Note: Descriptions are shown in the official language in which they were submitted.
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1
PIPERAZINE DERIVATIVES FOR THE TREATMENT OF HIV INFECTIONS
This invention relates to piperazine derivatives, to processes for their
preparation, to compositions containing them and to their use.
More particularly, the present invention relates to the use of piperazine
derivatives in the treatment of HIV, such as HIV-1, and genetically related
retroviral infections (and the resulting acquired immune deficiency syndrome,
AIDS).
Entry of HIV-1 into a target cell requires cell-surface CD4 and
1o additional host cell cofactors. It is recognised that for efficient entry
into
target cells, human immunodeficiency viruses require a chemokine receptor,
such as CCR5 or CXCR-4, as well as the primary receptor CD4. The principal
cofactor for the entry mediated by the envelope glycoproteins of primary
macrophage-trophic strains of HIV-1 is CCRS, a receptor for the ~3
t5 chemokines RANTES, MiP-1a and MIP-1~i (Deng et al., 1996, Nature, 38,
661-666). HIV attaches to the CD4 molecules on target cells through a region
of its envelope protein, gp120. It is believed that the CD4 binding site on
the
gp120 of HIV interacts with the CD4 molecule on the target cell surface and
undergoes conformational changes, which allow it to bind to further cell-
2o surface receptors such as CCR5 or CXCR-4. This brings the viral envelope in
closer proximity to the cell surface and allows interaction between gp41 on
the viral envelope and a fusion domain on the host cell surface, subsequent
fusion with the cell membrane and, ultimately, entry of the viral core into
the
cell. Accordingly, compounds that inhibit the binding of gp120 with CD4, and
25 hence prevent the entry of HIV-1 into a target cell, should be useful in
the
treatment of HIV, such as HIV-1, and genetically related retroviral infections
(and the resulting acquired immune deficiency syndrome, AIDS).
According to a first aspect of the present invention, there is provided a
compound of formula (I)
O
R3
R4 N- _R'
5
R~~ N
Rz
o (I)
or a pharmaceutically acceptable salt, solvate or derivative thereof, wherein:
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R' is phenyl or pyridyl, wherein said phenyl or pyridyl is optionally
substituted
by 1 or 2 atoms or groups selected from halo, C1-C6 alkoxy, CF3, OCF3, or
CN;
R2 and R3 are independently H, or C1-C6alkyl;
R4 is C1-C6 alkyl;
1o R5 is phenyl; naphthyl; or a C-linked, 6 to 10 membered, mono- or bicyclic,
aromatic or partially saturated, heterocycle wherein said heterocycle contains
1 to 4 nitrogen heteroatom(s), 1 or 2 nitrogen and 1 oxygen heteroatoms, or 1
or 2 nitrogen and 1 sulphur heteroatoms; wherein said phenyl, napthyl or
heterocycle is optionally substituted by 1 to 3 atoms or groups selected from
C1-C6 alkyl, C1-C6 fluoroalkyl, C3-C~ cycloalkyl, phenyl, OH, C1-Cg alkoxy,
C1-C6 alkoxy C1-C6 alkyl, OC1-C6fluoroalkyl, Co-C2 alkylene NR6R', halo,
Co-C2 alkylene CN, Co-C2 alkylene C02R8, Co-C2 alkylene CONR6R',
Co-C2 alkylene SR9, Co-C2 alkylene SOR9, Co-C2 alkylene S02R9,
Co-C2 alkylene SO2NR6R', Co-C2 alkylene NR$CORg,
2o Co-C2 alkylene NR$CONR6R', Co-C2 alkylene NR8SO2R9, or
Co-C2 alkylene R'°, or, where R5 is a heterocycle, oxo;
R6 and R' are independently H, C1-C6 alkyl, C3-C~ cycloalkyl, phenyl or Rlo;
or when taken together with the nitrogen to which they are attached form an
optionally substituted azetidine, pyrrolidine, piperidine, morpholine, or
thiomorpholine ring; wherein the said substituents are 1 or 2 groups selected
from C1-C6 alkyl, or Co-C° alkylene NH2;
R$ is H, Ci-C6 alkyl or phenyl;
R9 is C1-C6 alkyl or phenyl; and
R'° is imidazolyl, pyrazolyl, triazolyl, thienyl, furyl, thiazolyl,
oxazolyl,
thiadiazolyl, oxadiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl,
quinolinyl, isoquinolinyl, benzimidazolyl, indazolyl, quinazolinyl,
phthalazinyl,
benzoxazolyl or quinoxalinyl, each optionally substituted by 1 to 3 atoms or
groups selected from C1-C6 alkyl, C1-C6 alkoxy, cyano or halo.
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The term "alkyl" as a group or part of a group includes straight chain
and branched groups. Examples of alkyl include methyl, ethyl, n-propyl,
i-propyl, n=butyl, i-butyl, sec-butyl and t-butyl. The term "C3_7 cycloalkyl"
means cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. The
term "halo" means fluoro, chloro, bromo or iodo.
In a further embodiment, R' is phenyl or pyridyl, wherein said phenyl or
pyridyl is optionally substituted by 1 or 2 atoms or groups selected from
halo.
In yet a further embodiment, R' is phenyl, fluorophenyl or pyridyl.
In yet a further embodiment, R' is phenyl.
o In yet a further embodiment, R2 is C~-C4alkyl.
In yet a further embodiment, R2 is methyl.
In yet a further embodiment, R3 is H.
In yet a further embodiment, R4 is C1-C4alkyl.
In yet a further embodiment, R4 is methyl.
In yet a further embodiment, R5 is an optionally substituted phenyl,
naphthyl, pyridyl, indazolyl, benzimidazolyl, quinolinyl, isoquinolinyl,
quinazolinyl, benzopiperidinyl or benzoxazolyl; wherein said substituents are
1
to 3 atoms or groups selected from C1-C6 alkyl, C~-C6 alkoxy, halo, CN,
CO2R8, CONR6R7, or R'o.
2o In yet a further embodiment, R5 is an optionally substituted phenyl or
pyridyl, wherein said substituents are 1 to 3 groups selected from C1-C6
alkoxy, C02R8, or CONR6R'.
In yet a further embodiment, R6 is H or Ci-C4 alkyl.
In yet a further embodiment, R' is H, C1-C4 alkyl or C3-C6 cycloalkyl.
In yet a further embodiment, R8 is C1-C4 alkyl.
In yet a further embodiment, R'° is imidazolyl, pyrazolyl,
triazolyl or
oxadiazolyl, each optionally substituted by 1 to 3 atoms or groups selected
from C1-C4 alkyl, C1-C4 alkoxy, cyano or halo.
In yet a further embodiment, there is provided a compound of formula
(la)
O
R3 II
N~R~
R '
Rv N
O ~ R2
O
(la)
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or a pharmaceutically acceptable salt, solvate or derivative thereof,
wherein R'; R2, R3, R4 and R5 are as defined hereinabove with respect to a
compound of formula (I), including all combinations of particular described
embodiments thereof.
In yet a further embodiment, there is provided a compound of formula
(1b)
O
4
R ~N R1
RIO N
O R2
(1b)
or a pharmaceutically acceptable salt, solvate or derivative thereof,
wherein R', R2, R4 and R5 are as defined hereinabove with respect to a
compound of formula (I), including all combinations of particular described
embodiments thereof.
It is to be understood that the invention covers all combinations of
particular embodiments of the invention as described hereinabove, consistent
with the definition of compounds of formula (I).
The compounds of the invention include compounds of formula (I) and
pharmaceutically acceptable salts, solvates or derivatives thereof (wherein
2o derivatives include complexes, polymorphs, prodrugs and isotopically-
labeled
compounds, as well as salts, solvates and salt solvates thereof), and isomers
thereof. In a further embodiment, the compounds of the invention are the
compounds of formula (I) and pharmaceutically acceptable salts and solvates
thereof, in particular the compounds of formula (I). It is to be understood
that
the aforementioned compounds of the invention include polymorphs and
isomers thereof.
Pharmaceutically acceptable salts of the compounds of formula (I)
include the acid addition and base salts thereof.
Suitable acid addition salts are formed from acids which form non-toxic
3o salts. Examples include the acetate, aspartate, benzoate, besylate,
bicarbonate, bisulphate, borate, bromide, camsylate, carbonate, chloride,
citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate,
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glucuronate, hexafluorophosphate, hibenzate, hydrobromide, hydrochloride]
hydroiodide, iodide, iseth'ionate, lactate, malate, maleate, malonate,
mesylate, ~ methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate,
orotate, oxalate, palmitate, pamoate, phosphate/hydrogen
5 phosphate/dihydrogen phosphate, saccharate, stearate, succinate, sulphate,
tartrate, tosylate and trifluoroacetate salts.
Suitable base salts are formed from bases which form non-toxic salts.
Examples include the aluminium, arginine, benzathine, calcium, choline,
diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine,
o potassium, sodium, tromethamine and zinc salts.
For a review on suitable salts, see "Handbook of Pharmaceutical Salts:
Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH,
Weinheim, Germany, 2002).
Pharmaceutically acceptable salts of compounds of formula (I) may be
prepared by one or more of three methods:
(i) by reacting the compound of formula (I) with the desired acid or base;
(ii) by removing an acid- or base-labile protecting group from a suitable
precursor of the compound of formula (I) or by ring-opening a suitable
cyclic precursor, for example, a lactone or lactam, using the desired
2o acid or base; or
(iii) by converting one salt of the compound of formula (I) to another by
reaction with an appropriate acid or base or by means of a suitable ion
exchange column.
All three reactions are typically carried out in solution. The salt may
precipitate from solution and be collected by filtration or may be recovered
by
evaporation of the solvent. The degree of ionisation in the salt may vary from
completely ionised to almost non-ionised.
The compounds of the invention may exist in both unsolvated and
solvated forms. The term 'solvate' is used herein to describe a molecular
3o complex comprising the compound of the invention and one or more
pharmaceutically acceptable solvent molecules, for example, ethanol. The
term 'hydrate' is employed when said solvent is water.
Complexes include clathrates, i.e. drug-host inclusion complexes
wherein, in contrast to the aforementioned solvates, the drug and host are
present in stoichiometric or non-stoichiometric amounts. Also included are
complexes of the pharmaceutical drug which contain two or more organic
and/or inorganic components which may be in stoichiometric or non-
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stoichiometric amounts. The resulting complexes may be ionised, partially
ionised, or non-ionised. For a review of such complexes, see J Pharm Sci, 64
(8), 1269-1288 by Haleblian (August 1975).
The compounds of the present invention may have the ability to
crystallize in more than one form, a characteristic known as polymorphism,
and all such polymorphic forms ("polymorphs") are encompassed within the
scope of the invention. Polymorphism generally can occur as a response to
changes in temperature or pressure or both, and can also result from
variations in the crystallization process. Polymorphs can be distinguished by
1 o various physical characteristics, and typically the x-ray diffraction
patterns,
solubility behavior, and melting point of the compound are used to distinguish
polymorphs.
Certain derivatives of compounds of formula (I) which may have little or
no pharmacological activity themselves can, when administered into or onto
the body, be converted into compounds of formula (I) having the desired
activity, for example, by hydrolytic cleavage. Such derivatives are referred
to
as 'prodrugs'. Further information on the use of prodrugs may be found in
'Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T
Higuchi and W Stella) and 'Bioreversible Carriers in Drug Design', Pergamon
2o Press, 1987 (ed. E B Roche, American Pharmaceutical Association).
Prodrugs in accordance with the invention can, for example, be
produced by replacing appropriate functionalities present in the compounds of
formula (I) with certain moieties known to those skilled in the art as 'pro
moieties' as described, for example, in "Design of Prodrugs" by H Bundgaard
(Elsevier, 1985).
Some examples of prodrugs in accordance with the invention include:
(i) where the compound of formula (I) contains a carboxylic acid functionality
(-CO~H), an ester thereof, for example, replacement of the hydrogen with
(Cy-C6)alkyl;
(ii) where the compound of formula (I) contains an alcohol functionality
(-OH), an ether thereof, for example, replacement of the hydrogen with
(C1-C6)alkanoyloxymethyl; and
(iii) where the compound of formula (I) contains a primary or secondary
amino functionality (-NH2 or -NHR where R ~ H), an amide thereof, for
ss example, replacement of one or both hydrogens with (Ci-Cio)alkanoyl.
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Further examples of replacement groups in accordance with the -
foregoing examples and examples of other prodrug types in accordance with
the invention may be found in the aforementioned references.
Moreover, certain compounds of formula (I) may themselves act as
s prodrugs of other compounds of formula (I).
Also included within the scope of the invention are metabolites of
compounds of formula (I), that is, compounds formed in vivo upon
administration of the drug. Some examples of metabolites in accordance with
the invention include:
(i) where the compound of formula (I) contains a methyl group, an
hydroxymethyl derivative thereof (-CH3 -> -CH20H):
(ii) where the compound of formula (I) contains an alkoxy group, an
hydroxy derivative thereof (-OR -> -OH);
(iii) where the compound of formula (I) contains a tertiary amino group, a
secondary amino derivative thereof (-NR'R2 -> -NHR' or -NHR2);
(iv) where the compound of formula (I) contains a secondary amino group,
a primary derivative thereof (-NHR' -> -NH2);
(v) where the compound of formula (I) contains a phenyl moiety, a phenol
derivative thereof (-Ph -> -PhOH); and
(vi) where the compound of formula (I) contains an amide group, a
carboxylic acid derivative thereof (-CONH2 -> COOH).
In view of the definition of R4, compounds of formula (I) contain one or
more asymmetric carbon atoms and therefore exist as two or more optical
isomers. Where a compound of formula (I) contains an alkenyl or alkenylene
group, geometric cisltrans (or Z/E) isomers are possible, and where the
compound contains, for example, a keto or oxime group or an aromatic
moiety, tautomeric isomerism ('tautomerism') may occur. It follows that a
single compound may exhibit more than one type of isomerism.
Included within the scope of the present invention are all optical
3o isomers, geometric isomers and tautomeric forms of the compounds of
formula (I), including compounds exhibiting more than one type of isomerism,
and mixtures of one or more thereof. Also included are acid addition or base
salts wherein the counterion is optically active, for example, D-lactate or L
lysine, or racemic, for example, DL-tartrate or DL-arginine.
Cisltrans isomers may be separated by conventional techniques well
known to those skilled in the art, for example, chromatography and fractional
crystallisation.
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Conventional techniques for the preparation/isolation of individual
enantiomers include chiral synthesis from a suitable optically pure precursor
or resolution of the racemate (or the racemate of a salt or derivative) using,
for example, chiral high pressure liquid chromatography (HPLC).
Alternatively, the racemate (or a racemic precursor) may be reacted
with a suitable optically active compound, for example, an alcohol, or, in the
case where the compound of formula (I) contains an acidic or basic moiety,
an acid or base such as tartaric acid or 1-phenylethylamine. The resulting
diastereomeric mixture may be separated by chromatography and/or
o fractional crystallization and one or both of the diastereoisomers converted
to
the corresponding pure enantiomer(s) by means well known to a skilled
person.
Chiral compounds of the invention (and chiral precursors thereof) may
be obtained in enantiomerically-enriched form using chromatography, typically
~5 HPLC, on an asymmetric resin with a mobile phase consisting of a
hydrocarbon, typically heptane or hexane, containing from 0 to 50%
isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine,
typically 0.1 % diethylamine. Concentration of the eluate affords the enriched
mixture.
2o Stereoisomeric conglomerates may be separated by conventional
techniques known to those skilled in the art - see, for example,
"Stereochemistry of Organic Compounds" by E L Eliel (Wiley, New York,
1994).
The present invention also includes all pharmaceutically acceptable
25 isotopically-labelled compounds of formula (I) wherein one or more atoms
are
replaced by atoms having the same atomic number, but an atomic mass or
mass number different from the atomic mass or mass number usually found
in nature.
Examples of isotopes suitable for inclusion in the compounds of the
so invention include isotopes of hydrogen, such as 2H and 3H, carbon, such as
11C' 1sC and '4C, chlorine, such as 36C1, fluorine, such as 18F, iodine, such
as
1231 and '251, nitrogen, such as'3N and 15N, oxygen, such as'S0, 170 and'$O,
phosphorus, such as 32P, and sulphur, such as 35S.
Certain isotopically-labelled compounds of formula (I), for example,
35 those incorporating a radioactive isotope, are useful in drug and/or
substrate
tissue distribution studies. The radioactive isotopes tritium, i.e. 3H, and
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carbon-14, i.e.'4C, are particularly useful for this purpose in view of their
ease-
of incorporation and ready means of detection.
Substitution with heavier isotopes such as deuterium, i.e. 2H, may
afford certain therapeutic advantages resulting trom greater metaoonc
stability, for example, increased in vivo half-life or reduced dosage
requirements, and hence may be preferred in some circumstances.
Substitution with positron emitting isotopes, such as "C, '$F,'50 and
'3N, can be useful in Positron Emission Topography (PET) studies for
examining substrate receptor occupancy.
o Isotopically-labeled compounds of formula (I) can generally be
prepared by conventional techniques known to those skilled in the art or by
processes analogous to those described in the accompanying Examples and
Preparations using an appropriate isotopically-labeled reagents in place of
the
non-labeled reagent previously employed.
~5 Pharmaceutically acceptable solvates in accordance with the invention
include those wherein the solvent of crystallization may be isotopically
substituted, e.g. D20, d6-acetone, d6-DMSO.
Preferred compounds of formula (I) include the compounds of
Examples 1-94; and pharmaceutically acceptable salts, solvates or
2o derivatives thereof. Particularly preferred compounds of formula (I)
include:
(2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl)]-2-(3-methyl-1 H-indazol-4-
yloxy)-propan-1-one;
(2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-2-[2-(2H-pyrazol-3-ylamino)-
quinolin-5-yloxy]-propan-1-one;
25 5-{(1 S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-
isoquinoline-1-carboxylic acid methylamide;
(2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-2-(5-chloro-2-methylamino-
quinolin-5-yloxy)-propan-1-one;
(2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-2-[1-(2H-pyrazol-3-ylamino)-
3o isoquinolin-5-yloxy]-propan-1-one;
4-{(1 S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-3-
methoxy-N-methyl-benzamide;
5-{(1 S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid methylamide;
35 5-{(1S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid amide;
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5-{(1S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-4-'
methoxy-pyridine-2-carboxylic acid ethylamide;
5-{(1 S)-2-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid cyclopropylamide;
5 and pharmaceutically acceptable salts, solvates or derivatives thereof.
In the general processes, and schemes, that follow: R' to R9 are as
previously defined unless otherwise stated; X is halo or hydroxy; Y is a
leaving group, such as chloro, bromo, tosylate, mesylate or hydroxy; DMF is
1o N,N-dimethylformamide; DMSO is dimethylsulphoxide; THF is
tetrahydrofuran; WSCDI is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride; DCC is N,N'-dicyclohexylcarbodiimide; HOAT is 1-hydroxy-7-
azabenzotriazole; HOBt is 1-hydroxybenzotriazole hydrate; HBTU is O-(1 H-
benzotriazol-1-yl)-N,N,N,I~P-tetramethyluronium hexafluorophosphate;
PyBOP~ is benzotriazol-1-yloxytris(pyrrolidino)phosphonium
hexafluorophosphate; PyBrOP is bromo-tris-pyrrolidino-phosphonium
hexafluorophosphate; Hunigs base is N-ethyldiisopropylamine; and
Mukaiyama's reagent is 2-chloro-1-methylpyridinium iodide.
Compounds of formula (I) may be prepared by any methods known for
2o the preparation of compounds of analogous structure.
Compounds of formula (I), and intermediates thereto, may be prepared
according to the schemes that follow.
It will be appreciated by those skilled in the art that certain of the
procedures described in the schemes for the preparation of compounds of
formula (I) or intermediates thereto may not be applicable to some of the
possible substituents.
It will be further appreciated by those skilled in the art that it may be
necessary or desirable to carry out the transformations described in the
schemes in a different order from that described, or to modify one or more of
3o the.transformations, to provide the desired compound of formula (I).
It will be still further appreciated by those skilled in the art that, as
illustrated in the schemes that follow, it may be necessary or desirable at
any
stage in the synthesis of compounds of formula (I) to protect one or more
sensitive groups in the molecule so as to prevent undesirable side reactions.
In particular, it may be necessary or desirable to protect amino groups. The
protecting groups used in the preparation of compounds of formula (I) may be
used in conventional manner. See, for example, those described in
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'Protective Groups in Organic Synthesis' by Theodora W Green and Peter G
M Wuts, third edition, (Johri Wiley and Sons, 1999), in particular chapter 7,
pages 494=653 ("Protection for the Amino Group"), incorporated herein by
reference, which also describes methods for the removal of such groups.
s The amino protecting groups boc, benzyloxycarbonyl, benzyl and
acetyl are of particular use in the preparation of compounds of formula (I)
and
intermediates thereto.
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Scheme 1
R3
~NH
HN~2 (IX) O
a
~R'
(V)
O
R3 R4 Rs
R4 ~NH s N"R'
Rw X
RIO N~2 O HN~ 2 (VII)
I O R
O (IV)
R4
X
Y
a
~ ( ) ~ (VIII)
X"R'
(V) (c) O
R3
N"R'
N ~2
R
(II)
R~ OH
(III) (f)
R3~
R4 ~N R'
RIO N
O (I)
With specific reference to scheme 1, the transformations depicted
therein may be effected as follows:
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(a)-(e) Acid-amine coupling reactions
Typically the acid chloride or acid bromide of formulae (V), (VI) or
(VIII), and the appropriate piperazine of formulae (IV), (VII) or (IX),
optionally
with an excess of an acid acceptor such as triethylamine or N ethyl-N,N
diisopropylamine, are reacted in a solvent, such as an haloalkane (e.g.
dichloromethane) or an ether (e.g. THF) at room temperature for 1-24 hours.
The reactions may conveniently be carried out by reacting the relevant
piperazine with 1.1 equivalents of the relevant acid chloride in
dichloromethane at room temperature for 1 hour.
1o In a further embodiment the acid of formulae (V), (VI) or (VIII),
activated by suitable reagents such as WSCDI/DCC and HOBdHOAt, the
appropriate piperazine of formulae (IV), (VII) or (IX), and an excess of an
acid
acceptor such as triethylamine or N ethyl-N,N diisopropylamine, are reacted
in a solvent such as an haloalkane (e.g. dichloromethane), an ether (e.g.
THF) or DMF at room temperature for 4-48 hours. The reactions may
conveniently be carried out by reacting the relevant piperazine, 1.4
equivalents of WSCDI, 1.4 equivalents of HOBt, 2.2 equivalents of
triethylamine and 1.1 equivalents of the relevant carboxylic acid in
dichloromethane at room temperature for 18 hours.
2o In yet a further embodiment, the acid of formulae (V), (VI) or (VIII), the
appropriate piperazine of formulae (IV), (VII) or (IX), and either HBTU,
PyBOP, PyBrOP or Mukaiyama's reagent, and an excess of an acid acceptor
such as triethylamine or N ethyl-N,N diisopropylamine, may be reacted in a
solvent such as an haloalkane (e.g. dichloromethane) or an ether (e.g. THF)
at room temperature for 4-24 hours. The reactions may conveniently be
carried out by reacting the relevant piperazine, 1.0 equivalents of the
relevant
carboxylic acid and 1.5 equivalents of HBTU in either dichloromethane or
DMF at room temperature for 14 hours.
(f) Nucleophilic substitution
3o i) When Y= CI, Br, mesylate, tosylate:
Typically compounds of formulae (II) and (III), with an acid acceptor,
such as triethylamine , N ethyl-N,N diisopropylamine or an alkali metal
carbonate, are reacted in a solvent such as an haloalkane (e.g.
dichloromethane), an ether (e.g. THF) or acetone at a temperature between
room temperature and reflux for 1-24 hours. The reactions may conveniently
be carried out by reacting compounds of formulae (II) and (III) with 1.0
equivalents of caesium carbonate in acetone at reflux for 14 hours.
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ii) When Y= OH:
Typically compounds of formulae (II) and (III), triphenylphosphine or
tri-o-tolylphosphine, and either diethyl azodicarboxylate or di-isopropyl
azodicarboxylate, are reacted in a solvent such as an haloalkane (e.g.
dichloromethane) or an ether (e.g. THF) at a temperature between room
temperature and reflux for 1-24 hours. The reactions may conveniently be
carried out by reacting compounds of formulae (II) and (III) with 1.2
equivalents of triphenylphosphine and 1.1 equivalents of di-isopropyl
azodicarboxylate in THF at room temperature for 14 hours.
1o Compounds of formula (VI) in reaction steps (a) and (d) may be
prepared according to scheme 1 a that follows.
Scheme 1 a
R4 R5 OH (III)
Ra Ra
OR ~ RIO OR ~ RIO OH
Y
O (J) (h) O
O
(X) (XI) (VI)
wherein R is a lower alkyl such as C1-C6 alkyl.
(g) Nucleophilic substitution
Nucleophilic substitution may be effected according to the conditions
described for step (f) hereinabove.
(h) Ester hydrolysis
Typically, compounds of formula (XI), aqueous alkali metal hydroxide
solution or aqueous hydrochloric acid solution, and an optional co-solvent
such as ethanol or dioxane, are heated at a temperature between 60 and
100°C for 1-18 hours. The reactions may conveniently be carried out by
heating compounds of formula (XI) in aqueous 1 N sodium hydroxide solution
and dioxane heated at 60 °C for 2 hours.
so Certain intermediates described above are novel compounds, and it is
to be understood that all novel intermediates herein form further aspects of
the present invention. Compounds of formulae (II), (IV) and (VII) are key
intermediates and represent a particular aspect of the present invention.
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Compounds of formulae (III), (V), (VIII), (IX) and (X) are either known
compounds or may be prepared by conventional chemistry.
According to another aspect, the invention provides the following
processes for preparing compounds of formula (I).
5 According to a first process (A), compounds of formula (I) may be
prepared by coupling an acid of formula (V)
O
X" R'
(V)
1o with a piperazine of formula (IV), under conventional acid-amine coupling
conditions.
R3
R4 ~NH
R ~O N
R2
(IV)
Conveniently, acid-amine coupling is effected under the conditions described
15 hereinabove in connection with scheme 1, steps (a)-(e).
Acoording to a second process (B), compounds of formula (I) may be
prepared by coupling an acid of formula (VI)
R4
RIO X
~ (VI)
with a piperazine of formula (VII), under conventional acid-amine coupling
conditions.
O
R3
N- _R1
HN
R2 (VII)
Conveniently, acid amine coupling is effected under the conditions
described hereinabove in connection with scheme 1, steps (a)-(e).
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16
According to a third process (C), compounds of formula (I) may be-
prepared by nucleophilic substitution of a compound of formula (II)
O
R3 ~~
a N~R~
R '
N
Y ~ R2
O (II)
with an alcohol R50H of formula (III) under conventional conditions.
Conveniently, nucleophilic substitution is effected under the conditions
described hereinabove in connection with scheme 1, step (f).
According to a fourth process (D), compounds of formula (I) may be
1o prepared from other compounds of formula (I) by functional group
interconversion under conventional conditions. For example, compounds of
formula (I) which contain an ester group may be converted to corresponding
compounds of formula (I) which contain a primary or secondary amide group
by reacting the former with ammonia or a primary amine respectively.
The compounds of the invention inhibit the interaction of gp120 with
CD4 and are therefore of use in the treatment of HIV, a retroviral infection
genetically related to HIV, and AIDS.
Accordingly, in another aspect the invention provides a compound of
formula (I) or a pharmaceutically acceptable salt, solvate or derivative
thereof
2o for use as a medicament.
In another aspect the invention provides a compound of formula (I) or a
pharmaceutically acceptable salt, solvate or derivative thereof for use in the
treatment of a HIV, a retroviral infection genetically related to HIV, or
AIDS.
In another aspect the invention provides the use of a compound of
formula (I) or of a pharmaceutically acceptable salt, solvate or derivative
thereof for the manufacture of a medicament for the treatment of a HIV, a
retroviral infection genetically related to HIV, or AIDS.
In another aspect the invention provides a method of treatment of a
mammal suffering from HIV, a retroviral infection genetically related to HIV,
or
3o AIDS which comprises treating said mammal with an effective amount of a
compound of formula (I) or a pharmaceutically acceptable salt, solvate or
derivative thereof.
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17
The compounds of the invention may be administered as crystalline or
amorphous products. They may be obtained, for example, as solid plugs,
powders, or films by methods such as precipitation, crystallization, freeze
drying, spray drying, or evaporative drying. Microwave or radio frequency
drying may be used for this purpose.
They may be administered alone or in combination with one or more
other compounds of the invention or in combination with one or more other
drugs (or in any combination thereof). Generally, they will be administered as
a formulation in association with one or more pharmaceutically acceptable
1o excipients. The term "excipient" is used herein to describe any ingredient
other than the compounds) of the invention. The choice of excipient will to a
large extent depend on factors such as the particular mode of administration,
the effect of the excipient on solubility and stability, and the nature of the
dosage form.
Pharmaceutical compositions suitable for the delivery of compounds of
the invention and methods for their preparation will be readily apparent to
those skilled in the art. Such compositions and methods for their preparation
may be found, for example, in 'Remington's Pharmaceutical Sciences', 19th
Edition (Mack Publishing Company, 1995).
2o The compounds of the invention may be administered orally. Oral
administration may involve swallowing, so that the compound enters the
gastrointestinal tract, or buccal or sublingual administration may be employed
by which the compound enters the blood stream directly from the mouth.
Formulations suitable for oral administration include solid formulations
such as tablets, capsules containing particulates, liquids, or powders,
lozenges (including liquid-filled), chews, multi- and nano-particulates, gels,
solid solution, liposome, films (including muco-adhesive), ovules, sprays and
liquid formulations.
Liquid formulations include suspensions, solutions, syrups and elixirs.
3o Such formulations may be employed as fillers in soft or hard capsules and
typically comprise a carrier, for example, water, ethanol, polyethylene
glycol,
propylene glycol, methylcellulose, or a suitable oil, and one or more
emulsifying agents andlor suspending agents. Liquid formulations may also
be prepared by the reconstitution of a solid, for example, from a sachet.
s5 The compounds of the invention may also be used in fast-dissolving,
fast-disintegrating dosage forms such as those described in Expert Opinion in
Therapeutic Patents, 11 (6), 981-986 by Liang and Chen (2001 ).
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'18
For tablet dosage forms, depending on dose, the drug may make up -
from 1 wt% to 80 wt% of the dosage form, more typically from 5 wt% to 60
wt% of the-dosage form. In addition to the drug, tablets generally contain a
disintegrant. Examples of disintegrants include sodium starch glycolate,
s sodium carboxymethyl cellulose, calcium carboxymethyl cellulose,
croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methyl cellulose,
microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose,
starch, pregelatinised starch and sodium alginate. Generally, the disintegrant
will comprise from 1 wt% to 25 wt%, preferably from 5 wt% to 20 wt% of the
1o dosageform.
Binders are generally used to impart cohesive qualities to a tablet
formulation. Suitable binders include microcrystalline cellulose, gelatin,
sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone,
pregelatinised starch, hydroxypropyl cellulose and hydroxypropyl
methylcellulose. Tablets may also contain diluents, such as lactose
(monohydrate, spray-dried monohydrate, anhydrous and the like), mannitol,
xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and
dibasic calcium phosphate dihydrate.
Tablets may also optionally comprise surface active agents, such as
2o sodium lauryl sulfate and polysorbate 80, and glidants such as silicon
dioxide
and talc. When present, surface active agents may comprise from 0.2 wt% to
5 wt% of the tablet, and glidants may comprise from 0.2 wt% to 1 wt% of the
tablet.
Tablets also generally contain lubricants such as magnesium stearate,
2s calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of
magnesium stearate with sodium lauryl sulphate. Lubricants generally
comprise from 0.25 wt°/~ to 10 wt°/~, preferably from 0.5 wt% to
3 wt% of the
tablet.
Other possible ingredients include anti-oxidants, colourants, flavours,
so preservatives and taste-masking agents.
Exemplary tablets contain up to about 80% drug, from about 10 wt% to
about 90 wt% binder, from about 0 wt% to about 85 wt% diluent, from about 2
wt% to about 10 wt% disintegrant, and from about 0.25 wt% to about 10 wt%
lubricant.
35 Tablet blends may be compressed directly or by roller to form tablets.
Tablet blends or portions of blends may alternatively be wet-, dry-, or melt-
granulated, melt congealed, or extruded before tabletting. The final
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19
formulation may comprise one or more layers and may be coated or~
uncoated; it may even be encapsulated.
The formulation of tablets is discussed in "Pharmaceutical Dosage
Forms: Tablets, Vol. 1 ", by H. Lieberman and L. Lachman, Marcel Dekker,
N.Y., N.Y., 1980 (ISBN 0-8247-6918-X).
Solid formulations for oral administration may be formulated to be
immediate and/or modified release. Modified release formulations include
delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
Suitable modified release formulations for the purposes of the
1o invention are described in US Patent No. 6,106,864. Details of other
suitable
release technologies such as high energy dispersions and osmotic and
coated particles are to be found in Verma et al, Pharmaceutical Technology
On-line, 25(2), 1-14 (2001). The use of chewing gum to achieve controlled
release is described in WO 00/35298.
The compounds of the invention may also be administered directly into
the blood stream, into muscle, or into an internal organ. Suitable means for
parenteral administration include intravenous, intraarterial, intraperitoneal,
intrathecal, intraventricular, intraurethral, intrasternal, intracranial,
intramuscular and subcutaneous. Suitable devices for parenteral
2o administration include needle (including microneedle) injectors, needle-
free
injectors and infusion techniques.
Parenteral formulations are typically aqueous solutions which may
contain excipients such as salts, carbohydrates and buffering agents
(preferably to a pH of from 3 to 9), but, for some applications, they may be
more suitably formulated as a sterile non-aqueous solution or as a dried form
to be used in conjunction with a suitable vehicle such as sterile, pyrogen-
free
water.
The preparation of parenteral formulations under sterile conditions, for
example, by lyophilisation, may readily be accomplished using standard
3o pharmaceutical techniques well known to those skilled in the art.
The solubility of compounds of the invention used in the preparation of
parenteral solutions may be increased by the use of appropriate formulation
techniques, such as the incorporation of solubility-enhancing agents.
Formulations for parenteral administration may be formulated to be
immediate and/or modified release. Modified release formulations include
delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
Thus compounds of the invention may be formulated as a solid, semi-solid, or
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thixotropic liquid for administration as an implanted depot providing modified
release of the compound. Examples of such formulations include drug-
coated sterits and PGLA microspheres.
The compounds of the invention may also be administered topically to
5 the skin or mucosa, that is, dermally or transdermally. Typical formulations
for
this purpose include gels, hydrogels, lotions, solutions, creams, ointments,
dusting powders, dressings, foams, films, skin patches, wafers, implants,
sponges, fibres, bandages and microemulsions. Liposomes may also be
used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum,
1o white petrolatum, glycerin, polyethylene glycol and propylene glycol.
Penetration enhancers may be incorporated - see, for example, J Pharm Sci,
88 (10), 955-958 by Finnin and Morgan (October 1999).
Other means of topical administration include delivery by
electroporation, iontophoresis, phonophoresis, sonophoresis and microneedle
1s or needle-free (e.g. PowderjectT"", BiojectTM, etc.) injection.
Formulations for topical administration may be formulated to be
immediate and/or modified release. Modified release formulations include
delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
The compounds of the invention can also be administered intranasally
20 or by inhalation, typically in the form of a dry powder (either alone, as a
mixture, for example, in a dry blend with lactose, or as a mixed component
particle, for example, mixed with phospholipids, such as phosphatidylcholine)
from a dry powder inhaler or as an aerosol spray from a pressurised
container, pump, spray, atomiser (preferably an atomiser using
electrohydrodynamics to produce a fine mist), or nebuliser, with or without
the
use of a suitable propellant, such as 1,1,1,2-tetrafluoroethane or
1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may
comprise a bioadhesive agent, for example, chitosan or cyclodextrin.
The pressurised container, pump, spray, atomizer, or nebuliser
so contains a solution or suspension of the compound comprising, for example,
ethanol (optionally, aqueous ethanol) or a suitable alternative agent for
dispersing, solubilising, or extending release of the compound, the
propellants) as solvent and an optional surfactant, such as sorbitan
trioleate,
oleic acid, or an oligolactic acid.
Prior to use in a dry powder or suspension formulation, the drug
product is micronised to a size suitable for delivery by inhalation (typically
less
than 5 microns). This may be achieved by any appropriate comminuting
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21
method, such as spiral jet milling, fluid bed jet milling, supercritical fluid
-
processing to form nanoparticles, high pressure homogenisation, or spray
drying.
Capsules (made, for example, from gelatin or HPMC), blisters and
cartridges for use in an inhaler or insufflator may be formulated to contain a
powder mix of the compound of the invention, a suitable powder base such
as lactose or starch and a performance modifier such as I leucine, mannitol,
or magnesium stearate. The lactose may be anhydrous or in the form of the
monohydrate, preferably the latter. Other suitable excipients include dextran,
1o glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.
A suitable solution formulation for use in an atomiser using
electrohydrodynamics to produce a fine mist may contain from 1 pg to 20mg
of the compound of the invention per actuation and the actuation volume may
vary from 1 p1 to 1 OOpI. A typical formulation may comprise a compound of the
invention, propylene glycol, sterile water, ethanol and sodium chloride.
Alternative solvents which may be used instead of propylene glycol include
glycerol and polyethylene glycol.
Suitable flavours, such as menthol and levomenthol, or sweeteners,
such as saccharin or saccharin sodium, may be added to those formulations
of the invention intended for inhaled/intranasal administration.
Formulations for inhaled/intranasal administration may be formulated
to be immediate and/or modified release using, for example, poly(DL-lactic-
coglycolic acid (PGLA). Modified release formulations include delayed-,
sustained-,
pulsed-, controlled-, targeted and programmed release.
In the case of dry powder inhalers and aerosols, the dosage unit is
determined by means of a valve which delivers a metered amount. lJnits in
accordance with the invention are typically arranged to administer a metered
dose or "puff" containing from 1 Ng to 1 Omg of the compound of the invention.
3o The overall daily dose will typically be in the range 1 pg to 200mg which
may
be administered in a single dose or, more usually, as divided doses
throughout the day.
The compounds of the invention may be administered rectally or
vaginally, for example, in the form of a suppository, pessary, or enema.
Cocoa butter is a traditional suppository base, but various alternatives may
be
used as appropriate.
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22
Formulations for rectal/vaginal administration may be formulated to be-
immediate and/or modified release. Modified release formulations include
delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
The compounds of the invention may also be administered directly to
the eye or ear, typically in the form of drops of a micronised suspension or
solution in isotonic, pH-adjusted, sterile saline. Other formulations suitable
for
ocular and aural administration include ointments, biodegradable (e.g.
absorbable gel sponges, collagen) and non-biodegradable (e.g. silicone)
implants, wafers, lenses and particulate or vesicular systems, such as
o niosomes or liposomes. A polymer such as crossed-linked polyacrylic acid,
polyvinylalcohol, hyaluronic acid, a cellulosic polymer, for example,
hydroxypropylmethylcellulose, hydroxyethylcellulose, or methyl cellulose, or a
heteropolysaccharide polymer, for example, gelan gum, may be incorporated
together with a preservative, such as benzalkonium chloride. Such
formulations may also be delivered by iontophoresis.
Formulations for ocular/aural administration may be formulated to be
immediate and/or modified release. Modified release formulations include
delayed-, sustained-, pulsed-, controlled-, targeted, or programmed release.
The compounds of the invention may be combined with soluble
2o macromolecular entities, such as cyclodextrin and suitable derivatives
thereof
or polyethylene glycol-containing polymers, in order to improve their
solubility,
dissolution rate, taste-masking, bioavailability and/or stability for use in
any of
the aforementioned modes of administration.
Drug-cyclodextrin complexes, for example, are found to be generally
useful for most dosage forms and administration routes. Both inclusion and
non-inclusion complexes may be used. As an alternative to direct
complexation with the drug, the cyclodextrin may be used as an auxiliary
additive, i.e. as a carrier, diluent, or solubiliser. Most commonly used for
these purposes are alpha-, beta- and gamma-cyclodextrins, examples of
so which may be found in International Patent Applications Nos. WO 91/11172,
WO 94/02518 and WO 98!55148.
Inasmuch as it may desirable to administer a compound of the
invention in combination with another therapeutic agent, for example, for the
purpose of treating a particular disease or condition, it is within the scope
of
the present invention that two or more pharmaceutical compositions, at least
one of which contains a compound of the invention, may conveniently be
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23
combined in the form of a kit suitable for coadministration of the-
compositions.
Thus the kit of the invention comprises two or more separate
pharmaceutical compositions, at least one of which contains a compound of
formula (I) or a pharmaceutically acceptable salt, solvate or derivative
thereof,
and means for separately retaining said compositions, such as a container,
divided bottle, or divided foil packet. An example of such a kit is the
familiar
blister pack used for the packaging of tablets, capsules and the like.
The kit of the invention is particularly suitable for administering different
o dosage forms, for example, oral and parenteral, for administering the
separate compositions at different dosage intervals, or for titrating the
separate compositions against one another. To assist compliance, the kit
typically comprises directions for administration and may be provided with a
so-called memory aid.
For administration to human patients, having a weight of about 65 to
70kg, the total daily dose of a compound of the invention is typically in the
range 1 to 10000mg, such as 10 to 1000mg, for example 25 to 500mg,
depending, of course, on the mode of administration, the age, condition and
weight of the patient, and will in any case be at the ultimate discretion of
the
2o physician. The total daily dose may be administered in single or divided
doses.
Accordingly in another aspect the invention provides a pharmaceutical
composition including a compound of formula (I) or a pharmaceutically
acceptable salt, solvate or derivative thereof together with one or more
pharmaceutically acceptable excipients, diluents or carriers.
The compounds of formula (I) and their pharmaceutically acceptable
salts, solvates and derivatives have the advantage that they are more
selective, have a more rapid onset of action, are more potent, are better
absorbed, are more stable, are more resistant to metabolism, have a reduced
'food effect', have an improved safety profile or have other more desirable
properties (e.g. with respect to solubility or hygroscopicity) than the
compounds of the prior art.
The compounds of formula (I) and their pharmaceutically acceptable
salts, solvates and derivatives may be administered alone or as part of a
combination therapy. Thus included within the scope of the present invention
are embodiments comprising coadministration of, and compositions which
contain, in addition to a compound of the invention, one or more additional
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24
therapeutic agents. Such multiple drug regimens, often referred to as-
combination therapy, may ~ be used in the treatment and prevention of
infection by human immunodeficiency virus, HIV. The use of such
combination therapy is especially pertinent with respect to the treatment and
prevention of infection and multiplication of the human immunodeficiency
virus, HIV, and related pathogenic retroviruses within a patient in need of
treatment or one at risk of becoming such a patient. The ability of such
retroviral pathogens to evolve within a relatively short period of time into
strains resistant to any monotherapy which has been administered to said
1o patient is well known in the literature. A recommended treatment for HIV is
a
combination drug treatment called Highly Active Anti-Retroviral Therapy, or
HAART. HAART combines three or more HIV drugs. Thus, the methods of
treatment and pharmaceutical compositions of the present invention may
employ a compound of the invention in the form of monotherapy, but said
~5 methods and compositions may also be used in the form of combination
therapy in which one or more compounds of the invention are coadministered
in combination with one or more additional therapeutic agents such as those
described in detail further herein.
In a further embodiment of the invention, combinations of the present
2o invention include treatment with a compound of formula (l), or a
pharmaceutically acceptable salt, solvate or derivative thereof, and one or
more additional therapeutic agents selected from the following: HIV protease
inhibitors, including but not limited to indinavir, ritonavir, saquinavir,
nelfinavir,
lopinavir, amprenavir, atazanavir, tipranavir, AG1859 and TMC 114; non
25 nucleoside reverse transcriptase inhibitors (NNRTIs), including but not
limited
to nevirapine, delavirdine, capravirine, efavirenz, GW-8248, GW-5634 and
TMC125; nucleoside/nucleotide reverse transcriptase inhibitors, including but
not limited to zidovudine, didanosine, zalcitabine, stavudine, lamivudine,
abacavir, adefovir dipivoxil, tenofovir, emtricitabine and alovudine; CCR5
3o antagonists, including but not limited to:
N-f (1 S)-3-[3-(3-isopropyl-5-methyl-4H-1,2,4-triazol-4-yl)-exo-8-
azabicyclo[3.2.1 ]oct-8-yl]-1-phenylpropyl}-4,4-
difluorocyclohexanecarboxamide or a pharmaceutically acceptable salt,
solvate or derivative thereof,
s5 methyl 1-endo-{8-[(3S)-3-(acetylamino)-3-(3-fluorophenyl)propyl]-8-
azabicyclo[3.2.1 ]oct-3-yl}-2-methyl-4,5,6,7-tetrahydro-1 H imidazo[4,5-
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c]pyridine-5-carboxylate or a pharmaceutically acceptable salt, solvate or~
derivative thereof,
methyl - 3-endo-{8-[(3S)-3-(acetylamino)-3-(3-fluorophenyl)propyl]-8-
azabicyclo[3.2.1 ]oct-3-yl)-2-methyl-4,5,6,7-tetrahydro-3H imidazo[4,5-
5 c]pyridine-5-carboxylate or a pharmaceutically acceptable salt, solvate or
derivative thereof,
ethyl 1-endo-{8-[(3S)-3-(acetylamino)-3-(3-fluorophenyl)propyl]-8-
azabicyclo[3.2.1 ]oct-3-yl}-2-methyl-4,5,6,7-tetrahydro-1 H imidazo[4,5-
c]pyridine-5-carboxylate or a pharmaceutically acceptable salt, solvate or
~o derivative thereof,
Sch-D, ONO-4128, GW-873140, AMD-887 and CMPD-167; other agents
which inhibit the interaction of gp120 with CD4, including but not limited to
BMS806 and BMS-488043; other agents which inhibit the entry of HIV into a
target cell, including but not limited to enfuviritide, T1249, PRO 542 and
15 PRO 140; integrase inhibitors, including but not limited to L-870,810; and
RNaseH inhibitors.
There is also included within the scope the present invention,
combinations of a compound of formula (I), or a pharmaceutically acceptable
salt, solvate or derivative thereof, together with one or more additional
2o therapeutic agents independently selected from the group consisting of
proliferation inhibitors, e.g. hydroxyurea; immunomodulators, such as
granulocyte macrophage colony stimulating growth factors (e.g.
sargramostim), and various forms of interferon or interferon derivatives;
other
chemokine receptor agonists/antagonists, such as CXCR4 antagonists (e.g.
25 AMD-070); tachykinin receptor modulators (e.g. NtC1 antagonists) and
various
forms of interferon or interferon derivatives; agents which substantially
inhibit,
disrupt or decrease viral transcription or RNA replication such as inhibitors
of
tat (transcriptional trans activator) or nef (negative regulatory factor);
agents
which substantially inhibit, disrupt or decrease translation of one or more
so proteins expressed by the virus (including, but not limited to, down
regulation
of protein expression or antagonism of one or more proteins) other than
reverse transcriptase, such as Tat or Nef; agents which influence, in
particular
down regulate, CCR5 receptor expression; chemokines that induce CCR5
receptor internalisation such MIP-1 a, MIP-1 [3, RANTES and derivatives
thereof; and other agents that inhibit viral infection or improve the
condition or
outcome of HIV-infected individuals through different mechanisms.
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26
Agents which influence (in particular down regulate) CCR5 receptor-
expression include immunosupressants, such as calcineurin inhibitors
(e.g. tacroliinus and cyclosporin A); steroids; agents which interfere with
cytokine production or signalling, such as Janus Kinase (JAK) inhibitors
(e.g. JAK-3 inhibitors, including 3-{(3R,4R)-4-methyl-3-[methyl-(7H
pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile) and
pharmaceutically acceptable salts, solvates or derivatives thereof; cytokine
antibodies (e.g. antibodies that inhibit the interleukin-2 (IL-2) receptor,
including basiliximab and daclizumab); and agents which interfere with cell
1o activation or cell cycling, such as rapamycin.
There is also included within the scope the present invention,
combinations of a compound of formula (I), or a pharmaceutically acceptable
salt, solvate or derivative thereof, together with one or more additional
therapeutic agents which slow down the rate of metabolism of the compound
5 of the invention, thereby leading to increased exposure in patients.
Increasing the exposure in such a manner is known as boosting. This has the
benefit of increasing the efficacy of the compound of the invention or
reducing
the dose required to achieve the same efficacy as an unboosted dose. The
metabolism of the compounds of the invention includes oxidative processes
2o carried out by P450 (CYP450) enzymes, particularly CYP 3A4 and
conjugation by UDP glucuronosyl transferase and sulphating enzymes. Thus,
among the agents that may be used to increase the exposure of a patient to a
compound of the present invention are those that can act as inhibitors of at
least one isoform of the cytochrome P450 (CYP450) enzymes. The isoforms
25 of CYP450 that may be beneficially inhibited include, but are not limited
to,
CYP1 A2, CYP2D6, CYP2C9, CYP2C19 and CYP3A4. Suitable agents that
may be used to inhibit CYP 3A4 include, but are not limited to, ritonavir,
saquinavir or ketoconazole.
It will be appreciated by a person skilled in the art, that a combination
so drug treatment, as described herein above, may comprise two or more
compounds having the same, or different, mechanism of action. Thus, by
way of illustration only, a combination may comprise a compound of the
invention and: one or more NNRTIs; one or more NRTIs and a PI; one or
more NRTIs and a CCR5 antagonist; a PI; a PI and an NNRTI; and so on.
35 In addition to the requirement of therapeutic efficacy, which may
necessitate the use of therapeutic agents in addition to the compounds of the
invention, there may be additional rationales which compel or highly
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27
recommend the use of a combination of a compound of the invention and -
another therapeutic agent, such as in the treatment of diseases or conditions
which directly result from or indirectly accompany the basic or underlying
disease or condition. For example, it may be necessary or at least desirable
to treat Hepatitis C Virus (HCV), Hepatitis B Virus (HBV), Human
Papillomavirus (HPV), opportunistic infections (including bacterial and fungal
infections), neoplasms, and other conditions which occur as the result of the
immune-compromised state of the patient being treated. Other therapeutic
agents may be used with the compounds of the invention, e.g., in order to
1o provide immune stimulation or to treat pain and inflammation which
accompany the initial and fundamental HlV infection.
Accordingly, therapeutic agents for use in combination with the
compounds of formula (I) and their pharmaceutically acceptable salts,
solvates and derivatives also include: interferons, pegylated interferons
(e.g.
peginterferon alfa-2a and peginterferon alfa-2b), lamivudine, ribavirin, and
emtricitabine for the treatment of hepatitis; antifungals such as fluconazole,
itraconazole, and voriconazole; antibacterials such as azithromycin and
clarithromycin; interferons, daunorubicin, doxorubicin, and paclitaxel for the
treatment of AIDS related fCaposi's sarcoma; and cidofovir, fomivirsen,
2o foscarnet, ganciclovir and valcyte for the treatment of cytomegalovirus
(CMV)
retinitis.
Further combinations for use according to the invention include
combination of a compound of formula (I), or a pharmaceutically acceptable
salt, solvate or derivative thereof with a CCR1 antagonist, such as BX-471; a
beta adrenoceptor agonist, such as salmeterol; a corticosteroid agonist, such
fluticasone propionate; a LTD4 antagonist, such as montelukast; a muscarinic
antagonist, such as tiotropium bromide; a PDE4 inhibitor, such as cilomilast
or roflumilast; a CO?C-2 inhibitor, such as celecoxib, valdecoxib or
rofecoxib;
an alpha-2-delta ligand, such as gabapentin or pregabalin; a beta-interferon,
so such as REBIF; a TNF receptor modulator, such as a TNF-alpha inhibitor
(e.g. adalimumab), a HMG CoA reductase inhibitor, such as a statin (e.g.
atorvastatin); or an immunosuppressant, such as cyclosporin or a macrolide
such as tacrolimus.
In the above-described combinations, the compound of formula (I) or a
pharmaceutically acceptable salt, solvate or derivative thereof and other
therapeutic agents) may be administered, in terms of dosage forms, either
separately or in conjunction with each other; and in terms of their time of
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28
administration, either simultaneously or sequentially. Thus, the
administration -
of one component agent may be prior to, concurrent with, or subsequent to
the administration of the other component agent(s).
Accordingly, in a further aspect the invention provides a
pharmaceutical composition comprising a compound of formula (I) or a
pharmaceutically acceptable salt, solvate or derivative thereof and one or
more additional therapeutic agents.
It is to be appreciated that all references herein to treatment include
curative, palliative and prophylactic treatment.
1o The invention is illustrated by the following Examples and
Preparations in which the following further abbreviations may be used:
0.88 ammonia = concentrated ammonium hydroxide solution, 0.88 SG
h = hour
min = minute
LRMS = low resolution mass spectrum
APCI+ = atmospheric pressure chemical ionisation
ESI+ = electrospray ionisation
NMR = nuclear magnetic resonance
tlc - thin layer chromatography
2o Me = methyl
Example 1
(2S~[(2R~ 4-Benzoyl-2-methyl-~perazin-1-yl)-2-(quinolin-5-yloxy)-propan-1-
one
0
Me ~N I \
Ni I O N /
\ O Me
A solution of (2S)-2-(quinolin-5-yloxy)-propionic acid sodium salt
(Preparation
2) (1.0 g, 4.6 mmol), (3R)-(3-methyl-piperazin-1-yl)-phenyl-methanone (J.
Med. Chem. (2000), 43(23), 4499) (0.94 g, 4.6 mmol), O-(1 H-benzotriazol-1-
so yl)-N,N,I~P,I~P-tetramethyluronium hexafluorophosphate (2.62 g, 6.9 mmol)
and
triethylamine (1.93 ml, 13.8 mmol) in N,N dimethylformamide (30 ml) was
stirred under a nitrogen atmosphere at room temperature for 14 hours. The
reaction mixture was diluted with dichloromethane (100 ml), washed with
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29
water (3 X 50 ml), dried (MgS04) and solvent evaporated under reduced
pressure. The crude product was purified by column chromatography on
silica gel eluting with dichloromethane: methanol (98:2) to afford the title
compound as a white solid (0.91 g).
1 H NMR (400MHz, CDC13): b: 8.90 (1 H, m), 8.60 (1 H, m), 7.70 (1 H, m), 7.55
(1 H, m), 7.20-7.40 (6H, m), 6.85 (1 H, m), 5.15 (1 H, m), 4.80-2.80 (7H, m),
1.70 (3H, d), 0.90 (3H, br s) ppm.
LRMS (APCI+) : m/z [M+H]+ 404.
1o Example 2
3-f2-l(2Rl-4-Benzovl-2-methyl-piperazin-1-vl)-1-methyl-2-oxo-ethoxv]-2-
methyl-benzoic acid methyl ester
0
Me ~N ~ \
Me0 \ O N
O Me O Me
Cesium carbonate (0.39 g, 1.2 mmol) was added portionwise to a solution of
1-[(2R)-4-benzoyl-2-methyl-piperazin-1-yl]-2-bromo-propan-1-one
(Preparation 3) (0.41 g, 1.2 mmol) and 3-hydroxy-2-methyl-benzoic acid
methyl ester (Tet. Lett. (2000), 41 (11 ), 1741 ) (0.2 g, 1.2 mmol) in acetone
(10
ml) and the mixture heated under reflux for 14 hours. The cooled reaction
2o mixture was evaporated to dryness under reduced pressure and the residue
partitioned between dichloromethane (50 ml) and water (10 ml). The organic
phase was separated and the aqueous phase extracted with dichloromethane
(3 X 30 ml), the combined organic extracts were dried (MgS04) and solvent
evaporated under reduced pressure to afford the title compound as a white
solid (0.5 g).
1 H NMR (400MHz, CDC13): S: 7.50-7.30 (6H, m), 7.15 (1 H, m), 6.95 (1 H, m),
4.95 (1 H, m), 4.80-2.80 (7H, m), 3.90 (3H, s), 2.40 (3H, s), 1.65 (3H, d),
1.20
(3H, br s) ppm.
LRMS (APCI+) : mlz [M+H]+ 425.
3o Examples 3-12 were prepared by the method described above for
Example 1, using the corresponding acid of formula (VI) and piperazine of
formula (VI I). LRMS was by APCI+ and data quoted are for [M+H]+.
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Mass
Example
Structure Spec.
Number Data
0
/
3 \ I Me ~N I \ 386
CI O /
O Me
O
Me ~N \
4 \ ~ N ~ , 380
Me O
Me O Me
O
Me ~N \
5 \ I N ~ / 408
~o
HN~ ~ Me
O F
Me ~N \
6 ~ / ~ _ N ~ / 422
\ ~ ~ ~ Me
O
Me ~N \ F
N ~ / 422
N, Y ~O
\ O Me
O
Me ~N \
8 ~ / I _ N ~ / 422
N, Y O F
\ ~ Me
O
Me ~N Nw
9 ~ / ~ N ~ / 405
N, Y ~O
\ O Me
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31
Mass
Example .
Structure Spec.
Number Data
0
Me ~N ~ ~ N
N ~ o N ~ 405
\ ~ O Me
O O
Me~H \ ~ Me ~''N ~ \' 440
11
OMe O Me
O
Me ~N ~ \
12 \ o N ~ 407
HN
N- O Me
Me
Examples 13-45 were prepared by the method described above for
Example 2, using the corresponding compound of formula (II) and R5~H.
LRMS was by APCI+ and data quoted are for [M+H]+, unless otherwise
s stated.
Mass
Example
Structure Spec.
Number Data
o
13 \ I Me ~N I j 386
Ci O
O Me
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Mass
Example
Structure Spec.
Number Data
0
/
14 \ I Me ~N I j 380
Me O
Me O Me
O
15 \ I Me ~N I j 366
Me O
Me O
O
16 \ ~ Me ~N I ~ 391
Me O
CN O Me
O
17 \ I Me ~N I ~ 383
O
OMe O Me
O
18 ~ ~ Me ~N ~ , 393
o O
=--N O Me
O
Me ~N
19 w I N ~ , 419
N
O Me
O
20 \ I Me ~N I j 367
-O
Me O Me
O
/ N Me ~N
21 ~ ~ N ~ , 368
O
Me O Me
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33
Mass
Example
Structure Spec.
Number Data
0
Me ~N \ 403
22 N~ ~ o N ~ ~ (ESI+,
\ I O Me M+)
O
F
Me ~N \
23 / I N ~ , 421
\ N O Me
Me p
380
24 \ ( Me ~N I \ (ESI+,
~ M+)
Me O Me
O
25 N~ ) / Me ~N ~ / 393
H
N O
O Me
O
26 \ ~ / Me ~N I j 402
O
O Me
O
Me ~N
27 / ~ N ~ , 403
i I~ ,o
\ N O Me
O
Me ~N
28 ~ ~ o N ~ ~ 403
NJ O Me
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34
Mass
Example .
Structure Spec.
Number Data
0
Me ~N \
29 / I N ~ / 402
~o
\ I O Me
O O
Me~H / ~ Me ~N ~ \ 424
30 Me ~ O~N~ /
O M'e
O O
Me~H / ~ Me ~N ~ \ 440
31 ~ o~N
OMe ~O( M~e
O O
Et~H / ~ Me ~N ~ ~ 454
32 ~ o~N
OMe O M~e
O
Me ~N ~ \
33 \ o N / 407
HN
N- O Me
Me
O
Me ~N ~ \
34 HN o / 407
=-N O Me
Me
O
35 \ ~ Me ~N I \ 368
Me O /
O Me
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Mass
Example
Structure Spec.
Number
Data
Me p
O
36 ~ ~ , Me ~N ~ \ 434
O
O Me
O
Me N
37 ~ ~ j Me ~N I j 417
O
IOI Me
O
Me ~N
38 ~ , N ~ , 403
O
O Me
O
39 N \ ~ ~ Me ~N I ~ 403
O
O Me
O
Me ~N
Me,N ~ O~N / 421
N- O Me
Me
O O
41 H2N \ I Me ~N ~ j 426
O
OMe O Me
O O
42 HEN \ I Me ~N ( % 410
Me O
IOI Me
O O
43 H2N \ ~ Me ~N I j 426
Me0 O
IOI Me
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36
Mass
Example
Structure Spec.
Number Data
0
Me ~N
44 ~ o N ~ ~ 396
H2N O O Me
O
N
Me N
45 ~ ~ , ~ ~ , 405
O
O Me
Example 46
(2Sl-1-f(2R)-4-Benzovl-2-methvl-piperazin-1-VI1-2-f2-methox -4- 2H-
j1 2 4]triazol-3-ylLphenoxy~-propan-1-one
O
NwN CHs ~N I \
H N /
O
O CHs
CHs
The compound of Preparation 10 (125mg, 0.26mmol) was added portionwise
to a stirred solution of hydrazine monohydrate (25pL, 0.52mmol) in glacial
acetic acid (4mL). The resulting solution was heated at 90°C for 3
hours. The
1o reaction mixture was then evaporated under reduced pressure and the
residual colourless oil was partitioned between dichloromethane (50mL) and
saturated sodium hydrogen carbonate solution (8mL). The organic phase was
separated and the aqueous phase was re-extracted with dichloromethane
(30mL). The combined organic phases were dried over magnesium sulfate
~5 and concentrated in vacuo to yield a colourless foam. The foam was dried
under reduced pressure to afford the title compound in 94% yield, 110mg.
LRMS (ES+): mlz [M+H]+ 450
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37
Example 47
1-f(2R -4-Benzoyl-2-methyl-piperazin-1-yl]-2-f2-methyl-3 ~1-methyl-1 H-
pyrazol-3 yl)-phenoxyLpropan-1-one
O
CH3 ~N
~N~ ~ O N
H3C-N
CH3 O CH3
The compounds of Preparations 17 (100mg, 0.53mmol) and 25 (180mg,
0.53mmol) and cesium carbonate (172.7mg, 0.53mmol) in acetone (8mL)
were heated under reflux for 3.5 hours. The reaction mixture was then cooled
and evaporated under reduced pressure. The residue was partitioned
1o between dichloromethane (30mL) and water (8mL). The organic phase was
separated and the aqueous phase was re-extracted with further
dichloromethane (30mL). The combined organic extracts were dried over
magnesium sulfate and concentrated in vacuo to give a light brown oil. The oil
was chromatographed using an Isolute~ Flash silica column, eluting with
dichloromethane:methanol, 99:1 to 97:3, to give a colourless oil. This oil was
azeotroped with dichloromethane to afford the title compound as colourless
foam in 89% yield, 210mg.
LRMS (ES+): m/z [M+H]+ 447
2o Example 48
1-[~(2R)-4-Benzoyl-2-methyl-piperazin-1-y~2-methyl-3-[1,2,4~oxadiazol-5-yl-
phenoxy~propan-1-one
CH3 ~N
O ~ O N
N
~IN CH3 O CH3
The compound of Preparation 29 (280mg, 0.62mmol) was heated in a mixture
of glacial acetic acid (3mL) and dioxane (3mL) at 90°C, for 3.5 hours.
The
mixture was then allowed to cool and was evaporated under reduced
pressure to give a yellow oil. The oil was dissolved in ethyl acetate (50mL)
and washed with 10% sodium carbonate solution (lOmL) and water (lOmL).
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38
The organic phase was dried over magnesium sulfate and concentrated in-
vacuo to give a colourless foam. The foam was purified on an Isolute~ flash
silica column, eluting with dichloromethane:methanol, 99:1 to 97:3, to give a
colourless oil. The oil was azeotroped with dichloromethane to afford the
title
s compound as a colourless foam in 44% yield, 118mg.
LRMS (ES-): m/z [M-H]-433
Example 49
(2S)-1~[(2R)-4-Benzoyl-2-meth iLl-piperazin-1-yll-2-[~2H-pyrazol-3-yl)-
io phenoxy]'-propan-1-one
O
CH3 ~N
N
O
O CH3
H ~
N
(3R)-(3-methyl-piperazin-1-yl)-phenyl-methanone [(114mg, 0.56mmol), J.
Med. Chem. 43(23), 4499; 2000], 3-(diethoxyphosphoryloxy)-1,2,3-
benzotriazin-4(3H)-one (167.5mg, 0.56mmol) and triethylamine (78pL,
15 0.56mmol) were added to a solution of the compound of Preparation 23
(130mg, 0.56mmol) in tetrahydrofuran (8mL) and the resulting solution was
stirred at room temperature for 48 hours. The mixture was then diluted with
ethyl acetate (30mL) and washed with water (lOmL), 10% citric acid (lOmL),
water (lOmL) and sodium hydrogen carbonate solution. The organic phase
2o was dried over magnesium sulfate and concentrated in vacuo. The residue
was purified on an Isolute~ flash silica column, eluting with
dichloromethane:methanol, 98:2 to 96:4, to give a colourless foam. The foam
was azeotroped with dichloromethane and dried under reduced pressure to
afford the title compound in 85% yield, 198mg.
25 LRMS (ES-): m/z [M-H]-417
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39
Example 50
(2S)-1-[~2R~4-Benzoyl-2-metal-~perazin-1-yl]-2-[2-methyl-3-(2H-pyrazol-3-
yl~ phenoxy]-propan-1-one
0
CH3 ~N
N /
\ O
O CH3
~CH3
H ~
N
The title compound was prepared by the method described above for
Example 49, using the compound of Preparation 24 and
(3R)-(3-methyl-piperazin-1-yl)-phenyl-methanone ( J. Med. Chem. 43(23),
4499; 2000).
LRMS (ES-): m/z [M-H]-431
Example 51
1-~(2R~4-Benzoyl-2-methyl-piperazin-1-yl]-2-[~1 H-imidazol-2-yl)-2-methoxy-
~ahenoxy]-propan-1-one
0
CH3 ~N ~ \
N /
\ 0
H I O CH3
N /
~~N CH3
The compounds of Preparation 33 (30mg, 0.16mmol) and Preparation 25
(53.5mg, 0.16mmol) and cesium carbonate (51.3mg, 0.16mmol) in N,N-
dimethylformamide (4mL) were heated at 80°C for 18 hours. The reaction
mixture was allowed to cool and was then evaporated under reduced
2o pressure and the residue was partitioned between dichloromethane (30mL)
and water (8mL). The aqueous phase was separated and was re-extracted
with dichloromethane (30mL). The combined organic extracts were dried over
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magnesium sulfate and concentrated in vacuo. The residue was purified on
an Isolute~ flash silica column, eluting with dichloromethane:methanol, 99:1
to
95:5, to afford the title compound as a beige foam in 74% yield, 52mg.
LRMS (ES+): m/z [M+H]+ 449
5
Examples 52 to 53
The following compounds, of the general formula shown below, were
prepared by the method described above for Example 51, using the
o compound of Preparation 25 and the appropriate phenol.
0
CH3 ~N
N
O
0 CH3
R O
CH3
No. R~ Data
52 ~ N LRMS (ES-): m/z [M-H]~
447
N\
53 j LRMS (ES-): mlz [M+Na]+
~N 471
~
~5 Examples 54 to 57
The following compounds, of the general formula shown below, were
prepared by the method described above for Example 51, using the
compound of Preparation 25 and the appropriate phenol.
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41
0
CH3 ~N
R\O N /
O CH3
No. R5 LRMS (APCI+~: m/z
[M+H]+
54 j H3 429
s \
0
H3C~
55 HN 452
o'
0
H3C~
56 I H3 433
57 ~ / ~ 482
H2N~ //
/% /
0 ~ _
Example 54: phenol prepared as described in Preparation 67
Example 55: phenol prepared as described in Preparation 68
Example 56: 6-methoxy-2-naphthol is commercially available
Example 57: 5-hydroxy-1-naphthalene sulfonamide is commercially available
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42
Example 58
1-f(2Rl-4-Benzovl-2-methyl-aioerazin-1-vll-2-(4-methanesulfonvl-2-methox
i~henoxy~-propan-1-one
O O
_1I
H3C II / CH3 ~N \
o \ ~ N
-o
H C~~ O CH3
3
A solution of oxone (215mg, 0.35mmol) in water (2mL) was added to a
solution of the compound of Example 54 (100mg, 0.23mmol) in methanol
(4mL) and the mixture was stirred for 18 hours at room temperature. The
solvent was then evaporated under reduced pressure and the residue was
partitioned between water (20mL) and dichloromethane (20mL). The aqueous
layer was separated and re-extracted with dichloromethane (30mL). The
combined organic extracts were dried over magnesium sulfate and
concentrated in vacuo. The residue was purified on an Isolute~ flash silica
column, eluting with dichloromethane:methanol, 95:5 to afford the title
com~aound as a white solid in 78% yield, 80mg.
LRMS (APCI+): m/z [M+H]+ 481
Example 59
N-~4-[2~2R)-(4-Benzoyl-2-methyl-piperazin-1-yl)-1-methyl-2-oxo-ethoxy]-3-
methox~phenyl}-methanesulfonamide
O H I Hs O
H3C ~ N / O CH3 ~N \
\ N /
O
O CH3
Triethylamine (0.08mL, 0.57mmol) was added to a solution of the compound
of Preparation 30 (150mg, 0.38mmol) in dichloromethane (3mL) and the
mixture was cooled in an ice bath. Methanesufonyl chloride (0.03mL,
0.42mmol) was added dropwise and the reaction mixture was allowed to stir
at room temperature for 18 hours. The mixture was then diluted with further
dichloromethane (20mL) and washed with water. The organic phase was
dried over magnesium sulfate and concentrated in vacuo. The residue was
then purified by column chromatography on silica gel, eluting with ethyl
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43
acetate:methanol, 98:2, to afford the title compound as a white solid in 49%
yield, 88mg.
LRMS (APCI+): m/z [M+H]+ 476
Example 60
,(2S -1~-f(2R -4-Benzoyl-2-methpiperazin-1-y~-2-(2-chloro-quinolin-5-yloxy~-
propan-1-one
0
CH3 ~N
N/ I O N /
CI \ O CH3
o A solution of the compound of Preparation 46 (1.8g, 4.3mmol) in
phosphorous oxychloride (5mL) was heated at 100°C for two hours. The
reaction mixture was then cooled to room temperature and evaporated under
reduced pressure. The residue was dissolved in dichloromethane (20mL)
and washed with saturated aqueous sodium hydrogen carbonate solution
(1 OmL) and water (2x1 OmL). The aqueous phase was basified with 1 M
sodium hydroxide solution and was re-extracted with ethyl acetate. The
combined organic extracts were then dried over magnesium sulfate and
concentrated in vacuo. The residue was purified by column chromatography
on silica gel, eluting with pentane:ethyl acetate 20:80, 0:100 to afford the
title
2o compound as a pale yellow solid in 31 % yield, 580mg.
'H NMR (400MHz, CDCI3) b: 1.00-1.40(m, 3H), 1.70-1.80(m, 3H), 2.80-4.80
(m, 7H), 5.20(m, 1 H), 6.90(m, 1 H), 7.20-7.45(m, 6H), 7.55-7.70 (m, 2H),
8.55(d, 1 H)
LRMS (APCI+): m/z [M+H]+ 438
Examples 61 to 62
A mixture of the compound of Example 60 (30mg, 0.069mmol), tetrabutyl
ammonium fluoride (18mg, 0.14mmol), triethylamine (96pL, 0.69mmol) and
3o the appropriate amine (0.69mmol) in dimethylsulfoxide (1 mL) was heated at
100°C for 24 hours. The reaction mixtures were then purified directly
by HPLC
using a Phenomenex Luna C18 system, eluting with
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44
water/acetonitrile/trifluoroacetic acid (5:95:0.1):acetonitrile, 95:5 to 5:95,
to'
afford the desired compounds of general formula shown below:
CH3
r
\ o
O CH3
( \
N /
t R6,NH
No. R6 LRMS (APCI+): m/z [M-H]+
61 461
H3C'
62 ~H3 461
H3C
Example 63
1-f (2R)-4-Benzoyl-2-methyl-piperazin-1-yl)-2-(1-chloro-isoguinolin-5-ylox~
propan-1-one
CH3
O
\ ~ :5
N / /
CI
1o The title compound was prepared by the method described above for
Example 60, using the compound of Preparation 63 and phosphorus
oxychloride.
LRMS (APCI+): m/z [M+H]+ 438
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Examples 64 to 65
A mixture of the compound of Example 63 (45mg, 0.1 mmol), tetrabutyl
5 ammonium fluoride (27mg, 0.2mmol), triethylamine (140pL, 1mmol) and the
appropriate amine (1 mmol) in dimethylsulfoxide (1 mL) was heated at
130°C
for 24 hours. The reaction mixtures were then purified directly by HPLC using
a Phenomenex Luna C18 system, eluting with water/acetonitrile/trifluoroacetic
acid (5:95:0.1 ):acetonitrile, 95:5 to 5:95 to afford the desired compounds of
1o general formula shown below:
0
CH3 ~N
N
O
O CH3
R~
N N
R'
No. R6 R' LRMS (APCI+): m/z [M-HJ+
64 N~N H 485
65 CH3 CHI 447
20
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46
Example 66
(2Sl-2-(2-Amino-auinolin-5-vloxv)-1-f (2R)-4-benzovl-2-methyl-piperazin-1-
propan-1-one
p
CH3 ~N ~ \
N /
\ O
O CH3
N /
NH2
s 10% Pd/C (cat) and ammonium formate (l0mg, 0.15mmol) were added to a
solution of the compound of Preparation 49 (l5mg, 0.03mmol) in ethanol
(1 mL) and the mixture was heated at 70°C for 2 hours. Additional
ammonium
formate (l0mg, 0.15mmol) and Pd(OH)2 (cat) were added and the reaction
mixture was heated under reflux for a further 18 hours. The mixture was then
1o cooled and filtered through Arbocel~, washing through with ethanol (lOmL).
The filtrate was concentrated in vacuo and the residue was purified by HPLC
using a Phenomenex Luna C18 system, eluting with
water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile, 95:5 to
5:95, to
afford the title compound in 77% yield, 9.7mg.
15 LRMS (ES+): m/z [M+H]+ 419
Example 67
l2Sl-1-f (2R)-4-Benzovl-2-methyl-piperazin-1-vll-2-(8-chloro-auinolin-5-vlox
propan-1-one
0
/ ( CH3 ~N
N/ o N /
O CH3
A mixture of the compound of Preparation 42 (30mg, 0.07mmol) and N-
chlorosuccinimide (11 mg, 0.08mmol) in acetonitrile (3mL) was heated at
40°C
for 48 hours. The solvent was then evaporated under reduced pressure and
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47
the residue was dissolved in ethyl acetate (5mL) and washed with water
(2x5mL). The organic phase~was dried over magnesium sulfate and
concentrated in vacuo to give a yellow residue. The residue was purified by
HPLC using a Phenomenex Luna C18 system, eluting with
water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile, 95:5 to
5:95, to
afford the title compound as a solid in 32% yield, 9.7mg.
LRMS (APCI+): m/z [M+HJ+ 438
Examples 68 to 69
The following compounds, of the general formula shown below, were
prepared by the method described above for Example 67, using the
appropriate starting material and N-chlorosuccinimide.
o
C~ /
Cf-13 ~N
N/ O N /
\ O CH3
No. X LRMS (APCI+): m/z [M-HJ+
68 N 439
69 C-F 456
Examples 68 and 69 were purified by column chromatography on silica gel,
eluting with dichloromethane:methanol, 99:1 to 96:4
Examples 70 and 71
The compounds were prepared from the compound of Preparation 60 and 1-
chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate),
using a similar preparation to Example 67. The compounds were purified by
HPLC using a Phenomenex Luna C18(2) column 150x15mm (10 micron
particle size, 100A porosity), using a 2 solvent eluent of
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acetonitrile:waterarifiuoroacetic acid (5:95:0.1 ) [solvent A] and acetonitile
-
[solvent B]. A solvent gradient is run at a flow-rate of 20 ml/min as in the
table
below. -
Time min) %B
0 5
0.6 5
9.5 95
10.5 95
10.6 5
12 5
No Structure HPLC LRMS (APCI+):
retention time m/z [M+H]+
min.
70 4.99 422
0
F
I CH3 ~N I \
N/ I O N /
\ O CHI
71 5.77 422
0
/ F
I CH3 ~N I \
N/ I O N /
\ O CH3
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Example 72
~2S)-1-f (2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-2-(8-methyl-quinolin-5-yloxy~-
propan-1-one
0
H3C
I CH3 ~N \
N~ O N I
\ O CH3
A mixture of the compound of Preparation 44 (200mg, 0.4mmol), methyl
boronic acid (74mg, l.2mmol), bis(triphenylphosphine)palladium(II) chloride
(29mg, 0.04mmol) and potassium carbonate (229mg, l.6mmol) in N,N-
dimethylformamide (4mL) was heated at 100°C for 18 hours. Tlc analysis
1o showed that the reaction was incomplete and so additional amounts of methyl
boronic acid (74mg) and bis(triphenylphosphine)palladium(II) chloride (29mg)
were added, and the mixture was allowed to stir for a further 24 hours at
100°C. The reaction mixture was then filtered through Arbocel~, washing
through with ethyl acetate (25mL) and water (3x20mL). The layers were
separated and the aqueous layer was re-extracted with ethyl acetate. The
combined organic solutions were dried over magnesium sulfate and
concentrated in vacuo to afford a yellow residue. The residue was purified by
HPLC using a Phenomenex Luna C18 system, eluting with
water/acetonitrile/trifluoroacetic acid (5:95:0.1):acetonitrile, 95:5 to 5:95.
The
2o appropriate fraction was then azeotroped with diethyl ether to afford the
title
compound as a yellow solid in 19% yield, 33mg.
LRMS (APCI+): m/z [M+H]+ 418
30
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Example 73
(2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-,Lrll-2-[8-~(2H-pyrazol-3-yl)-
quinolin-5-yloxy~-propan-1-one
5 4M Hydrochloric acid (2mL) was added to a suspension of the compound of
Preparation 45 (43mg, 0.07mmol) in methanol (2mL), and the resulting
solution was stirred at room temperature for 18 hours. The reaction mixture
was then evaporated under reduced pressure and the residue was dissolved
in water and washed twice with dichloromethane. The combined organic
10 layers were dried over magnesium sulfate and concentrated in vacuo to give
a yellow residue. The residue was purified by column chromatography on
silica gel, eluting with dichloromethane:methanol, 98:2, to afford the title
compound as a yellow solid in 68% yield, 23mg
LRMS (APCI+): m/z [M+H]+ 470
Example 74
(~S)-1-f (2R)-4-Benzovl-2-methyl-piperazin-1-vll-2-f2-(2H-avrazol-3-vlaminol-
quinolin-5-yloxy]-propan-1-one
0
CH3 ~N
~~N N~ ~ O N
\ O CH3
N
H
A mixture of the compound of Example 60 (50mg, 0.11 mmol), 3-
aminopyrazole (29mg, 0.33mmol) and triethylamine (48pL, 0.33mmol) in 2-
propanol (0.5mL) was heated at 90°C in a ReactivialT"". After 18 hours,
cesium fluoride (33mg, 0.22mmol) was added and the mixture was heated at
100°C for a further 48 hours. The reaction mixture was then filtered
and
concentrated in vacuo to afford an oily residue. The residue was purified by
HPLC using a Phenomenex Luna C18 system, eluting with
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water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile, 95:5 to
5:95, to
afford the title compound as a yellow solid 3mg.
LRMS (APCI+): m/z [M+H]+ 485
Example 75
5-~(1S~2-[(2R -4.-Benzo~rl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy~
1 H~uinolin-2-one
O
/ ~ CH3 ~N ~ \
HN \ O N /
O / O CH3
1o The compound of Example 60 (200mg, 0.46mmol) and acetic acid (3mL)
were dissolved in water and the solution was heated under reflux for 24
hours. The reaction mixture was then diluted with water and washed with
dichloromethane (3x10mL). The combined organic extracts were dried over
magnesium sulfate and concentrated in vacuo to give a yellow oil. The oil was
purified by column chromatography on silica gel, eluting with
dichloromethane:methanol, 95:5 to afford the title compound as pale yellow
solid in 40% yield, 76mg.
LRMS (APCI+): m/z [M+H]+ 418
2o Example 76
~(1 S)-2-~(2R)-4.-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethox~r~8-
chloro-1 H-quinolin-2-one
O
C~
CH3 ~N ~ \
HN \ O N
O / O CH3
The title compound was prepared by the method described above for
Example 67, using the compound of Example 75 and N-chlorosuccinimide.
LRMS (APCI+): m/z [M+H]+ 454
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Example 77
(2S)-1 ~[(2F~-4-Benzoyl-2-methyl-piperazin-1-yll-2-(~-methoxy-quinolin-5-
ylox~r -propan-1-one
0
CH3 ~N I \
Ni I 0 N /
H3C~0 \ O CH3
A mixture of the compound of Example 60 (20mg, 0.04mmol) and sodium
methoxide (0.5M in methanol, 0.7mL) in N,N-dimethylformamide (0.5mL) was
heated in a ReactivialT"" at 70°C for 6 hours. The reaction mixture was
1o cooled, and evaporated under reduced pressure. The residue was dissolved
in dichloromethane (5mL) and washed with brine (5mL) and water (2x5mL).
The organic phase was dried over magnesium sulfate and concentrated in
vacuo. The residue was dissolved in dimethylsulfoxide (0.5mL) and was
purified by HPLC using a Phenomenex Luna C18 system, eluting with
water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile, 95:5 to
5:95. The
appropriate fraction was then azeotroped with diethyl ether to afford the
title
compound as a yellow solid in 40% yield, 7mg.
LRMS (APCI+): m/z [M+H]+ 434
2o Example 78
5-((1 S)-2-f(2R)-4-Benzovl-2-methvl-aioerazin-1-vll-1-methyl-2-oxo-ethox
isoquinoline-1-carboxylic acid methyl ester
O
O / ~ CH3 ~N
H3C~0 / O N /
N J O CH3
The compound of Preparation 65 (80mg, 0.18mmol), dichloro-
bis(triphenylphosphine)palladium (l5mg, 0.018mmol) and triethylamine
(51 pL, 0.37mmol) were dissolved in methanol (5mL) and transferred to a
sealed vessel. The vessel was heated to 100°C and the mixture was
stirred
under 100psi of carbon monoxide gas for 42 hours. The reaction mixture was
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53
then filtered through Arbocel~, washing through with methanol and the filtrate
was concentrated in vacuo.~ The residue was purified on an Isolute~ flash
silica colur~in, eluting with dichloromethane:methanol, 99:1 to afford the
title
compound as a white foam in 43% yield, 36mg.
LRMS (APCI+): m/z [M+H]+ 462
Example 79
5~2-~2R -4-Benzoyl-2-methyl-pi~~erazin-1-yl]-1-methyl-2-oxo-ethoxy}-
quinoline-2-carboxylic acid meth la~_ mide
0
CH3 ~N ~ \
N
0 O CH3
~NH
H3C
A mixture of the compound of Preparation 48 (20mg, 0.05mmol),
methylamine (33% in ethanol, 30mg, 0.5mmol), O-(1 H-benzotriazol-1-yl)-
N,N,N',IV-tetramethyluronium hexafluorophosphate (2mg, 0.08mmol) and
triethylamine (62pL, 0.5mmol) in dichloromethane (1 mL) was stirred at room
temperature for 18 hours. Additional methylamine (30mg, 0.5mmol) and
triethylamine (62 pL, 0.5mmol) were added to the reaction mixture and stirring
continued for a further 18 hours. The reaction mixture was then diluted with
dichloromethane and washed with water (2x5mL) and brine (5mL). The
organic phase was dried over magnesium sulfate and concentrated in vacuo
2o to give a yellow residue. The residue was purified by column chromatography
on silica gel, eluting with dichloromethane:methanol, 96:4, to afford the
title
compound as a white solid in 13% yield, 3mg.
LRMS (APCI+): m/z [M+H]+ 461
30
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54
Example 80
5-f(1 Sl-2-f(2R)-4-Benzovl-2-r~nethvl-aiperazin-1-vll-1-methyl-2-oxo-ethox
isoquinoline-1-carbox~rlic acid methylamide
O
O / ~ CHs ~N
HsCwN / ~ N /
H
N J O CH3
The title compound was prepared by the method described above for
Example 79, using the compound of Example 78 and methylamine.
LRMS (APCI+): m/z [M+H]+ 461
Example 81
1o 4-~~~1 S)-2-[(2R -4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy)-
N-
methyl-3-trifluoromethoxy-benzamide
H
H3C\N CH3
N
O
FI
F
The compound of Preparation 59 (100mg, 0.2mmol) and methylamine (33%
in ethanol, 4mL) were heated at 90°C in a ReactivialT"" for 18 hours.
The
solvent was then evaporated under reduced pressure and the residue was
purified by column chromatography on silica gel, eluting with
dichloromethane:methanol, 99:1 to 97:3, to afford the title compound as a
pale orange foam in quantitative yield, 100mg.
LRMS (APCI+): m/z [M+H]+ 494
25
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Example 82
~2S)-1-f(2R -4-Benzoyl-2-methyl-piperazin-1-~r11-2-(8-chloro-2-methylamino-
quinolin-5~iloxy~propan-1-one
/ ~ CHs
N ~ I ~o
H3CwN \ 0
H
5 A mixture of the compound of Preparation 52 (60mg, 0.12mmol),
methylamine hydrochloride (86mg, l.2mmol), cesium fluoride (39mg,
0.24mmol) and triethylamine (0.18mL, 1.2mmol) in dimethylsulfoxide (1 mL)
was heated in a microwave for 8 minutes at 170°C. The reaction mixture
was
then filtered and the filtrate was purified by HPLC using a Phenomenex Luna
1o C18 system, eluting with water/acetonitrile/trifluoroacetic acid
(5:95:0.1 ):acetonitrile, 95:5 to 5:95. The appropriate fraction was then
azeotroped with diethyl ether to afford the title compound as a brown solid in
19% yield, 10.8mg.
LRMS (APCI+): m/z [M+H]+ 467
Example 83
1-f (2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-~1-methylamino-isoquinolin-5~i
loxy~propan-1-one
0
CH3 ~N ( \
NI /
O
\ \ 0 CH3
//~
,NH
H3C
2o The title compound was prepared by the method described above for
Example 82, using the compound of Example 63 and methylamine.
LRMS (APCI+): m/z [M+H]+ 433
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Example 84
(2S~1-f 2R)-4-Benzoyl-2-methyl-~perazin-1-yl]-2-(8-chloro-2-dimethylamino-
auinolin-5~ilo~)-pr~an-1-one
_ / ~ CH3 .
N / ( w0
H3C~ \ O CH3
N
CH3
A mixture of the compound of Preparation 52 (50mg, 0.1 mmol),
dimethylamine (86mg, 1 mmol), cesium fluoride (32mg, 0.2mmol) and
triethylamine (0.15mL, 1 mmol) in dimethylsulfoxide (4mL) was heated at
170°C in a ReactivialT"" for 18 hours. The reaction mixture was then
filtered
1o and the filtrate was purified by HPLC using a Phenomenex Luna C18 system,
eluting with water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile,
95:5 to
5:95, to afford the title compound as a white solid, 5mg.
LRMS (APCI+): m/z [M+H]+ 481
Example 85
(2S)-1-f(2R)-4-Benzoyl-2-methyl-~perazin-1-yll-2-f1- 2H-pyrazol-3-ylamino)-
isoquinolin-5-Lrloxy]-propan-1-one
O
/ ( CH3 ~N
H
N / O N /
W
N J O CH3
N
2o A mixture of the compound of Example 63 (45mg, 0.1 mmol), tetrabutyl
ammonium fluoride (27mg, 0.2mmol), triethylamine (0.14mL, 1 mmol) and 3-
aminopyrazole (83mg, 1 mmol) in dimethylsulfoxide (1 mL) was heated at
100°C for 24 hours. The reaction mixture was then purified directly by
HPLC
using a Phenomenex Luna C18 system, eluting with water/acetonitrile/formic
acid (5:95:0.1 ):acetonitrile, 95:5 to 5:95, to afford the title compound in
39%
yield, l9mg.
LRMS (APCI+): m/z [M+H]+ 485
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Example 86
~2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1- rLl)]-2-(3,6-dimethyl-1 H-indazol-
4-
yloxy)-pro~an-1-one
CH3 O
/ ~ CH3 ~N I \
\ N /
HN ~O
N- O CHs
CH3
4M hydrochloric acid in dioxane (1 mL) was added to the compound of
Preparation 77 (45mg, 86pmol) in methanol and the solution was stirred for 2
hours. The reaction mixture was evaporated under reduced pressure and the
residue was dissolved in dichloromethane and basified with 1 M sodium
1o hydroxide solution to pHl2. The mixture was then passed through a phase
separation column and the filtrate was concentrated in vacuo. Purification by
column chromatography on silica gel, eluting with
dichloromethane:methano1:0.88 ammonia, 96:4:1, afforded the title
compound as a pale pin4c foam in 59~/~ yield, 20mg.
~ 5 LRMS (APCI+): m/z [M+H]+ 421
Example 87
l2Sl-1-f(2Rl-2-Methvl-4-lavridine-2-carbonyl)-aiaerazin-1-vll-2-(auinolin-5-
~iloxy~propan-1-one
O
/ I CH3 ~N \
N/ O N I /
\ O CH3
A mixture of the compound of Preparation 51 (150mg, 0.5mmol), picolinic
acid (49mg, 0.6mmol) O-(1H-benzotriazol-1-yl)-N,N,N',hP-tetramethyluronium
hexafluorophosphate (190mg, 0.75mmol) and triethylamine (0.93mL,
l0mmol) in dichloromethane (3mL) was stirred for 18 hours at room
temperature. The reaction mixture was then diluted with further
dichloromethane and washed with sodium hydroxide (lOmL) and water
(2x10mL). The organic phase was dried over magnesium sulfate and
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concentrated in vacuo. Purification of the residue by column chromatography-
on silica gel, eluting with dichloromethane:methanol, 99:1 to 95:5, afforded
the title compound as a white solid in 49% yield, 99mg.
LRMS (APCI+): m/z [M+H]+ 405
Example 88
(2S)-1-f4- 2-Fluoro-benzoyl)~2R)-2-methyl-loiperazin-1-girl]-2~(guinolin-5-
ylo~)-
propan-1-one
/ ~ CHs
N / ~ w0
\ O CH3
A mixture of the compound of Preparation 51 (150mg, 0.5mmol), 2-
fluorobenzoyl chloride and triethylamine in dichloromethane (3mL) was stirred
at room temperature for 18 hours. The reaction mixture was then diluted with
dichloromethane and washed with water (2x10mL). The organic phase was
dried over magnesium sulfate and concentrated in vacuo. Purification by
column chromatography on silica gel, eluting with dichloromethane:methanol,
98:2, afforded the title compound as a white solid in 73% yield, 153mg.
LRMS (APCI+): m/z [M+H]+ 422
2o Example 89
4-f (1 S)-2-f (2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy?-3-
methoxy-N-methyl-benzamide
O O
Me~H / Me ~N \
\~ N ~/
~O
OMe O Me
To a solution of (3R)-(3-methyl-piperazin-1-yl)-phenyl-methanone (J. Med.
Chem. (2000), 43(23), 4499) (22 g, 0.11 mol) and (2S)-2-(2-methoxy-4-
methylcarbamoyl-phenoxy)-propionic acid (25 g, 0.1 mol) (Preparation 5) in
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dry THF (250 ml) were added 1-N hydroxybenzatriazole monohydrate (20 g,
0.13 mol), 1-ethyl-3-(3'dimethylaminopropyl)carbodiimide.HCl (17 g, 0.09 mol)
and diisopropylethylamine (17 ml, 0.1 mol). After stirring for 1 hour a
further
portion of 1-ethyl-3-(3'dimethylaminopropyl)carbodiimide.HCl (17 g, 0.09 mol)
was added followed by additional diisopropylethylamine (17 ml, 0.1 mol).
After another hour a further portion of 1-ethyl-3-
(3'dimethylaminopropyl)carbodiimide.HCl (17 g, 0.09 mol) was added
followed by another portion of diisopropylethylamine (17 ml, 0.1 mol). The
mixture was then stirred for a further 16 hours. After this time LCMS analysis
1o indicated that all of the propionic acid had been consumed. The reaction
mixture was evaporated to give a viscous oil which was treated with water
(200 ml) followed by the slow addition of concentrated aqueous hydrochloric
acid until the mixture was pH 3. The resulting off-white precipitate was
filtered off and washed with water (3 x 100 ml). The solid was twice re-
crystallised from ethanol:water (1:1 by volume, 120 ml) to afford the title
compound as a white solid (32 g).
'H NMR (400MHz, CDC13): 8 : 7.50-7.30 (6H, m), 7.27 (1 H, m), 6.9 (1 H, m)
6.08 (1 H, br s), 5.1 (11 H, br m), 4.90-2.80 (7H, m), 3.90 (3H, br s), 3.00
(3H,
br s), 2.40 (3H, s), 1.70 (3H, br d), 1.40-0.9 (3H, m) ppm.
2o LRMS (ESI+) : m/z [M+H]+ 440
[a]p -26.9° (1 mg/ml in MeOH, 25°C, wavelength 589nM)
Example 90
5-((1 S~[(2R)-4-Benzoyl-2-meth~piperazin-1-~]-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid methyl ester
~ O
Me~O N
Me ~N ~ \
\ O N
OMe O Me
Di-tert-butyl azodicarboxylate (0.14 g, 0.6 mmol) and polymer supported
3o triphenylphosphine (0.25 g, 0.75 mmol) were added to a stirred solution of
(2R)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl)-2-hydroxy-propan-1-one
(Preparation 76) (0.083 g, 0.3 mmol) and 5-hydroxy-4-methoxy-pyridine-2-
carboxylic acid methyl ester (0.083 g, 0.45 mmol) (Tetrahedron Letters, 38,
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1297 (1997)) in dichloromethane (3 ml) at 0°C. After one hour the
reaction
mixture was allowed to warm to room temperature and additional di-tert-butyl
azodicarboXylate (0.14 g, 0.6 mmol) and polymer supported
triphenylphosphine (0.25 g, 0.75 mmol) were added. After 18 hours at room
5 temperature the reaction mixture was filtered through Arbocel~, washing
through with dichloromethane (10 mL). The filtrate was washed with
saturated aqueous sodium hydrogencarbonate solution (20 ml), then the
organic phase separated, dried (MgS04) and evaporated under reduced
pressure. Purification of the residue by column chromatography on silica gel,
1o eluting with ethyl acetate:pentane, 80:20, then ethyl acetate and finally
ethyl
acetate:methanol, 95:5, to afford the title compound as a white solid, 0.1 g.
LRMS (APCI+): m/z [M+H]+ 442
Example 91
15 5-{~1S -2-('(2R -4-BenzoVl-2-methyl-piperazin-1-yll-1-methyl-2-oxo-ethoxY}-
4-
methoxy-pyridine-2-carboxylic acid methylamide
O
Me
Me ~N ~ \
O N
O Me
2o A solution of 5-{(1S)-2-[(2R)-4-benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-
oxo-ethoxy}-4-methoxy-pyridine-2-carboxylic acid methyl ester (Example 90)
(0.2 g, 0.45 mmol) in a 33% w/w solution of methylamine in ethanol (3 ml)
was heated in a Reacti-vialT"" at 50°C for 18 hours. After cooling to
room
temperature solvent was evaporated under reduced pressure. The residue
25 was purified by column chromatography on silica gel, eluting with
dichloromethane:methanoI:NH3, 98:2:0.5 to 95:5:0.5, to afford the title
compound as a white solid, 0.18 g.
'H NMR (400MHz, CD30D): b : 8.05 (1 H, s), 7.75 (1 H, s), 7.50-7.40 (5H, m),
5.40 (1 H, m), 4.60-3.00 (8H, m), 3.95 (3H, br s), 2.90 (3H, s), 1.60-1.00
(6H,
so m) ppm.
Found C, 61.65; H, 6.47; N, 12.41. C23H2gN~O5. 0.1 mol CH2CI2 requires C,
61.80; H, 6.33; N, 12.48%
LRMS (APCI+): m/z [M+H]+ 441
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[a]o -15.5° (1 mg/ml in MeOH, 25°C, wavelength 589nM)
Example 92 ,
5-f ( 1 S)-2-j(2R)-4-Benzo~rl-2-methyl-~iperazi n-1-yl]-1-methyl-2-oxo-ethoxy)-
4-
methoxy-pyridine-2-carbox relic acid amide
0 0
H2N N ~ Me ~N ~ \
\ O N
OMe O Me
A solution of 5-{(1S)-2-[(2R)-4-benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-
oxo-ethoxy)-4-methoxy-pyridine-2-carboxylic acid methyl ester (Example 90)
(0.2 g, 0.45 mmol) in concentrated aqueous ammonia solution (2 ml) and
methanol (1 ml) was heated in a Reacti-vialT"' at 50°C for 18 hours.
After
pooling to room temperature solvent was evaporated under reduced pressure.
The residue was purified by column chromatography on silica gel, eluting with
dichloromethane:methanoI:NH3, 98:2:0.5 to 95:5:0.5, to afford the title
compound as a white solid, 0.16 g.
'H NMR (400MHz, CD3OD): 8 : 8.05 (1 H, s), 7.75 (1 H, s), 7.50-7.40 (5H, m),
5.40 (1 H, m), 4.60-3.00 (9H, m), 3.95 (3H, br s), 1.60-1.00 (6H, m) ppm.
Found C, 59.67; H, 6.11; N, 12.49. C22H26N4O5. 0.25 mol CH2C12 requires C,
59.78; H, 5.97; N, 12.54%
LRMS (APCI+): m/z [M+H]+ 427
Example 93
5-(~1S)-2-[(2R -4-Benzo~l-2-methyl-loiperazin-1-yl]-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid ethylamide
O O
Me~H N ( Me ~N \
\ O N U
I
OMe O Me
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A solution of 5-f (1 S)-2-[(2R)-4-benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-
-
oxo-ethoxy}-4-methoxy-pyridine-2-carboxylic acid methyl ester (Example 90)
(0.15 g, 0.34 mmol) in ethylamine (1 ml) was heated in a Reacti-vialT"" at
50°C for 18 hours. After cooling to room temperature solvent was
evaporated
under reduced pressure. The residue was purified by column
chromatography on silica gel, eluting with dichloromethane:methanol 96:4, to
afford the title compound as a white solid, 0.14 g.
'H NMR (400MHz, CD30D): b : 8.05 (1 H, s), 7.80 (1 H, s), 7.50-7.40 (5H, m),
5.40 (1 H, m), 4.60-3.00 (8H, m), 4.00 (3H, br s), 3.45 (2H, q), 1.60-1.00
(6H,
1o m), 1.20 (3H, t) ppm.
Found C, 60.89; H, 6.72; N, 11.93. C24H3aN4O5. 0.25 mol CH2C12 requires C,
61.22; H, 6.46; N, 11.78%
LRMS (APCI+): m/z [M+H]+ 455
Example 94
5-~(1 S)-2-f (2R)-4-Benzoyl-2-methyl-piperazin-1-~1-1-methyl-2-oxo-ethoxy}-4-
methoxy-pyridine-2-carboxylic acid cycloprop 1y amide
O
Me ~N \
N ~/
OMe O Me
A solution of 5-{(1S)-2-[(2R)-4-benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-
oxo-ethoxy}-4-methoxy-pyridine-2-carboxylic acid methyl ester (Example 90)
(0.12 g, 0.27 mmol) in cyclopropylamine (1 ml) was heated in a Reacti-vialT"'
at 50°C for 18 hours. After cooling to room temperature solvent was
2s evaporated under reduced pressure. The residue was purified by column
chromatography on silica gel, eluting with dichloromethane:methanol 95:5, to
afford the title compound as a white solid, 0.09 g.
'H NMR (400MHz, CD30D): b : 8.05 (1 H, s), 7.70 (1 H, s), 7.50-7.40 (5H, m),
5.40 (1 H, m), 4.60-3.00 (8H, m), 3.95 (3H, br s), 2.85 (1 H, m), 1.60-1.00
(6H,
3o m), 0.8 (2H, m), 0.6 (2H, m) ppm.
Found C, 63.02; H, 6.63; N, 11.83. C25H3pN4O5. 0.13 mol CH2C12 requires C,
63.20; H, 6.39; N, 11.73%
LRMS (APCI+): m/z [M+H]+ 467
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Preparation 1
~2S)-2- Quinolin-5-ylo~)-propionic acid methyl ester
Me
O OMe
Triphenylphosphine (2.17 g, 8.27 mmol) was added to a stirred suspension of
di-isopropyl azodicarboxylate (1.47 ml, 7.56 mmol) in THF (25 ml) at -
5°C.
5-Hydroxyquinoline (1.0 g, 6.90 mmol) and methyl (R)-lactate (0.66 ml, 6.90
1o mmol) were then added and the solution allowed to stir at room temperature
for 14 hours. The reaction mixture was diluted with ethyl acetate (50 ml),
washed with water (30 ml), saturated potassium carbonate solution (30 ml)
and brine (30 ml). The solution was dried (MgS04) and solvent evaporated
under reduced pressure. The crude product was purified by column
chromatography on silica gel eluting with a gradient system of pentane : ethyl
acetate (4:1 ) changing to pentane : ethyl acetate (3:2). The title compound
was obtained as a white solid (1.30 g).
LRMS (APCI+) : mlz [M+H]+ 232.
2o Preparation 2
(2S)-~Quinolin-5-yloxy)-propionic acid sodium salt
Me
~ O Na+
\ O
A solution of (S)-2-(quinolin-5-yloxy)-propionic acid methyl ester
(Preparation
1 ) (1.30 g, 5.60 mmol) and 1 N sodium hydroxide solution (5.62 ml) in dioxane
(40 ml) was heated at 60 °C for two hours. After this time tlc analysis
indicated that starting material remained so additional 1 N sodium hydroxide
solution (1.4 ml) was added and heating at 60 °C continued for one
hour.
so The reaction mixture was then concentrated under reduced pressure to afford
the title compound as a yellow solid which was used without further
purification.
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LRMS (ESI+) : m/z [M+H]+ 218.
PrJoaratiori 3
1-[(2R -4-Benzoyl-2-methyl-~i~erazin-1-yl]-2-bromo-propan-1-one
O
Me ~N ~ \
N
Br
O Me
To a solution of (3R)-(3-methyl-piperazin-1-yl)-phenyl-methanone (J. Med.
1o Chem. (2000), 43(23), 4499) (4.06 g, 19.6 mmol) in dichloromethane (60 ml)
and N,N dimethylformamide (2 ml) were added 2-bromopropionic acid (3.0 g,
19.6 mmol), O-(1 H-benzotriazol-1-yl)-N,N,I~P,N'-tetramethyiuronium
hexafluorophosphate (8.18 g, 21.6 mmol) and triethylamine (2.74 ml, 19.6
mmol). The reaction mixture was stirred at room temperature for 48 hours
and then washed with water (50 ml), 10 % w/v aqueous citric acid solution (50
ml) and saturated aqueous sodium hydrogencarbonate solution (50 ml). The
solution was dried (MgS~4) and solvent evaporated under reduced pressure.
The' crude product was purified by column chromatography on silica gel
eluting with a gradient system of dichloromethane: methanol (99:1 ) changing
2o to dichloromethane : methanol (95:5). The product was then purified further
by column chromatography on silica gel eluting with a gradient system of ethyl
acetate: dichloromethane (30:70) changing to ethyl acetate: dichloromethane
(45:55) to afford the title compound as a white solid (3.80 g).
LRMS (ESI+) : m/z [M+H]+ 340.
Preparation 4
4-Hydroxy-3-methoxy-N-methyl-benzamide
OH
Me
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To a mechanically stirred suspension of vanillic acid (84 g, 0.5 mol) in dry-
THF (300 ml) was added portion-wise carbonyldiimidazole (97 g, 0.6 mol).
Gaseous evolution was observed and the mixture became homogeneous
after approx 5 minutes. After further stirring for 30 minutes an off-white
5 precipitate form and stirring was maintained for a further 3 hours under
nitrogen. After this time LCMS analysis indicated that the vanillic acid had
been consumed and a solution of 2 M methylamine in THF (1000 ml, 2 mol)
was added in one portion and the mixture stirred for a further 16 hours to
generate a tan precipitate. The precipitate was filtered, washed with THF
o (2 x 250m1) followed by ether (300 ml) and dried under vacuum to afford the
N-methylammonium salt of the title compound (87g, 82%).
LRMS (ESI+): m/z [M+H]+ 182.
A portion of the ammonium salt {1.06g, 5 mmol) was refluxed in ethyl acetate
under a stream of nitrogen for 2 hours. The reaction was evaporated to give
~5 an oily solid, which was triturated with ethyl acetate to afford the title
compound as a tan solid (750 mg)
LRMS (ESI+): m/z [M+H]+ 182.
Preparation 5
20 (2S)-~2-Methox~4-methylcarbamoyl-phenoxy)-propionic acid
OH
Mew
To a stirred mixture of triphenylphosphine (42 g, 0.16 mol), 4-hydroxy-3-
25 methoxy-N-methyl-benzamide (Preparation 4) (20 g, 0.11 mol) and methyl
(R)-lactate (12 g, 0.12 mol) in dry THF (300 ml) at -5 °C was added
dropwise
a solution of di-isopropyl azodicarboxylate (32 ml, 0.16 mmol) in dry THF
(100 ml) over 30 minutes. The solution was allowed to warm to room
temperature and stirred for 2 hours. After this time tlc analysis indicated
that
3o the starting phenol had been consumed. The crude reaction mixture was
evaporated under reduced pressure to give a viscous oil. To a stirred solution
of this oil in methanol (150 ml) was added lithium hydroxide monohydrate (5.4
g, 0.13 mol) and the mixture stirred for 48 hours. After this time LCMS
analysis indicated that the intermediate methyl ester had been consumed.
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The crude reaction mixture was evaporated and partitioned between ether-
(500 ml) and water (150 ml). The aqueous phase was separated and washed
with ether (2 x 300 ml) and ethyl acetate (2 x 300m1). The aqueous phase
was then acidified to pH 2 using concentrated aqueous hydrochloric acid
resulting in the formation of a white precipitate. This precipitate was
filtered
off, washed with water (2 x 100m1) and recrystallised from ethanol water (1:1
by volume, 80 ml per 20 g) to afford the title compound as a white solid (21
g,
75%).
LRMS (ESI+): m/z [M+H]+ 254.
Preparation 6 ,
3-Methoxy-4-[(1 S)-1-methoxycarbon I-e~y]-benzoic acid methyl ester
H3C~0 CHs
O
O ~CH3
~O O
H3C
~iisopropyl azodicarboxylate (4mL, 20mmol) in tetrahydrofuran (l5mL) was
added dropwise to an ice-cooled solution of methyl vanillate (3.64g, 20mmol),
methyl (R)-lactate (2.08g, 20mmol) and triphenyl phosphine (5.24g, 20mmol)
in tetrahydrofuran (30mL) and the reaction mixture was stirred at room
temperature for 18 hours. The solvent was then evaporated under reduced
pressure and the residue was stirred in a mixture of diethyl ether (50mL) and
2o hexane (50mL). The resulting precipitate was filtered off and the filtrate
was
concentrated in vacuo. Purification of the residue by column chromatography
on silica gel, eluting with hexane:ethyl acetate, 85:15 to 75:25 afforded the
title compound in 71 % yield, 3.8g
LRMS (ES+): m/z [M+H]+ 269
30
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Preparation 7
4-[s1 S)-1-Carboxy-ethoxy]-3-methoxy-benzoic acid methyl ester
H3Cw0 CHs
OH
O
,O O
H3C
Lithium hydroxide monohydrate (4.2g, 100mmol) was added portionwise to a
solution of the compound of Preparation 6 (32g, 120mmol) in methanol
(150mL) and the mixture was stirred at room temperature for 18 hours. The
reaction mixture was then evaporated under reduced pressure and the
residue was dissolved in water and washed with diethyl ether. The aqueous
mixture was acidified with 2M hydrochloric acid to pH4 and was then
1o extracted with ethyl acetate (3x150mL). The combined organic extracts were
dried over magnesium sulfate and concentrated in vacuo. The resulting foam
was dried under reduced pressure to afford the title compound in 91 % yield,
23g.
LRMS (ES+): m/z [M+H]+ 255
Preparation 8
4-~~(1 S~-2-[(2R~4-Benzo~rl-2-methyl-piperazin-1-yll-1-methyl-2-oxo-ethoxy~-3-
methoxy-benzoic acid methyl ester
O
H~C~O CH3 ~N
N /
O
1
H CEO O CH3
3
2o Hianig's base (1.BmL, 10.5mmol) was added to a solution of the compound of
Preparation 7 (1.8g, 7mmol), (3R)-(3-methyl-piperazin-1-yl)-phenyl-
methanone [(1.6g, 7.7mmol), J. Med. Chem. 43(23), 4499; 2000], and 3-
(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (2.75g, 9.1 mmol) in
dichloromethane (l6mL). The mixture was stirred for 4 hours and was then
evaporated under reduced pressure. The residue was taken up in ethyl
acetate (200mL) and washed with 10% sodium carbonate solution (2x50mL)
and brine (2x50mL). The organic phase was dried over magnesium sulfate
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and concentrated in vacuo. Purification of the residue by column
chromatography on silica gel, eluting with hexane:ethyl acetate, 25:75 to 5:95
afforded the title compound in 95% yield, 2.95g.
LRMS (ES+): mlz [M+H]+ 441
Pr~aration 9
4~(1 S)-2-f(2R)-4-Benzoyl-2-methylpiperazin-1 yl1-1-methyl-2-oxo-ethoxy)-3-
methoxy-benzamide
H2N CH3
N
O
O
H3C
The compound of Preparation 8 (0.46g, 1.04mmol) and 2M ammonia in
methanol (lOmL) were heated in a sealed vessel at 120°C for 18 hours.
The
reaction mixture was then concentrated in vacuo and the residue was purified
by column chromatography on silica gel, eluting with
dichloromethane:methanol, 98:2 to 92:8, to afford the title compound as a
colourless foam in 55% yield, 243mg.
LRMS (ES+): m/z [M+H]+ 426
Preparation 10
4-f (1 S)-2-f (2R)-4-Benzoyl-2-meth r~l-piperazin-1-yll-1-methyl-2-oxo-
ethox~r)-N-
dimethylaminomethylene-3-methoxy-benzamide
H3C\N~CH3
~N CH
N
O
3
The compound of Preparation 9 (240mg, 0.56mmol) and N,N-
dimethylformamide dimethylacetal (lOmL) were heated under reflux for 7
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hours. The reaction mixture was then evaporated under reduced pressure to-
give a pale orange solid. The solid was purified by column chromatography
on silica gel, eluting with dichloromethane:methanol, 99:1 to 97:3, to provide
a
colourless oil. The oil was then azeotroped with dichloromethane and the
resulting foam was dried at 40°C for 3 hours to afford the title
compound in 94
yield, 255mg.
LRMS (ES+): m/z [M+Na]+ 503
Preparations 11 to 13
The following compounds, of the general formula shown below, were
prepared by the method described above for Preparation 10 using the
appropriate ketone and dimethylformamide dimethylacetal.
Rb
O
\ ~CH3
Ra
N~CH3
CH3
No. Ra Rb LRMS ES+ : m/z M+H +
11 CH3 H 220
12 H H 206
13 H OCH2-Ph 312
Preparation 11 uses 1-(3-methoxy-2-methyl-phenyl)-ethanone, prepared as
described in Tetrahedron 25(18), 4249; 1969.
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Preparatiori 14
3-(3-Methox~-2-meth rLl-phenyl)-1-methyl-1 H-pyrazole
5
\ OwCHs
~CH3
N
H3C
The compound of Preparation 11 (2.05g, 9.35mmol) was added portionwise
to a solution of methylhydrazine (547pL, 10.28mmol) in acetic acid (15mL)
1o and the mixture was heated at 90°C for 2.5 hours. The solvent was
then
evaporated under reduced pressure and the residue was dissolved in
dichloromethane and washed with 10% sodium carbonate solution and
saturated sodium hydrogen carbonate solution. The organic phase was dried
over magnesium sulfate and concentrated in vacuo to give an orange oil. The
15 oil was purified firstly by column chromatography on silica gel, eluting
with
dichloromethane:ethyl acetate, 99:1 to 97:3. This was followed by further
purification using an Isolute~ Flash silica column, eluting with
dichloromethane:ethyl acetate, 99:1 to 97:3, to afford the title compound as a
yellow oil in 31 % yield, 568mg.
2o LRMS (ES+): m/z [M+H]+ 203
Preparations 15 to 16
The following compounds, of the general formula shown below, were
25 prepared by the method described above for Preparation 14 using the
appropriate enamine and hydrazine monohydrate.
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Rb
O
\ ~CH3
/ Ra
H \
N
No. Ra Rb LRMS ES+ : m/z M+H +
15 H H 175
16 H OCH2Ph 281
s Preparation 17
2-Meth~(1-methyl-1 H ~yrazol-3-~~phenol
\ OH
~CH3
N ~~
N
H3C
Boron tribromide (1 M in dichloromethane, 8.16mL, 8.16mmol) was added
1o dropwise to an ice-cooled solution of the compound of Preparation 14
(550mg, 2.72mmol) in dichloromethane (6mL). The resulting solution was
allowed to warm to room temperature and was stirred for 2 hours. The
reaction mixture was then poured carefully onto ice and was neutralized with
10% sodium carbonate solution. The mixture was extracted with
1s dichloromethane (2x50mL) and the combined organic phases were dried over
magnesium sulfate and concentrated in vacuo. The residue was
chromatographed using an Isolute~ Flash silica column, eluting with
dichloromethane:methanol, 98:2 to 97:3, to give a brown oil. The oil was
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azeotroped with dichloromethane to afford the title compound as brown foam
in 61 % yield, 31 Omg.
LRMS (ESf): m/z [M+H]+ 189
Preparations 18 to 20
The following compounds, of the general formula shown below, were
prepared by the method described above for Preparation 17 using the
appropriate methoxybenzene derivative and boron tribromide.
NO. Ra R LRMS ES+ : m/z M+H
18 CH3 ~ 152
HaN
19 CH3 ~N 175
N\
20 H ~N 161
N'
Preparation 18: title compound is prepared by dealkylation of 3-methoxy-2-
methylbenzamide (J.Chem.Soc.Perkin Trans.l, 2389-2396; 1984)
Preparation 21
(2S)-2-[3-(2H-Pyrazol-3-yl~phenoxy]-propionic acid meth 1y ester
CHI
O
O ~CH3
O
H\~
N
OH
/
wRa
R°
Diisopropyl azodicarboxylate (295pL, 1.50mmol) was added dropwise to an
ice-cooled solution of the compound of Preparation 20 (240mg, l.5mmol)
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methyl (R)-lactate (156mg, 1.50mmol) and triphenyl phosphine (393mg;
l.5mmol) in tetrahydrofuran (5mL). The reaction mixture was stirred at room
temperature for 18 hours and was then diluted with diethyl ether (50mL) and
washed with water (2x10mL). The organic phase was dried over magnesium
sulfate and concentrated in vacuo to give a yellow oil. The oil was purified
on
an Isolute~ flash silica column, eluting with diethyl ether:dichloromethane,
80:20 to 95:5, to afford the title compound as a viscous yellow oil in 54%
yield, 200mg.
LRMS (ES-): m/z [M-H]- 245
Preaaration 22
~2S~j2-Methy~2H-pyrazol-3-yl)-phenoxyl-propionic acid methyl ester
CH3
O
O ~CH3
O
~CH3
H \
N
The title compound was prepared by the method described above for
Preparation 21 using the compound of Preparation 19 and methyl (R)-lactate:
LRMS (ES-): m/z [M-H]- 259
Preparation 23
(2S)-2-[3-(2H-Pyrazol-3- r1 -phenoxy~-propionic acid
CH3
OH
O
O
H ~
Lithium hydroxide monohydrate (80.9mg, 1.93mmol) was added portionwise
to a solution of the compound of Preparation 21 (190mg, 0.77mmol) in
methanol (6mL) and the mixture was stirred at 60°C for 8 hours. The
reaction
mixture was then evaporated under reduced pressure and the residue was
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dissolved in water and washed with diethyl ether. The aqueous mixture was
acidified with 2M hydrochloric acid to pH4 and was then extracted with ethyl
acetate (3x30mL). The combined organic extracts were dried over
magnesium sulfate and concentrated in vacuo. The resulting foam was dried
under reduced pressure to afford the title compound in 75% yield, 135mg.
LRMS (ES-): m/z [M-H]- 231
Preparation 24
!2S)-2-[2-Methyl-3- 2H-pyrazol-3-Lrl)-phenoxy]-propionic acid
CH3
OH
O
O
-CH3
H\
N
The title compound was prepared by the method described above for
Preparation 22 using the compound of Preparation 22 and lithium hydroxide.
LRMS (ES-): m/z [M-H]- 245
Preparation 25
1-[!2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-2-bromo-propan-1-one
0
CH3 ~N ~ \
N
Br
1
O CH3
2-Bromopropionic acid (1.53g, l0mmol) was added to a solution of N,N'-
dicyclohexylcarbodiimide (2.06g, l0mmol) in dichloromethane (40mL), cooled
to -15°C. The reaction mixture was stirred for 20 minutes before (2R)-
(3-
methyl-piperazin-1-yl)-phenyl-methanone [(2.04g, l0mmol), J. Med. Chem.
43(23), 4499; 2000] in dichloromethane (l6mL) was added dropwise. The
reaction mixture was stirred at -15°C for 90 minutes and was then
diluted with
diethyl ether (100mL) and stirred for a further 10 minutes. The mixture was
then filtered through Celite, washing through with diethyl ether and the
filtrate
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was concentrated in vacuo. The residue was purified by column
chromatography on silica gel, eluting with hexane:ethyl acetate, 40:60 to
20:80 to afford the title compound in 76% yield, 2.5g.
LRMS (ES+): m/z [M+H]+ 340
5
Preparation 26
3-~2-[(2R)-4-Benzoyl-2-methylpiperazin-1-yll-1-methyl-2-oxo-ethoxy~-2-
methyl-benzamide
O
/ CHs ~N ~ \
HEN \ N /
~O
O ~ ~ CHs
3
The compounds of Preparation 18 (180mg, 1.19mmol) and Preparation 25
(403.9mg, 1.19mmol) and cesium carbonate (387mg, 1.19mmol) in N,N-
dimethylformamide (lOmL) were heated at 80°C for 18 hours. The cooled
reaction mixture was then evaporated under reduced pressure and the
residue was partitioned between dichloromethane (50mL) and water (lOmL).
The organic phase was separated, dried over magnesium sulfate and
concentrated in vacuo. The residue was purified on an Isolute~ flash silica
column, eluting with dichloromethane:methanol, 99:1 to 96:4, to give a
colourless foam. The foam was azeotroped with dichloromethane and dried
2o under reduced pressure to afford the title compound in 98% yield, 475mg.
LRMS (ES+): m/z [M+H]+ 410
Preparation 27
1-[(2R~4-Benzoyl-2-meth-piperazin-1-~l-~3-methoxy-5-vitro-phenoxy)-
propan-1-one
Hs O
OzN / O CHs ~N
\I N U
O
O CHs
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The title compound was prepared by the method described above for -
Preparation 26 using the compound of Preparation 25 and 2-methoxy-5-
nitrophenol:
LRMS (APCI+): m/z [M+H]+ 428
Preparation 28
3-(2-f (2R)-4-Benzovl-2-methvlpiperazin-1-vll-1-methyl-2-oxoethoxvl-N
(dimethylamino~meth 1y ene]-2-methylbenzamide
i Hs
i
H C N ~ ~ CH3
N \ N
~O
p
3
The title compound was prepared by the method described above for
Preparation 10 using the compound of Preparation 26 and N,N-
dimethylformamide dimethylacetal.
LRMS (APCI+): m/z [M+H]+ 465
Preparation 29
3-~2-f (2R)-4-Benzovl-2-methyl-piperazin-1-vll-1-methyl-2-oxo-ethoxvl-N-
hydroxyaminomethylene-2-meth~rl-benzamide
H
HO~N~ ~ ~ GH3
IIN \ f
O
O CH3 O ,,
2o Sodium hydroxide solution (1 M, 1.06mL, 1.06mmol) was added dropwise to a
suspension of hydroxylamine hydrochloride (73.6mg, 1.06mmol) in acetic acid
(6mL). The compound of Preparation 28 (410mg, 0.88mmol) was then added
portionwise and the resulting solution was stirred at room temperature for 30
minutes. The solvent was evaporated under reduced pressure and the oily
residue was partitioned between dichloromethane (30mL) and water (6mL),
and neutralized with 10% sodium carbonate solution. The organic phase was
separated and the aqueous phase was re-extracted with dichloromethane
(30mL). The combined organic layers were dried over magnesium sulfate and
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concentrated in vacuo. The residue was purified on an Isolute~ flash silica
column, eluting with dichloromethane:methanol, 99:1 to 96:4, to afford the
title
compound as a colourless foam in 78% yield, 310mg.
LRMS (ES-): m/z [M-H]- 451
Preparation 30
~4-Amino-2-methoxy-phenoxy)-1-[(2R -4-benzoyl-2-methyl-piperazin-1=yll-
propan-1-one
Hs O
HaN / O CHs ~ N ~ \
\I N
0
O CH3
25% Pd/C (150mg) and ammonium formate (443mg, 7mmol) were added to a
solution of the compound of Preparation 27 (600mg, l.4mmol) in ethanol
(30mL) and the mixture was heated at 70°C for 2 hours. The reaction
mixture
was then cooled and filtered through Arbocel~, washing through with ethanol
(lOmL). The filtrate was concentrated in vacuo and the residue was purified
by column chromatography on silica gel, eluting with
dichloromethane:methanol, 95:5, to afford the title compound as a pale pink
solid in 80% yield, 443mg.
LRMS (APCI+): m/z [M+H]+ 398
Preparation 31
4-Benzylox~r-3-methoxy-benzamidine
H
"Butyl lithium (1.6M in hexane, 35.9mL, 57.47mmol) was added dropwise to a
solution of hexamethyldisilazane (11.64mL, 55.17mmol) in diethyl ether
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(60mL), cooled to -5°C, and the reaction mixture was stirred for 30
minutes
at a constant temperature of 0°C. 3-Methoxy-4-(phenylmethoxy)-
benzonitrile
(5.5g, 22.99mmol) was added portionwise and the mixture was stirred at
room temperature 4 hours. The reaction mixture was then poured onto ice
cold 2M hydrochloric acid (100mL) and was washed with diethyl ether (2x
100mL). The aqueous phase was basified to pHlO with 6M sodium hydroxide
solution and was extracted with dichloromethane (3x100mL). The combined
organic extracts were dried over magnesium sulfate and concentrated in
vacuo to afford the title compound as a white solid in 68% yield, 3.99g.
1o LRMS (ES+): m/z [M+H]+ 257
Preparation 32
2-(4-Benzyloxy-3-methoxy-~ohenyl)-1 H-imidazole
H
N
The compound of Preparation 31 (1 g, 3.90mmol) in chloroform (40mL) was
added to a solution of chloroacetaldehyde (336mg, 3.90mmol) in chloroform
(lOmL) and the mixture was stirred at room temperature for 18 hours. The
resulting white precipitate was filtered off and the filtrate was washed with
2o saturated sodium hydrogen carbonate solution (30mL). The aqueous phase
was re-extracted with 10% methanol in dichloromethane (2 x 50mL) and the
combined organic extracts were dried over magnesium sulfate and
concentrated in vacuo. The residue was purified on an Isolute~ flash silica
column, eluting with dichloromethane:methanol, 98:2 to 96:4, to give a brown
foam. The foam was purified further on an Isolute~ flash silica column,
eluting with dichloromethane:methanol, 99.5:0.5 to 98:2, to give a beige foam.
The foam was then azeotroped with dichloromethane and dried at
40°C to
afford the title compound in 16% yield, 180mg.
LRMS (ES+): m/z [M+H]+ 281
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Preparation 33
4J1 H-Imidazol-2-ail)-2-methoXy-phenol
OH
N ~ /
-O
~N CH3
To a solution of the compound of Preparation 32 (170mg, 0.61 mmol) in
methanol (lOmL) was added portionwise 10% Pd/C (85mg) and ammonium
formate (191 mg, 3.03mmol). The mixture was heated under reflux for 2
hours. The reaction mixture was then cooled and filtered through Arbocel~,
washing through with a mixture of dichloromethane and methanol (50:50,
250mL). The filtrate was concentrated in vacuo and the residue was purified
on an Isolute~ flash silica column, eluting with dichloromethane:methanol,
99:1 to 96:4, to afford the title compound as a white solid in 91 % yield,
105mg.
LRMS (ES+): mlz [M+H]+ 191
Preparation 34
2-Methoxy-4-(2H-pyrazol-3-yl~phenol
OH
N ~ /
Ni ~ ~ _O
CH3
The title compound was prepared by the method described above fior
Preparation 33 using the compound of Preparation 16.
LRMS (ES+): m/z [M+H]+ 191
Preparation 35
1-Benzyloxy-4-bromo-2-methoxy-benzene
Br \ / O
O
H3C
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A mixture of 4-bromo-2-methoxyphenol (5g, 24.63mmol), benzyl bromide-
(4.21 g, 24.63mmol) and cesium carbonate (8g, 24.63mmol) in acetone
(60mL) were heated under reflux for 4 hours. The cooled reaction mixture
was then evaporated under reduced pressure and the residue was partitioned
5 between diethyl ether (200mL) and water (80mL). The aqueous phase was
separated and re-extracted with diethyl ether (200mL). The combined organic
extracts were dried over magnesium sulfate and concentrated in vacuo. The
residue was azeotroped with dichloromethane to afford the title compound as
white solid in 100% yield, 7.198.
1 o LRMS (ES+): m/z [M+Na]+ 317
Preparation 36
1-(4-Benzyloxy-3-methoxy-phenyl)-1 H-pyrazole
N
~ \
N ~ ~ O
O
15 HsC
A mixture of the compound of Preparation 35 (1.5g, 5.12mmol), pyrazole
(697mg, 10.23mmol), copper (I) iodide (195mg, 1.02mmol) and potassium
carbonate in N,N-dimethylformamide (lOmL) was heated at 140°C for 18
hours. The cooled reaction mixture was then evaporated under reduced
2o pressure and the residue was partitioned between dichloromethane (50mL)
and water (30mL). The aqueous phase was separated and re-extracted with
diethyl ether (200mL). The combined organic extracts were dried over
magnesium sulfate and concentrated in vacuo. The residue was purified by
column chromatography on silica gel, eluting with dichloromethane:methanol,
25 100:0 to 96:4, to afford the title compound as a white solid in 82% yield,
1.18g.
LRMS (ES+): m/z [M+H]+ 281
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Preparation 37
2-Methoxy-4-~yrazol-1-yl-phenol
N
N ~ ~ OH
O
H3C
The title compound was prepared by the method described above for
Preparation 33 using the compound of Preparation 36.
LRMS (ES+): m/z [M+H]+ 191
Preparation 38
(2S)-~Quinolin-5-,~y~-propionic acid metal ester
CH3
O
N ~ ~ ~O ~CH3
\ O
Triphenylphosphine (2.17g, 8.27mmol) was added to a stirred suspension of
diisopropyl azodicarboxylate (1.47mL, 7.56mmol) in tetrahydrofuran (25mL) at
-5°C. 5-Hydroxyquinoline (1.0g, 6.90mmol) and methyl-(R)-lactate
(0.66mL,
6.90mmol) were added and the solution was allowed to stir at room
temperature for 14 hours. The reaction mixture was then diluted with ethyl
acetate (50mL), washed with water (30mL), saturated potassium carbonate
2o solution (30mL) and brine (30mL). The solution was dried over magnesium
sulfate and concentrated in vacuo. The residue was purified by column
chromatography on silica gel, eluting with pentane:ethyl acetate, 80:20 to
60:40, to afford the title compound as a white solid in 81 % yield, 1.30g.
LRMS (ES+): m/z [M+H]+ 232
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Preparation 39
(2S)-2-(Iso~uinolin-5-yloxy)-propionic acid meth~rl ester
/ I CH3
/ I O OwCHs
N\ O
The title compound was prepared by the method described above for
Preparation 38 using 5-hydroxyisoquinoline and methyl-(R)-lactate.
LRMS (APCI+): m/z [M+H]+ 232
Preparation 40
~2S)-2-(Quinolin-5-yloxy~-propionic acid sodium salt
I CH3
O Na+
N / I w0
O
A solution of the compound of Preparation 38 (1.308, 5.60mmol) and 1 M
sodium hydroxide solution (5.62mL) in 1,4-dioxane (40mL) was heated at
60°C for two hours. After this time tlc analysis indicated that
starting material
remained so additional 1 M sodium hydroxide solution (1.4m1) was added and
heating continued at 60°C for one hour. The reaction mixture was then
concentrated in vacuo to yield the title compound as a yellow solid in
2o quantitative yield.
LRMS (ES+): m/z [M+H]+ 218
Preparation 41
2S -2-~Isoguinolin-5-vloxv)-propionic acid sodium salt
I CH3
/ O Na+
N \ I _O
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The title compound was prepared by the method described above for
Preparation 40 using the compound of Preparation 39.
LRMS (APCI+): m/z [M+H]+ 218
Preparation 42
(2S)-1-f (2R)-4-Benzoyl-2-methyl-piperazin-1-Lrl]-~quinolin-5-yloxy)-propan-1-
one
0
/ I CH3 ~N I \
N/ I O N /
\ O CH3
1o A solution of the compound of Preparation 40 (1.0 g, 4.6mmol), (3R)-(3-
methyl-piperazin-1-yl)-phenyl-methanone [(0.94 g, 4.6mmol) J. Med. Chem.,
43(23), 4499; 2000], ~(1H-benzotriazol-1-yl)-N, N,N',N-tetramethyluronium
hexafluorophosphate (2.62g, 6.9mmol) and triethylamine (1.93m1, 13.8mmol)
in N,N dimethylformamide (30m1) was stirred at room temperature for 14
hours. The reaction mixture was then diluted with dichloromethane (100m1),
washed with water (3x50m1), dried over magnesium sulfate and concentrated
in vacuo. The residue was purified by column chromatography on silica gel
eluting with dichloromethane:methanol, 98:2, to afford the title compound as a
white solid in 49% yield, 0.91 g.
2o LRMS (ES+): m/z [M+H]+ 404
Preparation 43
(2S)-1-f(2R)-4-Benzoyl-2-methyl-piperazin-1-yl -~2-(isoguinolin-5-ylo~)-
propan-1-one
/ ~ CH3 ~N ~ \
O N /
N \ O CH3
The title compound was prepared by the method described above for
Preparation 42 using the compound of Preparation 41 and (3R)-(3-methyl-
piperazin-1-yl)-phenyl-methanone (J. Med. Chem., 43(23), 4499; 2000.
LRMS (APCI+): m/z [M+H]+ 404
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Preparation 44
(2S)-1-[~(2R~4-Benz~l-2-methyl-piperazin-1-yl]-2-(8-bromo-quinolin-5-yloxy)-
~ropan-1-one
Br
/ ~ CHI
N / ~ w0
J
A mixture of the compound of Preparation 42 (450mg, 1.1 mmol) and N-
bromosuccinimide (795mg, 4.4mmol) in acetonitrile (90mL) was stirred at
room temperature for 2 hours. The solvent was then evaporated under
1o reduced pressure and the residue was dissolved in ethyl acetate (50mL) and
washed with water (2x20mL). The organic phase was dried over magnesium
sulfate and concentrated in vacuo. The residue was purified by column
chromatography on silica gel, eluting with ethyl acetate:pentane, 33:67 to
100:0 to afford the title compound as a white solid in 98% yield, 526mg.
LFtMS (ES+): m/z [M+H]+ 482/484
Preparation 45
(2S)-1-f (2R)-4-Benzovl-2-methyl-piperazin-1-vl1-2-~[2-(2-trimethylsilan
etho~methy~-2H-~yrazol-3-yl]-quinolin-5-yloxy}-propan-1-one
H31
~ i\ CHs
N~N~O CH3
O
/ /
CH3 ~N ~ \
N/ O N /
O CH3
Tetrakis(triphenylphosphine)palladium(0) (6mg, cat), 1-[[(2-
(trimethylsilyl)ethoxy]methyl]pyrazolyl-5-boronic acid [(50mg, 0.1 mmol), J.
Med. Chem. 41 2019-2028; 1998] and sodium carbonate (175mg, l.6mmol)
in water (0.5mL) were added to the compound of Preparation 44 (50mg,
0.1 mmol) in tetrahydrofuran (2mL) and the mixture was heated under reflux
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for 18 hours. The reaction mixture was then evaporated under reduced
pressure and the residue was dissolved in dichloromethane and washed with
water (5m~) and brine (2x5mL). The organic phase was dried over
magnesium sulfate and concentrated in vacuo to give an orange oil. The oil
5 was purified by column chromatography on silica gel, eluting with
dichloromethane:methanol, 99:1 to 96:4 to afford the title compound as a
yellow solid.
LRMS (APCI+): m/z [M+H]+ 600
1 o Preparation 46
~2S)-1-[(2R)-4-Benzoyl-2-methyl-piperazin-1-yl]-~1-oxy-quinolin-5- I~oxy)-
prop~an-1-one
CH3
O~N+i O
\ ~ O
meta-Chloroperbenzoic acid (1.54g, 4.44mmol) was added to an ice-cooled
solution of the compound of Preparation 42 (1.5g, 3.7mmol) in
dichloromethane (l5mL) and the mixture was stirred at room temperature for
3 hours. The reaction mixture was then diluted with dichloromethane (lOmL)
2o and washed with saturated aqueous potassium carbonate solution (lOmL)
and water (2x 10m1). The organic layer was dried over magnesium sulfate
and concentrated in vacuo to afford the title compound as a white solid in
quantitative yield, 1.~g.
LRMS (APCI+): m/z [M+H]+ 420
30
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Pre~~aration 47
5-f2-f 2R -4-Benzoyl-2-methyl-piperazin-1-yl]-1-methyl-2-oxo-ethoxy~-
quinoline-2-carboxylic acid meth, 1y ester
O
CH3 ~N ~ \
N
N ~ ~ ~O
O \ O CH3
O
H3C~
The chloro compound of Preparation 66 (130mg, 0.3mmol), dichloro-
bis(triphenylphosphine)palladium (24mg, 0.03mmol) and triethylamine (83pL,
0.6mmol) were dissolved in a mixture of methanol (3mL) and N,N-
dimethylformamide (0.5mL), and transferred to a sealed vessel. The vessel
1o was heated to 100°C and the mixture was stirred under 100psi of
carbon
monoxide gas for 42 hours. The reaction mixture was then filtered through
Arbocel~, washing through with methanol and the filtrate was concentrated in
vacu~. Purification of the residue by column chromatography on silica gel,
eluting with ethyl acetate:pentane, 75:25 to 100:0, afforded the title
compound in 41 % yield, 50mg.
LRMS (APCI+): m/z [M+H]+ 462
Preparation 48
5-~2-f f 2 R)-4-Benzoyl-2-methyl-piperazin-1-y~-1-methyl-2-oxo-ethox~r}-
2o quinoline-2-carboxylic acid
OH
N
1 M Sodium hydroxide solution (130pL, 0.135mmol) was added to a solution
of the compound of Preparation 47 (40mg, 0.09mmol) in dioxane (3mL) and
the mixture was heated at 60°C for 3 hours. The solvent was then
evaporated
under reduced pressure and the residue was dissolved in water (20mL) and
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washed with ethyl acetate. The phases were separated and the organic
phase was discarded. The aqueous phase was acidified with glacial acetic
acid and re-extracted with ethyl acetate. The retained organic phase (2"d
washing) was dried over magnesium sulfate and concentrated in vacuo to
afford the title compound as a pale yellow solid in 67% yield, 26mg.
LRMS (APCI+): m/z [M+H]+ 448
Preparation 49
(2Sl-1-f(2Rl-4-Benzovl-2-methyl-aiperazin-1-vl1-2-(2-benzvlamino-auinolin-5-
1 o ylox~r -propan-1-one
O
CH3 ~N ~ \
N
\ O
O CH3
N /
NH
/
A mixture of the compound of Example 60 (50mg, 0.11 mmol), tetrabutyl
ammonium fluoride (30mg, 0.22mmol), triethylamine (0.16mL, 1.1 mmol) and
benzylamine (0.13mL, 1.1 mmol) in dimethylsulfoxide (1 mL) was heated at
120°C for 72 hours. The reaction mixture was then purified directly by
HPLC
using a Phenomenex Luna C18 system, eluting with
water/acetonitrile/trifluoroacetic acid (5:95:0.1 ):acetonitrile, 95:5 to
5:95, to
afford the title comloound in 34% yield.
LRMS (APCI+): m/z [M+H]+ 509
25
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Preparation 50
(3R)-3-Methyl-4-[(2S~2-(quiriolin-5-ylox~i -propionyl]-piperazine-1-carboxylic
acid tert-but, 1y ester
O CHs
/ CH N~O CH3
3
CH3
N
N ~ ~ ~O
\ 0 CH3
1-Hydroxybenzotriazole hydrate (249mg, 1.87mmol) was added to a
suspension of the compound of Preparation 40 (400mg, l.7mmol) in N,N-
dimethylformamide to form a solution. 1-Ethyl-3-(3-dimethyl amino propyl)
carbodiimide (351 mg, 1.87mmol) and triethylamine (0.7mL, 5.1 mmol) were
1o added and the solution was stirred for 5 minutes. (3S)-3-Methyl-piperazine-
1
carboxylic acid tent butyl ester (335 mg, 1.7mmol) was added and the mixture
was stirred for 18 hours. The solvent was then evaporated under reduced
pressure and the residue was dissolved in ethyl acetate (30mL) and washed
with water (20mL). The organic phase was dried over magnesium sulfate and
~5 concentrated in vacuo to afford the title compound in 48% yield, 332mg.
LRMS (APCI+): m/z [M+H]+ 400
Preparation 51
(2S)-1-[(2R)-2-Methyl-piperazin-1-yll-2-(quinolin-5-yloxy)-propan-1-one;
2o hydrochloride
HCI
CH3 ~N.
N
N ~ ~ ~O
\ O CH3
The compound of Preparation 50 (300mg, 0.75mmol) was stirred in 4M
hydrochloric acid in dioxane (3mL) for 2 hours. The reaction mixture was
concentrated in vacuo to afford the title compound as a pale yellow solid in
25 quantitative yield, 348 mg.
LRMS (APCI+): mlz [M+H]+ 300
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Preparation 52
(2S)-1-[(2R)-4-Benzo~l-2-meth rLl-piperazin-1-yll-2-(2,8-dichloro-auinolin-5-
ylox~~-propan-1-one
CH3
t
O
O CH3
A mixture of the compound of Example 60 (200mg, 0.4mmol) and N-
chlorosuccinimide (61 mg, 0.4mmol) in acetonitrile (1 OmL) was heated at
40°C
for 48 hours. The solvent was then evaporated under reduced pressure and
o the residue was dissolved in dichloromethane (20mL) and washed with water
(2x10mL). The organic phase was dried over magnesium sulfate and
concentrated in vacuo to give a yellow oil. The residue was purified by column
chromatography on silica gel, eluting with dichloromethane:methanol, 98:2, to
afford the title compound as a white solid in 78% yield, 169mg.
LRMS (APCI+): m/z [M+H]+ 472
Preparation 53
~2S~ 1-[(2R)-2-MethLrl-4-_(pyridine-2-carbonyl)-piperazin-1-yll-2-(auinolin-5-
ylox~r)-propan-1-one
O
~ I CH3 ~N \
,0 1
\ O CH3
A mixture of the compound of Preparation 51 (150mg, 0.5mmol), picolinic
acid (49mg, 0.6mmol) O-(1 H-benzotriazol-1-yl)-N,N,I~P,N-tetramethyluronium
hexafluorophosphate (190mg, 0.75mmol) and triethylamine (0.93mL,
l0mmol) in dichloromethane (3mL) was stirred for 18 hours at room
temperature. The reaction mixture was then diluted with further
dichloromethane and washed with sodium hydroxide (lOmL) and water
(2x10mL). The organic phase was dried over magnesium sulfate and
concentrated in vacuo. Purification of the residue by column chromatography
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on silica gel, eluting with dichloromethane:methanol, 99:1 to 95:5, afforded
the title compound as a white solid in 49% yield, 99mg.
LRMS (APCI+): m/z [M+H]+ 405
5 Preparation 54
j2S)-1-[4-~(2-Fluoro-benzoyl)-(2R)-2-methyl-piperazin-1-yl]-2~quinolin-5-
yloxy)-
pro~an-1-one
CH3
N / ~ w0
\ O CH3
1o A mixture of the compound of Preparation 51 (150mg, 0.5mmol), 2-
fluorobenzoyl chloride and triethylamine in dichloromethane (3mL) was stirred
at room temperature for 18 hours. The reaction mixture was then diluted with
dichloromethane and washed with water (2x10mL). The organic phase was
dried over magnesium sulfate and concentrated in vacuo. Purification by
15 column chromatography on silica gel, eluting with dichloromethane:methanol,
98:2, afforded the title compound as a white solid in 73% yield, 153mg.
LRMS (APCI+): m/z [M+H]+ 422
Preparation 55
20 4-Bromo-2-trifluoromethoxy-phenol
OH
O eF
F F
Br
Bromine (449mg, 2.81 mmol) was added to a solution of 2-
(trifluoromethoxy)phenol (500mg, 2.81 mmol) and sodium acetate (169mg,
25 2.81 mmol) in acetic acid (5mL) and the solution was stirred for 1 hour.
The
reaction mixture was then diluted with water (30mL) and extracted with ethyl
acetate (2x50mL). The organic extracts were combined, dried over
magnesium sulfate and concentrated in vacuo to give a colourless oil. The oil
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was purified by column chromatography on silica gel, eluting with -
pentane:ethyl acetate, 85:15, to afford the title compound as a white solid in
23% yield.
LRMS (APCI+): m/z [M+H]+ 255/257
Preparation 56
~2S)-2- 4-Bromo-2-trifluoromethoxy-phenoxy~propionic acid methyl ester
H3C O-CH3
O
F
1o Diisopropyl azodicarboxylate (414pL, 2.14mmol) was added dropwise to an
ice-cooled solution of the compound of Preparation 55 (500mg, 1.95mmol)
methyl (R)-lactate (202mg, 1.95mmol) and triphenyl phosphine (614mg,
2.34mmol) in tetrahydrofuran (20mL). The reaction mixture was stirred at
room temperature for 18 hours and was then evaporated under reduced
pressure. The residue was dissolved in ethyl acetate and washed with 10%
potassium carbonate solution (2x10mL). The organic phase was dried over
magnesium sulfate and concentrated in vacuo. The residue was purified by
column chromatography on silica gel, eluting with dichloromethane:methanol,
95:5, to afford the title compound as a colourless oil in 76% yield, 638mg.
2o LRMS (APCI+): m/z [M+H]+ 305/307
Preparation 57
S)-2-(4-Bromo-2-trifluoromethoxy-phenoxy)-propionic acid
H3C OH
g O
A mixture of the compound of Preparation 56 (574mg, 1.67mmol) and 1 M
sodium hydroxide solution (4.2mL, 4.2mmol) in dioxane (lOmL) was stirred at
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55°C for 3 hours, The solvent was then evaporated under reduced
pressure
and the residue was dissolved in water (30mL). The aqueous solution was
acidified to pH2 with 2M hydrochloric acid and extracted with ethyl acetate
(2x50mL). The combined organic extracts were dried over magnesium sulfate
and concentrated in vacuo to afford the title compound as a yellow solid in
80% yield.
LRMS (APCI+): mlz [M+H]+ 327
Preparation 58
(2S~-1-[S2R)-4-Benzoyl-2-methyl-piherazin-1-yl]-2-(4-bromo-2-
trifluoromethoxy-phenoxy)-propan-1-one
0
CH3 ~N
N /
O CH3
F
A solution of the compound of Preparation 57 (150mg, 0.46mmol), (3Fi)-(3-
methyl-piperazin-1-yl)-phenyl-methanone [(93mg, 0.46mmol) J. Med. Ghem.,
43(23), 4499; 2000], O-(1H-benzotriazol-1-yl)-N,N,N,hP-tetramethyluronium
hexafluorophosphate (260mg, 0.69mmol) and triethylamine (0.13m1,
0.92mmol) in dichloromethane (30m1) was stirred at room temperature for 18
hours. The reaction mixture was then diluted with dichloromethane (50m1)
2o and washed with water (50m1). The aqueous phase was re-extracted with
dichforomethane (50mL) and the combined organic extracts were dried over
magnesium sulfate and concentrated in vacu~. The residue was purified on
an Isolute~ flash silica column, eluting with dichloromethane:methanof, 99:1,
to afford the title compound as a white foam in 90% yield, 213mg.
LRMS (APCI+): m/z [M+H]+ 5371539
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Preparation 59
4-,;~1 S)-2-[(2R -4-Benzoyl-2-methyl-piperazin-
1-girl]-1-methyl-2-oxo-ethoxy~-3-trifluoromethoxy-benzoic acid methyl ester
O
H3Cw0 ~N ~ \
N /
CH3
The compound of Preparation 58 (190mg, 0.37mmol), dichloro-
bis(triphenylphosphine)palladium (30mg, 0.04mmol) and triethylamine
(0.10mL, 0.74mmol) were dissolved in methanol (lOmL) and transferred to a
o sealed vessel. The vessel was heated to 100°C and the mixture was
stirred
under 100psi of carbon monoxide gas for 42 hours. The reaction mixture was
then filtered through Arbocel~, washing through with methanol and the filtrate
was concentrated in ~acu~. The residue was purified on an Isolute~ flash
silica column, eluting with dichloromethane:methanol, 97:3 to afford the title
compound as an orange solid in 85% yield, 155mg.
LRMS (APCI+): m/z [M+H]+ 495
Preparations 60 to 61
2o The following compounds, of the general formula shown below, were
prepared by the method described above for Preparation 26 using the
compound of Preparation 25 and the appropriate hydroxyquinoline or
hydroxyisoquinoline.
O
CH3 ~N ~ \
R\ N /
O
O CH3
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No. R5 LRMS APCI+ : m/z M+H +
60 404
\ \
N
61 404
\ \
N / /
The following compounds, of the general formula shown below, were
prepared by the method described above for Preparation 46 using the
s compounds of Preparations 60 and 61 and meta-chloroperbenzoic acid
0
CH3 ~N ( \
R~ N /
O
O CH3
No. R5 LRMS APCI+ : m/z M+H +
62 420
\ \
N+
(_
O
63 420
\ \
- iN / /
O
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Preparatiori 64
(2S;I-1-f(2R-4-Benzoyl-2-methyl-piperazin-1-yl]-~1-ox -~~uinolin-5- lyoxy)~
5 propan-1-one
CH3
N
O
\ ~ J
- O~N / /
The title compound was prepared by the method described above for
1o Preparation 46 using the compound of Preparation 43.
LRMS (APCI+): m/z [M+H]+ 420
Preparation 65
~2S)-1-[(2R)-4-Benzoyl-2-meth rL1-piperazin-1-yl]-~1-chloro-isoquinolin-5-
15 yloxK -propan-1-one
o
CH3 ~N ~ \
N /
O
\ \ O CH3
N / /
CI
Phosphorus oxychloride (0.20mL, 2.18mmol) was added to a solution of the
compound of Preparation 64 (305mg, 0.73mmol) in dichloromethane (l5mL)
and the reaction mixture was heated at 100°C for two hours. The mixture
was
2o then cooled to room temperature and poured onto water (75mL). The
resulting aqueous mixture was basified with concentrated ammonia solution
and extracted with dichloromethane (2x75mL). The combined organic
extracts were then dried over magnesium sulfate and concentrated in vacuo.
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Purification of the residue by column chromatography on silica gel, eluting -
with dichloromethane:methanol, 99:1, afforded the title compound as a white
foam in 40% yield, 127mg.
LRMS (APCI+): m/z [M+H]+ 438
Preparation 66
1-[f 2R)-4-Benz~l-2-methyl-piperazin-1-yl]-2-(2-chloro-quinolin-5-~y)-
propan-1-one
O
CH3 ~N
N
O
O CH3
CI N
1o The title compound was prepared by the method described above for
Preparation 65 using the compound of Preparation 62.
LRMS (APCI+): m/z [M+H]+ 438
Preparation 67
2-Methoxy-4-methylsulfanyl-phenol
H3C
OH
tButyl lithium (1.7M in pentane, 14.5mL, 24.6mmol) was added dropwise to a
solution of 4-bromo-2-methoxyphenol (2.00g, 9.85mmol) in tetrahydrofuran
(25mL) cooled to -78°C. The mixture was stirred for 15 minutes and was
then
warmed to -40°C and stirred for a further 30 minutes. Dimethyl
disulfide
(1.06mL, 11.8mmol) was added and the mixture was stirred at room
temperature for 18 hours. Water was then added to the reaction and the
resulting mixture was acidified to pH1 with 2M hydrochloric acid. The aqueous
phase was separated and re-extracted with ethyl acetate. The combined
organic fractions were then dried over magnesium sulfate and concentrated in
vacuo to give an orange oil. This oil was purified by column chromatography
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on silica gel, eluting with pentane:diethyl ether, 95:5 to 80:20, to afford
the
title compound as a white solid in 48% yield, 800mg.
'HNMR(CDC13, 400MHz) S: 2.40(s, 3H), 3.90(s, 3H), 5.50(brs, 1 H), 6.80-
6.90(m, 3H)
Prea~aration 68
6-Hydroxy-7-methox -y 3,4-dihydro-2H-isoquinolin-1-one
OH
HN ~ ~ OMe
O
A mixture of lithium iodide (0.32g, 2.41 mmol) and 6,7-dimethoxy-3,4-dihydro-
2H-isoquinolin-1-one (0.5 g, 2.41mmol) in 2,4,6-collidine (10m1) was heated at
130°C for 18 hours. The solvent was then evaporated under reduced
pressure and the residue was purified by column chromatography on silica
gel, eluting with dichloromethane:methanol, 96:4, to afford the title
com~aound
as a pale yellow solid, 0.19 g
LRMS (APCI+): m/z [M+H]+ 194
Preparation 69
2o Acetic acid 3-acetoxv-5-methyl-ahenvl ester
0
O~CH
3
H3C O
H C_ 'O
3
Acetic anhydride (22.6mL, 0.24mo1) was added to an ice-cooled solution of
3,5-dihydroxytoluene (9.93g, 0.08mo1) and triethylamine (56mL, 0.40mo1) in
dichloromethane (86mL). The reaction mixture was warmed to room
temperature and was stirred for 60 hours. Water (100mL) was then added
and the mixture was stirred vigorously for 3 hours. The organic layer was
separated and the aqueous layer was re-extracted with dichloromethane
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(3x80mL). The combined organic extracts were washed with brine (80mL);
dried over magnesium sulfate and concentrated in vacuo. The residue was
purified by column chromatography on silica gel, eluting with ethyl
acetate:pentane, 66:33, to afford the title compound as a colourless oil in
94% yield, 15.7g.
LRMS (APCI+): m/z [M+NH4]+ 226
Preparation 70
~2.6-Dihydroxy-4-methyl-phenyl)-ethanone
OH O
\ ~CH3
HsC OH
A solution of the compound of Preparation 69 (10.12g, 48.6mmol) in
chlorobenzene (lOmL) was added dropwise to a suspension of aluminium
chloride (19.44g, 145.8mmol) in chlorobenzene (50mL), warmed to 90°C,
and
the mixture was stirred for one hour. The reaction mixture was then cooled
and was pipetted carefully onto a mixture of ice and 2M hydrochloric acid.
The resulting mixture was extracted with ethyl acetate (3x200mL) and the
combined organic layers were washed with brine, dried over magnesium
sulfate and concentrated in vacuo. The residue was purified by column
chromatography on silica gel, eluting with pentane:ethyl acetate, 84:16,
80:20, 75:25 to afford the title compound as a yellow solid in 66% yield,
5.31 g.
LRMS (APCI+): m/z [M+H]+ 167
Preparation 71
3,8-Dimethyl-1 H-indazol-4-0l
OH CH3
~\
/
HsC H
A solution of the compound of Preparation 70 (4.15g, 25mmol) in ethylene
glycol (70mL) was added dropwise to a solution of hydrazine monohydrate
so (2.4mL, 50mmol) in ethylene glycol (l5mL) and the mixture was stirred at
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room temperature for 1 hour and at 150°C for 80 minutes. The cooled
reaction mixture was then poured onto water and was acidified to pH6 with
acetic acid. The aqueous mixture was extracted with ethyl acetate (4x200mL)
and the combined extracts were washed with 5% sodium sulphite (200mL)
and brine (200mL). The organic phase was dried over magnesium sulfate and
concentrated in vacuo. Purification of the residue by column chromatography
on silica gel, eluting with ethyl acetate:pentane, 50:50, afforded the title
compound as a yellow solid in 95% yield, 3.84g.
LRMS (APCI+): m/z [M+H]+ 163
Preparation 72
4-~tert-Butyl-dimethyl-silanLrlox~r)-3,6-dimethyl-1 H-indazole
~ Hs CHs
H3C- i i CHs
CHs
N_. CHs
tent Butyldimethylchlorosilane (511 mg, 3.39mmol) and imidazole (1.05g,
15.4mmol) were added to a solution of the compound of Preparation 71
(500mg, 3.08mmol) in N,N-dimethylformamide and the reaction mixture was
stirred at 0°C for 1 hour and at room temperature for 18 hours.
Additional tent
2o butyldimethylchlorosilane (511 mg, 3.39mmol) was added and the reaction
mixture was heated at 80°C for 48 hours. The solvent was then
evaporated
under reduced pressure and the residue was partitioned between ethyl
acetate and water. The organic phase was separated, washed with brine
(2x10mL), dried with magnesium sulfate and concentrated in vacuo. The
residue was purified by column chromatography on silica gel, eluting with
pentane:ethyl acetate, 15:85, to afford the title compound as an orange oil in
23% yield.
LRMS (APCI+ : m/z [M+Hl+ 27
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Preparation 73
4- tart-Butyl-dimethyl-silanyloxy)-3,6-dimethyl-indazole-1-carboxylic acid
tart
butyl ester
i ~3 CH3
H3C- I i~CH3
H3C ~ O ~CH3
/~.cH3
N-N
O
H3C ~~~
H3C CH3
O
Di-tart-butyl dicarbonate (503mg, 2.3mmol) and 4-dimethylaminopyridine
(51 mg, 0.4mmol) were added to the compound of Preparation 72 (580mg,
2mmol) in dichloromethane (5mL) and the reaction mixture was stirred for 18
1o hours. Water was then added to the reaction mixture and the aqueous
mixture was extracted with dichloromethane (3x10mL). The combined organic
extracts was dried over sodium sulfate and concentrated in vacuo.
Purification of the residue by column chromatography on silica gel, eluting
with pentane:ethyl acetate, 95:5, afforded the title compound as a colourless
oil in 66% yield, 527mg.
LRMS (APCI+): m/z [M+HJ+ 377
Pr~aration 74
4-Hydroxy-3,6-dimethyl-indazole-1-carboxylic acid tart-but I e,~ster
H3C ~ OH
~CH3
N_N
O
H3C \\~
O
HsC CHs
Tetrabutylammonium fluoride solution (1 M in tetrahydrofuran, 21 mL,
2.1 mmol) was added dropwise to a solution of the compound of Preparation
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73 (527mg, 1.4mmol) in tetrahydrofuran (9mL) and the solution was stirred at
0°C for 30 minutes and at room temperature for 5 minutes. Water was
then
added to tre mixture and the aqueous mixture was extracted with ethyl
acetate (2x10mL). The combined organic extracts were washed with brine,
dried over magnesium sulfate and concentrated in vacuo. Purification by
column chromatography on silica gel, eluting with pentane:ethyl acetate,
70:30 to 60:40, afforded the title compound as a white solid in 76% yield,
280mg.
LRMS (APCI-): mlz [M-H]- 261
Preparation 75
(2R)-1-[~,2R)-4-Benzoyl-2-meth r~l-piperazin-1-~]-2-benzyloxy-propan-1-one
O
H3C\
N
O ~N \
H3C
\ O
1-Hydroxybenzotriazole hydrate (825mg, 5.3mmol), N-methylmorpholine
(808pL, 7.3mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (1.13g, 5.8mmol) and (3R)-(3-methyl-piperazin-1-yl)-phenyl-
methanone [(1.0g, 4.9mmol), J. Med. Chem. 43(23), 4499; 2000] were added
to a solution of (R)-(+)-2-benzyloxypropionic acid (972mg, 5.3mmol) in
2o dichloromethane (lOmL) and the mixture was stirred for 18 hours. Additional
(R)-(+)-2-benzyloxypropionic acid (486mg, 2.65mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.65mg, 2.9mmol),
1-hydroxybenzotriazole hydrate (413mg, 2.65mmol) and N-methylmorpholine
(808pL, 7.3mmol) were added and the mixture was heated under reflux for 3
2s hours. Water was then added to the reaction mixture and the aqueous
solution was extracted with dichloromethane (2x10mL). The combined
organic extracts were washed with 2M hydrochloric acid, 1 M sodium
hydroxide solution and brine. The organic phase was dried over magnesium
sulfate and concentrated in vacuo. Purification by column chromatography on
3o silica gel, eluting with pentane:ethyl acetate, 50:50, followed by ethyl
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acetate:methanol, 90:10, afforded the title compound as a white foam in 81
yield, 1.45g
LRMS (APCI-): m/z [M-H]- 367
Preparation 76
(2R)-1-[(2R)-4-Benzoyl-2-meth~rl-piperazin-1-yl -2-hydrox~propan-1-one
O
H3C
- N~ ~
OH ~N
H3C~' ~
O
Pd(OH)2 (300mg, 2.2mmol) and ammonium formate (1.37g, 22mmol) were
1o added to a solution of the compound of Preparation 75 (1.458, 3.9mmol) in
ethanol (30mL) and the mixture was heated at 60°C for 2 hours. Tlc
analysis
then showed that the reaction had not reached completion so additional
Pd(OH)2 (300mg, 2.2mmol) was added, followed by ammonium formate
(1.37g, 22mmol) added at 45 minute intervals until all of the starting
material
was consumed. The reaction mixture was then cooled and filtered through
Arbocel~, washing through with ethanol (lOmL). The filtrate was concentrated
in vacuo and the residue was purified by column chromatography on silica
gel, eluting with dichloromethane:methanol, 95:5, to afford the title compound
as a white gum in 91 % yield, 1 g.
2o LRMS (APCI+): m/z [M+H]+ 277
Preparation 77
4-[(,1 S~2-[~2R)-4-Benzo~rl-2-methyl-piperazin-1-arl]-1-methyl-2-oxo-ethoxyl-
3,6-dimethyl-indazole-1-carboxylic acid terl-butyl ester
CH3 O
CH3
H3C~O ~ CH3 N \
H3C ~ \ _ N
O N O
N- O CHs
CH3
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The compound of Preparation 76 (70mg, 0.25mmol), di-tert-butyl
azodicarboxylate (233mg, 1.01 mmol) and polymer supported triphenyl
phosphine (380mg, 1.l3mmol) were added to a solution of the compound of
Preparation 74 (70mg, 0.25mmol) in dichloromethane (3mL) and the reaction
mixture was stirred 0°C for 30 minutes and at room temperature for 18
hours.
The reaction mixture was then filtered through a filter tube, washing through
with dichloromethane. The filtrate was washed with sodium hydroxide
solution, dried over magnesium sulfate and concentrated in vacuo.
Purification of the residue by column chromatography on silica gel, eluting
1o with pentane:ethyl acetate, 40:60 to 20:80 afforded the title compound as a
white foam in 34% yield, 45mg
LRMS (APCI+): m/z [M+H]+ 521
Biological Data
The ability of the compounds of formula (I) and their pharmaceutically
acceptable salts, solvates and derivatives to modulate gp120 activity, in
particular inhibit the interaction of gp120 with CD4, is demonstrated using a
gp160 induced cell-cell fusion assay to determine the ICSO values of
2o compounds against HIV-1 fusion. The gp160 induced cell-cell fusion assay
uses a HeLa P4 cell line and a CHO-TatlO cell line.
The HeLa P4 cell line expresses CCR5 and CD4 and has been
transfected with HIV-1 LTR-(3-Galactosidase. The media for this cell line is
Dulbecco modified eagle's medium (D-MEM) (without L-glutamine) containing
10% foetal calf serum (FCS), 2mM L-glutamine penicillin/streptomycin
(Pen/Strep; 100U/mL penicillin + l0mg/mL streptomycin), and 1 p.g/ml
puromycin.
The CHO cell line is a Tat (transcriptional traps activator)-expressing
clone from a CHO JRR17.1 cell line that has been transfected with prat puro
so plasmid. The media for this cell line is rich medium for mammalian cell
culture originally developed at Roswell Park Memorial Institute RPM11640
(without L-glutamine) containing 10% FCS, 2mM L-glutamine, 0.5 mg/ml
Hygromycin B and 12pg/ml puromycin. The CHO JRR17.1 line expresses
gp160 (JRFL) and is a clone that has been selected for its ability to fuse
with
a CCRS/CD4 expressing cell line.
Upon cell fusion, Tat present in the CHO cell is able to transactivate
the HIV-1 long terminal repeat (LTR) present in the HeLa cell leading to the
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expression of the (3-Galactosidase enzyme. This expression is then
measured using a Fluor AceT"" ~i-Galactosidase reporter assay kit (Bio-Rad
cat no. 170-3150). This kit is a quantitative fluorescent assay that
determines
the level of expression of ~i-galactosidase using 4-methylumbelliferul-
galactopyranoside (MUG) as substrate. (i-Galactosidase hydrolyses the
fluorogenic substrate resulting in release of the fluorescent molecule
4-methylumbelliferone (4MU). Fluorescence of 4-methylumbelliferone is then
measured on a fluorometer using an excitation wavelength of 360nm and
emission wavelength of 460nm.
1o Compounds that inhibit fusion will give rise to a reduced signal and,
following solubilisation in an appropriate solvent and dilution in culture
medium, a dose-response curve for each compound can be used to calculate
ICSO values.
All the Examples of the invention have ICSO values, according to the
above method, of less than 1.SpM. ICSO values for the compounds of
Examples 12, 29 and 44 are, respectively, l5nM, 134nM and 825nM.
The ability of compounds of formula (I) to inhibit the interaction of
gp120 with CD4 is further demonstrated using an enzyme linked
immunosorbent assay (ELISA). Maxisorp plates (Nunc) are coated with
2~.g/well of anti-gp120 antibody (D7324). 100p,1 of gp120 (dilution pre-
determined by titration) is added to each well and incubated for 90 minutes at
room temperature. The sample is removed and the wells are washed with
PBS (phosphate buffered saline) + 0.01% TWEEN~ (polyethylene glycol
sorbitan monolaurate). 50~1/well of compound is added followed by 50p,1
(0.1 ~.g) of soluble CD4 conjugated to horseradish peroxidase (Autogen
Bioclear). The plate is incubated for 90 minutes at room temperature before
the wells are washed again. The substrate ~PD (o-phenylenediamine,
Sigma) is added at a concentration of 0.5mg/ml and the plate incubated in the
dark at room temperature for 3 minutes before 3M HCI is added to stop the
3o reaction. Compounds that inhibit the interaction of gp120 with soluble CD4
will give rise to a reduced absorbance at 492nm.
All the Examples of the invention have ICSO values, according to the
above method, of less than 15~.M. ICSO values for the compounds of
Examples 1, 11 and 12 are, respectively, 0.92p,M, 1.Op,M and 0.75p,M.
