections. A preferred
aerosol delivery vehicle is an anhydrous or dry powder inhaler.
Lipopeptide compounds of Formula I or a pharmaceutical composition
thereof also may be directly injected or administered into an
abscess, ventricle or joint. Parenteral administration includes
subcutaneous, intravenous, intramuscular, intra-articular,
intra-synovial, cisternal, intrathecal, intrahepatic, intralesional
and intracranial injection or infusion. In a preferred embodiment,
lipopeptide compounds are administered intravenously,
subcutaneously or orally. In a preferred embodiment for
administering a lipopeptide compound according to Formula I to a
cell culture, the compound may be administered in a nutrient
medium.
[0170] The method of the instant invention may be used to treat a
subject having a bacterial infection in which the infection is
caused or exacerbated by any type of bacteria, particularly
gram-positive bacteria. In one embodiment, a lipopeptide compound
or a pharmaceutical composition thereof is administered to a
patient according to the methods of this invention. In a preferred
embodiment, the bacterial infection may be caused or exacerbated by
gram-positive bacteria. These gram-positive bacteria include, but
are not limited to, methicillin-susceptible and
methicillin-resistant staphylococci (including Staphylococcus
aureus, S. epidermidis, S. haemolyticus, S. hominis, S.
saprophyticus, and coagulase-negative staphylococci), glycopeptide
intermediary-susceptible S. areus (GISA), penicillin-susceptible
and penicillin-resistant streptococci (including Streptococcus
pneumoniae, S. pyogenes, S. agalactiae, S. avium, S. bovis, S.
lactis, S. sangius and Streptococci Group C, Streptococci Group G
and viridans streptococci), enterococci (including
vancomycin-susceptible and vancomycin-resistant strains such as
Enterococcus faecalis and E. faecium), Clostridium difficile, C.
clostridiiforme, C. innocuum, C. perfringens, C. ramosum,
Haemophilus influenzae, Listeria monocytogenes, Corynebacterium
jeikeium, Bifidobacterium spp., Eubacterium aerofaciens, E. lentum,
Lactobacillus acidophilus, L. casei, L. plantarum, Lactococcus
spp., Leuconostoc spp., Pediococcus, Peptostreptococcus anaerobius,
P. asaccarolyticus, P. magnus, P. micros, P. prevotii, P.
productus, Propionibacterium acnes, Actinomyces spp., Moraxella
spp. (including M. catarrhalis) and Escherichia spp. (including E.
coli).
[0171] In a preferred embodiment, the antibacterial activity of
lipopeptide compounds of Formula I against classically "resistant"
strains is comparable to that against classically "susceptible"
strains in in vitro experiments. In another preferred embodiment,
the minimum inhibitory concentration (MIC) value for lipopeptide
compounds according to this invention against susceptible strains
is typically the same or lower than that of vancomycin. Thus, in a
preferred embodiment, a lipopeptide compound of this invention or a
pharmaceutical composition thereof is administered according to the
methods of this invention to a patient who exhibits a bacterial
infection that is resistant to other compounds, including
vancomycin or daptomycin. In addition, unlike glycopeptide
antibiotics, lipopeptide compounds exhibits rapid,
concentration-dependent bactericidal activity against gram-positive
organisms. Thus, in a preferred embodiment, a lipopeptide compound
according to this invention or a pharmaceutical composition thereof
is administered according to the methods of this invention to a
patient in need of rapidly acting antibiotic therapy.
[0172] The method of the instant invention may be used for any
bacterial infection of any organ or tissue in the body. In a
preferred embodiment, the bacterial infection is caused by
gram-positive bacteria. These organs or tissue include, without
limitation, skeletal muscle, skin, bloodstream, kidneys, heart,
lung and bone. The method of the invention may be used to treat,
without limitation, skin and soft tissue infections, bacteremia and
urinary tract infections. The method of the invention may be used
to treat community acquired respiratory infections, including,
without limitation, otitis media, sinusitis, chronic bronchitis and
pneumonia, including pneumonia caused by drug-resistant S.
pneumoniae or H. influenzae. The method of the invention also may
be used to treat mixed infections that comprise different types of
gram-positive bacteria, or which comprise both gram-positive and
gram-negative bacteria. These types of infections include
intra-abdominal infections and obstetrical/gynecological
infections. The method of the invention also may be used to treat
an infection including, without limitation, endocarditis,
nephritis, septic arthritis, intra-abdominal sepsis, bone and joint
infections and osteomyelitis. In a preferred embodiment, any of the
above-described diseases may be treated using lipopeptide compounds
according to this invention or pharmaceutical compositions
thereof.
[0173] The method of the instant invention may also be practiced
while concurrently administering one or more other antimicrobial
agents, such as antibacterial agents (antibiotics) or antifungal
agents. In one aspect, the method may be practiced by administering
more than one lipopeptide compounds according to this invention. In
another embodiment, the method may be practiced by administering a
lipopeptide compound according to this invention with another
lipopeptide compound, such as daptomycin.
[0174] Antibacterial agents and classes thereof that may be
co-administered with a compound of the present invention include,
without limitation, penicillins and related drugs, carbapenems,
cephalosporins and related drugs, aminoglycosides, bacitracin,
gramicidin, mupirocin, chloramphenicol, thiamphenicol, fusidate
sodium, lincomycin, clindamycin, macrolides, novobiocin,
polymyxins, rifamycins, spectinomycin, tetracyclines, vancomycin,
teicoplanin, streptogramins, anti-folate agents including
sulfonamides, trimethoprim and its combinations and pyrimethamine,
synthetic antibacterials including nitrofurans, methenamine
mandelate and methenamine hippurate, nitroimidazoles, quinolones,
fluoroquinolones, isoniazid, ethambutol, pyrazinamide,
para-aminosalicylic acid (PAS), cycloserine, capreomycin,
ethionamide, prothionamide, thiacetazone, viomycin, eveminomycin,
glycopeptide, glycylcylcline, ketolides, oxazolidinone; imipenen,
amikacin, netilmicin, fosfomycin, gentamicin, ceftriaxone, Ziracin,
LY 333328, CL 331002, HMR 3647, Linezolid, Synercid, Aztreonam, and
Metronidazole, Epiroprim, OCA-983, GV-143253, Sanfetrinem sodium,
CS-834, Biapenem, A-99058.1, A-165600, A-179796, KA 159, Dynemicin
A, DX8739, DU 6681; Cefluprenam, ER 35786, Cefoselis, Sanfetrinem
celexetil, HGP-31, Cefpirome, HMR-3647, RU-59863, Mersacidin, KP
736, Rifalazil; Kosan, AM 1732, MEN 10700, Lenapenem, BO 2502A,
NE-1530, PR 39, K130, OPC 20000, OPC 2045, Veneprim, PD 138312, PD
140248, CP 111905, Sulopenem, ritipenam acoxyl, RO-65-5788,
Cyclothialidine, Sch-40832, SEP-132613, micacocidin A, SB-275833,
SR-15402, SUN A0026, TOC 39, carumonam, Cefozopran, Cefetamet
pivoxil, and T 3811.
[0175] In a preferred embodiment, antibacterial agents that may be
co-administered with a compound according to this invention
include, without limitation, imipenen, amikacin, netilmicin,
fosfomycin, gentamicin, ceftriaxone, teicoplanin, Ziracin, LY
333328, CL 331002, HMR 3647, Linezolid, Synercid, Aztreonam, and
Metronidazole.
[0176] Antifungal agents that may be co-administered with a
compound according to this invention include, without limitation,
Caspofungen, Voriconazole, Sertaconazole, IB-367, FK-463,
LY-303366, Sch-56592, Sitafloxacin, DB-289 polyenes, such as
Amphotericin, Nystatin, Primaricin; azoles, such as Fluconazole,
Itraconazole, and Ketoconazole; allylamines, such as Naftifine and
Terbinafine; and anti-metabolites such as Flucytosine. Other
antifungal agents include without limitation, those disclosed in
Fostel et al., Drug Discovery Today 5:25-32 (2000), herein
incorporated by reference. Fostel et al. disclose antifungal
compounds including Corynecandin, Mer-WF3010, Fusacandins,
Artrichitin/LL 15G256, Sordarins, Cispentacin, Azoxybacillin,
Aureobasidin and Khafrefungin.
[0177] Lipopeptide compounds may be administered according to this
method until the bacterial infection is eradicated or reduced. In
one embodiment, a lipopeptide compound is administered for a period
of time from 3 days to 6 months. In a preferred embodiment, a
lipopeptide compound is administered for 7 to 56 days. In a more
preferred embodiment, a lipopeptide compound is administered for 7
to 28 days. In an even more preferred embodiment, a lipopeptide
compound is administered for 7 to 14 days. Lipopeptide compounds
may be administered for a longer or shorter time period if it is so
desired.
General Procedures for Lipopeptide Compound Synthesis
[0178] Lipopeptide compounds of Formula I may be produced as
described below. The lipopeptide compounds of the instant invention
may be produced semi-synthetically using daptomycin as a starting
point or may be produced by a total synthesis approach.
[0179] For the semi-synthetic approach according to the present
invention, daptomycin may be prepared by any method known in the
art. See, e.g., U.S. Pat. Nos. 4,885,243 and 4,874,843. Daptomycin
may be used in its acylated state or it may be deacylated prior to
its use as described herein. Daptomycin may be deacylated using
Actinoplanes utahensis as described in U.S. Pat. No. 4,482,487.
Alternatively, daptomycin may be deacylated as follows:
[0180] Daptomycin (5.0 g) was dissolved in water (25 ml) and
adjusted to pH 9 with 5M sodium hydroxide. Ditert-butyldicarbonate
(1.5 g) was added and the mixture was adjusted to maintain pH 9
with 5 M sodium hydroxide until the reaction was complete (4
hours). The pH was adjusted to 7 and the mixture was loaded onto a
Bondesil 40.mu. C8 resin column. The column was washed with water
and the product was eluted from the column with methanol.
Evaporation of the methanol gave BOC-protected daptomycin as a
yellow powder.
[0181] A preparation of deacylase enzyme was produced from
recombinant Streptomyces lividans, which expresses the Actinoplanes
utahensis deacylase enzyme. The enzyme in ethylene glycol (400
.mu.l) was added to BOC-protected daptomycin (1 g) in water (100
ml) at pH 7-8. After incubation for 72 hours, the mixture was
loaded on a Bondesil 40.mu. C8 resin column. The column was washed
with water and the product was eluted from the column with 10%
acetonitrile in water. The product was evaporated to give
deacylated BOC-protected daptomycin as a yellow powder.
Kynurenine Derivatives
##STR01481##
[0183] Daptomycin can be converted into analogs bearing
modifications at the R.sup.2 position by converting the aromatic
amino group to the diazonium salt compound I with reagents such as
sodium nitrite/hydrochloric acid or isoamylnitrite. Using chemistry
known to those skilled in the art and following the teachings of
the disclosure, the diazonium group can then be displaced by
reagents such as sodium azide, potassium ethylxanthate or copper
chloride to yield derivative compounds II, wherein R.sup.19 is as
previously defined.
##STR01482##
[0184] Additionally, compound I can be converted to the azide
compound III by reaction with an azide source, typically sodium
azide. Modifications to the ketone group can then be undertaken
using chemistry known to those having ordinary skill in the art,
such as reduction, oxime formation, ketalization conversion to a
leaving group and displacement to give compounds of formula IV,
wherein R.sup.17 and R.sup.18 are as previously defined.
##STR01483##
[0185] Compound IV may also be converted to compound V by reducing
the azide group to the amine using chemistry known to those having
ordinary skill in the art, and following the teachings of the
disclosure, such as reaction with triphenyl phosphine and water, or
reducing agents such as sodium borohydride wherein R.sup.17 and
R.sup.18 are as previously defined.
##STR01484##
[0186] Additionally compound I can be converted into compound VI by
reduction with hypophosphorus acid. Modifications to the ketone
group can then be undertaken using chemistry known to those having
ordinary skill in the art similar to those used in scheme 2,
wherein R.sup.17 and R.sup.18 are as previously defined.
Ornithine Derivatives
##STR01485##
[0188] Daptomycin can be converted into analogs bearing
modifications at the R.sup.1 position by treating the aromatic
amino group of the ornithine with reagents such as isocyanates,
isothiocyanates, activated esters, acid chlorides,
sulfonylchlorides or activated sulfonamides, heterocycles bearing
readily displaceable groups, imidates, lactones or reductively with
aldehydes to yield compound VIII, wherein R.sup.1 is as previously
defined.
Tryptophan Amine Derivatives
##STR01486##
[0190] Daptomycin can be converted into compound IX by first
protecting the ornithine amine with an appropriate amino protecting
group (P) known to those skilled in the art and following the
teachings of the disclosure. The decyl side chain on the tryptophan
is then removed using an enzyme capable of deacylating daptomycin,
such as that described above.
##STR01487##
[0191] Compound IX can be modified at the tryptophan amine with
reagents such as isocyanates, isothiocyanates, activated esters,
acid chlorides, sulfonylchlorides or activated sulfonamides,
heterocycles bearing readily displaceable groups, imidates,
lactones or reductively with aldehydes to yield compound X.
Compound X can be deprotected to give compound XI according to
procedures known to those skilled in the art following the
disclosure of this invention, wherein R is as previously
defined.
[0192] The above modifications to the ornithine amine R.sup.1,
tryptophan amine R or kynurenine side chain R.sup.2 may be
independently combined to yield additional compounds that are
modified at up to all three sites. In order to achieve these
modifications, it may be necessary to protect certain
functionalities in the molecule. Protecting these functionalities
should be within the expertise of one skilled in the art following
the disclosure of this invention. See, e.g., Greene, supra.
Solid Support Synthesis of Lipopeptide Compounds
[0193] In an alternative embodiment of the invention, the
lipopeptide compounds of Formula I may be synthesized on a solid
support as outlined below. In step 1, a
suitably-N-protected-.beta.MeGlu(OH)--OAllyl ester is coupled to a
suitable resin to give Compound XII. Deprotection of the amino
group of Compound XII, followed by coupling of the amino group with
a suitably protected seryl derivative (A1) gives Compound XIII,
wherein P is a suitable protecting group. This peptide coupling
process, i.e., deprotection of the alpha-amino group, followed by
coupling to a suitably protected amino acid, is repeated until the
desired number of amino acids have been coupled to the resin. In
the scheme shown below, eleven amino acids have been coupled to
give Compound XIV. Addition of an activated R group, R*, is added
to Compound XIV to give Compound XV. In step 4, Compound XV is
cyclized to give Compound XVI. Subsequently, in step 5, Compound
XVI is removed from the resin to give the lipopeptide Compound
XVII.
##STR01488##
##STR01489##
wherein A.sup.1, is a suitably protected serine derivative, wherein
R.sup.31 is a suitable, cleavable hydroxyl protecting group as
outlined below.
##STR01490##
wherein A.sup.2 and A.sup.7, are suitably protected glycine
derivatives as outlined below.
##STR01491##
wherein A.sup.3, A.sup.5 and A.sup.9 are suitably protected
aspartic acid derivatives as outlined below, wherein .sup.28R,
.sup.29R and .sup.30R are cleavable protecting groups, preferably
t-butyl groups.
##STR01492##
wherein A.sup.4 is a suitably protected alanine derivative as
outlined below.
##STR01493##
wherein A.sup.6 is a suitably protected ornithine derivative as
outlined below, or derivatized ornthine wherein *R.sup.1 is R.sup.1
as previously described or alternatively a protected form of
R.sup.1 that would yield R.sup.1 upon subsequent deprotection.
##STR01494##
wherein A.sup.8 is a suitably protected depsipeptide as outlined
below, Y is a protecting group that is cleavable under conditions
that leave other protecting groups intact to the others used, i.e.,
Alloc; and wherein *R.sup.2 is R.sup.2 as previously described or
alternatively a protected form of R.sup.2 that would yield R.sup.2
upon subsequent deprotection. Preferably .sup.2*R is a kynurenine,
or substituted kynurenine side chain, most preferably
##STR01495##
wherein A.sup.10 is a suitably protected asparagine derivative as
outlined below.
##STR01496##
wherein A.sup.11 is a suitably protected tryptophan derivative as
outlined below, wherein R*.sup.37 is hydrido or a suitable
protecting group, preferably t-butoxy carbonyl.
[0194] It will be understood by those skilled in the art that both
the amino and the side chain functional groups must be suitably
protected prior to attaching them to the growing peptide chain.
Suitable protecting groups can be any group known in the art to be
useful in peptide synthesis. Such pairings of protecting groups are
well known. See, e.g., "Synthesis Notes" in the Novabiochem Catalog
and Peptide Synthesis Handbook (1999), pages S1-S93 and references
cited therein. Following the disclosure of the present application,
the selection of protecting groups and method of use thereof will
be known to one skilled in the art.
[0195] It will also be understood by those skilled in the art that
the choice of protecting group on the side chain functional groups
will either result or not result in the protecting group being
cleaved concomitantly with the peptide's final cleavage from the
resin, which will give the natural amino acid functionality or a
protected derivative thereof, respectively.
[0196] The following general procedures serve to exemplify the
solid support synthesis of compounds of Formula I.
Step 1: Coupling Suitably-N-Protected-.beta.MeGlu(OH)--OAllyl Ester
to a Resin
[0197] Five molar equivalents each, with respect to the resin, of a
suitably-N-protected-.beta.MeGlu(OH)--OAllyl ester,
1,3-Diisopropylcarbodiimide (DIC) and 1-Hydroxy-7-azabenzotriazole
(HOAt) are stirred for 30 mins in dimethylformamide (DMF; 5 ml/g
resin). A suitably functionalised resin or solid support, such as,
but not limited to, Wang, Safety Catch, Rink, Knorr, PAL, or PAM
resin, is added and the resulting suspension is stirred for 16 hrs.
The resin-N-protected-.beta.MeGlu(OH)--OAllyl ester is then
filtered, dried and the coupling is repeated. The N-protecting
group is then removed using the appropriate conditions given in the
coupling steps below.
Step 2: (A) General Coupling Cycle for Amino Acids with an
N-9-Fluorenylmethoxycarbonyl (Fmoc) Protecting Group
[0198] Five molar equivalents each, with respect to the resin-AA
(wherein resin-AA is defined as the resin attached the growing
amino acid chain), of a suitably protected Fmoc amino acid, DIC,
and HOAt (0.5 molar solution in DMF) are added to the resin-AA,
along with sufficient DMF to give a working volume. The mixture is
shaken for one hour, filtered, and the coupling is repeated. After
the second coupling the resin is washed twice with DMF, twice with
methanol, and twice again with DMF. The Fmoc group of the newly
coupled amino acid A.sup.1-11 is deprotected by stirring the resin
product in one working volume of a solution of 20% piperidine in
N-methylpyrrolidine for five minutes, filtering the resin, and
stirring the resin in 20% piperidine in N-methylpyrrolidine again
for 20 minutes. The resin is washed twice with DMF, twice with
methanol, and twice again with DMF.
Step 2 (B): General Coupling Cycle of Amino Acids with an
N-tert-Butoxy-carbonyl (N-Boc) Protecting Group
[0199] Five molar equivalents each, with respect to the resin-AA,
of a suitably protected N-Boc amino acid, DIC, and HOAt (0.5 molar
solution in DMF) are added to the resin-AA, along with sufficient
DMF to give a working volume. The mixture is shaken for one hour,
filtered, and the coupling is repeated. After the repeated coupling
the resin is washed twice with DMF, twice with methanol, and twice
again with DMF. The Boc group of the newly coupled amino acid
A.sup.1-11, is then deprotected by stirring the resin in one
working volume of CH.sub.2Cl.sub.2:trifluoroacetic acid (TFA) 1:1
for 15 minutes, filtering, and stirring in one working volume of
CH.sub.2Cl.sub.2:TFA 1:1 for another 15 minutes. The resin is
neutralized by washing with excess diisopropylethylamine (DIPEA) in
CH.sub.2Cl.sub.2 and then washed twice with DMF, twice with
methanol, and twice again with DMF.
Step 3: Terminal Amine Capping Reaction
[0200] Ten molar equivalents, with respect to the resin XV, of a
suitable reagent containing R* such as an activated ester,
isocyanate, thioisocyanate, anhydride, acid chloride,
chloroformate, or reactive salt thereof, in one working volume of
DMF is added to the resin XIV and agitated for 25 hours. The
resulting resin XV is washed twice with DMF, twice with methanol,
and twice again with DMF.
Step 4: Cyclization
[0201] The dried resin XV is placed under an argon atmosphere, and
treated with a solution of Pd(PPh.sub.3).sub.4 125 mgs/0.1 mmol
peptide substrate, in CH.sub.2Cl.sub.2:Acetic
acid:N-Methylmorpholine, 40:2:1, 1 ml/0.1 mmol peptide substrate.
The mixture is stirred for 3 hours at ambient temperature,
filtered, and washed twice with DMF, twice with methanol, and twice
again with DMF. Five molar equivalents each, with respect to the
resin, of DIC, and HOAt (0.5 molar solution in DMF) are added to
the resin, along with sufficient DMF to give a working volume. The
reaction is shaken for 17 hours, filtered, and washed twice with
DMF, twice with methanol, and twice again with DMF to give resin
XVI.
Step 5: Cleavage and Isolation of the Lipopeptide
[0202] The desired lipopeptide is cleaved from resin XVI and
isolated, resulting in a compound in which R.sup.27 is OH or
NH.sub.2. If Fmoc chemistry is used, the dried resin is suspended
in 1 ml/0.1 mmol peptide substrate of
CH.sub.2Cl.sub.2:TFA:Ethanedithiol (EDT):Triisopropylsilane (TIS),
16:22:1:1, and stirred for 6-8 hours at ambient temperature. The
resin is filtered, washed with 1 equal volume of cold TFA, and the
combined filtrates are evaporated under reduced pressure. Crude
product XVII is then precipitated by the addition of diethyl ether,
and isolated by centrifugation. This product may be further
purified by preparative reverse phase HPLC.
[0203] If N-Boc chemistry is used, the dried resin is suspended in
hydrogen fluoride (HF):anisole:dimethylsulfide (DMS), 10:1:1, and
stirred for 2 hours at 0.degree. C. The volitiles are evaporated
under a stream of nitrogen. The resin is then extracted with TFA,
filtered and washed twice with TFA, and the combined TFA filtrates
evaporated under reduced pressure. Crude product is then
precipitated by the addition of diethyl ether, and isolated by
centrifugation. This product may be further purified by preparative
reverse phase HPLC.
[0204] If the resin is a Safety Catch resin, then R.sup.27.dbd.OR
or NRH. The dried resin XVI is suspended in N-methylpyrrolidine
(NMP) or dimethylsulphoxide (DMSO) (8 ml/g resin), Five equivalents
of DIPEA (with respect to resin substitution) and 24 equivalents of
iodo or bromoacetonitrile (with respect to resin substitution) are
added. The suspension is stirred for 24 hours at ambient
temperature under inert atmosphere. The resin is filtered, washed
with tetrahydrofuran (THF) and DMSO. For an ester, the resin is
then treated with an alcohol, hydroxide or alkoxide (20 equivalents
with respect to resin substitution) in THF for 20 hours. The resin
is filtered, washed with THF and water, and the combined filtrates
are evaporated under reduced pressure. Crude product is
precipitated by the addition of diethyl ether, and isolated by
centrifugation. The product may be further purified by preparative
reverse phase HPLC. For amides the resin is then treated with a
primary or secondary amine (20 equivalents with respect to resin
substitution) in THF for 12-40 hours, at a gentle reflux under
inert atmosphere. The resin is filtered, washed with THF and water,
and the combined filtrates are evaporated under reduced pressure.
Crude product is then precipitated by the addition of diethyl
ether, and isolated by centrifugation. This product may be further
purified by preparative reverse phase HPLC.
[0205] In order that this invention may be more fully understood,
the following examples are set forth. These examples are for the
purpose of illustration only and are not to be construed as
limiting the scope of the invention in any way.
EXAMPLE 1
Preparation of Compounds 38, 40, 50, 52, 77-80, 82-84, 87-100,
103-169, 171-176, 183-187, 194-199, 201-204, 208, 210-211, 222-244,
252, 265-267, 271-281, 283-284, 286-291, 323-331, 358-395 and
398-410
[0206] A suspension of daptomycin in dry dimethylformamide (0.6 ml)
was treated with a solution of 4-Fluorobenzaldehyde (0.2 ml) and a
suspension of sodium triacetoxyborohydride (0.2 ml, 1.5M in dry
dimethylformamide). After 24 hours, the reaction mixture was
diluted with water/acetonitrile (1:1; 0.4 ml) and purified by
preparative HPLC. The reaction mixture was loaded onto an IBSIL-C8
5.mu. 250.times.20.2 mm column and eluted at 20 ml/min with 30-60%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing product were collected and freeze-dried. The
freeze-dried residue was dissolved in water (5 ml) and applied to a
Bondesil 40.mu. C8 resin column. The column was washed with water
and eluted with methanol. Evaporation of the methanol gave compound
38 as a pale yellow solid (23 mg).
[0207] In an analogous manner, compounds 40, 50, 52, 77-80, 82-84,
87-100, 103-169, 171-176, 183-187, 194-199, 201-204, 208, 210-211,
222-244, 252, 265-267, 271-281, 283-284, 286-291, 323-331, 358-395
and 398-410 can be prepared as detailed in the above example by
appropriate substitutions of reagents obvious to those skilled in
the art following the teachings of the disclosure.
EXAMPLE 1a
Preparation of Compound 282
[0208] 2-Methyl-6-nitroquinoline (0.4 ml, 0.5M solution in dioxane)
was treated with selenium dioxide (0.2 ml, 0.9M solution in 9/1
dioxane/water) and heated to 90.degree. C. overnight. The mixture
was cooled to room temperature and diluted with water (1 ml). The
mixture was then extracted with ethyl acetate (3.times.2 ml). The
organic extract was then dried in vacuo to give
6-nitro-2-quinolinecarboxaldehyde which was carried forward without
further purification. Daptomycin (1 ml, 0.1 M in dry
dimethylformamide) was treated successively with
6-nitro-2-quinolinecarboxaldehyde prepared above in dry
dimethylformamide (0.2 ml) and sodium triacetoxyborohydride (0.4
ml, 1.5M solution in dry dimethylformamide). The mixture was capped
and shaken briefly. After 24 h, the mixture was treated with water
(0.2 ml) and loaded onto an IBSIL-C8 5.mu. 250.times.20.2 mm
column. The column was eluted at 25 ml/min under the gradient
conditions of 30-60% acetonitrile in 5 mM ammonium phosphate buffer
over 25 min followed by holding at 60% acetonitrile in 5 mM
ammonium phosphate buffer for another 10 min. The desired fractions
were collected and the acetonitrile was removed by evaporation. The
residue was applied to a Bondesil 40.mu. C8 resin column, washed
with water and the product was eluted with methanol. Evaporation of
the methanol gave compound 282 as a pale yellow solid.
EXAMPLE 1b
Preparation of Compound 285
[0209] 4-Chloro-2-methylquinoline (0.4 ml, 0.5M solution in
dioxane) was treated with selenium dioxide (0.2 ml, 0.9M solution
in 9/1 dioxane/water) and heated to 90.degree. C. overnight. The
mixture was cooled to room temperature and diluted with water (1
ml). The mixture was then extracted with ethyl acetate (3.times.2
ml). The organic extract was then dried in vacuo to give
4-chloro-2-quinolinecarboxaldehyde which was carried forward
without further purification. Daptomycin (1 ml, 0.1 M in dry
dimethylformamide) was treated successively with
4-chloro-2-quinolinecarboxaldehyde prepared above and diluted in
dry dimethylformamide (0.2 ml) and sodium triacetoxyborohydride
(0.4 ml, 1.5M in dry dimethylformamide). The mixture was capped and
shaken briefly. After 24 h the mixture was treated with water (0.2
ml) and loaded on an IBSIL-C8 5.mu. 250.times.20.2 mm column. The
column was eluted at 25 ml/min under the gradient conditions of
30-60% acetonitrile in 5 mM ammonium phosphate buffer over 25 min
followed by holding at 60% acetonitrile in 5 mM ammonium phosphate
buffer for another 10 min. The desired fractions were collected and
the acetonitrile was removed by evaporation. The residue was
applied to a Bondesil 40.mu. C8 resin column, washed with water and
the product eluted off with methanol. Evaporation of the methanol
gave compound 285 as a yellow solid.
EXAMPLE 1c
Preparation of Compound 85
[0210] Daptomycin (1 ml, 0.1M in dry dimethylformamide) was treated
successively with 1-methyl-2-imidazolecarboxaldehyde (0.2 ml, 0.5M
solution in dry dimethylformamide) and sodium triacetoxyborohydride
(0.4 ml, 1.5M solution in dry dimethylformamide). The mixture was
capped and shaken briefly. After 24 h the mixture was treated with
water (0.2 ml) and loaded onto an IBSIL-C8 5.mu. 250.times.20.2 mm
column. The column was eluted at 30 ml/min under the gradient
conditions of 35-40% acetonitrile in 5 mM ammonium phosphate buffer
over 30 min. The desired fractions were collected and the
acetonitrile was removed by evaporation. The residue was applied to
a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. This mixture was then loaded on a Prodigy ODS 10.mu.
250.times.21.2 mm column eluted at 50 ml/min at 33% acetonitrile in
5 mM ammonium phosphate buffer adjusted to pH 3.2. The desired
fractions were collected and the acetonitrile was removed by
evaporation. The residue was applied to a Bondesil 40.mu. C8 resin
column, washed with water and the product was eluted with methanol.
Evaporation of the methanol gave compound 85 as a pale yellow
solid.
EXAMPLE 1d
Preparation of Compound 212
[0211] Daptomycin (1 ml, 0.1 M in dry dimethylformamide) was
treated successively with 2-imidazolecarboxaldehyde (0.2 ml, 0.5M
solution in dry dimethylformamide) and sodium triacetoxyborohydride
(0.4 ml, 1.5M solution in dry dimethylformamide). The mixture was
capped and shaken briefly. After 24 h, the mixture was treated with
water (0.2 ml) and the mixture was loaded on an IBSIL-C8 5.mu.
250.times.20.2 mm column. The column was eluted at 30 ml/min under
the gradient conditions of 35-40% acetonitrile in 5 mM ammonium
phosphate buffer over 30 min. The desired fractions were collected
and the acetonitrile was removed by evaporation. The residue was
applied to a Bondesil 40.mu. C8 resin column, washed with water and
eluted with methanol. This mixture was then loaded on a Prodigy ODS
10.mu. 250.times.21.2 mm column and eluted at 50 ml/min at 33%
acetonitrile in 5 mM ammonium phosphate buffer adjusted to pH 3.2.
The desired fractions were collected and the acetonitrile was
removed by evaporation. The residue was applied to a Bondesil
40.mu. C8 resin column, washed with water and the product eluted
with methanol. Evaporation of the methanol gave compound 212 as a
yellow solid.
EXAMPLE 1e
Preparation of Compound 81
[0212] Daptomycin (1 ml, 0.1M in dry dimethylformamide) was treated
successively with 5-fluoroindole-3-carboxaldehyde (0.2 ml, 0.5M
solution in dry dimethylformamide) and sodium triacetoxyborohydride
(0.4 ml, 1.5M solution in dry dimethylformamide). The mixture was
capped and shaken briefly. After 24 h the mixture was treated with
water (0.2 ml) and loaded on an IBSIL-C8 5.mu. 250.times.20.2 mm
column. The column was eluted at 25 ml/min under the gradient
conditions of 30-60% acetonitrile in 5 mM ammonium phosphate buffer
over 25 min followed by holding at 60% acetonitrile in 5 mM
ammonium phosphate buffer for another 10 min. The desired fractions
were collected, the acetonitrile was removed by evaporation and the
residue applied to a Bondesil 40.mu. C8 resin column. The column
was washed with water and the product was eluted with methanol.
Evaporation of the methanol gave compound 81 as a pale yellow
solid.
EXAMPLE 1f
Preparation of Compound 253
[0213] p-N,N-Bis(2-chloroethyl)aminobenzaldehyde (0.3 g) was
dissolved in acetone (2.5 ml) and treated with sodium iodide (0.4
g). The mixture was warmed to 40.degree. C. for 3 h then treated
with benzylamine (0.2 ml) and triethylamine (0.4 ml). The mixture
was diluted to 7 ml with acetonitrile and then heated to 60.degree.
C. After 24 h, the mixture was cooled to room temperature and the
solvent was removed by evaporation.
4-(4-Benzylpiperazino)benzaldehyde was purified by silica gel
chromatography eluting with (10%
triethylamine/methanol/dichloromethane).
[0214] Daptomycin (1 ml, 0.1 M in dry dimethylformamide) was
treated successively with the 4-(4-benzylpiperazino)benzaldehyde
prepared above diluted in dry dimethylformamide (0.2 ml), and
sodium triacetoxyborohydride (0.4 ml, 1.5M solution in dry
dimethylformamide). The mixture was capped and shaken briefly.
After 24 h the mixture was treated with water (0.2 ml) and loaded
on an IBSIL-C8 5.mu. 250.times.20.2 mm column. The column was
eluted at 25 ml/min under the gradient conditions of 30-60%
acetonitrile in 5 mM ammonium phosphate buffer over 25 min followed
by holding at 60% acetonitrile in 5 mM ammonium phosphate buffer
for another 10 min. The desired fractions were collected, the
acetonitrile was removed by evaporation and the residue was applied
to a Bondesil 40.mu. C8 resin column. The column was washed with
water and the product was eluted with methanol. Evaporation of the
methanol gave compound 253 as a pale yellow solid.
EXAMPLE 1g
Preparation of Compound 76 and 177
[0215] Daptomycin (1 ml, 0.1 M in dry dimethylformamide) was
treated successively with 4-phenylbenzaldehyde (0.2 ml, 0.5M in dry
dimethylformamide) and sodium triacetoxyborohydride (0.4 ml, 1.5M
in dry dimethylformamide). The reaction mixture was capped and
shaken briefly to mix the solution. After 24 h the mixture was
treated with water (0.2 ml) and loaded on an IBSIL-C8 5.mu.
250.times.20.2 mm column. The column was eluted at 25 ml/min under
the gradient conditions of 30-60% acetonitrile in 5 mM ammonium
phosphate buffer over 25 min followed by holding at 60%
acetonitrile in 5 mM ammonium phosphate buffer for another 10 min.
The desired fractions were collected, the acetonitrile was removed
by evaporation and the residue was applied to a Bondesil 40.mu. C8
resin column. The column was washed with water and the product was
eluted with methanol. Evaporation of the methanol gave compound 76
as a pale yellow solid. Compound 177 was obtained by deacylation of
compound 76 according to Example 7.
EXAMPLE 1h
Preparation of Compound 209
[0216] 4-Hydroxy-3-nitrobenzaldehyde (0.4 ml, 0.2M in acetone) was
successively treated with potassium hydroxide (0.1 ml, 1M in water)
and 4-fluorobenzylbromide (0.4 ml, 0.2M in acetone). After 24 h the
mixture was dried in vacuo to give
4-(4-fluorobenzyloxy)-3-nitro-benzaldehyde which was carried
forward without further purification.
[0217] Daptomycin (1 ml, 0.1 M in dry dimethylformamide) was
treated successively with,
4-(4-fluorobenzyloxy)-3-nitro-benzaldehyde previously prepared
above diluted in dry dimethylformamide (0.2 ml), and sodium
triacetoxyborohydride (0.4 ml, 1.5M in dry dimethylformamide). The
mixture was capped and shaken briefly. After 24 h the mixture was
treated with water (0.2 ml) and loaded onto an IBSIL-C8 5.mu.
250.times.20.2 mm column. The column was eluted at 25 ml/min under
the gradient conditions of 30-60% acetonitrile in 5 mM ammonium
phosphate buffer over 25 min followed by holding at 60%
acetonitrile in 5 mM ammonium phosphate buffer for another 10 min.
The desired fractions were collected, the acetonitrile was removed
by evaporation and the residue was applied to a Bondesil 40.mu. C8
resin column. The column was washed with water and the product was
eluted with methanol. Evaporation of the methanol gave compound 209
as a pale yellow solid.
EXAMPLE 2
Preparation of Compounds 10, 11-17, 19-20, 22-27 and 190
[0218] Daptomycin (972 mg) was dissolved in dry dimethylformamide
(20 ml), and isatoic anhydride (979 mg) was added. The mixture was
stirred at ambient temperature for 10 days, then quenched by the
addition of water (20 ml). The mixture was loaded onto a Bondesil
40.mu. C8 resin column (25 g), which had been previously washed
with methanol (50 ml) and water (100 ml). The column was then
eluted with water (200 ml), 15% methanol/water (1200 ml), 20%
methanol/water (200 ml), 30% methanol/water (200 ml) and 40%
methanol/water (200 ml). The product bearing fractions were
combined and freeze dried to give compound 10 as a white solid (870
mg).
[0219] In an analogous manner, compounds 11-17, 19-20, 22-27 and
190 can be prepared as detailed in the above example by appropriate
substitutions of reagents obvious to those skilled in the art
following the teachings of the disclosure.
EXAMPLE 3
Preparation of Compounds 44, 45, 41-43, 46-48, 55-58, 60-75,
178-180, 193 and 245
[0220] Daptomycin (500 mg) and Boc-tryptophan-p-nitrophenyl ester
(157.5 mg) were stirred at room temperature in dry
dimethylformamide (30 ml) for 3 days. Water (30 ml) was added and
the mixture was purified on a Bondesil 40.mu. C8 resin column (25
g). The column was eluted with 20% acetonitrile in water (200 ml),
40% acetonitrile in water (200 ml) and finally with methanol.
Evaporation of the solvent from the product-containing fractions
gave compound 44 as a pale yellow solid (450 mg).
[0221] Compound 44 (200 mg) was cooled to 0.degree. C. and a
0.degree. C. solution of 5% thioanisole in trifluoroacetic acid (10
ml) was added. After 3 hours at 0.degree. C. the mixture was
evaporated to dryness and the residue was purified by preparative
HPLC on an IBSIL-C8 5.mu. 250.times.20.2 mm column. The column was
eluted at 20 ml/min with 38% acetonitrile in 5 mM ammonium
phosphate buffer. The product containing fractions were
freeze-dried. The freeze-dried residue was dissolved in water (5
ml) and applied to a Bondesil 40.mu. C8 resin column. The column
was washed with water and eluted with methanol. Evaporation of the
methanol gave compound 45 as a pale yellow solid.
[0222] In an analogous manner, compounds 41-43, 46-48, 55-58,
60-75, 178-180, 193 and 245 can be prepared as detailed in the
above example by appropriate substitutions of reagents obvious to
those skilled in the art following the teachings of the
disclosure.
EXAMPLE 3a
Preparation of Compounds 54, 49 and 51
[0223] Daptomycin (400 mg) and
N,N-bis(tert-butoxycarbonyl)-L-lysine-4-nitrophenyl ester (173 mg)
were stirred in dry dimethylformamide (5 ml) at room temperature
for two days. The mixture was loaded onto an IBSIL-C8 5.mu.
250.times.20.2 mm column and was eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave the Boc protected
intermediate as a pale yellow solid (370 mg).
[0224] Boc protected intermediate (200 mg) was stirred in
trifluoroacetic acid (5 ml) and anisole (0.25 ml) at room
temperature for 2 hours. Removal of the solvents under reduced
pressure gave a residue which was loaded on an IBSIL-C8 5.mu.
250.times.20.2 mm column and eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave compound 54 as a
pale yellow solid (100 mg).
[0225] In an analogous manner, compounds 49 and 51 can be prepared
as detailed in the above example by appropriate substitutions of
reagents obvious to those skilled in the art following the
teachings of the disclosure.
EXAMPLE 3
Preparation of Compounds 32, 18, 21, 28-31, 33-35, 39, 182 and
189
[0226] Daptomycin (162 mg) and 2-methylthiobenzoic acid
pentafluorophenol ester (37 mg) were stirred at room temperature in
dry dimethylformamide (10 ml) for 5 days. The dimethylformamide was
evaporated under reduced pressure and the residue was purified by
preparative HPLC on an IBS-C8 5.mu. 250.times.20.2 mm column. The
column was eluted at 20 ml/min with 36% acetonitrile in 5 mM
ammonium phosphate buffer. Fractions collected at 7.3 minutes were
freeze-dried. The freeze-dried residue was dissolved in water (5
ml) and applied to a Bondesil 40.mu. C8 resin column. The column
was washed with water and eluted with methanol. Evaporation of the
methanol gave compound 32 as a pale yellow solid (47 mg).
[0227] In an analogous manner, compounds 18, 21, 28-31, 33-35, 39,
182 and 189 can be prepared as detailed in the above example by
appropriate substitutions of reagents by one having ordinary skill
in the art following the teachings of the disclosure.
EXAMPLE 4
Preparation of Compounds 5, 4, 6-8 and 9
[0228] Daptomycin (16 mg) was dissolved in dry dimethylformamide
(0.5 ml) and methyl isothiocyanate (37 mg) was added. The mixture
was stirred at ambient temperature for 24 hours, then quenched by
the addition of 5% ammonium phosphate buffer (1 ml). The mixture
was purified by preparative HPLC on an IBSIL-C8 5.mu.
250.times.20.2 mm column. The column was eluted at 20 ml/min with
36% acetonitrile in 5 mM ammonium phosphate buffer. The product
bearing fractions were combined and freeze dried. The freeze-dried
residue was dissolved in water (1.5 ml) and applied to a Bondesil
40.mu. C8 resin column. The column was washed with water and eluted
with methanol. Evaporation of the methanol gave compound 5 as a
pale yellow solid (5.2 mg).
[0229] In an analogous manner, compounds 4, 6-8 and 9 can be
prepared as detailed in the above example by appropriate
substitutions of reagents obvious to those having ordinary skill in
the art.
EXAMPLE 5
Preparation of Compound 3
[0230] Daptomycin (16 mg) and N-benzotriazole phenylsulfonamide
(2.6 mg) were stirred at room temperature in dry pyridine for 6
days. The solvent was evaporated and the residue was purified by
preparative HPLC using an IBSIL-C8 5.mu. 250.times.20.2 mm column.
The column was eluted at 20 ml/min with 36% acetonitrile in 5 mM
ammonium phosphate buffer and product containing fractions were
freeze-dried. The freeze dried residue was dissolved in water (5
ml) and applied to a Bondesil 40.mu. C8 resin column. The column
was washed with water and eluted with methanol. Evaporation of the
methanol gave compound 3 as a pale yellow solid (4 mg).
EXAMPLE 6
Preparation of Compounds 1, 2, 221, 259 and 270
[0231] Daptomycin (32 mg) was dissolved in dry dimethylformamide
(20 ml), and N,N'-bis-Boc-1-guanidinylpyrazole (31 mg) was added.
The mixture was stirred at ambient temperature for 5 days, then
quenched by the addition of water (3 ml). The resultant mixture was
loaded onto a Bondesil 40.mu. C8 resin (900 mg) that had been
previously washed with methanol and water. The column was eluted
with water (30 ml) followed by methanol. The product-bearing
fractions were combined and evaporated to give compound 1 as a
white solid.
[0232] Compound 1 (30 mg) was dissolved in trifluoroacetic
acid/dichloromethane/tri-isopropylsilane/ethane dithiol
(11/8/0.5/0.5, 3 ml) and stirred at ambient temperature for 90
minutes. The mixture was evaporated to dryness and the residue was
precipitated by the addition of diethyl ether (10 ml). The residue
was purified by preparative HPLC on an IBSIL-C8 5.mu.
250.times.20.2 mm column. The column was eluted at 20 ml/min with
38% acetonitrile in 5 mM ammonium phosphate buffer. The
product-bearing fractions were combined and freeze dried. The
freeze-dried residue was dissolved in water (1.5 ml) and applied to
a Bondesil 40.mu. C8 resin column. The column was washed with water
and eluted with methanol. Evaporation of the methanol gave compound
2 as a white solid (6.4 mg).
[0233] In an analogous manner, compounds 221, 259 and 270 can be
prepared as detailed in the above example by appropriate
substitutions of reagents obvious to those having ordinary skill in
the art following the teachings of the disclosure.
EXAMPLE 7
Preparation of Compounds 255, 260, 254, 256-257, 261, 263, 292-294
and 313-314
[0234] Daptomycin (10 g) was dissolved in dry dimethylformamide
(100 ml). N,N'-bis-Boc-guanidinylpyrazole (2.3 g) in dry
dimethylformamide (5 ml) was added. The mixture was stirred under
nitrogen at room temperature overnight. The mixture was purified on
a Bondesil 40.mu. C8 resin column. The product containing fractions
were freeze-dried to give compound 1 (7.4 g) as pale yellow fluffy
solid.
[0235] Compound 1 (2.6 g) was added to a preparation of deacylase
enzyme produced from recombinant Streptomyces lividans, which
expresses the Actinoplanes utahensis deacylase enzyme in ethylene
glycol (1.2 ml) and water (25 ml). The pH of the solution was
adjusted to 9 with 1.0M sodium hydroxide solution and stirred at
room temperature. After 24 hours the mixture was purified on a
Bondesil 40.mu. C8 resin column by eluting with 10%
acetonitrile/water, then 40% acetonitrile/water. The
product-containing fractions were freeze dried to give deacylated
bis-Boc-guanidinylated daptomycin (0.69 g) as a pale yellow
solid.
[0236] Undecanoyl pentafluorophenol ester (40.3 mg) was added to
deacylated bis-Boc-guanidinylated daptomycin (171.5 mg) in dry
dimethylformamide (2 ml). The mixture was stirred overnight at room
temperature before being concentrated to give compound 255 (105 mg)
as a yellow solid.
[0237] Compound 255 was dissolved in trifluoroacetic acid (5.5 ml),
dichloromethane (4 ml), ethane dithiol (0.25 ml) and
triisopropylsilane (0.25 ml). The mixture was stirred for 4 hours
at room temperature before being concentrated and purified by
preparative HPLC on an IB-SIL 5.mu. 250.times.20.2 mm column. The
column was eluted at 25 ml/min with acetonitrile and ammonium
phosphate buffer 30%-60% gradient for 40 min. The desired fractions
were collected at 21 minutes and freeze dried. The freeze-dried
residue was dissolved in water and applied to a Bondesil C8 resin
column. The column was washed with water and eluted with methanol.
Evaporation of the methanol gave compound 260 (27.8 mg) as a pale
yellow solid.
[0238] In an analogous manner, compounds 254, 256-257, 261, 263,
292-294 and 313-314 can be prepared as detailed in the above
example by appropriate substitutions of reagents obvious to those
having ordinary skill in the art following the disclosure of the
invention.
EXAMPLE 7a
Preparation of Compounds 258 and 262
[0239] Tetradecanoyl pentafluorophenol ester (35.5 mg) and
deacylated bis-Boc-guanidinylated daptomycin (102.5 mg) in dry
dimethylformamide (2 ml). The mixture was stirred overnight at room
temperature before being concentrated to give compound 258 (38.8
mg) as a yellow solid.
[0240] Compound 258 (38.8 mg) was dissolved in trifluoroacetic acid
(5.5 ml), dichloromethane (4 ml), ethane dithiol (0.25 ml) and
triisopropylsilane (0.25 ml). The mixture was stirred for 4 hours
at room temperature before being concentrated and purified by
preparative HPLC on an IB-SIL 5.mu. 250.times.20.2 mm column. The
column was eluted at 25 ml/min with acetonitrile and ammonium
phosphate buffer 30%-60% gradient for 40 min. The desired fractions
were collected at 21 minutes and freeze dried. The freeze-dried
residue was dissolved in water and applied to a Bondesil C8 resin
column. The column was washed with water and eluted with methanol.
Evaporation of the methanol gave compound 262 (2.1 mg) as a pale
yellow solid.
EXAMPLE 8
Preparation of Compound 37, 36 and 192
[0241] Daptomycin (162 mg) was stirred in 0.1 M hydrochloric acid
(5 ml) at 0.degree. C. for 10 minutes before sodium nitrite (8 mg)
in water (0.2 ml) was added dropwise. Sulfamic acid (11 mg) was
added after 15 minutes, followed by sodium azide (8 mg) 10 minutes
later. The mixture was maintained at 0.degree. C. for 4 hours and
then neutralized with a saturated sodium bicarbonate solution and
purified by preparative HPLC. An IBSIL-C8 5.mu. 250.times.20.2 mm
column was loaded with the mixture and eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions were
collected at 6.9 minutes and freeze dried. The freeze-dried residue
was dissolved in water (5 ml) and applied to a Bondesil 40.mu. C8
resin column. The column was washed with water and eluted with
methanol. Evaporation of the methanol gave the azido daptomycin as
a pale yellow solid (60 mg).
[0242] The azido daptomycin (69 mg) was dissolved in dry
dimethylformamide (4 ml) and iminobiotin-N-hydroxysuccinimide ester
(53 mg) was added. The mixture was covered to exclude light and
stirred at ambient temperature for 3 days. The mixture was quenched
by the addition of water (20 ml). The resultant mixture was loaded
onto a Bondesil 40.mu. C8 resin (25 g) column, which had been
previously washed with methanol and water, and the column was
eluted with water. The product-bearing fractions were combined and
freeze dried to give Compound 37 as a white solid (49 mg).
[0243] In an analogous manner, compounds 36 and 192 can be prepared
as detailed in the above example by appropriate substitutions of
reagents obvious to those having ordinary skill in the art by
following the disclosure of the invention.
EXAMPLE 8
Preparation of Compound 200
[0244] Daptomycin (1.62 g) in 50% wt aqueous solution of
hypophosphorus acid (10 ml) was stirred at 0.degree. C. for 30
minutes before adding dropwise a solution of sodium nitrite (76 mg)
in water (0.5 ml). The mixture was allowed to come to room
temperature and stirred for 24 hours. The mixture was purified by
preparative HPLC by loading the mixture on an IBSIL-C8 5.mu.
250.times.20.2 mm column and eluting the column at 20 ml/min with
32% acetonitrile in 5 mM ammonium phosphate buffer. The desired
fractions were collected at 30 minutes and freeze dried. The
freeze-dried residue was dissolved in water (5 ml) and applied to a
Bondesil 40.mu. C8 resin column. The column was washed with water
and eluted with methanol. Evaporation of the methanol gave desamino
daptomycin as a pale yellow solid (200 mg).
[0245] To desamino daptomycin (80 mg) in dry dimethylformamide (2
ml) was added N-t-butoxycarbonyl-L-tryptophan-p-nitrophenyl ester
(32 mg). The mixture was stirred at room temperature for 24 hours
before being purified by preparative HPLC. The mixture was loaded
on an IBSIL-C8 5.mu. 250.times.20.2 mm column and eluted at 20
ml/min with 40% acetonitrile in 5 mM ammonium phosphate buffer. The
desired fractions were collected at 19 minutes and freeze-dried.
The freeze-dried residue was dissolved in water (2 ml) and applied
to a plug of Bondesil 40.mu. C8 resin (500 mg). The Bondesil resin
was washed with water (10 ml) and then the product was eluted with
methanol (10 ml). Evaporation of the methanol gave Boc protected
compound 200 as a pale yellow solid (20 mg).
[0246] To Boc protected compound 200 (20 mg) in 60% trifluoroacetic
acid in dichloromethane (0.5 ml) was added anisole (10 .mu.L). The
mixture was stirred at room temperature for 6 hours before being
evaporated to dryness. Preparative HPLC purification of the residue
was done on an IBSIL-C8 5.mu. 250.times.20.2 mm column and eluted
at 20 ml/min with 38% acetonitrile in 5 mM ammonium phosphate
buffer. The desired fractions were collected at 15 minutes and
freeze-dried. The freeze-dried residue was dissolved in water (2
ml) and applied to a plug of Bondesil 40.mu. C8 resin (500 mg). The
Bondesil resin was washed with water (10 ml) and the product was
eluted with methanol (10 ml). Evaporation of the methanol gave
compound 200 as a pale yellow solid (4 mg).
EXAMPLE 9
Preparation of Compounds 181, 86, 101-102, 206-207, 213-220,
246-251, 264 and 269
[0247] Daptomycin (250 mg) and N-tBoc-L-tryptophan-p-nitrophenyl
ester (144 mg) were stirred in dry dimethylformamide (3 ml) at room
temperature for two days. The mixture was loaded on an IBSIL-C8
5.mu. 250.times.20.2 mm column and was eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave N-Boc tryptophan
daptomycin as a pale yellow solid (130 mg).
[0248] A preparation of deacylase enzyme was produced from
recombinant Streptomyces lividans, which expresses the Actinoplanes
utahensis deacylase enzyme. The enzyme in ethylene glycol (400
.mu.l) was added to the solution of N-Boc tryptophan daptomycin
(100 mg) in HPLC grade water (20 ml). The solution was adjusted to
pH 8.5 with sodium hydroxide (1 M). The mixture was stirred for 24
hours. The mixture was loaded on a C8 resin plug column, washed
with water and eluted with methanol. Evaporation of the methanol
gave a residue which was applied to an IBSIL-C8 5.mu.
250.times.20.2 mm column and was eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave deacylated N-Boc
tryptophan daptomycin as a pale yellow solid (42 mg).
[0249] Deacylated N-Boc tryptophan daptomycin (20 mg) was stirred
in dry dimethylformamide (2 ml) at room temperature. Undecyl
isocyanate (2.25 mg) was added to the solution. After stirring at
ambient temperature for 24 hours, the mixture was diluted with
water (10 ml) and applied to a Bondesil 40.mu. C8 resin column,
washed with water and eluted with methanol. Evaporation of the
methanol gave the undecyl urea of N-Boc tryptophan daptomycin as a
pale yellow solid (21 mg).
[0250] N-Boc tryptophan daptomycin undecyl urea (21 mg) was stirred
in trifluoroacetic acid (2 ml) and anisole (0.1 ml) at room
temperature for 2 hours. Removal of the solvents under reduced
pressure gave a residue which was loaded on an IBSIL-C8 5.mu.
250.times.20.2 mm column and eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave compound 181 as a
pale yellow solid (0.8 mg).
[0251] In an analogous manner, compounds 86, 101-102, 206-207,
213-220, 246-251, 264 and 269 can be prepared as detailed in the
above example by appropriate substitutions of reagents obvious to
those having ordinary skill in the art following the disclosure of
the invention.
EXAMPLE 9
Preparation of Compound 205
[0252] Deacylated N-Boc tryptophan daptomycin (50 mg) and
nonaldehyde (4.1 mg) were stirred in dry dimethylformamide (2 ml)
at room temperature. Sodium triacetoxy borohydride (3.6 mg) was
added to the solution. The mixture was stirred for 24 hours, then
loaded on an IBSIL-C8 5.mu. 250.times.20.2 mm column and eluted at
20 ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.
Fractions containing the desired compound were collected and
freeze-dried. The freeze-dried residue was dissolved in water (5
ml) and applied to a Bondesil 40.mu. C8 resin column, washed with
water and eluted with methanol. Evaporation of the methanol gave
nonyl amino N-Boc tryptophan daptomycin as a pale yellow solid (14
mg).
[0253] Nonyl amino N-Boc tryptophan daptomycin (14 mg) was stirred
in trifluoroacetic acid (2 ml) and anisole (0.1 ml) at room
temperature for 2 hours. Removal of the solvents under reduced
pressure gave a residue which was loaded on an IBSIL-C8 5.mu.
250.times.20.2 mm column and was eluted at 20 ml/min with 37%
acetonitrile in 5 mM ammonium phosphate buffer. Fractions
containing the desired compound were collected and freeze-dried.
The freeze-dried residue was dissolved in water (5 ml) and applied
to a Bondesil 40.mu. C8 resin column, washed with water and eluted
with methanol. Evaporation of the methanol gave compound 7 as a
pale yellow solid (5 mg).
EXAMPLE 10
Preparation of Compounds 356, 315-322, 332-337, 345-349 and 355
[0254] Daptomycin (5.0 g) was dissolved in water (25 ml) and
adjusted to pH 9 with 5M sodium hydroxide. Di-tert-butyldicarbonate
(1.5 g) was added and the mixture was adjusted to maintain pH 9
with 5 M sodium hydroxide until the reaction was complete (4
hours). The pH was adjusted to 7 and the mixture was loaded onto a
Bondesil 40.mu. C8 resin column. The column was washed with water
and the product was eluted from the column with methanol.
Evaporation of the methanol gave Boc-protected daptomycin (5.08 g)
as a yellow powder.
[0255] A preparation of deacylase enzyme was produced from
recombinant Streptomyces lividans, which expresses the Actinoplanes
utahensis deacylase enzyme. The enzyme in ethylene glycol (400
.mu.l) was added to Boc-protected daptomycin (1 g) in water (100
ml) at pH 7-8. After incubation for 72 hours, the mixture was
loaded on a Bondesil 40.mu. C8 resin column. The column was washed
with water and the product was eluted from the column with 10%
acetonitrile in water. The solvent was removed by evaporation to
give deacylated Boc-protected daptomycin (440 mg) as a yellow
powder.
[0256] Daptomycin undecyl urea synthesized from deacylated Boc
protected daptomycin above using undecyl isocyanate instead of
undecanoyl pentafluorophenol ester according to example 7 (100 mg)
and 5-methoxyindole-3-carboxaldehyde (11 mg) in dry
dimethylformamide (0.6 ml) was added sodium triacetoxyborohydride
(76 mg). The mixture was stirred at room temperature for 24 hours
before purification by preparative HPLC. The mixture was loaded on
an IBSIL-C8 5.mu. 250.times.20.2 mm column and eluted at 25 ml/min
with 30-60% acetonitrile in 5 mM ammonium phosphate gradient over
30 minutes. The desired fractions were collected at 21 minutes and
freeze-dried. The freeze-dried residue was dissolved in water (2
ml) and applied to a plug of Bondesil 40.mu. C8 resin (500 mg). The
Bondesil resin was washed with water (10 ml) and then the product
was eluted with methanol (10 ml). Evaporation of the methanol gave
compound 114 as a pale yellow solid (10 mg).
[0257] In an analogous manner, compounds 315-322, 332-337, 345-349
and 355 can be prepared as detailed in the above example by
appropriate substitutions of reagents obvious to those skilled in
the art.
EXAMPLE 10a
Preparation of Compounds 307, 310, 295-306, 308-309, 311-312,
338-344 and 350-352
[0258] Daptomycin undecanoyl amide synthesized from deacylated Boc
protected daptomycin by using undecanoyl pentafluorophenol ester
according to examples 10 and 7 (60 mg) was stirred in dry
dimethylformamide (2 ml) at room temperature.
N-tBoc-L-tryptophan-p-nitrophenyl ester (31 mg) was added to the
solution. The mixture was stirred for 24 hours. The mixture was
loaded onto an IBSIL-C8 5.mu. 250.times.20.2 mm column and was
eluted at 20 ml/min with 37% acetonitrile in 5 mM ammonium
phosphate buffer. Fractions containing the desired compound were
collected and freeze-dried. The freeze-dried residue was dissolved
in water (5 ml) and applied to a Bondesil 40.mu. C8 resin column,
washed with water and eluted with methanol. Evaporation of the
methanol gave compound 307 as a pale yellow solid (25 mg).
[0259] Compound 307 (20 mg) was stirred in trifluoroacetic acid (2
ml) and anisole (0.1 ml) at room temperature for 2 hours. Removal
of the solvents under reduced pressure gave a residue which was
loaded on an IBSIL-C8 5.mu. 250.times.20.2 mm column and was eluted
at 20 ml/min with 37% acetonitrile in 5 mM ammonium phosphate
buffer. Fractions containing the desired compound were collected
and freeze-dried. The freeze-dried residue was dissolved in water
(5 ml) and applied to a Bondesil 40.mu. C8 resin column, washed
with water and eluted with methanol. Evaporation of the methanol
gave compound 310 as a pale yellow solid (4 mg).
[0260] In an analogous manner, compounds 295-306, 308-309, 311-312,
338-344 and 350-352 can be prepared as detailed in the above
example by appropriate substitutions of reagents obvious to those
skilled in the art.
EXAMPLE 11
[0261] Compounds according to Formula I were tested for
antimicrobial activity against a panel of organisms according to
standard procedures described by the National Committee for
Clinical Laboratory Standards (NCCLS document M7-A5, Vol. 20, No.
2, 2000) except that all testing was performed at 37.degree. C.
Compounds were dissolved in 100% dimethyl sulfoxide and were
diluted to the final reaction concentration (0.1 .mu.g/mL-100
.mu.g/mL) in microbial growth media. In all cases the final
concentration of dimethyl sulfoxide incubated with cells is less
than or equal to 1%. For minimum inhibitory concentration (MIC)
calculations, 2-fold dilutions of compounds were added to wells of
a microtiter plate containing 5.times.10.sup.4 bacteria cells in a
final volume of 100 .mu.L of media (Mueller-Hinton Broth
supplemented with 50 mg/L Ca.sup.2+). The optical densities (OD) of
the bacterial cells, which measures bacterial cell growth and
proliferation, were measured using a commercial plate reader. The
MIC value is defined as the lowest compound concentration
inhibiting growth of the test organism. The MIC (in .mu.g/ml)
values of representative compounds of the present invention are
listed in Table III.
EXAMPLE 12
[0262] The mouse protection test is an industry standard for
measuring the efficacy of a test compound in vivo [for examples of
this model see J. J. Clement, et al., Antimicrobial Agents and
Chemotherapy, 38 (5), 1071-1078, (1994)]. As exemplified below,
this test is used to demonstrate the in vivo efficacy of the
compounds of the present invention against bacteria.
[0263] The in vivo antibacterial activity was established by
infecting female CD-1 mice (Charles River Lab, MA) weighing 19-23 g
intraperitoneally with from Methicillin Resistant S. aureus (MRSA)
inoculum. The inoculum was prepared from Methicillin Resistant S.
areus (ATCC 43300). The MRSA inoculum was cultured in
Mueller-Hinton (MH) broth at 37.degree. C. for 18 hours. The
optical density at 600 nm (OD.sub.600) was determined for a 1:10
dilution of the overnight culture. Bacteria (8.times.10.sup.8 cfu)
was added to 20 ml of phosphate buffered saline (Sigma P-0261)
containing 5% hog gastric mucin (Sigma M-2378). All animals were
injected with 0.5 ml of the inoculum, equivalent to
2.times.10.sup.7 cfu/mouse, which is the dose causing 100% death of
the animals without treatment.
[0264] The test compound was dissolved in 10.0 ml of 50 mM
phosphate buffer to give a solution of 1 mg/ml (pH=7.0). This
solution was serially diluted with vehicle by 4-fold (1.5 ml to 6.0
ml) to give 0.25, 0.063 and 0.016 mg/ml solutions. All the
solutions were filtered with 0.2 m Nalgene syringe filter.
Immediately after the bacterial inoculation, group 1 animals were
subcutaneously (sc) injected with buffer (no test compound) and
groups 2 to 5 were given test compound sc at 10.0, 2.5, 0.63, and
0.16 mg/kg, respectively. Group 6 animals received test compound sc
at 10 mg/kg (or the highest therapeutic dose of a given compound)
only for monitoring acute toxicity. These injections were repeated
once at 4 hours after the inoculation for the respective groups.
The injection volume at each time was 10 ml per kilogram of body
weight. The results of the in vivo efficacy test are summarized in
Table II, which provides a representative example of the results
obtained for Compound 70. The 50% effective dose (ED.sub.50) is
calculated on the basis of the number of mice surviving 7 days
after inoculation. The ED.sub.50 was determined for other compounds
of this invention in a similar manner. The ED.sub.50 in mg/kg of
other representative compounds of the present invention are listed
in Table III.
TABLE-US-00003 TABLE II # of Survival (7 Group mice Inoculated with
Treatment days) 1 5 MRSA #43300 Phosphate buffer 0/5 2 .times.
10.sup.7 cfu/mouse 10 ml/kg, s.c. x2 2 5 MRSA #43300 Compound 70
5/5 2 .times. 10.sup.7 cfu/mouse 10 mg/kg, s.c. x2 3 5 MRSA #43300
Compound 70 3/5 2 .times. 10.sup.7 cfu/mouse 2.5 mg/kg, s.c. x2 4 5
MRSA #43300 Compound 70 1/5 2 .times. 10.sup.7 cfu/mouse 0.63
mg/kg, s.c. x2 5 5 MRSA #43300 Compound 70 0/5 2 .times. 10.sup.7
cfu/mouse 0.16 mg/kg, s.c. x2 6 5 No Compound 70 5/5 10 mg/kg, s.c.
x2 The ED.sub.50 of compound 70 is calculated to be 1.51 mg/kg
TABLE-US-00004 TABLE III MIC MIC ED.sub.50 (.mu.g/ml) (.mu.g/ml)
mg/kg Compound # S. aureus E. faecalis S. aureus 1 ++ + ++ 2 +++ +
+++ 3 ++ + 4 + + 5 ++ ++ 6 ++ ++ 7 ++ ++ 8 ++ ++ 9 +++ ++ 10 +++ +
++ 11 ++ + 12 +++ ++ 13 +++ ++ 14 ++ ++ 15 ++ ++ 16 +++ ++ 17 ++ ++
18 ++ + 19 ++ ++ 20 +++ ++ 21 ++ + 22 ++ ++ 23 +++ ++ 24 +++ ++ ++
25 +++ ++ 26 +++ ++ 27 ++ + 28 ++ + 29 + 30 ++ + 31 ++ + 32 ++ + 33
++ + 34 ++ + 35 ++ + 36 ++ + 37 ++ + 38 +++ + 39 + + 40 ++ + 41 + +
42 ++ + 43 ++ + 44 ++ ++ 45 +++ ++ +++ 46 ++ ++ 47 ++ ++ 48 +++ ++
49 ++ ++ 50 ++ + 51 ++ ++ 52 +++ + 53 ++ + 54 ++ ++ ++ 55 +++ + 56
+++ ++ 57 ++ + 58 +++ + 60 ++ + 61 ++ + 62 ++ + 63 ++ + 64 ++ + 65
++ + 66 ++ + 67 ++ + 68 ++ + 69 ++ + 70 +++ + ++ 71 ++ + 72 ++ + 73
++ + 74 ++ 75 ++ + 76 +++ ++ ++ 77 ++ ++ 78 + + 79 +++ ++ 80 +++ ++
81 +++ ++ +++ 82 +++ ++ 83 +++ ++ 84 +++ ++ 85 +++ ++ +++ 86 + + 87
+++ ++ 88 ++ + 89 +++ ++ 90 ++ ++ 91 ++ + 92 ++ + 93 ++ ++ 94 +++
++ 95 +++ ++ 96 +++ ++ 97 +++ ++ 98 +++ ++ 99 +++ ++ 100 +++ ++ 101
++ ++ 102 +++ +++ 103 +++ + 104 ++ ++ 105 +++ ++ 106 +++ ++ 107 ++
++ 108 ++ ++ 109 ++ ++ 110 ++ ++ 111 +++ ++ 112 ++ + 113 ++ ++ 114
++ + 115 +++ + 116 +++ ++ 117 ++ ++ 118 ++ ++ 119 +++ ++ 120 ++ ++
121 +++ ++ 122 +++ + 123 ++ + 124 ++ + 125 ++ ++ 126 ++ ++ 127 +++
++ 128 ++ ++ 129 +++ + 130 +++ ++ 131 +++ + 132 ++ ++ 133 +++ ++
134 ++ + 135 +++ + 136 +++ ++ 137 ++ + 138 +++ + 139 +++ ++ 140 +++
++ 141 ++ + 142 +++ + 143 ++ + 144 +++ ++ 145 ++ ++ 146 +++ + 147
+++ ++ 148 ++ ++ 149 ++ + 150 +++ ++ 151 +++ ++ 152 ++ ++ 153 ++ +
154 ++ ++ 155 ++ ++ 156 +++ + 157 ++ + 158 ++ + 159 +++ + 160 ++ +
161 +++ + 162 ++ ++ 163 +++ ++ 164 +++ ++ 165 ++ ++ 166 +++ ++ 167
+++ ++ 168 +++ ++ 169 +++ + 170 ++ ++ 171 ++ ++ 172 +++ ++ 173 +++
++ 174 +++ ++ 175 ++ ++ 176 +++ ++ 177 + + 178 ++ + 179 ++ + 180 ++
++ 181 +++ +++ +++ 182 ++ + 183 +++ + 184 +++ + 185 ++ + 186 ++ +
187 +++ + 189 190 192 ++ + 193 ++ + 194 ++ + 195 ++ + 196 +++ + 197
++ + 198 ++ + 199 +++ + 200 + 201 ++ ++ 202 203 ++ + 204 +++ ++ 205
++ + 206 207 208 ++ ++ 209 +++ ++ 210 +++ ++ 211 ++ 212 +++ ++ +++
213 214 215 216 ++ + 217 218 + 219 +++ ++ 220 +++ +++ 221 + + 222
++ ++ 223 +++ ++ 224 ++ + 225 ++ + 226 ++ + 227 +++ ++ 228 +++ ++
229 +++ ++ 230 +++ +++ 231 +++ ++ 232 +++ ++ 233 ++ + 234 ++ + 235
+++ ++ 236 ++ + 237 +++ ++ 238 +++ ++ 239 +++ + 240 +++ ++ 241 ++
++ 242 ++ + 243 ++ + 244 +++ ++ 245 246 +
247 + 248 + 249 + 250 +++ + 251 ++ + 252 ++ ++ 253 +++ ++ 254 ++ +
255 +++ ++ 256 +++ +++ 257 ++ + 258 +++ ++ 259 +++ +++ 260 +++ ++
261 ++ ++ 262 ++ ++ 263 +++ ++ 264 ++ + 265 ++ ++ 266 +++ + 267 ++
+ 268 ++ ++ 269 + 270 +++ + 271 +++ +++ 272 ++ + 273 +++ ++ 274 +++
+++ 275 +++ ++ 276 +++ +++ 277 +++ +++ 278 +++ +++ 279 +++ ++ 280
+++ ++ 281 +++ ++ 282 +++ +++ +++ 283 +++ ++ 284 +++ +++ 285 +++
+++ +++ 286 +++ +++ 287 +++ +++ 288 +++ +++ 289 +++ ++ 290 ++ ++
291 +++ +++ 292 +++ ++ 293 +++ ++ 294 ++ + 295 ++ 296 297 ++ 298
299 ++ 300 ++ 301 +++ ++ 302 +++ ++ 303 +++ ++ 304 305 +++ ++ 306
+++ ++ 307 +++ ++ 308 +++ ++ 309 +++ ++ 310 +++ ++ 311 +++ 312 313
+++ 314 +++ ++ 315 ++ + 316 +++ 317 318 +++ +++ 319 +++ ++ 320 +++
++ 321 +++ ++ 322 +++ ++ 323 +++ ++ 324 ++ 325 +++ ++ 326 +++ ++
327 +++ ++ 328 ++ 329 330 +++ ++ 331 ++ 332 +++ ++ 333 ++ 334 +++
+++ 335 +++ ++ 336 +++ ++ 337 +++ +++ 338 ++ ++ 339 +++ ++ 340 + +
341 ++ ++ 342 +++ ++ 343 +++ ++ 344 +++ ++ 345 ++ +++ 346 +++ +++
347 ++ + 348 ++ + 349 ++ + 350 ++ + 351 ++ ++ 352 ++ ++ 355 ++ ++
356 +++ +++ 358 ++ ++ 359 +++ 360 +++ ++ 361 +++ 362 +++ ++ 363 +++
364 +++ ++ 365 +++ 366 +++ ++ 367 +++ ++ 368 +++ ++ 369 +++ 370 +++
++ 371 +++ ++ 372 +++ ++ 373 +++ ++ 374 +++ ++ 375 +++ 376 +++ ++
377 +++ ++ 378 +++ ++ 379 +++ ++ 380 +++ ++ 381 +++ ++ 382 +++ ++
383 +++ ++ 384 +++ ++ 385 +++ ++ 386 +++ ++ 387 +++ +++ 388 ++ ++
389 +++ ++ 390 +++ ++ 391 +++ ++ 392 +++ ++ 393 +++ ++ 394 +++ ++
395 +++ ++ Wherein "+++" indicates that the compound has an MIC
(.mu.g/ml) of 1 .mu.g/ml or less or an ED.sub.50 of 1 mg/kg or
less; "++" indicates that the compound has an MIC (.mu.g/ml) or
ED.sub.50 of greater than 1 .mu.g/ml or 1 mg/kg, respectively but
less than or equal to10 .mu.g/ml or ED.sub.50 of 10 mg/kg,
respectively; and "+" indicates that the compound has an MIC
(.mu.g/ml) of greater than 10 .mu.g/ml or an ED.sub.50 of greater
than 10 mg/kg.
[0265] All publications and patent applications cited in this
specification are herein incorporated by reference as if each
individual publication or patent application were specifically and
individually indicated to be incorporated by reference. Although
the foregoing invention has been described in some detail by way of
illustration and example for purposes of clarity of understanding,
it will be readily apparent to those of ordinary skill in the art
in light of the teachings of this invention that certain changes
and modifications may be made thereto without departing from the
spirit or scope of the appended claims.
* * * * *