Note: Descriptions are shown in the official language in which they were submitted.
2 13 6 3 2 Li
.
WO 94/01110P~/US93/048~0
-1-
5'-INDOLn~lYL OXAZOLIDINONES USEFUL AGAINST
~fYCOBAClERlUM TUBE;RCULOSIS
BACKGROUND OF THE_INVENTIC)N
1. Fleld of the Invention
5The invention is the use of hvo ~'-indolinyl ox~zolidionones (I) to treat tuber~ulosis
(TB) caused by M. nlberc~fosis.
2 escnption of the Related Art
Tuberculosis (TB) is a well known disease which is caused by M. nJberculosis.
At present v~rious agents are used to treat those ir~ected with TB. The most cornmon
of these pharmaceutical agents include isor~ia~id, rifampin, etharnbutol, p-aminosalicylic acid,
py~inamide, streptomycin, capreomycin, cycloserine, eWonamide and kanarnycin and- ~minoglycosides ~ntibiotics and mixt~res thereof.
Since the introduction of antibiotics and the ~etter health care practices in the 1950's,
~he incidence of TB had declined an ave~age of six percent per year until 1985. Since then
there has been a 16 percent per year increase in new TB cases. This increased incidence of TB
has been accompanied by outbreaks of multi-drug resistant strains of M. tubercl~losis. Because
TB has never declined in incidence in ~le developing and underdeveloped countries and the
additional c~es related to the increase in AIDS, the World Health Organization recently
established a Worlcing Group to develop new antibiotics to treat TB which are urgently needed.
Wh,ile there are phamlaceutical agents presently available to treat those infected with
TB, it is highly desirable to have a better agent because of the development of M. tu~erculoQs
strains resist~nt to current therapeutics.
US Patent 4,128,654 discloses 5-halomethylphenyl-2-oxazolidinones which are useful in
controlling fungal and bacterial diseases of plants.
US Patent 4,250,318 discloses 3-substituted phenyl-5-hydroxymethyloxazolidinoneshaving antidepressive utility.
US Reissue Patent 29,607 discloses 3-substituted phenyl-5-hydroxymethylox~olidinones
having antidepressive, tranquilizing and sedative utility.
US Patent 4,340,606 discloses 3-(p-alkylsulfonyl~phenyl-5-(hydroxymethyl or
acyloxyrnethyl)oxazolidinones having antibacterial activity in mamrnals.
Belgium Pa~ent 892,270 discloses 3-(arylalkyl, arylalkenyl or arylacetylenic
substituted)phenyl)-5-(~minomethyl)oxazolidinones which are inhibitors of monoamine oxid~se.
US Paten~ 4,461,773 discloses 3-substituted phenyl-5-hydroxymethyloxa~olidinoneswhich have antibactenal ~ctivity.
European Patent Publications 127,902 and 184,170 disclose 3-substituted phenyl-5-
amidomethylox~201idinones which have antibacterial utility.
2 1 3 6 ~ 2 `~
WO 94/01110 PCr/US93/b4850
-2-
Antimicrobial Agents and Chemotherapy 1791 (1987) discusses compounds disclosed in
Europe3n Patent Publications 127,902 and 184,170, discussed above, and compares these new
compounds with known antibiotics.
US Patent 4,705,799 discloses aminomethyloxooxazolidinyl benzene derivatives
5 including sulfides, su~xides, sul~ones and sulfonamides which possess antibacteriai activity
US Pa~ent 4,801,600 discloses 6'-indolinyloxazolidinones (where the indolinyl nitrogen
is meta to the oxazolidinone nitrogen~.
US Patents 5,036,092 and 5,039,690 disclose 6'-indolinyl oxazolidinones whereas the
compound of the present invention is a 5'-indolinyl oxazolidinone.
Diagn. Microbiol. Infec~. Dis., 14, 465-471 (1991) discloses that an oxazolidinone, ( )-
DUP-721 is active against tuberculosis.
5'-indolinyl oxazolidinones are known, see Intemational Publication No. WO90/02744,
published March 22, 1990 based on lntemational Patent Application No. PCI`/US89/03548 filed
August 22, 1989. More particularly, 3-(5'-1-hydroxyacetylindolinyl)-SB-(acetamidomethyl)-
oxazolidin-2-one more preferably termed 5-(S)-N-(1'-hydroxyacetyl-5'-indolinyl)-5-
(acetamidomethyl)-2-oxazolidinone is known, see Intemational Publication No. W090/02744,
EXAMPLES 122. The optically active from of the compound is disclosed in EXAMPLE 150.
5-(S)-N-(1'-(2-Thiophenecarbonyl3-5'-indolinyl)-5-(acetamidomethyl)-2-oxazolidinone is also
known, see EXAMPLE 146 of Intemational Publication No. W090/02744.
SUMMARY OF INVENTION
Disclosed is a method of treating humans who have tuberculosis caused by
Mycobacterium tl bercu~osis which comprises admisitration of an effective amount of a 5'-
indolinyl oxazolidinone selected from the group consisting of
5-(S)-N-(l'-hydroxyacetyl-5'-indolinyl)-5-(acetamidomethyl)-2-oxazolidinone and
5-(S)-N-(1 '-(2-thiopheneca~bonyl)-5 '-indolinyl)-5-(acetamidomethyl)-2-oxazolidinone
and pharmaceutically acceptable salts therPof.
- DLTAILED DESCRIPTION OF THE INVENTlON
5'-indolinyl oxazolidinones are known, see lntemational Publication No. WO90/02744,
the compounds of formula (XI). The 5'-indolinyl oxazolidinones contain an asymmetric center
and therefore produce two enantiomers one "S" and the other "R", either of which can be (+/d)
and the other (-tl). (~)-3-(5'-1-Hydroxyacetylindolinyl)-SB-(acetamidomethyl)oxazolidin-2-one is
known, see In~ernational Publication No. W090/02744, EXAMPLES 122. The antibacterially
~ctive form of the compound is the "S" enantiomer and is disclosed in EXAMPLE 150 ~s
obtained by resolution of a racemic mixture. The optically active form was telmed 3-(5'-1-
B5 hydroxyacetylindolinyl)-5B-(~ce~midomethyl)oxazolidin-2-one in International Publication No.
W090,~02744 However, a preferred name is 5-(S)-N-(1'-hydroxy~cetyl-5'-indolinvl)-5-
~- 213~324 ~ `
:
WO 94/01110 PCr~US93J048~0
(acetamidomethyl)-2-oxazolidinone. 5-(S)-N-( 1 '-(2-Thiophenec~onyl)-5 '-indolinyl)-5-
(ace~midomethyl)-2-oxazolidinone is also known, see EXA~LE 146 of International
Pu~lication ~o. WO90/02744.
While the racemic forms of the oxazolidinones are useful in treaIing TB, it is highly
S prefer~ble to use the active en~ntiomer rather than the racemic form. While the optically active ;
form can be obtained by lesolution of a racemic mixture as set forth in Intemational Publication
No. W090/02744, it is preferred to prepare the desired enantiomer by a stereoselective
synthesis. One method to produce 5-(S)-N-(1'-hydroxyacetyl-5'-indolinyl)-5-(acetamidomethyl)-
2-oxazolidinone is by resolution of the racemic form. Another method is by the process of
EXAMPLES 1-9. The preferred method, as discussed below, is by the process of EXA~LES
14-19. Likewise, one method to produce 5-(S)-N-(1'-(2-thiophenecarbonyl)-5'-indolinyl)-5-
- (~etamidomethyl)-2-oxazolidinone is by resolution of the racemic form. Another method is by
the process of EXAMPLES 10-13. The preferTed method, as discussed below, is by the process
of EXAMPLES 14-17 and 20.
The preferred method of malcing the optically active is by starting with the known 1-
carbo-t-butyloxy-(N-carboben~yloxy)-5-aminoindoline [US Patent 5,164,510, EXA~LE 11,
compound (IV)) in a solution of tetrahydrofuran or ether at -78 under an inert atmosphere,
preferrably nitrogen, and treating with n-butyl lithium/hexane, followed by the addition of (R)-
glycidyl butyrate and ~llowing the mixture to walm to 20-25. This method produces in high
yield the optically active 5-hydroxymethyl oxazolidinone having the desired stereochemistry (R),
i.e., the 5-,B configuralion, see EXAMPLE 14. `
This alcohol is then treated with triphenylphosphine and diethylazodicarboxylate in
tetrahydrofuran or ether, to give the optically active 5-phthalimidomethyl oxazolidinone, see
EXAMPLE 15.
EXA~LE 16 discloses that the 5-phthalimidomethyl compound is then treated with areagent to remove the phthalimide group such as an aqueous solution of methylamine or
hydr 7ine to give the intermediate 5-aminomethyl oxazolidinone. This is not purified, but is
immediately ,lcetylated by treatment with acetic anhydride or acetyl chloride in pyridine or
methylene chloride to give the (S)-5-acetamidomethyl oxazolidinone with the [carbo-t-butyloxy]- i
protecting group on ~ indolinyl nitrogen.
EXAMPLE 1~ discloses the removal of the protecting group by the addition of
trifluoroacetic ~cid, either neat, or in methylene chloride, to give the parent indoline
ox~zolidinone.
EXAMPLE 18 discloses acylation on the indoline nitrogen is effected by the addition of
~5 benzyloxyl~cetyl chloride in the presence of trieLhy!2~!ir.e or diisopropylethylamine in methylene
chk,ride hr ether, to give the benzyloxyacetylindolinyl compound. Trea~nent of this compound
2135~2 i , . ., ''
WO 94/01110 PCrlUS93/048~0
with hydrogen in the presence of palladium black or palladium/charcoal in ethyl acetate or
methanol (EXAMPLE 19) gives (S)-5-acetamidomethyl 3(5'-1-hydroxyacetylindolinyl)-
ox~zolidinone.
When the product of EXAMPLE 17 is acylated with 2-thiophenecarbonyl chloride
S (EXA~LE 20) the product is 5-(S)-N-(1'-(2-thiophenecarbonyl)-5'-indolinyl)-5- !-
(acetamidomethyl)-2-oxazolidinone .
The 5'-indolinyl oxazolidinones are administered either parenterally or orally. It is
known to those skilled in the art how to formulate ~e 5'-indolinyi oxazolidinones into
appropriate pharmaceutical dosage forms for oral (tablet, c~psule) or p~renteral (sterile solution)
use utilizing an appropriate solvent such as propylene glycol.
The 5'-indolinyl oxazolidinones are useful in treating tuberculosis in humans infected
with strains of M. tuberculosis which are susceptible to these antibiotics. The 5'-indolinyl
oxazolidinones are administered ~t doses from about 50 to about 3,000 mg/day. It is preferred
that the S'-indolinyl oxazolidinones a~ administered at doses of about 100 to about 2,000
mg/day. The 5'-indolinyl oxa~olidinones are administered orally and the tot~l d~ily dose is
divided between one to four doses per day.
The 5'-indolinyi oxazolidinones can either be used alone, or used in combination with
each other or with other antibiotics as is known to those skilled in the art. Preferred drugs tO be
used in combination with 5'-indolinyl oxazolidinones include, but are not limited to, isoniazid,
rifampin, ethambutol, p-aminosalicylic acid, pyra~inamide, streptomycin, kanamycin,
capreomycin, cyclosenne, and ethionamide. ',~
It is Icnown to those skilled in the art how to determine if humans are infected with M.
tubercl~losis, ~nd how to dete~nine if these organisms are susceptible to the 5'-indolinyl
ox~olidinones.
The exact dosage and frequency of administration depends on the particular 5`-indolinyl
ox~zolidinones used, the p~rticular condition being tre~ted, the severity of the condition being
tre~ed, the age, weight, general physical condition of the panticular patient, other medication the
individual may be taking as is well known to those skilled in the art and can be more accurately
de~ermined by measuring the blood level or concentration of the 5'-indolinyl oxazolidinones in
the patient's blood ~nd/or the patient's response to the palticular condition being treated.
DE~INITIONS AND CONVENTIONS ,~
The definitions and explanations below are for the terms as used throughout this entire
document including both the specification and the cl~ims.
DE~INlTlONS
All temperaNres are in degrees Centigrade.
~LC refers to thin-layer chromatograph
- - 213632~ `
WO 94/01110 PCr/US9'3/04850 ~
5 :,
THF refers to tetrahydrofuran.
Saline refers to an aqueous saturated sodium chloride solution.
When solvent pairs are used, the ratios of solvents used are volume/volume (v/v).
When the solubility of a solid in a s~lvent is used the ra~io of the solid to the solvent is
S weight/volume (wth).
IR refer~ to infrared spectroscopy.
[~]D25 refers to ~he angle of rota~ion of plant polarized light (specific optical rotation) at
25 with the sodium D line (5893A).
MS refers to mass spectrometry expressed as m/e or mass/charge unit. [M + H]~ refers
10 to the positive ion of a parent plus a hydrogen atom. El refers to electron irnpact. CI refers to
chemical ionization. FAB refers to fast atom bombardment.
Pharmaceutically acceptable refers to those properties and/or substances which are
acceptable to the patient from a pharmacologicalftoxicological point of view and to the
manufacturing pharmaceutical chemist fr~m ~ physical/chemical point of view regarding
15 composition, formulation, stabUity, patient acceptance and bioavailability.
M. tuberculosis refers to Mycobacteri~n tuberculosis.
EX~IP~ES
Without fur~er elaboration, it is believed that one skilled in the art can, using the
preceding description, practice the present invention to its fullest extent. The following detailed
20 examples describe how to prepare the various compounds and/or perform the various processes
of the invention and are to be constlued as merely iUustr~tive, and not limitations of the
preceding disclosure in ~ny way whatsoever. Those skilled in the arl will promptly recognize
appropriate variations frorn the procedures both as to reactants and ~s to re~ction conditions and
techniques.
25 EXA~LE 1 N-Benzyloxyace~yl-5-nitro-indoline (A)
Triethylamine (45 ml) is added to a solution of 5-nitroindoline (42.05 g) in
tetrahydrofuran (250 ml) and the mixture cooled to O, then a solution of benzyloxyacetyl
chloride (48.6 g) in THF (50 ml) is added over 15 min, and the mixn~re stirred in the ice bath
for an ~dditional 1.25 hr, then ~lowed to warm to 20-25 over 3 hr. Aqueous workup yields a
30 solid which is recrystallized fr~m acetone/w~er to give the title compound, mp 137-139.
EXAMPLE 2 N-Benzyloxyacetyl-5-amino-indoline (B)
To a mixture of ~-benzyloxy~cetyl-5-nitro-indoline (A, EXAMPLE 1, 12.26 g) in
methanol/ethyl acetate (18/82, 1.1 l) under nitrogen is added pa!ladium/charcoal (10%, 1.55 g)
and the fl~sk altemately evacuated and filled with hydrogen from a balloon (3 times), then the
35 mixture is allowed to stir at 2~25 for 4 hr. The mixture is degassed ~nd then filtered over
di~tom~eous ear~h ~nd the p~d washed with ethyl acet~te. The filtrate is concentra~ed to
21 3~ 3 2 ~
O 94/01110 ; ~ PC~/US93tO4850
-6-
volume of about 200 ml and a crystalline solid is precipi~ed, which is collected to give the title
compound, mp IOS-106.
EXAMPLE 3 N-Benzyloxyacetyl-5^~(2'-(~)-hydroxy-3'-butyrate)propylamino]indoline
(C)
S The following procedure is a modification of that described by M. alini, P. Crotti, and
F. Macchia, "Metal sal~ as new catalysts for mild and efficient aminolysis of oxiranes,"
Tetrahedron Letters, 31, 4661 (1990). The prior art process uses a full equiv~lent of zinc
chloride, but since i~ was found that the reaction stops due to complex formation, and side
re~ctions are more prevalent, 5 mole % of zinc chloride is used.
In a flame dned flask, zinc chloride (0.12 g) is dissolved in dry acetonitrile (10 ml),
then (R-)glycidyl butyrate (3.0 rnl) is added and the mixture stirred at 20-25, while N-
benzyloxyacetyl-S-amino-indoline (B, EXAMPLE 2, 4.94 g) in acetonitrile (65 ml) is slowly
added. The mixture is refluxed for a total of 2 days, then poured in~o 200 ml of water and
ex~acted with ethyl acetate (5 x 50 ml). The phases are separated and the organic layers dried
vith magnesium sulfate and concentrated under reduced pressure to give an oil residue. This is
carried on into the following reaction without purification; TLC analysis indicated a spot at Rf=
0.16 on silica gel, acetone/methylene chloride (10/90), for the desired compound. An analytical
sample is puzified using this system and gave a sample for MS, calcd for C2~H30O5N2 =
426.2155, found = 426.2153.
EXAlv~LE 4 5-(R)-N-(l'-benzyloxyacety1-5'-indolinyl)-5-(butyryloxymethyl)-2-
ox~zolidinone (D)
To a mixture of N-benzyloxyacetyl-5-[(2'-(R)-hydroxy-3'-butyra~e)propylamino]indoline
(C, EXAMPLE 3, 5.8 g) in methylene chloride (255 ml) under nitrogen at -15 is added
a solution of phosgene in toluene (1.93 M, 8.4 ml), and the mixture is allowed ~ slowly warm
to 0 over 1 hr. After an additiona~ lS min, dusopropylethylam~ne (5.7 ml) is added in
dropwise fashion over 5 min, and the mix~re stirred in the ice bath for 50 min, then allowed to
walm to 22, then poured into I N hydrochloric acid (SO ml). The layers are separated and the
aqueous is extracted with methylene chloride (3 x 25 ml). The combined organic layers are
washed successively with (75 ml ea) saturated aqueous sodium bicar~onate, water, and saline,
then dried magnesium sulfate, and concentrated under reduced pressure. The residue is purified
on silica gel (6 cm x 38 cm column, 40-63 y) with a gradient elution in acetone (2%-9%) in
methylene chloride. The appropriate fractions are pooled and concentrated to give the title
compound.
EXA~LE 5 S -(R)-N-( l ' -benzyloxyacetyl-S ' -indolinyl)-5 -~butyryloxymethyl)-2-
ox~zolidinone (D)
A mixture of N-benzyloxyacetyl-5-[(2'-(R)-hydroxy-3'-butyrate)propylarnino]indoline
:-- 2136324
WO 94/01110 PCI/US93fO4850
(C, EXAMPLE 3, ~ !j9 g) and carbonyldiimidazole (0.26 g) in THF (3 ml) is refluxed for 7
days, then an ~dditional carbonyldumidazole (0.26 g) and THF (0.5 ml) are added, and the
mixture refluxed for S hr. The mixture is poured into 0.5 N aqueous hydrochloric a~id, the
layers separated, and the aqueous extracted with ethyl acetate, and purification as above gives ;~
S the title compound, MS Calcd for C~5H2,~06N~ = 452.1947, found = 452.1940; IR (neat) 1742,
1668 cm '.
EXAMPLE 6 5-{R)-N-( I ' -benzyloxyacetyl-S ' -indolinyl)-5 -~ydroxymethyl)-2-
oxazolidinone ~E)
To a solution of S-(R)-N-(I'-benzyloxyacetyl-S'-indolinyl)-S-(butyryloxymethyl)-2-
10 oxazolidinone (D, EXAMPLE 5, 0.050 g) in methanol (I ml) is added sodium methoxide in
meth~nol (25%, 2.5 yl), and the mixture is stir~ed at 20 for I hr, then the mixture is `
concentrated to give a residue. Purification on a 250 lu silica gel plate in ethyl acetate (3
elutions) gives the title compound, TLC Rf = 0.28 (ethyl acetate); FAB MS Calcd for
C2lHnN2s = 382.1529, found = 382.1529.
EXA~LE 7 S~ N-(I'-benzyloxyacetyl-S'-indolinyl)-S-(azidomethyl)-2-
oxazolidinone (1~
Following the general procedure of US Palent 4,801,600 and making non-critical
vari~tions, 5 -(R)-N-( 1 '-benzyloxyacetyl-5 '-indolinyl)-S-(hydroxymethyl)-2-oxazolidinone (E,
EXAMPLE 6) is converted to the a~ide by treatment with methanesulfonyl chloride and
triethyl~nine in me~ylene ch~oride, followed by displacement with sodium
azide in refluxing acetone-water. This is immediately carried into the following reaction.
EXAMPLE 8 5-(S)-N-( 1 ' -benzyloxyacetyl-S ' -indolinyl)-S -(acetamidomethyl)-2- oxazolidinone (H)
Following the general procedure of EXAMPLE 2 and making non-critical va~i~tions but
t~king care to mon~tor the reduction of the azide so that unwanted cleavage of the benzyl group
does not take place, S-(R)-N-(1'-benzyloxyacetyl-S'-indolinyl)-S-(azidomethyl)-2-oxazolidinone
(F EXA~LF 7? is converted into an amine (G) by treatment witil palladiwn/charcoal in 1 atm
of hydrogen in ethyl acetale. Removal of the hydrogen gas and purging with nit~gen is
followed by the ~ddition of pyridine and acetic anhydride (2 mole equiv. each), and the reaction
stirred at 20-25 for 12 hr, then filtered over dia~omaceous earth, and the filtrate concentrated.
The residue is purified by column chromatography using a gradient of methanol/ethyl acetate to
give the title compound.
EXAMPLE 9 5-(S)-N-( I '-Hydroxyacetyl-5 ' -indolinyl)-5-(acetamidomethyl)-2-
ox~zolidinone (I)
5-(S)-N-( I '-benzyloxyacetyl-5 ' -indolinyl)-5-(acet~rnidomethyl)-2-oxazolidinone (H,
EXAhJfPLE 8) is dissolved in ethyl acetate and the mixture purged with nitrogen gas, then
213632~ ~
WO 94/0111~ ' PC~/lJS93/0485
-8-
pal!adium/charcoal is added followed by hydrogen (balloon) and the mLxture is stirred until TLC
analysis shows eomplete conversion of the st~ng material. The mixture is filtered over
di~tom~eous earth, concen~ated and the residue is purified by colwnn chromatography on
silic~l gel (methanol/ethyl acetate) to give the title compound.
5 EXA~LE 10 N-(t-Butyloxycalbonyl)-5-[(2'-(R)-hydroxy-3'-butyra~e)propylamino]-
indoline (J3
Following the general procedure of EXAMPLE~ 3 and making non-critical variations but
starting with N-(t-butyloxycarl~onyl3-5-aminoindoline~ the ~tle compound is obtained.
EXA~LE 11 5-(S)-N-(l'-(t-Bucyloxycar~onyl)-5'-indolinyl)-5-(acetamidomethyl)-2-
10oxazoUdinone (K)
Following ~e general procedure of EXA~LES ~8 and making non-cri~ical variations
- but st~ing with N-(t-butyloxycarbonyl)-5-~(2'^(R)-hydroxy-3'-butyrate)pTopylamino]indoline
(EXA2~LE 10), the title compound is obtained.
EXA~LE 12 5(S)-N-(5'-in~olinyl)-5-(acetamidomethyl~-2-oxa~olidinone (L)
15A 2~fold excess of trifluoroacetic acid is added slowly over a period of about 5 min to
5-(S)-N-( I '-(t-butyloxycarbonyl)-5 '-indolinyl)-5-(acetamidomethyl)-2-oxazolidinone (K,
EXA~LE 11) in methylene chloride at 0. The mixture is stirred for 3 hr at th~t temperature.
Then the mixture is neutralized by the slow ~ddition of s~ed aqueous sodium bicarbonate,
the layers separated, and the aqeous phase is extracted with methylene chloride to give the title
20 compound.
EXAMPLE 13 5-(S)-N-(1'-(2-Thiophenecarbonyl~-5'-indolinyl)-5-(acetamidomethyl)-2- ~-
oxa~olidinone (P)
Following the general procedure of Intemational Publica~on No. W090/~2744,
EX~LE 18, and making non-critic:ll variations and s~ing with 5(S)-N-(5'-indolinyl)-5-
25 (acetamidomethyl~-2~xazolidinone (L, EXA~LE 12) and using the ~ppropriate corresponding
acyl~ting agent the title compound is obtained.
EXA~I,E 14 (R)-3-(5'-1-Carbo-t-butyloxyindolinyl)-5-(hydroxymethyl)-oxazolidin-2-
one
n-Butyl lithium/hexane (1.6 M, 16.7 ml) is added dropwise over 5 min to a mixture of
30 I-carbo-t-butyloxy-tN-carboberlzyloxy)-S-aminoindoline (IV, US Patent 5,164,510, EXAMPLE
11, 9.371 g) in freshly distilled tet~hydrofu~n (140 ml) at -78 under nitrogen. The mixture is
stirred for 10 min, then ~R)-glycidyl butyrate (4.0 ml) is added via syringe, the mixture is stirred
for 1 hr at -78, then slowly ~llowed to warm to 20 overnight.
S~ur~ted aqueous ammonium chloride (100 ml) and water (200 ml) is added to the mixture and
35 this mixture is extracted with ethyl acetate (125 ml, then 3 x 100 ml), and the combined orgaruc
laye~ are washed wiuh saline, and dried over magnesium sulfate. The mixture is filtere asld the
--- 213S~
.. . .
WO 94/01110 PCrlUSg3/04X50
filtr~te is concentraled to give a solid which is recrystallized from hot ethyl acetate/hexanes to
give the title compound, mp 15~157; []D=40 (CHClt).
EXAMPLE 15 ~R)-3-(St-l-Carbo-t-butyloxyindolinyl)-5-(phthalimidomethyl)-oxa~olidin-
2~ne
Diethylazodicarboxyla~e (3.45 mL) is added over 3 min to a mixture of (R)-3-(5'-1-
c~rbo-t-butyloxyindolinyl)-5-(hydroxymethyl)-oxæolidin-2-one (EXA~LE 14, 7.103 g),
phthalimide (3.228 g) and triphenylphosphine (5.804 g) in freshly distilIed tetrahydrofuran (175
ml) ~t Oo under nitrogen. The mixture is s~irred at 0 for 45 min, then allowed to come to 20
overnight, and the volatile components removed on a rotary e~porator. The residue is purified
by cluomatography on silic~ gel 4~63 microns, 4.5 cm (height) x 10.0 cm (diameter) column,
eluting with methylene chloride. The appropriate fractions are pooled and concentJated to give
- ~e title compound, [alD = -52 (CH3CN); MS (EI, rela~ve abumdance) 463 (M~, 7.5).
EXAMPLE 16 (S)-3-(5'-1-Carbo-t-butyloxyindolinyl)-S-(acetamidomethyl)-oxazolidin-2-
one (K)
A mixture of (R)-3-(5'-1-carbo-t-butyloxyindolinyl)-5-(phthalimidomethyl)-oxazolidin-2-
one (EXA~LE 15, 8.826 g) in absolute ethanol (100 mi) and aqueous methylamine (40%, 50
ml) is heated to reflux under nitrogen for E5 hr. The vola~les are then removed by ~educed
pressure with heat at 0.1 Torr to give a concentrate. To this concentrate is ~dded 50 ml of
pyndine and 25 ml of acetic anhydride at 20. A slight exotherm occurred, so the flask is
cooled in a water bath to maintain the temperature at about 30. After stining for 1.3 hr, the
volatiles were removed by vacuum distillation (0.1 Torr), giving a resîdue. This residue is
chrom~togr~phed on silica gel 40 63 micron, 9.6 cm dia x 4 cm height, eluting with a gradient
of ~cetone in methylene chloride (0-50%, v/v). The appropriat fractions are pooled and
concentrated to give a the title compound, mp = 153.5-155~
EXAMPLE 17 (S)-3-~5'-indolinyl)-5-(acetamidomethyl)-oxa~oUdin-2-one (L)
Trifluoroacetic acid (5 ml) is added to a mixture of (S)-3-(5'-l~arto-t-
butyloxyindoUnyl)-S-(acetamidomethyl)-oxa~olidin-2-one (K, EXAMPLE 16, 4.705 g)
in methylene chloride (40 ml) at 0 under nitrogen ~nd the ice bath removed. After 4 hour, the
mix~re is concentrated and methylene chloride (8 ml) and trifluo~acetic acid (7 ml) is added at
20-25, After stin~ng for an additional 2 hr, the mixture is concentrated under reduced pressure,
and ~he residue is poured into saturated aqueous sodiurn bicarbonate and ice, and the mixture is
continuously extracted with methylene chloride (500 ml) for 24 hr. The organic phases are
combined, dried over ma~esium sulfate and concentrated to give the title compound, mp 47-
50; TLC R~ = 0.11, methanol/ethyl acetate (10/90).
EXAMPLE 18 (S)-3-(5'-l-Benzyloxyacetylindolinyl)-5-(acetamidomethyl)-oxa~olidin-2-
one
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WO 94/01110 PCr~US93/04850
-10-
Benzyloxyacetyl chloride (2.27 rnl) is added dropw~se over 5 m~n to a mixture of (S)-3-
(5'-indolinyl)-5-(acetamidomethyl)-oxazolidin-2-one (L~ EXAMPLE 17, 3.445 g) andtriethylarnine (2.62 ml) in methylene chloride (100 ml) at 0 under r~itrogen. The mixture is
slowly allowed to come to 20 overnight. The mixture is cooled to 0, and filtered. the collected
5 sn~terial is washed with water (2 x 50 rnl) and dried in a vacuum oven at 70. The solid is
recryst~llized frotn hot methylene chloride and hex~nes to give the title compound, mp 188-
189, [a]D = -13 (CHCI3).
EXAMPLE l9 (S)-3-(5'-1-Hydroxyacetylindolinyl)-5-(acetamidomethyl)-oxazolidin-2-
one (I)
A reaction flask holding a mixture of (S)-3-(5'-1-benzyloxyacetylindolinyl)-5-
(acetamidomethyl)-oxazolidin-2-one (~XAMPLE 18, 9.119 g) in methanol/methylene chloride
(10/90, 500 ml) is evacuated and filled with nitrogen gas three times, then 0.96 g of palladium
bl~ck is added, and the flask again evacuated and filled with nitrogen three times, then
ev~cuated and filled with hydrogen from a balloon (three t mes). Af~er stirring of ~e mixture ~
20-25 for a total of 4 hr, the hydrogen is removed and the mixture is filtered over diatomaceous
ea~ nd the filtlate was concentrated in vacuo to give a solid. Illis solid is ~iturated with
methanol/ethyl acetate (10/90, 200 ml) in a 35 bath, then cooled ~ 20C and the solid collected
to give the title compound, mp 198-199, la~D = -210 (DMSO).
EXAMPLE 20 (S)-3-(5'-1-((2-Thiophenecarbonyl)indolinyl)-5-(acetamidomethyl)-
oxazolidin-2-one (P)
Triethylamine (2.5 ml) followed by 2-thiophenecarbonyl chloride (1.5 ml) is added over
3 min to a mixture of (S)-3-(5'-indolinyl)-S-(acetamidomethyl)-oxazolidin-2-one (L~ EXAMPLE
17, 3.289 g) in me~ylene chloride (70 ml) under nitrogen at 0. The mixture is allowed to
come to 2~25 overnight. The mixture is filtered, and the solid is washed with methylene
chloride, followed by water, and then dried in a vacuum oven at S0. The solid is recr,vstallized
from acetone-water to give the title compound, mp = 193-198, [a]D = -38
(dimethylformamide).
EXAMPLE A 58 Year Old, 80 kg Male lnfected With M. tuberculosis
A 58 yr old, 80 kg white m~le seeks medical attention due to an illness characterized by
cough, weakness, night sweats ~nd shortness of breath The patient's chest ra~iograph shows
extensivè cavitary disease consistent wi~ a di~gnosis of tuberculosis. Spu~m cultures are
positive for M. tuberculosis, and the organism is found to be resistant to isonia~id, rifampin, and
streptomycin. The patient is admitted to the hospital and given 5(S)-N~ hydroxyacetyl-5'-
indolinyl)-5-(acetamidomethyl)-2-oxazolidinone orally at a total dosage of 2000 mglday for 4
weeks. Signs and symptoms of tuberculosis slowly disappear, viable M. tubercu~osis organisms
are no longer isolaled from the patients sputum, and the patients chest radiograph retums to
213~32~ ",
WO 94/01 1 10 ~ PCr/US93lO4850
norm~l. The patient is discharged f;rom the hospit~l.
EXAMPLE B 30 Year C)ld, 60 Kg Female Infeeted With M. ~uberc-~Jos~s
A 30 yr old, 60 kg black female is diagnosed with pulmonary tubera~losis. Her sput
isolate is detennined to be susceptible to ~oth isonia~id and p-aminosalicylic acid. She is
5 ~reated as an outpatient with a regimen of oral isonia2id and p-aminosalicylic acid, however,
retums in 3 weeks because her condition does not improve. A new sputum culture grows M.
tuberculosis that is resistant to isoniazid. The pa~ent is given a S(S)-N-(I'-hydroxyacetyl-5'-
indolinyl)-5-(acetamidomethyl)-2-ox~olidinone to take orally a~ a dose of lQOO mg/day for two
months, in addition to the other antibiotics. Within one month the signs and symptoms of
10 tuberculosis slowly disappear. Sputum cultures taken 3, 6 and 12 months post-therapy are
negative for M. ~uberclllosis.
EXAMPL~ C 36 Year Old Male With AIDS Infect~d With M. ~uberculosis
A 36 yr old white male with acquired immune deficiency syndrome (AIDS) presents to
his physician with fever, weight loss, and cough. The chest ~adiograph reveals a focal nidus of
15 infection in the lung. Expectorated sputum is negative for M. tuberculosis, however, a tissue
biopsy taken via bronchoscopy contains culturable M. tuberculosis ln spite of a susceptibility
report indicating the isolate is susceptible to the common therapy of isoniazid and rifampin, the
patient is placed upon th~e drug therapy (isoniazid, rifampin, and ethambutol) due to his AIDS
condition. The patient initially improves, but then his condition begins to ~lapse. A new
20 spu~rn culhlre indicates the presence of M. ~u~erculosis that has developed ~sistance to
rifampin, but is susceptible to 5(S)-N-(1'-hydroxyacety1-5'-indolinyl)-5-(acetamidomethyl)-2^
ox~zolidinone. The rifampin portion of the regimen is replaced by 5-(S)-N-(l'-hydroxyacetyl-5'-
indolinyl)-5-(acet~nidomethyl)-2-ox~zolidinone given at an oral dose of lOOO mg/day. Within 3
weeks the signs and symptoms of tuberculosis are resolved; the patient continues to take the
25 three drug regimen for an additional 8 months. A sputum culture taken post-therapy is negative
for M. tuberculosis The patient is then given isoniazid p~phylactically for life to prevent
recurrence of the infectio~
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