Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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BICYCLIC HYDANTOIN DERIVATIVES
AND COMBINATORIAL LIBRARIES THEREOF
BACKGROUND OF THE INVENTION
FIELD OF THE INVENTION
The present invention relates generally to the
synthesis of compounds comprising heterocyclic rings. In
one embodiment, the invention provides novel bicyclic
hydantoin derivative compounds as well as novel
combinatorial libraries comprised of such compounds.
BACKGROUND INFORMATION
The process of discovering new therapeutically
active compounds for a given indication involves the
screening of all compounds from available compound
collections. From the compounds tested, one or more
structures is selected as a promising lead. A large
number of related analogs are then synthesized in order
to develop a structure-activity relationship and select
one or more optimal compounds. With traditional "one-at-
a-time" synthesis and biological testing of analogs, this
optimization process is long and labor intensive. Adding
significant numbers of new structures to the compound
collections used in the initial screening step of the
discovery and optimization process cannot be accomplished
with traditional "one-at-a-time" synthesis methods,
except over a time frame of years or even decades.
Faster methods are needed that allow for the preparation
of up to thousands of related compounds in a matter of
days or a few weeks. This need is particularly evident
when it comes to synthesizing more complex compounds,
such as bicyclic hydantoin derivative compounds.
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Combinatorial approaches have been extended to
"organic," or non-peptide, libraries. Combinatorial
chemical methods have even been extended to hydantoin
derivative compounds, as described, for example, in
DeWitt et al., Proc. Natl. Acad. Sci. USA, 90:6909-6913
(1993); Dressman et al., Tetr. Lett., 37:937-940 (1996);
Hanessian et al., Tetr. Lett., 33:5835-5838 (1996); Kim
et al., Tetr. Lett., 38:4603-4606 (1997); Yoon et al.,
Chem. Commun., 2703-2704 (1998); Wilson et al., Tetr.
Lett., 39:5135-5138 (1998); Chong et al., Tetr. Lett.,
40:2493-2496 (1999); Wu et al., Tetr. Lett., 41:1165-1169
(2000); Park et al., J. Org. Chem., 63:113-117 (1998);
Gong et al., Tetr. Lett., 39:3379-3382 (1998). Gong et
al., J. Org. Chem., 63:3081-3086 (1998); and Gong et al.,
J. Org. Chem., 63:4854-4856 (1998).
However, the libraries to date contain compounds
of limited diversity and complexity. Such compounds are
particlularly limited regarding amino and substituted
amino radicals attached to the bicyclic hydantoin core.
A reed therefore exists to develop more complex
libraries based on heterocyclic medicinal compounds which
would need less time and effort in the synthesis and
testing required to bring an organic pharmaceutical
product to fruition. In short, improved methods for
generating therapeutically useful heterocyclic compounds,
such as bicyclic hydantoin derivatives, are desired.
Hydantoin derivative compounds have been the
subject of investigation in a number of different
biological areas. For example, hydantoin derivatives
have been proposed or used as anticonvulsant,
antibacterial, antidiabetic, antiarrhythmic, antifungal
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or herbicidal agents. See Spinks et al., Prog. Med.
Chem., 3:313 (1963); Wright et al., J. Med. Chem., 12:
379-381 (1969); Carrera et al., J. Heterocyclic Chem.,
29:847-852 (1992); Coudert et al., Pharm. Acta. Helv.,
66:155-159 (1991); Karolakwojciechowska et al.,
Pharmazie, 50:114 (1995) ; Issartel et al., Eur. J. Med.
Chem., 31:717-723 (1996); Nam et al, Arch. Pharm., 330:
268-270 (1997); U.S. Pat. No. 4,198,423, 1980 (BASF A.-
G., Fed. Rep. Ger.); and Mappes et al, Chem. Abstr., 93:
71784 (1980) .
Bicyclic hydantoin derivatives have been the
subject of serial chemical synthesis. See, for example,
WO 98/2721; Nam et al., supra.; WO 96/16111; Issartel et
al., supra.; WO 93/2413; Evans et al., J. Med. Chem., 36:
3993-4005 (1993); EP 91-810980; and JP 78-114787.
However, more complex bicyclic hydantoin derivatives,
especially those with a nitrogen substitution (i.e.,
amino, (mononsubstituted)amino or (disubstituted)amino)
on the non-hydantoin ring, have been difficult to attain
even through serial methods.
This invention satisfies this need and provides
related advantages as well. The present invention
overcomes the known limitations to classical serial
organic synthesis of bicyclic hydantoin derivatives, for
example, as well as the shortcomings of combinatorial
chemistry related to bicyclic hydantoin derivatives. The
present invention allows for rapid generation of large
diverse libraries of complex bicyclic hydantoin
derivatives as discrete molecules. The present invention
can utilize a readily available pool of building blocks
that can be incorporated into the various regions of the
molecule. Furthermore, the method of making the present
invention allows for the use of building blocks that
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contain a wide range of diverse functionality. Such
building blocks can provide combinatorial libraries that
consist of large numbers as well as combinatorial
libraries that are extremely diverse with respect to the
functionality contained within those libraries. The
present invention combines the techniques of solid-phase
synthesis of bicyclic hydantoin derivatives and the
general techniques of synthesis of combinatorial
libraries to prepare highly diverse new bicyclic
hydantoin derivative compounds.
SUMMARY OF THE INVENTION
The present invention relates to novel bicyclic
hydantoin derivative compounds of the following formula:
R
R
N R5
R
wherein R1 to RS and n have the meanings provided herein.
The invention further relates to combinatorial
libraries containing two or more such compounds, as well
as methods of preparing bicyclic hydantoin derivative
compounds.
R~
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BRIEF DESCRIPTION OF THE DRAWING
Figure 1 shows Scheme 1 for the combinatorial
synthesis of bicyclic hydantoin derivative compounds,
where n is 0 and the depicted exocyclic nitrogen is
5 attached to a sulfonyl derivative (-S (0,) R?) . The
reagents and conditions for each step of Scheme 1 are as
follows : step ( a ) : t-Boc-NH-Ri-CO~H, DIEA
(N,N-diisopropylethylamine), DIC
(diisopropylcarbodiimide), HOBt (1-hydroxybenzotriazole)
and DMF (N,N-dimethylforamide) for 20 hours; step (b) 550
TFA (trifluoroacetic acid) and DCM (dichloromethane) for
30 minutes; step (c) R,SO,C1, DIEA and DCM for 48 hours;
step (d) trans-L-t-Boc-4-Hyp-OMe, (hyp = hydroxyp.roline),
Ph~P (triphenylphosphine), DIAD, (diisopropyl
azodicarboxylate), NMM (N-methylmorpholine), DCM and THF
(tetrahydrofuran) for three days; step (e) 55% TFA/DCM
for 30 minutes; step (f) R~NCO and DMF for 3 days; step
(g) 0.025 M TMG/DMF (tetramethylguanidine) for 16 hours;
and step (h) gaseous HF (hydrofluoride) for 2 hours.
Figure 2 shows Scheme 2 for the combinatorial
synthesis of bicyclic hydantoin derivative compounds,
where n is 0 or l, Rz or R~ is -OH (as shown between steps
c and d) and the depicted exocyclic nitrogen is attached
to a sulfonyl derivative (-S(O2)R11; as shown between
steps d and a and thereafter). The reagents and
conditions for each step of Scheme 2 are as follows: step
(a) : t-Boc-NH-R1-COZH, DIEA, DIC, HOBt and DMF for 20
hours; step (b) 55o TFA/DCM for 30 minutes; step (c)
RaSO,Cl, DIEA and DCM for 48 hours; step (d) for three
days, Ph3P, DIAD, NMM, DCM, THF and ( i ) for n=0, a
4-hydroxy-N-Boc-proline ester derivative; (ii) for n=1, a
4- or 5-hydroxy-N-Boc-pipecolinate derivative; step (e)
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55 o TFA/DCM for 30 minutes; step ( f ) R~,NCO and DMF for 3
days; step (g) 0.025 M TMG/DMF for 16 hours; and step (h)
gaseous HF (hydrofluoride) for 2 hours.
Figure 3 shows Scheme 3 for the combinatorial
synthesis of bicyclic hydantoin derivative compounds,
wherein n is 0, 1 or 2 and using an alternate scheme,
resin and resin attachment position. The reagents and
conditions for each step of Scheme 3 are as follows: step
(a): DIEA, DIC (N,N'-diisopropylcar.bodiimide) HOBt and
DMF for 20 hours; step (b) 20o piperidine in DMF or 550
TFA/DCM for 30 minutes; step (c) RSNCO, DIEA and DMF for
hours; step (d) catalytic TMG/DMF or catalytic barium
dihydroxide in DMF.
DETAILED DESCRIPTION OF THE INVENTION
15 The present invention provides compounds and
combinatorial libraries of compounds of the formula:
R
R
N R5
Ra
R~
wherein:
n is 0, 1 or 2;
a) R1; b) when n is 0, R~; c) when n is 1, R4 and one of Re
20 and R3; and d) when n is 2 an addition radical between R3
and R4, R34, is present and, when n is 2, R4 and two of Rz,
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R~ and R~4:
are each independently a hydrogen atom, C1 to C1z alkyl, C_
to C1~ alkenyl, C7 to Clz alkynyl, C1 to Ci, substituted
alkyl, C~ to C1; substituted alkenyl, C~ to Ci~ substituted
alkynyl, C~ to C~ cycloalkyl, C~ to C.; substituted
cycloalkyl, CS to C~ cycloalkenyl, CS to C~ substituted
cycloalkenyl, heteroaryl, substituted heteroaryl, C, to
Clb phenylalkyl, C~ to C18 substituted phenylalkyl, phenyl,
substituted phenyl, naphthyl, substituted naphthyl,
cyclic C~ to C., alkylene, substituted cyclic C, to C~
alkylene, cyclic C~ to C, heteroalkylene, substituted
cyclic C~ to C7 heteroalkylene, amino,
(monosubstituted)amino, (disubstituted)amino, protected
(monosubstituted)amino, carboxy or protected carboxy; and
a) when n is 0, R,; b) when n is 1, one of Rz and R3; and
c) when n is 2 one of R~, R~ and R~4:
is amino, (monosubstituted)amino, (disubstituted)amino,
protected (monosubstituted)amino or the formula:
-N-R-,-R~
Rs
wherein:
R6 is the formula -L-M, wherein -L- is -C(0)-, -C(O)O-
or -S (0) ~- and M is a hydrogen atom, C1 to C1, alkyl, C1
to C12 substituted alkyl, C~ to C1z alkenyl, C~ to C12
substituted alkenyl, phenyl, substituted phenyl,
naphthyl, substituted naphthyl, C,.to Cla phenylalkyl,
C, to C18 substituted phenylalkyl, C, to C1~
heterocyclicalkyl, C; to C1~ substituted
heterocyclicalkyl, heteroaryl, substituted heteroaryl,
heterocycle or substituted heterocycle;
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R; is the formula -D-W-E-, wherein at least one of D, W
and E is present and the other two are optionally
present or absent, and wherein:
W is C~ to C.; cycloalkylene, C; to C, substituted
cycloalkylene, C~ to C~ cycloalkenylene, C~. to C;
substituted cycloalkenylene, arylene, substituted
arylene, heterocyclene, substituted heterocyclene,
heteroarylene or substituted heteroarylene; and D,
which, if present, is directly attached to the nitrogen
depicted in the formula, and E are independently C1 to
C1, alkylene, C~ to C1, alkenylene, C~ to C" alkynylene,
C1 to C12 substituted alkylene, C~ to C,_ substituted
alkenylene or Ca to C1, substituted alkynylene;
R~ is a hydrogen atom, a halide, -OR9, -CO;R;, -C (O)-NRgRio
or -NR9Rlo, wherein R9 and Rlo are independently a
functionalized resin, a hydrogen atom, C, to C1, alkyl,
C1 to C1~ substituted alkyl, phenyl, substituted phenyl,
heterocycle, substituted heterocycle, heteroaryl,
substituted heteroaryl, C3 to C; cycloalkyl, C3 to C,
substituted cycloalkyl, CS to C, cycloalkenyl, CS to C-,
substituted cycloalkenyl, C, to C18 phenylalkyl, C; to
C1~ substituted phenylalkyl, C1 to C12 heterocyclicalkyl,
C1 to Clz substituted heterocyclicalkyl, C1 to C12 aryl,
C1 to C12 substituted acyl, phenylsulfonyl, substituted
phenylsulfonyl, C1 to Clo alkylsulfonyl, C1 to Clo
substituted alkylsulfonyl, C1 to C1~ alkylaminocarbonyl,
C1 to C22 substituted alkylaminocarbonyl,
phenylaminocarbonyl, substituted phenylaminocarbonyl,
C1 to Clz alkylaminothiocarboriyl, C, to C1, substituted
alkylaminothiocarbonyl, phenylaminothiocarbonyl or
substituted phenylaminothiocarbonyl; and
RS is a hydrogen atom, C1 to C1z alkyl, C1 to C,
substituted alkyl, C, to C1, alkenyl, Ca to C,, substituted
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alkenyl, Ca to C1, alkynyl, C, to C1_, substituted alkynyl,
phenyl, substituted phenyl, naphthyl, substituted
naphthyl, C~ to Clu phenylalkyl, C, to C1~ substituted
phenylalkyl, C~ to C, cycloalkyl, C~ to C, substituted
cycloalkyl, CS to C, cycloalkenyl, CS to C~ substituted
eycloalkenyl, C1 to C1, heterocycloalkyl, C1 to Clz
substituted heterocycloalkyl, C1 to Cl~ alkylsulfonyl, C1
to Clo substituted alkylsulfonyl, C, to C1,
alkylaminocarbonyl, Cl to C12 substituted
alkylaminocarbonyl, heterocyclic ring or substituted
heterocyclic ring, or (i) the formula -C(O)R11; (ii) the
formula -C (O) OR11; (iii) the formula -C (O) NHR11; (iv) the
formula -C (O) NR11R1~; or (v). the formula -S (O~) R11, wherein
R11 and R1, are, independently, a hydrogen atom, C1 to Clz
alkyl, C, to C1, substituted alkyl, C~ to C1, alkenyl, C2 to
Cla substituted alkenyl, C, to C1? alkynyl, C: to Cla
substituted alkynyl, phenyl, substituted phenyl,
naphthyl, substituted naphthyl, C, to C1~ phenylalkyl, C~
to C1~ substituted phenylalkyl, C3 to C~ cycloalkyl, C3 to
C., substituted cycloalkyl, C, to C~ cycloalkenyl, C5 to C-,
substituted cycloalkenyl, C1 to C1~ heterocycloalkyl, C1 to
C1~ substituted heterocycloalkyl, heterocyclic ring or
substituted heterocyclic ring, or R5, together with the
adjoining nitrogen atom depicted in the formula, form a
heterocyclic ring or substituted heterocyclic ring,
wherein said ring is non-aromatic.
In a further embodiment of the present invention,
n is 0.
In an additional embodiment, R1 and R3 are each a
hydrogen atom.
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In another embodiment,
R, is the formula:
-N-R-,-Re
5 R6
wherein:
R6 is the formula -L-M, wherein -L- is -S(O)z- and M is
phenyl, substituted phenyl, naphthyl, substituted
naphthyl, heteroaryl or substituted heteroaryl;
10 R7 is the formula -D-W-, wherein D is present and W is
optionally present or absent, and wherein:
D is C, to C1, alkylene or C1 to C:_ substituted alkylene;
and W is C3 to C~ cycloalkylene, C; to C, substituted
cycloalkylene, arylene or substituted arylene; and
RF is -C (O) NR9Rlo, wherein one of R5 and R,~, is a
functionalized resin and a hydrogen atom, and the other
is a hydrogen atom.
In an additional embodiment,
RS is C1 to C12 alkyl, C1 to C1~ substituted alkyl, phenyl,
substituted phenyl, naphthyl, substituted naphthyl, C, to
Cle phenylalkyl or C, to C18 substituted phenylalkyl.
A further embodiment of the present invention
provides a combinatorial library or single compound
wherein:
n is 0;
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R1 and R; are each a hydrogen atom;
R; is the formula:
-IV-R,-R8
I
R6
wherein:
R6 is the formula -L-M, wherein -L- is -S(0)z- and M is
thiophen-2-yl, phenyl, 2,5-dichlorophenyl, 2-
nitrophenyl, 4-bromophenyl, 4-fluorophenyl, 4-
chlorophenyl, 3-(trifluoromethyl)phenyl, 3,4-
dichlorophenyl, 3-chloro-4-fluorophenyl, 2-
fluorophenyl, 3-fluorophenyl, 4-
(trifluoromethyl)phenyl, 2,4-difluorophenyl, 2-
chlorophenyl, 2-(trifluoromethyl)phenyl, 3=
chlorophenyl, 3,5-dichlorophenyl, 2,3-dichlorophenyl,
2-bromophenyl, 5-(2-pyridyl)thiophen-2-yl, 2-chloro-5-
(trifluoromethyl)phenyl, 4-cyanophenyl, 2-cyanophenyl,
5-chloro-1,3-dimethylpyrazol-4-yl, 3,5-
dimethylisoxazol-4-yl, 2,4-dichlorophenyl, 2-chloro-4-
(trifluoromethy)lphenyl, 2-chloro-4-fluorophenyl,
2,4,6-trichlorophenyl, 1-methylimidazol-4-yl, 2-
methoxycarbonythiophen-3-yl, 5-(isoxazol-3-yl)thiophen-
2-yl, 4-phenylphenyl, 3,4-difluorophenyl, 3-methyl-5-
chlorobenzothiophen-2-yl, 3-cyanophenyl, 4-
methylsulfonylphenyl or
2-methylsulfonylphenyl;
R7 is the formula -D-W-, wherein:
W is absent and D is phenylethyl-1-ene, ethyl-1-ene,
propyl-1-ene, propylene, pentylene, 4-
(chlorophenyl)ethyl-1-ene, 3-(methylthio)propyl-1-ene,
4-(methoxyphenyl)ethyl-1-ene, 2-methylpropyl-1-ene, 2-
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(4-imidazole)ethyl-I-ene, 2-(benzyloxy)ethyl-1-ene, 3-
methylbutyl-1-ene or 2-cyclohexylethyl-1-ene; or D is
methylene and W is 4-phenylene or 4-cyclohexylene; and
R8 is -C (O) NHS; and
RS is phenyl, 2-bromophenyl, 2-fluorophenyl, 2,4-
difluorophenyl, 2,6-difluorophenyl, 2-chlorophenyl, 2,4-
dichlorophenyl, 2,5-dichlorophenyl, 2-methoxyphenyl, 0-
tolyl, 2-ethylphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-
methoxyphenyl, m-tolyl, 4-bromophenyl, 4-fluorophenyl, 4-
methoxyphenyl, p-tolyl, 1-naphthyl, benzyl, 2-
isopropylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl,
2-ethyl-6-methylphenyl, 3-(methylthio)phenyl, 3,4-
dimethylphenyl, 3,5-dimethylphenyl, 2-methoxy-5-
methylphenyl, 3-ethylphenyl, 4-ethoxyphenyl, 4-
(methylthio)phenyl, 4-isopropylphenyl, 4-ethylphenyl, 4-
n-butylphenyl, 2-isopropyl-6-methylphenyl, 2,4,5-
trimethylphenyl, 4-butoxyphenyl, 5-fluoro-2-methylphenyl
or 4-(dimethylamino)phenyl.
The invention also provides methods of
preparing bicyclic hydantoin derivative compounds and
combinatorial libraries. In one method, as shown in
Figure l, such compounds can be prepared by (1) coupling
(a) a molecule containing a group of the formula -NH-
C (0) -RI-NH-S (Oz) -Rz, wherein Rz and R2, independently, are
each a variable group, with (b) a ring nitrogen compound
that is substituted at the position adjacent to the ring
nitrogen (the 2 position) with -C(O)-O-alkyl, where the
alkyl group is preferably ethyl or methyl, and where the
ring is further substituted with a hydroxy group,
resulting a ring nitrogen compound that is substituted at
the position adjacent to the ring nitrogen with
-C(O)-O-alkyl and further substituted with a group of the
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formula -N (S (0=) -R;) -R1-C (0) -NH- (see step d of Figure 1
and step d of Figure 2); (2) reacting the resulting
compound of step (1) with an isocyanate of the formula
Rz-NCO, where R= is a variable group, to form a
-C(0)-NH-R~ group attached to the ring nitrogen
(1 position); and (3) cyclizing the resulting compound of
step (2) by reacting it with a base to form a bicyclic
hydantoin derivative. Exemplary bases include
tetramethylguanidine and barium hydroxide.
The method further provides where the molecule
containing a group of the formula -NH-C(O)-R1-NH-S(Oa)-R=
is attached to a functionalized resin. The method also
provides that the molecule containing a group of the
formula -NH-C (O) -R1-NH-S (O,) -R~ is formed by coupling a
molecule containing a group of the formula -NH-C(O)-R1-NH,
with R.,-S(O,)-leaving group, preferably, where the leaving
group is a halide and, more preferably, where the halide
is chloride.
The method additionally provides where the ring
nitrogen compound is a pyrrolidine derivative and,
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therefore, the resulting compound of step (3) is of the
formula:
n
O
N N-Rs
H N-
O
The method also provides where the ring
nitrogen compound is a piperidine derivative (e. g, a six-
member ring) and where it is a seven-member ring. It
should be understood that the ring nitrogen can be fused
with another ring, for example, an aromatic ring. It
should also be understood that, where, the ring is six or
seven member, it can contain a double bond. However, the
ring should not be aromatic.
Another method of the present invention
provides (1) coupling (i) a ring nitrogen compound,
wherein said ring is non-aromatic, wherein said ring is
attached to a group containing -C(0)-0-, said ring
directly attached from a position adjacent to the ring
nitrogen to the carbonyl carbon of said -C(0)-0-, with
(ii) an isocyanate derivative of the formula variable-NCO
to form (iii) a ring nitrogen compound with the group -
C(0)-NH-variable directly attached to the ring nitrogen;
and (b) cyclizing the resulting compound of step (1) by
reacting it with a base to form a bicyclic hydantoin
derivative. Exemplary bases include tetramethylguanidine
and barium hydroxide.
The method further provides where the group
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containing -C(O)-O- is attached to a functionalized resin
(via oxygen). The method also provides where the ring
nitrogen is five- six- or seven-member (see Figure 3).
It should be understood that the ring nitrogen can be
5 fused with another ring, for example, an aromatic ring.
It should also be understood that, where, the ring is six
or seven member, it can contain a double bond. However,
the ring should not be aromatic.
When the above-described compounds include one
10 or more chiral centers, the stereochemistry of such
chiral centers can independently be in the R or S
configuration, or a mixture of the two. The chiral
centers can be further designated as R or S or R,S or
d, D, 1, L or d, l, D, L.
15 Regarding the compounds and combinatorial
libraries described herein, the suffix "ene" added to any
of the described radical terms means that the radical is
connected to two parts of the compound (for example,
methylene (-CHI-) , ethylene (-CH~CH=-) , etc. ) .
The term "C1 to C12 alkyl" denotes such radicals
as methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-
butyl, sec-butyl, tart-butyl, amyl; tart-amyl, hexyl,
heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the
like. Preferred "C1 to C12 alkyl" groups are methyl,
ethyl, iso-butyl, sec-butyl and iso-propyl. Similarly,
the term "C1 to C1, alkylene" denotes radicals of 1 to 12
carbons connected to two other parts in the compound.
The term "C~ to C1? alkenyl" denotes such
radicals as vinyl, allyl, 2-butenyl, 3-butenyl, 2-
pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl,
4-hexenyl, 5-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl,
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5-heptenyl, 6-heptenyl, (as well as octenyl, nonenyl,
decenyl, undecenyl, dodecenyl radicals attached at any
appropriate carbon position and the like) as well as
dimes and trienes of straight and branched chains.
The term "Cz to Clz alkynyl" denotes such
radicals as ethanol, propynyl, 2-butynyl, 2-pentynyl, 3-
pentynyl, 2- hexynyl, 3-hexynyl, 4-hexynyl, 2-heptynyl,
3-heptynyl, 4- heptynyl, 5-heptynyl (as well as octynyl,
nonynyl, decynyl, undecynyl, dodecynyl radicals attached
l0 at any appropriate carbon position and the like) as well
as di- and tri-ynes of straight and branched chains.
The terms "C1 to Clz substituted alkyl, " "Cz to
C,z substituted alkenyl, " "Cz to C~, substituted alkynyl, "
"C1 to Clz substituted alkylene, " "Cz to Cl_ substituted
alkenylene" and "Cz to Clz substituted alkyny~ene" denote
groups are substituted by one or more, and preferably one
or two, halogen, hydroxy, protected hydroxy, oxo,
protected oxo, C3 to C, cycloalkyl, phenyl, naphthyl,
amino, protected amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino, guanidino,
protected guanidino, heterocyclic ring, substituted
heterocyclic ring, imidazolyl, indolyl, pyrrolidinyl, C1
to Clz alkoxy, C1 to Clz acyl, C1 to Clz acyloxy, nitro,
carboxy, protected carboxy, carbamoyl, carboxamide,
protected carboxamide, N-(C1 to C,z alkyl)carboxamide,
protected N- (C1 to Clz alkyl) carboxamide, N, N-di (C1 to Clz
alkyl)carboxamide, cyano, methylsulfonylamino, thiol, C1
to Clo alkylthio or C1 to Clo alkylsulfonyl groups. The
substituted alkyl groups may be substituted once or more,
and preferably once or twice, with the same or with
different substituents.
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Examples of the above substituted alkyl groups
include the 2-oxo-prop-1-yl, 3-oxo-but-1-yl, cyanomethyl,
nitromethyl, chloromethyl, hydroxymethyl,
tetrahydropyrar~yloxymethyl, trityloxymethyl,
propionyloxymethyl, aminomethyl, carboxymethyl,
allyloxycarbonylmethyl, allyloxycarbonylaminomethyl,
methoxymethyl, ethoxymethyl, t-butoxymethyl,
acetoxymethyl, chloromethyl, bromomethyl, iodomethyl,
trifluoromethyl, 6-hydroxyhexyl, 2,4-dichloro(n-butyl),
l0 2-aminopropyl, 1-chloroethyl, 2-chloroethyl, 1-
bromoethyl, 2-chloroethyl, 1-fluoroethyl, 2-fluoroethyl,
1- iodoethyl, 2-iodoethyl, 1-chloropropyl, 2-
chloropropyl, 3- chloropropyl, 1-bromopropyl, 2-
bromopropyl, 3-bromopropyl, 1-fluoropropyl, 2-
fluoropropyl, 3-fluoropropyl, 1- iodopropyl, 2-
iodopropyl, 3-iodopropyl, 2-aminoethyl, 1- aminoethyl, N-
benzoyl-2-aminoethyl, N-acetyl-2-aminoethyl, N-benzoyl-1-
aminoethyl, N-acetyl-1-aminoethyl and the like.
Examples of the above substituted alkenyl
groups include styrenyl, 3-chloro-propen-1-yl, 3-chloro-
buten-1-yl, 3-methoxy-propen-2-yl, 3-phenyl-buten-2-yl,
1-cyano-buten-3-yl and the like. The geometrical
isomerism is not critical, and all geometrical isomers
for a given substituted alkenyl can be used.
Examples of the above substituted alkynyl
groups include phenylacetylen-1-yl, 1-phenyl-2-propyn-1-
yl and the like.
The term "oxo" denotes a carbon atom bonded to
two additional carbon atoms substituted with an oxygen
atom doubly bonded to the carbon atom, thereby forming a
ketone moiety.
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The term "protected oxo" denotes a carbon atom
bonded to two additional carbon atoms substituted with
two alkoxy groups or twice bonded to a substituted diol
moiety, thereby forming an acyclic or cyclic ketal
moiety.
The term "C~ to C1, alkoxy" as used herein
denotes groups such as methoxy, ethoxy, n-propoxy,
isopropoxy, n-butoxy, t-butoxy and like groups. A
preferred alkoxy is methoxy. The term "C1 to C1,
substituted alkoxy" means the alkyl portion of the alkoxy
can be substituted in the same manner as in relation to C,
to C1~ substituted alkyl. Similarly, the term "C1 to C12
phenylalkoxy" as used herein means "C1 to C1= alkoxy"
bonded to a phenyl radical.
The term "C- to C1, acyloxy" denotes herein
groups such as formyloxy, acetoxy, propionyloxy,
butyryloxy, pivaloyloxy, pentanoyloxy, hexanoyloxy,
heptanoyloxy, octanoyloxy, nonanoyloxy, decanoyloxy,
undecanoyloxy, dodecanoyloxy and the like.
Similarly, the term "C1 to C12 acyl" encompasses
groups such as formyl, acetyl, propionyl, butyryl,
pentanoyl, pivaloyl, hexanoyl, heptanoyl, octanoyl,
nonanoyl, decanoyl, undecanoyl, dodecanoyl, benzoyl and
the like. Preferred acyl groups are acetyl and benzoyl.
The term "C1 to C1~ substituted acyl" denotes
the acyl group substituted by one or more, and preferably
one or two, halogen, hydroxy, protected hydroxy, oxo,
protected oxo, cyclohexyl, naphthyl, amino, protected
amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino, guanidino,
heterocyclic ring, substituted heterocyclic ring,
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imidazolyl, indolyl, pyrrolidinyl, C: to C1= alkoxy, C1 to
C1~ acyl, C, to C;, acyloxy, nitro, C: to C,; alkyl ester,
carboxy, protected carboxy, carbamoyl, carboxamide,
protected carboxamide, N-(C~ to C1, alkyl)carboxamide,
protected N- (C1 to C1~ alkyl) carboxamide, N, N-di (C1 to Cla
alkyl)carboxamide, cyano, methylsulfonylamino, thiol, C;
to C;:. alkylthio or C1 to C;o alkylsulfonyl groups. The
substituted aryl groups may be substituted once or more,
and preferably once or twice, with the same or with
different substituents.
Examples of C~ to Cla substituted acyl groups
include 4-phenylbutyroyl, 3-phenylbutyroyl,
3-phenylpropanoyl, 2- cyclohexanylacetyl,
cyclohexanecarbonyl, 2-furanoyl and
3-dimethylaminobenzoyl.
The substituent term "C_ to C~ cycloalkyl"
includes the cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl or cycloheptyl rings. Similarly, a
substituent that can be C3 to C, cycloalkyl" can also be
"CS to C, cycloalkyl," which includes the cyclopentyl,
cyclohexyl or cycloheptyl rings.
The substituent term "C3 to C~ substituted
cycloalkyl" or "CS to C~ substituted cycloalkyl" indicates
the above cycloalkyl rings substituted by one or two
halogen, hydroxy, protected hydroxy, C1 to Clo alkylthio,
C~ to C,o alkylsulfoxide, C, to Clo alkylsulfonyl, C1 to Clo
substituted alkylthio, C1 to Clo substituted
alkylsulfoxide, C1 to Clo substituted alkylsulfonyl, C1 to
C12 alkyl, C, to C1~ alkoxy, C1 to Cle substituted alkyl, C
to C1, alkoxy, oxo, protected oxo, (monosubstituted)amino,
(disubstituted)amino, trifluoromethyl, carboxy, protected
carboxy, phenyl, substituted phenyl, phenylthio,
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phenylsulfoxide, phenylsulfonyl, amino, or protected
amino groups.
The~term "cycloalkylene" means a cycloalkyl, as
defined above, where the cycloalkyl radical is bonded at
5 two positions connecting together two separate additional
groups. Similarly, the term "substituted cycloalkylene"
means a cycloalkylene where the cycloalkyl radical is
bonded at two positions connecting together two separate
additional groups and further bearing at least one
10 additional substituent.
The term "C~ to C; cycloalkenyl" indicates a
1,2, or 3-cyclopentenyl ring, a 1,2,3 or 4-cyclohexenyl
ring or a 1,2,3,4 or 5-cycloheptenyl ring, while the term
"substituted C~ to C, cycloalkenyl" denotes the above C,
15 to C., cycloalkenyl rings substituted by a C, to Cla alkyl
radical, halogen, hydroxy, protected hydroxy, C1 to Clz
alkoxy, trifluoromethyl, carboxy, protected carboxy, oxo,
protected oxo, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino, phenyl,
20 substituted phenyl, amino, or protected amino.
The term "CS to C, cycloalkenylene" is a
cycloalkenyl ring, as defined above, where the
cycloalkenyl radical is bonded at two positions
connecting together two separate additional groups.
Examples of CS to C7 cycloalkenylenes include
1,3-cyclopentylene and 1,2-cyclohexylene.
Similarly, the term "substituted C~ to C~
cycloalkenylene" means a cycloalkenylene further
substituted by halogen, hydroxy, protected hydroxy, C1 to
Clo alkylthio, C1 to Clo alkylsulfoxide, C1 to Clo
alkylsulfonyl, C1 to Clo substituted alkylthio, C, to C,o
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substituted alkylsulfoxide, C~ to C1~, substituted
alkylsulfonyl, C; to C,~ alkyl, CI to C~_ alkoxy, C1 to C1,
substituted alkyl, C, to C1, alkoxy, oxo, protected oxo,
(monosubstituted)amino, (disubstituted)amino,
trifluoromethyl, carboxy, protected carboxy, phenyl,
substituted phenyl, phenylthio, phenylsulfoxide,
phenylsulfonyl, amino, or protected amino group.
Examples of substituted C, to C, cycloalkenylenes include
4-chloro-1,3-cyclopentylene and
4-methyl-1,2-cyclohexylene.
The term "heterocycle" or "heterocyclic ring"
denotes optionally substituted five-membered to eight-
membered rings that have 1 to 4 heteroatoms, such as
oxygen, sulfur and/or nitrogen, in particular nitrogen,
either alone or in conjunction with sulfur or oxygen ring
atoms. These five-membered to eight-membered rings may
be saturated, fully unsaturated or partially unsaturated,
with fully saturated rings being preferred. Preferred
heterocyclic rings include morpholino, piperidinyl,
piperazinyl, 2-amino-imidazoyl, tetrahydrofurano,
pyrrolo, tetrahydrothiophen-yl, hexylmethyleneimino and
heptylmethyleneimino.
The term "substituted heterocycle" or
"substituted heterocyclic ring" means the above-described
heterocyclic ring is substituted with, for example, one
or more, and preferably one or two, substituents which
are the same or different which substituents can be
halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to
C1~ alkyl, C1 to C1~ alkoxy, C1 to C1~ substituted alkoxy, C1
to C,= acyl, C1 to C1~ acyloxy, carboxy, protected carboxy,
carboxymethyl, protected carboxymethyl, hydroxymethyl,
protected hydroxymethyl, amino, protected amino,
(monosubstituted)amino, protected (monosubstituted)amino,
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(disubstituted)amino carboxamide, protected carboxamide,
N- (C1 to C~~ alkyl) carboxamide, protected N- (C1 to Clz
alkyl) carboxamide, N, N-di (C1 to Clz alkyl) carboxamide,
trifluoromethyl, N- ( (C1 to Clz alkyl) sulfonyl) amino, N-
(phenylsulfonyl)amino, heterocycle or substituted
heterocycle groups.
The term "heteroaryl" means a heterocyclic
aromatic derivative which is a five-membered or six-
membered ring system having from 1 to 4 heteroatoms, such
as oxygen, sulfur and/or nitrogen, in particular
nitrogen, either alone or in conjunction with sulfur or
oxygen ring atoms. Examples of heteroaryls include .
pyridinyl, pyrimidinyl, and pyrazinyl, pyridazinyl,
pyrrolo, furano, oxazolo, isoxazolo, phthalimido,
thiazolo and the like.
The term "substituted heteroaryl" means the
above-described heteroaryl is substituted with, for
example, one or more, and preferably one or two,
substituents which are the same or different which
substituents can be halogen, hydroxy, protected~hydroxy,
cyano, nitro, C1 to Clz alkyl, C1 to Clz alkoxy, C1 to Clz
substituted alkoxy, C1 to Clz acyl, C~ to Ciz substituted
acyl, C1 to Clz acyloxy, carboxy, protected carboxy,
carboxymethyl, protected carboxymethyl, hydroxymethyl,
protected hydroxymethyl, amino, protected amino,
(monosubstituted)amino, protected (monosubstituted)amino,
(disubstituted)amino, carboxamide, protected carboxamide,
N- (C1 to Clz alkyl ) carboxamide, protected N- (C1 to Clz
alkyl)carboxamide, N, N-di(C1 to Clz alkyl)carboxamide,
trifluoromethyl, N-((C1 to Clz alkyl)sulfonyl)amino or N-
(phenylsulfonyl)amino groups.
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The term "C, to C1;; phenylalkyl" denotes a C1 to
C,Z alkyl group substituted at any position within the
alkyl chain by a phenyl. The definition includes groups
of the formula: -phenyl-alkyl, -alkyl-phenyl and -alkyl-
s phenyl-alkyl. Examples of such a group include benzyl,
2-phenylethyl, 3-phenyl(n-propyl), 4-phenylhexyl, 3-
phenyl(n-amyl), 3-phenyl(sec-butyl) and the like.
Preferred C~ to C1~ phenylalkyl groups are any one of the
preferred alkyl groups described herein combined witl-r-~.- '"
phenyl group.
Similarly, the term "C1 to Cla heterocycloalkyl"
denotes a C1 to C1~ alkyl group substituted at any
position within the alkyl chain by a ~~heterocycle," as
defined herein. The definition includes groups of the
formula: -heterocyclic-alkyl, -alkyl-heterocyclic and
-alkyl-heterocyclic-alkyl. Examples of such a group
include 2-pyridylethyl, 3-pierydyl(n-propyl), 4-
furylhexyl, 3-piperazyl(n-amyl), 3-morpholyl(sec-butyl)
and the like. Preferred C1 to C1~ heterocycloalkyl groups
are any one of the preferred alkyl groups described
herein combined with any one of the preferred heterocycle
groups described herein.
The terms "C, to CI8 substituted phenylalkyl"
and "C1 to C12 substituted heterocycloalkyl" denote a C~ to
C18 phenylalkyl group or C1 to C1~ heterocycloalkyl
substituted (on the alkyl or, where applicable, phenyl or
heterocyclic portion) with one or more, and preferably
one or two, groups chosen from halogen, hydroxy,
protected hydroxy, oxo, protected oxo, amino, protected
amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino, guanidino,
protected guanidino, heterocyclic ring, substituted
heterocyclic ring, C1 to C12 alkyl, C1 to Cle substituted
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alkyl, C1 to Clz alkoxy, C1 to Clz substituted alkoxy, C1 to
C1., aryl, C1 to Clz substituted acyl, C1 to C1, acyloxy,
nitro, carboxy, protected carboxy, carbamoyl,
carboxamide, protected carboxamide, N-(C, to C,z
alkyl)carboxamide, protected N-(C1 to Clz
alkyl) carboxamide, N, N-(C1 to Clz dialkyl) carboxamide,
cyano, N- (C1 to Clz alkylsulfonyl) amino, thiol, C1 to Clo
alkylthio, C1 to C1~, alkylsulfonyl groups; and/or the
phenyl group may be substituted with one or more, and
preferably one or two, substituents chosen from halogen,
hydroxy, protected hydroxy, cyano, nitro, C1 to Clz alkyl,
C1 to Clz substituted alkyl, C1 to Clz alkoxy, C~ to Clz
substituted alkoxy, C1 to C,z acyl, Cz to Clz substituted
acyl, C1 to Clz acyloxy, carboxy, protected carboxy,
carboxymethyl, protected carboxymethyl, hydroxymethyl,
protected hydroxymethyl, amino, protected amino,
(monosubstituted).amino, protected (monosubstituted)amino,
(disubstituted)amino, carboxamide, protected carboxamide,
N- (C, to Clz alkyl ) carboxamide, protected N- (C1 to Clz
alkyl)carboxamide, N, N-di(C1 to Clz alkyl)carboxamide,
trifluoromethyl, N- ( (C1 to Czz alkyl) sulfonyl) amino, N-
(phenylsulfonyl) amino, cyclic CZ to Clz alkylene or a
phenyl group, substituted or unsubstituted, for a
resulting biphenyl group. The substituted alkyl, phenyl
or heterocyclic groups may be substituted with one or
more, and preferably one or two, substituents which can
be the same or different.
Examples of the term "C~ to C1s substituted
phenylalkyl" include groups such as 2-phenyl-1-
chloroethyl, 2-(4-methoxyphenyl)ethyl, 4-(2,6-dihydroxy
phenyl)n-hexyl, 2-(5-cyano-3-methoxyphenyl)n-pentyl, 3-
(2,6-dimethylphenyl)n-propyl, 4-chloro-3-aminobenzyl, 6-
(4-methoxyphenyl)-3-carboxy(n-hexyl), 5-(4-
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aminomethylphenyl)- 3-(aminomethyl)n-pentyl, 5-phenyl-3-
oxo-n-pent-1-yl and the like.
The term "C, to C1~ phenylalkylene" specifies a
C, to C15 phenylalkyl, as defined above, where the
5 phenylalkyl radical is bonded at two different positions
connecting together two separate additional groups. The
definition includes groups of the formula:
-phenyl-alkyl-, -alkyl-phenyl- and -alkyl-phenyl-alkyl-.
Substitutions on the phenyl ring can be 1,2, 1,3 or 1,4.
20 C, to C1g phenylalkylenes include, for example,
1,4-toluylene and 1,3-xylylene.
Similarly, the term "C1 to C~,
heterocycloalkylene" specifies a C1 to C,n
heterocycloalkyl, as defined above, where the
l5 heterocycloalkyl radical is bonded at two different
positions connecting together two separate additional
groups. The definition includes groups of the formula:
-heterocyclic-alkyl-, -alkyl-heterocyclic and -alkyl-
heterocyclic-alkyl-.
20 The terms "C~ to C1$ substituted phenylalkylene"
and "C1 to C12 substituted heterocycloalkylene" means a C,
to C18 phenylalkylene or C1 to C12 heterocycloalkylene as
defined above that is further substituted by halogen,
hydroxy, protected hydroxy, C'1 to Clo alkylthio, C1 to Clo
25 alkylsulfoxide, C1 to Coo alkylsulfonyl, C1 to C1~
substituted alkylthio, C1 to Clo substituted
alkylsulfoxide, C1 to Clo substituted alkylsulfonyl, C1 to
C1, alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkyl, C,
to C1= alkoxy, oxo, protected oxo, (monosubstituted)amino,
(disubstituted)amino, trifluoromethyl, carboxy, protected
carboxy, phenyl, substituted phenyl, phenylthio,
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phenylsulfoxide, phenylsulfonyl, amino, or protected
amino group on the phenyl ring or on the alkyl group.
The term "substituted phenyl" specifies a
phenyl group substituted with one or more and preferably
one or two, moieties chosen from the groups consisting of
halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to
C1, alkyl, C1 to C1~ substituted alkyl, C1 to C_~ alkoxy, C1
to C1, substituted alkoxy, C1 to C1~ acyl, C1 to C1~
substituted acyl, C1 to C1~ acyloxy, carboxy, protected
carboxy, carboxymethyl, protected carboxymethyl,
hydroxymethyl, protected hydroxymethyl, amino, protected
amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino,
carboxamide, protected carboxamide, N-(C1 to C;,
alkyl) carboxamide, protected N- (C1 to C12
alkyl)carboxamide, N, N-di(C1 to C1, alkyl)carboxamide,
trifluoromethyl, N-((C1 to C" alkyl)sulfonyl)amino, N-
(phenylsulfonyl)amino or phenyl, wherein the phenyl is
substituted or unsubstituted, such that, for example, a
biphenyl results.
Examples of the term "substituted phenyl"
includes a mono- or di(halo)phenyl group such as 2, 3 or
4-chlorophenyl, ~,6-dichlorophenyl, 2,5-dichlorophenyl,
3,4-dichlorophenyl, 2, 3 or 4-bromophenyl,
3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2, 3 or
4-fluorophenyl and the like; a mono or di(hydroxy)phenyl
group such as 2, 3 or 4-hydroxyphenyl,
2,4-dihydroxyphenyl, the protected-hydroxy derivatives
thereof and the like; a nitrophenyl group such as 2, 3 or
4-nitrophenyl; a cyanophenyl group, for example, 2, 3 or
4-cyanophenyl; a mono- or di(alkyl)phenyl group such as
2, 3 or 4-methylphenyl, 2,4-dimethylphenyl, 2, 3 or
4-(iso-propyl)phenyl, 2, 3 or 4-ethylphenyl, 2, 3 or
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4-(n-propyl)phenyl and the like; a mono or
di(alkoxyl)phenyl group, for example,
2,6-dimethoxyphenyl, 2, 3 or 4-methoxyphenyl, 2, 3 or
4-ethoxyphenyl; 2, 3 or 4-(isopropoxy)phenyl, 2, 3 or
4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl and the
like; 2, 3 or 4-trifluoromethylphenyl; a mono- or
dicarboxyphenyl or (protected carboxy)phenyl group such
as 2, 3 or 4-carboxyphenyl or 2,4-di(protected
carboxy)phenyl; a mono-or di(hydroxymethyl)phenyl or
(protected hydroxymethyl)phenyl such as 2, 3, or
4-(protected hydroxymethyl)phenyl or
3,4-di(hydroxymethyl)phenyl; a mono- or
di(aminomethyl)phenyl or (protected aminomethyl)phenyl
such as 2, 3 or 4-(aminomethyl)phenyl or 2,4-(protected
aminomethyl)phenyl; or a mono- or
di(N-(methylsulfonylamino))phenyl such as 2, 3 or
4-(N-(methylsulfonylamino))phenyl. Also, the term
"substituted phenyl" represents disubstituted phenyl
groups wherein the substituents are different, for
example, 3-methyl-4-hydroxyphenyl, 3-chloro-4-
hydroxyphenyl, 2-methoxy-4-bromophenyl,
4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,
2-hydroxy 4-chlorophenyl and the like.
The term "phenoxy" denotes a phenyl bonded to
an oxygen atom, wherein the binding to the rest of the
molecule is through the oxygen atom. The term
"substituted phenoxy" specifies a phenoxy group
substituted with one or more, and preferably one or two,
moieties chosen from the groups consisting of halogen,
hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl,
C1 to C1z alkoxy, C1 to C12 substituted alkoxy, Cto C,
acyl, C1 to C1~ acyloxy, carboxy, protected carboxy,
carboxymethyl, protected carboxymethyl, hydroxymethyl,
protected hydroxymethyl, amino, protected amino,
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(monosubstituted)amino, protected (monosubstituted)amino,
(disubstituted)amino, carboxamide, protected carboxamide,
N- (C1 to C1~ alkyl ) carboxamide, protected N- (C1 to Clz
alkyl) carboxamide, N, N-di (C1 to C1z alkyl) carboxamide,
trifluoromethyl, N-((C1 to C1? alkyl)sulfonyl)amino and N-
(phenylsulfonyl)amino.
Examples of substituted phenoxy include
2-methylphenoxy, 2-ethylphenoxy, 2-propylphenoxy,
2-isopropylphenoxy, 2-sec-butylphenoxy,
2-tert-butylphenoxy, 2-allylphenoxy, 2-propenylphenoxy,
2-cyclopentylphenoxy, 2-fluorophenoxy,
2-(trifluoromethyl)phenoxy, 2-chlorophenoxy,
2-bromophenoxy, 2-methoxyphenoxy, 2-ethoxyphenoxy,
2-isopropoxyphenoxy, 3-methylphenoxy, 3-ethylphenoxy,
3-isopropylphenoxy, 3-tert-butylphenoxy,
3-pentadecylphenoxy, 3-(trifluoromethyl)phenoxy,
3-fluorophenoxy, 3-chlorophenoxy, 3-bromophenoxy,
3-iodophenoxy, 3-methoxyphenoxy,
3-(trifluoromethoxy)phenoxy, 4-methylphenoxy,
4-ethylphenoxy, 4-propylphenoxy, 4-isopropylphenoxy,
4-sec-butylphenoxy, 4-tert-butylphenoxy,
4-tert-amylphenoxy, 4-nonylphenoxy, 4-dodecylphenoxy,
4-cyclopenylphenoxy, 4-(trifluoromethyl)phenoxy,
4-fluorophenoxy, 4-chlorophenoxy, 4-bromophenoxy,
4-iodophenoxy, 4-methoxyphenoxy,
4-(trifluoromethoxy)phenoxy, 4-ethoxyphenoxy,
4-propoxyphenoxy, 4-butoxyphenoxy, 4-hexyloxyphenoxy,
4-heptyloxyphenoxy, 2,3-dimethylphenoxy,
5,6,7,8-tetrahydro-1-naphthoxy, 2,3-dichlorophenoxy,
2,3-dihydro-2,2-dimethyl-7-benzofuranoxy,
2,3-dimethoxyphenoxy, 2,6-dimethylphenoxy,
2,6-diisopropylphenoxy, 2,6-di-sec-butylphenoxy, 2-tert-
butyl-6-methylphenoxy, 2,6-di-tert-butylphenoxy, 2-allyl-
6-methylphenoxy, 2,6-difluorophenoxy,
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2,3-difluorophenoxy, 2,6-dichlorophenoxy,
2,6-dibromophenoxy, 2-fluoro-6-methoxyphenoxy,
2,6-dimethoxyphenoxy, 3,5-dimethylphenoxy, 5-isopropyl-
3-methylphenoxy, 3,5-di-tart-butylphenoxy,
3,5-bis(trifluoromethyl)phenoxy, 3,5-difluorophenoxy,
3,5-dichlorophenoxy, 3,5-dimethoxyphenoxy, 3-chloro-5-
methoxyphenoxy, 3,4-dimethylphenoxy, 5-indanoxy,
5,6,7,8-tetrahydro-2-naphthoxy, 4-chloro-3-methylphenoxy,
2,4-dimethylphenoxy, 2,5-dimethylphenoxy, 2-isopropyl-
5-methylphenoxy, 4-isopropyl-3-methylphenoxy,
5-isopropyl-2-methylphenoxy, 2-tart-butyl-
5-methylphenoxy, 2-tart-butyl-4-methylphenoxy,
2,4-di-tart-butylphenoxy, 2,4-di-tart-amylphenoxy,
4-fluoro-2-methylphenoxy, 4-fluoro-3-methylphenoxy,
2-chloro-4-methylphenoxy, 2-chloro-5-methylphenoxy,
4-chloro-2-methylphenoxy, 4-chloro-3-ethylphenoxy,
2-bromo-4-methylphenoxy, 4-iodo-2-methylphenoxy,
2-chloro-5-(trifluoromethyl)phenoxy, 2,4-difluorophenoxy,
2,5-difluorophenoxy, 3,4-difluorophenoxy, 4-chloro-2-
fluorophenoxy, 3-chloro-4-fluorophenoxy, 4-chloro-3-
fluorophenoxy, 2-bromo-4-fluorophenoxy, 4-bromo-2-
fluorophenoxy, 2-bromo-5-fluorophenoxy,
2,4-dichlorophenoxy, 3,4-dichlorophenoxy,
2,5-dichlorophenoxy, 2-bromo-4-chlorophenoxy, 2-chloro-4-
fluorophenoxy, 4-bromo-2-chlorophenoxy,
2,4-dibromophenoxy, 2-methoxy-4-methylphenoxy, 4-allyl-2-
methylphenoxy, trans-2-ethoxy-5-(1-propenyl)phenoxy,
2-methoxy-4-propenylphenoxy, 3,4-dimethoxyphenoxy,
3-ethoxy-4-methoxyphenoxy, 4-allyl-2,6-dimethoxyphenoxy,
3,4-methylenedioxyphenoxy, 2,3,6-trimethylphenoxy,
2,4-dichloro-3-methylphenoxy, 2,3,4-trifluorophenoxy,
2,3,6-trifluorophenoxy, 2,3,5-trifluorophenoxy,
2,3,4-trichlorophenoxy, 2,3,6-trichlorophenoxy,
2,3,5-trimethylphenoxy, 3,4,5-trimethylphenoxy, 4-chloro-
3,5-dimethylphenoxy. 4-bromo-3,5-dimethylphenoxy,
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2,4,6-trimethylphenoxy, 2,6-bis(hydroxymethyl)-4-
methylphenoxy, 2,6-di-tert-butyl-4-methylphenoxy, 2,6-
di-tent-butyl-4-methoxyphenoxy, 2,4,5- trifluorophenoxy,
2-chloro-3,5-difluorophenoxy, 2,4,6-trichlorophenoxy,
5 3,4,5-trimethoxyphenoxy, 2,3,5-trichlorophenoxy, 4-bromo-
2,6-dimethylphenoxy, 4-bromo-6-chloro-2-methylphenoxy,
2,6-dibromo-4-methylphenoxy, 2,6-dichloro-4-
fluorophenoxy, 2,~-dibromo-4-fluorophenoxy,
2,4,6-tribromophenoxy, 2,4,6-triiodophenoxy, 2-chloro-
10 4,5-dimethylphenoxy, 4-chloro-2-isopropyl-5-
methylphenoxy, 2-bromo-4,5-difluorophenoxy,
2,4,5-trichlorophenoxy, 2,3,5,6-tetrafluorophenoxy and
the like.
The term "C7 to C,~ substituted phenylalkoxy"
15 denotes a C-,-. to C1~ phenylalkoxy group bonded to the rest
of the molecule through the oxygen atom, wherein the
phenylalkyl portion is substituted with one or more, and
preferably one or two, groups selected from halogen,
hydroxy, protected hydroxy, oxo, protected oxo, amino,
20 protected amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino, guanidino,
heterocyclic ring, substituted heterocyclic ring, C1 to
C12 alkoxy, C1 to Cz2 acyl, C1 to C12 acyloxy, vitro,
carboxy, protected carboxy, carbamoyl, carboxamide,
25 protected carboxamide, N- (C1 to Clz alkyl) carboxamide,
protected N- (C1 to C1~ alkyl) carboxamide, N, N- (C1 to Cla
dialkyl ) carboxamide, cyano, N- (C1 to Clz
alkylsulfonyl)amino, thiol, Cl to Clo alkylthio, C1'to Clo
alkylsulfonyl groups; and/or the phenyl group can be
30 substituted with one or more, and preferably one or two,
substituents chosen from halogen, hydroxy, protected
hydroxy, cyano, vitro, C1 to C1z alkyl, C1 to C1, alkoxy,
C1 to Cla acyl, C1 to C1~ acyloxy, carboxy, protected
carboxy, carboxymethyl, protected carboxymethyl,
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hydroxymethyl, protected hydroxymethyl, amino, protected
amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino,
carboxamide, protected carboxamide, N-(CI to C:_ alkyl)
carboxamide, protected N-(C1 to C1? alkyl) carboxamide, N,
N-di (C1 to C1? alkyl) carboxamide, trifluoromethyl,
N- ( (C1 to C1= alkyl) sulfonyl) amino,
N-(phenylsulfonyl)amino or a phenyl group, substituted or
unsubstituted, for a resulting biphenyl group. The
substituted alkyl or phenyl groups may be substituted
with one or more, and preferably one or two, substituents
which can be the same or different.
Examples of the term "C~ to C1~ substituted
phenylalkoxy" include groups such as 2-(4-
hydroxyphenyl)ethoxy, 4-(4-methoxyphenyl)butoxy, (2R)-3-
phenyl-2-amino-propoxy, (2S)-3-phenyl-2-amino-propoxy,
2-indanoxy, 6-phenyl-1-hexanoxy, cinnamyloxy,
(+/-)-2-phenyl-1-propoxy, 2,2-dimethyl-3-phenyl-1-propoxy
and the like.
' The term "phthalimide" means a cyclic imide
which is made from phthalic acid, also called
1,2-benzenedicarboxylic acid. The term "substituted
phthalimide" specifies a phthalimide group substituted
with one or more, and preferably one or two, moieties
chosen from the groups consisting of halogen, hydroxy,
protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12
alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to
C12 acyloxy, carboxy, protected carboxy, carboxymethyl,
protected carboxymethyl, hydroxymethyl, protected
hydroxymethyl, amino, protected amino,
(monosubstituted)amino, protected (monosubstituted)amino,
(disubstituted)amino, carboxamide, protected carboxamide,
N- (C1 to C12 alkyl) carboxamide, protected N- (C: to CI,
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alkyl) carboxamide, N, _N-di (C1 to C12 alkyl) carboxamide,
trifluoromethyl, N- (°'(Ci to C1~ alkyl) sulfonyl) amino and
N-(phenylsulfonyl)amino.
Examples of substituted phthalimides include
4,5-dichlorophthalimido, 3-fluorophthalimido,
4-methoxyphthalimido, 3-methylphthalimido,
4-carboxyphthalimido and the like.
The term "substituted naphthyl" specifies a
naphthyl group substituted with one or more, and
preferably one or two, moieties either on the same ring
or on different rings chosen from the groups consisting
of halogen, hydroxy, protected hydroxy, cyano, nitro,
Ci to C6 alkyl, C1 to C., alkoxy, C1 to C, acyl, C1 to C,
acyloxy, carboxy, protected carboxy, carboxymethyl,
protected carboxymethyl, hydroxymethyl, protected
hydroxymethyl, amino, protected amino,
(monosubstituted)amino, protected (monosubstituted)amino,
(disubstituted)amino, carboxamide, protected carboxamide,
N- (C1 to C1~ alkyl) carboxamide, protected N- (C1 to Cla
alkyl)carboxamide, N, N-di(C~ to Clz alkyl)carboxamide,
trifluoromethyl, N- ( (C1 to C12 alkyl) sulfonyl) amino or
N-(phenylsulfonyl)amino.
Examples of the term "substituted naphthyl"
includes a mono or di(halo)naphthyl group such as 1, 2,
3, 4, 5, 6, 7 or 8-chloronaphthyl, 2, 6-dichloronaphthyl,
2, 5-dichloronaphthyl, 3, 4-dichloronaphthyl, 1, 2, 3, 4,
5, 6, 7 or 8-bromonaphthyl, 3, 4-dibromonaphthyl,
3-chloro-4-fluoronaphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-fluoronaphthyl and the like; a mono or
di(hydroxy)naphthyl group such as 1, 2, 3, 4, 5, 6, 7 or
8-hydroxynaphthyl, 2, 4-dihydroxynaphthyl, the protected-
hydroxy derivatives thereof and the like; a nitronaphthyl
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group such as 3- or 4-nitronaphthyl; a cyanonaphthyl
group, for example, l, 2, 3, 4, 5, 6, 7 or
8-cyanonaphthyl; a mono- or di(alkyl)naphthyl group such
as 2, 3, 4, 5,~6, 7 or 8-methylnaphthyl, l, 2,
4-dimethylnaphthyl, l, 2, 3, 4, 5, 6, 7 or
8-(isopropyl)naphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-ethylnaphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-(n-propyl)naphthyl and the like; a mono or
di(alkoxy)naphthyl group, for example, 2,
6-dimethoxynaphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-methoxynaphthyl, l, 2, 3, 4, 5, 6, 7 or
8-ethoxynaphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-(isopropoxy)naphthyl, 1, 2, 3, 4, 5, 6, 7 or
8-(t-butoxy)naphthyl, 3-ethoxy-4-methoxynaphthyl and the
like; l, 2, 3, 4, 5, 6, 7 or 8-trifluoromethylnaphthyl; a
mono- or dicarboxynaphthyl or (protected carboxy)naphthyl
group such as 1, 2, 3, 4, 5, 6, 7 or 8-carboxynaphthyl or
2, 4-di(-protected carboxy)naphthyl; a mono-or
di(hydroxymethyl)naphthyl or (protected
hydroxymethyl)naphthyl such as 1, 2, 3, 4, 5, 6, 7 or
8-(protected hydroxymethy.l)naphthyl or 3,
4-di(hydroxymethyl)naphthyl; a mono- or di(amino)naphthyl
or (protected amino)naphthyl such as 1, 2, 3, 9, 5, 6, 7
or 8-(amino)naphthyl or 2, 4-(protected amino)-naphthyl,
a mono- or di(aminomethyl)naphthyl or (protected
aminomethyl)naphthyl such as 2, 3, or
4-(aminomethyl)naphthyl or 2, 4-(protected aminomethyl)-
naphthyl; or a mono- or di-(N-methylsulfonylamino)
naphthyl such as l, 2, 3, 4, 5, 6, 7 or
8-(N-methylsulfonylamino)naphthyl. Also, the term
"substituted naphthyl" represents disubstituted naphthyl
groups wherein the substituents are different, for
example, 3-methyl-4-hydroxynaphth-1-yl, 3-chloro-9-
hydroxynaphth-2-yl, 2-methoxy-4-bromonaphth-1-yl,
4-ethyl-2-hydroxynaphth-1-yl, 3-hydroxy-9-nitronaphth-2-
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y1, 2-hydroxy-4-chloronaphth-1-yl, 2-methoxy-7-
bromonaphth-1-yl, 4-ethyl-5-hydroxynaphth-2-yl,
3-hydroxy-8-nitronaphth-2-yl, 2-hydroxy-5-chloronaphth-1-
yl and the like.
The term "naphthylene" means a naphthyl radical
bonded at two positions connecting together two separate
additional groups. Similarly, the term "substituted
napthylene" means a naphthylene group that is further
substituted by halogen, hydroxy, protected hydroxy, C1 to
Clo alkylthio, C1 to Clo alkylsulfoxide, C1 to Clo
alkylsulfonyl, . C1 to C,o substituted alkylthio, C1 to Clo
substituted alkylsulfoxide, Cl to Clo substituted
alkylsulfonyl, C1 to C1? alkyl, C~ to C1, alkoxy, C1 to Cla
substituted alkyl, C1 to C1z alkoxy, oxo, protected oxo,
(monosubstituted)amino, (disubstituted)amino,
trifluoromethyl, carboxy, protected carboxy, phenyl,
substituted phenyl, phenylthio, phenylsulfoxide,
phenylsulfonyl, amino, or protected amino group.
The terms "halo" and "halogen" refer to the
fluor~, chloro, bromo or iodo atoms. There can be one or
more halogens, which are the same or different.
Preferred halogens are chloro and fluoro.
The term "(monosubstituted)amino" refers to an
amino group with one substituent chosen from the group
consisting of phenyl, substituted phenyl, C1 to C1z alkyl,
C1 to C12 substituted alkyl, C1 to C12 aryl, C1 to C1~
substituted acyl, C~ to C1~ alkenyl, Cz to C1z substituted
alkenyl, CZ to Clz alkynyl, Cz to C1~ substituted alkynyl,
C, to C19 phenylalkyl, C, to Clg substituted phenylalkyl,
heterocyclic ring, substituted heterocyclic ring, Cl to
C1, heterocycloalkyl and C1 to C1z substituted
heterocycloalkyl. The (monosubstituted)amino can
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additionally have an amino-protecting group as
encompassed by the term "protected
(monosubstituted)amino."
The term "(disubstituted)amino" refers to an
5 amino group with two substituents chosen from the group
consisting of phenyl, substituted phenyl, C~ to C1z alkyl,
C1 to C1, substituted alkyl, C1 to C1~ acyl, CZ to C12
alkenyl, C~ to C1, alkynyl, C, to C18 phenylalkyl, C~ to C1~
substituted phenylalkyl, CI to Ciz heterocycloalkyl and C1
10 to C1~ substituted heterocycloalkyl,. The two
substituents can be the same or different.
The term "amino-protecting group" as used
herein refers to substituents of the amino group commonly
employed to block or protect the amino functionality
15 while reacting other functional groups of the molecule.
The term "protected (monosubstituted)amino" means there
is an amino-protecting group on the monosubstituted amino
nitrogen atom. In addition, the term "protected
earboxamide" means there is an amino-protecting group on
20 the carboxamide nitrogen. Similarly, the term "protected
N-(C1 to C1~ alkyl)carboxamide" means there is an amino-
protecting group on the carboxamide nitrogen.
Examples of such amino-protecting groups
include the formyl ("For") group, the trityl group, the
25 phthalimido group, the trichloroacetyl group, the
chloroacetyl, bromoacetyl, and iodoacetyl groups,
urethane-type blocking groups, such as t-butoxycarbonyl
("Boc"), 2-(4-biphenylyl)propyl-~-oxycarbonyl ("Bpoc"),
2-phenylpropyl-2-oxycarbonyl ("Poc"),
30 2-(4-xenyl)isopropoxycarbonyl, 1,1-diphenylethyl-1-
oxycarbonyl, 1,1-diphenylpropyl-1-oxycarbonyl, 2-(3,5-
dimethoxyphenyl)propyl-2-oxycarbonyl ("Ddz"), 2-(p-
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toluyl)propyl-2-oxycarbonyl, cyclopentanyloxycarbonyl,
1-methylcyclopentanyloxycarbonyl, cyclohexanyloxy-
carbonyl, 1-methylcyclohexanyloxycarbonyl,
2-methylcyclohexanyloxycarbonyl, 2-(4-toluylsulfonyl)-
ethoxycarbonyl, 2-(methylsulfonyl)ethoxycarbonyl,
2-(triphenylphosphino)-ethoxycarbonyl,
9-fluorenylmethoxycarbonyl ("Fmoc"),
2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl,
1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl,
5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyl-
oxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-
propoxycarbonyl, cyclopropylmethoxycarbonyl,
isobornyloxycarbonyl, 1-piperidyloxycarbonyl,
benzyloxycarbonyl '("Cbz"), 4-phenylbenzyloxycarbonyl,
2-methylbenzyloxy-carbonyl, -2,4,5,-
tetramethylbenzyloxycarbonyl ("Tmz"),
4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl,
4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,
2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyl-
oxycarbonyl, 4-bromobenzyloxycarbonyl,
3-bromobenzyloxycarbonyl, 4-nitrobenzyloxy-carbonyl,
4-cyanobenzyloxycarbonyl, 4-(decyloxy)benzyloxycarbonyl
and the like; the benzoylmethylsulfonyl group,
dithiasuccinoyl ("Dts"), the 2-(nitro)phenylsulfenyl
group ("Nps"), the diphenyl-phosphine oxide group and
like amino-protecting groups. The species of amino-
proteeting group employed is not critical so long as the
derivatized amino group is stable to the conditions of
the subsequent reactions) and can be removed at the
appropriate point without disrupting the remainder of the
compounds. Preferred amino-protecting groups are Boc,
Cbz and Fmoc. Further examples of amino-protecting
groups embraced by the above term are well known in
organic synthesis and the peptide art and are described
by, for example, T.W. Greene and P.G.M. Wuts, "Protective
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Groups in Organic Synthesis," 2nd ed., John Wiley and
Sons, New York, NY, 1991, Chapter 7, M. Bodanzsky,
"Principles of Peptide Synthesis," 1st and 2nd revised
ed., Springer-Verlag, New York, NY, 1984 and 1993, and
Stewart and Young, "Solid Phase Peptide Synthesis," 2nd
ed., Pierce Chemical Co., Rockford, IL, 1984, each of
which is incorporated herein by reference. The related
term "protected amino" defines an amino group substituted
with an amino-protecting group discussed above.
The term "protected guanidino" as used herein
refers to an "amino-protecting group" on one or two of
the guanidino nitrogen atoms. Examples of "protected
guanidino" groups are described by T.W. Greene and P.G.M.
Wuts; M. Bodanzsky; and Stewart and Young, supra.
The term "carboxy-protecting group" as used
herein refers to one of the ester derivatives of the
carboxylic acid group commonly employed to block or
protect the carboxylic acid group while reactions are
carried out on other functional groups on the compound.
Examples of such carboxylic acid protecting groups
include t-butyl, 4-nitrobenzyl, 4-methoxybenzyl,
3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl,
2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl,
pentamethylbenzyl, 3,4-methylenedioxybenzyl, benzhydryl,
4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl,
2-phenylpropyl, trimethylsilyl, t-butyldimethylsilyl,
phenacyl, 2,2,2-trichloroethyl, (trimethylsilyl)ethyl,
(di(n-butyl)methylsilyl)ethyl, p- toluenesulfonylethyl,
4-nitrobenzylsulfonylethyl, allyl, cinnamyl,
1-(trimethylsilylmethyl)propenyl and like moieties. The
species of carboxy-protecting group employed is not
critical so long as the derivatized carboxylic acid is
stable to the conditions of subsequent reactions) and
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can be removed at the appropriate point without
disrupting the remainder of the molecule. Further
examples of these groups are found in E. Haslam,
"Protective Groups in Organic Chemistry," J.G.W. McOmie,
Ed., Plenum Press, New York, NY, 2973, Chapter 5, and
T.W. Greene and P.G.M. Wuts, "Protective Groups in
Organic Synthesis," 2nd ed., John Wiley and Sons, New
York, NY, 1991, Chapter 5, each of which is incorporated
herein by reference. A related term is "protected
carboxy," which refers to a carboxy group substituted
with one of the above carboxy-protecting groups.
The term "hydroxy-protecting group" refers to
readily cleavable groups bonded to hydroxyl groups, such
as the tetrahydropyranyl, 2-methoxypropyl, 1-ethoxyethyl,
Z5 methoxymethyl, 2-methoxyethoxymethyl, methylthiomethyl,
t-butyl, t-amyl, trityl, 4-methoxytrityl,
4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl, benzyl,
allyl, trimethylsilyl, (t-butyl)dimethylsilyl,
2,2,2-trichloroethoxycarbonyl groups and the like. The
species of hydroxy-protecting groups is not critioal so
long as the derivatized hydroxyl group is stable to the
conditions of subsequent reactions) and can be removed
at the appropriate point without disrupting the remainder
of the molecule. Further examples of hydroxy-protecting
groups are described by C.B. Reese and E. Haslam,
"Protective Groups in Organic Chemistry," J.G.W. McOmie,
Ed., Plenum Press, New York, NY, 1973, Chapters 3 and 4,
respectively, and T.W. Greene and P.G.M. Wuts,
"Protective Groups in Organic Synthesis," 2nd ed., John
Wiley and Sons, New York, NY, 1991, Chapters 2 and 3.
Related terms are "protected hydroxy," and "protected
hydroxymethyl" which refer to a hydroxy or hydroxymethyl
substituted with one of the above hydroxy-protecting
groups.
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The term "C, to C,V alkylthio" refers to sulfide
groups such as methylthio, ethylthio, n-propylthio,
isopropylthio, n-butylthio, t-butylthio and like groups.
The term "C1 to C,o alkylsulfoxide" indicates sulfoxide
groups such as methylsulfoxide, ethylsulfoxide, n-
propylsulfoxide, isopropylsulfoxide, n-butylsulfoxide,
sec-butylsulfoxide and the like. The term "Ci to Clo
alkylsulfonyl" encompasses groups such as methylsulfonyl,
ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, n-
butylsulfonyl, t-butylsulfonyl and the like. it should
also be understood that the above thio, sulfoxide or
sulfonyl groups can be at any point on the alkyl chain
(e. g., 2-methylmercaptoethyl).
The terms "C_ to Clo substituted alkylthio,"
Z5 "C1 to Cio substituted alkylsulfoxide, " and "C~ to Clo
substituted alkylsulfonyl," denote the C, to C:~ alkyl
portion of these groups may be substituted as described
above in relation to "substituted alkyl."
The terms "phenylthio," "phenylsulfoxide," and
"phenylsulfonyl" specify a thiol, a sulfoxide, or
sulfone, respectively, containing a phenyl group. The
terms "substituted phenylthio," "substituted
phenylsulfoxide," and "substituted phenylsulfonyl" means
that the phenyl of these groups can be substituted as
described above in relation to "substituted phenyl."
The term "C1 to C1~ alkylaminocarbonyl" means a
C1 to C1~ alkyl attached to a nitrogen of the
aminocarbonyl group. Examples of C1 to C1,
alkylaminocarbonyl include methylaminocarbonyl,
ethylaminocarbonyl, propylaminocarbonyl and
butylaminocarbonyl. The term "C1 to C" substituted
alkylaminocarbonyl" denotes a substituted alkyl bonded to
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a nitrogen of the aminocarbonyl group, which alkyl may be
substituted as described above in relation to C1 to C,
substituted alkyl. Examples of C1 to C1~ substituted
alkylaminocarbonyl include, for example,
5 methoxymethylaminocarbonyl, 2-chloroethylaminocarbonyl,
2-oxopropylaminocarbonyl and 4-phenylbutylaminocarbonyl.
The term "C1 to C1~ alkoxycarbonyl" means a
"C1 to C1~ alkoxy" group attached to a carbonyl group.
The term "C1 to C1z substituted alkoxycarbonyl" denotes a
10 substituted alkoxy bonded to the carbonyl group, which
alkoxy may be substituted as described above in relation
to "C1 to C1~ substituted alkyl."
The term "phenylaminocarbonyl" means a phenyl
15 attached to a nitrogen of the aminocarbonyl group. The
term "substituted phenylaminocarbonyl" denotes a
substituted phenyl bonded to a nitrogen of the
aminocarbonyl group, which phenyl may be substituted as
described above in relation to substituted phenyl.
20 Examples of substituted phenylaminocarbonyl include
2-chlorophenylaminocarbonyl, 3-chlorophenylaminocarbonyl
2-nitorphenylaminocarbonyl, 4-biphenylaminocarbonyl,
and 4-methoxyphenylaminocarbonyl.
The term "C1 to C12 alkylaminothiocarbonyl"
25 means a C1 to Cla alkyl attached to an aminothiocarbonyl
group, wherein the alkyl has the same meaning as defined
above. Examples of C1 to Clz alkylaminothiocarbonyl
include methylaminothiocarbonyl, ethylaminothiocarbonyl,
propylaminothiocarbonyl and butylaminothiocarbonyl.
30 The term "C1 to C1~ substituted
alkylaminothiocarbonyl" denotes a substituted alkyl
bonded to an aminothiocarbonyl group, wherein the alkyl
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may be substituted as described above in relation to C1 to
C1, substituted alkyl. Examples of C1 to C1~ substituted
alkylaminothiocarbonyl include, for example,
methoxymethylaminothiocarbonyl,
2-chloroethylaminothiocarbonyl,
2-oxopropylaminothiocarbonyl and
4-phenylbutylaminothiocarbonyl.
The term "phenylaminothiocarbonyl" means a
phenyl attached to an aminothiocarbonyl group, wherein
the phenyl has the same meaning as defined above.
The term "substituted phenylaminothiocarbonyl"
denotes a substituted phenyl bonded to an
aminothiocarbonyl group, wherein phenyl may be
substituted as described above in relation to substituted
phenyl.. Examples of substituted phenylaminothiocarbonyls
include 2-chlorophenylaminothiocarbonyl,
3-chlorophenylaminothiocarbonyl,
2-nitorphenylaminothiocarbonyl,
4-biphenylaminothiocarbonyl and
4-methoxyphenylaminothiocarbonyl.
The term "phenylene" means a phenyl group where
the phenyl radical is bonded at two positions connecting
together two separate additional groups. Examples of
"phenylene" include 1,~-phenylene, 1,3-phenylene, and
1,4-phenylene.
The term "substituted phenylene" means a phenyl
group where the phenyl radical is bonded at two positions
connecting together two separate additional groups,
wherein the phenyl is substituted as described above in
relation to "substituted phenyl."
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The term "substituted Ci to C1~ alkylene" means
a C1 to C1, alkyl group where the alkyl radical is
connected to two additional groups and further bearing
(i.e., substituted with) an additional substituent.
Examples of a "substituted C1 to C12 alkylene" include
aminomethylene, 1-(amino)-1,2-ethylene, 2-(amino)-1,2-
ethylene, 1-(acetamido)-1,2-ethylene, 2-(acetamido)-1,2-
ethylene, 2-hydroxy-1,1-ethylene and 1-(amino)-1,3-
propylene.
The terms "cyclic CZ to C~ alkylene,"
"substituted cyclic Cz to C-; alkylene, " "cyclic C, to C,
heteroalkylene," and "substituted cyclic C~ to C~
heteroalkylene," defines such a cyclic group bonded
("fused") to the phenyl radical resulting in a bicyclic
ring system. The cyclic group may be saturated or
contain one or two double bonds. Furthermore, the cyclic
group may have one or two methylene or methine groups
replaced by one or two oxygen, nitrogen or sulfur atoms
which are the cyclic CZ to C~ heteroalkylene.
The cyclic alkylene or heteroalkylene group may
be substituted once or twice by the same or different
substituents which, if appropriate, can be connected to
another part of the compound (e. g., alkylene) selected
from the group consisting of the following moieties:
hydroxy, protected hydroxy, carboxy, protected carboxy,
oxo, protected oxo, C1 to Cq acyloxy, formyl, C1 to C1~
acyl, C1 to C12 alkyl, C, to C, alkoxy, C1 to Clo alkylthio,
C1 to Clo alkylsulfoxide, C1 to Coo alkylsulfonyl, halo,
amino, protected amino, (monosubstituted)amino, protected
(monosubstituted)amino, (disubstituted)amino,
hydroxymethyl or a protected hydroxymethyl.
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The cyclic alkylene or heteroalkylene group
fused to the benzene radical can contain two to ten ring
members and, preferably, contains three to six members.
Examples of such saturated cyclic groups are when the
resulting bicyclic ring system is a 2,3-dihydro-indanyl
or tetralin ring. When the cyclic groups are
unsaturated, the resulting bicyclic ring system can be
naphthyl or indolyl. Examples of fused cyclic groups
which contain one nitrogen atom and one or more double
bonds, preferably one or two double bonds, are when the
benzene radical is fused to a pyridino, pyrano, pyrrolo,
pyridinyl, dihydropyrrolo, or dihydropyridinyl ring.
Examples of fused cyclic groups which contain one oxygen
atom and one or two'double bonds are when the benzene
l5 radical ring is fused to a furo, pyrano, dihydrofurano,
or dihydropyrano ring. Examples of fused cyclic groups
which have one sulfur atom and contain one or two double
bonds are when the benzene radical is fused to a thieno,
thiopyrano, dihydrothieno or dihydrothiopyrano ring.
Examples of cyclic groups which contain two heteroatoms
selected from sulfur and nitrogen and one or two double
bonds are when the benzene radical ring is fused to a
thiazolo, isothiazolo, dihydrothiazolo or
dihydroisothiazolo ring. Examples of cyclic groups which
contain two heteroatoms selected from oxygen and nitrogen
and one or two double bonds are when the benzene ring is
fused to an oxazolo, isoxazolo, dihydrooxazolo or
dihydroisoxazolo ring. Examples of cyclic groups which
contain two nitrogen heteroatoms and one or two double
bonds occur when the benzene ring is fused to a pyrazolo,
imidazolo, dihydropyrazolo or dihydroimidazolo ring or
pyrazinyl.
One or more of the compounds of the invention,
even within a given library, may be present as a salt.
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The term "salt" encompasses those salts that form with
the carboxylate anions and amine nitrogens and include
salts formed with the organic and inorganic anions and
cations discussed below. Furthermore, the term includes
salts that form by standard acid-base reactions with
basic groups (such as amino groups) and organic or
inorganic acids. Such acids include hydrochloric,
hydrofluoric, trifluoroacetic, sulfuric, phosphoric,
acetic, succinic, citric, lactic, malefic, fumaric,
palmitic, cholic, pamoic, mucic, D-glutamic, D-camphoric,
glutaric, phthalic, tartaric, lauric, stearic,
salicyclic, methanesulfonic, benzenesulfonic, sorbic,
picric, benzoic, cinnamic, and like acids.
The term "organic or inorganic cation" refers
to counter-ions for the carboxylate anion of a
carboxylate salt. The counter-ions are chosen from the
alkali and alkaline earth metals, (such as lithium,
sodium, potassium, barium, aluminum and calcium);
ammonium and mono-, di- and tri-alkyl amines such as
trimethylamine, cyclohexylamine; and the organic cations,
such as dibenzylammonium, benzylammonium,
2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium,
phenylethylbenzylammonium, dibenzylethylenediammonium,
and like canons. See, for example, "Pharmaceutical
Salts," Berge et al., J. Pharm. Sci., 66:1-19 (1977),
which is incorporated herein by reference. Other cations
encompassed by the above term include the protonated form
of procaine, quinine and N-methylglucosamine, and the
protonated forms of basic amino acids such as glycine,
ornithine, histidine, phenylglycine, lysine and arginine.
Furthermore, any zwitterionic form of the instant
compounds formed by a carboxylic acid and an amino group
is referred to by this term. For example, a cation for a
carboxylate anion will exist when a position is
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substituted with a (quaternary ammonium)methyl group. A
preferred cation for the carboxylate anion is the sodium
cation.
The compounds of the invention can also exist
5 as solvates and hydrates. Thus, these compounds may
crystallize with, for example, waters of hydration, or
one, a number of, or any fraction thereof of molecules of
the mother liquor solvent. The solvates and hydrates of
such compounds are included within the scope of this
10 invention.
One or more compounds of the invention, even
when in a library, can be in the biologically active
ester form, such as the non-toxic, metabolically-labile
ester-form. Such ester forms induce increased blood
15 levels and prolong the efficacy of the corresponding non-
esterified forms of the compounds. Ester groups which
can be used include the lower alkoxymethyl groups, for
example, methoxymethyl, ethoxymethyl, isopropoxymethyl
and the like; the -(C1 to C1~) alkoxyethyl groups, for
20 example methoxyethyl, ethoxyethyl, propoxyethyl,
isopropoxyethyl and the like; the 2-oxo-1,3-diooxlen-4-
ylmethyl groups, such as 5-methyl-2-oxo-1,3-dioxolen-4-
ylmethyl, 5-phenyl-2-oxo-1,3-dioxolen-4-ylmethyl and the
like; the C1 to Clo alkylthiomethyl groups, for example
25 methylthiomethyl, ethylthiomethyl, iso-propylthiomethyl
and the like; the acyloxymethyl groups, for example
pivaloyloxymethyl, pivaloyloxyethyl, -acetoxymethyl and
the like; the ethoxycarbonyl-1-methyl group; the
-acetoxyethyl; the 1-(C1 to ClZ alkyloxycarbonyloxy)ethyl
30 groups such as the 1-(ethoxycarbonyloxy)ethyl group; and
the 1-(C1 to C1, alkylaminocarbonyloxy)ethyl groups such
as the 1-(methylaminocarbonyloxy)ethyl group.
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The term "amino acid" includes any one of the
twenty naturally-occurring amino acids or the D-form of
any one of the naturally-occurring amino acids. In
addition, thelterm "amino acid" also includes other non-
naturally occurring amino acids besides the D-amino
acids, which are functional equivalents of the naturally-
occurring amino acids. Such non-naturally-occurring
amino acids include, for example, norleucine ("Nle"),
norvaline ("Nva"), L- or D- naphthalanine, ornithine
("Orn"), homoarginine (homoArg) and others well known in
the peptide art, such as those described in M. Bodanzsky,
"Principles of Peptide Synthesis," 1st and 2nd revised
ed., Springer-Verlag, New York, NY, 1984 and 1993, and
Stewart and Young,."Solid Phase Peptide Synthesis," 2nd
ed., Pierce Chemical Co., Rockford, IL, 1984, both of
which are incorporated herein by reference. Amino acids
and amino acid analogs can be purchased commercially
(Sigma Chemical Co.; Advanced Chemtech) or synthesized
using methods known in the art.
The term "functionalized resin" means any
resin, crosslinked or otherwise, where functional groups
have been introduced into the resin, as is common in the
art. Such resins include, for example,~ those
functionalized with amino, alkylhalo, formyl or hydroxy
groups. Such resins which can serve as solid supports
are well known in the art and include, for example,
4-methylbenzhydrylamine-copoly(styrene-to divinylbenzene)
(MBHA), 4-hydroxymethylphenoxymethyl-copoly(styrene-1o
divinylbenzene), 4-oxymethyl-phenyl-acetamido-
copoly(stryene-1o divinylbenzene)(Wang), 4-(oxymethyl)-
phenylacetamido methyl (Pam), and TentagelT'~, from Rapp
Polymere Gmbh, trialkoxy-diphenyl-methyl ester-
copoly(styrene-1o divinylbenzene)(RINK) all of which are
commercially available. Other functionalized resins are
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known in the art and can be use without departure from
the scope of the current invention. Such resins may
include those described in Jung, G., Combinatorial
Peptide and Nonpeptide Libraries, A Handbook (VCH Verlag,
1996) or Bunin, B. A., The Combinatorial Index (Academic
Press, 1998) and are incorporated herein by reference.
As used herein, a "combinatorial library" is an
intentionally created collection of differing molecules
which can be prepared by the means provided below or
otherwise and screened for biological activity in a
variety of formats (e. g., libraries of soluble molecules,
libraries of compounds attached to resin beads, silica
chips or other solid supports). A "combinatorial
library," as defined above, involves successive rounds of
chemical syntheses based on a common starting structure.
The combinatorial libraries can be screened in any
variety of assays, such as those detailed below as well
as others useful for assessing their biological activity.
The combinatorial libraries will generally have at least
one active compound and are generally prepared such that
the compounds are in equimolar quantities.
Compounds described in previous work that are
not taught as part of a collection of compounds or not
taught as intended for use as part of such a collection
are not part of a "combinatorial library" of the
invention. In addition, compounds that are in an
unintentional or undesired mixture are not part of a
"combinatorial library" of the invention.
A combinatorial library of the invention can
contain two or more of the above-described bicyclic
hydantoin compounds. The invention further provides a
combinatorial library containing three, four or five or
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48
more of the above-described compounds. In another
embodiment of the invention, a combinatorial library can
contain ten or more of the above-described compounds. In
yet another embodiment of the invention, a combinatorial
library can contain fifty or more of the above-described
compounds. If desired, a combinatorial library of the
invention can contain 100,000 or more, or even 1,000,000
or more, of the above-described compounds. ,
By way of example, the preparation of the
combinatorial libraries can use the "split resin
approach." The split resin approach is described by, for
example, U.S. Patent 5,010,175 to Rutter, WO PCT 91/19735
to Simon, and Gallop et al., J. Med. Chem., 37:1233-1251
(1-994), all of which are incorporated herein by
reference.
The amino acids are indicated herein by either
their full name or by the commonly known three letter
code. Further, in the naming of amino acids, "D-"
designates an amino acid having the "D" configuration, as
opposed to the naturally occurring L-amino acids. Where
no specific configuration is indicated, one skilled in
the art would understand the amino acid to be an L-amino
acid. The amino acids can, however, also be in racemic
mixtures of the D- and L-configuration or the D-amino
acid can readily be substituted for that in the
L-configuration.
For preparing pharmaceutical compositions
containing compounds of the invention, inert,
pharmaceutically acceptable carriers are used. The
pharmaceutical carrier can be either solid or liquid.
Solid form preparations include, for example, powders,
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tablets, dispersible granules, capsules, cachets, and
suppositories.
A solid carrier can be one or more substances
which can also act as diluents, flavoring agents,
solubilizers, lubricants, suspending agents, binders, or
tablet disintegrating agents; it can also be an
encapsulating material.
In powders, the carrier is generally a finely
divided solid which is in a mixture with the finely
20 divided active component. In tablets, the active
compound is mixed with the carrier having the necessary
binding properties in suitable proportions and compacted
in the shape and size desired.
For preparing pharmaceutical composition in the
form of suppositories, a low-melting wax such as a
mixture of fatty acid glycerides and cocoa butter is
first melted and the active ingredient is dispersed
therein by, for example, stirring. The molten
homogeneous mixture is then poured into convenient-sized
molds and allowed to cool and solidify.
Powders and tablets preferably contain between
about 5o to about 70% by weight of the active ingredient.
Suitable carriers include, for example, magnesium
carbonate, magnesium stearate, talc, lactose, sugar,
pectin, dextrin, starch, tragacanth, methyl cellulose,
sodium carboxymethyl cellulose, a low-melting wax, cocoa
butter and the like.
The pharmaceutical compositions can include the
formulation of the active compound with encapsulating
material as a carrier providing a capsule in which the
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active component (with or without other carriers) is
surrounded by a carrier, which is thus in association
with it. In a similar manner, cachets are also included.
Tablets, powders, cachets, and capsules can be used as
5 solid dosage forms suitable for oral administration.
Liquid pharmaceutical compositions include, for
example, solutions suitable for oral or parenteral
administration, or suspensions, and emulsions suitable
for oral administration. Sterile water solutions of the
10 active component or sterile solutions of the active
component in solvents comprising water, ethanol, or
propylene glycol are examples of liquid compositions
suitable for parenteral administration.
Sterile solutions can be prepared by dissolving
15 the active component in the desired solvent system, and
then passing the resulting solution through a membrane
filter to sterilize it or, alternatively, by dissolving
the sterile compound in a previously sterilized solvent
under sterile conditions.
20 Aqueous solutions for oral administration can
be prepared by dissolving the active compound in water
and adding suitable flavorants, coloring agents,
stabilizers, and thickening agents as desired. Aqueous
suspensions for oral use can be made by dispersing the
25 finely divided active component in water together with a
viscous material such as natural or synthetic gums,
resins, methyl cellulose, sodium carboxymethyl cellulose,
and other suspending agents known to the pharmaceutical
formulation art.
30 Preferably, the pharmaceutical composition is
in unit dosage form. In such form, the composition is
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51
divided into unit doses containing appropriate quantities
of the active bicyclic hydantoin compound. The unit
dosage form can be a packaged preparation, the package
containing discrete quantities of the preparation, for
example, packeted tablets, capsules, and powders in vials
or ampules. The unit dosage form can also be a capsule,
cachet, or tablet itself, or it can be the appropriate
number of any of these packaged forms.
As pharmaceutical compositions for treating
infections, pain, or any other indication the compounds
of the present invention are generally in a
pharmaceutical composition so as to be administered.to a
subject at dosage levels of from 0.7 to 7000 mg per day,
and preferably 1 to 500 mg per day, for a normal human
adult of approximately 70 kg of body weight, this
translates into a dosage of from 0.01 to 100 mg/kg of
body weight per day. The specific dosages employed,
however, can be varied depending upon the requirements of
the patient, the severity of the condition being treated,
and the activity of the compound being employed. The
determination of optimum dosages for a particular
situation is within the skill of the art.
The compounds and combinatorial libraries of
the invention can be prepared as set forth in Figures 1
to 3 and as described below.
Variant bicyclic hydantoin derivative compounds
and combinatorial libraries can be prepared in order to
achieve a high level of diversity. For instance, as
shown in Figures 1 and 2, such compounds can be prepared
by coupling a molecule containing a group of the formula:
resin-NH- _C {O) -R,-NH-S {O~) -R~, wherein R1 and R."
independently, are each a variable group, with {b) a ring
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nitrogen compound that is substituted at the position
adjacent to the ring nitrogen with -C(0)-O-CH3, resulting
a ring nitrogen compound that is substituted at the
position adjacent to the ring nitrogen with -C(0)-O-CHI
and further substituted with a group of the formula
-N (S (Oz) -R.,) -Rz-C (O) -NH- (see step d of Figure 1 and step
d of Figure 2). The resulting compound can then react
with an isocyanate of the formula R3-NCO, where R3 is a
variable group, to form a -C(O)-NH-R~ group attached to
the ring nitrogen (see step f of Figure 1 and step f of
Figure 2). The resulting compound can then be cyclized
in the presence of a base (e.g., tetramethylguanidine) to
form a bicyclic hydantoin derivative (see step g of
Figure 1 and step g of Figure 2), which can then be
cleaved from the resin (see step h of Figure 1 and step h
of Figure 2).
The molecule containing a group of the formula
-NH-C (O) -R1-NH-S (O~) -Rz can be attached to a
functionalized resin, for example, MBHA, by reacting the
amino resin with protected (e. g., t-Boc) amino-R1-C(O)OH
(see step a of Figures 1 and 2) and then deprotecting
(see step b of Figures 1 and 2). The molecule containing
a group of the formula -NH-C (0) -R1-NH-S (O~) -Rz can be
formed by coupling a molecule containing a group of the
formula -NH-C (O) -R1-NHz with Rz-S (0a) -leaving group,
preferably, where the leaving group is a halide and, more
preferably, where the halide is chloride (see step c of
Figures 1 and 2).
The the ring nitrogen compound can be
five-member (e.g., a pyrrolidine derivative; see step d
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53
of Figure l) and, therefore, the resulting compound can
be of the formula:
O
o ~\
NF~3
HN- ~' II
0
(see steps a to h of Figure 1).
However, the ring nitrogen compound can also be
a piperidine derivative (e.g, a six-member ring; see
steps c and d of Figure 2) and can also be a seven-member
ring or even an eight-member ring. It should be
understood that, where the ring is greater than five-
member, it can contain a double bond. It should also be
understood that the ring can be fused to another ring,
such as a phenyl ring.
In addition, a ring nitrogen compound with a
group containing -C(O)-0- attached (and directly attached
to the carbonyl carbon) to a position adjacent to the
ring nitrogen (see steps a and b of Figure 3) can be
coupled with an isocyanate derivative of the formula
variable-NCO to form a ring nitrogen compound with the
group -C(O)-NH-variable directly attached to the ring
nitrogen (see step c of Figure 3). The resulting
compound can be cyclized in the presence of a base, for
example, tetramethylguanidine or barium hydroxide, to
form a bicyclic hydantoin derivative (see step c of
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Figure 3). The group containing -C(O)-0- can be attached
to a functionalized resin, for example, via oxygen (see
step a of Figure 3). The ring nitrogen can be five- six-
or seven or even eight-member (see Figure 3).
Resin-bound bicyclic hydantoin derivative
compounds can be cleaved by treating them, for example,
with HF. They can also be cleaved with TFA/DCM, provided
that TFA sensitive protecting group such as Boc are not
used in the synthetic scheme. The compounds can be
extracted from the spent resin, for example, with AcOH.
Bicyclic hydantoin derivative compounds and
libraries, such~as~those of the present invention, can be
made utilizing individual polyethylene bags, referred to
as "tea bags" (see Houghten et al., Proc. Natl. Acad.
Sci. USA 82: 5131 (1985); Biochemistry, 32:11035 (1993);
and U.S. Patent No. 4,631,211, all of which are
incorporated herein by reference).
The nonsupport-bound combinatorial libraries
can be screened as single compounds. In addition, the
nonsupport-bound combinatorial libraries can be screened
as mixtures in solution in assays such as radio-receptor
inhibition assays, anti-bacterial assays, anti-fungal
assays, calmodulin-dependent phosphodiesterase (CaMPDE)
assays and phosphodiesterase (PDE) assays, as described
in detail below. Deconvolution of highly active mixtures
can then be carried out by iterative or positional
scanning methods. These techniques, the iterative
approach or the positional scanning approach, can be
utilized for finding other active compounds within the
combinatorial libraries of the present invention using
any one of the below-described assays or others well
known in the art.
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The iterative approach is well-known and is set
forth in general in Houghten et al., Nature, 354, 84-86
(1991) and Dooley et al., Science, 266, 2019-2022 (1994),
both of which are incorporated herein by reference. In
5 the iterative approach, for example, sub-libraries of a
molecule having three variable groups are made wherein
the first variable is defined. Each of the compounds
with the defined variable group is reacted with all of
the other possibilities at the other two variable groups.
10 These sub-libraries are each tested to define the
identity of the second variable in the sub-library having
the highest activity in the screen of choice. A new sub-
library with the first two variable positions defined is
reacted again with all the other possibilities at the
15 remaining undefined variable position. As before, the
identity of the third variable position in the sub-
library having the highest activity is determined. If
more variables exist, this process is repeated for all
variables, yielding the compound with each variable
20 contributing to the highest desired activity in the
screening process. Promising compounds from this process
can then be synthesized on larger scale in traditional
single-compound synthetic methods for further biological
investigation.
25 The positional-scanning approach has been
described for various combinatorial libraries as
described, for example, in R. Houghten et al.
PCT/US91/08694 and U.S. Patent 5,556,762, both of which
are incorporated herein by reference. In the positional
30 scanning approach, sublibraries are made defining only
one variable with each set of sublibraries and all
possible sublibraries with each single variable defined
(and all other possibilities at all of the other variable
positions), made and tested. From the instant
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description one skilled in the art could synthesize
combinatorial libraries wherein two fixed positions are
defined at a time. From the testing of each single-
variable defined combinatorial library, the optimum
substituent at that position can be determined, pointing
to the optimum or at least a series of compounds having a
maximum of the desired biological activity. Thus, the
number of sublibraries for compounds with a single
position defined will be the number of different
substituents desired at that position, and the number of
all the compounds in each sublibrary will be the product
of the number of substituents at each of the other
variables.
Individual compounds and pharmaceutical
compositions containing the compounds, as well as methods
of using the same, are included within the scope of the
present invention. The compounds of the present
invention can be used for a variety of purposes and
indications and as medicaments for any such purposes and
indications. For example, bicyclic hydantoin derivative
compounds of the present invention can be used as
pesticides, acaricides, receptor agonists or antagonists
and antimicrobial agents, including antibacterial or
antiviral agents. The libraries can be screened in any
variety of melanocortin receptor and related activity
assays, such as those detailed below as well as others
known in the art. Additionally, the subject compounds
can be useful as analgesics. Assays which can be used to
test the biological activity of the instant compounds
include antimicrobial assays, a competitive enzyme-linked
immunoabsorbent assay and radio-receptor assays, as
described below.
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The melanocortin (MC) receptors are a group of
cell surface proteins that mediate a variety of
physiological effects, including regulation of adrenal
gland function such as production of the glucocorticoids
cortisol and aldosterone; control of melanocyte growth
and pigment production; thermoregulation;
immunomodulation; and analgesia. Five distinct
MC receptors have been cloned and are expressed in a
variety of tissues, including melanocytes, adrenal
cortex, brain, gut, placenta, skeletal muscle, lung,
spleen, thymus, bone marrow, pituitary,' gonads and
adipose tissue (Tatro, Neuroimmunomodulation 3:259-284
(1996)). Three MC receptors, MCR-1, MCR-3 and MCR-4, are
expressed in brain tissue (Xia et al., Neuroreport
6:2193-2196 (1995)).
A variety of ligands termed melanocortins
function as agonists that stimulate the activity of
MC receptors. The melanocortins include
melanocyte-stimulating hormones (MSH) such as a-MSH,
(3-MSH and y-MSH, as well as adrenocorticotropic hormone
(ACTH). Individual ligands can bind to multiple
MC receptors with differing relative affinities. The
variety of ligands and MC receptors with differential
tissue-specific expression likely provides the molecular
basis for the diverse physiological effects of
melanocortins and MC receptors. For example, a-MSH
antagonizes the actions of immunological substances such
as cytokines and acts to modulate fever, inflammation and
immune responses (Catania and Lipton, Annals N. Y. Acad.
Sci. 680:412-423 (1993)).
The role of certain specific MC receptors in
some of the physiological effects described above for MC
receptors has been elucidated. For example, MCR-1 is
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involved in pain and inflammation. MCR-1 mRNA is
expressed in neutrophils (Catania et al., Peptides
17:675-679 (1996)). The anti-inflammatory agent a-MSH
was found to inhibit migration of neutrophils. Thus, the
presence of MCR-1 in neutrophils correlates with the
anti-inflammatory activity of a-MSH.
An interesting link of MC receptors to
regulation of food intake and obesity has recently been
described. The brain MC receptor MCR-4 has been shown to
function in the regulation of body weight and food
intake. Mice in which MCR-4 has been knocked out exhibit
weight gain (Huszar et al., Cell 88:131-141 (1997)). In
addition, injection into brain of synthetic peptides that
mimic melanocortins and bind to MCR-4 caused suppressed
feeding in normal and mutant obese mice (Fan et al.,
Nature 385:165-168 (1997)). These results indicate that
the brain MC receptor MCR-4 functions in regulating food
intake and body weight.
Due to the varied physiological activities of
MC receptors, high affinity ligands of MC receptors could
be used to exploit the varied physiological responses of
MC receptors by functioning as potential therapeutic
agents ar as lead compounds for the development of
therapeutic agents. Furthermore, due to the effect of MC
receptors on the activity of various cytokines, high
affinity MC receptor ligands could also be used to
regulate cytokine activity.
A variety of assays can be used to identify or
characterize MC receptor ligands of the invention. For
example, the ability of a bicyclic hydantoin derivative
compound to compete far binding of a known MC receptor
ligand can be used to assess the affinity and specificity
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of a bicyclic hydantoin derivative compound for one or
more MC receptors. Any MC receptor ligand can be used so
long as the ligand can be labeled with a detectable
moiety. The detectable moiety can be, for example, a
radiolabel, fluorescent label or chromophore, or any
detectable functional moiety so long as the MC receptor
ligand exhibits specific MC receptor binding. A
particularly useful detectable MC receptor ligand for
identifying and characterizing other MC receptor ligands
is '-SST-HP 467, which has the amino acid sequence
Ac-Nle-Gln-His- (p (I ) -D-Phe) -Arg- (D-Trp) -Gly-NH_ and is
described in Dooley et al., "Melanocortin Receptor
Ligands and Methods of Using Same," U.S. patent
application 091027,108, filed February 20, 1998, which is
incorporated herein by reference. HP 467 is a para-
iodinated form of HP 228.
Using assay methods such as those described
above, binding kinetics and competition with radiolabeled
HP 467 can confirm that bicyclic hydantoin derivative
compounds of the invention bind to one or more MC
receptors. Furthermore, bicyclic hydantoin derivative
compounds of the invention can exhibit a range of
affinities and specificity for various MC receptors.
The invention provides MC receptor ligands that
can bind to several MC receptors with similar affinity.
In addition, the invention also provides MC receptor
ligands that can be selective for one or more MC
receptors. As used herein, the term "selective" means
that the affinity of a MC receptor ligand differs between
one MC receptor and another by about 10-fold, generally
about 20- to 50-fold, and particularly about 100-fold.
In some cases, a MC receptor ligand having broad
specificity is desired. In other cases, it is desirable
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to use MC receptor ligands having selectivity for a
particular MC receptor. For example, MCR-1 ligands are
particularly useful for treating pain and inflammation,
whereas MCR-4~ligands are useful for treating obesity.
5 The binding characteristics and specificity of a given MC
receptor ligand can be selected based on the particular
disease or physiological effect that is desired to be
altered.
Another assay useful for identifying or
10 characterizing MC receptor ligands measures signaling of
MC receptors. MC receptors are G protein-coupled
receptors that couple to adenylate cyclase and produce
cAMP. Therefore, measuring cAMP production in a cell
expressing a MC receptor and treated with a MC receptor
15 ligand can be used to assess the function of the MC
receptor ligand in activating a MC receptor.
Ligands for MC-3 that can alter the activity of
an MC-3 receptor can be useful for treating sexual
dysfunction and other conditions or conditions associated
20 with MC-3 such as inflammation. Other MC-3-associated
conditions that can be treated with the MC-3 receptor
ligands include disuse deconditioning; organ damage such
as organ transplantation or ischemic injury; adverse
reactions associated with cancer chemotherapy; diseases
25 such as atherosclerosis that are mediated by free
radicals and nitric oxide action; bacterial endotoxic
sepsis and related shock; adult respiratory distress
syndrome; and autoimmune or other patho-immunogenic
diseases or reactions such as allergic reactions or
30 anaphylaxis, rheumatoid arthritis, inflammatory bowel
disease, ulcerative colitis, glomerulonephritis, systemic
lupus erythematosus, transplant atherosclerosis and
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parasitic mediated immune dysfunctions such as Chagas's
disease.
The invention further provides a method for
treating an MC-3-associated condition in a subject. The
term "MC-3-associated condition" includes any condition
or condition mediated by MC-3 or can be affected by
binding an MC-3 ligand. Such conditions include
inflammation and sexual dysfunction.
The term "sexual dysfunction" herein means any
condition that inhibits or impairs normal sexual
function, including coitus. However, the term need not
be limited to physiological conditions, but may include
psychogenic conditions or perceived impairment without a
formal diagnosis of pathology.
In males, sexual dysfunction includes erectile
dysfunction. The term "erectile dysfunction" or
"impotence" means herein the inability or impaired
ability to attain or sustain an erection that would be of
satisfactory rigidity for coitus. Sexual dysfunction in
males can also include premature ejaculation and
priapism, which is a condition of prolonged and sometimes
painful erection unrelated to sexual activity, often
associated with sickle-cell disease.
In females, sexual dysfunction includes sexual
arousal disorder. The term "sexual arousal disorder"
means herein a persistent or recurrent failure to attain
or maintain the lubrication-swelling response of sexual
excitement until completion of sexual activity. Sexual
dysfunction in females can also include inhibited orgasm
and dyspareunia, which is painful or difficult coitus.
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Sexual dysfunction can also be manifested as inhibited
sexual desire or inhibited lordosis behavior in animals.
In addition, the ability of the compounds to
inhibit bacterial growth, and therefore be useful to that
infection, can be determined by methods well known in the
art. Compounds of the present invention can be shown to
have antimicrobial activity by the in vitro antimicrobial
activity assay described below and, therefore, are useful
as antimicrobial agents.
Moreover, an exemplary in vitro antimicrobial
activity assay is described in Blondelle and Houghten,
Biochemistry 30:4671-4678 (1991), which is incorporated
herein by reference. In brief, Staphylococcus aureus
ATCC 29213 (Rockville, MD) is grown overnight at 37°C in
Mueller-Hinton broth, then re-inoculated and incubated at
37°C to reach the exponential phase of bacterial growth
(i.e., a final bacterial suspension containing 105 to 5 x
105 colony-forming units/ml). The concentration of cells
is established by plating 100 p1 of the culture solution
using serial dilutions (e. g. , 10--', 10-3 and 10-4) onto
solid agar plates. In 96-well tissue culture plates,
compounds, individual or in mixtures, are added to the
bacterial suspension at concentrations derived from
serial two-fold dilutions ranging from 1500 to 2.9 pg/ml.
The plates are incubated overnight at 37°C and the growth
determined at each concentration by OD62o nm. The ICSo
(the concentration necessary to inhibit 50% of the growth
of the bacteria) can then be calculated.
The competitive ELISA method which can be used
here is a modification of the direct ELISA technique
described previously in Appel et al., J. Immunol.
144:976-983 (1990), which is incorporated herein by
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63
reference. It differs only in the MAb addition step.
Briefly, mufti-well microplates are coated with the
antigenic peptide (Ac-GASPYPNLSNQQT-NH,) at a
concentration of 100 pmol/50 ul. After blocking, 25.u1
of a 1.0 mg/ml solution of each mixture of a synthetic
combinatorial library (or individual compound) is added,
followed by MAb 125-10F3 (Appel et al., supra) (25 pl per
well). The MAb is added at a fixed dilution in which the
bicyclic guanidine in solution effectively competes for
MAb binding with the antigenic peptide adsorbed to the
plate. The remaining steps are the same as for direct
ELISA. The concentration of compound necessary to
inhibit 50% of the MAb binding to the control peptide on
the plate (IC,o) is determined by serial dilutions of the
compound.
Alternative screening can be done with radio-
receptor assays. The radio-receptor assay, can be
selective for any one of the u, K, or c5 opiate receptors.
Compounds.of the present invention can be useful in vitro
for the diagnosis of relevant opioid receptor subtypes,
such as K, in the brain and other tissue samples.
Similarly, the compounds can be used in vivo
diagnostically to localize opioid receptor subtypes.
The radio-receptor assays are also an
indication of the compounds' analgesic properties as
described, for example, in Dooley et al., Proc. Natl.
Acad. Sci., 90:10811-10815 (1993). For example, it can
be envisioned that these compounds can be used for
therapeutic purposes to block the peripheral effects of a
centrally acting pain killer. For instance, morphine is
a centrally acting pain killer. Morphine, however, has a
' numlaer of deleterious effects in the periphery which are
not required for the desired analgesic effects, such as
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64
constipation and pruritus (itching). While it is known
that the many compounds do not readily cross the blood-
brain barrier and, therefore, elicit no central effect,
the subject compounds can have value in blocking the
periphery effects of morphine, such as constipation and
pruritus. Accordingly, the subject compounds can also be
useful as drugs, namely as analgesics, or to treat
pathologies associated with other compounds which
interact with the opioid receptor system.
Additionally, such compounds can be tested in a
o receptor assay. Ligands for the 6 receptor can be
useful as antipsychotic agents, as described in Abou-
Gharbia et al., Annual Reports in Medicinal Chemistry,
28:1-10 (1993).
Radio-receptor assays can be performed with
particulate membranes prepared using a modification of
the method described in Pasternak et al., Mol. Pharmacol.
11:340-351 (1975), which is incorporated herein by
reference. Rat brains frozen in liquid nitrogen can be
obtained from Rockland (Gilbertsville, PA). The brains
are thawed, the cerebella removed and the remaining
tissue weighed. Each brain is individually homogenized
in 40 ml Tris-HCl buffer (50 mM, pH 7.4, 4°C) and
centrifuged (Sorvall° RCSC SA-600: Du Pont, Wilmington,
DE) (16,000 rpm) for 10 minutes. The pellets are
resuspended in fresh Tris-HC1 buffer and incubated at
37°C for 40 minutes. Following incubation, the
suspensions are centrifuged as before, the resulting
pellets resuspended in 100 volumes of Tris buffer and the
suspensions combined. Membrane suspensions are prepared
and used in the same day. Protein content of the crude
homogenates generally range from 0.15-0.2 mg/ml as
determined using the method described in Bradford, M.M.,
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Anal. Biochem. 72:248-254 (1976), which is incorporated
herein by reference.
Binding assays are carried out in polypropylene
tubes, each tube containing 0.5 ml of membrane
5 suspension. 8 nM of 3H- [ D-Alaz, Me-Pheq, Gly-ol'] enkephalin
(DAMGO) (specific activity = 36 Ci/mmol, 160,000 cpm per
tube; which can be obtained from Multiple Peptide
Systems, San Diego, CA, through NIDA drug distribution
program 271-90-7302) and 80 ug/ml of bicyclic guanidine,
10 individual or as a mixture and Tris-HCl buffer in a total
volume of 0.65 ml. Assay tubes are incubated for 60
mins. at 25°C. The reaction is terminated by filtration
through GF-B filters on a Tomtec harvester (Orange, CT).
The filters are subsequently washed with 6 ml of Tris-HCl
15 buffer, 4°C. Bound radioactivity is counted on a
Pharmacia Biotech Betaplate Liquid Scintillation Counter
(Piscataway, NJ) and expressed in cpm. To determine
inter- and intra-assay variation, standard curves in
which 3H-DAMGO is incubated in the presence of a range of
20 concentrations of unlabeled DAMGO (0.13-3900 nM) are
generally included in each plate of each assay (a 96-well
format). Competitive inhibition assays are performed as
above using serial dilutions of the bicyclic guanidines,
individually or in mixtures. ICSO values (the
25 concentration necessary to inhibit 500 of 3H-DAMGO
binding) are then calculated. ICSO values of less than
1000 nM are indicative of highly active opioid compounds
which bind to the a receptor, with particularly active
compounds having ICso values of 100 nM or less and the
30 most active compounds with values of less than 10 nM.
As opposed to this a receptor selective assay,
which can be carried out using 'H-DAMGO as radioligand, as
described above, assays selective for K receptors can be
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carried out using [~H]-U69,593 (3 nM, specific activity 62
Ci/mmol) as radioligand. Assays selective for b opiate
receptors can be carried out using tritiated DSLET ([D-
Ser'-, D-Leus]-threonine-enkephalin) as radioligand.
Assays selective for the 6 opiate receptor can use
radiolabeled pentazocine as ligand.
Screening of combinatorial libraries and
compounds of the invention can be done with an
anti-fungal assay. Compounds of the present invention
can be useful for treating fungal infections.
Screening of combinatorial libraries and
compounds of the invention also can be done with a
calmodulin-dependent phosphodiesterase (CaMPDE) assay.
Compounds of the present invention can be useful as
calmodulin antagonists.
Calmodulin (CaM), which is the major
intracellular calcium receptor, is involved in many
processes that are crucial to cellular viability. In
particular, ealmodulin is implicated in calcium-
stimulated cell proliferation. Calmodulin antagonists
are, therefore, useful for treating conditions associated
with increased cell proliferation, for example, cancer.
In addition, calmodulin antagonists such as compounds of
the subject invention are useful both in vitro and in
vivo for identifying the role of calmodulin in other
biological processes. The disadvantages of )mown
antagonists such as trifluoperazine and N-(4-aminobutyl)-
5-chloro-2-naphthalenesulfonamide (W13) include their
non-specificity and toxicity. In contrast, advantages of
the combinatorial libraries and compounds of the subject
invention as calmodulin antagonists include their reduced
flexibility and ability to generate broader
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conformational space of interactive residues as compared
to their linear counterparts.
An example of an assay that identifies CaM
antagonists is a CaMPDE assay. In brief, samples are
mixed with 50 ~l of assay buffer (360 mM Tris, 360 mM
Imidazole, 45 mM Mg(CH3C00)" pH 7.5) and 10 ~1 of CaCla
(4.5 mM) to a final volume of 251 ~l. 25 u1 of
calmodulin stock solution (Boehringer Mannheim;
0.01 ug/ul) is then added and the samples then sit at
room temperature for 10 minutes. 14 u1 of PDE (Sigma; 2
Units dissolved in 4 ml of water; stock concentration:
0.0005 Units/ul) is then added, followed by 50 u1 of
5'-nucleotidase (Sigma; 100 Units dissolved in 10 ml of
10 mM Tris-HCl containing 0.5 mM Mg(CH3C00)_, pH 7.0;
stock concentration: 10 Units/ml). The samples are then
incubated for 10 minutes at 30°C. 50 u1 of adenosine
3',5'-cyclic monophosphate (CAMP) (20 mM in water at pH
7.0) is added, the samples incubated for 1 hour at 30°C
and then vortexed. 200 u1 of trichloroacetic acid (TCA)
(55o in water) is added to a 200 u1 sample aliquot, which
is then vortexed and centrifuged for 10 minutes. 80 ~1
of the resulting supernatants of each sample is
transferred to a 96-well plate, with 2 wells each
containing 80 u1 of each sample. 80 u1 of ammonium'
molybdate (1.1o in 1.1N HzS04) is then added to all the
wells, and the OD of each were determined at 730nm, with
the values later subtracted to the final OD reading.
16 u1 of reducing agent (6g sodium bisulfate, 0.6g sodium
sulfite and 125mg of 1-amino-2-naphtol-4-sulfonic acid in
50m1 of water) is then added to one of each sample
duplicate and 16 u1 of water is added to the other
duplicate. After sitting for 1 hour at room temperature,
the OD of each well is determined at 730nm. The percent
inhibition of calmodulin activity is then calculated for
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each sample, using as Oo inhibition a control sample
containing all reagents without any test samples and as
1000 inhibition a control sample containing test samples
and all reagents except calmodulin. In addition, the
percent inhibition of phosphodiesterase activity was
determined by following a similar protocol as the CaMPDE
assay described above, except not adding calmodulin to
the sample mixture and calculating the percent inhibition
by using as Oo inhibition a control reagent without any
test samples and as 1000 inhibition a control sample
containing test samples and all reagents except CAMP.
The following examples are provided to
illustrate but not~limit the present invention. The
following abreviations have the corresponding meanings:
MBHA . 4-methylbenzhydrylamine;
DMF . N,N-dimethylforamide;
HOBt . l-hydroxybenzotriazole;
DMSO . dimethylsulfoxide;
Boc , tert-butoxycarbonyl;
FMOC . 9-fluorenyl-methoxycarbonyl;
DMAP . 4-dimethylamino-pyridine;
DIC . N,N'-diisopropylcarbodiimide;
TFA . trifluoroacetic acid;
DIEA . N,N-diisopropylethylamine;
DCM . dichloromethane;
TMOF: trimethylorthoformate;
HATU . azabenzotriazolyl-N,N,N',N'-tetramethyluronium
hexafluorophosphate;
CDI . carbonyldiimidazole
NMP . N-methylpyrrolidinone
DMA . N,N-dimethyl acetamide
RT . room temperature
IPA . isopropyl alcohol
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MeOH: methanol
MeOEtOH . 2-methoxyethanol
DCE . 1,2-dichloroethane
THF . tetrahydrofuran
ACN . acetonitrile
Wang resin . p-benzyloxybenzyl alcohol-polystyrene
Br-Wang resin . p-benzyloxybenzyl bromide-polystyrene
PP . polypropylene
PPh3Br2 . triphenylphosphine dibromide
DMAP . 4-dimethylamino-pyridine
KOtBu . potassium tert-butoxide
NaOMe . sodium methoxide
BtCH2CN . 1-(cyanomethyl)benzotriazole
DBU . 1,8-diazabicyclo[5.4.0]under-7-ene
Boc . tertbutoxycarbonyl;
AcOH . acetic acid
HPLC/MS . high performance liquid chromatography - mass
spectrometry;
FIA-MS . flow injection analysis - mass spectrometry
ELSD . evaporative light scattering detector
THB . Todd Hewitt Broth
OD , optical density
Example 1
Preparation of Bicyclic Hydantoins
Step a: Coupling of N-Boc-amino acid to MBHA and Boc
deprotection.
MBHA-HCl resin (methylbenzhydrylamine hydrochloride)
(1.6 g, 2.34 mmol NHz) was dispensed into a porous
polypropylene packet (Tea-bag, 75mm x 80mm, 65~). The
resin in the packet was neutralized with 5o DIEA/DCM
(3X80 mL) and then washed with DCM (80 mL). The packet
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was put in a Nalgene bottle and DMF (78 mL), Boc-L-
phenylalanine (6.18 g, 23.3 mmol), DIEA (3.01 g, 23.3
mmol ) , HOBT ( 2 . 62 g, 19 . 4 mmol ) and DIC ( 2 . 94 g, 23 . 3
mmol) were added sequentially. After shaking the bottle
5 for 20 hours, the packet was washed with DMF (3X80 mL),
DCM (3X80 mL) and MeOH (3X80 mL) and dried in air for
overnight. The packet was shaken with 55o TFA/DCM (80
mL) at room temperature for 30 minutes. It was then
washed with DCM (3X80 mL), 5o DIEA/DCM (3X80 mL) and MeOH
10 (2X80 mL) and dried in air for overnight.
The following protected amino acids were coupled to
resin according to the above-described procedures:
BOC-L-PHENYLALANINE
BOC-L-ALANINE
15 BOC-L-2-AMINOBUTYRIC ACID
BOC-4-AMINOBUTYRIC ACID
BOC-6-AMINOHEXANOIC ACID
BOC-4-CHLORO-L-PHENYLALANINE
BOC-L-METHIONINE
20 BOC-O-METHYL-L-TYROSINE
BOC-L-VALINE
Na-BOC-Nlm-TOSYL-L-HI ST I DINE
BOC-0-BENZYL-L-SERINE
BOC-L-LEUCINE
25 BOC-3-CYCLOHEXYL-L-ALANINE
4-(BOC-AMINOMETHYL)BENZOIC ACID
4-(BOC-AMINOMETHYL)CYCLOHEXANECARBOXYLIC ACID
Step b: Sulfonylation of amine with sulfonyl chloride.
30 The packet from step a was shaken with a solution of
DIEA (2.89 g, 22.4 mmol), 2-thiophene sulfonyl chloride
(3.41 g, 18.69 mmol) in DCM (75 mL) for 48 hours until
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ninhydrin Kaiser test of a few resin beads taken from the
packet was negative. The packet was washed thoroughly
with DCM (3X80 mL), DMF (3X80 mL) and t-butylmethyl ether
(3X80 mL), then dried in air overnight. The following
sulfonyl chlorides were also added according to the
procedure of step b:
2-THIOPHENESULFONYL CHLORIDE
BENZENESULFONYL CHLORIDE
2,5-DICHLOROBENZENESULFONYL CHLORIDE
2-NITROBENZENESULFONYL CHLORIDE
4-BROMOBENZENESULFONYL CHLORIDE
4-FLUOROBENZENESULFONYL CHLORIDE
4-CHLOROBENZENESULFONYL CHLORIDE
3-(TRIFLUOROMETHYL)BENZENESULFONYL CHLORIDE
3,4-DICHLOROBENZENESULFONYL CHLORIDE
3-CHLORO-4-FLUOROBENZENESULFONYL CHLORIDE
2-FLUOROBENZENESULFONYL CHLORIDE
3-FLUOROBENZENESULFONYL CHLORIDE
4-(TRIFLUOROMETHYL)BENZENESULFONYL CHLORIDE
2,4-DIFLUOROBENZENESULFONYL CHLORIDE
2-CHLOROBENZENESULFONYL CHLORIDE
2-(TRIFLUOROMETHYL)BENZENESULFONYL CHLORTDE
3-CHLOROBENZENESULFONYL CHLORIDE
3,5-DICHLOROBENZENESULFONYL CHLORIDE
2,3-DICHLOROBENZENESULFONYL CHLORIDE
2-BROMOBENZENESULFONYL CHLORIDE
5-(2-PYRIDYL)THIOPHENE-2-SULFONYL CHLORIDE
2-CHLORO-5-(TRIFLUOROMETHYL)BENZENESULFONYL CHLORIDE
4-CYANflBENZENESULFONYL CHLORIDE
2-CYANOBENZENESULFONYL CHLORIDE
5-CHLORO-1,3-DIMETHYLPYRAZOLE-4-SULPHONYL CHLORIDE
3,5-DIMETHYLISOXAZOLE-4-SULFONYL CHLORIDE
2,4-DICHLOROBENZENESULFONYL CHLORIDE
2-CHLORO-4-(TRIFLUOROMETHYLBENZENE)SULFONYL CHLORIDE
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2-CHLORO-4-FLUOROBENZENESULFONYL CHLORIDE
2,4,6-TRICHLOROBENZENESULFONYL CHLORIDE
1-METHYLIMIDAZOLE-4-SULPHONYL CHLORIDE
METHYL 3-CHLOROSULFONYLTHIOPHENE-2-CARBOXYLATE
5-ISOXAZOL-3-YLTHIOPHENE-2-SULFONYL CHLORIDE
4-BIPHENYLSULFONYL CHLORIDE
3,4-DIFLUOROBENZENESULPHONYL CHLORIDE
3-METHYL-5-CHLORO-2-BENZOTHIOPHENE
3-CYANOBENZENESULPHONYL CHLORTDE
4-METHYLSULFONYLBENZENESULFONYL CHLORIDE
2-METHYLSULFONYLBENZENESULFONYL CHLORIDE
Step c: Formation of 4-aminoproline derivative on resin
via the Mitsunobu reaction and Boc deprotection.
The packet from step b was shaken in a glass bottle
with a 0.35,M solution of trans-L-4-hydroxyproline methyl
ester (4.58 g, 18.69 mmol), triphenylphosphine (4.90 g,
18.69 mmol), NMM (N-methylmorpholine) (17.8 mL), THF
(17.8 mL) and DCM (17.8 mL) for 5 minutes. The mixture
was then cooled to 0-5°C for 2 hours in a refrigerator.
The glass bottle was then equipped with a thermometer, an
additional funnel, a nitrogen stream inlet, an ice water
bath and an electric shaker. A solution of DIAD
(diisopropyl azodicarboxylate) (3.78 g, 18.69 mmol) in
DCM (5 mL) was added dropwise under nitrogen atmosphere
while the bottle was shaken. The reaction mixture was
maintained at 0-5°C with an ice water bath during the
addition of DIAD. The reaction mixture was then shaken
at room temperature for 3 days. The packet was then
washed thoroughly with DMF (3X80 mL), DCM (3X80 mL) and
MeOH (3X80 mL) and dried in air overnight. The packet
was shaken with 55o TFA/DCM (80 mL) at room temperature
for 30 minutes and washed with DCM (3X80 mL), 5o DIEA/DCM
(3X80 mL) and MeOH (3X80 mL) and dried in air overnight.
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Step d: Reaction of isocyanate to form urea.
The resin~was removed from the packet and suspended
in DMF (20 mL) and the resin suspension was distributed
equally into 40 wells of a microtiter plate (2 mL X 40).
The extra DMF was removed from the wells, leaving about
0.2 mL of DMF in each well. An array of 40 different
isocyanate solutions (0.5 M in DMF) was prepared. To
each well was added 1.0 mL of the following isocyanate in
solution:
PHENYL ISOCYANATE
2-BROMOPHENYL ISOCYANATE
2-FLUOROPHENYL ISOCYANATE
2,4-DIFLUOROPHENYL TSOCYANATE
2,6-DIFLUOROPHENYL ISOCYANATE
2-CHLOROPHENYL ISOCYANATE
2,4-DICHLOROPHENYL ISOCYANATE
2,5-DICHLOROPHENYL ISOCYANATE
2-METHOXYPHENYL ISOCYANATE
O-TOLYL ISOCYANATE
2-ETHYLPHENYL ISOCYANATE
3-FLUOROPHENYL ISOCYANATE
3-CHLOROPHENYL ISOCYANATE
3-METHQXYPHENYL ISOCYANATE
M-TOLYL ISOCYANATE
4-BROMOPHENYL ISOCYANATE
4-FLUOROPHENYL ISOCYANATE
4-METHOXYPHENYL ISOCYANATE
P-TOLYL ISOCYANATE
1-NAPHTHYL ISOCYANATE
BENZYL ISOCYANATE
2-ISOPROPYLPHENYL ISOCYANATE
2,9-DIMETHYLPHENYL ISOCYANATE
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2,5-DIMETHYLPHENYL ISOCYANATE
2-ETHYL-6-METHYLPHENYL ISOCYANATE
3-(METHYLTHIO)PHENYL ISOCYANATE
3,4-DIMETHYLPI-IENYL ISOCYANATE
3,5-DIMETHYLPHENYL ISOCYANATE
2-METHOXY-5-METHYLPHENYL ISOCYANATE
3-ETHYLPHENYL ISOCYANATE
4-ETHOXYPHENYL ISOCYANATE
4-(METHYLTHIO)PHENYL ISOCYANATE
4-ISOPROPYLPHENYL ISOCYANATE
4-ETHYLPHENYL ISOCYANATE
4-N-BUTYLPHENYL ISOCYANATE
2-ISOPROPYL-6-METHYLPHENYL ISOCYANATE
2,4,5-TRIMETHYLPHENYL ISOCYANATE
4-BUTOXYPHENYL ISOCYANATE
5-FLUORO-2-METHYLPHENYL ISOCYANATE
4-(DTMETHYLAMTNO)PHENYL ISOCYANATE
The plate was capped tightly and shaken at room
temperature for 3 d. The resin was washed with DMF
(3XlmL/well), MeOH (3XlmL/well) and DMF(3XlmL/well). The
extra DMF was removed, leaving about 0.2 mL of DMF in
each well.
Step e: Tetramethylguanidine-mediated cyclization to
bicyclic hydantoin.
To each well of the microtiter plate was added
1.1 mL of 0.025 M TMG (tetramethyl guanidine). The plate
was capped and shaken at room temperature for 16 h. The
resin was washed with DMF (3XlmL/well), MeOH
(3XlmL/well), DMF (3XlmL/well), MeOH (6XlmL/well). The
resin was dried in air for three days and under vacuum
for overnight. The plate was treated with gaseous HF at
room temperature for 2 hours. The HF was removed
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completely by a stream of nitrogen and then by Vacuum.
The resin was extracted with acetic acid (3X0.5 mL/well).
The extract was lyophilized to give the title compounds.
A library of about 8,000 individual bicyclic
5 hydantoins was synthesized using 15 amino acids (R1), 39
sulfonyl chlorides (R2) and 40 isocyanates (R3), as
listed above. All compounds were analyzed using FIA-MS.
Example 2
Anti-microbial Screen
10 Streptococcus pyogenes (ATCC# 97-03 14289) was
grown in Todd Hewitt Broth (THB) (Difco Laboratories
#0492-17-6) overnight, until reaching an optical density
of ( OD = 0.636@ 570 nm) by reading 0.1 ml in a 96 well
microtiter plate in a Molecular Devices Thermomax. This
15 preparation was kept frozen as stocks in 30o v/v glycerol
in 1.5 ml aliquots at -70mC until use. Prior to
experiments, 6 ml aliquots were thawed and diluted into
50 ml 2X THB. 60 u1 of this dilution was added to 92
wells of microtiter plate. To three wells THB (200 u1)
20 was added to serve as a blank and a sterility control.
Test compounds in DMSO and appropriate concentrations of
DMSO were added to Growth/Solvent Controls at 0 time.
Plates were read at 0 time at 570 nm in the Molecular
Devices plate reader to obtain compounds correction
25 factors for insoluble or colored compounds. Plates were
read again at 4 hours.
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Percent inhibition is calculated with the following
formulae:
Color correct =
(0.D. 0 hr - Blank 0 hr)-(Solvent Control Ohr - Blank 0
hr)
o Inhibition =
100 - O.D. test compound 4hr -Blank 4 hr - color
correct
O.D. growth/solvent control 4 hr - Blank 4 hr
Under this formula, the most active compounds
tested were as follows:
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77
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CA 02416654 2003-O1-20
WO 02/08227 PCT/EPO1/08322
78
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CA 02416654 2003-O1-20
WO 02/08227 PCT/EPO1/08322
79
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CA 02416654 2003-O1-20
WO 02/08227 PCT/EPO1/08322
a
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CA 02416654 2003-O1-20
WO 02/08227 PCT/EPO1/08322
81
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EXAMPLE 3
Melanocartin Receptor Assay
This example describes methods for assaying
binding to MC receptors.
All cell culture media and reagents are
obtained from GibcoBRL (Gaithersburg MD), except for
COSMIC CALF SERUM (HyClone; Logan UT). HEK 293 cell
lines are transfected with the human MC receptors hMCR-1,
hMCR-3, and hMCR-4 (Gantz et al., Biochem. Biophys. Res.
Comm. 200:1214-1220 (1994); Gantz et al., J. Biol. Chem.
268:8246-8250 (1993); Gantz et al. J. Biol. Chem.
268:15174-15179 (1993); Haskell-Leuvano et al., Biochem.
Biophys. Res. Comm. 204:1137-1142 (1994); each of which
is incorporated herein by reference). Vectors for
construction of an hMCR-5 expressing cell line are
obtained, and a line of HEK 293 cells expressing hMCR-5
is constructed (Gantz, supra, 1994). hMCR-5 has been
described previously (Franberg et al., Biochem. Biophys.
Res. Commun. 236:489-492 (1997); Chowdhary et al.,
Cytoaenet. Cell Genet. 68:1-2 (1995); Chowdhary et al.,
Cytoaenet. Cell Genet. 68:79-81 (1995), each of which is
incorporated herein by reference). HEK 293 cells are
maintained in DMEM, 25 mM HEPES, 2 mM glutamine,
non-essential amino acids, vitamins, sodium pyruvate,
loo COSMIC CALF SERUM, 100 units/ml penicillin, 100 ug/ml
streptomycin and 0.2 mg/ml 6418 to maintain selection.
Before assaying, cells are washed once with
phosphate buffered saline ("PBS"; without Ca~+ and Mg=') ,
and stripped from the flasks using 0.250 trypsin and
0.5 mM EDTA. Cells are suspended in PBS, 10o COSMIC CALF
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SERUM and 1 mM CaClz. Cell suspensions are prepared at a
density of 2x10' cells/ml for HEK 293 cells expressing
hMCR-3, hMCR-4 or hMCR-5, and 1x10' cellst/ml for HEK 293
cells expressing hMCR-1, Suspensions are placed in a
water bath-ahd allowed to warm tv 37°C for 1 hr.
Binding assays are performed in a total volume
of 250 ~tl for HEK 293 cells. Control and tPSt compounds
are dissolved in distilled water. szSl_HP 467
to (50,000 dpm) (2000 Ci/mmol) (custom labeled by Amersham;
Arlington Heights IL) is prepared in SO mM Tris, pH 7.4,
2 mg/ml BSA, ZO mM CaClz, 5 mM MgClz, 2 mM EDTA and added
to each tube. To each tube is added 4x10 HEK 293 cells
expressing hMCR-3~, rvMCR-4 or hMCR-5, ar 2x10' cells
s5 expressing hMCR-1. Aw ay, are incubated for 2.5 hr at
37°C.
GFJ$ filter plates are prepared by soaking for
at least one hour iri 5 mg/ml BSA and 10 mM CaCl~. Assays
2o are filtered using a Brandel 96-well cell harvested'
(Hrandel Inc,; Gair_hersburg, MD). The filters are washed
four times with cold 50 mM Tris, pH 7_4, and the filter
plates dehydrated for 2 hr and 35 ~.1 of MICROSCINT is
added to each well. Filter plates are counted using a
z5 Parkard Topcount (Packard Instrument Co.) and data
analyzed using GraphPad PRISM v2.0 (GraphPad Software
Inc. ; San Diego CA) and MicrosofC EXCEL v5 . 0a (Microsoft
Corp . ; Red~t~ond WA) .
To assay bicyclic hydantoin derivative
compounds, binding assays are performed in duplicate in a
96 well formaC. HP 467 is prepared in 50 mM Tris, pH
7.4, and 1'SI-HP 467 is diluted to give 100,000 dpm per 50
~1. A bicyclic hydantoin der~.Vative compound,
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is added to the well in 25 u1 aliquots. A 25 ~1 aliquot
of ''='I-HP 467 is added to each well. A 0.2 ml aliquot of
suspended cells is added to each well to give the cell
numbers indicate above, and the cells are incubated at
37°C for 2.5 hr. Cells are harvested on GF/B filter
plates as described above and counted.
EXAMPLE 4
Penile erection due to administration of a bicyclic
hydantoin derivative compound
Adult male rats are housed 2-3 per cage and are
acclimated to the standard vivarium light cycle (12 hr.
light', 12 hr. dark), rat chow and water for a least a
week prior to testing. All experiments are performed
between 9 a.m. and noon and rats are placed in
cylindrical, clear plexiglass chambers during the 60
minute observation period. Mirrors are positioned below
and to the sides of the chambers, to improve viewing.
Observations begin 10 minutes after an
unstraperitoneal injection of either saline or compound.
An observer counts the number of grooming motions,
stretches, yawns and penile erections (spontaneously
occurring, not elicited by genital grooming) and records
them every 5 minutes, for a total of 60 minutes. The
observer is unaware of the treatment and animals are
tested once, with n=6 in each group. Values in the
figures represent the group mean and standard error of
the mean. HP 228 can be used as a positive control for
penile erections. Significant differences between groups
are determined by an overall analysis of variance and the
Student Neunmann-Keuls post hoc test can be used to
identify individual differences between groups
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(p <_ 0.05) .
Although the invention has been described with
reference to the examples provided above, it should be
understood that various modifications can be made by
those skilled in the art without departing from the
invention. Accordingly, the invention is set out in the
following claims.
