Note: Descriptions are shown in the official language in which they were submitted.
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COMBINATORIAL LIBRARIES OF PEPTIDOMIMETIC
MACROCYCLES AND PROCESSES THEREFOR
Field of the Invention
. This Application claims the benefit of U.S. Provisional
Application No. 60/043,532, filed April 11, 1997.
The present invention relates to diverse libraries of
peptidomimetic organopeptide macrocyclic compounds,
combinatorial methods of making such libraries, and an
apparatus for storing and providing a readily accessible
source of such libraries. The apparatus harboring the
present combinatorial libraries is a useful component of
assay systems for identifying compounds for drug
development.
Background of the Invention
Research and development. expenses account for a large
outlay of capital in the pha~__°maceutical industry.
Synthesis of compounds is an expensive and time consuming
phase of research and development. Historically, research
chemists individually synthe:~ized and analyzed high purity
compounds for biological scrE:ening to develop
pharmaceutical leads. Although such methods were
successful in bringing new drugs to the market, the
limitations of individual synthesis and complete compound
characterization considerable slowed the discovery of new
pharmaceutically active compounds. The need for more
rapid and less expensive drug discovery methodology is
increasingly important in today's competitive
pharmaceutical industry.
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Recently, modern drug discovery has utilized
combinatorial chemistry to generate large numbers (102 -
106) of compounds generally referred to as "libraries".
An important objective of combinatorial chemistry is to
generate a large number of novel compounds that can be
screened to identify lead compounds for pharmaceutical
research.
Theoretically, the total number of compounds which
may be produced for a given library is limited only by the
number of reagents available to form substituents on the
variable positions on the library's molecular scaffold.
The combinatorial process lends itself to automation, both
in the generation of compounds and in their biological
screening, thereby greatly enhancing the opportunity and
efficiency of drug discovery.
Combinatorial chemistry may be performed in a manner
where libraries of compounds are generated as mixtures
with complete identification of the individual compounds
postponed until after positive screening results are
obtained. However, a preferred form of combinatorial
chemistry is "parallel array synthesis", where individual
reaction products are simultaneously synthesized, but are
retained in separate vessels. For example, the individual
library compounds can be prepared, stored, and assayed in
separate wells of a microtiter plate, each well containing
one member of the parallel array. The use of standardized
microtiter plates or equivalent apparatus, is advantageous
because such an apparatus is readily accessed by
programmed robotic machinery, both during library
synthesis and during library sampling or assaying.
Combinatorial chemistry can be carried out in
solution phase where both reactants are dissolved in
solution or in solid phase where one of the reactants is
covalently bound to a solid support. Typically,
completion of the solution phase reactions in
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combinatorial chemistry schemes are ensured by selecting
high yielding chemical reactions and/or by using one
reagent in considerable excess. When one reagent is used
in excess, completion of the reaction produces a mixture
of a soluble product with at least one soluble unreacted
reagent. Solid phase synthe:~is offers the advantage that
the solid support-bound products are easily washed free of
excess reagent. Solution ph<~se synthesis typically
requires use of one or more reaction mixture work-up
procedures to separate reaction product from unreacted
excess reagent.
Combinatorial chemistry may be used at two distinct
phases of drug development. In the discovery phase
diverse libraries are created to find lead compounds. In
a second optimization phase, strong lead compounds are
more narrowly modified to find optimal molecular
configurations.
Small peptides as a cla~~s of compounds possess rich
diversity and potent bioactivity and thus remain a vital
starting point for drug discovery. However, due to a
litany of undesirable pharmacokinetic properties of
peptides, peptidomimetics are frequently used to redeploy
important peptide main chain and side chain pharmacophores
onto less peptide-like scaffolds. Peptidomimetics vary
greatly in their resemblance to peptides. Success has
been achieved with peptides containing hydrolytically
stable amide surrogates, mixed organo-peptide frameworks,
and with non-peptide scaffolds alike. Benzodiazepines,
penicillins and cephalosporins are well-established
peptidomimetic drugs and it is easy to see that organo-
peptide chimeras are essential to both rational design and
combinatorial chemistry approaches to drug discovery.
Conformational restriction is important in drug
- pharmacology and efficacy because i) less flexible
molecules usually bind with much higher affinity to their
targets because less energy i;> expended in freezing out
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non-active conformations and ii) conformationally
restricted peptides are usually less susceptible to
digestion by proteases, hence are expected to be more
active drugs. Thus, preparation of cyclic compounds has
been an important drug discovery tactic to take advantage
of those features. [See Hruby, Life Sciences, 31, 189-199
(1982); Kates et al., Tet Lett 34, in 1549-52 (1993);
Kataoka et al., Biooolvmers 32, 1519-33 (1992); Szewczuk
et al., Biochemistry 31, 9132-40 (1992); Schiller et al.,
J. Med Chem., 34 3125-32 (1992); Toniolo, Int. J. Pent
Prot. Res., 35, 287-300 (1990); Veber et al., Nature, 280,
512-4 (1979)]
Barker et al. [J. Med. Chem. 35, 2040-8 (1992)] and
McDowell and Gadek [JACS, 114, 9245-9253, (1992)] have
made constrained cyclic peptides having a single thioether
bond (at the point of ring closure) and bearing the
sequence Arginine-Glycine-Aspartic acid, a high affinity
ligand for glycoprotein IIb-IIIa, an important receptor in
the blood clotting cascade. Wen and Spatola have
synthesized and characterized linear pseudopeptide
libraries bearing 1 to 3 dialkylamino replacements
"[i~rCH2NH]~~ for defined amide bonds [Abstract #60, Division
of Medicinal Chemistry, American Chemical Society 207th
National Meeting in San Diego, CA, March 13-17, 1994.]
For other reports of linear and cyclic peptides bearing a
single thioether replacement for an amide bond, see
Paladino et al., Int. J. Pept Prot Res. 42, 284-93
(1993); Anwer et al., Int. J. Pept Prot Res., 36, 392-9
(1990); Spatola et al., Colloa. INSERM, 174, 2nd Peptide
Forum, 45-54 (1989); and Darlak et al., Pept. Proc Eur
Pent. Sump 20th, 634-6 (1989), June and Gayer, Eds.]
Summary of the Invention
The present invention is directed to the construction
of a series of novel structurally related macrocyclic
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compounds from one or more a~minothioether compounds or one
or more of their corresponding sulfoxide or sulfone
derivatives, collectively referred to herein as "ATAs".
ATA compounds are used in combination with other ATAs and
other organic compounds that bear amine and acid
functionality, to generate the present novel, diverse
macrocyclic compounds having at least 2 sulfur linkages in
the macrocycle ring structure, wherein the oxidation state
of those linkages may vary. By linking ATAs together in
sequence, one may generate an oligomer with a backbone
alternating between sulfur and amide linkages. Moreover,
the flexibility of the present process for preparing the
macrocyclic libraries enables construction of
peptidomimetics that are not constrained to the normal
spacing or "register" found in normal peptides. The
process also enables facile synthesis of macrocycle
libraries having the same number or a different number of
ring atoms of from about 12 to about 40 ring atoms.
Libraries of these macrocyclic ATA compounds are screened
to identify lead compounds through their biological
activity.
In their simplest form, ATAs are dipeptides where a
thioether bridge replaces th~~ central amide linkage, such
as, for example, those described by Spatola, A.F., in
Chemistry and Biochemistry o.f Amino Acids, Peptides and
Proteins, B. VJeinstein, Ed.; Marcel Dekker: New York,
1983; Vol. 7; pages 267-357. A facile synthesis of such
compounds from commercially available amino acids or amino
alcohols and mercapto acids provides a diverse array of
ATA compounds for use in preparation of novel macrocycle
libraries described in the present invention. The diverse
ATA compounds are used in combination with other organic
compounds that bear amine anti acid functionality,
including both proteiogenic ~~mino acids and non-
proteiogenic amino acids, to construct diverse libraries
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of peptidomimetic macrocyclic compounds of the present
invention.
The present invention is also directed to a simple
and direct process for the production of diverse libraries
of novel macrocyclic organo-peptide compounds useful in
the identification of new pharmaceutical lead compounds.
For example, as depicted in Scheme I below, the present
library compounds may be easily prepared using solid phase
synthesis techniques via carbodiimide-mediated coupling of
one or more ATAs with proteiogenic and non-proteiogenic
amino acids, followed by solid phase or solution phase
cyclization by a thioetherforming intramolecular
nucleophilic displacement reaction. The anchor residue
and the terminal bromoacid, as well as the identity,
number and coupling order of the intervening amino acid
and ATA components all contribute to the unique structural
identity of the macrocycle product.
;.>
a' b' c'
FmocHN --> cys --~ ~yS ---- cys
AA1 ATA A
1s 2s 3s 4s
BrA
Cys ~ AA2 Cys
ATA AAA ATA
7s 6s 5s
Scheme I
Steps: a') N-Fmoc-rink amide MBHA resin, 30% piperidine
in DMF, then 5 eq Fmoc-S-trityl-L-cysteine, diisopropyl
carbodiimide/hydroxybenzotriazole (DIC/HOBT) in N-
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methylpyrrolidinone (NMP); ~;teps b') through d') as for
Step a') but with the respective amino acid (AA) or ATA;
e') 30o piperidine in DMF, then 5eq bromoacetic
acid and DIC in NMP;
f') 38:1:1 trifluoroacetic acid (TFA):H20:Et3SiH
for 2 hours, lyophilize, the 2 eq of diisopropylethylamine
in 1:1 CH3CN:H20.
The library is created, stored, and used as an
apparatus comprising a two-dimensional array of
reservoirs, each reservoir containing a predetermined
library reaction product differing from those in adjacent
reservoirs.
Another embodiment of the present invention provides
an assay kit for the identification of pharmaceutical lead
compounds, said kit comprising assay materials and a well
plate apparatus or equivalent apparatus providing a two-
dimensional array of defined reservoirs. The well plate
apparatus provides a diverse combinatorial library,
wherein each well (reservoir) contains a unique macrocycle
compound, or stereoisomers and/or regioisomers thereof.
The well plate apparatus is 'used to provide multiple
reaction zones for making the library, to store the
library and to provide a rea~~ily accessible source of
library compounds.
Brief Description of the Drawincrs
Fig. 1 is a top view of a well plate in accordance
with this invention.
Fig. 2 is a side view o~= a well plate apparatus for
use in the process of this invention.
Detailed Descrit~tion of the Invention
The term "assay kit" as used in accordance with the
present invention refers to an assemblage of two
cooperative elements, namely (1) a well plate apparatus
and (2) biological assay materials.
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"Biological assay materials" are materials necessary
to conduct a biological evaluation of the efficacy of any
library compound in a screen relevant to a selected
disease state.
A "library" is a collection of compounds created
by a combinatorial chemical process, said compounds having
a common scaffold with one or more variable substituents.
A "library compound" is an individual reaction
product, a single compound or a mixture of isomers, in a
combinatorial library.
A "Lead compound" is a library compound in a selected
combinatorial library for which the assay kit has revealed
significant activity relevant to a selected disease state.
A "diverse library" means a library where the
substituents on the combinatorial library scaffold or core
structure, are highly variable in constituent atoms,
molecular weight, and structure, and the library,
considered in its entirety, is not a collection of closely
related homologues or analogues (compare to "directed
library").
A "directed library" is a collection of compounds
created by a combinatorial chemical process, for the
purpose of optimization of the activity of a lead
compound, wherein each library compound has a common
scaffold, and the library, considered in its entirety, is
a collection of closely related homologues or analogues to
the lead compound (compare with "diverse library").
The term "scaffold" as used in accordance with the
present invention comprises a peptidomimetic macrocycle
bearing at least two sulfur linking groups. The scaffold
may be further derivatized using conventional
combinatorial techniques.
"Substituents" are chemical radicals which are bonded
to or incorporated onto the scaffold through the
combinatorial synthesis process. The different functional
groups account for the diversity of the molecules
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throughout the library and are selected to impart
diversity of structure, function and biological activity
to the scaffold in the case of diverse libraries, and
optimization of a particular biological activity in the
a 5 case of directed libraries.
"Reagent" means any chemical compound used in the
combinatorial synthesis process to incorporate
substituents on the scaffold of a library.
"Parallel array synthesis" refers to the method of
conducting combinatorial chemical synthesis of libraries
wherein the individual combinatorial library compounds are
separately prepared and storf=d without prior and
subsequent intentional mixing.
"Simultaneous synthesis"' means making of library
compounds within one production cycle of a combinatorial
method (not making all libra~_y compounds at the same
instant in time).
The "reaction zone" refers to the individual vessel
location where the combinatorial chemical library compound
preparation process of the invention is carried out and
where the individual library compounds are synthesized.
Suitable reaction zones include, but are not intended to
be limited to the individual wells of a well plate
apparatus.
"Well plate apparatus" refers to the structure
capable of holding one or more library compounds in
dimensionally fixed and defined positions.
"Non-interfering substit.uents" are those chemical
radicals that do not significantly impede the process of
the invention and yield stable aminothioether macrocyclic
library compounds.
"Aryl" means one or more aromatic rings, each of 5 or
6 ring carbon atoms and includes substituted aryl having
one or more non-interfering substituents. Multiple aryl
rings may be fused, as in naphthyl, or unfused, as in
biphenyl.
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"Alkyl" means straight or branched chain or cyclic
hydrocarbon having 1 to 20 carbon atoms.
"Substituted alkyl" is alkyl having one or more non-
interfering substituents.
"Halo" means chloro, fluoro, iodo or bromo.
"Heterocycle" or "heterocyclic radical" means one or
more rings of 5, 6 or 7 atoms with or without unsaturation
or aromatic character, optionally substituted with one or
more non-interfering substituents, and at least one ring
atom which is not carbon. Preferred heteroatoms include
sulfur, oxygen, and nitrogen. Multiple rings may be
fused, as in quinoline or benzofuran, or unfused as in 4-
phenylpyridine. Suitable substituents on the heterocyclic
ring structure include, but are not limited to halo, C1-
C1p alkyl, C2-C10 alkenyl, C2-C1p alkynyl, C1-C10 alkoxy,
C~-C12 aralkyl, C~-C12 alkaryl, C1-C10 alkylthio,
arylthio, aryloxy, arylamino, C3-C10 cycloalkyl, C3-C10
cycloalkenyl, di(C1-C10)-alkylamino, C2-C12 alkoxyalkyl,
C1-C6 alkylsulfinyl, C1-C1p alkylsulfonyl, arylsulfonyl,
aryl, hydroxy, hydroxy(C1-C10)alkyl, aryloxy(C1-C10)alkyl,
C1-C10 alkoxycarbonyl, aryloxycarbonyl, C1-C10
alkanoyloxy, aryloyloxy, substituted alkoxy, fluoroalkyl,
nitro, cyano, cyano(C1-C10)alkyl, C1-C1p alkanamido,
aryloylamido, arylaminosulfonyl, sulfonamido, amidino,
carbamido, , carboxy, heterocyclic radical, nitroalkyl,
and -(CH2)m-Z-(C1-C10 alkyl), where m is 1 to 8 and Z is
oxygen or sulfur.
"Organic moiety" means a substituent comprising a
non-interfering substituent covalently bonded through at
least one carbon atom. Suitable substituents onto a
connecting carbon atom include, but are not limited to
hydrogen, halo, C1-C10 alkyl, C2-C10 alkenyl, C2-C10
alkynyl, C1-C10 alkoxy, C~-C12 aralkyl, C~-C12 alkaryl,
C1-C10 alkylthio, arylthio, aryloxy, arylamino, C3-C10
cycloalkyl, C3-C10 cycloalkenyl, di(C1-C1p)-alkylamino,
C2-C12 alkoxyalkyl, C1-C6 alkylsulfinyl, C1-C10
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alkylsulfonyl, arylsulfonyl, aryl, hydroxy, hydroxy(C1-
C1p)alkyl, aryloxy(C1-Cl0)alkyl, C1-C10 alkoxycarbonyl,
aryloxycarbonyl, C1-C10 alkanoyloxy, aryloyloxy,
substituted (C1-C10) alkoxy, fluoroalkyl, nitro, cyano,
cyano(C1-Cl0)alkyl, C1-C10 alkanamido, aryloylamido,
arylaminosulfonyl, sulfonamido, amidino, carbamido,
protected amino, protected carboxy, protected amino,
carboxy, heterocyclic radical, nitroalkyl, and -(CH2)m-Z-
(C1-C10 alkyl), where m is 1 to 8 and Z is oxygen or
sulfur.
The term "amino acid" ass used in accordance with the
present invention includes the 20 proteiogenic amino acids
encoded by the genetic code, as well as hydroxyproline,
alpha-aminoisobutyric acid, sarcosine, citrulline, cysteic
acid, t-butylglycine, t-buty:Lalanine, phenylglycine,
cyclohexylalanine, beta-alan:ine, 4-aminobutyric acid and
other compounds of the general formula:
HN ~! COOH
Rn
wherein Rn is hydrogen or an organic moiety, and Q is an
organic group comprising 1 to 12 carbon atoms and 0 to 4
heteroatoms selected from O, N and S; or Rn taken together
with Q and the nitrogen atom to which they are bound form
a 4 to 7-membered ring.
"Proteiogenic amino acids" are those amino acids of
the formula
CHR1COOH
Rn
wherein R1 is selected from the group consisting of
hydrogen, methyl, isopropyl, isobutyl, sec-butyl,
hydroxymethyl, 1-hydroxyethyl, sulfhydrylmethyl,
2(methylthio)ethyl, benzyl, 4-hydroxybenzyl,
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3-indolylmethyl, carboxymethyl, 2-carboxyethyl,
carbamidomethyl, 2-carbamidoethyl, 4-aminobutyl,
3-guanadinylpropyl and 4-imidazolylmethyl, and Rn is
hydrogen, or Rn and R1 taken together with the common
bonded nitrogen atom form a pyrrolidine ring.
"Activated acid" or "activated acid group" refers to
a carboxylic acid that has been reacted to form a group -
C(O)X wherein X is a leaving group subject to nucleophilic
displacement by nucleophiles. Exemplary of the group X is
chloro (an acid chloride), -OC(O)Rx (an anhydride), or -
ORx (an active ester). Preparation of such activated acid
derivatives and their use in preparation of other acid
derivatives are well known in the art
"Acid reactive groups" refer to those nucleophilic
groups capable of reacting with activated acids to form a
covalent bond. Exemplary of acid reactive groups are -OH,
-SH or -NHRr, where Rr is hydrogen or an organic moiety,
and stabilized carbon anions.
"Solid support" refers to a solvent insoluble
substrate having acid reactive groups for forming
cleavable covalent bonds with acid reagents, such as S-
protected amino-protected mercapto amino acids for use in
preparing the present library compounds.
The term "ring atoms" in defining this invention
refers to those atoms or "ring members" covalently bonded
serially, one to another, to form a ring structure.
A diverse library of macrocyclic compounds is
provided in accordance with the present invention. The
macrocycle library embodied as an apparatus of this
invention serves as a readily accessible source of diverse
macrocyclic compounds for use in identifying new
biologically active macrocyclic compounds through
pharmaceutical and agricultural candidate screening
assays, for use in studies defining structure/activity
relationships, and/or for use in clinical investigation.
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According to the present invention, there is provided
structurally related macrocyclic compounds of the formula
B
. ~C O
Rz Rz
S
2 NH
L
~ L.
C
O
wherein
L is a divalent group o:E the formula 1 lCH2 ) o-1-C6H4CH-~?
or -CHRa- wherein Ra is hydrogen or an organic moiety;
RZ is hydrogen or methy:L;
B is hydroxy, or a group of the formula -NRyRt
wherein Ry is hydrogen, alky=L, aryl or heterocycle, and Rt
is hydrogen, a solid support, or Rt is a group of the
formula
T "_- N E
Ry"
wherein E is hydrogen, a solid support or a
substituent derived from an electrophilic reagent, T is a
divalent linking group, and F:y~~ is hydrogen or a
substituent derived from an electrophilic reagent; or Ry
and Ry~~ taken together with the atoms to which they are
bonded form a 6- to 7-membered ring or a bicyclic or
tricyclic ring comprising 6 to 12 carbon atoms; or Ry or
Ry~~ taken together with -T- and the atoms to which they
are commonly bonded form a 4- to 7-membered ring; or Ry
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and Ry~~ each taken together with T and the nitrogen atom
to which they are respectively bonded form a 5- to 7-
membered ring;
Z is a divalent group of the formula
a(~1)a1-(~2)a2-(ATA1)t1-(AA3)a3-(ATA2)t2-(AA4)a4-
(ATA3 ) t3- (~5 ) a5- (~6 ) a6b
wherein each of AA1-6 is independently a divalent
group of the formula
O
I I
I
Rn
and each of ATA1-3 is independently a divalent sulfur-
linked group of the formula
(~~r, O
II
Rn' Rs
and wherein a1, a2, a3, a4, a5, a6, t1, t2, and t3 are
independently 0 or 1, provided that t1 + t2 + t3 - 1, 2 or
3; and provided further that -Z- is selected to provide a
12- to 40-membered macrocycle ring of Formula I;
wherein in the above formulas for AA1_6 and ATA1-3,
W is a divalent organic group comprising 1 to 12
carbon atoms and 0 to 4 heteroatoms selected from O, N and
S;
Q and Q' are each independently an organic group
comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms
selected from O, N and S;
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m is 0, 1 or 2;
Rs is hydrogen or an or~~anic moiety; and
Rn' is hydrogen, or an organic moiety, or Rn taken
together with Q and the atom: to which they are bonded
form a 4- to 7-membered ring,;
Rn~ and Rs taken together with -Q'- and the atoms to
which they are bonded form a 4- to 7-membered ring or a
bicyclic or tricyclic ring comprising 6 to 12 carbon
atoms; or
Rn~ or Rs taken together with -Q'- and the atoms to
which they are commonly bonded form a 4- to 7-membered
ring.
The library compounds of this invention have a
molecular weight of about 200 to about 1500, more
typically about 250 to about 1250. The present macrocycle
library compounds as represented by Formula I include 12
to 40 ring atoms. In one emb~~diment the library compounds
are constructed to have 12 to 24 ring atoms, at least two
of which are sulfur. The rin~~ atoms are derived from four
types of reactants during the solid phase synthesis: 1) a
solid support-anchored orthogonally protected
trifunctional (protected thio:L, protected amino, carboxy)
compound (viz. cysteine or penicillamine); 2) aminoacids,
selected from proteiogenic amino acids and non-
proteiogenic amino acids; 3) ATAs (aminothioether acids
and their corresponding sulfo~s:ide and sulfone
derivatives); and 4) alpha-halo acids, preferably alpha-
bromo acids or carboxy- or carboxymethyl-substituted
benzyl bromides. Generally, m.acrocycle synthesis is
initiated by deprotecting the support anchored amine
functional group. The linear (or at least acyclic)
macrocycle precursor is synthesized by serial peptide
coupling of at least one ATA a:nd zero, one or more amino
acids. The resulting terminal amine functionality is
finally coupled, by peptide bond formation, to an alpha-
halo acid or a carboxy/carboxymethyl substituted benzyl
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bromide to provide an electrophilic (bromomethyl)
terminus. Upon deprotection of the support- bound thiol
and treatment with a non-nucleophilc base, the acyclic
precursor undergoes intramolecular thioether-forming
cyclization. Accordingly, the support bound orthogonally
protected cysteine or penicillamine contributes 4 ring
atoms to the macrocycle structure, use of an alpha-halo
acid contributes two ring atoms, and use of a
carboxy/carboxymethyl substituted benzyl bromide
contributes 4 to 7 ring atoms to the macrocycle structure.
The remaining ring members are determined by the nature,
coupling order and number of the amino acids and ATAs
coupled during synthesis of the acyclic support bound
macrocycle precursor.
The libraries in accordance with this invention can
be synthesized to have diversity in the number of ring
atoms, or they can be prepared so that each member of the
library has the same number of ring atoms with diversity
in the library being introduced by the nature and coupling
order of the component amino acids (AA1-6) and ATAs.
Additional elements of diversity can be introduced into
the present macrocycle library by the preparation and use
of solid supports having selected acid reactive groups
such as, for example, those acid reactive groups derived
from reaction of diamines with p-nitrophenylcarbonate
Wang resin. Trifluoroacetic acid mediated cleavage of the
synthesized macrocycles or acyclic macrocycle precursors
from such diamine reacted resin provides a "side chain"
having amine functionality that can be reacted with a wide
variety of electrophilic reagents.
In one embodiment of the invention there is provided
a diverse library of macrocycle compounds of Formula I
wherein B is NH2. Such library compounds are derived from
the corresponding library compounds wherein B is a solid
support, particularly a Rink-Amide AM resin or Rink-Amide
MBHA resin by trifluoracetic acid mediated cleavage of the
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covalently bound library compound or its acyclic precursor
from the solid support.
Another embodiment of the present invention provides
macrocycle library compounds of Formula I wherein B is a
group of the formula -NRyRt wherein Ry is hydrogen, alkyl,
aryl or heterocycle and Rt is hydrogen, a solid support,
or Rt is a group of the formula
'I' --- N E
Ry"
wherein T, Ry~~ and E are as defined above. The compounds
where E is hydrogen can be d~srived from corresponding
library compounds wherein E :is a solid support,
particularly a Wang resin, b~~r trifluoroacetic acid
mediated cleavage, or they c<~n be derived by cyclization
of the corresponding acyclic polymers. The library
compounds of Formula I wherein E is hydrogen are useful as
core structures for the synthesis of directed macrocycle
libraries having diversity in the group E through
reactions with various electrophilic reagents to construct
variable "side chain" functionality on the macrocycle
library compounds.
The library compounds of: Formula I wherein the group
B or E is a covalently bonded solid support (and the
corresponding acyclic precur:~or compounds) also represent
embodiments of the present irwention. A general
compilation of solid support~~ (resins) can be found in
"Supports for Solid Phase Organic Synthesis," Martin
Winter, pp 465-510, in Combinatorial Peptide and Non-
peptide Libraries, edited by Gunther Jung (1996), VCH
Publishers (Weinheim, Germany). Many solid supports
having acid reactive groups, such as hydroxy and primary
or secondary amino groups (or precursors thereto) are
commercially available. The acid reactive groups
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covalently bound to solid supports are typically designed
to have selectively cleavable covalent bonds linking it to
the solid support. Illustrative of commercially available
solid supports include, but are not intended to be limited
to, the following resins:
Wang resin (where the sphere represents commercial
divinylbenzene-crosslinked polystyrene):
Wang resin functionalized with various amino acids (many
varieties are commercially available):
~a~R
NIHFawo
Wang p-nitrophenylcarbonate resin:
NO=
0
~o
Rink Amide AM resin:
FmooHN OMo
H I
N' ~
OMo
O
O
Rink Amide MBHA resin:
FmeWIH OIM
O OAAo
H O
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_.1g-
Knorr Resin:
_b ~ I
H ~ 1 ~ G 0W
The library compounds cf Formula I of this invention
wherein B is the group -NRyRt, Rt is a group of the
formula
T' N E
Ry
and wherein T and Ry~~ are as defined above and E is a
solid support, are derived from the modified Wang resin
bearing acid reactive groups of the formula
O
HN T N CO-CH2-CsH4-O-CHZ-C6Hd--Polymer
Ry Ry"
wherein Ry, T, and Ry~~ are as defined above. The
modified Wang resin is prepared by reacting Wang
p-nitrophenylcarbonate resin with an excess of diamine of
the formula
HI T IH
Ry Ry"
Exemplary of diamine compounds suitable for preparing
modified Wang resins for use in preparing library
compounds of this invention wherein E is a solid support
or hydrogen (after cleavage) include, but are not intended
to be limited to the following:
1, 2 -DIAMINOCYCLOHEX,D.NE
1,3-BIS(AMINOMETHYL)CYCLOHEXANE
D-CYSTINE
L-ARGININE
2,6-DIAMINOPIMELIC i~CID
2,6-DIAMINOPIMELIC ~~CID
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N-EPSILON-CBZ-L-LYSINE
N-EPSILON-ACETYL-L-LYSINE
N-(3-AMINOPROPYL)CYCLOHEXYLAMINE
ETHYLENEDIAMINE-N, N'-DIACETIC
ACID
N,N'-DIBENZYLETHYLENEDIAMINE
PIPERAZINE
2-METHYLPIPERAZINE
2,6-DIMETHYLPIPERAZINE
1-(2-AMINOETHYL)PIPERAZINE
4,4'-BIPIPERIDINE DIHYDROCHLORIDE
4-(AMINOMETHYL)PIPERIDINE
1,4-BIS(3-AMINOPROPYL)PIPERAZINE
HOMOPIPERAZINE
ACETALDEHYDE-AMMONIA TRIMER
ACETALDEHYDE-AMMONIA TRIMER
M-XYLYLENEDIAMINE
1,10-DIAMINODECANE
1,12-DIAMINODODECANE
N-ISOPROPYLETHYLENEDIAMINE
N-ETHYLETHYLENEDIAMINE
TETRAETHYLENEPENTAMINE
TRIETHYLENETETRAMINE
2-(2-AMINOETHYLAMINO)ETHANOL
DIETHYLENETRIAMINE
N-(N-PROPYL)ETHYLENEDIAMINE
N-(2-AMINOETHYL)-3-
AMINOPROPYLTRIMETHOXYSILANE
TRIS(2-AMINOETHYL)AMINE
N-(2-AMINOETHYL)-1,3-
PROPANEDIAMINE
3,3'-IMINOBISPROPYLAMINE
1,4-DIAMINOBUTANE
1,5-DIAMINOPENTANE
1,6-HEXANEDIAMINE
1,7-DIAMINOHEPTANE
1,8-DIAMINOOCTANE
1,9-DIAMINONONANE
N,N'-DIETHYL-2-BUTENE-1,4-DIAMINE
P-XYLYLENEDIAMINE
D-ARGININE MONOHYDROCHLORIDE
(+/-)-3-AMINOPIPERIDINE
DIHYDROCHLORIDE
2,2'-OXYBIS(ETHYLAMINE)
DIHYDROCHLORIDE
CYSTAMINE DIHYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE
D-LYSINE MONOHYDROCHLORIDE
AGMATINE SULFATE
2-METHYL-1,5-DIAMINOPENTANE
ISOPHORONEDIAMINE
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2-N-BUTYLAMINOETHS'LAMINE
1,4-DIAMINOCYCLOHE~XANE
S-(2-AMINOETHYL)-I~-CYSTEINE
HYDROCHLORIDE
N,N-DIETHYLDIETHYL~ENETRIAMINE
TERT-BUTYL N-(6-
AMINOHEXYL)CARBAMP,TE
HYDROCHLORIDE
1,4-BIS(AMINOMETHYL)CYCLOHEXANE
1,8-DIAMINO-3,6-DIOXAOCTANE
N-BENZYLETHYLENEDIAMINE
PIPERAZINE-2-CARBOXYLIC ACID
DIHYDROCHLORIDE
L-CYSTINE DIHYDROXAMATE
L-ARGININE HYDROXAMATE
HYDROCHLORIDE
DIAMINOBIOTIN
L-LYSINE HYDROXAMATE
HYDROCHLORIDE
L-LYSINAMIDE DIHYDROCHLORIDE
3-AMINOPYRROLIDINE
DIHYDROCHLORIDE
(2R,2R)-(-)-1,2-
DIAMINOCYCLOHEXANE
(1S, 2S) - (+) -1, 2-
DIAMINOCYCLOHEXANE
CIS-1,2-DIAMINOCYC:LOHEXANE
TRAMS-1,2-DIAMINOC'i~CLOHEXANE
L-CYSTINE
L-LYSINE
L(-)-ALPHA-AMINO-El?SILON-
CAPROLACTAM
L-ARGININE MONOHYDROCHLORIDE
L-ARGININE MONOHYDROCHLORIDE
L-2,4-DIAMINOBUTYRIC ACID
DIHYDROCHLORIDE
L-ORNITHINE MONOHYI)ROCHLORIDE
L-ORNITHINE MONOHYI)ROCHLORIDE
DL-LYSINE MONOHYDROCHLORIDE
DL-LYSINE MONOHYDROCHLORIDE
L-LYSINE MONOHYDROC'_HLORIDE
DL-ORNITHINE MONOHYDROCHLORIDE
L-LYSINE DIHYDROCHI~ORIDE
DL-HOMOCYSTINE
TRAMS-2,5-DIMETHYLF'IPERAZINE
ETHAMBUTOL DIHYDROC.'HLORIDE
(1S,2S)-(-)-1,2-
DIPHENYLETHYLENEDIp,MINE
(2R,2R)-(+)-1,2-
DIPHENYLETHYLENEDIP,MINE
DL-CYSTINE
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1,3-DIAMINOPENTANE
(4S,5S)-4,5-DI(AMINOMETHYL)-2,2-
DIMETHYLDIOXOLANE
PIPERAZINE HEXAHYDRATE
(S)-(+)-2-METHYLPIPERAZINE
(S) - (+) -2-
(AMINOMETHYL)PYRROLIDINE
N1-ISOPROPYLDIETHYLENETRIAMINE
(R)-(-)-2-METHYLPIPERAZINE
N-PHENYL-4-PIPERIDINAMINE
2-AMINO-6-FLUOROBENZYLAMINE
2-AMINOBENZYLAMINE
4-AMINOBENZYLAMINE,
and the like.
In accordance with another embodiment of this
invention there is provided a library of macrocycle
compounds of Formula I wherein the divalent group Z is
selected so that a1 + a2 + a3 + a4 + a5 + a6 = 0 to 6 in
integral steps. In other words, one to six amino acids
can be integrated into the macrocycle ring. In a related
embodiment of the invention the library compounds are also
defined so that t1 + t2 + t3 - 1, 2 or 3. Illustrative of
the divalent group Z in those embodiments include, but are
not intended to be limited to:
a ATA1 _b,
a AA1-ATA1-ATA2-AA4 b,
a ATA1-AA3-ATA2 b,
a AA1-ATA1-ATA2-AAg-AA5 b,
~ ATA1-AA3-AA4-ATA3 b,
a ATA1-ATA2-ATA3-AA5 b,
a AA1-ATA2-ATA3-AA5-AA6 b,
a AA3-ATA2-AAg b,
a ATA2-AA4-ATA3-AA5 b,
a AA1-ATA1-ATA2-ATA3 b,
a AA4-ATA3-AA5-AA( b,
~ AA1-ATA1 b,
2 5 a ATA1-AA3 b, and
a ATA1-AA3-ATA2-ATA3 b .
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One preferred embodiment of the invention is a library of
macrocyclic compounds of the formula
B
- C O
S
NH
L
z
C -
O
wherein
L is a divalent group of the formula 1 (CHZ) o_1-C6H9CH2?
or -CHRa- wherein Ra is hydrogen or an organic moiety;
RZ is hydrogen or methyl;
B is hydroxy or a group of the formula -NRyRt wherein
Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is
hydrogen, a solid support, or Rt is a group of the formula
T -"--. N E
Ry ~~
wherein E is hydrogen, a solid support or a
substituent derived from an electrophilic reagent, T is a
divalent linking group, and R.~,~~ is hydrogen or a
substituent derived from an e:Lectrophilic reagent; or Ry
and Ry~~ taken together with the atoms to which they are
bonded form a 4- to 7-memberec3 ring or a bicyclic or
tricyclic ring comprising 6 to 12 carbon atoms; or Ry or
Ry~~ taken together with -T- and the atoms to which they
are commonly bonded form a 6- to 7-membered ring or Ry and
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Ry~~ each taken together with T and the nitrogen atom to
which they are respectively bonded form a 5- to 7-membered
ring;
Z is a divalent group of the formula
a(AA1)-(ATA1)-(AA3)~
wherein AA1 and AA3 are independently a divalent
group of the formula
O
Q C
Rn
and
ATA is a divalent sulfur group of the formula
O
N-Q'-CH-S W C
Rn' RS
wherein W is a divalent organic group comprising 1 to
12 carbon atoms and 0 to 4 heteroatoms selected from the
group consisting of O, N and S;
Q and Q' are each independently an organic group
comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms
selected from O, N and S;
m is 0, 1 or 2;
RS is hydrogen or an organic moiety; and
Rn' is hydrogen, or an organic moiety, or Rn taken
together with Q and the atoms to which they are bonded
form a 4- to 7-membered ring;
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Rn~ and Rs taken together with -Q'- and the atoms to
which they are bonded form a 4- to 7-membered ring or a
bicyclic or tricyclic ring comprising 6 to 12 carbon
atoms; or
- 5 Rn~ or Rs taken together with -Q'- and the atoms to
which they are commonly bonded form a 4- to 7-membered
- ring.
Another embodiment of this invention provides a
library of macrocycle compounds of Formula I wherein each
amino acid used to form the macrocycle is a proteiogenic
amino acid.
Still another embodiment: of this invention is
directed to a library of macrocycle compounds of the
Formula I above wherein Q and/or Q~ is a group of the
formula
- C -_ _.- CH 2 - CH
R1 R2 0
r 1
wherein R1 is hydrogen, or a non-interfering
substituent; and
R2 is hydrogen or an organic moiety; or
R1 taken together with Rn forms a 4- to 7-membered
ring, or R1 taken together with Rs forms a 4- to 7-
membered ring; or R1 taken together with R2 forms a 3- to
6-membered ring.
In still another embodiment there is provided a
diverse library of macrocycle compounds of Formula I
wherein W is a divalent organic group selected from the
group consisting of -CHRg-, -~~R4R5CR6R~-, and a 5- or 6-
membered aromatic ring containing 0 to 4 heteroatoms
selected from O, N and S;
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wherein Rg is hydrogen or an organic group;
R4, R5, and R6 are independently hydrogen or alkyl;
and
R~ is hydrogen, hydroxy, protected hydroxy, amino or
protected amino or substituted amino wherein the
substituent is derived from an electrophilic group.
The present library compounds can be prepared using
combinatorial synthesis protocols, or individual
macrocycle library compounds can be prepared by standard
chemical synthesis techniques and used as core structure
for preparation of directed libraries.
Further in accordance with this invention there is
provided a process for preparing a combinatorial library
of compounds of the formula
B
~C O
Rz z
S
NH
L a/
bb
~~z
O
wherein L, Z, RZ and B are as defined above. The process
comprises the steps of:
(a) covalently bonding an amino-protected, thiol-
protected starting material of the formula
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-:27-
RZ RZ
COOH
PSS
I~THP
wherein RZ is defined as abo~re, and PS and P are thiol- and
amino-protecting groups, respectively, to a solid support
including acid reactive groups;
(b) removing the amino protecting group on the solid
support bound starting mater~_al;
(c) coupling the resulting solid support bound thio-
protected starting material with an amino-protected amino
acid of the formula
p N -"-" Q C OOH
Rn
or an amino-protected compound of the formula
~~~m
P IV Q' CH S W COOH
Rd Rs
wherein in said formulas P is an amino-protecting group
and, Rn, Rn~, Q, Q', Rs~ m and W are as defined above;
(d) removing the amino-;protecting group from the
coupled solid support bound product;
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(e) optionally repeating step (c) and step (d) one
or more times using the same or a different amino-
protected amino acid or amino-protected thioether acid
(f) coupling the solid support bound product with an
organic acid of the formula
1
COON
2
X
wherein L is as defined above and X is a leaving group
subject to nucleophilic displacement;
(g) removing the thiol-protecting group to enable
macrocycle formation by nucleophilic displacement of the
group X and cleaving the covalently bound product from the
solid support; and
(h) when the acid reactive group on the solid
support is a covalently bound group of the formula
O
HN T N CO-CH2-C6H~0-CHZ-C6H,~Polymer
Ry Ry
optionally reacting the resulting product of Formula I
wherein B is a group of the formula
N T NH
2 5 Ry Ry ,.
with an amine reactive organic electrophilic agent;
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provided that at least one of the steps (c) and (e)
is carried out using an amino-protected aminothioether
acid.
Each of the reaction steps, whether conducted on
solid phase or in solution phase, is carried out using a
dry, inert, polar, aprotic solvent at a temperature of
about 0° to about 30°C. In the case of solution phase
reactions, stoichiometricall:y equivalent amounts or near
stoichiometric amounts of reactants are typically
employed. The use of solid :phase synthesis for most of
the process steps (the cyclization step can be conducted
in solid phase or solution phase) allows the flexibility
of using excess reagents to ~~ptimize yield without
complicating product isolation. Using solid phase
synthesis techniques, effective work-up to remove any
excess reagents comprises wa:~hing the solid support bound
product one or more times wit=h reaction solvent. Suitable
solvents for carrying out the process steps include
dimethylformamide, dimethylsulfide, tetahydrofuran,
N-methylpyrrolidone, dioxane, ethyl acetate, diethyl ether
and the like. The reaction :steps can be carried out using
standard combinatorial chemistry protocols to produce
arrays of the present library. compounds, or they can be
carried out on larger scale using standard chemical
synthesis, work-up, and product isolation and purification
procedures.
The initial step in the process for preparation of
the present library compounds. of formula I comprises
covalently bonding to a solid support an amino-protected,
thiol-protected starting material of the formula
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RZ RZ
COOH
PSS
NHP
wherein Rz is defined as above, and Ps 'and P are thiol-
protecting and amino-protecting groups, respectively,
wherein the solid support includes covalently bound acid
reactive groups (e. g. hydroxy or primary or secondary
amino). Suitable thiol-protecting groups include, but are
not intended to be limited to, trityl(triphenylmethyl),
4-methoxyphenyl-diphenylmethyl, di(4-
methoxyphenyl)phenylmethyl, and the like. Exemplary of
monovalent amino-protecting groups include, but are not
intended to be limited to:
9-fluorenylmethyloxycarbonyl (Fmoc),
tert-butyloxycarbonyl (Boc),
allyloxycarbonyl (Alloc),
2-trimethylsilylethoxycarbonyl (Teoc),
biphenylisopropyloxycarbonyl (Bpoc),
nitroveratryloxycarbonyl (Nvoc),
(4-methoxyphenyl)diphenylmethyl (MMTr),
(4,4'-dimethoxyphenyl)phenylmethyl (DMT) and
benzyloxycarbonyl (Cbz or Z),
and the like.
Reaction conditions for the preparation of protected
amines utilizing such protecting groups and conditions for
the selective removal of such groups to provide the
corresponding amines are well-known in the art. See for
example, "Protective Groups in Organic Synthesis" 2nd ed.,
by Theodora W. Greene and Peter G.M. Wuts (1991), John
Wiley and Sons (New York).
Covalent coupling of orthogonally protected cysteine
or penicillamine to the solid support is carried out by
forming an activated form of the acid, such as an active
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ester, and reacting the activated form with the solid
support, typically using an excess (2-10 fold) of the
activated form to optimize yield of the support-coupled
product. Excess reagent is 'washed from the reacted solid
support.
The support-coupled product is then reacted to remove
amino-protecting groups using the protecting group
dependent cleavage conditions. Thus, for example, the
fluorenylmethyloxycarbonyl group, a preferred amino-
protecting group for solid phase synthesis, is removed by
treatment of the solid support bound product with from
about 20% to about 30o piper:idine in dimethylformamide.
The resulting solid support bound thio-protected
starting material is then coupled with an amino-protected
amino acid of the formula
p N '-"' Q "- COOH
Rn
or an amino-protected ATA of the formula
~~~m
Q' CH- S W COOH
R~, RS
wherein P is an amino-protecting group and Rn, Rn~, Q, Q~
Rs, m and W are as defined above. The coupling reaction
is carried out under conditions paralleling those
described above for coupling the orthogonally protected
cysteine or penicillamine to the solid support using a 2-
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fold excess of an activated form of the protected amino
acid or ATA.
Any of a wide variety of commercially available or
readily synthesized amino acids can be employed in this
5 step. The protected amino acids typically have a
molecular weight of about 75 to about 800. Exemplary of
such amino acid reactants (and protected derivatives
thereof) suitable for use in this process include, but are
not intended to be limited to, the following:
1-AMINO-1-CYCLOPENTANECARBOXYLIC ACID
1-AMINO-1-CYCLOHEXANECARBOXYLIC ACID
N-ALPHA-CBZ-L-ARGININE
N-ALPHA-BENZOYL-L-ARGININE
CHLOROACETYL-L-TYROSINE
DL-M-TYROSINE
L-DOPA
4-BROMO-DL-PHENYLALANINE
DL-4-FLUOROPHENYLALANINE
DL-4-CHLOROPHENYLALANINE
O-METHYL-L-TYROSINE
O-BENZYL-L-TYROSINE
L-TYROSINE
3-IODO-L-TYROSINE
S-(TERT-BUTYLTHIO)-L-CYSTEINE HYDRATE
D-CYSTINE
S-TRITYL-L-CYSTEINE
S-METHYL-L-CYSTEINE
S-BENZYL-L-CYSTEINE
S-CARBOXYMETHYL-L-CYSTEINE
LANTHIONINE
L-ASPARTIC ACID
L-LEUCINE
DL-HOMOSERINE
L-HOMOPHENYLALANINE
DL-METHIONINE SULFOXIDE
L-METHIONINE SULFOXIMINE
D-METHIONINE
L-ETHIONINE
L-C-ALLYLGLYCINE
GAMMA-L-GLUTAMYL-L-GLUTAMIC ACID
L-GLUTAMIC ACID 5-METHYL ESTER
L-GLUTAMIC ACID GAMMA-BENZYL ESTER
L-GLUTAMIC ACID
L-ARGININE
L-ALPHA-AMINOADIPIC ACID
2,6-DIAMINOPIMELIC ACID
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_ ?~ 3 _
N-EPSILON-CBZ-L-LYSINE
N-EPSILON-ACETYL-L-LYSINE
N-CBZ-L-ALANINE
Z-L-SERINE
N-(TERT-BUTOXYCARBONYL)-L-PHENYLALANINE
5-BROMO-N-{CARBOXYMETHYL)ANTHRANILIC ACID
N-(4-HYDROXYPHENYL)GLYCINE
N-(TERT-BUTOXYCARBONYL)GLYCINE
N-CBZ-GLYCINE
HIPPURIC ACID
2-IODOHIPPURIC ACID
2-HYDROXYHIPPURIC ACID
Z-L-ASPARTIC ACID
L-ASPARTYL-L-PHENYLALANINE METHYL ESTER
N-CBZ-L-GLUTAMIC ACID
N-ACETYL-L-GLUTAMIC ACID
DL-ALPHA-METHYLTYROSINE
N-CARBOBENZYLOXY-2-METHYLAI~ANINE
2-(METHYLAMINO)ISOBUTYRIC ACID
ALPHA-HYDROXYHIPPURIC ACID
ETHYLENEDIAMINE-DI(0-HYDROXYPHENYLACETIC ACID)
D-(-)-p-HYDROXYPHENYLGLYCI1'dE
D-PENICILLAMINE DISULFIDE
D-TERT-LEUCINE
DL-DOPS
DL-VALINE
DL-ISOLEUCINE
D-SERINE
D-PHENYLALANINE
DL-2-FLUOROPHENYLALANINE
6-HYDROXYDOPA
DL-3-FLUOROPHENYLALANINE
N-ACETYLGLYCINE
N-TRITYLGLYCINE
TRICINE
GUANIDOACETIC ACID
SARCOSINE
IMINODIACETIC ACID
ETHYLENEDIAMINE-N, N'-DIACE'I'IC ACID
D-ALLO-THREONINE
N-CBZ-S-BENZYL-L-CYSTEINE
N-ACETYL-DL-PENICILLAMINE
DL-PENICILLAMINE
DL-CYSTEINE
DL-HOMOCYSTEINE
L-AZETIDINE-2-CARBOXYLIC ACID
L-CARNOSINE
DL-HISTIDINE
L-THIAZOLIDINE-4-CARBOXYLIC ACID
3,4-DEHYDRO-DL-PROLINE
3-AMINOPYRAZOLE-4-CARBOXYLIC' ACID
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DL-PROLINE
CIS-4-HYDROXY-D-PROLINE
DL-ALPHA-(2-THIENYL)GLYCINE
BETA-2-THIENYL-DL-ALANINE
L-ABRINE
D-TRYPTOPHAN
5-BROMO-DL-TRYPTOPHAN
5-FLUORO-DL-TRYPTOPHAN
DL-5-METHOXYTRYPTOPHAN
DL-5-HYDROXYTRYPTOPHAN
5-METHYL-DL-TRYPTOPHAN
6-FLUORO-DL-TRYPTOPHAN
1-METHYL-DL-TRYPTOPHAN
L-PIPECOLINIC ACID
NIPECOTIC ACID
ISONIPECOTIC ACID
NITRO-L-ARGININE
3-NITRO-L-TYROSINE
4-NITROHIPPURIC ACID
DL-HOMOCYSTEIC ACID
P-AMINOOXANILIC ACID
4-AMINOHIPPURIC ACID
ALBIZZIIN
DL-CITRULLINE
S-CARBAMYL-L-CYSTEINE
N-ALPHA-CBZ-L-ASPARAGINE
D-ASPARAGINE
N-CBZ-L-GLUTAMINE
L-GLUTAMINE
2,2-DIPHENYLGLYCINE
2-AMINOISOBUTYRIC ACID
D-(-)-ALPHA-PHENYLGLYCINE
DL-3-AMINO-3-PHENYLPROPIONIC ACID
D-ALANINE
DL-3-AMINOBUTYRIC ACID
DL-2-AMINOBUTYRIC ACID
D-NORVALINE
D-NORLEUCINE
DL-2-AMINO-N-CAPRYLIC ACID
DL-2-AMINO-N-CAPRYLIC ACID
GLYCINE
DL-ISOSERINE
DL-4-AMINO-3-HYDROXYBUTYRIC ACID
DL-3-AMINOISOBUTYRIC ACID
BETA-ALANINE
4-AMINOBUTYRIC ACID
5-AMINOVALERIC ACID
N-ALPHA-ACETYL-L-LYSINE
Z-L-VALINE
N-(4-NITROBENZOYL)-BETA-ALANINE
N-(4-AMINOBENZOYL)-BETA-ALANINE
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1-AMINOCYCLOPROPANE-1-CARBOXYLIC ACID
(R)-(-)-N-(3,5-DINITROBEN~~OYL)-ALPHA-PHENYLGLYCINE
N-(3,5-DINITROBENZOYL)-DL--LEUCINE
(+/-)-3-AMINOADIPIC ACID
ALPHA-METHYL-DL-PHENYLALANINE
DL-2-AMINO-5-PHOSPHONOVALE;RIC ACID
DL-3-HYDROXYNORVALINE HYDF;ATE
(+/-)-INDOLINE-2-CARBOXYLIC ACID
BETA-(2-THIAZOLYL)-DL-ALANINE
L-GAMMA-CARBOXYGLUTAMIC ACID
L-2-(BENZYLOXYCARBONYLAMInfO)-4-SULFAMOYLBUTYRIC ACID
L-2-AMINO-4-SULFAMOYLBUTYR.IC ACID
1-AMINO-1-CYCLOPROPANECAREOXYLIC ACID HYDROCHLORIDE
SARCOSINE HYDROCHLORIDE
4-(METHYLAMINO)BUTYRIC ACID HYDROCHLORIDE
L-TYROSINE HYDROCHLORIDE
L-GLUTAMIC ACID HYDROCHLORIDE
D-ARGININE MONOHYDROCHLORIDE
N-EPSILON-METHYL-L-LYSINE HYDROCHLORIDE
L-HOMOARGININE HYDROCHLORIDE
2-AMINO-3-PHENYLBUTANOIC ACID HYDROCHLORIDE
(+/-)-1,2,3,4-TETRAHYDRO-3-ISOQUINOLINECARBOXYLIC
ACID HYDROCHLORIDE
DL-PIPECOLINIC ACID HYDROCHLORIDE
5-AMINOLEVULINIC ACID HYDROCHLORIDE
GLYCINE HYDROCHLORIDE
5-HYDROXY-DL-LYSINE HYDROCI~iLORIDE
DL-2,3-DIAMINOPROPIONIC ACID HYDROBROMIDE
DL-2,3-DIAMINOPROPIONIC ACID MONOHYDROCHLORIDE
DL-.2,4-DIAMINOBUTYRIC ACID DIHYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE
5-AMINOVALERIC ACID HYDROCHLORIDE
D-LYSINE MONOHYDROCHLORIDE
DL-2-AMINO-4-PHOSPHONO BUTYRIC ACID
DL-2-AMINO-4-PHOSPHONO BUTYRIC ACID
N-PHENYLGLYCINE
DL-2-AMINO-3-PHOSPHONOPROP7_ONIC ACID
N-CBZ-L-PHENYLALANINE
N-(CARBOXYMETHYL)ANTHRANIL7:C ACID
N-(P-TOLUOYL)-GLYCINE
L-ASPARTIC ACID MONOPOTASSI:UM SALT
L-ASPARTIC ACID MONOPOTASSI:UM SALT
N-CARBOBENZYLOXY-L-LEUCINE
N-CARBOBENZYLOXY-L-ISOLEUCI:NE
3-AMINO-2,3-DIHYDROBENZOIC ACID HYDROCHLORIDE
FMOC-L-VALINE
N-ALPHA-FMOC-L-ISOLEUCINE
FMOC-L-TRYPTOPHAN
N-ALPHA-FMOC-L-PHENYLALANINE
N-ALPHA-FMOC-L-ASPARAGINE
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FMOC-L-LEUCINE
FMOC-L-METHIONINE
N-ALPHA-FMOC-L-GLUTAMINE
FMOC-GLYCINE
N-(TERT-BUTOXYCARBONYL)-L-TYROSINE
N-EPSILON-BOC-L-LYSINE
N-(TERT-BUTOXYCARBONYL)-L-ALANINE
N-(TERT-BUTOXYCARBONYL)-L-SERINE
N-(P-TOLUENESULFONYL)-L-PHENYLALANINE
STATINE
BOC-L-ASPARTIC ACID
N-ALPHA-BOC-L-ASPARAGINE
N-ALPHA-BOC-L-LYSINE
N-ALPHA-CBZ-L-LYSINE
N-(TERT-BUTOXYCARBONYL)-L-ISOLEUCINE
N-(TERT-BUTOXYCARBONYL)-D-METHIONINE,
DICYCLOHEXYLAMMONIUM SALT
BOC-L-ARGININE
DL-(2-FLUOROPHENYL)-GLYCINE
L-ALPHA-(2-AMINOETHOXYVINYL)GLYCINE HYDROCHLORIDE
N-P-TOSYLGLYCINE
BETA-GUANIDINOPROPIONIC ACID
N-(PHOSPHONOMETHYL)-GLYCINE
N-ALPHA-FMOC-D-TRYPTOPHAN
DL-3-(3,4-DIHYDROXYPHENYL)ALANINE
4-CHLORO-L-PHENYLALANINE
P-IODO-D-PHENYLALANINE
O-PHOSPHO-DL-TYROSINE
D-TYROSINE
DL-TYROSINE
3-FLUORO-DL-TYROSINE
D-ASPARTIC ACID
DL-ASPARTIC ACID
DL-LEUCINE
D-LEUCINE
L-HOMOSERINE
D-HOMOPHENYLALANINE
DL-HOMOPHENYLALANINE
DL-METHIONINE
L-METHIONINE
D-ETHIONINE
DL-ETHIONINE
DL-2-AMINO-4-PENTENOIC ACID
D-GLUTAMIC ACID
D-ARGININE
D-ALPHA-AMINOADIPIC ACID
DL-ALPHA-AMINOADIPIC ACID
N-CBZ-DL-ALANINE
N-CARBOBENZOXY-DL-SERINE
DL-THREONINE
L-ALPHA-METHYLTYROSINE
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L-TERT-LEUCINE
D-VALINE
L-VALINE
D-ISOLEUCINE
L-ISOLEUCINE
DL-SERINE
L-5ERINE
DL-PHENYLALANINE
L-PHENYLALANINE
L-CYSTINE
L-ALLO-THREONINE
D-THREONINE
L-THREONINE
D-PENICILLAMINE
L-PENICILLAMINE
L-CYSTEINE
L-HISTIDINE
D-PROLINE
L-PROLINE
CIS-4-HYDROXY-L-PROLINE
L-HYDROXYPROLINE
BETA-(2-THIENYL)-L-ALANINE
DL-TRYPTOPHAN
L-TRYPTOPHAN
L-5-HYDROXYTRYPTOPHAN
D-PIPECOLINIC ACID
DL-PIPECOLINIC ACID
L-CITRULLINE
L-ASPARAGINE
DL-ALPHA-PHENYLGLYCINE
L(+)-ALPHA-PHENYLGLYCINE
DL-ALANINE
L-ALANINE
(R)-(-)-2-AMINOBUTYRIC ACID
(S)-(+)-2-AMINOBUTYRIC ACID
DL-NORVALINE
L-NORVALINE
DL-NORLEUCINE
L-NORLEUCINE
DL-LYSINE
L-LYSINE
N-ACETYL-L-METHIONINE
N-(3,5-DINITROBENZOYL)-DL-ALPHA-PHENYLGLYCINE
(S)-(+)-N-(3,5-DINITROBENZOYL)-ALPHA-PHENYLGLYCINE
N-(3,5-DINITROBENZOYL)-L-LEUCINE
L-ARGININE MONOHYDROCHLORII)E
L-2,4-DIAMINOBUTYRIC ACID I)IHYDROCHLORIDE
L-ORNITHINE MONOHYDROCHLOR7_DE
DL-LYSINE MONOHYDROCHLORIDF;
L-LYSINE MONOHYDROCHLORIDE
N-(TERT-BUTOXYCARBONYL)-L-~~SPARTIC ACID 4-BENZYL ESTER
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N-ALPHA-(TERT-BUTOXYCARBONYL)-L-GLUTAMINE
N-ALPHA-(TERT-BUTOXYCARBONYL)-L-HISTIDINE
BOC-LEU H20
BOC-L-METHIONINE
N-(TERT-BUTOXYCARBONYL)-L-TRYPTOPHAN
N-(TERT-BUTOXYCARBONYL)-L-VALINE
L-CYSTEINE HYDROCHLORIDE MONOHYDRATE
Z-L-TRYPTOPHAN
DL-TERT-LEUCINE
BOC-L-THREONINE
N-CBZ-L-THREONINE
N-CBZ-L-HISTIDINE
3,4-DEHYDRO-L-PROLINE
NALPHA-ACETYL-L-ASPARAGINE
L-LYSINE DIHYDROCHLORIDE
L-BUTHIONINE-(S, R)-SULFOXIMINE
P-AMINO-D-PHENYLALANINE
DL-BUTHIONINE (S, R)-SULFOXIMINE
3-CHLORO-L-ALANINE HYDROCHLORIDE
(S)-(-)-INDOLINE-2-CARBOXYLIC ACID
N-(PHOSPHONOMETHYL)GLYCINE, MONOISOPROPYLAMINE SALT
MIMOSINE
D-HOMOSERINE
(+)-OCTOPINE
N-CARBOBENZYLOXY-L-GLUTAMIC ACID 1-METHYL ESTER
DL-CYSTINE
3-(CARBOXYMETHYLAMINOMETHYL)-4-HYDROXYBENZOIC ACID
CIS-3-HYDROXY-DL-PROLINE
(S)-(-)-1,2,3,4-TETRAHYDRO-3-ISOQUINOLINECARBOXYLIC
ACID
N-(GAMMA-L-GLUTAMYL)-1-NAPHTHYLAMIDE MONOHYDRATE
3-(3,4-DIHYDROXYPHENYL)-2-METHYL-L-ALANINE,
SESQUIHYDRATE
DL-2-METHYLGLUTAMIC ACID HEMIHYDRATE
N-METHYL-D-ASPARTIC ACID MONOHYDRATE
DL-3-PHENYLSERINE HYDRATE
3-METHYL-L-HISTIDINE HYDRATE
N-{4-NITROBENZOYL)-L-GLUTAMIC ACID HEMIHYDRATE
4-NITRO-DL-PHENYLALANINE HYDRATE
L-CYSTEIC ACID MONOHYDRATE
4-AMINO-DL-PHENYLALANINE HYDRATE
D-ASPARAGINE MONOHYDRATE
D-LYSINE HYDRATE
L-(+)-CANAVANINE SULFATE MONOHYDRATE
D-CYSTEINE HYDROCHLORIDE MONOHYDRATE
D-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE
DL-PENICILLAMINE ACETONE ADDUCT HYDROCHLORIDE
MONOHYDRATE
2-METHYLORNITHINE HYDROCHLORIDE MONOHYDRATE
3-AMINO-L-TYROSINE DIHYDROCHLORIDE MONOHYDRATE
4-AMINO-L-PHENYLALANINE HYDROCHLORIDE HEMIHYDRATE
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L-GLUTAMIC ACID, MONOSODIUM SALT, MONOHYDRATE
3,5-DIIODO-L-TYROSINE DIHYDRATE
N-(9-FLUORENYLMETHOXYCARBONYL)-L-ALANINE MONOHYDRATE
3,5-DINITRO-L-TYROSINE MONOHYDRATE
4-NITRO-L-PHENYLALANINE MONOHYDRATE
DL-GLUTAMIC ACID MONOHYDRATE
L-CYSTEINESULFINIC ACID MONOHYDRATE
KAINIC ACID MONOHYDRATE
4-AMINO-L-PHENYLALANINE HYDRATE
IBOTENIC ACID MONOHYDRATE
L-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE
DL-HISTIDINE MONOHYDROCHLO:RIDE MONOHYDRATE
DL-CYSTEINE HYDROCHLORIDE MONOHYDRATE
DL-ARGININE HYDROCHLORIDE 7MONOHYDRATE
L-LYSINE MONOHYDRATE
L-ASPARAGINE MONOHYDRATE
DL-ASPARAGINE MONOHYDRATE
(S)-(-)-ALPHA-AMINOCYCLOHE:KANEPROPIONIC ACID HYDRATE
N-BENZYLGLYCINE HYDROCHLORIDE
2-AMINO-2-NORBORNANECARBOX'~'LIC ACID
CIS-4-AMINO-1-CYCLOHEXANECARBOXYLIC ACID
(+/-)-TRANS-3-AZABICYCLO(3.1.0)HEXANE-2-CARBOXYLIC ACID
3-AZETIDINECARBOXYLIC ACID
N-CARBOBENZYLOXY-L-TYROSINI~ HYDRATE
4-HYDROXY-2,2,6,6-TETRAMETHYL-4-PIPERIDINECARBOXYLIC
ACID,
and the like.
It is to be understood that the amino acids utilized
according to the present invention can also be selected
from proteiogenic amino acids or optical isomers thereof.
ATAs for use in the process of this invention for
preparation of macrocycle libraries are readily available
by synthesis from available amino alcohols, more
particularly protected-amino alcohol mesylates or
tosylates, and mercapto acids.
In the first step in preparation of ATA compounds of
the above formula wherein m = O, an aminothioether acid, a
compound of the formula
Q--CI -X
II
Rn Rs
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is reacted with about 2 to about 1.2 stoichiometric
equivalents of a mercaptide salt of a mercapto compound of
the formula
O
HS--W C-OH
wherein in the above formulas P, Rn, Rs, X and W are
defined above. The reaction is typically carried out in
solution phase, and a solution of the compound of Formula
II is added slowly to a solution of the mercaptide salt.
The mercaptide salt is most typically generated by
reacting the corresponding mercaptan in solution with
about two stoichiometric equivalent amounts of an alkali
metal alkoxide base, for example sodium methoxide. Other
mercaptide forming bases can be used (e. g. alkali metal
dimsylates or hydrides), but without advantage. Examples
of suitable mercapto acids for use in preparing the
library compounds of this invention include, but are not
intended to be limited to, the following:
THIOSALICYLIC ACID
N-ACETYL-DL-PENICILLAMINE
DL-PENICILLAMINE
2,3-DIMERCAPTOSUCCINIC ACID
MERCAPTOSUCCINIC ACID
N-(2-MERCAPTOPROPIONYL)GLYCINE
N-ACETYL-L-CYSTEINE
DL-CYSTEINE
3-MERCAPTOPROPIONIC ACID
DL-HOMOCYSTEINE
2-MERCAPTONICOTINIC ACID
D-CYSTEINE HYDROCHLORIDE
3-MERCAPTOBENZOIC ACID
4-MERCAPTOBENZOIC ACID
DL-2-MERCAPTOMETHYL-3-
GUANIDINOETHYLTHIOPROPANOIC ACID
2-THIOURACIL-5-CARBOXYLIC ACID
L-THIOHISTIDINE
4-MERCAPTOBUTYRIC ACID
CYS-GLY
DL-THIORPHAN
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4-MERCAPTOHYDROCINNAMIC' ACID
D-PENICILLAMINE
L-PENICILLAMINE
L-CYSTEINE
DL-CYSTEINE HYDROCHLORIDE
BOC-CYS-OH
GLUTATHIONE
D-CYSTEINE
MESO-ALPHA, ALPHA'-DIMERCAPTOADIPIC ACID
N-ACETYL-D-PENICILLAMINE
MERCAPTOTETRAZOLYLACETIC ACID
ALPHA-MERCAPTO-P-TOLUIC ACID
6-AZA-2-THIOURACIL-5-CARBOXYLIC ACID
2-THIOACETIC ACID-5-MERCAPTO-1,3,4-
THIADIAZOLE
N-ISOBUTYRYL-L-CYSTEINE
N-ISOBUTYRYL-D-CYSTEINE
CAPTOPRIL
5-MERCAPTO-TETRAZOLE-1-pROPIONIC ACID
SALOR S98,217-2
2,6-CSBA (2-Cl-6-merc~~ptobenzoic acid)
6-MERCAPTONICOTINIC ACI:~
MAYBRIDGE RJC 01025,
and the like.
In the preparation of aminothioether acid compounds
the mercaptide salt is reactE=d with a compound of the
Formula II. The substituent :~ is a good leaving group
subject to nucleophilic disp:Lacement by the mercaptide
salt. Exemplary of suitable X groups include, but are not
intended to be limited to, me~sylate, tosylate, halo, and
the like. Compounds of Formula II are typically derived
from an amino alcohol or protected-amino alcohol of the
formula
N 'Q CHOH
Rn Rs
wherein P, Rn, Q and RS are as defined above. The amino
alcohol is selected to have a molecular weight of about 60
to about 450, most typically about 60 to about 300. In
one preferred embodiment the amino alcohol starting
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material is reacted with methane sulfonyl chloride to
provide the corresponding mesylate in high yield using the
mesylation protocol of Crossland and Servis [JOC 35, 1952-
6 (1970)]. Suitable amino alcohols for use in preparing
the present library compounds include, but are not
intended to be limited to, the following:
TRAMS-2-AMINOCYCLOHEXANOL HYDROCHLORIDE
3-AMINOMETHYL-3,5,5-TRIMETHYLCYCLOHEXANOL
(1R,2R)-(-)-PSEUDOEPHEDRINE
L-ADRENALINE
DL-HOMOSERINE
Z-L-SERINE
2-ANILINOETHANOL
N-ACETYLETHANOLAMINE
2-(METHYLAMINO)ETHANOL
N-BENZYLETHANOLAMINE
2-(ETHYLAMINO)ETHANOL
DIETHANOLAMINE
2-(PROPYLAMINO)ETHANOL
D-SERINE
(1S,2S)-(+)-2-AMINO-1-PHENYL-1,3-PROPANEDIOL
D-ALLO-THREONINE
DIISOPROPANOLAMINE
2-AMINO-2-METHYL-1,3-PROPANEDIOL
TRIS(HYDROXYMETHYL)AMINOMETHANE
N-METHYL-D-GLUCAMINE
DL-2-AMINO-3-METHYL-1-BUTANOL
L-ISOLEUCINOL
L-PHENYLALANINOL
DL-4-CHLOROPHENYLALANINOL
L-METHIONINOL
CIS-4-HYDROXY-D-PROLINE
L-PROLINOL
3-PYRROLIDINOL
3-PYRROLIDINOL
2-PIPERIDINEMETHANOL
2-PIPERIDINEETHANOL
3-PIPERIDINEMETHANOL
4-HYDROXYPIPERIDINE
2-AMINO-2-METHYL-1-PROPANOL
D-(-)-ALPHA-PHENYLGLYCINOL
(-)-NOREPHEDRINE
DL-2-AMINO-1-PROPANOL
(+/-)-2-AMINO-1-BUTANOL
2-AMINO-1-PHENYLETHANOL
DL-ISOSERINE
1-AMINO-2-PROPANOL
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3-AMINO-1,2-PROPANEDIOL
DL-4-AMINO-3-HYDROXYBUTYRIC ACID
1,3-DIAMINO-2-PROPANOL
2-(2-AMINOETHYLAMINO)ETHANOL
ETHANOLAMINE
3-AMINO-1-PROPANOL
4-AMINO-1-BUTANOL
5-AMINO-1-PENTANOL
6-AMINO-2-HEXANOL
DL-2-AMINO-1-PENTANOL
DL-2-AMINO-1-HEXANOL
1-AMINO-1-CYCLOPENTANEMETHANOL
(1S,2S)-(+)-2-AMINO-3-METHOXY-1-PHENYL-1-PROPANOL
DL-PROPRANOLOL HYDROCHLORIDE
TRAMS-4-AMINOCYCLOHEXANOL HYDROCHLORIDE
TRIS(HYDROXYMETHYL)AMINOMETHANE HYDROCHLORIDE
DL-SERINE METHYL ESTER HYDROCHLORIDE
L-SERINE ETHYL ESTER HYDROCHLORIDE
L-PHENYLEPHRINE HYDROCHLORIDE
3-HYDROXYPIPERIDINE HYDF;OCHLORIDE
DL-OCTOPAMINE HYDROCHLOF;IDE
DL-NORMETANEPHRINE HYDRC)CHLORIDE
ETHANOLAMINE HYDROCHLORIDE
3-HYDROXYPIPERIDINE
N-CYCLOHEXYLETHANOLAMINE
L -NOF;ADRENAL INE
L -ADRENAL INE B I TARTFLF,TE
D-SPHINGOSINE
N-(TERT-BUTOXYCARBONYL)-L-SERINE
N-ACETYL-DL-SERINE
L-THREONINE METHYL ESTER HYDROCHLORIDE
L-ARGININIC ACID
D-GLUCOSAMINIC ACID
L-TYROSINOL HYDROCHLORIDE
L-SERINE BENZYL ESTER HYDROCHLORIDE
2-AMINO-2-METHYL-1-PROPAI~OL HYDROCHLORIDE
2-AMINO-1,3-PROPANEDIOL OXALATE
METHYL 3-AMINO-3-DEOXY-BETA-D-GLUCOPYRANOSIDE
2-AMINO-1,3-PROPANEDIOL
DL-SERINE HYDROXAMATE
L-SERINE BETA-NAPHTHYLAM:IDE
N-CBZ-D-GLUCOSAMINE
N-(TERT-BUTOXYCARBONYL)E'L'HANOLAMINE
DL-THREONINE HYDROXAMATE
SER-BETA-ALA
N-T-BOC-L-HOMOSERINE
METHYL 3-AMINO-3-DEOXY-ALPHA-D-MANNOPYRANOSIDE
HYDROCHLORIDE
L-THREONINAMIDE HYDROCHLORIDE
3-AMINO-2,2-DIMETHYL-1-PF;OPANOL
L-HOMOSERINE
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N-T-BOC-D-SERINE
N-CARBOBENZOXY-DL-SERINE
L-LEUCINOL
L-SERINE METHYL ESTER HYDROCHLORIDE
DL-THREONINE
DL-SERINE
L-SERINE
(+)-PSEUDOEPHEDRINE
(-)-EPHEDRINE
L-ALLO-THREONINE
D-THREONINE
L-THREONINE
L-VALINOL
D-VALINOL
D-PHENYLALANINOL
CIS-4-HYDROXY-L-PROLINE
L-HYDROXYPROLINE
D-PROLINOL
L-(+)-ALPHA-PHENYLGLYCINOL
(1S,2R)-(+)-PHENYL-PROPANOLAMINE
(S)-(+)-2-AMINO-1-PROPANOL
D-ALANINOL
(S)-(+)-2-AMINO-1-BUTANOL
(R)-(-)-2-AMINO-1-BUTANOL
(R)-(-)-1-AMINO-2-PROPANOL
(S)-(+)-1-AMINO-2-PROPANOL
(+)-EPHEDRINE HYDROCHLORIDE
D-MANNOSAMINE HYDROCHLORIDE
D(+)-NOREPHEDRINE HYDROCHLORIDE
NOREPHEDRINE HYDROCHLORIDE
BOC-L-THREONINE
N-CBZ-L-THREONINE
DL-ALLOTHREONINE
L-SERINAMIDE HYDROCHLORIDE
ALPHA-D-GLUCOSAMINE HYDROCHLORIDE
DL-METHIONINOL
(1S,2R)-(+)-2-AMINO-1,2-DIPHENYLETHANOL
(1R,2S)-(-)-2-AMINO-1,2-DIPHENYLETHANOL
S-BENZYL-L-CYSTEINOL
BOC-L-PHENYLALANINOL
L-HISTIDINOL DIHYDROCHLORIDE
D-(-)-THREO-2-AMINO-1-(4-NITROPHENYL)-1,3-
PROPANEDIOL
ETHAMBUTOL DIHYDROCHLORIDE
(+/-}-ARTERENOL BITARTRATE SALT
S-ALPHA-HYDROXYMETHYL TYROSINE
(1S,2S)-(+)-THIOMICAMINE
CIS-2-AMINOMETHYL-1-CYCLOHEXANOL HYDROCHLORIDE
CIS-2-HYDROXYMETHYL-1-CYCLOHEXYLAMINE HYDROCHLORIDE
TRANS-2-AMINOMETHYL-1-CYCLOHEXANOL
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TRAMS-2-HYDROXYMETHYL-1-CYCLOHEXYLAMINE
HYDROCHLORIDE
DL-3-PHENYLSERINE HYDRATE
N-(2-HYDROXYETHYL)CARBAMIC ACID BENZYL ESTER
N-(TERT-BUTOXYCARBONYL)-L-SERINE METHYL ESTER
(R)-(+)-3-HYDROXYPIPERIDINE HYDROCHLORIDE
(S)-TERT-LEUCINOL
N-TRITYL-L-SERINE METHYL ESTER
3-PHENYL-DL-SERINE
3-PHENYL-DL-SERINE
(+/-)-NOREPINEPHRINE L-:BITARTRATE HYDRATE
(1S,2R)-(-)-CIS-1-AMINO-2-INDANOL
4-AMINO-2-BUTANOL
2-[2-(AMINOPHENYLTHIO]B:ENZYL ALCOHOL,
and the like.
Amino alcohol starting materials can also be prepared
in high yield from available amino acids and protected
amino acids of the formula
N Q' COOH
III
Rn
wherein P, Rn and Q are as defined above. Available amino
acid compounds can be protected with an amino-protecting
group and converted in good ~~ields to the corresponding
protected-amino alcohols of t:he formula
CHOH
Rn Rs
For example, when Rs is hydrogen these compounds can be
obtained from sodium borohydride reduction of the mixed
carbonate formed, for example, between the starting acid
and isobutyryl chloroformate. Any of the above
exemplified amino acids are available for conversion to
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the corresponding amino alcohols for use in preparing
amino thioether acids for use in this invention.
Where the thioether-forming reaction is carried out
using a dialkali metal of a mercaptoacid in excess of a
stoichiometric amount relative to the amino alcohol
derived compound of Formula II above, the excess reagent
frequently migrates with the thioether acid product on a
flash silica column and cannot be removed by simple
extraction. The excess acid can be separated from the
product by dissolving the reaction mixture in ethyl
acetate and adding a 2-fold molar excess of solid mercuric
acetate to the solution. After 3-20 hours the resulting
slurry is filtered through a celite pad, the filtrate
containing the desired thioether product is subjected to
flash chromatography on silica, and the chromatographed
material is evaporated to provide the purified reaction
product.
Reaction Scheme II below (bold-face numberals
correspond to compound numerals in Scheme II) illustrates
of the preparation of ATA compounds, for use in accordance
with this invention using amino acid starting materials,
such as, for example, amino acid 1 [Formula III, where P =
Rn = hydrogen and Q =
C
R1 R2
wherein R1 is hydrogen and R2 - CH2R]. With reference to
Scheme II below, amino acid 1 is protected, reduced to the
corresponding alcohol, and then converted to mesylate 2.
The ATA is formed by condensing the disodium salt of a
mercapto acid with mesylate 2 to form 3a. The next step
of the process involves exchanging protecting groups to
yield a N-Fmoc-protected ATA 3b. Each of the reaction
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steps proceed smoothly and c;~n be carried out on multigram
scale.
R O R OMs S ' W OOOH
R S-- W~-COOH
~ a, b, c
'OH ->
- a
NH2 NHP NHP
3b
3a P=Fmoc
P=Alloc P=Alloc
or Boc or Boc
Scheme II
a) Allyl chloroformate or d=-t-butyl dicarbonate, Na2C03,
1:1 H20:dioxane, 4oC, 90-950, P=Alloc or Boc,
respectively;
b) isobutyryl chloroformate, N-methylmorpholine (NMM) in
ethylene glycol dimethy:l ehter (DME), -lSoC, then aq
NaBH4, -lSoC, 65-90a;
c) CH3S02C1, TEA in THF, 4oC, 90-95%;
d) mercapto acid (1eq), NaOMe (2eq) in DMF, r.t., 65-950;
e) P= Alloc~Fmoc, PPh3 (0.25eq), Pd(PPh3)4 (0.03eq), HOAc
(2eq) in THF, then Fmc>c-N-hydroxysuccinimide ester,
Na2C03 in 1:1 H20:dio:~ane, 4oC to r.t., 500; P=
Boc-~Fmoc, 4N HCl in clioxane or EtOAc, 1.5h, then
Na2C03 and Fmoc-OSu in 1:1 H20:dioxane, 90-950.
The aminothioether compounds can be converted to
their corresponding sulfoxides (m=1) and sulfones (m=2)
(included in the ATAs useful in accordance with this
invention) by oxidation. The oxidation can be
accomplished in high yields using stoichiometric amounts
of any one of several oxidizing agents including m-
chloroperbenzoic acid, period,~te, and oxone (a mixture of
potassium hydrogen persulfate, potassium bisulfate and
potassium sulfate). The oxidation reaction is carried out
in any one of a wide variety of solvents at a temperature
of about -10 C to about 30°C. The preparation of
sulfoxides and sulfones by oxidation of thioethers are
well known in the art.
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Oxidation of a sulfide to its corresponding sulfoxide~
1). P. G. Hunt, J. I. Grayson, S. Warren and J. Durman _J.
Chem. Soc. Perkin Trans 1 (1986), p. 1939. [Uses m-
CPBA (meta-chloro-peroxybenzoic acid) or sodium periodate
to give desired products in 92-1000 yield].
2). D. M. Hedstrand, S. R. Byrn, A. T. McKenzie and P. L.
Fuchs J. Ora. Chem. 52 (1987), pp. 592-598.
3). C. Jouen, M. C. Lasne and J. C. Pommelet Tet. Lett.
37 (1996), pp. 2413-2416.
Oxidation of a sulfide to its corres~ondina sulfone~
4). B. M. Trost and R. Braslau J. Ora. Chem. 53 (1988),
pp. 532-537. [Uses Oxone from DuPont which is "...a
mixture of potassium hydrogen persulfate, potassium
bisulfate and potassium sulfate...."].
Amino-protected ATAs useful in the present process
have a molecular weight of about 150 to about 800. A
preferred amino-protecting group for the amino acids and
ATAs used in this process is 9-fluorenyloxymethylcarbonyl.
According to the process of the present invention,
following the first amino acid or ATA coupling reaction of
process step (c) as described above, in process step (d)
the amino-protecting group is removed from the solid
support coupled product following the same procedures as
described above for process step (b). Thereafter, process
step (c) and process step (d) may be repeated one or more
times using the same or a different amino-protected amino
acid or amino-protected ATA. However, at least one of the
process steps (c) and (e) is carried out using an amino-
protected ATA.
In the next process step (f) for preparing the
present library compounds, the support-bound product
resulting from process step (d) (or process step (e), if
optionally performed) is coupled with an organic acid of
the formula
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1
L --COON
2I
X
where L is as defined above and X is a leaving group
subject to nucleophilic displacement. Exemplary of such
acids include, but are not intended to be limited to:
bromoacetic acid
alpha-bromophenylacetic acid
alpha-bromopropionic acid
iodoacetic acid
chloroacetic acid
4-carboxybenzyl bromide
4-carboxymethylbenzyl bromide
3-carboxybenzyl bromide
2-carboxybenzyl bromide,
and the like.
The coupling of the organic <~cid with the terminal amino
group on the solid support bound product is carried out
under essentially the same rE~action conditions and
stoichiometry described above for the coupling of the
protected amino acids or the protected ATAs in steps (c)
and (e) above except that the hydroxybenzotriazole (HOBT)
reagent typically used with <:arbodiimides to form the
intermediate "active" HOBT e:~ter is not used in the
coupling reaction mixture due to the fact that HOBT is
sufficiently nucleophilic that it reacts to displace the
terminal leaving group. The coupling reaction proceeds
without added HOBT via the active ester formed by the
reaction of the organic acid and the carbodiimide
reactant.
In the next step of the process for the preparation
.. of the present macrocycle library compounds, the support- .
bound product from step (f) above is cyclized and cleaved
from the solid support. While those two reactions can be
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conducted in either order, i.e., cyclization before
cleavage, or cleavage before cyclization, it is preferred
that cyclization be conducted in solution phase after
cleavage of the acyclic precursor from the solid support.
The optimum reaction conditions for cleavage of the
product of step (f) or of the pre-cyclized product from
the solid support are dependent on the nature of the acid
reactive groups on the support. Typically such cleavage
reactions are carried out by treating the solid support
with trifluoroacetic acid in the presence of water,
optionally in the presence of triethylsilane, conditions
which also effect removal of the thio-protecting group
from the acyclic precursor product. The cleavage product
is isolated by separation of the "cleavage cocktail"
25 solution from the solid support and lyophilization of the
"cleavage cocktail". Cyclization of the acyclic
macrocycle precursor product can be efficiently carried
out in solution in the presence of a non-nucleophilic base
such as, for example, 2,6-lutidine, diisopropylethylamine
(DIPEA), and the like.
When the acid reactive group on the solid support
comprises a covalently bound group of the formula
O
HN T N CO-CH2-C6H~0-CH2-CsHd-Polymer
Ry Ry
the product macrocycle library compounds of the invention
are those of Formula I wherein B is a group of the formula
--- N T NH
Ry Ry ~,
wherein Ry, T and Ry~~ are as defined above. Such library
compounds are optionally reacted with an electrophilic
agent having a molecular weight of about 30 to about 600
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to provide library compounds of Formula I wherein E is a
substituent derived from an electrophilic reagent.
Suitable electrophilic ageni:s include, but are not
intended to be limited to, organic halides, acyl halides,
sulfonic acid esters, organohaloformates, organosulfonyl
halides, organic isocyanate;~, organic isothiocyanates,
aldehydes, ketones, and the like. Examples of such
electrophilic agents include, but are not intended to be
limited to:
3,5-bis(trifluoromethy7.)benzoyl chloride
benzoyl chloride
2-bromobenzoyl chloride
2-fluorobenzoyl chloride
pentafluorobenzoyl chloride
2,4-difiuorobenzoyl chloride
2,6-difluorobenzoyl chloride
2-chlorobenzoyl chloride
2,4-dichlorobenzoyl chloride
2,6-dichlorobenzoyl chloride
o-acetylsalicyloyl chloride
2-methoxybenzoyl chloride
2,6-dimethoxybenzoyl chloride
2-(trifluoromethyl)benzoyl chloride
o-toluoyl chloride
3-bromobenzoyl chloride
3-fluorobenzoyl chloride
3-chlorobenzoyl chloride
3,4-dichlorobenzoyl chloride
m-anisoyl chloride
3,4-dimethoxybenzoyl chloride
3,4,5-trimethoxybenzoyl chloride
3,5-dimethoxybenzoyl chloride
3-ethoxybenzoyl chloride
isophthaloyl chloride
trimesoyl chloride
3-(trifluoromethyl)benzoyl chloride
m-toluoyl chloride
3-(chloromethyl) benzoyl chloride
4-bromobenzoyl chloride
4-fluorobenzoyl chloride
4-chlorobenzoyl chloride
p-anisoyl-chloride
4-ethoxybenzoyl chloride
4-n-butoxybenzoyl chloride
4-n-hexyloxybenzoyl chl~~ride
4-heptyloxybenzoyl chloride
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4-biphenylcarbonyl chloride
terephthaloyl chloride
4-(trifluoromethyl)benzoyl chloride
4-tert-butylbenzoyl chloride
p-toluoyl chloride
4-ethylbenzoyl chloride
4-n-propylbenzoyl chloride
4-butylbenzoyl chloride
4-pentylbenzoyl chloride
4-hexylbenzoyl chloride
4-n-heptylbenzoyl chloride
methyl oxalyl chloride
ethyl oxalyl chloride
heptafluorobutyryl chloride
2-acetoxyisobutyryl chloride
pivaloyl chloride
3-chloropivaloyl chloride
2-bromopropionyl chloride
2,3-dibromopropionyl chloride
2,3-dichloropropionyl chloride
o-acetylmandelic acid chloride
itaconyl chloride
methacryloyl chloride
isobutyryl chloride
2-ethylhexanoyl chloride
acetyl chloride
bromoacetyl chloride
chloroacetyl chloride
phenoxyacetyl chloride
4-chlorophenoxyacetyl chloride
methoxyacetyl chloride
phenylacetyl chloride
3,3-dimethylacryloyl chloride
cinnamoyl chloride
fumaryl chloride
ethyl malonyl chloride
tert-butylacetyl chloride
isovaleryl chloride
undecanoyl chloride
lauroyl chloride
myristoyl chloride
palmitoyl chloride
heptadecanoyl chloride
stearoyl chloride
propionyl chloride
3-bromopropionyl chloride
3-chloropropionyl chloride
hydrocinnamoyl chloride
succinyl chloride
3-carbomethoxypropionyl chloride
ethyl succinyl chloride
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butyryl chloride
4-bromobutyryl chloride
4-chlorobutyryl chloride
valeryl chloride
5-chlorovaleryl chloride
adipoyl chloride
hexanoyl chloride
6-bromohexanoyl chloride
pimeloyl chloride
heptanoyl chloride
suberoyl chloride
octanoyl chloride
10-undecenoyl chloride
2-chloro-2,2-diphenylac~etyl chloride
dichloroacetyl chloride
alpha-chlorophenylacet5rl chloride
2-chloropropionyl chloride
2-iodobenzoyl chloride
4-iodobenzoyl chloride
cyclopropanecarbonyl chloride
traps-2-phenyl-1-cyclopropanecarbonyl chloride
cyclobutanecarbonyl ch~_oride
cyclopentanecarbonyl chloride
3-cyclopentylpropionyl chloride
cyclohexanecarbonyl chloride
4-cyanobenzoyl chloride
2-furoyl chloride
1-naphthoyl chloride
2-naphthoyl chloride
thiophene-2-carbonyl chloride
2-thiopheneacetyl chloride
trimellitic anhydride chloride
2,6-pyridinedicarboxylic acid chloride
2-quinoxaloyl chloride
2-nitrobenzoyl chloride
3-nitrobenzoyl chloride
3,5-dinitrobenzoyl chloride
4-nitrobenzoyl chloride
3,4-dimethoxyphenylacetyl chloride
3-methyladipoyl chloride
3,5-dichlorobenzoyl chloride
2,5-difluorobenzoyl chloride
3,4-difluorobenzoyl chloride
9-fluorenone-4-carbonyl chloride
3,5-difluorobenzoyl chloride
(s)-(-)-n-(trifluoroacetyl)prolyl chloride
benzyloxyacetyl chloride
acetoxy acetyl chloride
3-cyanobenzoyl chloride
2,5-dimethoxyphenylacet~~l chloride
3-methoxyphenylacetyl chloride
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iminodibenzyl-5-carbonyl chloride
2,4,6-trimethylbenzoyl chloride
tetrafluorosuccinyl chloride
perfluorooctanoyl chloride
diphenylacetyl chloride
alpha-methyl valeroyl chloride
methyl malonyl chloride
ethyl glutaryl chloride
5-bromovaleryl chloride
methyl adipyl chloride
3-cyclohexenecarbonyl chloride
3-isocyanato benzoyl chloride
2,4,6-triisopropylbenzoyl chloride
fluoroacetyl chloride
2-ethoxybenzoyl chloride
piperonyloyl chloride
2,4-dimethoxybenzoyl chloride
2,3,5,6-tetrachloroterephthaloyl chloride
5-(dimethylsulfamoyl)-2-methoxybenzoyl chloride
2-(4-chlorobenzoyl)benzoyl chloride
2,2-bis(chloromethyl)propionyl chloride
cinnamylidenemalonyl chloride
2-phenoxypropionyl chloride
2-phenylbutyryl chloride
2-ethylbutyryl chloride
p-tolylacetyl chloride
gamma-methylvaleroyl chloride
3,3-dichloropivaloyl chloride
1-methyl-1-cyclohexanecarboxylic acid chloride
2-(2,4,5-trichlorophenoxy)acetyl chloride
4-chloro-3-nitrobenzoyl chloride
4-methyl-3-nitrobenzoyl chloride
2,3-dichlorobenzoyl chloride
morpholine-4-carbonyl chloride
p-chlorophenylacetyl chloride
bicyclo[2.2.1]heptane-2-carbonyl chloride
d(-)-alpha-formyloxy-alpha-phenylacetyl chloride
d(-)-alpha-phenylglycine chloride hydrochloride
trifluoroacetyl chloride
pentafluoropropionyl chloride
hexafluoroglutaryl chloride
2-chlorocinnamoyl chloride
o-methoxycinnamyl chloride
5-nitro-2-furoyl chloride
2-chlorobutyryl chloride
4-phenylazobenzoyl chloride
4-n-amyloxybenzoyl chloride
4-decylbenzoyl chloride
4-octylbenzoyl chloride
dl-2-methylbutyryl chloride
linolenoyl chloride
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linolelaidoyl chloride
llh-eicosafluoroundecanoyl chloride
9h-hexadecafluorononanoyl chloride
2,3-difluorobenzoyl chloride
2-(benzoyloxymethyl)benzoyl chloride
2,2-dimethylvaleroyl chloride
3,5,5-trimethylhexanoyl chloride
phenothiazine-10-carbonyl chloride
3,4-dimethyl benzoyl chloride
(+)-p-(2-methylbutyl)benzoyl chloride
2,4-dichlorophenoxyacetic chloride
pentadecanoyl chloride
nonadecanoyl chloride
neoheptanoyl chloride
9-anthracenecarbonyl chloride
2-ethoxy-1-naphthoyl chloride
pyrrolidine carbonyl chloride
m-(chlorosulfonyl)benzoyl chloride
2-n-propyl-n-valeroyl chloride
2-chloro-4-nitrobenzoyl chloride
2-phenoxybutyryl chloride
2-chloronicotinyl chloride
6-chloronicotinyl chloride
4-(trifluoromethoxy)benzoyl chloride
2-(trifluoromethoxy)benzoyl chloride
2,6-dichloropyridine-4-carbonyl chloride
3-chlorobenzo(blthiophene-2-carbonyl chloride
4-chloromethylbenzoyl chloride
neodecanoyl chloride
(phenylthio)acetyl chloride
4-carbethoxyhexafluorobutyryl chloride
octafluoroadipoyl chloride
2-diazo-3,3,3-trifluoro;r~ropionylchloride
2-bromobutyryl chloride
arachidoyl chloride
cis-vaccenoyl chloride _
11-eicosenoyl chloride
behenoyl chloride
petroselinoyl chloride
palmitoleoyl chloride
tridecanoyl chloride
2-chloro-5-nitrobenzoyl chloride
3-methylthiopropionyl chloride
methyl 4-chlorocarbonylbenzoate
anthraquinone-2-carbony:L chloride
carbazole-n-carbonyl ch:Loride
2-nitrophenoxyacetyl ch:Loride
2-bromo-2-methylpropionyl chloride
2-fluoro-3-(trifluoromet:hyl)benzoyl chloride
2-fluoro-4-(trifluoromet:hyl)benzoyl chloride
2-fluoro-5-(trifluoromet:hyl)benzoyl chloride
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3-fluoro-5-(trifluoromethyl)benzoyl chloride
4-fluoro-2-(trifluoromethyl)benzoyl chloride
4-fluoro-3-(trifluoromethyl)benzoyl chloride
2-fluoro-6-(trifluoromethyl)benzoyl chloride
2,3,6-trifluorobenzoyl chloride
2,4,5-trifluorobenzoyl chloride
2,4-di(trifluoromethyl)benzoyl chloride
2,6-di(trifiuoromethyl)benzoyl chloride
3-(trifluoromethoxy)benzoyl chloride
m-(fluorosulfonyl)benzoyl chloride
trans-1,2-cyclobutanedicarboxylic acid chloride
3-cyclohexylpropionyl chloride
4-ethyl-2,3-dioxo-1-piperazinecarbonylchloride
isoxazole-5-carbonyl chloride
bromodifluoroacetyl chloride
erucoyl chloride
2,4,6-trifluorobenzoyl chloride
dichlorochrysanthemic acid chloride
isononanoyl chloride
1-adamantanecarbonyl chloride
2,5-bis(trifluoromethyl)benzoyl chloride
2,3,4-trifluorobenzoyl chloride
2,3,4,5-tetrafluorobenzoyl chloride
2,4,6-trichlorobenzoyl chloride
2,4-dichloro-5-fluorobenzoyl chloride
4-methoxyphenylacetyl chloride
trans-3-(trifluoromethyl)cinnamoyl chloride
3-(dichloromethyl) benzoyl chloride
4-isocyanato benzoyl chloride
heneicosanoyl chloride
2-chloroisobutyryl chloride
trans-4-nitrocinnamoyl chloride
3,4,5-trifluorobenzoyl chloride
5-fluoro-2-(trifluoromethyl)benzoyl chloride
2,3,5-trifluorobenzoyl chloride
2-chloro-4-fluorobenzoyl chloride
(-)-alpha-chlorophenylacetyl chloride
2-(para-tolylsulfonyl)acetyl chloride
4-methyl-4-nitrohexanoyl chloride
1-chloro-4-fluorosulfonyl-2-naphthoyl chloride
2,3-dibromo-3-phenylpropionyl chloride
2-menthoxyacetyl chloride
2-phenyl-2-(phenylsulfonyl)acetyl chloride
4,4,4-trifluorocrotonyl chloride
4,4,4-trifluorobutyryl chloride
3,4-dichloro-2,5-thiophenedicarbonyl chloride
pentachlorobenzoyl chloride
4,4,7,7-tetranitrosebacoyl chloride
alpha,alphal-dimethylsuccinyl chloride
alpha-bromoisovaleryl chloride
benzoyl chloride
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_. 5 ~ -
oleoyl chloride
methyl suberyl chloride
gamma-linolenoyl chloride
(-)-camphanic acid chloride
S 4,4'-stilbenedicarbony7_ chloride
chlorinated benzoyl ch~.oride
(1r)-(+)-camphanic chloride
2-(4-nitrophenoxy)tetradecanoyl chloride
7-[(chlorocarbonyl)methoxy]-4-methylcoumarin
n,n-bis(2-chloroethyl)carbamoyl chloride
(s)-(-)-2-acetoxypropionyl chloride
linoleoyl chloride
3-chlorotetrafluoroprop~ionyl chloride
3,4-dichloropentafluorobutyryl chloride
7h-dodecafluoroheptanoyl chloride
5h-octafluoropentanoyl chloride
perfluorononanoyl chloride
3h-tetrafluoropropionyl chloride
2-bromo-2,3,3,3-tetrafluoropropanoyl chloride
arachidonoyl chloride
pentachloropropionyl chloride
4-decenoyl chloride
tridecafluoroheptanoyl chloride
undecafluorocyclohexanecarbonyl chloride
4-n-nonylbenzoyl chloride
3-(trichlorogermyl)propionylchloride
3,4,5-triiodobenzoyl chloride
2-(phenylthio)propionyl chloride
2,2,2-triphenylacetyl chloride
d(-)-alpha-azido-phenyl acetyl chloride
4-azido-benzoyl chloride
difluoroacetyl chloride
5-chloropyrazine-2-carbonyl chloride
n-(1-naphthalenesulfony.L)-1-phenylalanyl chloride
n-(4-nitrophenylsulfony:L)-1-phenylalanyl chloride
n-(p-toluenesulfonyl)-1-phenylalanyl chloride
dimethylmalonyl chloride
methyl sebacoyl chloride
2,5-:dichloropyridine-3~-carbonyl chloride
3-(2,5 xylyloxy) propionyl chloride,
and the like.
Organic Halides:
benzyl bromide
alpha-bromo-o-xylene
alpha-bromo-m-xylene
4-(tert-butyl)benzyl bromide
alpha-bromo-p-xylene
tert-butyl bromoacetate
methyl bromoacetate
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benzyl bromoacetate
ethyl bromoacetate
2-bromoacetophenone
2-bromo-2'-methoxyacetophenone
2-bromo-2',4'-dimethoxyacetophenone
2-bromo-2',5'-dimethoxyacetophenone
3-methoxyphenacyl bromide
2-bromo-4'-methoxyacetophenone
2-bromo-4'-phenylacetophenone
2-bromo-4'-methylacetophenone
ethyl bromopyruvate
1-bromopinacolone
1-bromo-2-butanone
1-bromo-2,2-dimethoxypropane
1-bromo-2,2-dimethylpropane
bromoacetaldehyde dimethyl acetal
bromoacetaldehyde diethyl acetal
1-bromo-2-methylpropane
1-bromo-2-ethylbutane
2-ethylhexyl bromide
I-bromodecane
1-bromoundecane
2-bromoacetamide
iodoacetamide
4-(bromomethyl)phenylacetic acid phenacyl ester
isopropyl bromoacetate
5-bromo-2-methyl-2-pentene
3,4-difluorobenzyl bromide
2,5-difluorobenzyl bromide
3,5-bis(trifluoromethyl)benzyl bromide
2-bromo-2'-nitroacetophenone
3,5-difluorobenzyl bromide
2,4-bis(trifluoromethyl)benzyl bromide
8-bromo-1-octanol
4-(bromomethyl)phenylacetic acid
methyl (r)-(+)-3-bromo-2-methylpropionate
4-iodobutyl acetate
7-acetoxy-4-bromomethylcoumarin
4-bromomethyl-6,7-dimethoxycoumarin
2,4-difluorobenzyl bromide
methyl 2-(bromomethyl)acrylate
3-bromopropionaldehyde dimethyl acetal.
(r)-(-)-3-bromo-2-methyl-1-propanol,
and the like.
Sulfonic Acid Esters:
ethyl trifluoromethanesulfonate
2,2,2-trifluoroethyl p-toluenesulfonate
2-chloroethyl-p-toluenesulfonate
1,3-propane sultone
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_.5g_
5'-tosyladenosine
1,4-butane sultone
cyanomethyl benzenesulf:onate
hexadecyl methanesulfonate
ethyl methanesulfonate
2-chloroethyl methanesulfonate
ethyl p-toluenesulfonat.e
trans-2-hydroxycyclohexyl p-toluenesulfonate
(2r)-(-)-glycidyl tosyl.ate
(s)-(+)-2-methylbutyl methanesulfonate
(s)-(+)-2-methylbutyl ~~-toluenesulfonate
(s)-(+)-1-phenyl-1,2-ethanediol 2-tosylate
(2r)-(-)-glycidyl 3-nitrobenzenesulfonate
propargyl benzenesulfon.ate
2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-
toluenesulfonate
(r)-(-)-2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-
toluenesulfonate
(s)-(+)-2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-
toluenesulfonate
1,2:5,6-di-o-isopropylidene-3-o-(methylsulfonyl)-
alpha-
d-glucofuranose
ethyl 1-2-((methylsulfonyl)oxy)propionate
(2s)-(+)-glycidyl tosylate
(2s)-(+)-glycidyl 3-nitrobenzenesulfonate
3-o-acetyl-6-o-benzoyl-5-o-(methylsulfonyl)-1,2-0-
isopropylidene-alpha-d-glucofu
(r)-(-)-1-benzyloxy-3-(p-tosyloxy)-2-propanol
(s)-(+)-1-benzyloxy-3-(p-tosyloxy)-2-propanol
ethyl 1-2-((trifluoromethylsulfonyl)oxy)propionate
2-(2-chloroethoxy)ethyl methanesulfonate
1-cyanoethyl p-toluenesulfonate,
and the like.
Organohaloformates:
9-fluorenylmethyl chlor~~formate
phenyl chloroformate
4-chlorophenyl chlorofo:rmate
methyl chloroformate
benzyl chloroformate
vinyl chloroformate
isobutyl chloroformate
2-ethylhexyl chloroform<~te
ethyl chloroformate
2-bromoethyl chloroformate
2-chloroethyl chloroformate
1-chloroethyl chloroformate
allyl chloroformate
n-propyl chloroformate
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butyl chloroformate
n-hexyl chloroformate
octyl chloroformate
2,2,2-trichloro-1,1-dimethylethyl chloroformate
2,2,2-trichloroethyl chloroformate
cholesteryl chloroformate
4-nitrophenyl chloroformate
4-nitrobenzyl chloroformate
(-)-menthyl chloroformate
4-t-butylcyclohexyl chloroformate
cetyl chloroformate
(+)-1-(9-fluorenyl)ethyl chloroformate
isopropyl chloroformate
3-chlorocyclohexyl chloroformate
decyl chloroformate
oleyl chloroformate
octadecyl chloroformate
butenediol bischloroformate
2-chlorobenzyl chloroformate
4-chlorobutyl chloroformate
(+)-menthyl chloroformate
4,5-dimethoxy-2-nitrobenzyl chloroformate
cyclopentyl chloroformate
t-butylcyclohexyl chloroformate
menthylchloroformate
p-tolyl chloroformate
4-bromophenyl chloroformate
4-fluorophenyl chloroformate
4-methoxyphenyl chloroformate
2-nitrophenyl chloroformate
4-methoxycarbonylphenyl chloroformate
1-chloro-2-methylpropyl chloroformate
(+/-)-1,2,2,2-tetrachloroethyl chloroformate
2,2-dichloroethyl chloroformate
myristyl chloroformate
cyclohexyl chloroformate
chloromethyl chloroformate,
and the like.
Organosulfonylhalides:
1-naphthalenesulfonyl chloride
dansyl chloride
2-naphthalenesulfonyl chloride
2-acetamido-4-methyl-5-thiazolesulfonyl chloride
2-thiophenesulfonyl chloride
B-quinolinesulfonyl chloride
benzenesulfonyl chloride
pentafluorobenzenesulfonyl chloride
2,5-dichlorobenzenesulfonyl chloride
2-nitroberizenesulfonyl chloride
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2,4-dinitiobenzenesulfonyl chloride
3,5-dichloro-2-hydroxybenzenesulfonyl chloride
2,4 6-triisopropylbenzE~nesulfonyl chloride
2-mesitylenesulfonyl chloride
3-nitrobenzenesulfonyl chloride
p-bromobenzenesulfonyl chloride
4-fluorobenzenesulfonyl. chloride
4-chlorobenzenesulfonyl. chloride
4-chloro-3-nitrobenzene~sulfonyl chloride
20 pipsyl chloride
4-nitrobenzenesulfonyl chloride
4-methoxybenzenesulfonyl chloride
4-tert-butylbenzenesulfonyl chloride
p-toluenesulfonyl chloride
trifluoromethanesulfonyl chloride
trichloromethanesulfonyl chloride
isopropylsulfonyl chloride
methanesulfonyl chloride
alpha-toluenesulfonyl chloride
trans-beta-styrenesulfonyl chloride
2,2,2-trifluoroethanesulfonyl chloride
1-hexadecanesulfonyl chloride
ethanesulfonyl chloride
2-chloroethanesulfonyl chloride
1-propanesulfonyl chloride
3-chloropropanesulfonyl chloride
1-butanesulfonyl chloride
methyl 2-(chlorosulfony;L)benzoate
2-vitro-4-(trifluoromethyl)benzenesulfonyl chloride
3-(trifluoromethyl)benzenesulfonyl chloride
1-octanesulfonyl chloride
4-(trifluoromethoxy)benzenesulphonyl chloride
(1r)-(-)-10-camphorsulfonyl chloride
d-(+)-10-camphorsulfonyl. chloride
(+/-)-10-camphorsulfonyl. chloride
2-vitro-alpha-toluenesul.fonyl chloride,
and the like.
Isocyanate Reagents:
trans-2-phenylcyclopropyl isocyanate
phenyl isocyanate
2-bromophenyl isocyanate
2-fluorophenyl isocyanate
2,4-difluorophenyl isocyanate
2,6-difluorophenyl isocyanate
2-chlorophenyl isocyanate
2,3-dichlorophenyl isocy.~nate
2,4-dichlorophenyl isocy,~nate
2,5-dichlorophenyl isocy~nate
2,6-dichlorophenyl isocy~~nate
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2-methoxyphenyl isocyanate
2,4-dimethoxyphenyl isocyanate
2,5-dimethoxyphenyl isocyanate
2-ethoxyphenyl isocyanate
2-(trifluoromethyl)phenyl isocyanate
o-tolyl isocyanate
2,6-dimethylphenyl isocyanate
2-ethylphenyl isocyanate
3-bromophenyl isocyanate
3-fluorophenyl isocyanate
3-chlorophenyl isocyanate
3,4-dichlorophenyl isocyanate
3-methoxyphenyl isocyanate
3-(trifluoromethyl)phenyl isocyanate
m-tolyl isocyanate
4-bromophenyl isocyanate
4-fluorophenyl isocyanate
4-chlorophenyl isocyanate
4-methoxyphenyl isocyanate
ethyl 4-isocyanatobenzoate
4-(trifluoromethyl)phenyl isocyanate
p-tolyl isocyanate
n-(chlorocarbonyl) isocyanate
benzoyl isocyanate
tert-butyl isocyanate
(s)-(-)-alpha-methylbenzyl isocyanate
isopropyl isocyanate
methyl isocyanate
ethyl isocyanatoacetate
octadecyl isocyanate
ethyl isocyanate
2-chloroethyl isocyanate
allyl isocyanate
n-propyl isocyanate
butyl isocyanate
cyclohexyl isocyanate
1-naphthyl isocyanate
(r)-(-)-1-(1-naphthyl)ethyl isocyanate
4-fluoro-3-nitrophenyl isocyanate
2-nitrophenyl isocyanate
3-nitrophenyl isocyanate
4-nitrophenyl isocyanate
2,6-diisopropylphenyl isocyanate
benzyl isocyanate
3-chloropropyl isocyanate
ethoxycarbonyl isocyanate
3,5-bis(trifluoromethyl)phenyl isocyanate
2,4,6-tribromophenyl isocyanate
2,5-difluorophenyl isocyanate
2,4,5-trichlorophenyl isocyanate
2,4,6-trichlorophenyl isocyanate
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2-methoxycarbonylphenyl. isocyanate
2-ethoxycarbonylphenyl isocyanate
2-isopropylphenyl isocyanate
2,3-dimethylphenyl isocyanate
4-methoxy-2-methylphenyl isocyanate
2,4-dimethylphenyl isocyanate
2,5-dimethylphenyl isocyanate
2-ethyl-6-methylphenyl isocyanate
3-cyanophenyl isocyanate
5-chloro-2,4-dimethoxyphenyl isocyanate
3-chloro-4-methylphenyl isocyanate
3,5-dichlorophenyl isocyanate
5-chloro-2-methoxyphenyl isocyanate
3,4,5-trimethoxyphenyl isocyanate
3,5-dimethoxyphenyl isocyanate
3-(methylthio)phenyl isocyanate
3-ethoxycarbonylphenyl isocyanate
3-acetylphenyl isocyanate
3,4-dimethylphenyl isocyanate
3,5-dimethylphenyl isocyanate
2-methoxy-5-methylphenyl isocyanate
3-ethylphenyl isocyanate
4-chloro-2-methoxyphenyl isocyanate
4-chloro-2-trifluoromethylphenyl isocyanate
4-chloro-3-trifluoromethylphenyl isocyanate
4-iodophenyl isocyanate
4-phenoxyphenyl isocyanate
4-ethoxyphenyl isocyanate
4-(methylthio)phenyl isocyanate
4-acetylphenyl isocyanate
4-isopropylphenyl isocy~~nate
4-ethylphenyl isocyanat~~
4-n-butylphenyl isocyan;~te
3-(dichloromethylsilyl)propyl isocyanate
octyl isocyanate
4-methyl-3-nitrophenyl :isocyanate
4-chloro-2-nitrophenyl :isocyanate
2-methyl-4-nitrophenyl :isocyanate
4-methyl-2-nitrophenyl :isocyanate
2-fluoro-5-nitrophenyl :isocyanate
2-methyl-5-nitrophenyl isocyanate
3-bromopropyl isocyanatE~
2,4,6-trimethylphenyl i;;ocyanate
2-isopropyl-6-methylphenyl isocyanate
2,6-diethylphenyl isocy<~nate
5-chloro-2-methylphenyl isocyanate
4-chloro-2-methylphenyl isocyanate
4-(trifluoromethoxy)phenyl isocyanate
4-trifluoromethylthiophenylisocyanate
2,4-dibromophenyl isocy~~nate
2,6-dibromo-4-ethylphen~~l isocyanate
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2,3,4,5-tetrachlorophenyl isocyanate
2-chloro-5-trifluoromethylphenyl isocyanate
2-chloro-6-methylphenyl isocyanate
2-n-carbobutoxyphenyl isocyanate
2,4,5-trimethylphenyl isocyanate
2-methyl-6-(t-butyl)phenyl isocyanate
2-ethyl-6-isopropylphenyl isocyanate
3-chloro-2-methoxyphenyl isocyanate
3-chloro-2-methylphenyl isocyanate
3-chloro-4-fluorophenyl isocyanate
4-cyanophenyl isocyanate
4-bromo-2-methylphenyl isocyanate
4-bromo-2,6-dimethylphenyl isocyanate
2,6-dibromo-4-fluorophenyl isocyanate
4-n-butoxyphenyl isocyanate
4-butoxycarbonylphenyl isocyanate
phenethyl isocyanate
2-methyl-3-nitrophenyl isocyanate
hexyl isocyanate
hexadecyl isocyanate
methylene bis(o-chlorophenyl isocyanate)
4-chloro-3-nitrophenyl isocyanate
2-chloro-4-nitrophenyl isocyanate
4,5-dimethyl-2-nitrophenyl isocyanate
2-chloro-5-nitrophenyl isocyanate
2-methoxy-4-nitrophenyl isocyanate
3-fluoro-4-methylphenyl isocyanate
5-fluoro-2-methylphenyl isocyanate
3,5-dicarbomethoxyphenyl isocyanate
2,4-dichlorobenzyl isocyanate
2-(methylthio)phenyl isocyanate
n-(methoxycarbonyl)isocyanate
n-(phenoxycarbonyl)isocyanate
2-biphenylyl isocyanate
3-iodophenyl isocyanate
4-phenylphenyl isocyanate
tetrahydro-2-pyranyl isocyanate
4-(tert-butyl)phenylisocyanate
1-(4-bromophenyl)ethyl isocyanate
isocyanatoacetic acid n-butyl ester
dodecyl iqocyanate
6,7-methylenedioxy-4-isocyanate-methylcoumarin
(r)-(+}-alpha-methylbenzyl isocyanate
(+/-)-I-(1-naphthyl)ethyl isocyanate
(s)-(+)-1-(1-naphthyl)ethyl isocyanate
3,4-difluoropher,yl isocyanate
2-methoxy-5-nitrophenyl isocyanate
undecyl isocyanate
ethyl 2-isocyanato-4-methyl valerate
ethyl 6-isocyanatohexanoate
ethyl 2-isocyanato-4-methylthiobutyrate
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ethyl 2-isocyanatoprop_~onate
ethyl 3-isocyanatopropionate
ethyl 2-isocyanato-3-methylbutyrate
tert-butyl 3-isothiocyanatopropionate
ethyl 2-isocyanato-3-phenylpropionate
1,3-bis(isocyanatom ethyl)cyclohexane
2-(trifluoromethoxy)phenyl isocyanate
4-(chloromethyl) phenyl. isocyanate
1-adamantyl isocyanate
1,3-bis(2-isocyanato-2-~propyl)benzene
n-amyl isocyanate
n-heptyl isocyanate
2-chloroethyl isocyanat.e, [ethyl-1,2-14c]
1,1,3,3-tetramethylbutyl isocyanate
3,5-dinitrophenyl isocyanate,
and the like.
Aldehydes:
Ethyl 2-formyl-1-cyclopropanecarboxylate
Cyclohexanecarboxaldehyde
1,2,3,6-Tetrahydrobenzaldehyde
Diphenylacetaldehyde
2-Phenylpropionaldehyde
2,3-Dimethylvaleraldehyde
Isobutyraldehyde
2,6-Dimethyl-5-hepten-1-al
2-Methylbutyraldehyde
2-Ethylbutyraldehyde
2-Methylpentanal
2-Ethylhexanal
2-Methylundecanal
Phenylacetaldehyde
Isovaleraldehyde
7-Methoxy-3,7-dimethyloctanal
Undecanal
Dodecanal
Tridecanal
Tetradecyl aldehyde
Propionaldehyde
3-Phenylpropionaldehyde
3-(Methylthio)propionaldehyde
Butyraldehyde
Cis-4-decen-1-al
N-valeraldehyde
Hexanal
Heptaldehyde
Octanal
Nonanal
Decanal
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Undecylenic aldehyde
Cis-11-hexadecenal
Cis-13-octadecenal
Cis-9-hexadecenal
2,5-Dimethoxy-3-tetrahydrofurancarboxaldehyde
3,5,5-Trimethylhexanal
Succinic semialdehyde
(+/-)-3-Phenylbutyraldehyde
2,6,6-Trimethyl-1-cyclohexene-1-acetaldehyde
Cyclopropanecarboxaldehyde
3-Cyclohexylpropionaldehyde
Hydroxycitronellal
Cis-4-heptenal
Cis-6-nonen-1-al
Tetrahydrocitral
Cis-7-decen-1-al
Cis-8-undecen-1-al
3,5,6-Trimethyl-3-cyclohexene-1-carboxaldehyde
Lyral(r)
Bis(2-chlorophenyl)acetaldehyde
2-Thioglyceraldehyde
3-(4-Isopropylphenyl)isobutyraldehyde
2-Ethyl-3-methylbutanal
2-Ethylcaprylaldehyde
3-Methylvaleraldehyde
3-Phenyl-3-(p-tosyl)propionaldehyde
3-Hexenal
3-(Methylthio)butanal
Veltonal
Citronellal
2-(Trifluoromethyl)propionaldehyde
3,3-Dimethylbutyraldehyde
Campholene aldehyde
2-Formylpropionic acid methyl ester
5-Hydroxypentanal
p-Methylphenylacetaldehyde
Omega-ketoheptanoic acid
4-Chlorophenylcyanoacetaldehyde
Hexadecanal
Methyl 7-oxoheptanoate
Diethyl formyl succinate
4-Pregnene-20-beta-carboxaldehyde-3-one
Cis-7-tetradecenal
Cyclopentylmethanal
3,4-Dimethyl-3-cyclohexenylmethanal
2,4,6-Trimethyl-3-cyclohexen-1-carboxaldehyde
Adipic semialdehyde methyl ester
Cis-14-methyl-8-hexadecenal
Cis-3-hexen-1-al
Trans-4-decen-1-al
2,2-Dichlorooctadecanal
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2,2-Dichlorotetradecanal
2,2-Dichlorooctanal
2,2-Dichlorohexanal
(r)-(+)-Citronella)
8-Methyl-7-nonenal
2-(p-Tolyl)propionaldehyde
Aldehyde C-11 MOA (2-mE~thyldecanal)
Alpha-methylhydrocinnarnaldehyde
(s)-(-)-Citronella)
4-Hydroxybutanal
4-Oxobutyric acid meth~rl ester
3,3,4,4,5,5,5-Heptafluoropentanal
3-Methylbutanal-1-13c
6-Methyl-3-cyclohexene--1-carboxaldehyde
4-(4-Methyl-2-pentenyl)-3-cyclohexene-1-
carboxaldehyde
3-Pentyn-1-al
3-Pyridylacetaldehyde rf-oxide
2,3-Dihydro-5-methoxy-~~-phenyl-2-
indolecarboxaldehyde
2,4-biphenyl-3-oxobutyraldehyde
3,3,3-Triphenylpropionaldehyde
2-Bromo-n-(3-formyl-1-methylpropyl)benzamide
3-(Phenylthio)butyraldehyde
Diethyl 2-(diethoxymethyl)-3-formylsuccinate
2-Chloro-3-(4-nitrophenyl)-propionaldehyde
2-Acetoxypropionaldehycle
2-Methyl-4-phenylpentan.al
(lr,2s,3r,4s)-(+)-2-Ben.zyloxy-3-formyl-oxybornane
5-(4'-Chlorophenoxy)-1-pentanal
Boc-ala-CHO
Boc-leu-CHO
Boc-phe-CHO
Boc-tyr(OBzl)-CHO
Boc-tyr(OMe)-CHO
Boc-val-CHO
4-Pentenal
1-Formyl-6-(dimethylamino)fulvene
1,4-Dioxaspiro(4.5)decane-7-acetaldehyde
Alpha-citronella)
Diethyl 2-Acetamido-2-(2-formylethyl)malonate
3,4,4,5,5,5-Hexafluoro-3-(trifluoromethyl)pentanal
3,4,4,4-Tetrafluoro-3-(heptafluoropropoxy)butanal
3,4,4,4-Tetrafluoro-3-(trifluoromethoxy)butanal
3,4,4,4-Tetrafluoro-3-(trifluoromethyl)butanal
3,3,4,4,5,5,6,6,7,7,8,8,8-Tridecafluorooctanal
3,3,3-Trifluoropropanal
Beta,beta-dimethylhydrocinnamaldehyde
5-Norbornene-2-carboxaldehyde
Chrysanthal
9-Decenal
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Decyl aldehyde, [1-14c]
4,4,4-Trifluorobutyraldehyde
3-Methyl-3-butenal
3-(5-Methyl-2-furyl)butanal
3-Phenyl-4-pentenal
Tert-butyl (s)-4-formyl-2,2-dimethyl-3-
oxazolidinecarboxylate
Trans-2-dodecenal
9,10-Dihydro-9,10-ethanoanthracene-11-carboxaldehyde
Methyl hexyl acetaldehyde
2,3-Dihydro-2-oxo-1H-imidazol-4-carboxaldehyde
N-Acetylmuramic acid,
and the like.
Other suitable aldehydes useful in preparation of
the present libraries are further illustrated by the
following formulas, wherein L is -CHO:
L\ ~ i
v
L
O / 1 L i L
/ 1 ~ o / ~ ' ~ o
/ \ i
L L L
i
la _ N.~ o
~ CH3
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L L L
O
a
/ / C~ /
O
OH
OH
L L
~C/ O \
i
/ ~ C H
/ o
H3C~O~ O~ CH3 OH
L L L
OH °' a
i I \
.-- /
OH
s
H,c ~
L L
L
OH
O O
1 _ I .
N
H3C~ ~ CHI
L L
HD
O OH
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_7p_
L
HN, ~ I ~ I
HN CH3
O
L L
( ~ I I \
~c~ ~ ~ a~
0 0
Q"I C H 3
N O O~
L L
\
' /
ci I - Br
F
L L
\
f i ~ c~ \
0
H3C O / CI
o CI CH3
L
L
\ ~ ~c
o = . / ~ cH3
'o
CH3 0
off B
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L O L
i i / O ~ /
Br
L L
CH3
F /
I
O
L L L
Br
F nl
L L L
O~N ~ ~ W /
O
O O
O\
C H3
L
F
o' _cH, F
~0
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L L L
I ~ ~ ~ F
/ / /
I~C~ N~ CI~
F
L L L
i
i
L
CI
OH
L
L
L
n
O CHs
CH3
,3 S L
HaC L
C H3
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L
f"~C O CJ"~ I w
I
O CHs
L L L
li '
\ i
H3C o
CH3
L o t L
~C i ' I I _ I
HsC ~ / ~ I ~ o
~' ~ o
H3C '
L
O L L
I . '
i ~ I I
'3C~° H3C ~ /
V
H3 C
CH3
L
' L
L
I ~ o II
0
w ~C~/~/~ O
HOC O
I
L L
H3C / L
H3~~ J ~ ~
H3 C O I
CH3
L L L
a
I . . I C~
0
0
roc
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L L
L
1
w ~ ~u ~ ~ /J
V
i
Br L
H3C~
O
/O
N3C
Ketones:
1-BROMO-2-BUTANONE
2-FLUOROPHENYLACETONE
4-FLUOROPHENYLACETONE
3-TRIFLUOROMETHYLPHENYLACETONE
1,1,1-TRIFLUORO-2,4-PENTANEDIONE
FLUOROACETONE
4'-CHLOROACETOACETANILIDE
3-CHLORO-2-BUTANONE
CHLOROACETONE
1,3-DICHLOROACETONE
METHYL 4-CHLOROACETOACETATE
ETHYL 4-CHLOROACETOACETATE
1-CHLORO-3-PENTANONE
5-CHLORO-2-PENTANONE
CYCLOTRIDECANONE
CYCLOPENTADECANONE
DICYCLOPROPYL KETONE
CYCLOPROPYL METHYL KETONE
(-)-THUJONE
CYCLOBUTANONE
2-CYCLOPENTEN-1-ONE
3-METHYL-2-CYCLOPENTEN-1-ONE
4-CYCLOPENTENE-1,3-DIONE
1,3-CYCLOPENTANEDIONE
2-METHYL-1,3-CYCLOPENTANEDIONE
CYCLOPENTANONE
2-CHLOROCYCLOPENTANONE
CYCLOPENTANONE-2-CARBOXYLIC ACID METHYL ESTER
ETHYL 2-OXOCYCLOPENTANECARBOXYLATE
2-ACETYLCYCLOPENTANONE
2-METHYLCYCLOPENTANONE
2,4-DIMETHYLCYCLOPENTANONE
3-METHYLCYCLOPENTANONE
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BETA-IONONE
BETA-IONONE
ALPHA-IONONE
2-CYCLOHEXEN-1-ONE
3-ETHOXY-2-CYCLOHEXEN-1-ONE
3-METHYL-2-CYCLOHEXEN-:1-ONE
4-CARBETHOXY-3-METHYL-7-CYCLOHEXEN-1-ONE
3,5-DIMETHYL-2-CYCLOHE:KEN-1-ONE
ISOPHORONE
1,3-CYCLOHEXANEDIONE
2-ACETYL-1,3-CYCLOHEXAI~EDIONE
2-METHYL-1,3-CYCLOHEXAI~TEDIONE
DIMEDONE
1,4-CYCLOHEXANEDIONE
DIMETHYL 1,4-CYCLOHEXANEDIONE-2,5-DICARBOXYLATE
DIETHYL 1,4-CYCLOHEXANEDIONE-2,5-DICARBOXYLATE
CYCLOHEXANONE
2-CHLOROCYCLOHEXANONE
2-NITROCYCLOHEXANONE
2-PHENYLCYCLOHEXANONE
2-(3-METHOXYPHENYL)CYCI~OHEXANONE
ETHYL 2-CYCLOHEXANONEC~~RBOXYLATE
ETHYL 4-METHYL-2-CYCLOFiEXANONE-1-CARBOXYLATE
2-ACETYLCYCLOHEXANONE
L-MENTHONE
2-METHYLCYCLOHEXANONE
(+)-DIHYDROCARVONE
2,6-DIMETHYLCYCLOHEXANC)NE
ETHYL-2-CYCLOHEXANONE ACETATE
3-METHYLCYCLOHEXANONE
3,3,5,5-TETRAMETHYLCYCI~OHEXANONE
4-PHENYLCYCLOHEXANONE
4-TERT-BUTYLCYCLOHEXANONE
4-METHYLCYCLOHEXANONE
4-ETHYLCYCLOHEXANONE
3,4,8,8A-TETRAHYDRO-8A-METHYL-1,6(2H,7H)-
NAPHTHALENEDIONE
2-ACETYL-1-TETRALONE
1-DECALONE
BETA-TETRALONE
1-METHYL-2-TETRALONE
6-METHOXY-2-TETRALONE
7-METHOXY-2-TETRALONE
2-DECALONE
CYCLOOCTANONE
CYCLONONANONE
ALPHA-ACETYL-PHENYLACETONITRILE
ANTI-PYRUVIC ALDEHYDE 1-OXIME
4-(4-HYDROXYPHENYL)-2-BUTANONE
1,3-ACETONEDICARBOXYLIC ACID
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LEVIJLINIC ACID
4,6-DIOXOHEPTANOIC ACID
4-KETOPIMELIC ACID
3-ACETYL-1-PROPANOL
4-ANDROSTENE-3,17-DIONE
CYCLODECANONE
CYCLOUNDECANONE
CYCLODODECANONE
CIS-BICYCLO(3.3.0)OCTANE-3,7-DIONE
CIS-1,5-DIMETHYLBICYCLO(3.3.0)OCTANE-3,7-DIONE
2-INDANONE
2-CYCLOHEPTEN-1-ONE
CYCLOHEPTANONE
4-ACETYLBUTYRIC ACID
4-HYDROXY-4-METHYL-2-PENTANONE
3-HYDROXY-2-BUTANONE
HYDROXYACETONE
4-HYDROXY-3-METHYL-2-BUTANONE
1-BENZYL-3-PYRROLIDINONE
2-ACETYLBUTYROLACTONE
TETRAHYDROTHIOPHEN-3-ONE
TROPINONE
6-HYDROXYTROPINONE
4-OXO-TEMPO
N-CARBETHOXY-4-PIPERIDONE
N-BENZOYL-4-PIPERIDONE
1-ACETYL-4-PIPERIDONE
1-METHYL-4-PIPERIDONE
1-BENZYL-4-PIPERIDONE
1-(BETA-PHENYLETHYL)-4-PIPERIDONE
TETRAHYDRO-4H-PYRAN-4-ONE
1,4-CYCLOHEXANEDIONE MONO-2,2-
DIMETHYLTRIMETHYLENE
KETAL
TETRAHYDROTHIOPYRAN-4-ONE
METHYLGLYOXAL
DIMETHYL ACETYLSUCCINATE
DIMETHYL 1,3-ACETONEDICARBOXYLATE
DIMETHYL 3-OXOADIPATE
4-(4-ACETOXYPHENYL)-2-BUTANONE
METHYL 2-CHLOROACETOACETATE
METHYLGLYOXAL DIMETHYL ACETAL
1,1-DIPHENYLACETONE
1,1-DIPHENYLACETONE
METHYL 4-ACETYL-5-OXOHEXANOATE
3-METHYLTHIO-2-BUTANONE
3-METHYLTHIO-2-BUTANONE
3-METHYL-2,4-PENTANEDIONE
3-ETHYL-2,4-PENTANEDIONE
ACETONE
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(DIMETHYLAMINO)ACETONE
PHENOXYACETONE
METHOXYACETONE
DIMETHYL (2-OXOPROPYL)F'HOSPHONATE
2-METHOXYPHENYLACETONE
3-METHOXYPHENYLACETONE
3,4-DIMETHOXYPHENYLACE'TONE
4-METHOXYPHENYLACETONE
4-METHOXYPHENYLACETONE
METHYL VINYL KETONE
4-METHOXY-3-BUTEN-2-ONE.
4-METHOXY-3-BUTEN-2-ONE.
BENZYLIDENEACETONE
BENZYLIDENEACETONE
ACETOACETANILIDE
O-ACETOACETANISIDIDE
O-ACETOACETOTOLUIDIDE
P-ACETOACETANISIDIDE
METHYL ACETOACETATE
BENZYL ACETOACETATE
BENZOYLACETONE
ACETYLACETONE
DIACETONE ACRYLAMIDE
ACETYLACETALDEHYDE DIMETHYL ACETAL
BENZYLACETONE
4-(4-METHOXYPHENYL)-2-BUTANONE
ACETONYLACETONE
5-HEXEN-2-ONE
N-TERT-BUTYLACETOACETAMIDE
TERT-BUTYL ACETOACETATE
4,4-DIMETHYL-2-PENTANONE
MESITYL OXIDE
PHORONE
6-METHYL-5-HEPTEN-2-ONE
GERANYLACETONE
GERANYLACETONE
DIISOPROPYL KETONE
3-METHYL-2-BUTANONE
METHYL ISOBUTYL KETONE
6-METHYL-2,4-HEPTANEDIOIVE
2,6-DIMETHYL-4-HEPTANONE
5-METHYL-2-HEXANONE
2-TRIDECANONE
DTETHYLAMINOACETONE
1-DIETHYLAMINO-3-BUTAN02JE
S-DIETHYLAMINO-2-PENTANONE
ETHYL 2,4-DIOXOVALERATE
ETHYL 2-CHLOROACETOACETATE
ETHYL 2-BENZYLACETOACETATE
DIETHYL ACETYLSUCCINATE
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DIETHYL 2-ACETYLGLUTARATE
ETHYL 2-METHYLACETOACETATE
ETHYL ISOBUTYRYLACETATE
ETHYL ACETOACETATE
DIETHYL 1,3-ACETONEDICARBOXYLATE
ETHYL LEVULINATE
DIETHYL 4-OXOPIMELATE
ETHYL 4-ACETYL-5-OXOHEXANOATE
ETHYL 4-ACETYLBUTYRATE
DIETHYL 3-OXOPIMELATE
3-PENTEN-2-ONE
2-METHYL-3-PENTANONE
1-PHENYL-2-BUTANONE
ETHYL VINYL KETONE
ETHYL PROPIONYLACETATE
2,4-HEXANEDIONE
3-PENTANONE
3-METHYL-2-PENTANONE
5-METHYL-3-HEPTANONE
2-METHYL-3-HEXANONE
2-PENTANONE
ETHYL BUTYRYLACETATE
3-HEXANONE
3-HEXANONE
4-HEPTANONE
BUTYL LEVULINATE
2-METHYL-3-HEPTANONE
2-HEXANONE
3-HEPTANONE
5-NONANONE
2-HEPTANONE
DIMETHYL (2-OXOHEPTYL)PHOSPHONATE
3-OCTANONE
6-UNDECANONE
2-OCTANONE
3-NONANONE
4-DECANONE
7-TRIDECANONE
2-NONANONE
3-DECANONE
8-PENTADECANONE
2-DECANONE
3-UNDECANONE
9-HEPTADECANONE
2-UNDECANONE
10-NONADECANONE
2-METHOXYCYCLOHEXANONE
2-METHOXYETHYL ACETOACETATE
3-CHLOROACETYLACETONE
1,1-DICHLOROACETONE
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6,7-DIMETHOXY-2-TETRALC)NE
4,4-DIMETHYL-2-CYCLOHE~;EN-1-ONE
2-TERT-BUTYLCYCLOHEXANCINE
N-(ACETOACETYL)GLYCINE
3,5-DIACETYLTETRAHYDROF'YRAN-2,4,6-TRIONE
ALLYL ACETOACETATE
4-(TRIMETHYLSILYLOXY)-3-PENTEN-2-ONE
ETHYL DIACETOACETATE
(R)-(-)-4,4A,5,6,7,8-HE~XAHYDRO-4A-METHYL-
2(3H)NAPHTHALENONE
4,4-DIMETHOXY-2,5-CYCLOHEXADIEN-1-ONE
3-OXOADIPIC ACID
TETRONIC ACID
METHYL 4-METHOXYACETOACETATE
DI-TERT-BUTYL 1,3-ACETONEDICARBOXYLATE
1,4-CYCLOHEXANEDIONE MONOETHYLENEKETAL
DIMETHYL (3-PHENOXYACETONYL)PHOSPHONATE
4-ACETOXY-2-BUTANONE
3-NONEN-2-ONE
1-HYDROXY-2-BUTANONE
5-METHYL-1,3-CYCLOHEXANEDIONE
2-METHYLTETRAHYDROFURAN-3-ONE
1-PROPYL-4-PIPERIDONE
5ALPHA-ANDROST-26-EN-3-ONE
2-CHLORO-5,5-DIMETHYL-1,3-CYCLOHEXANEDIONE
4-ACETYL-1-METHYL-1-CYCLOHEXENE
4-ACETYL-1-METHYL-1-CYCLOHEXENE
4-HEXEN-3-ONE
4-(TERT-BUTYLDIMETHYLSILYLOXY)-3-PENTEN-2-ONE
5-KETOHEXANENITRILE
ETHYL 2-(TRIMETHYLSILYL:METHYL)ACETOACETATE
ETHYL 4-HYDROXY-6-METHYL-2-OXO-3-CYCLOHEXENE-
1-CARBOXYLATE
METHYL 4-ALLYL-3,5-DIOXO-1-CYCLOHEXANECARBOXYLATE
METHYL 2-OXO-1-CYCLOHEP'rANECARBOXYLATE
METHYL ETHYL KETONE
METHYL 3-OXOPENTANOATE
5-METHYL-3-HEXEN-2-ONE
4-(4-HYDROXY-3-METHOXYPHENYL)-3-BUTEN-2-ONE
(S)-(+)-2,3,7,7A-TETRAH'YDRO-7A-METHYL-1H-INDENE-
1,5(6H)-DIONE
2-HYDROXYMETHYL-6-METHO:?~Y-1,4-BENZOQUINONE
8-CYCLOHEXADECEN-1-ONE
8-MERCAPTOMENTHONE
2,2,6,6-TETRAMETHYL-4-P:LPERIDONE HYDROCHLORIDE
METHYL 4-OXO-3-PIPERTDIIJECARBOXYLATE
HYDROCHLORIDE
ETHYL 1-BENZYL-3-OXOPIP1~RIDINE-4-CARBOXYLATE
HYDROCHLORIDE
1-ETHYL-3-PIPERIDONE HYI7ROCHLORIDE
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METHYL 1-BENZYL-4-OXO-3-PIPERIDINE-CARBOXYLATE
HYDROCHLORIDE
1-BENZYL-3-CARBETHOXY-4-PIPERIDONE HYDROCHLORIDE
DELTA-AMINOLEWLINIC ACID, METHYL ESTER
HYDROCHLORIDE
5-AMINOLEVLJLINIC ACID HYDROCHLORIDE
1,4-DIAMINO-2-BUTANONE DIHYDROCHLORIDE
DIACETONAMINE HYDROGEN OXALATE
ACETOACETIC ACID LITHIUM SALT
S-TERT-BUTYL ACETOTHIOACETATE
ETHYL 4-OXOCYCLOHEXANECARBOXYLATE
1-ETHYL-4-PIPERIDONE
N-HYDROXYSUCCINIMIDYL ACETOACETATE
3-BROMO-2-BUTANONE
1,3-DIBROMOACETONE
(4-CHLOROPHENYLTHIO)PROPAN-2-ONE
3-AMINO-2-CYCLOHEXEN-1-ONE
2-{1-CYCLOHEXENYL}CYCLOHEXANONE
2-(BETA-CYANOETHYL)CYCLOHEXANONE
4-TERT-AMYLCYCLOHEXANONE
3-ACETYLACRYLIC ACID
TROPONE
METHYLSULFONYLACETONE
BIS(4-METHOXYBENZYLIDENE)ACETONE
BIS(4-METHOXYBENZYLIDENE)ACETONE
3,4-DIMETHOXYBENZYLIDENEACETONE
5-METHYL-5-HEXEN-2-ONE
2,6-DIMETHYL-3,5-HEPTANEDIONE
2-DODECANONE
3,5-HEPTANEDIONE
2,4-OCTANEDIONE
2,4-NONANEDIONE
5-DODECANONE
4-DODECANONE
3-DODECANONE
(METHYLTHIO)ACETONE
(+/-)-2-ETHOXYCYCLOHEXANONE
3-HEPTEN-2-ONE
3-OCTEN-2-ONE
2,6-DIMETHOXY-P-BENZOQUINONE
4-(P-HYDROXYPHENYL)-3-BUTEN-2-ONE
3,4-METHYLENEDIOXYBENZYLIDENE ACETONE
3,4-METHYLENEDIOXYBENZYLACETONE
PHENYLACETONE
2',5'-DICHLOROACETOACETANILIDE
4-CHLOROPHENYLSULFONYLACETONE
4-CHLOROBENZYLIDENEACETONE
2,4,4-TRIMETHYLCYCLOPENTANONE
CYCLOHEXYLACETONE
3,3,5-TRIMETHYLCYCLOHEXANONE
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4-PYRAZOLINO-2-BUTANONE
ACETOACETAMIDE
3-PHENYLAZOACETYLACETONE
3 -N-BUTYL-2 , 4-PENTANED7:ONE
3-ACETYLOCTANONE-2
SALOR 517,446-7
(BENZYLTHIO)ACETONE
6-PHENYLHEXA-3,5-DIEN-2-ONE
3-TRIDECANONE
5-PENTADECANONE
7-HEPTADECANONE
4-HEPTADECANONE
3-OCTADECANONE
N,N-DIETHYLACETOACETAMIDE
ETHYL 2-(PHENYLAZO)ACETOACETATE
DIETHYL ACETOMALONATE
ETHYL-2-N BUTYLACETOACETATE
(ETHYLTHIO)ACETONE
1-PHENYL-2-PENTANONE
1-PHENYL-2-HEXANONE
N-AMYL ISOPROPYL KETONE
7-PENTADECANONE
4-TRIDECANONE
5-TETRADECANONE
6-PENTADECANONE
7-HEXADECANONE
1-OCTEN-3-ONE
NOOTKATONE
CYCLOBUTYL METHYL KETONE
1-ISOPROPYL-4-PIPERIDONE
N,N-DIMETHYLACETOACETAM=CDE
FURFURALACETONE
ACETOACET-M-XYLIDIDE
ETHYL 2 -ETHYLACETOACETA'l'E
DICYCLOHEXYL KETONE
CYCLOHEXYL METHYL KETONE
1,2:5,6-DI-O-ISOPROPYLII)ENE-ALPHA-D-RIBO-3-
HEXOFURANOSULOSE
4-HYDROXYPYRIDINE
BENZYL ISOPROPYL KETONE
1,1,3-TRICHLOROACETONE
METHYL BUTYRYLACETATE
CHLORODIFLUOROACETYLACETONE
1-(2-FURYL)-1,3-BUTANEDIONE
4,6-NONANEDIONE
2-HEXADECANONE
N,N-DIMETHYL-2-CHLOROACETOACETAMIDE
3-ETHOXY-2-CYCLOPENTEN-1-ONE
2,6,6-TRIMETHYL-2-CYCLOHEXENE-1,4-DIONE
ETHYL 4-PIPERIDONE-3-CARBOXYLATE HYDROCHLORIDE
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CYCLOPENTYLACETONE
4-ISOPROPYLCYCLOHEXANONE
(+/-)-2-ALLYLCYCLOHEXANONE
6-METHYL-3,5-HEPTADIEN-2-ONE
FLUORAL-P
FLUORAL-P
ISOPROPYL ACETOACETATE
N-METHYLACETOACETAMIDE
2-ETHYL-1,3-CYCLOPENTANEDIONE
3,5-DIMETHYL-4-HEPTANONE
EXO-2-CHLORO-5-OXO-BICYCLO[2.2.1]HEPTANE-SYN-
7-CARBOXYLIC ACID
ETHYL 2-OXO-CYCLOPENTYLACETATE
ETHYL-6-(2-OXOCYCLOPENTYL)-HEXANOATE
DIETHYL (2-OXOPROPYL)PHOSPHONATE
MAYBRIDGE BTBG 0108
4-ACETYLPIPERIDINE HYDROCHLORIDE
4-CYANO-4-PHENYLCYCLOHEXANONE
1,1,1-TRIFLUORO-5-METHYL-2,4-HEXANEDIONE
O-(CHLOROPHENYL)ACETONE
3-AMINO-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE
PHENYL SULFONYL ACETONE
1,3-DIPHENYL-2,4-PENTANEDIONE
3-ACETYLHEXANONE-2
1-PHENYL-2,4-PENTANEDIONE
3-HEXADECANONE
S-UNDECANONE
2-METHYL-3-DECANONE
4-UNDECANONE
6-TRIDECANONE
5-TRIDECANONE
1,1,1-TRIFLUORO-2,4-HEXANEDIONE
2-CYCLOPENTYLCYCLOPENTANONE
2-SEC-BUTYLCYCLOHEXANONE
2-BENZYLCYCLOHEXANONE
5-ACETYLVALERIC ACID
4-NITROPHENYL ACETONE
6-HEXADECANONE
5-HEXADECANONE
6-TETRADECANONE
1-HEXEN-3-ONE
2-CHLORO-6-FLUOROBENZYLIDENEACETONE
MAYBRIDGE CD 09843
3-CHLORO-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE
2-ACETOXY-3-BUTANONE
5-PHENYLCYCLOHEXANE-1,3-DIONE
TERT.-BUTYL-4-CHLOROACETOACETATE
ISOPROPYL 4-CHLOROACETOACETATE
3-METHYLENE-2,6-HEPTANEDIONE
(4-METHYLPHENYLTHIO)ACETONE
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1-(THIEN-2-YL)BUT-1-EN-3-ONE
2-PENTADECANONE
ACETOACETOXYETHYL METHACRYLATE
4-PROPYLCYCLOHEXANONE
4-HYDROXY-2-BUTANONE
METHYL TRAMS-4-OXO-2-PI~NTENOATE
METHYL TRAMS-4-OXO-2-PI~NTENOATE
ISOBUTYL ACETOACETATE
2-N-HEXYLCYCLOPENTANONE
3-FLUOROPHENYLACETONE
BENZYL LEVULINATE
(R)-(+)-3-METHYLCYCLOHEXANONE
(R)-(+)-3-METHYLCYCLOPENTANONE
TRAMS-1-DECALONE
(S)-{+)-10-METHYL-1(9)--OCTAL-2-ONE
(1S)-{-)-VERBENONE
DEHYDROACETIC ACID
PINONIC ACID
PINONIC ACID
1,3-DIAMINOACETONE DIHS.'DROCHLORIDE MONOHYDRATE
(5S)-5,6-ISOPROPYLIDENE:DIOXY-6-METHYL-HEPTAN-2-
ONE
N-OCTYL 4-CHLOROACETOAC.'ETATE
MAYBRIDGE KM 02248
BICYCLO[3.2.1]OCTAN-2-ONE
2-ADAMANTANONE
BICYCLO(3.3.1)NONAN-9-ONE
BICYCLO(3.3.1)NONANE-3,7-DIONE
(1R)-(+)-NOPINONE
NORCAMPHOR
NORCAMPHOR
NERYLACETONE
4,4-DIMETHYL-2-CYCLOPEN'TEN-1-ONE
4,4-DIPHENYL-2-CYCLOHEXEN-1-ONE
THIOTETRONIC ACID
4,4-DIMETHYL-1,3-CYCLOHEXANEDIONE
2-CHLORO-1,4-BENZOQUINONE
7-OXOOCTANOIC ACID
METHYL 3-OXO-6-OCTENOATE
5,5-DIMETHYL-2-PHENACYL-1,3-CYCLOHEXANEDIONE
5-OXOAZELAIC ACID
3-(2-HYDROXYETHYLAMINO)-5,5-DIMETHYL-2-
CYCLOHEXEN-
1-ONE
ABSCISIC ACID
ABSCISIC ACID
N-CARBETHOXY-4-TROPINONE
2-METHYLTETRAHYDROTHIOPHEN-3-ONE
CYCLOHEXIMIDE
PSEUDOPELLETIERINE HYDROCHLORIDE
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DEHYDROCARNITINE HYDROCHLORIDE
3-(PHENYLAMINO)-CYCLOHEX-2-ENE-1-ONE
3-HEPTADECANONE
EXO-2-BROMO-5-OXO-BICYCLO[2.2.1]HEPTANE-SYN-7-
CARBOXYLIC ACID
ANTI-3-OXOTRICYCLO(2.2.1.02,6)HEPTANE-7-
CARBOXYLIC
ACID
(+/-)-ISOPHORONE OXIDE
PHTHALIMIDOACETONE
1-PHENYL-1,4-PENTANEDIONE
(+/-)-2-PHENYLCYCLOHEPTANONE
ACETYLMALONONITRILE
4-HYDROXY-3-METHOXYPHENYLACETONE
5,7-DIMETHYL-3,5,9-DECATRIEN-2-ONE
ETHYL 6-METHYL-2-OXO-3-CYCLOHEXENE-1-CARBOXYLATE
3-CHLOROTETRONIC ACID
2,4-DIHYDRO-5-METHYL-2-PHENYL-4-PROPIONYL-3H-
PYRAZOL-3-ONE
D-(-)-TAGATOSE
3-(DIMETHYLAMINO)-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE
(3AS,7AS)-(+)-HEXAHYDRO-3A-HYDROXY-7A-METHYL-
1,5-INDANDIONE
(+/-)-EXO-6-HYDROXYTROPINONE
(1R-(1ALPHA,2BETA,3ALPHA))-(+)-3-METHYL-
2-(NITROMETHYL)-5-OXOCYCLOPENTANEACETIC
MENTHONE
3-QUINUCLIDINONE HYDROCHLORIDE
1,5-DIAMINO-3-OXAPENTANE
1-DIMETHYLAMINO-BUT-1-EN-3-ONE
(1R,3S)-2,2-DIMETHYL-3-(2-OXOPROPYL)-
CYCLOPROPANEACETONITRILE
(1S,3S)-3-ACETYL-2,2-DIMETHYLCYCLOBUTANE
ACETONITRILE
5,5-DIMETHYLHEXANE-2,4-DIONE
3-ISOBUTOXY-2-CYCLOHEXEN-1-ONE
3-METHYL-5-METHOXYCARBONYL-1-BENZYL-4-PIPERIDONE
HYDROCHLORIDE
3-METHYL-5-METHOXYCARBONYL-4-PIPERTDONE
HYDROCHLORIDE
DL-3-(1-ACETOXY-1-METHYLETHYL)-6-
OXOHEPTANENITRILE
DL-3-(1-METHYL-1-ETHENYL)-6-OXOHEPTANENITRILE
ETHYL 2-OXO-1-CYCLOOCTANECARBOXYLATE
METHYL (1R,3S)-2,2-DIMETHYL-3-(2-OXOPROPYL}-
CYCLOPROPANEACETATE
N-BENZYLPSEUDOPELLETIERINE
D-(-)-FRUCTOSE
(S)-(+)-ERYTHRULOSE HYDRATE
2,2,6,6-TETRAMETHYL-4-PIPERIDONE MONOHYDRATE
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1-BENZYL-3-PIPERIDONE HYDROCHLORIDE HYDRATE
5-ISOPROPYL-1,3-CYCLOHE;XANEDIONE HYDRATE
D-(+)-SORBOSE
L- ( - ) -SORBOSE
METHYL JASMONATE
N-BENZYLTROPINONE
N-TERT-BUTOXYCARBONYL-4-PIPERIDONE
(+/-)-BICYCLO(3.3.1)NON.'ANE-2,6-DIONE
ANTI-5-CARBOXYTRICYCLO[2.2.1.0(2,6)]HEPTAN-3-ONE
EXO-2-CHLORO-SYN-7-HYDR.OXYMETHYL-5-OXO-
BICYCLO[2.2.1]HEPTANE
L-RIBULOSE HYDRATE
4-ACETYL-2,4-DIHYDRO-5-METHYL-2-PHENYL-3H-
PYRAZOL-3-ONE MONOHYDRATE
2-ACETYL-5-NORBORNENE
7-SYN-METHOXYMETHYL-5-NORBORNEN-2-ONE
3-CHLORO-2-NORBORNANONE
TRICYCLO[5.2.1.02,6]DECAN-8-ONE
METHYL 4-METHOXY-2-OXO-3-CYCLOPENTENE-1-
CARBOXYLATE
5,5-DIMETHYL-3-(METHYLAMINO)-2-CYCLOHEXEN-1-ONE
2-ACETYL-1,3-CYCLOPENTAI~EDIONE
2,4-DIMETHOXYPHENYLACETONE
2, 6-DIPHENYLCYCLOHEXANO:I~E
5-HYDROXY-2-ADAMANTANON:E
3-METHOXY-2-CYCLOPENTEN-1-ONE
2,2'-METHYLENEBIS(1,3-C'YCLOHEXANEDIONE)
(+/-)-7-OXABICYCLO(4.1.0)HEPTAN-2-ONE
ETHYL 2-OXOCYCLOTRIDECA7VECARBOXYLATE
2-(METHYLTHIO)CYCLOHEXA1VONE
(S)-(+)-3,4,8,8A-TETRAH'~DRO-8A-METHYL-1,6(2H,7H)-
NAPHTHALENEDIONE,
and the like.
Where aldehydes and ketones are utilized as the
electrophilic reagent, the rE~sulting imine compounds can
be reduced using sodium borohydride or other borohydride
reducing agents to provide the corresponding secondary
amine (Formula I; Ry~~ _ hydrogen and E is a substituent
derived from an electrophilic: agent). Still further
diversity can be introduced into the present library by
reacting the secondary amine library compounds with an
electrophilic reagent as described above to provide
compounds of Formula I wherein Ry~~ and E are each
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independently a substituent derived from an electrophilic
reagent.
Solid phase synthesis methods have been modified in
accordance with the present invention to provide a
general route to preparing the novel macrocycles of the
present invention (See Scheme III). In one embodiment
(bold-face numerals corresond to compound numerals in
Scheme III), a foundation was laid by anchoring to solid
support an orthogonally protected trifunctional compound
such as N-Fmoc-S-trityl-L-cysteine to give 2 (step a).
The cycle was then built up (steps b,c,d) by coupling
desired amino acid and ATA building blocks to provide
intermediates such as 5. To facilitate cyclization, a
bromoacid was installed at the terminal position (step f)
to give intermediate 6. Bromoamide 6 was then treated
trifluoroacetic acid (TFA) to liberate it from solid
support and to unmask its side chain functional groups.
Intramolecular attack of the revealed cysteine sulfhydryl
function on the terminal alpha-bromoamide then formed
macrocycle 7, a 19-membered bicycle that possesses a
number of significant backbone modifications including
two thioether linkages and the rigid 2-
mercaptonicotinamide unit whose pyridine type nitrogen
may accept hydrogen bonds.
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_87-
Trt o
Trt o N
FmocHtQ - a". N'~ bi NH H
FmocHN H
'O -
2 FmocH ~'~ ~ ~ Otbu
3
c
HFmoc
4
bu
.I
O
NH H f
----
O -
' ~ ~ Otbu
H
Scheme III
Method for assembling a specific pseudopeptide
macrocycle (shaded spheres represent a solid support):
a) Fmoc-Rink amide MBHA resin, 30% piperidine in DMF,
then 5 eq Fmoc-S-trityl-L-cysteine, DIC/HOBT in NMP; b)
as for a), but with 5 eq Fmoc-L-tyrosine-O-t-butyl
ether; c) as for a), but with 5eq Fmoc-L-Phe[y~CH2S)-2-
mercaptonicotinate; d) as for a), but with 5eq Fmoc-D-
alanine; e) 30% piperidine in DMF, then 5eq bromoacetic
acid and DIC in NMP; f) 38:1:2 TFA:H20:Et3SiH for 2h,
lyophilize, wash (ether, 2:x) then 2eq of
diisopropylethylamine in 1:1 CH3CN:H20, 24h.
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Macrocyclization kinetics of 7 is accelarated by the
addition of a base. LCMS is utilized for monitoring ring
closure. The two major natural isotopes of bromine in
linear material were plainly visible at 80 and 82 m.u.
(mass units) over the calculated mass of the cycle. Ring
closure was accompanied by a shift in HPLC retention time
and a change in the mass spectrum to a non-doubled peak
consistent with the mass calculated for cyclic product
(for 7, calculated MW=501.41, found MH+=502.2). Under
the conditions described, LCMS revealed little or no
dimer formation during closure of 7 or any of the other
macrocycles made regardless of ring size or structural
rigidity. HPLC traces of macrocycles bearing racemic
ATAs, such as compound 32 as shown in Method B below,
often displayed a pair of closely eluting peaks having
similar abundance and identical mass spectra. Thus both
ring diastereomers formed and were separable by HPLC.
By utilizing the large number of ATA building blocks
and the large number of amino acids and amino/acid
bearing compounds that are available commercially, large
numbers of unique macrocyclic pseudopeptides can be
synthesized. Approximately seven hundred macrocycles
have been made using automated multiple parallel
synthesis and representatives from this set are shown
below (numerals shown within ring structures represent
the number of macrocycle ring atoms and Bn is benzyl).
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__gg_
H ;O
~('~N
'13 S
~O
O
NHy
The synthesis method of the present invention allows
the construction of a library of about 12- to about 24-
membered macrocycles in integral steps. This capability
is important for varying molecular size, volume and
shape. Each library compound also contains at least two
thioether linkages to enhance lipophilicity and protease
resistance relative to simple cyclic peptides. The
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position of one thioether is fixed at the cysteine or
penicillamine used for ligation, but the others) can be
installed at any internal site depending on the coupling
order of the ATA and amino acid compounds. As judged by
LCMS, macrocyclization occurred readily with a variety of
L- and D-amino acids without regard to sequence within
the ring. Macrocycles containing single or multiple aryl
backbone segments also formed readily, exemplified by
compounds 14, 16, 17 and 19-24 This is notable since aryl
backbones will impart structural rigidity to macrocycles.
Even more important, ortho-(20), meta-(21) and para-
substituted (22) aryl ring backbones all permitted
efficient ring closure. Model building suggests that
macrocycle conformational bias can be influenced by the
ortho- vs. meta- vs. para-substitution pattern of the
ATAs used during construction. This is a highly desirable
feature for de novo lead generation where the active
shapes) and conformations) are unknown.
The process of the present invention utilized in
preparation of a library of the macrocycle of Formula I
above may be carried out in any vessel capable of holding
the liquid reaction medium. In one embodiment, the
process of the invention can be carried out in containers
adaptable to parallel array synthesis. In particular,
the macrocycle library of this invention can be formed in
a well plate apparatus 1 or 3 as illustrated in Figures 1
and 2, respectively, and as described in greater detail
below. Such apparatus provide multiple reaction zones
most typically in a two-dimensional array of defined
reservoirs, wherein one member of the macrocycle library
of this invention is prepared in each reservoir. Thus
the diverse macrocycle library of the present invention
comprises a plurality of reservoir arrays (e. g. well
plates), each reservoir or well containing a library
compound of the macrocycle library. Accordingly the
library compounds are typically identified by reference
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to their well plate number a;nd their X column and Y row
well plate coordinates.
Following simultaneous preparation of the library
member compounds in the reservoir array, the compounds
can be transferred in whole or in part to other reservoir
arrays (e.g. well plates), to prepare multiple copies of
the library apparatus or to subject the library to
additional reaction conditions. Copies of the library
apparatus (daughter well plates, each comprising a 2-
dimensional array of defined reservoirs with each
reservoir containing a predetermined member of the
library) are useful as replaceable elements in automated
assay machines. The apparatus of this invention allows
convenient access to a wide variety of structurally
related macrocyclic compounds. One preferred reservoir
array for use in making and using this invention is a
multi-well titer plate, typically a 96-well microtiter
plate.
Figure 1 illustrates the top surface of a well plate
apparatus (1) of the present invention. The well plate
(1) is a plastic plate with 96-wells (depressions)
capable of holding liquids for parallel array synthesis.
Individual reaction products are prepared in each well
and are labeled by the well ~?late coordinates. For
example, the library compound at location (2), is
identified by the alpha numeric coordinate, "A6".
Figure 2 illustrates a side view of a modified well
plate apparatus (2) for use in preparation of the library
of the present invention. Well plate (3) contains wells
(4) with a filter (5), and a retaining frit (6), and a
liquid reaction medium used un carrying out the process
(7). The wells have an outlet at the bottom which is
sealed by gasket (8) held in place by a top cover (9) and
bottom cover (10) maintained in position by clamps (11).
Such well plates are typically prepared using
conventional 96-well plates. A hole is drilled in the
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bottom of each well in the plates and a porous frit is
placed in the bottom of each well. The plate is then
placed in the clamp assembly to seal the bottom of the
wells.
Synthesis is initiated by adding reagents to their
individual wells according to their assigned plate
coordinates. The plate is then capped and tumbled to mix
the reagents. Following completion of the reaction, the
solvent and residual volatile reagents may be evaporated
with a Speed-vac. The residual products are redissolved
in appropriate liquid solvent and the reaction products
analyzed, for example, by thin layer chromatography, mass
spectrometry and/or nuclear magnetic resonance
spectrometry.
One embodiment of the present invention is an assay
kit for the identification of pharmaceutical lead
compounds. The assay kit comprises as essential parts,
(1) a well plate apparatus (containing one of the
tetrahydroquinoline compounds in each of its individual
wells), and (2) biological assay materials. The
biological assay materials are generally known to be
predictive of success for an associated disease state.
Illustrative of biological assay materials useful in the
kit of this invention include, but are not intended to be
limited to, those for conducting various assays such as:
In vi tro assays
Enzymatic inhibition,
Receptor-ligand binding,
Protein-Protein interaction,
Protein-DNA interaction,
and the like;
Cell based, functional assays:
Transcriptional regulation,
Signal transduction/Second messenger,
Viral Infectivity,
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Bacteriocidal/Bacteriostatic,
Fungicidal/fungistatic,,
and the like; and
Add, Incubate, & Read assays:
Scintillation Proximit~~ Assays,
Glucan Synthase (GS) inhibition assays,
Angiotensin II IPA receptor binding assay,
Endothelia converting Enzyme [125I] SPA assay,
HIV proteinase [125I] ;;pA enzyme assay,
Cholesteryl ester transfer (CETP) [3H] SPA assay,
Fluorescence Polarization Assays,
Fluorescence Correlation Spectroscopy,
Calorimetric biosensors,
Ca2+ -EGTA Dyes for Cell-based assays,
Receptor Gene Constructs for cell based assays,
Luciferase, green fluorescent protein, beta-
lactamase,
Electrical cell impedance sensor assays, and the
like,
and the like assays.
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Preparation 1
Methods for Constructing ATA compounds
Method A
O O
BocHN BocHN
-~
OH OH
NH2 NHAlloc
22A 23A
BocHN
OMs ---- BocHN
NHAlloc
~OH
NHAlloc
25 A
24 A
BocH ~H BocHN ~
~~~OH
NHAIIoc O NHFmoc O
26 27
N-oc-Fmoc-N-E-Boc-L-Lvs-f~CH2Sl-Glv (27)
Referring to the Method A Scheme shown above (bold-
face compound numberals correspond to bold-face compound
numberals referred to below), in a one liter 3-necked
25 flask fitted with a magnetic stir bar, 22.65 grams
(92mmo1) of commercially available N-E-tert-
butoxycarbonyl-L-lysine 22A and 11.56 grams of sodium
carbonate (110mmo1) were added to 400m1 of water and
stirred to give a suspension. Addition of 400m1 dioxane
to the suspension produced a clear pale yellow solution
on stirring. This solution was chilled on an ice-water
bath with stirring, then 13.2 grams (11.72m1, 110mmo1) of
allyl chloroformate was added over several hours via a
dropping funnel. Stirring was continued overnight without
replenishing the ice bath to yield a light slurry.
The reaction mixture was basified with a few ml of
1N NaOH, then washed twice with 800m1 portions of ethyl
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acetate (EtOAc). The aqueous layer was acidified to pH ~3
with 1N sodium bisulfate (N;~HS04) whereupon a heavy
slurry formed. The slurry w;~s extracted twice into 800m1
portions of EtOAc, then the organic layers were combined,
washed with saturated brine solution, dried over sodium
sulfate (Na2S04) and concenl:rated in vacuo to produce
35grams (>100%) of 23A, a viscous pale yellow oil. TLC
analysis (50:50:1 hexanes:Et~OAc:Acetic acid) showed clean
product and this was carried to the next step assuming
quantitative yield. The crude mass of 23A was dissolved
in 150m1 ethylene glycol dirnethyl ether (DME), chilled to
-15°C (ice/water/NaCl slurry) with stirring, then 9.3
grams (10.11m1) of N-methylmorpholine (NMM) was added and
the solution re-chilled. Isobutyryl chloroformate (12.56
grams, 11.93m1) was then added in one portion to produce
a heavy white precipitate and the resulting slurry was
stirred vigorously for 10 minutes on the brine/ice bath.
The slurry was filtered through a fritted funnel into a
two liter erlenmeyer flask, retentate washed 5 times with
25m1 portions of DME arid thE~ combined filtrate and rinses
were re-chilled to -15°C prior to portion-wise addition
of a solution 5.22 grams (138mmo1, l.5eq) sodium
borohydride (NaBH4) in 150m1 water. Vigorous evolution of
gas was observed after each portion of NaBH4 was added
and the mixture was stirred for an additional 10 minutes
on the ice/brine bath to produce a colorless emulsion.
TLC (3:1 CH2C12:Et0Ac) revea:Led good conversion to the
alcohol. The emulsion was made basic with a few ml 1N
NaOH, then extracted three times with 600m1 portions of
EtOAc. The organic layers were combined, washed with
saturated brine, then dried over Na2S04 and concentrated
in vacuo to yield 25 grams (860) of crude product 24A as
a pale yellow oil.
The product was purified on a 500m1 silica column
packed and eluted with 7:3 C:H2C12:EtOAc to afford 17.5
grams of a nearly colorless ~~il 24A, 60o overall isolated
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yield for alloc protection and acid reduction steps. In a
250m1 roundbottom flask, a portion of pure 24A (7.65
grams, 24mmo1) was dissolved in 100m1 dry THF under a
nitrogen atmosphere, chilled on an ice bath then 5.06m1
triethylamine (3.67 grams, 36mmo1) was added via syringe.
The solution was stirred for a few minutes on ice, then
2.06m1 (3.05 grams, 26mmo1) of methanesulfonyl chloride
(MsCl) was added over a 10 minute period. The clear
solution immediately turned cloudy on addition of the
MsCl and became a heavy white suspension after all of the
MsCl had been added. Stirring was continued for 10-15
minutes following MsCl addition when TLC (3:1
CH2C12:EtOAc) indicated complete consumption of starting
alcohol. The slurry was filtered, retentate washed 3
times with 100m1 portions of EtOAc, then the combined
filtrate and washes were washed once with cold water,
once with cold dilute hydrochloric acid, once with
saturated sodium bicarbonate (NaHC03) and once with
saturated brine before drying over Na2SOg and
concentration in vacuo High vacuum drying of the residue
produced 9.22 grams (96%) of 25A as a colorless oil.
In a 250m1 flame-dried flask fitted with a magnetic
stir bar, 1.28 grams (0.97m1) of mercaptoacetic acid was
dissolved in 30m1 of dry dimethylformamide (DMF) under
nitrogen atmosphere and to the mercapto acid was added
via syringe 6.4m1 (1.5 grams, 2eq) of a 25 weight percent
solution of sodium methoxide in methanol (NaOMe/MeOH).
The clear acid solution turned cloudy white during
NaOMe/MeOH addition then returned to its clear and
colorless appearance within 30 seconds after the addition
was complete. 5 grams of mesylate 25A dissolved in 15m1
dry DMF was then added by pipette in one portion to the
stirring dibasic acid and the resulting mixture stirred
overnight at room temperature under a blanket of
nitrogen. After ~15 hours of stirring, the mixture turned
to a heavy slurry with a gray-pinkish color. TLC (50:50:1
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hexanes:EtOAc:acetic acid) ahowed good conversion, so
most DMF was removed with a rotovap at 45°C bath
temperature to yield a grayish-white paste. The paste was
dissolved in 200m1 water wii:h a few ml of 1N NaOH, and
the resulting turbid solution was washed 3 times with
150m1 portions of ethyl ethE~r. The partially clarified
aqueous layer was then acidified to pH ~3 with NaHS04 to
yield a heavy white slurry that was extracted 3 times
with 200m1 portions of EtOAc. The combined EtOAc extracts
were washed with saturated brine, dried over Na2S04, then
concentrated to afford 4.9 crrams of 26 (98%) as an oil.
The crude product looked relatively clean via TLC
(50:50:1 hexanes:EtOAc:acetic acid) and was carried on
without purification.
To 5.1 grams of 26 under nitrogen in a 250m1 flame-
dried flask fitted with stir bar was added 60m1 dzy THF
via cannula and the mixture stirred to give a clear and
colorless solution. The septum was then removed and in
succession were added 3.36 grams solid 5,5-dimethyl-1,3-
cyclohexanedione (dimedone), 787 milligrams (0.25eq)of
solid triphenylphosphine (PPh3) and 693 milligrams
(0.05eq) of solid
tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4) to
produce a clear orange solution. The flask was recapped,
covered with aluminum foil and flushed with nitrogen
while stirring at room temperature. TLC in 50:50:1
hexanes:EtOAc:acetic acid showed complete conversion of
starting material to a baseline spot after about 1 hour
of stirring. Solvent was removed with a rotovap to give
an orange oil that was then redissolved in 50m1 EtOAc and
extracted 4 times with 25m1 portions of water. To the
combined aqueous layers was t:hen added 2 grams solid
sodium carbonate and 100m1 of. dioxane.
The resulting light emu7.sion was chilled on an ice
bath and when the temperature' of the solution was below
10°C, 4.46 grams of solid fluorenylmethyloxycarbonyl
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chloride (Fmoc-Cl) was added portion-wise over a one hour
period with stirring. The reaction mixture was then
stirred overnight without replenishing the ice bath and
TLC indicated good conversion of starting baseline
material. Next added a few drops of 1N NaOH to the
mixture and washed 3 times with 200m1 portions of ethyl
ether. The aqueous layer was then acidified to pH ~2 with
1N NaHS04 and the off-white slurry was extracted 3 times
with 150m1 portions of EtOAc. The EtOAc layers were
combined, washed with saturated brine, dried over Na2S04,
then concentrated in vacuo to provide a yellow oil
contaminated with dimedone. The oil was applied to a
500m1 silica gel column packed with 50:50:1
hexanes:EtOAc:acetic acid and then eluted with the same
solvent. Appropriate fractions were pooled, concentrated,
then co-evaporated 3 times with toluene/EtOAc to
azeotropically remove excess acetic acid. The resulting
colorless oil was extensively dried under high vacuum to
yield 2.2 grams (35%) of pure 27.
Method B
OH ~OH OMs
-' ' J
H Boc Boc
28 2g 30
32 ~ ~ ~ 1
moc O OH Boy O OH
Referring to the Method B Scheme shown above (bold-
face compound numerals correspond to bold-face compound
numerals referred to below), in a one liter 3-necked
flask, 11.51 grams (100mmo1) of ~3-piperidinemethanol
(28) and 10.6 grams (100mmo1) sodium carbonate were
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dissolved in 200m1 of water, then 200m1 of dioxane was
added and the mixture stirred to produce a pale yellow
light emulsion. To the stirring emulsion was next added
24 grams (110mmo1) of di-tert-butyl-dicarbonate and
stirring continued overnight. TLC (75:25 CH2C12:EtOAc)
revealed complete conversion, so the light yellow slurry
was stripped of dioxane, then extracted twice with 300m1
portions of EtOAc. The organic layers were combined,
washed with saturated brine, dried over Na2S04, then
concentrated in vacuo to yield 20.6 grams (95~) of clean
29 as an off-white solid.
To 20 grams (93mmo1) of 29 under nitrogen in a
flame-dried one liter flask equipped with a stir bar was
added 400m1 dry THF via cannula. The resulting pale
yellow-tan solution was chilled on an ice-water bath,
then 14.12 grams (19.44m1, 1:39mmo1) of triethylamine was
added by pipette and the solution stirred for a few
minutes under nitrogen. Next, 11.72 grams (7.92m1,
102mmo1) of methanesulfonyl chloride was added by syringe
over a 10 minute period with vigorous stirring. The clear
solution turned to a cloudy whitish-tan suspension
immediately upon mesylate addition and became a heavy
slurry within a few minutes after all methanesulfonyl
chloride had been added. TLC (3:1 CH2C12:EtOAc) revealed
complete conversion of the product within 10 minutes of
final sulfonyl chloride addition. The slurry was filtered
through a fritted funnel and the retentate rinsed 4 times
with 100m1 portions of EtOAc. The combined filtrate and
rinses were then washed once each with cold water, cold
dilute hydrochloric acid, saturated NaHC03, and saturated
brine. The organic layer was dried over Na2S04 and
concentrated in vacuo to provide 26.94 grams (980) of 30
as a pale yellow crystalline solid. ,
2-Mercaptonicotinic acid (11 grams, 7lmmol) was
dissolved under nitrogen in 400m1 dry DMF under nitrogen
in a one liter 3-necked flask. fitted with a large egg-
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shaped stir bar to give a bright yellow solution. Next,
7.67 grams (32.5m1, 142mmo1) of a 25 weight per cent
solution of NaOMe in MeOH was added to the stirring
mercaptonicotinic acid/DMF solution, then 20.8 grams
(71mmo1) of 30 dissolved in DMF (52m1 final volume) was
added by syringe pump over a two day period. The
resulting deep yellow viscous slurry was stripped of DMF,
then redissolved in 400m1 of water. The turbid yellow
aqueous solution was partially clarified by 3 washes with
300m1 portions of ethyl ether. The partially clarified
aqueous layer was acidified to pH ~2 with 1N NaH504 to
produce a heavy yellow slurry that was then extracted 3
times with 300m1 portions of EtOAc. The organic layers
were combined, washed with saturated brine, dried over
Na2S04, and concentrated in va uo to provide 24.6 grams
(98%) of crude 31 as a canary yellow solid.
The crude product was recrystallized from neat EtOAc
to provide two crops of pale yellow crystals weighing
20.95 grams (83o isolated yield). Purified acid 31 (19
grams, 54mmo1) was combined with 108m1 of a 4N
hydrochloric acid solution in dioxane and stirred in a
one liter flask to produce a heavy white-caked
precipitate. 100m1 of dioxane was added to loosen the
slurry and after one hour of stirring, TLC (50:50:1
hexanes:EtOAc:acetic acid) indicated complete conversion
of starting material to a baseline product. A large part
of the HC1 was removed under light vacuum on a rotovap,
then the remains were neutralized with 1N NaOH solution.
To the neutralized solution was then added with stirring
250m1 of water, 5.72 grams (54mmo1) of solid sodium
carbonate and another 100m1 of dioxane. The resulting
emulsion was chilled on an ice-water bath, then 19.12
grams (56.7mmo1) of fluorenylmethyloxycarbonyl N-
hydroxysuccinimide (Fmoc-OSu) was added portion-wise over
a 10 minute period. Stirring was continued for 36 hours
to produce an ivory slurry. Dioxane was removed in vacuo,
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then the mixture was washed 3 times with 300m1 portions
of ethyl ether. The aqueous layer was then acidified to
pH ~2 with 1N NaHS04 and the resulting heavy slurry
extracted 3 times with 300m1 portions of EtOAc. The
combined EtOAc layers were w~~shed with saturated brine,
dried over Na2S04, then concentrated and dried under high
vacuum to yield 23.54 grams (92%) of 32 as pale yellow
solid that was judged clean by TLC in 50:50:1
hexanes:EtOAc:acetic acid.
Method C
O
Bn0 OOH --- BnO~OH ---~ BnO~OMs
oc NHB ,
NH Boc
33 34 35
Me Me
BnO~S OH OH
'-- BnO~S
NHFmoc O NHBoc O
37 36
N-a-Fmoc-O-benzvl-L-Ser-ft-CH~S1-DL-Alanine !37>
Referring to the Method C Scheme shown above (bold-
face compound numerals correspond to bold-face compound
numerals referred to below), in a 500m1 round-bottomed
flask fitted with a magnetic stir bar, 25 grams (84.6
mmol) of N-oc-Bac-O-benzyl-L-Serine (33) (Novabiochem, San
Diego, CA, USA) was dissolved in 150 ml DME and chilled
to -15°C with an ice/water/brine bath. 10.23 ml N-
methylmorpholine (93 mmol, 1.1 eq) was added to the
chilled solution, stirred a few minutes, then 12 ml of
isobutyl chloroformate (93 mmol, 1.1 eq) was added in one
portion at which time the solution quickly turned to a
heavy white slurry. The slurry was stirred vigorously
for 10 minutes on the brine/i~e bath, then filtered
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through a fritted funnel into a two liter erlenmeyer
flask. The retentate was washed 5 times with 15 ml
portions of DME and the combined filtrate and rinses were
re-chilled to -15°C prior to portion-wise addition of a
solution 5.3 grams (140mmo1, 1.66eq) sodium borohydride
(NaBH4) in 100 ml water. Vigorous evolution of gas was
observed after each portion of NaBH4 was added. The
mixture was stirred for an additional 10 minutes on the
ice/brine bath to produce a colorless emulsion. TLC (3:1
CH2C12:EtOAc) revealed good conversion to the alcohol.
Reaction was quenched with 2000 ml of water and the
resulting emulsion was made basic with a few ml 1N NaOH,
then extracted three times with 300m1 portions of EtOAc.
The organic layers were combined, washed with saturated
brine, then dried over Na2S04 and concentrated in vacuo to
yield a pale yellow oil. Product recrystallized from 400
ml of 7:1 petroleum ether: diethyl ether to give 20.28
grams (85%) of alcohol 34 as white needles after washing
with hexanes and drying under high vacuum.
In a 1000m1 round-bottomed flask, a portion of pure
34 (9.0 grams, 32 mmol) was dissolved in 125 ml dry THF
under a nitrogen atmosphere, chilled on an ice bath, then
6.69 ml triethylamine (4.86 grams, 48 mmol) was added via
syringe. The solution was stirred for a few minutes on
ice, then 2.73 ml (4.03 grams, 35.2 mmol) of
methanesulfonyl chloride (MsCl) was added over a 10
minute period. The clear solution immediately turned
cloudy on addition of the MsCl and became a heavy white
suspension after all of the MsCl had been added. Stirring
was continued for 10-15 minutes following MsCI addition
when TLC (3:1 CH2C12:Et0Ac) indicated complete consumption
of starting alcohol. The slurry was filtered, retentate
washed 3 times with 100 ml portions of EtOAc, then the
combined filtrate and washes were washed once with cold
water, once with cold dilute hydrochloric acid, once with
saturated sodium bicarbonate (NaHC03) and once with
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saturated brine before drying over Na2S04 and
concentration in vacuo. High vacuum drying of the residue
produced 12.7 grams (>100%) of 35 as a colorless oil.
In a 250m1 flame-dried flask fitted with a magnetic
stir bar, 1.91 grams (1.6 ml) of DL-thiolactic acid was
dissolved in 40 ml of dry di:methylformamide (DMF) under
nitrogen atmosphere and to t:he mercapto acid was added
via syringe 8.24 ml (1.94 gr,~ms, 2eq) of a 25 weight
percent solution of sodium m~~thoxide in methanol
(NaOMe/MeOH). About 5.4 grams of mesylate 35 (~15 mmol)
dissolved in 35 ml dry DMF w~~s then added by pipette in
one portion to the stirring dibasic acid and the
resulting mixture stirred ovc=rnight at room temperature
under a blanket of nitrogen. After ~15 hours of stirring,
the mixture turned to a heav~r slurry with a grayish
color. TLC (50:50:1 hexanes:EtOAc:acetic acid) showed
good conversion, so most DMF was removed with a rotovap
at 45°C bath temperature to ~~ield a grayish-white paste.
The paste was dissolved in 200m1 water with a few ml of
1N NaOH, and the resulting turbid solution was washed 3
times with 150m1 portions of ethyl ether. The partially
clarified aqueous layer was then acidified to pH ~3 with
NaHS04 to yield a heavy whites slurry that was extracted 3
times with 200m1 portions of EtOAc. The combined EtOAc
extracts were washed with saturated brine, dried over
Na2SOq, then concentrated to afford 5.6 grams of 36 (101%)
as an oil. The crude product looked relatively clean via
TLC (50:50:1 hexanes:EtOAc:acetic acid) and was carried
on without purification. 3.8 grams of 36 (10.4 mmol) in a
1000 ml round bottomed-flask was chilled on an ice-water
bath and combined with 100 ml of a 4N hydrochloric acid
solution in ethyl acetate. TLC (50:50:1
hexanes:EtOAc:acet:ic acid) indicated complete conversion
of starting material to a baseline product within one
hour of HC1 addition. Most of the HC1 was removed under
vacuum on a rotovap, then the remains were dissolved in
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100 ml of water. To this solution was then added with
stirring 4.4 grams (42 mmol, 4 eq) of solid sodium
carbonate and then 75 ml of dioxane. The resulting
emulsion was chilled on an ice-water bath, then 3.86
grams (11.4 mmol) of fluorenylmethyloxycarbonyl N-
hydroxysuccinimide (Fmoc-OSu) was added portion-wise over
a 10 minute period. Stirring was continued overnight and
TLC (50:50:1 hexanes:EtOAc:acetic acid) of the resulting
ivory slurry showed complete consumption of starting 36.
Dioxane was removed in vacuo, the mixture was diluted
with 200 ml and then washed 3 times with 200m1 portions
of ethyl ether. The aqueous layer was then acidified to
pH ~2 with 1N NaHS04 and the resulting slurry extracted 3
times with 100m1 portions of EtOAc. The combined EtOAc
layers were washed with saturated brine, dried over
Na2S04, then concentrated and dried under high vacuum to
yield 5.6 grams (110%) of 37 as a colorless oil.
Recrystallization from acetone-hexanes failed, but TLC
(50:50:1 hexanes:EtOAc:acetic acid) indicated ~80%
purity.
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Prebaration 2
The following ATA compounds have been prepared in
accordance with the general methods of Scheme II and can
be utilized to form the macrocycle library of the present
invention.
O OH N/
OH \
/ S OH ~'' I \ S
\ I NHFmoc NHFmoc O / NHFmoc
O OH
FmocH
/ iN
/S OH \ I N
S S OH
NHFmoc O / p OH NHFmoc
O O
~~~ ~ OH ~~
~S~OH ~S~ S~~~OH
NHFmoc " NHFmoc
HFm~c ~ NH~II~c
/ /
O
~ ~ \ I S \ I OH
S' Y 'OH S
O
NHFmoc NHBoc NHFmoc N
O OH Fmoc
N/
O
BocH OH ~ ~ ~ S \
S :> Y 'OH
NHFmoc
NHFmoc O WHFmoc NHBoc O OH
NHBoc
/ I NHFmoc S OH
Bn~S OH \ \S \
S O
NHFmoc O I / NHI=moc / ~ I \
O OH \ /
OH ~ OH
_S ~ wS
NHFmoc O NHFmoc p
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BocH OH
NHFmoc O
OH
S
NHFmoc O
O OH
'S
O NHFmoc O
O
OH
CbzH OH
S
NHBoc O
H
OH
~S
O NHFmoc O
0 S~OH O OH
S
NHFmoc O , NHFmoc O
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OH
~S OH
S
Fmoc O
NHFmoc O
O
~OH
OH
'S OH
S
Fmoc O
NHFmoc O
O
~S OH S OH
NHFmoc
O NHFmoc
O
S~ OH
F
H moc
O
~S OH S OH
NHFmoc , NHFmoc p
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O
~S ~ ~OH
NHFmoc NHBoc
O
~S ~ ~OH
NHFmoc NHBoc
_ OH
S ~ ~S
NHFmoc O
OH
~S
NHFmoc O
OH
~S ~ ~S ~ S OH
NHFmoc 0
NHFmoc 0 ,
O
S OH Si~OH S OH
NHFmoc O , NHFmoc , NHFmoc O
S OH S~ /OH
NHFmoc O , NHFmoc ~O
O'~S~OH
NHFmoc IIO
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-109-
O~S OH S \
NTHFmoc O NHFmoc
O OH
O O
\~~S OH ~S OH
NHFmoc NHBoc , NHFmoc NHBoc
O
/~S OH
NHFmoc NHBoc
O OH
S \
\ ~' ~S
FmocN / ~ /- NHFmoc
O' ~OH
O
J
OH ~ S
FmocN NHI=moc
O OH
N/ O
s
'S OH
Fmoc
O OH F~nocN
\ ~ OH ~O S \
\~S
NHFmoc
NHFmoc O O OH
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-110-
'OH
I ~II(S
O
N
Fmoc
S \ S \
NHFmoc NHFmoc
O OH O OH
N~
~S
F oc O ~ ~OH
S \
/OH
I ~S
NHFmoc O ~ moc O OH '
H
\ OH
\ S
J
Fmoc O
O OH , Fmoc
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-:L11-
N~N~N O
S \ OH ~ N\ ~
S~ v 'OH
NHFmoc O
NHFmoc
O O
S~~~OH
~S = OH
NJ ~ Fmoc NHAlloc
Fmoc
N~N~N O
~--~N\ ~
S- v 'OH
NHFmoc Fmoc
H
FmocHl
OH
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-112-
FmocHN
and
NHBoc
FmocHN S OH
/ S \ O
\ ~ ~ /
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-113-
Example 1
Method for Preparation of Libraries of Macrocvclic
Pseudopeptides and Oraanopeptides
Trt O
Trt O N
FmocHN~ ~ N'~~ _~ NH H
FmocHN H
'O -
6 FmocH ~~ ~ ~ Otbu
7
c
Trt p
N'HFmoc N
mr~
~~u H
of
H
Cvclo - (Cvs-Tvr-2-Nic fy~H,~.,;;~ -Phe-D-Ala-a-acetate-
CH Sl)-(11)
100mg (50micromoles) of Fmoc-Rink-Amide AM or Fmoc-
Rink-Amide MBHA resin was placed in a polypropylene
reaction vessel (one of 36 such vessels contained in the
reaction block of an Advanced Chemtech Model 357 multiple
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-114-
peptide synthesizer). Referring to Scheme III above
(bold-face compound numerals and lettered steps
correspond to bold-face numerals and lettered steps
referred to below), steps (a) through (e) were performed
robotically at room temperature according to a custom
program. The resin was then washed twice with 1.5m1 of
N-methyl-pyrrolidinone (NMP). In Step (a), the Fmoc
protecting group was removed by agitating the resin with
1 ml of 30o piperidine in dimethylformamide (DMF) for 3
minutes. The resin was drained and a second 1ml portion
of piperidine/DMF solution was added, agitated for 10
minutes, then drained. The resin was then washed and
drained 5 to 6 times with 1.5m1 portions of NMP. To the
washed resin was then added 300uL of NMP followed by 1m1
of a 0.5M solution of N-Fmoc-S-trityl-L-Cysteine (5
equivalents of protected amino acid to resin-bound amine)
and 0.5M hydroxybenzotriazole (HOBT) in NMP and 250u1 of
a 1.OM solution of diisopropylcarbodiimide (DIC) in NMP.
This mixture was then agitated for 45 min., drained and
the resin rinsed 4 times with 1m1 portions of NMp.
The coupling and rinse steps were repeated one time
to yield resin-bound intermediate 6. Single coupling was
also acceptable in many cases. Intermediate 6 was then
subjected piperidine, NMP washes and coupling as for step
(a), but with a 0.5M solution of N-Fmoc-L-Tyrosine-O-
tert-butyl ether and 0.5M HOBT in NMP instead of the
protected cysteine derivative to produce resin-bound
intermediate 7. In step (c), 7 was subjected to the
piperidine, washes and coupling regimen of steps (a) and
(b), but with a 0.5M solution of N-Fmoc-L-phenylalanyl-
[~lCH2S]-2-mercaptonicotinate and 0.5M HOBT in NMP to give
compound 8
Compound 9 was produced in step (d) by subjecting
compound 8 to the piperidine wash and coupling conditions
outlined in step (a) but with 0.5M N-Fmoc-D-Alanine and
0.5M HOBT in NMP. Terminal alpha-bromoamide 10 was made
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-115-
in step (e) by treating compound 9 with piperidine/DMF,
washing as in step (a), and coupling with a 0.5M solution
of Oc-bromoacetic acid in NMF~ without added HOBT. HOBT is
sufficiently nucleophilic to displace the bromine and
should thus be omitted when coupling bromoacids.
After coupling of the terminal bromoacid, the resin
was washed 4 times with 1.5m1 portions of NMP, then 5
times with 1.5m1 portions of methanol. After methanol
washes were complete, resin containing attached compound
10 was dried at room temperature under high vacuum for at
least 3 hours. The dried resin was transferred to a
screw-capped vial and cleavage effected by agitating the
resin with 2.5m1 cleavage cocktail (38:1:1
trifluoroacetic acid(TFA):H2O:triethylsilane (Et3SiH) or
19:1 TFA:H20) for 1.5 hours ~~t room temperature. Another
2 ml of cleavage cocktail wa;~ added and agitated for 1.5
hours, then the entire mixture was filtered through a
porous plastic frit. The retentate was washed twice with
2.5 ml portions of water and the combined filtrate and
washes were frozen and lyophilized to yield a yellow-
orange amorphous solid. This solid was mixed vigorously
with 20m1 diethyl ether, chilled to -20°C and centrifuged
at 15K rpm, 4°C for 10-15 minutes, then the ether was
decanted. The remaining off--white pellet subjected to
another ether wash and centr~_fugation run. Ether was
decanted and the pellet allowed to air dry, at which time
the white to off-white solid was dissolved in 10m1 1:1
CH3CN:H20, filtered through a 0.45uM teflon filter disc,
then lyophilized from a tareci scintillation vial. The
crude material was then cycli.zed by dissolution in 10m1
1:1 CH3CN:H20 plus 100uL of 1M DIPEA in CH3CN.
Other bases that promotE~d efficient cyclization were
2,6-lutidine and proton sponcre ([1,8-
Bis(dimethylamino)naphthalene]). Triethylamine, pyridine
and tetramethylguanidine were sufficiently nucleophilic
to displace the bromine in preference to promoting
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-116-
cyclization. Cyclization was allowed to proceed for 24
hours at room temperature. LCMS analysis showed a single
major peak (purity~80o) having an MH+ of 502.2,
consistent with the calculated mass far 11 of 501.4.
The abbreviations "Cys" and "BrA" represent cysteine
and bromoacetic acid residues, respectively, and these
two residues define the ligation point for ring closure.
Although the majority of cycles synthesized to date use
L-Cysteine for ring closure, a number of other cycles
have been successfully constructed with D-Cysteine. In
addition, it is anticipated that other trifunctional
compounds bearing an amine, an acid and a mercapto
function (for example, penicillamine), are likely to
function in the scheme as does the cysteine residue.
Also, other bromoacids will likely function in place of
bromoacetic acid.
Macrocycles 38 and 39 below have been constructed
and demonstrate ATA compatibility with commercially
available amino acids. Further, macrocycle 38 shows that
ATAs provide easy access to novel macrocycles containing
aryl backbone elements.
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-117-
H
O
O 0
a
S
0 0
NH 'Nh~
~O
O
OH
38 39
Example 2
Methods for Construc:_tina Tailed Macrocvclic
Pseudopeotides and Oraanopeotides
According to a general method for constructing
resins bearing both free amine groups) and an acid
labile carbamate linkage, described and shown below,
11 grams (8.58 millimoles) of p-nitrophenyl carbonate
Wang resin (~~pNP-Wang" a commercially available
material from Novabiochem, Catalog# 01-64-0123) was
swollen with 75m1 of 1:1 dichloromethane (DCM or
CH2C12:N-methylpyrrolidinone (NMP), then combined with
a 100m1 solution containing 429mmo1s (5 equivalents) of
the diamine of choice. The resulting bright yellow
slurry is then agitated on a peptide shaker, rotor or
other mixing device for 24-48 hours at room
temperature. The slurry is next drained using a fritted
funnel or peptide shaker flask, then washed with 200m1
each of DCM, NMP and methanol. The wash cycle (DCM, NMP
and methanol) is repeated i:wice more, then the resin is
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-118-
dried under high vacuum for several days to provide the
desired product in quantitative yield based upon mass
change.
By this method was prepared piperazine-N-carbamoyl-Wang
(resin P) (loading: 0.81mmo1/gram of resin) as well as
both p- and m-xylenediamine-N-carbamoyl-Wang resins
(resins PX and MX, respectively at 0.78mmo1/gram) in
multigram quantities.
Alternatively, the desired diamine (or higher
order amine) can be similarly attached to resin in situ
by mixing at least a five-fold molar excess of di- or
higher order amine with pNP-Wang resin in 1:1 DCM:NMP
for 16-48 hours at room temperature. After washing as
above, the resin contains at least one free amino
function ready for elaboration and at least one acid
labile carbamoylated amine function as the link to
resin.
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-~119-
r
s
a
0
x
0
s ~ c 5 c
w
> w
~i O Q Q Q
a
Q e: o' c c
O ~ ~ ~ -v '° 3 w
a a. 3 3
O a ~ W i4 S. p 'a.
4i E ~ E~
d ~_ ~ p
_Of ul ci X w x U w
Z a eo - p. av ~Z amz
Q
d aii 5d c~ -
wc - _ac co
~c :°. ~« ~E
and ~ ~ c
c : ~ d v
GC C'~ c: c v
tEo C aft G ~ a X m
p es 'o Z 8 d y X '
ao : "' 3 oC c~ a E '~_
c G. o h a C
d a oVr m
a-° ~r.e n
a N o~~ E
w_C o ~
E ~ a to
\ /
dd o
r~ d
NO N o
/ ~ 0 0 0
'~° E
0
v' / /
o v
o-~~ 3
o z o 0 0 0
O O O
z~ a zs z
\ //
z = \ / \
0
x
z z
n n
x x
SUBSTITUTE SHEET (RULE 26)
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WO 98/46631 PCT/US98/07135
-120-
N-r~-xvlenediamino-Cvclo-(Cys-Phe-Glv[t(1CH?S1-Phe-Arg-a
acetate-[~rCHZSJ? (18, Scheme IV)
w
N ~'
U
s < < m
m <
\ zx \ g a
o I o I /
O / Z 'GO :.
N . ~
,
H ~. I~ H
O
zx
x
z ~o v
/ O ~~~ vi < a
x N ~ a
z~ xz_ ,_z o =-~!~ n a
_~ \ <
I\
I / m m
H E
d ~ a
w
t
vd ~ U
ewg ~ \ o a I \ ~
I / .- z o / ~ a
m = o m N //
_ a z z ,.
J
0
z
x xz o
."~ a.
U y C ~ ~ ~<n v a
cn '_ ~ a /z o
U c_si \ a
z I /
o a
t
z ~ w
m
z a \
0
.,.
e.
r O OD ~ U
Q ~ O _~o
__
Z
\ _
_
/
SUBSTITUTE SHEET (RULE 26)
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Steps: a) PX-Wang resin, then 5 eq Fmoc-S-trityl-L-cysteine, DIC/HOBT in NMP,
wash; b) 30% piperidine in DMF, wash, then 5 eq Fmoc-L-phenylalanine; c) as
for b), but with 5 eq of Fmoc-L-Phe[yrCH2S]-Gly-OH; d) as for b), but with 5
eq of
Fmoc-L-Arg(Pmc)-OH where Pmc is defined below; e) as for b), but with 5 eq
bromoacetic acid and DIC in NMP without HOST; f) 38:1:1 TFA:H20:Et3SiH for
24 hr., lyophilize, then 2 eq of diisopropylethylamine in 1:1 CH3CN:H20.
With respect tc Scheme IV (bold-face compound
numerals and lettered step~~ correspond to bold-face
compound numerals and lettered steps referred to below),
65mg (50micromoles) of PX resin was placed in a
polypropylene reaction ves~;el Cone of 36 such vessels
contained in the reaction flock of an Advanced Chemtech
Model 357 multiple peptide synthesizer). Referring to
Scheme IV, steps (a) through (e) were performed
robotically at room temperature according to a cumtom
program written by the bench chemist. The resin was
25
35
SUBSTITUTE SHEET (RULE 26)
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-121-
washed and drained twice with 1.5m1 of N-methyl-
pyrrolidinone (NMP). In Step (a), to washed resin was
then added 300uL of NMP followed by 1m1 of a 0.25M
solution of N-Fmoc-S-trityl-L-Cysteine (5 equivalents
of protected amino acid to resin-bound amine) and 0.25M
hydroxybenzotriazole (HOBT) in NMP and 250u1 of a 1. OM
solution of diisopropylcarbodiimide (DIC) in NMP. This
mixture was then agitated for 45 min., drained and the
resin rinsed 4 times with 1m1 portions of NMP. The
coupling and rinse steps were repeated one time to
yield resin-bound intermediate 13. Single coupling was
also acceptable in many cases. In Step (b), the Fmoc
protecting group of 13 was removed by agitating resin
with 1 ml of 30% piperidine in dimethylformamide (DMF)
for 3 minutes. The resin was drained and a second 1m1
portion of piperidine/DMF solution was added, agitated
for 10 minutes, then drained. Resin was then washed and
drained 5 to 6 times with 2.5m1 portions of NMP.
To washed resin was then added in succession 300uL of
NMP, 1m1 of a solution 0.25M each in N-Fmoc-L-
Phenylalanine and HOBT and 250u1 of a 1.OM solution of
diisopropylcarbodiimide (DIC) in NMP. This mixture was
then agitated for 45 min., drained and the resin rinsed
4 times with 1m1 portions of NMP. The coupling and
rinse steps were repeated one time to yield resin-bound
intermediate 14. In step (c), 14 was subjected to
piperidine, washing and coupling regimen of step (b),
but instead with N-Fmoc-L-phenylalanyl-[ylCH2S]-glycine
to give 15. Compound 16 was produced in step d by
subjecting 15 to the piperidine, wash and coupling
conditions outlined in step (b) but with N-Fmoc-L-
Arginine(Pmc) as the amino acid [where Pmc=2,2,5,7,8-
Pentamethylchroman-6-sulfonyl, an acid labile arginine
side chain protection function]. Terminal alpha-
bromoamide 17 was made in step (e) by treating 16 with
piperidine/DMF, washing as in step (b), and coupling
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-122-
with a 0.25M solution of alpha-bromoacetic acid in NMP
without added HOBT. HOBT is sufficiently nucleophilic
to displace the bromine and should thus be omitted when
coupling bromoacids in this step. After coupling of the
terminal bromoacid, the resin was washed 4 times with
l.5ml portions of NMP, then 5 times with l.5ml portions
of methanol. After methanol washes were complete, resin
containing attached 17 was dried at room temperature
under high vacuum for at least 3 hours. The dried resin
was transferred to a screw-capped vial and cleavage
effected by agitating the resin with 2.5m1 cleavage
cocktail (38:1:2 trifluoroacetic acid(TFA):H20:
triethylsilane (Et3SiH) or 19:1 TFA:H20) for 1.5 hours
at room temperature. Anoi:her 2 ml of cleavage cocktail
was added and agitated for 1.5 hours, then the entire
mixture was filtered through a porous plastic frit. The
retentate was washed twices with 2.5 ml portions of
water and the combined fi=Ltrate and washes were frozen
and lyophilized to yield ~~ yellow amorphous solid.
Workup and cyclization were performed as in Example 1
above. Cyclization was a~_lowed to proceed for 24 hours
at room temperature. LCMS analysis showed a single
major peak (purity~80%) having an MH+ of 790.4 and an
[M+2H]2+ of 395.8, consistent with the calculated mass
for 18 of 789.
The bottom part of the scheme depicts a generic form
of the synthetic method u~;ed to construct macrocycles
such as 18."Cys" and "BrA" represent the cysteine and
bromoacetic acid residues, respectively and these two
residues define the ligation point for ring closure.
Though the majority of cycles synthesized used L-
Cysteine for "Cys", a number of cycles were
successfully constructed with D-Cysteine at the "Cys"
position.
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-123-
Example 3
Following the procedures of Example 1 above, the
following library compounds have been prepared. The
number within the ring specifies the number of ring
atoms.
S O
HN 12 ' NH2
NH
S B n S ~~O
O
-'S
HN O
13
NH NH2
S
O
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WO 98/46631 PCT/US98/07135
-1.24-
HN
,,
1 ~3
NH NH2
~N
14
NH NH2
S
O
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-125-
U
Met
R ~ H N~ R~=
S H
15 °
Me
NH NHt ~
~ Me'
S-r \\
O M ,"
/ Me
\ H~~'~a~~~~"
O Me
~.
H H x~"
S
15 ° H
HN °
O
M~''~~, NH N~
M,~~
S O H C
Me
I
w."
NHZ \ ~,"
4
HO /
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-:126-
O
O R'~ N~ R~ H
p ,~-r' N
" 16 0
N 16
O
HO NH NH 2
S NH NH 2
~O
O
O ~ ~ ; H O
N
S
O S ~ S
16 o S 16 0
~N V N
,NH NH2
NH NHZ
N
O H r';~' O N-T'
R2 O H ~~' O
R2
O Me
R~_ H2N\/\/H / xIIII
y"~ ~ W~ ~ ~S'y~ ~~S'H'~ HO~SSyh y"4
HO~
Me
HO
Me
Me' ' S '~ Met HO
HO~
O
Rr H~\/~\/~'w1 \ ~~b
\~°"~~ HO~~'v, O
HO / O HO~~""~ ~
Me
NH
Me~SW~v H2N~NW~~~ HO/~~'',
H
Me
Me' ' \ \ ~~ HO HO~
/
HO
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-127-
y H O
s
S
O
NH N~
o M8
H Nw,
Rt- 2 ~..~ I \ ~~.y~ I \ '.y~ ~~~.y~ Hp~~.ys HO~sy,
HO
Me
Me' ' \ S ~~ HO HO~
I /
Rx HpNw.,~~ I \ ',~Y \ y, ~ Hp~y~., p ~ ,
'~,
~ /'~a
HO / O HO' v "~
NH
H Me~S~~~~~ Hz~'~~N~~~'~ HO/v,''.
H
Me
Me' ' \ \ ~~ HO HO~
I~ I/
HO
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-~_28-
O
N
R' / H
O S,
O
N 17
NH NH2
~O
~l
N
R~_ H2N "~"~ \ /"w, \ ",
~, ""' "~"" Me~"~""",
HO ~ ~ Me
O Me Me
~ /., ~
HO' v ""'~ "'"w
HO Me' ' \
S ~ Met f.~0 HO~
O
CA 02286867 1999-10-12
WO 98!46631 PCT/US98/07135
-129-
HI
Me~."",~
R
Rte. ~-~ ,~' ~
s
HN
18 ~ Me 18
NH NH2
g ~ S
O OH
O ~ ~ O
O
R2 N~ R
H
18
~-- Me'
I/ H Me~,
O C
Me
H 2N ~/\/""'".", M a "" ~ /""
"x '"'' HO- v """~'
HO °''
Me
''"i~,"
HO
Met H~ Me
HO
Me
HpN ~","w" HO ~~~,~"~ HO/"~,~~
U
0
~~ R~ ~~
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-:L30-
0
H N g'
Me O
19
S NH N~
O
H2N O H \
Me OH
R~= H2N h~wh, M
Me
hAyAyN, \ ~hM~~Yhi
HO / / /
O Me
M "' ~ /~
~~b~ ~~~h~h DIY
HO ~ HO
Me
~ HO~
\ 'OI HO~
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-131-
H O
S
O
N!~
NH
S
i p
H ~ ~ ~ OH
O
Me
R~- H2N ~~'~~~ M ~ H O
M
b w w4
\ .
/ / H / \
O Me
M~/,~,.,~ H
~~~~',
Me O
Rz H O Rz H O
O ~'''~ N ~ N
S O
O \~ S O
N
NH NH z Z 1 NH NH z
O S
N ~O
t
H ~ ~ OH ~ ~ H OH
O
R 2. H zN w~~,~ H O 'ww i"w
HO '~
O
