Note: Descriptions are shown in the official language in which they were submitted.
CA 02225366 1997-12-19
HOECHST AKTIENGESELLSCHAFT HOE 961F 354 Dr.EKlsch
Description
Substituted purine derivatives, processes for their preparation, their use,
and
compositions comprising them
The present invention relates to compounds of the formulae I and la
W Wa
N N N
\ ~~Y ~N i ~~Y
N / _N N
X N ~ X
G (I) Ga (la)
in which X, Y, W, Vila, G and Ga have the meanings indicated below, and to
their
physiologically tolerable salts and their prodrugs, their preparation, their
use and
pharmaceutical preparations comprising them.
The compounds of the formula I are valuable pharmaceutical active compounds.
In
particular, they are vitronectin receptor antagonists and are suitable for the
therapy
and prophylaxis of illnesses which are based on the interaction between
vitronectin
receptors and their ligands in cell-cell or cell-matrix interaction processes
or which
can be prevented, alleviated or cured by influencing this interaction. The
invention
relates, inter alia, to the use of compounds of the formula I and of their
physiologically tolerable salts and of pharmaceutical preparations which
contain
such compounds, as therapeutics for the prevention, alleviation or cure of
illnesses
which are caused at least partially by an undesired extent of bone resorption,
angiogenesis or proliferation of cells of the vascular smooth musculature, or
for
whose therapy or prophylaxis an influencing of these processes is intended. In
particular, the compounds of the formula I are suitable, for example, as
inhibitors of
bone resorption, as inhibitors of tumor growth and tumor metastasis, as
antiinflammatories, for the treatment or prophylaxis of cardiovascular
disorders,
CA 02225366 1997-12-19
2
such as, for example, arteriosclerosis or restenosis, or for the treatment or
prophylaxis of nephropathies and retinopathies, such as, for example, diabetic
retinopathy.
The compounds of the formulae I and la according to the invention inhibit bone
resorption by osteoclasts. Bone diseases against which the compounds of the
formula I can be employed are especially osteoporosis, hypercalcemia,
osteopenia,
for example caused by metastases, dental disorders, hyperparathyroidism,
periarticular erosions in rheumatoid arthritis and Paget's disease. The
compounds
of the formula I can furthermore be employed for the alleviation, avoidance or
therapy of bone disorders which are caused by glucocorticoid, steroid or
corticosteroid therapy or by a lack of sex hormone(s). All these disorders are
characterized by bone loss which is based on the inequilibrium between bone
formation and bone destruction.
Human bones are subject to a constant dynamic renovation process comprising
bone resorption and bone formation. These processes are controlled by types of
cell
specialized for these purposes. Bone formation is based on the deposition of
bone
matrix by osteoblasts, and bone resorption is based on the destruction of bone
matrix by osteoclasts. The majority of bone disorders are based on a disturbed
equilibrium between bone formation and bone resorption. Osteoporosis is
characterized by a loss of bone matrix. Activated osteoclasts are polynuclear
cells
having a diameter of up to 400 Nm, which remove bone matrix. Activated
osteoclasts
become attached to the surface of the bone matrix and secrete proteolytic
enzymes
and acids into the so-called "sealing zone", the region between their cell
membrane
and the bone matrix. The acidic environment and the proteases cause the
destruction of the bone.
Studies have shown that the attachment of osteoclasts to the bones is
controlled by
integrin receptors on the cell surface of osteoclasts. Integrins are a
superfamily of
receptors which include, inter alia, the fibrinogen receptor a~~bf33 on the
blood
platelets and the vitronectin receptor a"f33. The vitronectin receptor a~f33
is a
membrane glycoprotein which is expressed on the cell surface of a number of
cells
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3
such as endothelial cells, cells of the vascular smooth musculature,
osteoclasts and
tumor cells. The vitronectin receptor a~f33, which is expressed on the
osteoclast
membrane, controls the process of attachment to the bone and bone resorption
and
thus contributes to osteoporosis. a~f33 in this case binds to bone matrix
proteins
such as osteopontin, bone sialoprotein and thrombospontin, which contain the
tripeptide motif Arg-Gly-Asp (or RGD).
Norton and co-workers describe RGD peptides and an anti-vitronectin receptor
antibody (23C6) which inhibit tooth destruction by osteoclasts and the
migration of
osteoclasts (Norton et al., Exp. Cell. Res. 1991, 195, 368). In J. Cell Biol.
1990, 111,
1713, Sato et al. describe echistatin, an RGD peptide from snake venom, as a
potent inhibitor of bone resorption in a tissue culture and as an inhibitor of
osteoclast adhesion to the bone. Fischer et al. (Endocrinology 1993, 132, 1411
)
were able to show in the rat that echistatin also inhibits bone resorption in
vivo.
Wayne et al. (J. Clin. Invest. 1997, 99, 2284) were able to demonstrate in the
rat the
in vivo efficacy of the inhibition of bone resorption by a vitronectin
receptor
antagonist.
The vitronectin receptor a~f33 on human cells of the vascular smooth
musculature of
the aorta stimulates the migration of these cells into the neointima, which
finally
leads to arteriosclerosis and restenosis after angioplasty (Brown et al.,
Cardiovascular Res. 1994, 28, 1815).
Brooks et al. (Cell 1994, 79, 1157; J. Clin. Invest. 96 (1995) 1815) and
Mitjans et al.,
J. Cell Science 1995, 108, 2825) showed that antibodies against a~f33 or a~f33
antagonists can cause a shrinkage of tumors by inducing the apoptosis of blood
vessel cells during angiogenesis. Cheresh et al. (Science 1995, 270, 1500)
describe
anti-a~f33 antibodies or a~f33 antagonists which inhibit the bFGF-induced
angiogenesis process in the rat eye, a property which can be used
therapeutically in
the treatment of retinopathies.
EP-A-0 528 586 and EP-A-0 528 587 disclose aminoalkyl- or heterocyclyl-
substituted phenylalanine derivatives, and WO 95/32710 discloses aryl
derivatives
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as inhibitors of bone resorption by osteoclasts. WO 95128426 describes RGD
peptides as inhibitors of bone resorption, angiogenesis and restenosis. WO
96/00574 and WO 96126190 describe benzodiazepines, inter alia, as vitronectin
receptor antagonists or integrin receptor antagonists. WO 96/00730 describes
fibrinogen receptor antagonist templates, in particular benzodiazepines, which
are
linked to a nitrogen-bearing 5-membered ring, as vitronectin receptor
antagonists.
EP-A-0 531 883 describes fused 5-membered heterocycles which inhibit
fibrinogen
binding to platelets.
The present invention relates to compounds of the formulae I and la
W Wa
N ~ N
~N , \~Y ~N ~ ~~Y
/ _N N / _N N
X ~ X
G (I) Ga (la)
in which:
X is hydrogen, NRsRs~, fluorine, chlorine, bromine, ORs, SRs, hydroxy-(C~-Cs)-
alkyl-NH, (hydroxy-(C~-Cs)-alkyl)2N, amino-(C~-Cs)-alkyl-NH, (amino-(C~-Cs)-
alkyl)2N, hydroxy-(C~-C6)-alkyl-O, hydroxy-(C~-C6)-alkyl-S or NH-CO-Rs;
Y is Rs, fluorine, chlorine, bromine, cyano, NRsRs~, ORs, SR6 or hydroxy-(C~-
Cs)-alkyl-NH;
G is a radical of the formula II
-(CR~R2)~-A-(CR~R2)m-(CR~R3)~ (CR~R2)q R4 (II);
W is a radical of the formula III
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-B-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-D-E (III);
Ga is a radical of the formula Ila
5 -(CR1 R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-D-E (11a);
Wa is a radical of the formula Illa
-B-(CR~R2)~-A-(CR~R2)m-(CR~R3)i (CR~R2)q R4 (Illa);
A, A' independently of one another are a direct bond, -C(O)NR5-, -NRSC(O)-,
-C(O)-, -NR5-, -O- , -S-, -SO-, -S02-, (C5-C~4)-arylene, where in the aryl
radical one to five carbon atoms can be replaced by one to five heteroatoms,
(C2-C4)-alkynylene, (C2-C4)-alkenylene, or a divalent radical of a 3- to 7-
membered saturated or unsaturated ring which can contain one or two
heteroatoms, such as, for example, nitrogen, sulfur or oxygen, and which can
be monosubstituted or disubstituted by radicals from the group consisting of
=0, =S and R3;
R~, R2 independently of one another are hydrogen, fluorine, chlorine, cyano,
nitro,
(C~-C~o)-alkyl, (C3-C~4)-cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (CS-
C14)-aryl, (C5-C~4)-arYl-(C~-C8)-alkyl, R6-O-R~, Rs-S(O)p-R7 or R6Rs~N-R7;
R3 independently of one another is hydrogen, fluorine, chlorine, cyano, nitro,
(C~-C~8)-alkyl, (C3-C~4)-cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-
C~4)-a~'Yl, (C5-C~4)-arYl-(C~-C8)-alkyl, Rs-O-R7, R6R6~N-R~, R6C(O)-0-R7,
RsC(O)R7, R60C(0)R~, RsN(Rs~)C(O)OR~, R6S(O)pN(R5)R~,
R60C(O)N(R5)R7, RsC(O)N(RS)R~, R6N(Rs~)C(O)N(R5)R~,
R6N(R6~)S(O)pN(R5)R~, R6S(O)PR~, RsSC(O)N(R5)R7, RsN(Rs~)C(O)R~ or
RsN(Rs~)S(O)PR~, where alkyl can be monounsaturated or polyunsaturated
and where furthermore alkyl and aryl can be monosubstituted or
polysubstituted by fluorine, chlorine, bromine, cyano, R6Rs~NR~, nitro,
RsOC(O)R~, RsC(O)R~, RsN(Rs~)C(O)R~, RsN(Rs~)S(O)PR~, Rs or Rs-O-R~;
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R4 is C(O)R8, C(S)R8, S(O)PR8, P(O)R$R8~ or a radical of a four- to eight-
membered saturated or unsaturated heterocycle which contains 1, 2, 3 or 4
heteroatoms from the group consisting of N, O, S, such as, for example,
tetrazolyl, imidazolyl, pyrazolyl, oxazolyl, thiadiazolyl;
RS independently of one another is hydrogen, (C~-C~o)-alkyl, (C3-C~4)-
cycloalkyl,
(C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl or (C5-C~4)-aryl-(C~-C8)-
alkyl;
R6, R6~ independently of one another are hydrogen, (C~-C~8)-alkyl, (C3-C~4)-
cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, in which 1 to 5
carbon atoms can be replaced by heteroatoms such as N, O, S, or (C5-C~4)-
aryl-(C~-Cg)-alkyl, in which, in the aryl moiety, 1 to 5 carbon atoms can be
replaced by heteroatoms such as N, 0, S, or Rs and Rs~ , together with the
atoms connecting them, form a ring system, in particular a 4- to 8-membered
ring system which can optionally also contain additional, in particular one,
two or three additional, heteroatoms from the group consisting of N, O, S
and which can be saturated or unsaturated, in particular saturated, such as,
for example, morpholine, thiomorpholine, piperazine, piperidine, pyrrolidine;
R7 independently of one another is (C~-C4)-alkylene or a direct bond;
R8, R8~ independently of one another are hydroxyl, (C~-C$)-alkoxy, (C5-C~4)-
aryl-
(C~-C8)-alkoxy, (CS-C~4)-aryloxy, (C~-C8)-alkylcarbonyloxy-(C~-C4)-alkoxy,
(C5-C~4)-aryl-(C~-C8)-alkylcarbonyloxy-(C~-C8)-alkoxy, NRsRs~, (di((C~-C8)-
alkyl)amino)carbonylmethyloxy, (di((C5-C~4)-aryl-(C~-C8)-alkyl)-
amino)carbonylmethyloxy, (C5-C~4)-arylamino, the radical of an amino acid,
N-((C~-C4)-alkyl)piperidin-4-yloxy, 2-methylsulfonylethoxy, 1,3-thiazol-2-
ylmethyloxy, 3-pyridylmethyloxy, 2-(di((C~-C4)-alkyl)amino)ethoxy or the
radical Q- (CH3)3N+-CH2-CH2-O-, in which Q- is a physiologically tolerable
anion;
B is -O-, -S-, -NR5-, -NR5-C(O)-, -C(O)-NR5-, a direct bond or a divalent
radical of a 3- to 7-membered saturated or unsaturated ring which can
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7
contain one or two heteroatoms, such as, for example, nitrogen, sulfur or
oxygen, and which can be monosubstituted or disubstituted by radicals from
the group consisting of =O, =S and R3;
D is a direct bond, -NRs-, -C(O)-NRs-, -NRs-C(O)-, -S(0)~-NRs-,
-NRs-C(O)-NR6-, -NRs-C(S)-NRs-, -NRs-S(O)S NR6-, -NRs-C(O)O-,
-NR6-N=CRs-~ -NR6-S(O)~ , -(C5-C~4)-arYl-CO- , -(C5-C~4)-arYl-S(0)~ ,
-N=CR6-, -RsC=N- or -R6C=N-NRs-, where the divalent radicals
representing D are bonded to the group E via the free bond on the right side;
E is hydrogen, R6-C(=NR6)-NR6-, R6Rs~N-C(=NR6)-, R6R6~N-C(=NRs)-NR6- or
a radical of a 4- to 11-membered, monocyclic or polycyclic, aromatic or
nonaromatic ring system which can optionally contain 1, 2, 3 or 4
heteroatoms from the group consisting of N, O and S and can optionally be
monosubstituted, disubstituted or trisubstituted by radicals from the group
consisting of R3, R5, =O, =S and R6R6~N-C(=NRs)-, such as, for example,
the following radicals:
3 5
N R R R5
N ~ R3 N Ra N
N .
15
R N N
Rs Rs
I ~ S R3 R3 S
N
R
()
N
N N , N ,
0
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H
R3 ~ ~ ~ ~ Ra
\ ~ ~ \ N
N \ HN 1 ~ N
~ H Rs
HN ' _ NH
z
/ N / ( )v / 1 Iv / ( w
R3 w ~ N/ Rs \ ~ ~ Ra ~N~ ~ Ra \
N ~5 N N N ~ N
R . Rs , R6 ; R5 ,
H Rs
N /
R3 \ I ~ N
N , N I ~ \
H R5 R5 , N N N
R5
R5
Rs Rs
N N N~N
N II
N N ~ I / N~N
N_N ~N ,
1 5
R
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N N ~ N ~ N\
R5
0 R O ~N~N ~N N '
R5
. O
R5 NH2
N
N
N N
NH I
N
I H2N N ,
0 ,
5
n is zero, one, two, three, four or five;
m is zero, one, two, three, four or five;
i is zero or one;
p independently of one another is zero,
one or two;
q is zero, one or two;
r is zero, one, two, three, four, five
or six;
s is zero, one, two, three, four or five;
t is zero, one, two, three, four or five;
k is zero or one;
a is one or two;
v is zero, one, two or three;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts and their prodrugs;
where, instead of the purine structure shown in the formulae I and la, a
3-deazapurine structure, a 7-deazapurine structure or a 7-deaza-8-azapurine
structure can also be present.
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All radicals and indices which can occur several times in the compounds of the
formulae I and la, for example the radicals R~, R2 and R3 occurring in the
radicals G
and W and the radicals R5, R6, Rs~, R~ and indices occurring therein, but also
all
other radicals and indices to which this applies, can each independently of
one
5 another have the meanings indicated. They can be identical or different.
Likewise,
heteroatoms in heterocycles or substituents in radicals which can be present
several
times independently of one another can have the meanings indicated and can be
identical or different.
10 The alkyl radicals occurring in the substituents can be straight-chain or
branched,
saturated or mono- or polyunsaturated. This also applies if they carry
substituents or
occur as substituents of other radicals, for example in alkoxy radicals,
alkoxycarbonyl radicals or aralkyl radicals. The same applies to the divalent
alkylene radicals.
Examples of suitable (C~-C~8)-alkyl radicals are: methyl, ethyl, propyl,
butyl, pentyl,
hexyl, heptyl, octyl, decyl, undecyl, dodecyl, tridecyl, hexadecyl, octadecyl,
the
n-isomers of these radicals, isopropyl, isobutyl, isopentyl, neopentyl,
isohexyl,
3-methylpentyl, 2,3,4-trimethylhexyl, sec-butyl, tart-butyl, tert-pentyl.
Preferred alkyl
radicals are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl
and tert-
butyl.
Unsaturated alkyl radicals are, for example, alkenyl radicals such as vinyl, 1-
propenyl, allyl, butenyl, 3-methyl-2-butenyl or alkynyl radicals such as
ethynyl, 1-
propynyl or propargyl. Alkenylene and alkynylene radicals can be straight-
chain or
branched. Examples of alkenylene radicals are vinylene or propenylene,
examples
of alkynylene radicals are ethynylene or propynylene.
Cycloalkyl radicals can be monocyclic, bicyclic or tricyclic. Monocyclic
cycloalkyl
radicals are, in particular, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl and cyclooctyl, furthermore, for example, cyclononyl, cyclodecyl,
cycloundecyl, cyclododecyl or cyclotetradecyl, which, however, can also all be
substituted by, for example, (C~-C4)-alkyl. Examples of substituted cycloalkyl
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11
radicals which may be mentioned are 4-methylcyclohexyl and
2, 3-dimethylcyclopentyl.
Bicyclic and tricyclic cycloalkyl radicals can be unsubstituted or can be
substituted
in any desired suitable positions by one or more oxo groups andlor one or more
identical or different (C~-C4)-alkyl groups, for example methyl groups or
isopropyl
groups, preferably methyl groups. The free bond of the bicyclic or tricyclic
radical
can be located in any desired position of the molecule; the radical can thus
be
bonded via a bridgehead atom or via an atom in a bridge. The free bond can
also be
located in any desired stereochemical position, for example in an exo-position
or an
endo-position.
Examples of parent structures of bicyclic ring systems are norbornane
(= bicyclo[2.2.1 ]heptane), bicyclo[2.2.2]octane and bicyclo[3.2.1 ]octane. An
example
of a system substituted by an oxo group is camphor
(= 1,7,7-trimethyl-2-oxobicyclo[2.2.1 ]heptane).
Examples of parent structures of tricyclic systems are twistane
(= tricyclo[4.4Ø03'8]decane, adamantane (= tricyclo[3.3.1.13~~]decane),
noradamantane (= tricyclo[3.3.1.03~~]nonane), tricyclo[2.2.1.02'6]heptane,
tricyclo[5.3.2.04'9]dodecane, tricyclo[5.4Ø02°9]undecane or
tricyclo[5.5.1.03~~ ~ ]tridecane.
(CS-C~4)-aryl includes heterocyclic (C5-C~4)-aryl radicals in which ring
carbon atoms
are replaced by heteroatoms such as nitrogen, oxygen or sulfur, and
carbocyclic
(C6-C~4)-aryl radicals. Examples of carbocyclic aryl radicals are phenyl,
naphthyl,
biphenylyl, anthryl or fluorenyl, where 1-naphthyl, 2-naphthyl and in
particular
phenyl are preferred. Aryl radicals, in particular phenyl radicals, can be
monosubstituted or polysubstituted, preferably monosubstituted, disubstituted
or
trisubstituted, by identical or different radicals from the group consisting
of (C~-C8)-
alkyl, in particular (C~-C4)-alkyl, (C~-C8)-alkoxy, in particular (C~-C4)-
alkoxy,
halogen, such as fluorine, chlorine and bromine, nitro, amino,
trifluoromethyl,
hydroxyl, methylenedioxy, cyano, hydroxycarbonyl, aminocarbonyl, (C~-C4)-
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12
alkoxycarbonyl, phenyl, phenoxy, benzyl, benzyloxy, (R90)2P(O)-, (R90)2P(O)-O-
or
tetrazolyl, where R9 is hydrogen, (C~-C~o)-alkyl, (Cs-C~4)-aryl or (Cs-C~4)-
aryl-
(C~-C8)-alkyl. The same applies to the corresponding arylene radicals.
In monosubstituted phenyl radicals, the substituent can be located in the 2-
position,
the 3-position or the 4-position, the 3- and the 4-position being preferred.
If phenyl
is disubstituted, the substituents can be in the 1,2-, 1,3- or 1,4-position
relative to
one another. With respect to the linkage site, the substituents can be located
in the
2,3-position, 2,4-position, 2,5-position, 2,6-position, 3,4-position or 3,5-
position.
Preferably, in disubstituted phenyl radicals the two substituents are arranged
in the
3-position and the 4-position, relative to the linkage site.
Aryl groups or arylene groups can also be monocyclic or polycyclic aromatic
ring
systems in which 1, 2, 3, 4 or 5 ring carbon atoms are replaced by
heteroatoms,
such as, for example, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrrolyl, furyl,
thienyl, imidazolyl,
pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, tetrazolyl, pyridyl,
pyrazinyl,
pyrimidinyl, indolyl, isoindolyl, indazolyl, phthalazinyl, quinolyl,
isoquinolyl,
quinoxalinyl, quinazolinyl, cinnolinyl, f3-carbolinyl, or a benzo-fused,
cyclopenta-,
cyclohexa- or cyclohepta-fused derivative of these radicals. These
heterocycles can
be substituted by the same substituents as the abovementioned carbocyclic aryl
systems.
In the series of these aryl groups or of the corresponding arylene groups,
monocyclic or bicyclic aromatic ring systems having 1, 2 or 3 heteroatoms from
the
group consisting of N, O, S, which can be unsubstituted or substituted by 1, 2
or 3
substituents from the group consisting of (C~-C6)-alkyl, (C~-Cs)-alkoxy,
fluorine,
chlorine, nitro, amino, trifluoromethyl, hydroxyl, (C~-C4)-alkoxycarbonyl,
phenyl,
phenoxy, benzyloxy and benzyl, are preferred.
Particularly preferred here are monocyclic or bicyclic aromatic 5- to 10-
membered
ring systems having 1 to 3 heteroatoms from the group consisting of N, O, S,
which
can be substituted by 1 to 2 substituents from the group consisting of (C~-C4)-
alkyl,
(C~-C4)-alkoxy, phenyl, phenoxy, benzyl and benzyloxy.
CA 02225366 1997-12-19
1'
V
Examples of saturated and unsaturated rings, in particular of 3- to 7-membered
saturated or unsaturated rings which can contain one or two heteroatoms such
as,
for example, nitrogen, sulfur or oxygen and can optionally be monosubstituted
or
disubstituted by =O, =S or R3, are cyclopropane, cyclobutane, cyclopentane,
cyclohexane, cycloheptane, cyclopentene, cyclohexene, cycloheptene,
tetrahydropyran, 1,4-dioxacyclohexane, morpholine, thiomorpholine, piperazine,
piperidine, pyrrolidine, dihydroisoxazole, tetrahydroisoxazole, 1,3-dioxolane,
1,2-dithiolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran,
2,3-dihydrothiophene, 2,5-dihydrothiophene, 2-imidazoline, 3-imidazoline,
4-imidazoline, 2-oxazoline, 3-oxazoline, 4-oxazoline, 2-thiazoline, 3-
thiazoline,
4-thiazoline, thiazolidine, a-thiapyran, a-pyran, y-pyran.
The radical of an amino acid representing R8 andlor R8~ is, as usual in
peptide
chemistry, formally obtained by removing a hydrogen atom from the amino group
of
the amino acid. By means of the free bond on the amino group formally obtained
hereby, the radical of the amino acid is then bonded, for example, to the CO
group
in the group CO-R8. Amino acids can be natural or unnatural amino acids. a-
Amino
acids are preferred. Amino acids can exist in different stereochemical forms,
for
example as D- or L-amino acids, and in stereochemically homogeneous form or in
the form of mixtures of stereoisomers. Amino acids which may be mentioned, for
example, are (cf. Houben-Weyl, Methoden der organischen Chemie [Methods of
Organic Chemistry], Volume XVI1 and 2, Georg Thieme Verlag, Stuttgart, 1974):
Aad, Abu, ~yAbu, ABz, 2ABz, EAca, Ach, Acp, Adpd, Ahb, Aib, (iAib, Ala, f3Ala,
DAIa,
Alg, All, Ama, Amt, Ape, Apm, Apr, Arg, Asn, Asp, Asu, Aze, Azi, Bai, Bph,
Can, Cit,
Cys, (Cys)2, Cyta, Daad, Dab, Dadd, Dap, Dapm, Dasu, Djen, Dpa, Dtc, Fel, Gln,
Glu, Gly, Guv, hAla, hArg, hCys, hGln, hGlu, His, hlle, hLeu, hLys, hMet,
hPhe,
hero, hSer, hThr, hTrp, hTyr, Hyl, Hyp, 3Hyp, Ile, Ise, Iva, Kyn, Lant, Lcn,
Leu, Lsg,
Lys, f3Lys, eLys, Met, Mim, Min, nArg, Nle, Nva, Oly, Orn, Pan, Pec, Pen, Phe,
Phg,
Pic, Pro, Pro, Pse, Pya, Pyr, Pza, Qin, Ros, Sar, Sec, Sem, Ser, Thi, f3Thi,
Thr,
Thy, Thx, Tia, Tle, Tly, Trp, Trta, Tyr, Val, tert-butylglycine (Tbg),
neopentylglycine
(Npg), cyclohexylglycine (Chg), cyclohexylalanine (Cha), 2-thienylalanine
(Thia),
CA 02225366 1997-12-19
14
2,2-diphenylaminoacetic acid, 2-(p-tolyl)-2-phenylaminoacetic acid, 2-(p-
chlorophenyl)aminoacetic acid;
furthermore:
pyrrolidine-2-carboxylic acid; piperidine-2-carboxylic acid; 1,2,3,4-
tetrahydroisoquinoline-3-carboxylic acid; decahydroisoquinoline-3-carboxylic
acid;
octahydroindole-2-carboxylic acid; decahydroquinoline-2-carboxylic acid;
octahydrocyclopenta(b]pyrrole-2-carboxylic acid; 2-azabicyclo[2.2.2]octane-3-
carboxylic acid; 2-azabicyclo[2.2.1]heptane-3-carboxylic acid; 2-
azabicyclo[3.1.0]hexane-3-carboxylic acid; 2-azaspiro[4.4]nonane-3-carboxylic
acid;
2-azaspiro[4.5]decane-3-carboxylic acid; spiro(bicyclo[2.2.1)heptane)-2,3-
pyrrolidine-5-carboxylic acid; spiro(bicyclo[2.2.2]octane)-2,3-pyrrolidine-5-
carboxylic
acid; 2-azatricyclo[4.3Ø16~9]decane-3-carboxylic acid;
decahydrocyclohepta[b]pyrrole-2-carboxylic acid; decahydrocycloocta[c]pyrrole-
2-
carboxylic acid; octahydrocyclopenta[c]pyrrole-2-carboxylic acid;
octahydroisoindole-1-carboxylic acid; 2,3,3a,4,6a-
hexahydrocyclopenta[b]pyrrole-2-
carboxylic acid; 2,3,3a,4,5,7a-hexahydroindole-2-carboxylic acid;
tetrahydrothiazole-
4-carboxylic acid; isoxazolidine-3-carboxylic acid; pyrazolidine-3-carboxylic
acid,
hydroxypyrrolidine-2-carboxylic acid; all of which can be optionally
substituted (see
following formulae):
. * CO- . CO- .
* i
N CO_ ~ * , I
CO- ~ N~ N~
~ ~ ~CO- .
N_ -kC0- ~ v 'N' '*CO- ~ " _N
CA 02225366 1997-12-19
CO- ; CO- ; ~CO- ;
N
* CO- ~ * CO- ~ CO- ;
~N\
CO- ; CO- . CO- ;
~N
* ~ >---CO- .
CO-; * ; * ; ,
N~-'CO- N J--CO- N
* S
CO- ; ~--CO- ; O~-CO- ; N~-CO- ;
Nr ~ N N N
CA 02225366 1997-12-19
16
HO
~-CO-
N
The heterocycles on which the abovementioned radicals are based are disclosed,
for example, in US-A-4,344,949; US-A 4,374,847; US-A 4,350,704; EP-A 29,488;
E P-A 31, 741; E P-A 46, 953; E P-A 49, 605; E P-A 49, 658; E P-A 50, 800; E P-
A 51, 020;
E P-A 52, 870; E P-A 79, 022; E P-A 84,164; E P-A 89, 637; E P-A 90, 341; E P-
A 90, 362;
EP-A 105,102; EP-A 109,020; EP-A 111,873; EP-A 271,865 and EP-A 344,682.
Furthermore, the amino acids can also be present as esters or amides, such as,
for
example, as the methyl ester, ethyl ester, isopropyl ester, isobutyl ester,
tert-butyl
ester, benzyl ester, unsubstituted amide, methylamide, ethylamide,
semicarbazide
or c~-amino-(C2-C8)-alkylamide.
Functional groups of the amino acids can be protected. Suitable protective
groups
such as, for example, urethane protective groups, carboxyl protective groups
and
side-chain protective groups are described in Hubbuch, Kontakte (Merck) 1979,
No.
3, pages 14 to 23 and in Bullesbach, Kontakte (Merck) 1980, No. 1, pages 23 to
35.
The following may be mentioned in particular: Aloc, Pyoc, Fmoc, Tcboc, Z, Boc,
Ddz, Bpoc, Adoc, Msc, Moc, Z(N02), Z(Hah), Bobz, Iboc, Adpoc, Mboc, Acm, tert-
Butyl, OBzI, ONbzl, OMbzl, Bzl, Mob, Pic, Trt.
The compounds of the formulae I and la according to the invention may be
present
as E/Z isomers. The invention relates to pure E isomers and pure Z isomers as
well
as to EIZ isomer mixtures in all ratios. The compounds of the formulae I and
la can
contain optically active carbon atoms which independently of one another may
have
the R- or S-configuration. They can be present in the form of pure enantiomers
or
pure diastereomers or in the form of enantiomer mixtures, for example in the
form of
racemates, or diastereomer mixtures. The invention relates to both pure
enantiomers and enantiomer mixtures in all ratios and diastereomers and
CA 02225366 1997-12-19
17
diastereomer mixtures in all ratios. Diastereomers, including EIZ isomers, can
be
separated into the individual isomers, for example, by chromatography.
Racemates
can be separated into the two enantiomers, for example, by chromatography on
chiral phases or by resolution. If mobile hydrogen atoms are present, the
present
invention also includes all tautomeric forms of the compounds of the formulae
I and
la.
Physiologically tolerable salts of the compounds of the formulae I and la are,
in
particular, pharmaceutically utilizable or nontoxic, physiologically
utilizable salts.
Such salts of compounds of the formulae I and la which contain acidic groups,
for
example carboxyl, are, for example, alkali metal salts or alkaline earth metal
salts,
such as, for example, sodium salts, potassium salts, magnesium salts and
calcium
salts, and also salts with physiologically tolerable quaternary ammonium ions
and
acid addition salts with ammonia and physiologically tolerable organic amines,
such
as, for example, triethylamine, ethanolamine or tris(2-hydroxyethyl)amine.
Compounds of the formulae I and la, which contain basic groups, for example
one or
more amino groups, amidino groups or guanidino groups, form acid addition
salts,
for example with inorganic acids such as hydrochloric acid, sulfuric acid or
phosphoric acid, or with organic carboxylic acids and sulfonic acids such as
acetic
acid, citric acid, benzoic acid, malefic acid, fumaric acid, tartaric acid,
methanesulfonic acid or p-toluenesulfonic acid.
A physiologically tolerable anion Q-, which is contained in the compounds of
the
formulae 1 and la when R8 and/or R8~ is the 2-trimethylammonio-ethoxy radical,
is, in
particular, a monovalent anion or an equivalent of a polyvalent anion of a
nontoxic,
physiologically utilizable, in particular also pharmaceutically utilizable,
inorganic or
organic acid, for example the anion or an anion equivalent of one of the
abovementioned acids suitable for the formation of acid addition salts. Q- can
thus
be, for example, one of the anions (or an anion equivalent) from the group
consisting of chloride, sulfate, phosphate, acetate, citrate, benzoate,
maleate,
fumarate, tartrate, methanesulfonate and p-toluenesulfonate.
CA 02225366 1997-12-19
18
Salts can be obtained from the compounds of the formulae I and la by customary
methods known to those skilled in the art, for example by combining the
compounds
of the formulae I and la with an inorganic or organic acid or base in a
solvent or
dispersant, or also from other salts by ration exchange or anion exchange. The
present invention also includes all salts of the compounds of the formulae I
and la
which, because of low physiological tolerability, are not directly suitable
for use in
pharmaceuticals, but are suitable, for example, as intermediates for carrying
out
other chemical modifications of the compounds of the formulae I and la or as
starting materials for the preparation of physiologically tolerable salts.
The present invention moreover includes all solvates of compounds of the
formulae
I and la, for example hydrates or adducts with alcohols, and also derivatives
of the
compounds of the formulae I and la, for example esters, prodrugs and
metabolites,
which act like the compounds of the formulae I and la. The invention relates
in
particular to prodrugs of the compounds of the formulae I and la, which can be
converted into compounds of the formulae I and la under physiological
conditions.
Suitable prodrugs for the compounds of the formulae I and la, i.e. chemically
modified derivatives of the compounds of the formulae I and la having
properties
which are improved in a desired manner, are known to those skilled in the art.
More
detailed information relating to prodrugs is found, for example, in Fleisher
et al.,
Advanced Drug Delivery Reviews 19 (1996) 115-130; Design of Prodrugs, H.
Bundgaard, Ed., Elsevier, 1985; H. Bundgaard, Drugs of the Future 16 (1991 )
443;
Saulnier et al., Bioorg. Med. Chem. Lett. 4 (1994) 1985; Safadi et al.,
Pharmaceutical Res. 10 (1993) 1350. Suitable prodrugs for the compounds of the
formulae I and la are especially ester prodrugs of acid groups, for example of
carboxylic acid groups, in particular of a COOH group representing R4 , and
also
aryl prodrugs and carbamate prodrugs of acylatable nitrogen-containing groups
such as amino groups, amidino groups or guanidino groups, in particular of the
groups Rs-C(=NRs)-NRs-, RsR6~N-C(=NRs)-, RsR6~N-C(=NR6)-NR6 - and the 4- to
11-membered, monocyclic or polycyclic, aromatic or nonaromatic ring system
representing the group E. In the aryl prodrugs or carbamate prodrugs, a
hydrogen
atom located on a nitrogen atom is replaced one or more times, for example
twice,
CA 02225366 1997-12-19
19
in these groups by an acyl group or carbamate group. Suitable acyl groups and
carbamate groups for the acyl prodrugs and carbamate prodrugs are, for
example,
the groups R6-CO and R60-CO, in which R6 has the meanings indicated above,
i.e.
hydrogen, (C~-C~8)-alkyl, (C3-C~4)-cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-
alkyl, (C5-
C~4)-aryl, in which 1 to 5 carbon atoms can be replaced by heteroatoms such as
N,
O, S, or (C5-C~4)-aryl-(C~-C$)-alkyl, in which 1 to 5 carbon atoms in the aryl
moiety
can be replaced by heteroatoms such as N, O, S, combinations of substituent
meanings which in the individual case .lead to unstable compounds, for example
to
unstable free carbamic acids, not being suitable. These prodrugs can be
prepared
by customary methods familiar to those skilled in the art for the preparation
of
acylamines and carbamates.
The present invention is furthermore not restricted to the compounds according
to
the formulae I and la having a purine parent structure, but also includes
those
compounds which instead of the purine parent structure shown in the formulae I
and
la have a 3-deazapurine structure, 7-deazapurine structure or 7-deaza-8-
azapurine
structure, i.e. compounds of the formulae Ib and Ic, Id and 1e and If and Ig.
Wa
N N N
\ ~~Y N \ ~~Y
/ N / N
X
X G (1b) Ga (lc)
W W a
N \ ~ Y N \ ~ Y
Nr
X ~ X N ~a
G (Id) G (1e)
CA 02225366 1997-12-19
Wa
N \ ~N N \ ~ N
N N N N.
,
5 X ~ X I
G (I~ Ga (/g)
All the above and following details relating to the compounds of the formulae
I and
la apply to the compounds of the formulae Ib and Ic, Id and 1e, If and Ig
10 correspondingly. If compounds of the formulae I and la are being discussed,
then, if
not stated otherwise, the deaza analogs and deaza-aza analogs of the formulae
Ib
and Ic, Id and 1e, If and Ig are also included. Preferably, in the compounds
according to the invention the purine structure actually shown in formulae I
and la is
present, in which the nitrogen atoms are present in the 3-position and in the
7-
15 position and a carbon atom with the group Y bonded thereto is present in
the 8-
position.
In the compounds of the formulae I and la, X is preferably hydrogen, NR6R6 ,
hydroxy-(C~-Cs)-alkyl or NH-CO-Rs, particularly preferably hydrogen, NR6R6~ or
20 NH-CO-R6, very particularly preferably hydrogen or NH2. Y is preferably
hydrogen.
R4 is preferably C(O)R8. Preferred compounds according to the invention are
also
those of the formulae I and la in which R3 is RsRs~N-R~, RsOC(O)N(R5)R~,
RsS(O)pN(R5)R7, RsC(O)N(R5)R~ or R6N(Rs~)C(0)N(R5)R~, where p here is 1 or 2,
in particular compounds in which R3 is RsOC(O)N(R5)R7 or RsS(O)PN(RS)R~ (where
p = 1 or 2); particularly preferred compounds here are those in which a
lipophilic
radical is contained in R3, for example compounds in which R6 and/or R6~, for
example in the group RsOC(O)N(R5)R7, is (C4-C~4)-alkyl, (CS-C~4)-aryl-(C~-C4)-
alkyl, for example benzyl, (CS-C~4)-cycloalkyl or (C5-C~4)-cycloalkyl-(C~-C4)-
alkyl,
preferred cycloalkyl radicals here in particular being the 1-adamantyl radical
and the
2-adamantyl radical.
A preferred group of compounds according to the invention is formed by
compounds
of the formulae I and la in which:
CA 02225366 1997-12-19
21
X is hydrogen, NH2, OH or NH-CO-R6;
Y is hydrogen;
G is a radical of the formula II
-(CR~R2)~-A-(CR~R2)m-(CR~R3)i (CR~R2)q-R4 (II);
W is a radical of the formula III
-B-(CRS R2)~ A'-(CRS R2)$ (CRS R3)k-(CRS R2)t-D-E (III);
Ga is a radical of the formula Ila
-(CRS R2)~ A'-(CRS RZ)g (CRS R3)k-(CRS R2)t-D-E (11a);
Wa is a radical of the formula Illa
-B-(CRS R2)~ A-(CRS R2)m-(CRS R3)i (CRS R2)q R4 (II la);
A, A' independently of one another are a direct bond, -C(O)NRS-, -NR5C(O)-,
-C(O)-, -NR5-, -0- , -S-, -SO-, -S02-, (C5-C~4)-arylene, it being possible in
the aryl radical for one to five carbon atoms to be replaced by one to five
heteroatoms, (C2-C4)-alkynylene, (C2-C4)-alkenylene, or a divalent radical of
a 3- to 7-membered saturated or unsaturated ring, which can contain one or
two heteroatoms, such as, for example, nitrogen, sulfur or oxygen and which
can be monosubstituted or disubstituted by =O, =S or R3 ;
R~, R2 independently of one another are H, fluorine, chlorine, CN, nitro, (C~-
C~o)-
alkyl, (C3-C~4)-cycloalkyl, (C3-C~2)-cycloalkyl-(C~-C$)-alkyl, (C5-C~4)-aryl,
(C5-C~4)-aryl-(C~-C8)-alkyl, Rs-O-R ~, Rs-S(O)p-R~ or RsRs~N-R~;
R3 independently of one another is H, fluorine, chlorine, CN, nitro, (C~-C~4)-
CA 02225366 1997-12-19
22
alkyl, (C3-C~4)-cycloalkyl, (C3-C~4)-cycloalkyl-{C~-C8)-alkyl, (C5-C~4)-aryl,
{C5-C~4)-aryl-(C~-C8)-alkyl, Rs-O-R~, R6-S(O)S R7, R6Rs~N-R7, RsCO2R~,
RsCOR~, RsOC(O)R~, R6N{R5)C(O)OR~, R6S(O)PN(R5)R~,
RsOC(O)N(R5)R7, RsC(O)N(RS)R~, RsN(R5)C(O)N(R5)R7,
RsN(RS)S{O)pN(R5)R~, RsS(O)pR~, RsSC(O)N(R5)R~, R6C(O)Rs,
R6N(R5)C(O)R~ or R6N(R5)S(0)pR~, it being possible for alkyl to be
monounsaturated or polyunsaturated and it furthermore being possible for
alkyl and aryl to be monosubstituted or polysubstituted by fluorine, chlorine,
bromine, CN, RsN(R5)R7, RsR6~NR~, nitro, R60C(O)R~, R6C(O)R~,
RsN(R5)C(O)R~, RsN(R5)S(O)pR~, Rs, R6-O-R7;
R4 is C(O)R8, C(S)R8, S{O)PR8, POR8R8~, an L- or D-amino acid or a four- to
eight-membered, saturated or unsaturated heterocycle which contains 1, 2, 3
or 4 heteroatoms from the group consisting of N, O, S, such as, for example,
tetrazolyl, imidazolyl, pyrazolyl, oxazolyl, thiadiazolyl;
R5 is H, {C~-Coo)-alkyl, (C3-C~4)-cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-
alkyl,
(C5-C~4)-aryl or (C~-C~4)-aryl-(C~-C8)-alkyl;
Rs, Rs~ independently of one another are H, (C~-C8)-alkyl, (C3-C~4)-
cycloalkyl, (C3-
C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, it being possible for 1 - 5
carbon
atoms to be replaced by heteroatoms, or (C5-C~4)-aryl-(C~-C8)-alkyl, it being
possible for 1 - 5 carbon atoms in the aryl moiety to be replaced by
heteroatoms, or R6 and Rs~, together with the atoms connecting them, form a
ring system which can optionally also contain further heteroatoms from the
group consisting of N, S, O, such as, for example, morpholine, piperazine,
piperidine, pyrrolidine;
R~ independently of one another is (C~-C4)-alkylene or a direct bond;
R8, R8~ independently of one another are OH, (C~-C8)-alkoxy, (C5-C~4)-aryl-(C~-
C$)-
alkoxy, (C5-C~4)-aryloxy, (C~-C8)-alkylcarbonyloxy-(C~-C4)-alkoxy, (C5-C~4)-
aryl-(C~-C8)-alkylcarbonyloxy(C~-Cs)-alkoxy, NR6R6~, (C~-C8)-
CA 02225366 1997-12-19
23
dialkylaminocarbonylmethyloxy, (C5-C~4)-aryl-(C~-C8)-
dialkylaminocarbonylmethyloxy, (C5-C~4)-arylamino or an L- or D-amino
acid;
B is O, S, NRS, -NRS-C(O)-, -C(O)-NR5-, a direct bond or a divalent radical of
a
3- to 7-membered saturated or unsaturated ring which can contain one or
two heteroatoms, such as, for example, nitrogen, sulfur or oxygen, and which
can be monosubstituted or disubstituted by =O, =S or R3;
D is a direct bond, -NRs-, -C(O)-NRs-, -NRs-C(O)- , -S02NR6-,
-NRs-C(O)-NR6-, -NR6-C(S)-NRs-, -NR6-S(O)S NRs-, -NRs-C(O)O-,
-NRs-N=CRs-, -NRs-S(O)~ , -(CS-C~4)-aryl-CO-, -(CS-C~4)-aryl-S(O)S ,
-N=CRs-, -RsC=N- or -RsC=N-NR6- ;
E is hydrogen, Rs-C(=NR6)NRs-, RsR6~N-C(=NRs)-, RsRs~N-C(=NRs)-NRs-, or
a 4- to 11-membered, mono- or polycyclic, aromatic or nonaromatic ring
system which can optionally contain 1- 4 heteroatoms from the group
consisting of N, O and S and can optionally be monosubstituted to
trisubstituted by R3, R5, =O, =S or RsRs~N-C(=NRs)-, such as, for example,
the radicals indicated with their structural formulae in the above definition
of
E;
n is zero, one, two, three, four or five;
m is zero, one, two, three, four or five;
i is zero or one;
p independently of one another is zero, one or two;
q independently of one another is zero, one or two;
r is zero, one, two, three, four, five or six;
s is zero, one, two, three, four or five;
t is zero, one, two, three, four or five;
k is zero or one;
a is one or two;
v in the radicals indicated in the above definition of E is the numbers zero,
one, two
CA 02225366 1997-12-19
24
or three;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts, where, in this group of preferred compounds,
the
analogs having a 3-deazapurine structure, 7-deazapurine structure or 7-deaza-8-
azapurine structure are not included.
A further group of preferred compounds is formed by compounds of the formulae
I
and la in which:
X is hydrogen, NRsRs~, hydroxy-(C~-Cs)-alkyl-NH or NH-CO-Rs;
Y is hydrogen;
G is a radical of the formula II
-(CRS R2)~ A-(CRS R2)m-(CRS R3)i-(CRS R2)q-R4 (II);
W is a radical of the formula III
-B-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-D-E (III);
Ga is a radical of the formula Ila
-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-D-E (I la);
Wa is a radical of the formula Illa
-B-(CRS R2)~-A-(CRS R2)m-(CRS R3)i-(CRS R2)q R4 (II la);
A, A' independently of one another are a direct bond, -C(O)NR5-, -NRSC(O)-,
-C(0)-, -NR5-, -O-, -S-, -SO-, -S02-, (C5-C~4)-arylene, it being possible
for one to three carbon atoms in the aryl radical to be replaced by one to
CA 02225366 1997-12-19
three heteroatoms from the group consisting of O, N, S, (C2-C4)-alkynylene or
(C2-C4)-alkenylene;
R~, R2 independently of one another are hydrogen, fluorine, cyano, (C~-C4)-
alkyl,
5 (C5-C6)-aryl, (C5-C6)-aryl-(C~-C4)-alkyl, R6-O-R~ or R6R6~N-R~;
R3 independently of one another is hydrogen, (C~-C~8)-alkyl, (C3-C~4)-
cycloalkyl,
(C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, (C5-C~4)-aryl-(C~-C8)-alkyl,
R6R6~N-R~, R6C(O)R~, R6S(O)PN(R5)R~, RsOC(O)N(R5)R~, RsC(O)N(R5)R7,
10 R6N(R6~)C(O)N(R5)R~, R6N(Rs~)S(O)pN(R5)R~ or RsN(Rs~)C(O)R~, it being
possible for alkyl to be monounsaturated or polyunsaturated and it
furthermore being possible for alkyl and aryl to be monosubstituted or
polysubstituted by fluorine, chlorine, bromine, cyano, RsRs NR~, nitro,
R60C(O)R~, RsC(O)R~, R6N(Rg~)C(O)R~, R6N(Rs~)S(O)pR~, Rs or RsORl;
R4 is C(O)R8;
RS independently of one another is hydrogen or (C~-C4)-alkyl;
Rs, Rs~ independently of one another are hydrogen, (C~-C~8)-alkyl, (C3-C~4)-
cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, in which one to
three carbon atoms can be replaced by one to three heteroatoms from the
group consisting of N, S, O, or are (CS-C~4)-aryl-(C~-C8)-alkyl, in which one
to three carbon atoms in the aryl moiety can be replaced by one to three
heteroatoms from the group consisting of N, S, O, or Rs and Rs~, together
with the atoms connecting them, form a ring system which can optionally
also contain additional heteroatoms from the group consisting of N, S, O,
such as, for example, morpholine, piperazine, piperidine, pyrrolidine;
R~ independently of one another is (C~-C2)-alkylene or a direct bond;
R8 independently of one another is hydroxyl, (C~-C4)-alkoxy, (C5-C~4)-aryl-(C~-
C4)-alkoxy, (C5-C~4)-aryloxy, (C~-C8)-alkylcarbonyloxy-(C~-C4)-alkoxy, (C5-
CA 02225366 1997-12-19
26
C~4)-aryl-(C~-C4)-alkylcarbonyloxy-(C~-C4)-alkoxy or the radical of an amino
acid;
B is -O-, -S-, -NR5-, a direct bond or a divalent radical of a 3- to 7-
membered
saturated or unsaturated ring which can contain one or two heteroatoms,
such as, for example, nitrogen, sulfur or oxygen and which can be
monosubstituted or disubstituted by radicals from the group consisting of
=O, =S and R3;
D is a direct bond, -NRs-, -C(O)-NR6-, -NR6-C(O)- , -NRs-C(O)-NRs-,
-NRs-C(O)O-, -NRs-N=CR6-, -RsC=N-NR6-, -N=CRs- or -RsC=N-, where
the divalent radicals representing D are bonded to the group E via the free
bond on the right side;
E is hydrogen, R6-C(=NRs)-NRg~-, RsR6~N-C(=NR6~)-, R6R6~N-C(=NRs~)-NRs-
or a radical from the group consisting of
~s Rs R5 15
R3 N R3 N N
~ / R3 v
N ,
N N N '
O
H N
N Ra ~ N R3 / I \~--
\~ \ I \~ ~N N
N ,
' , ~R5
H Rs R
CA 02225366 1997-12-19
27
H R5
N N N\
N\ ~ N NH ,
H Rs
O
which can optionally be monosubstituted to trisubstituted by radicals from
the group consisting of R3, R5, =O, =S and RsRs~N-C(=NRs)-;
n is one, two, three or four;
m is zero or one;
i is zero or one;
q is zero or one;
p independently of one another is zero,
one or two;
r is zero, one, two, three, four or five;
s is zero, one or two;
t is zero, one or two;
k is zero or one;
v is zero, one, two or three;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts and their prodrugs.
Particularly preferred compounds of the formulae I and la are those in which:
X is hydrogen, NRsRs~ or NH-CO-Rs;
Y is hydrogen;
G is a radical of the formula II
-(CRS R2)~-A-(CRS R2)m-(CRS R3)i-(CRS R2)q-R4 (II);
CA 02225366 1997-12-19
28
W is a radical of the formula III
-B-(CR~R2)~ A'-(CR~R2)S (CR~R3)k-(CR~RZ)t-D-E (III);
Ga is a radical of the formula Ila
-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-D-E (11a);
Wa is a radical of the formula Illa
-B-(CRS R2)~ A-(CRS R2)m-(CRS R3)i (CRS R2)q-R4 (II la);
A, A' independently of one another are a direct bond, -C(O)NR~-, -NRSC(O)- or
(C5-Cs)-arylene, it being possible for one to two carbon atoms in the aryl
radical to be replaced by nitrogen atoms;
R~ , R2 are hydrogen;
R3 independently of one another is hydrogen, (C~-C~o)-alkyl, (C3-C~4)-
cycloalkyl, (C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (CS-C~4)-aryl, (C5-C~4)-aryl-
(C~-C8)-alkyl, RsR6~N-R~, RsOC(O)N(R5)R7, RsC(O)N(R5)R~,
RsN(Rs~)C(O)N(R5)R~, R6C(O)R7 or RsN(R6~)C(O)R~, it being possible for
alkyl to be monounsaturated or polyunsaturated and it furthermore being
possible for alkyl and aryl to be monosubstituted or polysubstituted by
fluorine, chlorine, bromine, cyano, R6R6~NR~, R6C(O)R7, R6N(Rs~)C(O)R~,
Rs or RsOR~;
R4 is C(O)R8;
R5 independently of one another is hydrogen or (C~-C4)-alkyl;
Rs, R6~ independently of one another are hydrogen, (C~-C$)-alkyl, (C3-C~2)-
cycloalkyl, (C3-C~2)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, in which one to
CA 02225366 1997-12-19
29
three carbon atoms can be replaced by one to three heteroatoms from the
group consisting of N, S, O, or are (C5-C~4)-aryl-(C~-C8)-alkyl, in which one
to three carbon atoms in the aryl moiety can be replaced by one to three
heteroatoms from the group consisting of N, S, O;
R7 is a direct bond;
R8 independently of one another is hydroxyl, (C~-C4)-alkoxy, (C5-C~4)-aryl-(C~-
C4)-alkoxy, (CS-C~4)-aryloxy, (C~-C8)-alkylcarbonyloxy-(C~-C4)-alkoxy, (C5-
C~4)-aryl-(C~-C4)-alkylcarbonyloxy-(C~-C4)-alkoxy or the radical of an amino
acid;
B is -O-, -S-, -NRS-, a direct bond or a divalent radical of a 3- to 7-
membered
saturated or unsaturated ring, which can contain one or two heteroatoms,
such as, for example, nitrogen, sulfur or oxygen and which can be
monosubstituted or disubstituted by radicals from the group consisting of =O,
=S and R3 ;
D is a direct bond, -NRs-, -C(O)-NRs- or -NR6-C(O)-;
E is hydrogen, Rs-C(=NR6)-NR6~-, RsRs~N-C(=NRs~)-, R6Rs~N-C(=NR6~)-NRs-
or a radical from the group consisting of
Rs Rs R5
R3 N R3 N N
/~ R3
N ~ N N
N , N
R3 ~ ~ R3 ~ ~>--
N ~N N
1R5 ~R5
CA 02225366 1997-12-19
which can optionally be monosubstituted to trisubstituted by radicals from the
group consisting of R3, R5, =O, =S and RsR6~N-C(=NRs)-;
r is zero, one, two, three, four or five;
5 s is zero or one;
t is zero or one;
k is zero or one;
n is one, two, three or four;
m is zero or one;
10 i is zero or one;
q is zero or one;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts and their prodrugs.
Particularly preferred compounds of the formula I are furthermore those in
which:
X is hydrogen, NR6R6~ or NH-CO-R6;
Y is hydrogen;
G is a radical of the formula II
-(CRS R2)~-A-(CRS R2)m-(CRS R3)~ (CRS R2)q-R4 (II);
W is a radical of the formula III
-B-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS RZ)t-D-E (III);
A, A' are a direct bond;
R~, R2 independently of one another are hydrogen, (C~-C4)-alkyl, (C5-Cs)-aryl
or (C5-Cs)-aryl-(C~-C4)-alkyl;
CA 02225366 1997-12-19
31
R3 independently of one another is hydrogen, (C~-C~8)-alkyl, (C3-C~4)-
cycloalkyl,
(C3-C~4)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, (C5-C~4)-aryl-(C~-C8)-alkyl,
RsRs~N-R7, R60C(O)N(R5)R7, RsS02N(R5)R~, RsC(O)N(R5)R7,
R6N(R6~)C(O)N(R5)R~, R6C(O)R7 or RsN(R6~)C(O)R~, it being possible for
alkyl to be monounsaturated or polyunsaturated and it furthermore being
possible for alkyl and aryl to be monosubstituted or polysubstituted by
fluorine, chlorine, bromine, cyano, RsR6~NR~ , RsC(O)R~, R6N(R6~)C(O)R7,
R6 or RsOR~;
R4 is C(O)R8;
RS independently of one another is hydrogen or (C~-C4)-alkyl;
Rs, R6~ independently of one another are hydrogen, (C~-C~8)-alkyl, (C3-C~2)-
cycloalkyl, (C3-C~2)-cycloalkyl-(C~-C8)-alkyl, (CS-C~4)-aryl, in which 1 to 3
carbon atoms can be replaced by 1 to 3 heteroatoms from the group
consisting of N, S, 0, or are (C5-C~4)-aryl-(C~-C8)-alkyl, in which 1 to 3
carbon atoms in the aryl radicals can be replaced by 1 to 3 heteroatoms
from the group consisting of N, S, O, and it also being possible for Rs and
R6~, together with the atoms connecting them, to form a ring system which
can optionally also contain additional, in particular one, two or three,
heteroatoms from the group consisting of N, S, O;
R~ is a direct bond;
R8 independently of one another is hydroxyl, (C~-C4)-alkoxy, (C5-C~4)-aryl-(C~-
C4)-alkoxy, (CS-C~4)-aryloxy, (C~-C8)-alkylcarbonyloxy(C~-C4)-alkoxy or (C5-
C~4)-aryl-(C~-C4)-alkylcarbonyloxy(C~-C4)-alkoxy;
B is 1,4-piperidinediyl or 1,4-piperazinediyl, where in the case of the 1,4-
piperidinediyl radical the nitrogen atom of the piperidine is bonded to the
purine structure;
CA 02225366 1997-12-19
32
D is a direct bond, -NRs-, -C(O)-NRs- or -NRs-C(O)-;
E is hydrogen, Rs-C(=NR6)NR6~-, R6R6~N-C(=NRs~)-, R6R6'N-C-(=NR6')-NRs-
or a radical from the group consisting of
Rs Rs R5
Rs N R3 N 3 N
/ /~ R
N '
N , N ,
N , N
R3 ~ \ R3
N ~N N ,
5
1R5 R
which can optionally be monosubstituted to trisubstituted by radicals from the
group consisting of R3, R5, =0, =S and R6R6'N-C(=NRs)-;
r is zero, one
or two;
s is zero or one;
t is zero or one;
k is zero or one;
n is zero, one
or two;
m is zero or one;
i is zero or one;
q is zero or one;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts and their prodrugs.
Very particularly preferred compounds of the formula I are those in which:
CA 02225366 1997-12-19
33
X is hydrogen;
Y is hydrogen;
G is a radical of the formula II
-(CRS R2)~-A-(CRS R2)m-(CRS R3)i-(CRS RZ)q-R4 (I I).
W is a radical of the formula III
-B-(CRS R2)~ A'-(CRS R2)g (CRS R3)k-(CRS R2)t-D-E (I II);
A, A' are a direct bond;
R~, R2 independently of one another are hydrogen or (C~-C2)-alkyl, in
particular
hydrogen;
R3 is RsR6~N-R7, RsOC(O)N(R5)R~, R6S02N(R5)R7, RsC(O)N(R5)R~ or
RsN(Rs~)C(O)N(R5)R7, in particular RsOC(O)N(RS)R7 ;
R4 is C(O)R8;
R5 is hydrogen or (C~-C2)-alkyl, in particular hydrogen;
R6, Rs~ independently of one another are hydrogen, (C~-C~8)-alkyl, (C3-C~2)-
cycloalkyl, (C3-C~2)-cycloalkyl-(C~-C8)-alkyl, (C5-C~4)-aryl, in which 1 to 3
carbon atoms can be replaced by 1 to 3 heteroatoms from the group
consisting of N, S, 0, or are (C5-C~4)-aryl-(C~-C8)-alkyl, in which 1 to 3
carbon atoms in aryl radicals can be replaced by 1 to 3 heteroatoms from
the group consisting of N, S, O, and it also being possible for Rs and Rs~,
together with the atoms connecting them, to form a ring system which can
optionally also contain additional, in particular one, two or three,
heteroatoms from the group consisting of N, S, 0;
CA 02225366 1997-12-19
34
R7 is a direct bond;
R8 is hydroxyl, (C~-C4)-alkoxy, (C5-C~4)-aryl-(C~-C4)-alkoxy, (C5-C~4)-
aryloxy,
(C~-C8)-alkylcarbonyloxy(C~-C4)-alkoxy or (C5-C~4)-aryl-(C~-C4)-
atkylcarbonyloxy-(C~-C4)-alkoxy, in particular hydroxyl or (C~-C4)-alkoxy;
B is 1,4-piperidinediyl, where the nitrogen atom of the piperidine is bonded
to
the purine structure;
D is -NR6- or -C(O)-NRs-, where in the group -C(O)-NR6- the nitrogen atom is
bonded to the group E;
E is R6R6~N-C(=NRs~)- or a radical from the group consisting of
Rs Rs R5
Ra N Ra N 3 N
/ / R
N N , N ,
R3 ~ ( N~ R3 / ~ N
W N wN~N
\ 5
1R5 R
which can optionally be monosubstituted to trisubstituted by radicals from the
group consisting of R3, R5, =O, =S and RsR6~N-C(=NRs)-;
r is zero or one;
s is zero;
t is zero;
CA 02225366 1997-12-19
k is zero;
n is one;
m is zero;
is one;
5 q is zero;
in all their stereoisomeric forms and mixtures thereof in all ratios, and
their
physiologically tolerable salts and their prodrugs.
10 Especially preferred compounds according to the invention are the compounds
of
the formula Ih
H H
H N H N
O N ~\ O N ~\
15 N N
N N
(1h) (1k)
N N
20 NI ~ ~~ NI
~N N ~N N
R" S COOH
(h) 3
R / O NH
25 0
in which R3 is RsRs~N-R7, RsOC(0)N(R5)R~, R6S02N(R5)R~, RsC(0)N(R5)R7 or
R6N(R6~)C(O)N(R5)R~, in particular RsOC(O)N(R5)R7, and Rh is the carboxylic
acid
group COOH or a carboxylic acid derivative, for example an ester such as, for
30 example, a (C~-C4)-alkyl ester, i.e. for example the group COO-(C~-C4)-
alkyl; in all
their stereoisomeric forms and mixtures thereof in all ratios, and their
physiologically
tolerable salts and their prodrugs. In compounds of the formula Ih in which R~
is a
direct bond, the stereochemical center (h) in the formula Ih preferably has
the S-
CA 02225366 1997-12-19
36
configuration. Compounds of the formula Ih in which R7 is a direct bond can be
named as 2-amino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-ylcarbamoyl)piperidin-
1-
yl)purin-9-yl)propionic acid and derivatives thereof, for example esters,
optionally
substituted on the 2-amino group. A particularly especially preferred compound
is
2S-benzyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-yl-carbamoyl)-
piperidin-1-yl)-purin-9-yl)-propionic acid of the formula Ik and its
physiologically
tolerable salts and its prodrugs.
Compounds of the formulae I and la can generally be prepared, for example in
the
course of a convergent synthesis, by linkage of two or more fragments which
can be
derived retrosynthetically from the formulae I and la. In the preparation of
the
compounds of the formulae I and la, it can generally be advantageous or
necessary
in the course of the synthesis to introduce functional groups which could lead
to
undesired reactions or side reactions in the respective synthesis step, in the
form of
precursors which are later converted into the desired functional groups, or
temporarily to block functional groups by a protective group strategy suited
to the
synthesis problem, what is known to those skilled in the art (Greene, Wuts,
Protective Groups in Organic Synthesis,Wiley, 1991 ).
The present invention also relates to processes for the synthesis of the
compounds
of the formula I, which comprise carrying out one or more of the following
steps for
the synthesis of the compounds of the formula 1.
a1 ) A compound of the formula IV,
1
N
yY (IV)
N~ N
in which
L1 is a customary leaving group known to those skilled in the art, for example
chlorine, bromine, iodine, OTos or OMes, preferably chlorine or bromine, and
CA 02225366 1997-12-19
37
X and Y are as defined above, but functional groups can optionally also be
present
in the form of precursors or can temporarily be protected by a protective
group,
is reacted with a compound of the formula V
L2-(CRS R2)~-A-(CRS R2)m-(CRS R3)~ (CRS R2)q R~ ~ (V)
in which
R~, R2, R3, A, n, m, i and q are as defined above,
R~~ is defined as R4 above, but is optionally protected by a protective group,
for
example for R4 = COOH by a tert-butyl or a methyl or ethyl protective group,
L2 is hydroxyl or a leaving group known to those skilled in the art, for
example
chlorine, bromine, iodine, OTos, OMes or OTf,
to give a compound of the formula VI
1
N
N ~ yY NI)
~N N
R"
in which
R~ ~ is -(CRS R2)~ A-(CRS R2)m-(CRS R3)~-(CRS R2)q R~ ~ and for which
otherwise the
above meanings apply,
the reaction being carried out according to methods known to those skilled in
the art
(see source literature in J. March, Advanced Organic Chemistry, Fourth
Edition,
Wiley, 1992). Preferably, the reaction is carried out in a suitable organic
solvent or
diluent, for example DCM, CHC13, THF, diethyl ether, n-heptane, n-hexane, n-
CA 02225366 1997-12-19
38
pentane, cyclohexane, diisopropyl ether, methyl tert-butyl ether,
acetonitrile, DMF,
DMSO, dioxane, toluene, benzene, ethyl acetate or a mixture of these solvents,
if
appropriate with addition of a base such as, for example, butyllithium,
lithium
diisopropylamide (LDA), sodium hydride, sodium amide, potassium tart-butoxide,
CaC03, Cs2C03, triethylamine, diisopropylethylamine or complex bases (sodium
amidelR~20Na, where R~2 is (C2-C6)-alkyl or CH3CH20CH2CH2). For L2 = OH, the
reaction can be carried out, for example, by the conditions described for the
Mitsunobu reaction (Hughes, Organic Reactions 42 (1992) 335-656), for example
by
reaction with triphenylphosphine and DEAD in THF.
a2) The compound of the formula VI is reacted with a compound of the formula
VII
H-B-(CRS R2)~ A'-(CRS RZ)S (CRS R3)k-(CRS R2)t-R~ 3 (VI I)
in which R~3 is -D-E or a group R~4 which can be converted into D-E and which
is
optionally provided with suitable protective groups, and for which otherwise
the
above meanings apply. R~4 is, for example, an optionally protected amino group
-NHRs, it being possible to employ, for example, the Boc protective group as a
protective group, a protected carboxylic acid ester, an aldehyde -C(O)H, a
keto
group -C(O)RE, or a protected mercapto group.
In this reaction, a compound of the formula VIII
R~s
N ~ N
(VIII)
N~ N
R"
is obtained, in which
R~5 is -B-(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-R~3 and for which
otherwise
the above meanings apply.
CA 02225366 1997-12-19
39
The reaction is carried out according to methods known to those skilled in the
art
(see source literature in J. March, Advanced Organic Chemistry, Fourth
Edition,
Wiley, 1992), preferably in a suitable organic solvent or diluent, for example
DCM,
CHC13, THF, diethyl ether, n-heptane, n-hexane, n-pentane, cyclohexane,
diisopropyl ether, methyl tert-butyl ether, acetonitrile, DMF, DMSO, dioxane,
toluene, benzene, ethyl acetate or mixtures of these solvents, if appropriate
with
addition of a base such as, for example, butyllithium, lithium
diisopropylamide
(LDA), sodium hydride, sodium amide, potassium tent-butoxide, CaC03, Cs2C03,
triethylamine, diisopropylethylamine or complex bases (sodium amidelR~20Na,
where R~2 is (C2-Cs)-alkyl or CH3CH20CH2CH2), where for B = NRs an excess of
VII can also serve as a base.
a3) If appropriate, the protective groups in the compound of the formula VIII
on
R~ 3 andlor R~ ~ are removed by known methods (Greens, Wuts, Protective Groups
in Organic Synthesis,Wiley, 1991 ). If, for example, R~3 is a Boc-protected
amino
group, the Boc group can be removed, for example, by reaction with
trifluoroacetic
acid.
a4) If appropriate, R~3 in the compound of the formula VIII is then reacted
according to known processes to give the group D-E, for example by one of the
following processes.
a4.1 ) By reaction of compounds where R~3 = NHR6 with
1 H-pyrazole-1-carboxamidine or cyanamide a guanidine is obtained (see
Bernatowicz et al., J. Org. Chem. 57 (1992) 2497).
a4.2) By reaction of compounds where R~3 = NHRs with a monocyclic system or
polycyclic system of the type
CA 02225366 1997-12-19
N
N - _ L3
I6
R
5
in which L3 is a nucleophilically substitutable leaving group such as, for
example,
halogen or SH, SCH3, SOCH3, S02CH3 or HN-N02 , compounds with the end group
N
N "N
R6 I 6
are obtained (for the process see, for example, A.F. McKay et al., J. Med.
Chem. 6
(1963) 587; M.N. Buchman et al., J. Am. Chem. Soc. 71 (1949) 766; F. Jung et
al.,
J. Med. Chem. 34 (1991 ) 1110; or G. Sorba et al., Eur. J. Med. Chem. 21
(1986),
391 )
a4.3) By reaction of compounds where R~3 = NHR6 with compounds of the type
R6
~N
R6
\ N L3
6'
R
in which L3 is a nucleophilically substitutable leaving group such as, for
example,
halogen or SH, SCH3, SOCH3, S02CH3 or HN-N02, compounds with the end group
R6
\N
R\N~ N/
6'
R R6
CA 02225366 1997-12-19
41
are obtained (for the process see, for example, Miller, Synthesis 1986, 777;
or
Brimble, J. Chem. Soc., Perkin Trans. 1 (1990) 311 ).
a4.4) By reaction of compounds where R~3 = NHR6 with a monocyclic or
polycyclic
system of the type
N
N_ _L3
I6
R
in which L3 is a nucleophilically substitutable leaving group such as, for
example,
SCH3 , compounds with the end group
N
N "N
I6
R R6
are obtained (for the process see, for example, T. Hiroki et al., Synthesis
(1984)
703; or M. Purkayastha et al., Indian J. Chem. Sect. B 30 (1991 ) 646).
a 4.5) Compounds in which -D-E is the radical of an aminoguanidinylimine of
the
type
R6
~ N
6
R ~N ~N/N~ CR6
R6~ ~ 6
or of a cyclic aminoguanidinylimine of the type
CA 02225366 1997-12-19
42
N
~ N~CR6
N N
I6
R R6
can be prepared, for example, by condensation of compounds of the formula
R6
~N _N
g ~ or NH2
R ~N~ NH2 N
N~
~6'
6 6
with ketones or aldehydes of the type 0=C(Rs)- or corresponding acetals or
ketals
according to customary literature processes, for example analogously to N.
Desideri
et al., Arch. Pharm. 325 (1992) 773-777 or A. Alves et al., Eur. J. Med. Chem.
Chim.
Ther. 21 (1986) 297-304, where the above aminoguanidinylimines may be obtained
as EIZ isomer mixtures which can be separated by customary chromatographic
methods.
a4.6) Compounds in which -D-E is Rs-C(=NRs)-NR6-N=C(Rs)- or a radical
containing a monocyclic system or a polycyclic system, of the type
R6 R6
N~N
can be obtained analogously to a4.5).
a4.7) Compounds in which -D- is -S(0)2NR6- can be prepared, for example, by
oxidizing compounds with R~3 = SH to sulfonic acids (R~3 = S03H) by methods
known from the literature (cf. Houben-Weyl, Methoden der Organischen Chemie,
Bd. E1212, Georg Thieme Verlag, Stuttgart 1985, p. 1058ff), from which the
CA 02225366 1997-12-19
43
compounds with -D- _ -S(O)2NR6- are then prepared, for example, directly or
via
corresponding sulfonyl halides by linkage of an amide bond, where oxidation-
sensitive groups in the molecule, such as, for example, amino groups, amidino
groups or guanidino groups, are protected by suitable protective groups if
necessary
before carrying out the oxidation.
a4.8) Compounds in which -D- is -S(O)NR6- can be prepared, for example, by
converting compounds with R~3 = SH into the corresponding sulfide and then
oxidizing with meta-chloroperbenzoic acid to the sulfinic acids (R~3 = S02H)
(cf.
Houben-Weyl, Methoden der Organischen Chemie, Vol. E11I1, Georg Thieme
Verlag, Stuttgart 1985, p. 618f), from which the corresponding sulfinamides
can be
prepared according to methods known from the literature. Generally, other
methods
known from the literature can also be used for the preparation of compounds of
the
formulae I and la with -D- _ -S(O)~NR6- (u = 1, 2) (cf. Houben-Weyl, Methoden
der
Organischen Chemie, Vol. E11/1, Georg Thieme Verlag, Stuttgart 1985, p. 618ff
or
Vol. E1112, Stuttgart 1985, p. 1055ff).
a4.9) Compounds in which -D-E is RsRs~N-C(=NR6)-NRs-C(O)- or the radical of a
cyclic acylguanidine of the type
N
"N
R6 I 6
R
can be prepared, for example, by reacting a compound, in which R~ 3 is -C(O)-
L4
and L4 is an easily nucleophilically substitutable leaving group, with the
appropriate
guanidine (derivative) of the type
CA 02225366 1997-12-19
44
R6
\N
R6
\N NH
R6 ~ 6
R
or the cyclic guanidine (derivative) of the type
N
. N ~NH
16 16
R R
The activated acid derivatives with the group L4(O)C- indicated above, in
which L4
can be, for example, an alkoxy group, preferably a methoxy group, a phenoxy
group,
phenylthio group, methylthio group, 2-pyridylthio group or a nitrogen
heterocycle,
preferably 1-imidazolyl, are advantageously obtained in a manner known per se
from the carboxylic acid chlorides on which they are based (L4 = CI), which
for their
part can in turn be prepared in a manner known per se from the carboxylic
acids on
which they are based, for example using thionyl chloride. In addition to the
carboxylic acid chlorides (L4 = CI) further activated acid derivatives with
the group
of the type L4(O)C- can also be prepared in a known manner directly from the
carboxylic acids on which they are based (L4 = OH), such as, for example, the
methyl esters (L4 = OCH3) by treating with gaseous HCI in methanol, the
imidazolides (L4 = 1-imidazolyl) by treating with carbonyldiimidazole (cf.
Staab,
Angew. Chem. Int. Ed. Engl. 1, 351-367 (1962)) or the mixed anhydrides (L4 =
C2H50C(O)O or TosO) with CI-COOC2H5 or tosyl chloride in the presence of
triethylamine in an inert solvent. The activation of the carboxylic acids can
also be
carried out using carbodiimides like dicyclohexylcarbodiimide (DCCI) or using
O-
((cyano(ethoxycarbonyl)methylene)amino)-1,1,3,3-tetramethyluronium
tetrafluoroborate ("TOTU") (Konig et al., Proc. 21 st Europ. Peptide Symp.
1990,
(Eds. Giralt, Andreu), Escom, Leiden, 1991, p. 143) and other activating
reagents
CA 02225366 1997-12-19
customary in peptide chemistry (a number of suitable methods for the
preparation of
activated carboxylic acid derivatives are given with source literature in J.
March,
Advanced Organic Chemistry, Third Edition (John Wiley & Sons, 1985), p. 350).
The
reaction of an activated carboxylic acid derivative having the group of the
type
5 L4(O)C- with the respective guanidine (derivative) is preferably carried out
in a
manner known per se in a protic or aprotic polar, inert organic solvent, the
reaction
of the methyl esters (L4 = OMe) with the respective guanidines advantageously
being carried out in methanol, isopropanol or THF at temperatures from
20°C up to
the boiling temperature of these solvents. Most reactions of compounds having
the
10 group L4(O)C- with salt-free guanidines are advantageously carried out in
aprotic
inert solvents such as THF, dimethoxyethane or dioxane, it also being
possible,
however, to use water as a solvent when using a base such as, for example,
NaOH
in the reaction of compounds having the group L4(O)C- with guanidines. If L4 =
CI,
the reaction is advantageously carried out with the addition of an acid
scavenger, for
15 example in the form of excess guanidine (derivative), for the binding of
the
hydrohalic acid.
a4.10) Compounds in which -D-E is Rs-C(=NRs)-NRs-C(O)- or a radical comprising
a monocyclic system or polycyclic system, of the type
~N
16
R
can be obtained analogously to a4.9).
a4.11 ) Compounds in which -D-E is the radical of a sulfonylguanidine or
sulfoxylguanidine of the type RsRs~N-C(=NRs)-NR6-S(O)S- (u = 1, 2) or
CA 02225366 1997-12-19
46
N
~ / S(o)
N (u = 1, 2)
Rs Rs
can be prepared by methods known from the literature by reaction of compounds
of
the formulae RsRs~N-C(=NRs)-NHR6 and
N
"NH
R6 I 6
with compounds in which R~3 is S(0)~-L5 (u = 1, 2) and L5 is, for example, CI
or
NH2, for example analogously to S. Birtwell et al., J. Chem. Soc. (1946) 491
or
Houben Weyl, Methoden der Organischen Chemie, Vol. E4, Georg Thieme Verlag,
Stuttgart 1983; p. 620 ff.
a4.12) Compounds in which -D-E is R6-C(=NRs)NRs-S(O)u (u = 1, 2) or the
radical
comprising a monocyclic system or polycyclic system, of the type
/ S(0)~
Rs
(u= 1, 2), can be obtained analogously to a4.11 ).
a4.13) Compounds in which -D- is -NRs-C(O)- can be prepared, for example, by
first reacting a compound with R~3 = -NHRs with a suitable carbonic acid
derivative,
preferably phosgene, diphosgene (trichloromethyl chloroformate), triphosgene
(bis-
trichloromethyl carbonate), ethyl chloroformate, i-butyl chloroformate, bis(1-
hydroxy-
1 H-benzotriazolyl) carbonate or N,N'-carbonyldiimidazole, in a solvent which
is inert
to the reagents used, preferably DMF, THF or toluene, at temperatures between
CA 02225366 1997-12-19
47
20°C and the boiling point of the solvent, preferably between
0°C and 60°C, to give
a compound in which R~ 3 is
L6 NRs
where L6, depending on the carbonic acid derivative used, is, for example, a
hydroxyl group, halogen such as, for example, chlorine, ethoxy, isobutoxy,
benzotriazol-1-oxy or 1-imidazolyl. The subsequent reaction of these
derivatives
with R6Rs~N-C(=NRs)-NR6~H or Rs-C(=NRs)-NHRs or with the compound comprising
a monocyclic system or polycyclic system, of the type
N
~NH or HN
I I ~6
R6 R6
is then carried out as described above for the preparation of acylguanidines
(or their
derivatives) in a4.9).
a4.14) Compounds of the formula I in which -D-E is a bis-aminotriazole radical
or a
bis-aminooxadiazole radical can be prepared, for example, according to P.J.
Garrett
et al., Tetrahedron 49 (1993) 165 or R. Lee Webb et al., J. Heterocyclic Chem.
24
(1987) 275.
a4.15) Compounds of the formula I in which -D-E is a urea group or a thiourea
group can be synthesized according to known methods, such as are summarized,
for example, in C. Ferri, Reaktionen der organischen Synthese, Georg Thieme
Verlag, Stuttgart 1978, for example by reaction of the corresponding amines
with
isocyanates or isothiocyanates.
a5) If appropriate, after the reaction of R~3 in the compound of the formula
VIII to
CA 02225366 1997-12-19
48
give the group D-E, further protective groups still to be removed are removed
by
known methods (see Greene, Wuts, see above).
a6) If appropriate, the compounds of the formula I obtained are converted into
their salts, in particular into pharmaceutically utilizable or nontoxic,
physiologically
tolerable salts, and/or are converted into prodrugs.
The present invention furthermore also relates to processes for the synthesis
of the
compounds of the formula la, which comprise carrying out one or more of the
following steps for the synthesis of the compounds of the formula la.
b1 ) A compound of the formula IV is reacted with a compound of the formula
IX,
L2-(CRS R2)~ A'-(CRS R2)s (CRS R3)k-(CRS R2)t-R13 (IX);
in which R~, R2, R3, A', r, s, k, t, R~3 and L2 are as defined above,
to give a compound of the formula X
1
N
~~-Y (X)
N
X N
R~s
in which R~s is -(CRS R2)~ A'-(CRS R2)S (CRS R3)k-(CRS R2)t-R~3, L1, X and Y
are as
defined above and the meanings indicated above otherwise apply. The reaction
is
carried out by methods known to those skilled in the art (see source
literature in J.
March, Advanced Organic Chemistry, Fourth Edition, Wiley, 1992), preferably in
a
suitable organic solvent or diluent, for example DCM, CHC13, THF, diethyl
ether,
n-heptane, n-hexane, n-pentane, cyclohexane, diisopropyl ether, methyl tert-
butyl
ether, acetonitrile, DMF, DMSO, dioxane, toluene, benzene, ethyl acetate or
mixtures of these solvents, if appropriate with addition of a base such as,
for
CA 02225366 1997-12-19
49
example, butyllithium, lithium diisopropylamide (LDA), sodium hydride, sodium
amide, potassium tart-butoxide, CaC03, Cs2C03, triethylamine,
diisopropylethylamine or complex bases (sodium amidelR~20Na, where R~2 is (C2-
C6)-alkyl or CH3CH20CH2CH2). For L2 = OH, the reaction can be carried out, for
example, by the conditions described for the Mitsunobu reaction (Hughes,
Organic
Reactions 42 (1992) 335-656), for example by reaction with triphenylphosphine
and
DEAD in THF.
b2) The compound of the formula X is reacted with a compound of the formula
XI
H-B-(CR~R2)~-A-(CR~R2)m-(CR~R3)~ (CR~R2)q-R1o (XI);
in which R~, R2, R3, R~~, A, B, n, m, i and q are as defined above,
to give a compound of the formula XII
R"
N ~ N~Y (X11)
N
X N
Rig
in which R~6, X and Y are as defined above, R~~ is -B-(CR~R2)~-A-(CR~R2)m-
(CRS R3)~ (CRS R2)q-R~ ~ and the meanings indicated above otherwise apply.
b3) For the further synthesis of compounds of the formula la, the procedure is
then analogous to the steps a3) to a6) in the synthesis of the compounds of
the
formula I.
In the process for the synthesis of the compounds of the formula I, step a2)
can also
be carried out before a1 ). In the process for the synthesis of the compounds
of the
formula la, step b2) can also be carried out before b1 ).
CA 02225366 1997-12-19
The introduction of carbon substituents in the 6-position of the purine
structure can
be carried out, for example, by Stille coupling, such as described, for
example, in
Langli et al., Tetrahedron 52 (1996) 5625; Gundersen, Tetrahedron Lett. 35
(1994)
3153, or by Heck coupling, such as described, for example, in Koyama et al.,
5 Nucleic Acids Res., Symp. Ser. 11 (1982) 41.
A substituent X in position 2 of the purine structure can also be introduced
by known
methods at the end of the synthesis of the compounds of the formulae I and la,
such
as described, for example, in D. A. Nugiel, J. Org. Chem. 62 (1997) 201-203;
N. S.
10 Gray, Tetrahedron Lett. 38 (1997) 1161 and the references cited there.
A substituent representing Y in the 8-position can be introduced by known
methods,
such as described, for example, in E. J. Reist et al., J. Org. Chem. 33 (1968)
1600;
J. L. Kelley et al., J. Med. Chem. 33 (1990) 196; or E. Vanotti et al., Eur.
J. Chem.
15 29 (1994) 287.
The compounds of the formulae I and la according to the invention and their
physiologically tolerable salts can be administered to animals, preferably to
mammals, and in particular to humans as pharmaceuticals per se, in mixtures
with
20 one another or in the form of pharmaceutical preparations which permit
enteral or
parenteral use and which, as active constituent, contain an efficacious dose
of at
least one compound of the formula I or of the formula la or of a salt thereof
or of a
prodrug thereof in addition to customary pharmaceutically innocuous vehicles
andlor additives. The pharmaceutical preparations normally contain
approximately
25 0.5 to 90°~ by weight of the therapeutically active compounds.
The pharmaceuticals can be administered orally, for example in the form of
pills,
tablets, lacquered tablets, coated tablets, granules, hard and soft gelatin
capsules,
solutions, syrups, emulsions, suspensions or aerosol mixtures. Administration,
30 however, can also be carried out rectally, for example in the form of
suppositories,
or parenterally, for example in the form of injection solutions or infusion
solutions,
microcapsules or rods, percutaneously, for example in the form of ointments or
tinctures, or nasally, for example in the form of nasal sprays.
CA 02225366 1997-12-19
51
The pharmaceutical preparations are prepared in a manner known per se,
pharmaceutically inert inorganic or organic vehicles being used. For the
production
of pills, tablets, coated tablets and hard gelatin capsules, it is possible to
use, for
example, lactose, corn starch or derivatives thereof, talc, stearic acid or
its salts etc.
Vehicles for soft gelatin capsules and suppositories are, for example, fats,
waxes,
semisolid and liquid polyols, natural or hardened oils, etc. Suitable vehicles
for the
production of solutions and syrups are, for example, water, sucrose, invert
sugar,
glucose, polyols, etc. Suitable vehicles for the production of injection
solutions are
water, alcohols, glycerol, polyols, vegetable oils, etc. Suitable vehicles for
microcapsules, implants or rods are copolymers of glycolic acid and lactic
acid.
In addition to the active compounds and vehicles, the pharmaceutical
preparations
can additionally contain additives, such as, for example, fillers, extenders,
disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers,
preservatives, sweeteners, colorants, flavorings or aromatizers, thickeners,
diluents,
buffer substances, and also solvents or solubilizers or agents for achieving a
depot
effect, and also salts for altering the osmotic pressure, coating agents or
antioxidants. They can also contain two or more compounds of the formula I or
la
andlor their physiologically tolerable salts, and also, in addition to at
least one
compound of the formula I or la or of a salt thereof, additionally one or more
other
therapeutically active substances.
The dose can vary within wide limits and is to be suited to the individual
conditions
in each individual case. In the case of oral administration, the daily dose is
in
general approximately 0.01 to 100 mglkg, preferably 0.1 to 5 mg/kg, in
particular 0.3
to 0.5 mglkg, of body weight to achieve effective results. Also in the case of
intravenous administration the daily dose is in general approximately 0.01 to
100
mglkg, preferably 0.05 to 10 mglkg, of body weight. The daily dose can be
divided,
in particular in the case of the administration of relatively large amounts,
into
several, for example 2, 3 or 4, part administrations. If appropriate,
depending on the
individual behavior, it may be necessary to deviate upwards or downwards from
the
daily dose indicated.
CA 02225366 1997-12-19
52
Apart from use as pharmaceutical active compounds, the compounds of the
formulae I and la can also be employed for diagnostic purposes, for example in
in
vitro diagnoses, and as tools in biochemical investigations in which
inhibition of the
vitronectin receptor or influencing of cell-cell or cell-matrix interactions
is intended.
Furthermore, they can be used as intermediates for the preparation of other
compounds, in particular of other pharmaceutical active compounds, which are
obtainable from the compounds of the formulae I and la, for example, by
modification or introduction of residues or groups.
Abbreviations
used:
AcOH acetic acid
Boc tert-butoxycarbonyl
DCCI dicyclohexylcarbodiimide
DCM dichloromethane
DEAD diethyl azodicarboxylate
DIPEA diisopropylethylamine
DMF dimethylformamide
DMSO dimethyl sulfoxide
EA ethyl acetate
HOOBt 3-hydroxy-4-oxo-3,4-dihydro-1,2,3-benzotriazine
MeOH methanol
Mes methylsulfonyl
RT room temperature
Tf trifluoromethylsulfonyl
THF tetrahydrofuran
Tos p-toluenesulfonyl
Z benzyloxycarbonyl
CA 02225366 1997-12-19
53
Examples
Compounds of the formulae I and la which in the 6-position of the purine
structure
contain an amino group which is not a constituent of a ring can also be
regarded as
derivatives of adenine (= 6-aminopurine) and can be designated as such in the
nomenclature of the compounds. Substituents which are bonded to the nitrogen
atom of the amino group in the 6-position of adenine are provided in this
notation
with the addendum Ns. Substituents which are bonded to the ring nitrogen atom
in
the 9-position are provided with the addendum N9. In the name of the
substituent it
is indicated at the beginning via which position in the substituent the
substituent is
bonded to the nitrogen atom N6 or N9 in the chosen notation. The same applies
to
compounds which are designated as N9-substituted derivatives of purine.
NHZ
7
1 N 6 ~ 5 N 7 ~ N \ N> 8 Purine
~> 8 Adenine 2 ~ N~4 g
2 N 4 N9 3
H3
Example 1
Ns-(1-(5-Guanidinopentyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic acid))-
adenine
1 a) N9-(3-(tert-Butyl 2S-(benzyloxycarbonylamino)propionate))-6-chloropurine
2.63 g (17 mmol) of 6-chloropurine and 4.46 g (16.5 mmol) of
triphenylphosphine
were suspended in 50 ml of absol. THF under argon. 2.56 ml (16.3 mmol) of DEAD
were added at RT and the mixture was stirred at RT for 15 minutes, a clear
solution
being formed. 3.78 g (12.8 mmol) of N-benzyloxycarbonyl-L-serine tert-butyl
ester
(prepared according to M. Schultz, H. Kunz, Tetrahedron: Asymmetry 4 (1993)
1205-1220), dissolved in 50 ml of absol. THF, were added to this solution
during the
CA 02225366 1997-12-19
54
course of 1.5 h. The mixture was then stirred at RT for a further 2 h. The
solvent
was evaporated, and the residue was triturated with ether and chromatographed
through silica gel (toluene:EA 98:2 to 7:3), 2.85 g (51°~) of pure
product being
obtained.
~H-NMR (200 MHz, DMSO): 8 = 1.30 (s, 9H, C(CH3)3); 4.48-4.73 (m, 3H, N9-CH2-
CH(NHZ)); 4.98 (s, 2H, CH2-aryl); 7.19-7.40 (m, 5H, aryl-H); 7.87 (d, 1 H,
NH); 8.61
+ 8.77 (2 s, 2H, Cs-H + C8-H).
MS (FAB): m/e = 432.1 (100%; (M+H)+); 376.0 (60).
1b) N6-(1-(5-(tert-Butyloxycarbonylamino)pentyl))-N9-(3-(tart-butyl
2S-(benzyloxycarbonylamino)propionate))-adenine
0.170 ml (1 mmol) of DIPEA and 5 mg of potassium iodide were added to a
solution
of 431 mg (1 mmol) of N9-(3-(tart-butyl 2S-
(benzyloxycarbonylamino)propionate))-6-
chloropurine (Example 1 a) and 404 mg (2 mmol) of 5-(tert-
butyloxycarbonylamino)-
1-pentylamine in 5 ml of absol. DMF and the mixture was stirred at 40°C
for 72 h.
The solvent was evaporated and the residue was chromatographed through silica
gel (toluene:EA 7:3 to 1:2), 190 mg (32°~) of pure product being
obtained.
MS (FAB): mle = 598.3 (100°~; (M+H)+).
1c) N6-(1-(5-Aminopentyl))-N9-(3-(2-(benzyloxycarbonylamino)propionic acid))-
adenine
190 mg (0.32 mmol) of Ns-(1-(5-(tert-butyloxycarbonylamino)pentyl))-N9-(3-
(tert-
butyl 2S-(benzyloxycarbonylamino)propionate))-adenine (Example 1 b) were
dissolved in 2 ml of 90°~ strength trifluoroacetic acid and the
solution was stirred at
RT for 2 h. It was evaporated to dryness and the residue was coevaporated
twice
with acetic acid. The residue was then dissolved in water and freeze-dried.
Yield:
134 mg (95%).
MS (ES+): mle = 442.3 (20%; (M+H)+), 308.2 (35).
1d) Ns-(1-(5-Guanidinopentyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic
acid))-adenine
CA 02225366 1997-12-19
34 mg (0.077 mmol) of N6-(1-(5-aminopentyl))-N9-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-adenine (Example 1 c) were dissolved
in
1.5 ml of water and 0.5 ml of DMF and the solution was treated with 0.033 ml
(0.193
mmol) of DIPEA and 13.5 mg (0.092 mmol) of 1H-pyrazole-1-carboxamidine
5 hydrochloride and stirred at RT for 40 h. The solvent was then evaporated,
the
residue was taken up in water and the solution was freeze-dried. For further
purification, it was chromatographed through silica gel (DCM:methanol:acetic
acid:water 15:5:1:1 ). Yield: 70°~.
MS (FAB): m/e = 484.2 (100%; (M+H)+)
Example 2
Ns-(1-(4-Guanidinobutyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic acid))-
adenine
2a) N6-(1-(4-(tert-Butyloxycarbonylamino)butyl))-N9-(3-(tert-butyl 2S-
(benzyloxycarbonylamino)propionate))-adenine
Synthesis analogously to 1 b from 431 mg (1 mmol) of N9-(3-(tert-butyl 2S-
(benzyloxycarbonylamino)propionate))-6-chloropurine (Example 1 a) and 376 mg
(2
mmol) of 4-(tart-butyloxycarbonylamino)-1-butylamine. Yield: 214 mg
(37°~).
~H-NMR (200 MHz, DMSO): 8 = 1.30 (s, 9H, C(CH3)3); 1.38 (s, 9H, C(CH3)3); 1.41
(m, 2H, CH2); 1.57(m, 2H, CH2); 3.46 (m, 2H, CH2-NH-Boc); 2.92 (t, 2H, C2-NH-
CH2); 4.31-4.58 (m, 3H, N~-CH2-CH(NHZ)); 5.01 (s, 2H, CH2-aryl); 6.99 (t, 1H,
C2-
NH); 7.10-7.38 (m, 5H, aryl-H); 7.75 (m, 1 H, NH-Boc); 7.91 (d, 1 H, NH-Z);
8.02 + 8.20 (2 s, 2H, C6-H + C8-H).
MS (ES+): mle = 584.3 (100%; (M+H)+)
2b) Ns-(1-(4-Aminobutyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic acid))-
adenine
Synthesis analogously to Example 1 c from Ns-(1-(4-tert-
butyloxycarbonylamino)butyl)-N9-(3-(tert-butyl 2S-
CA 02225366 1997-12-19
56
(benzyloxycarbonylamino)propionate)-adenine (Example 2a). Yield: 96%.
MS (ES+): m/e = 428.2 (100°~; (M+H)+), 294.1 (70).
2c) N6-(1-(4-Guanidinobutyl))-N9-(3-(2S-(benzyloxycarbonylamino)-propionic
acid))-
adenine
Synthesis analogously to Example 1d from N6-(1-(4-aminobutyl))-N9-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-adenine (Example 2b). Yield: 76%.
MS (ES+): mle = 470.1 (20°~; (M+H)+).
Example 3
N6-(1-(3-Guanidinopropyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic acid))-
adenine
3a) N6-(1-(3-(tart-Butyloxycarbonylamino)propyl))-N9-(3-(tert-butyl 2S-
(benzyloxycarbonylamino)propionate))-adenine
Synthesis analogously to 1 b from 60 mg (0.14 mmol) of N9-(3-(tart-butyl 2S-
(benzyloxycarbonylamino)propionate))-6-chloropurine (Example 1 a) and 30 mg
(0.17 mmol) of 3-(tert-butyloxycarbonylamino)-1-propylamine. Yield: 30 mg
(38°~).
~H-NMR (200 MHz, DMSO): 8 = 1.28 (s, 9H, C(CH3)3); 1.36 (s, 9H, C(CH3)3); 1.68
(m, 2H, CH2-CH2-CH2); 1.41 (m, 2H, CH2); 2.98 (t, 2H, C2-NH-CH2); 3.46 (t, 2H,
CH2-NH-Boc); 4.29-4.59 (m, 3H, N~-CH2-CH(NHZ)); 5.00 (s, 2H, CH2-aryl); 6.82
(t,
1 H, C2-NH); 7.21-7.40 (m, 5H, aryl-H); 7.72 (m, 1 H, NH-Boc); 7.91 (d, 1 H,
NH-Z);
8.03 + 8.20 (2 s, 2H, C6-H + C8-H).
MS (ES+): mle = 570.3 (100%; (M+H)+)
3b) Ns-(1-(3-Aminopropyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic acid))-
adenine
Synthesis analogously to Example 1 c from N6-(1-(3-(tert-
butyloxycarbonylamino)propyl))-N9-(3-(tert-butyl 2S-
CA 02225366 1997-12-19
57
(benzyloxycarbonylamino)propionate))-adenine (Example 3a). Yield: 100%.
MS (ES+): mle = 414.2 (100°~; (M+H)+), 280.1 (30).
3c) N6-(1-(3-Guanidinopropyl))-N9-(3-(2S-(benzyloxycarbonylamino)propionic
acid))-adenine
Synthesis analogously to Example 1d from N6-(1-(3-aminopropyl))-N9-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-adenine (Example 3b). Yield: 66%.
MS (ES+): mle = 456.3 (20°~; (M+H)+), 130.1 (100).
Example 4
N6-(1-(4-(4,5-Dihydro-1 H-imidazol-2-ylamino)butyl))-N9-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-adenine
153 mg (0.36 mmol) of N6-(1-(4-aminobutyl))-N9-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-adenine (Example 2b) and 88 mg (0.36
mmol) of 2-(methylmercapto)-2-imidazoline hydroiodide were dissolved in 2 ml
of
water and the solution was adjusted to pH 9.0 using 1 N NaOH. It was stirred
at
50°C for 100 h. The solution was then brought to pH 1.5 using 1 N HCI,
the solvent
was evaporated and the residue was chromatographed several times through
silica
gel (DCM: MeOH 9:1 to 1:2, in each case using 0.1 °~ AcOH, 0.1
°~ H20), then
DCM:MeOH:H20:AcOH 8:2:0.4:0.4. Yield: 7 mg (4%).
MS (FAB): mle = 496.2 (M+H+, 100%); 518.2 (M+Na+, 50).
Example 5
Ns-(1-(3-Guanidinopropyl))-N9-(4-(2S-(benzyloxycarbonylamino)butyric acid))-
adenine
5a) N9-(4-(tert-Butyl 2S-(benzyloxycarbonylamino)butyrate))-6-chloropurine
Synthesis analogously to Example 1 a from 6-chloropurine and N-benzyl-
CA 02225366 1997-12-19
58
oxycarbonyl-L-homoserine tert-butyl ester. Yield: 24%.
~ H-NMR (200 MHz, DMSO): 8 = 1.34 (s, 9H, C(CH3)3); 2.08-2.43 (m, 2H, N-CH2-
CH2-CH); 3.81-3.93(m, 1 H, CH-NHZ); 4.39 (t, 2H, N9-CH2); 5.02 (s, 2H, CHZ-
aryl);
7.26-7.42 (m, 5H, aryl-H); 7.87 (d, 1 H, NH); 8.63 + 8.75 (2 s, 2H, C6-H + C8-
H).
MS (FAB): mle = 446.1 (100%; (M+H)+); 390.1 (65).
5b) Ns-(1-(3-(tart-Butyloxycarbonylamino)propyl))-N9-(4-(tart-butyl 2S-
(benzyloxycarbonylamino)butyrate))-adenine
Synthesis analogously to 1 b from 50 mg (0.11 mmol) of N9-(4-(tert-butyl 2S-
(benzyloxycarbonylamino)butyrate))-6-chloropurine (Example 5a) and 38 mg (0.22
mmol) of 3-(tert-butyloxycarbonylamino)-1-propylamine. Yield: 26 mg (41
°~).
MS (ES+): m/e = 584.3 (100°~; (M+H)+).
5c) Ns-(1-(3-Aminopropyl))-N9-(4-(2-(benzyloxycarbonylamino)butyric acid))-
adenine
Synthesis analogously to Example 1 c from Ns-(1-(3-(tert-
butyloxycarbonylamino)propyl))-N9-(4-(tart-butyl 2S-(benzyloxycarbonylamino)-
butyrate))-adenine (Example 5b). Yield: 94°~.
MS (FAB): mle = 428.3 (100%; (M+H)+).
5d) N6-(1-(3-Guanidinopropyl))-N9-(4-(2S-(benzyloxycarbonylamino)butyric
acid))-
adenine
Synthesis analogously to Example 1d from N6-(1-(3-aminopropyl))-N9-(3-(2S-
(benzyloxycarbonylamino)butyric acid))-adenine (Example 5c). Yield: 71 %.
MS (FAB): mle = 470.3 (70%; (M+H)+).
5e) N-Benzyloxycarbonyl-L-homoserine
6 g (50.4 mmol) of L-homoserine were largely dissolved in 50 ml of DMF and
treated
at 0°C in portions with 12.56 g (50.4 mmol) of N-
(benzyloxycarbonyloxy)succinimide.
The mixture was stirred at 0°C for 1 h, then at RT for 48 h. The
solvent was distilled
CA 02225366 1997-12-19
59
off and the residue was partitioned between EA and a saturated NaCI solution.
The
organic phase was washed with saturated NaCI solution, with 5°~
strength citric acid
and again with saturated NaCI solution, dried, filtered and concentrated. The
crystalline residue was stirred in ether, filtered off with suction, and
washed with
ether and pentane. Yield: 9.55 g (75°~).
~H-NMR (200 MHz, DMSO): 8 = 1.61-1.95 (m, 2H, CH2-CH2-OH); 3.42 (m, 2H, CH2-
OH); 4.08 (m, 1 H, CH-NH-Z); 4.57 (s, broad, 1 H, OH); 5.02 (s, 2H, CH2-Ph);
7.32
(m, 5H, aryl-H), 7.49 (d, 1 H, NH-Z).
MS (CI+): mle = 236.1 (M+H+-H20, 20°~); 192.1 (50); 91.0 (100).
5f) N-Benzyloxycarbonyl-L-homoserine tart-butyl ester
3.8 g (15 mmol) of Z-L-homoserine and 3.42 g (15 mmol) of benzyl-
triethylammonium chloride were dissolved under argon in 110 ml of N-methyl-2-
pyrrolidone and treated successively with 53.9 g (390 mmol) of K2C03 and 98.7
g
(720 mmol) of tert-butyl bromide. The solution was stirred at 55°C for
22 h. The
reaction mixture was poured into 1.5 I of ice water, extracted twice with
toluene, and
the organic phase was washed twice with saturated NaCI solution, dried,
filtered and
concentrated. The product was chromatographed through silica gel for further
purification (n-heptane : EA 7:3 to 1:1 ). Yield: 2.0 g (43.1 %).
~H-NMR (200 MHz, CDC13): 8 = 1.45 (s, 9H, tBu); 1.51-1.74 + 2.03-2.26 (m, 2H,
CH2-CH2-OH); 3.01 (s, broad, 1 H, OH); 3.70 (m, 2H, CH2-OH); 4.41 (m, 1 H, CH-
NH-
Z); 5.12 (s, 2H, CH2-Ph); 5.60 (d, 1 H, NH-Z); 7.36 (m, 5H, aryl-H),.
MS (CI+): m/e = 310.3 (M+H+, 50°~); 254.2 (100).
Example 6
Ns-(1-(4-Guanidinobutyl))-N9-(4-(2S-(benzyloxycarbonylamino)butyric acid))-
adenine
6a) N6-(1-(4-(tert-Butyloxycarbonylamino)butyl)-N9-(4-(tent-butyl 2S-
(benzyloxycarbonylamino)butyrate))-adenine
CA 02225366 1997-12-19
Synthesis analogously to 1 b from 50 mg (0.11 mmol) of N9-(4-(tert-Butyl 2S-
(benzyloxycarbonylamino)butyrate))-6-chloropurine (Example 5a) and 41 mg (0.22
mmol) of 4-(tert-butyloxycarbonylamino)-1-butylamine. Yield: 38 mg
(58°~).
MS (ES+): mle = 598.3 (100°~; (M+H)+).
5
6b) Ns-(1-(4-Aminobutyl))-N9-(4-(2-(benzyloxycarbonylamino)butyric acid))-
adenine
Synthesis analogously to Example 1 c from N6-(1-(4-(tert-
butyloxycarbonylamino)butyl))-N9-(4-(tert-butyl 2S-(benzyloxycarbonylamino)-
10 butyrate))-adenine (Example 6a). Yield: 100°~.
MS (FAB): mJe = 442.3 (100%; (M+H)+).
6c) N6-(1-(4-Guanidinobutyl))-N9-(4-(2S-(benzyloxycarbonylamino)butyric acid))-
adenine
Synthesis analogously to Example 1d from Ns-(1-(4-aminobutyl))-N9-(3-(2S-
(benzyloxycarbonylamino)butyric acid))-adenine (Example 6b). Yield:
65°~.
MS (ES+): mle = 484.3 (5%; (M+H)+), 350.2 (10), 333.2 (5), 130.0 (100).
Example 7
Ns-(1-(3-Guanidinopropyl))-N9-(3-propionic acid)-adenine
7a) N9-(tent-butyl 3-propionate)-6-chloropurine
15.45 g (0.1 mol) of 6-chloropurine, 43.5 ml (0.3 mol) of tert-butyl acrylate
and 1.34
ml (7 mmol) of 5.22 N sodium methoxide (in MeOH) were dissolved in 400 ml of
absol. MeOH and boiled under reflux for 4.5 h with repeated addition of 2.6 ml
(14
mmol) of 5.22 N sodium methoxide (in MeOH). For working-up, the solid was
filtered
off with suction, the solvent was evaporated and the residue was
chromatographed
(toluene:EA 3:1 ) through silica gel (+10% H20). Yield: 1.35 g (5%).
~ H-NMR (200 MHz, DMSO): 8 = 1.29 (s, 9H, C(CH3)3); 2.95 (t, 2H, CH2C(O));
4.50
(t, 2H, N-CH2); 8.70 + 8.79 (2 s, 2H, C6-H + C8-H).
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61
MS (ES+): mle = 283.1 (70%; (M+H)+); 227.0 (100).
7b) Ns-(1-(3-(tart-Butyloxycarbonylamino)propyl))-N9-(tart-butyl 3-propionate)-
adenine
Synthesis analogously to 1 b from 282 mg (1.0 mmol) of N9-(tart-butyl 3-
propionate)-
6-chloropurine (Example 7a) and 209 mg (1.2 mmol) of 3-(tert-
butyloxycarbonylamino)-1-propylamine. Yield: 160 mg (38°r6).
MS (ES+): m/e = 421.2 (100%; (M+H)+), 365.2 (60), 321.2 (50), 265.1 (30).
7c) Ns-(1-(3-Aminopropyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example 1cfrom N6-(1-(3-(tert-
butyloxycarbonylamino)propyl))-N9-(tart-butyl 3-propionate)-adenine (Example
7b).
YieId:100°~.
~ H-NMR (200 MHz, DMSO): 8 = 1.88 (t, 2H, CH2-CH2-CH2-); 2.80-2.93 (m, 4H, NH-
CH2 + CH2-C(O)); 3.56 (m, 2H, CH2-NH2); 4.38 (t, 2H, N9-CH2); 7.72 (s, broad,
2H,
NH2); 7.95 (t, 1 H, NH); 8.15 + 8.23 (2 s, 2H, Cs-H + C8-H).
MS (ES+): mle = 265.1 (100°~; (M+H)+); 248.1 (40), 176.0 (30).
7d) Ns-(1-(3-Guanidinopropyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example 1 d from Ns-(1-(3-aminopropyl))-N9-(3-
propionic
acid)-adenine (Example 7c). Yield: 41 °~.
~ H-NMR (200 MHz, D20): 8 = 1.95 (t, 2H, CH2-CH2-CH2-); 2.71 (t, 2H, CH2-
C(O));
3.24 (t, 2H, Gua-CH2); 3.65 (m, 2H, CH2-NH2); 4.40 (t, 2H, N9-CH2); 8.00 +
8.15 (2
s, 2H, C6-H + C8-H).
MS (ES+): mle = 307.1 (100°~; (M+H)+), 290.1 (30).
Example 8
N6-(1-(4-Guanidinobutyl))-N9-(3-propionic acid)-adenine
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62
8a) Ns-(1-(4-(tart-Butyloxycarbonylamino)butyl))-N9-(tert-butyl 3-propionate)-
adenine
Synthesis analogously to 1 b from 141 mg (0.5 mmol) of N9-(tert-butyl 3-
propionate)-
6-chloropurine (Example 7a) and 104 mg (0.55 mmol) of 4-(tert-
butyloxycarbonylamino)-1-butylamine. Yield: 130 mg (60°~).
~ H-NMR (200 MHz, DMSO): 8 = 1.32 (s, 9H, C(CH3)3); 1.35 (s, 9H, C(CH3)3);
1.40
(t, 2H, CH2); 1.57 (t, 2H, CH2); 2.84 (t, 2H, -CH2-C(O)); 2.95 (t, 2H, C2-NH-
CH2);
3.45 (m, 2H, CH2-NH-Boc); 4.34 (t, 2H, N9-CH2); 6.78 (t, 1 H, C2-NH); 7.70 (m,
1 H,
NH-Boc); 8.08 + 8.19 (2 s, 2H, Cs-H + C8-H).
MS (ES+): m/e = 435.2 (100°~; (M+H)+), 379.2 (20), 335.2 (55),
279.1 (50).
8b) N6-(1-(4-Aminobutyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example lcfrom Ns-(1-(4-(tert-
butyloxycarbonylamino)butyl)-N9-(tert-butyl 3-propionate)-adenine (Example
8a).
Yield: 100°~.
~ H-NMR (200 MHz, DMSO): 8 = 1.50-1.70 (m, 4H,-CH2-CH2-); 2.74-2.91 (m, 4H,
NH-CH2 + CH2-C(O)); 3.50 (m, 2H, CH2-NH2); 4.36 (t, 2H, N9-CH2); 7.64 (s,
broad,
2H, NH2); 7.90 (t, 1 H, NH); 8.11 + 8.21 (2 s, 2H, C6-H + C8-H).
MS (FAB): mle = 279.2 (100%; (M+H)+)
8c) N6-(1-(4-Guanidinobutyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example 1d from Ns-(1-(4-aminobutyl))-N9-(3-propionic
acid)-adenine (Example 8b). Yield: 65%.
MS (ES+): mle = 321.1 (100%; (M+H)+).
Example 9
Ns-(1-(5-Guanidinopentyl))-N9-(3-propionic acid)-adenine
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63
9a) N6-(1-(5-(tert-Butyloxycarbonylamino)butyl))-N9-(tert-butyl 3-propionate)-
adenine
Synthesis analogously to 1 b from 282 mg (1.0 mmol) of N9-(tart-butyl 3-
propionate)-
6-chloropurine (Example 7a) and 243 mg (1.2 mmol) of 5-(tert-
butyloxycarbonylamino)-1-pentylamine. Yield: 219 mg (41 °~).
MS (ES+): mle = 449.3 (100%; (M+H)+)
9b) Ns-(1-(5-Aminopentyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example 1c from N6-(1-(5-(tert-
butyloxycarbonylamino)pentyl))-N9-(tert-butyl 3-propionate)-adenine (Example
9a).
Yield: 100%.
~H-NMR (200 MHz, DMSO): b = 1.39 (m, 2H, CH2) 1.50-1.67 (m, 4H, 2 x CH2); 2.79
(dt, 2H, NH-CH2); 2.89 (m, 2H, CH2-C(O)); 3.48 (m, 2H, CH2-NH2); 4.37 (t, 2H,
N9-
CH2); 7.67 (s, broad, 2H, NH2); 8.04 (t, 1 H, NH); 8.13 + 8.25 (2 s, 2H, Cs-H
+ C8-H).
MS (ES+): m/e = 293.1 (100°~; (M+H)+).
9c) Ns-(1-(5-Guanidinopentyl))-N9-(3-propionic acid)-adenine
Synthesis analogously to Example 1d from Ns-(1-(5-aminopentyl))-N9-(3-
propionic
acid)-adenine (Example 9b). Yield: 37%.
~ H-NMR (200 MHz, DMSO): b = 1.38-1.79 (m, 6H, 3 x CH2); 2.80 (t, 2H, NH-CH2);
3.12 (m, 2H, CH2-C(O)); 3.58 (m, 2H, CH2-Gua); 4.43 (t, 2H, N9-CHZ); 8.07 +
8.21
(2 s, 2H, C6-H + C8-H).
MS (FAB): mle = 335.2 (100%; (M+H)+)
Example 10
Ns-(2-Acetic acid)-N9-(1-(5-aminopentyl))-adenine
10a) N6-(tert-Butyl 2-acetate)-adenine
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64
155 mg (1 mmol) of 6-chloropurine and 420 mg (2 mmol) of glycine tert-butyl
ester
hydrochloride (80°~ strength) were dissolved in 5 ml of absol. DMF and
treated with
0.17 ml of DIPEA and a spatula tipful of potassium iodide and the mixture was
stirred at 50°C for 6 h. The solvent was evaporated and the residue was
chromatographed through silica gel (toluene:EA 1:1 to 1:2). Yield: 76 mg
(31°~)
MS (ES+): 250.0 (M+H, 10%); 193.9 (95), 163.9 (100).
10b) N6-(2-Acetic acid)-N9-(1-(5-(tert-butyloxycarbonylamino)pentyl))-adenine
75 mg (0.3 mmol) of Ns-(tert-butyl 2-acetate)-adenine (Example 10a), 214 mg
(0.6
mmol) of 5-(tert-butyloxycarbonylamino)pentyl 4-toluene-sulfonate) and 42 mg
(0.3
mmol) of K2C03 were dissolved in 6 ml of absol. DMF and the solution was
stirred at
RT for 5 days. The solvent was evaporated and the residue was chromatographed
through silica gel (toluene:EA 7:3 to 1:2). Yield: 92 mg (71%).
MS (ES+): 435.3 (M+H, 25%); 349.3 (100).
10c) Ns-(2-Acetic acid)-N9-(1-(5-aminopentyl))-adenine
Synthesis analogously to Example 1 c from Ns-(2-acetic acid)-N9-(1-(5-(tert-
butyloxycarbonylamino)pentyl))-adenine (Example 10b). Yield: 93°~.
MS (ES+): mle = 279.2 (15°~; (M+H)+, 249.1 (100).
Example 11
Ns-(2-(N-(2-Aminoethyl)acetamide))-N9-(2-acetic acid)-adenine
11 a) N9-(tert-butyl 2-acetate)-adenine
6.76 g (0.05 mol) of -adenine were suspended in 300 ml of absol. DMF under N2,
then 2.4 g (0.06 mol) of NaH dispersion were added and the mixture was stirred
at
RT for 2 h. 14.7 ml (0.1 mol) of tert-butyl bromoacetate were added dropwise
in the
course of 30 min, a clear solution being formed. It was stirred at RT for a
further 5 h.
The solvent was evaporated, the residue was stirred with 500 ml of water, and
the
CA 02225366 1997-12-19
solid was filtered off with suction and crystallized from ethanol. Yield: 5.1
g (41 %).
~ H-NMR (200 MHz, DMSO): b = 1.42 (s, 9H, tBu); 4.95 (s, 2H, N9-CH2); 7.22 (s,
broad, 2H, NsH2); 8.10 + 8.15 (2 s, 2H, Cs-H + C8-H).
MS (ES+): m/e = 250.1 (M+H+, 65 %), 194.0 (100).
5
11 b) N6-(ethyl 2-acetate)-N9-(tert-butyl 2-acetate)-adenine
978 mg (3 mmol) of NaH and 250 mg (1 mmol) of N9-(tert-butyl 2-acetate)-
adenine
(Example 11 a) were suspended in 10 ml of absol. DMF and 0.12 ml of ethyl
10 chloroacetate was added dropwise during the course of 10 min. The mixture
was
then stirred at 50°C for 6 h, then the same amount of CsC03 was added
again and
the mixture was stirred at 50°C for 6 h. The solvent was evaporated and
the residue
was partitioned between water and EA. The organic phase was dried and
concentrated. Yield: 16°~.
15 ~H-NMR (200 MHz, DMSO): 8 = 1.20 (t, 3H, CH2-CH3); 1.41 (s, 9H, tBu); 4.00-
4.28
(m, 4H, CH2-CH3 + Ns-CH2); 4.98 (s, 2H, N9-CH2); 8.09 (s, broad, 1 H, N6H);
8.15 +
8.21 (2 s, 2H, Cs-H + C8-H).
MS (ES+): m/e = 336.3 (M+H+, 100°~); 280.3 (60).
20 11 c) Ns-(2-Acetic acid)-N9-(tert-butyl 2-acetate)-adenine
249 mg (0.74 mmol) of Ns-(ethyl 2-acetate)-N9-(tert-butyl 2-acetate)-adenine
(Example 11 b) were dissolved in 6 ml of dioxane:waterariethylamine and
stirred at
RT for 4 days. The solvent was evaporated and the residue was chromatographed
25 through silica gel (DCM:MeOH 95:5 to 90:10). Yield: 36%.
MS (ES+): mle = 308.3 (M+H+, 100°~).
11 d) Ns-(2-(N-(2-tert-Butyloxycarbonylaminoethyl)acetamide))-N9-(tert-butyl 2-
acetate)-adenine
80 mg (0.26 mmol) of Ns-(2-acetic acid)-N9-(tert-butyl 2-acetate)-adenine
(Example
11 c), 42 mg (0.26 mmol) of 2-tert-butyloxycarbonylaminoethylamine were
dissolved
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66
in 5 ml of absol. DMF under argon and the mixture was treated at 0°C
with 85 mg
(0.26 mmol) of TOTU and 0.13 ml (0.78 mmol) of DIPEA and stirred at 0°C
for 10
min and at RT for 2.5 h. It was diluted to 100 ml using EA, then washed with
saturated potassium hydrogencarbonate solution, dried and concentrated. It was
chromatographed through silica gel (DCM:MeOH 98:2 to 90:10). Yield:
5°r6.
MS (ES+): mle = 450.3 (M+H+, 100°~).
11 e) N6-(2-(N-(2-Aminoethyl)acetamide))-N9-(2-acetic acid)-adenine
Synthesis analogously to Example 1 c from N6-(2-(N-(2-tert-
butyloxycarbonylaminoethyl)acetamide))-N9-(tert-butyl 2-acetate)-adenine
(Example
11 d).
Yield: 80°r6.
MS (ES+): mle = 293.1 (100°~; (M+H)+).
Example 12
Ns-(4-(2S-(Benzyloxycarbonylamino)butyric acid))-N9-(1-(3-guanidinylpropyl))-
adenine
12a) N9-(1-(3-(tert-Butyloxycarbonylamino)propyl))-6-chloropurine
154.6 mg (1 mmol) of 6-chloropurine were dissolved in 2.5 ml of absol. DMF and
treated with stirring with 331.7 mg (2.4 mmol) of K2C03 and 285.8 mg (1.2
mmol) of
tert-butyl N-(3-bromopropyl)carbamate. The mixture was stirred at RT for 11 h,
the
solvent was evaporated, the residue was taken up in EA and the solution was
washed twice with saturated NaHC03 solution, then with NaCI solution, dried,
filtered and concentrated. The residue was chromatographed through silica gel
(EA:n-heptane 8:2). Yield: 267 mg (86%).
~ H-NMR (200 MHz, DMSO): 8 = 1.37 (s, 9H, tBu); 2.00 (tt, 2H,
CH2-CH2-CH2); 2.95 (dt, 2H, CH2-NH); 4.30 (t, 2H, N9-CH2); 6.91 (t, broad, 1
H, NH);
8.70 + 8.78 (2 s, 2H, Cs-H + C8-H).
MS (ES+): mle = 312.2 (100%; (M+H)+); 256.1 (20).
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67
12b) N6-(4-(2S-(Benzyloxycarbonylamino)butyric acid))-N9-(1-(3-(tert-butyloxy-
carbonylamino)propyl))-adenine
370 mg (1.19 mmol) of N9-(1-(3-(tert-butyloxycarbonylamino)propyl))-6-
chloropurine
(Example 12a) were dissolved in 10 ml of absol. DMF and 5 ml of DIPEA. 449 mg
(1.8 mmol) of 2S-benzyloxycarbonylamino-4-aminobutyric acid were added at RT
and the mixture was stirred at 65°C for 50 h. The solvent was
evaporated and the
residue was partitioned between EA and saturated NaCI solution (20°~
KHS04). The
organic phase was washed with water, dried, ~Itered and concentrated. The
residue
was chromatographed through silica gel (EA:MeOH 8:2). Yield: 331 mg
(53°~).
~ H-NMR (200 MHz, DMSO): b = 1.39 (s, 9H, tBu); 1.73-2.21 (m, 2H, CH2-CH(NH-
Z)); 1.90 (m, 2H, CH2-CH2-CH2); 2.92 (dt, 2H, CH2-NHBoc); 3.15 (dt, 2H, N6H-
CH2);
3.88-4.10 (m, 1 H, CH-NHZ); 4.14 (t, 2H, N9-CH2); 5.03 (s, 2H, CH2-Ph); 6.91
(t,
broad, 1 H, NH-Boc); 7.37 (s, 5H, Ar-H); 7.55-7.81 (m, 2H, NH-Z + N6H-CH2);
8.13 +
8.19 (2 s, 2H, Cs-H + C8-H).
MS (ES+): mle = 528.2 (100%; (M+H)+).
12c) Ns-(4-(2S-(Benzyloxycarbonylamino)butyric acid))-N9-(1-(3-aminopropyl))-
adenine
30 mg (0.06 mmol) of Ns-(3-(2S-(benzyloxycarbonylamino)propionic acid))-N9-(1-
(3-
(tert-butyloxycarbonylamino)propyl))-adenine (Example 12b) were dissolved in 2
ml
of 90% strength trifluoroacetic acid, the solution was stirred at RT for 70
min and
concentrated, and the residue was stirred several times with ether. The
residue was
then dissolved in water, and the solution was freeze-dried. Yield:
100°~6.
MS (ES+): mle = 428.2 (100%; (M+H)+); 294.1 (90).
12d) N6-(4-(2S-(Benzyloxycarbonylamino)butyric acid))-N9-(1-(3-
guanidinylpropyl))-
adenine
Synthesis analogously to Example 1 d from N6-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(3-aminopropyl))-adenine
(Example
12c). Yield: 77%.
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68
MS (ES+): mle = 470.3 (25°~; (M+H)+); 336.2 (100).
Example 13
N6-(4-(2S-(Benzyloxycarbonylamino)butyric acid))-N9-(1-(3-(4,5-dihydro-1H-
imidazol-2-ylamino)propyl))-adenine
Synthesis analogously to Example 4 from Ns-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(3-aminopropyl))-adenine
(Example
12c). Yield: 63%.
MS (ES+): m/e = 496.3 (100°~; (M+H)+).
Example 14
N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(5-guanidinyl-
pentyl))-
adenine
14a) N9-(1-(5-(tert-Butyloxycarbonylamino)pentyl))-6-chloropurine
Synthesis analogously to Example 12a from 6-chloropurine and tert-butyl N-(5-
tosyloxypentyl)carbamate. Yield: 66°~.
~H-NMR (200 MHz, DMSO): b = 1.11-1.48 (m, 4H, 2 x CH2); 1.35 (s, 9H, tBu);
1.87
(tt, 2H, CH2); 2.97 (dt, 2H, CH2-NHBoc); 4.28 (t, 2H, N9-CH2); 6.72 (t, broad,
1 H,
NH); 8.71 + 8.78 (2 s, 2H, C6-H + C8-H).
MS (ES+): m/e = 340.2 (100%; (M+H)+); 284.1 (50).
14b) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(5-(tert-
butyloxycarbonylamino)pentyl))-adenine
Synthesis analogously to Example 12b from N9-(1-(5-(tert-
butyloxycarbonylamino)pentyl))-6-chloropurine and 2S-benzyloxycarbonylamino-
3-aminopropionic acid. Yield: 23%.
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~ H-NMR (200 MHz, DMSO): 8 = 1.10-1.49 (m, 4H, 2 x CH2; 1.36 (s, 9H, tBu);
1.62-
1.88 (m, 2H, CHZ); 2.87 (dt(2H, CH2-NHBoc); 3.68-4.98 (m, 5H, N9-CH2 + CH2-CH-
NHZ); 5.00 (s, 2H, CH2-Ph); 6.75 (t, broad, 1 H, NH); 8.02 + 8.20 (2 s, 2H, Cs-
H +
C8-H).
MS (FAB): mle = 542.3 (100%; (M+H)+).
14c) N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(5-amino-
pentyl))-
adenine
Synthesis analogously to Example 12c from N6-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(5-(tert-butyloxycarbonylamino)-
pentyl))-adenine (Example 14b). Yield: 100°~.
~ H-NMR (200 MHz, DMSO): S = 1.18-1.40 + 1.44-1.65 + 1.71-1.93 (2 m, 6H, 3 x
CH2); 2.77 (dt(2H, CH2-NHBoc); 3.64-4.35 (m, 5H, N9-CH2 + CH2CH-NHZ); 5.00 (s,
2H, CH2-Ph); 7.66 (m, 3H, NH3+); 8.20 + 8.24 (2 s, 2H, C6-H + C8-H).
MS (ES+): mle = 442.3 (40%; (M+H)+); 308.2 (100).
14d) N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(5-
guanidinylpentyl))-adenine
Synthesis analogously to Example 1 d from N6-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(5-aminopentyl))-adenine
(Example
14c). Yield: 90°~.
MS (ES+): mle = 484.3 (70°~; (M+H)+); 350.2 (60).
Example 15
N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(5-(4,5-dihydro-1 H-
imidazol-2-ylamino)pentyl))-adenine
Synthesis analogously to Example 4 from N6-(3-(2S-
(benzyloxycarbonylamino)propionic acid)-N9-(1-(5-aminopentyl))-adenine
(Example
CA 02225366 1997-12-19
14c). Yield: 75%.
MS (ES+): mle = 510.3 (40%; (M+H)+); 376.2 (100).
Example 16
5
N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(3-guanidinylpropyl))-
adenine
16a) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(3-(tert-
10 butyloxycarbonylamino)propyl))-adenine
Synthesis analogously to Example 12b from N9-(1-(3-(tert-
butyloxycarbonylamino)propyl))-6-chloropurine (Example 12a) and 2S-
benzyloxycarbonylamino-3-aminopropionic acid. Yield: 27%.
15 ~ H-NMR (200 MHz, DMSO): S = 1.37 (s, 9H, tBu); 1.90 (m, 2H, CH2-CH2-CH2);
2.92
(dt, 2H, CH2-NHBoc); 3.86 (m, broad, 2H, CH2-CH(NH-Z)); 4.13 (t, 2H, N9-CH2);
4.40 (m, 1 H, CH-NHZ); 5.01 (s, 2H, CH2-Ph); 6.92 (t, broad, 1 H, NH-Boc);
7.33 (s,
5H, Ar-H); 7.55-7.75 (m, 2H, NH-Z + NsH-CH2); 8.16 + 8.22 (2 s, 2H, Cs-H + C8-
H).
MS (ES+): mle = 514.3 (100°~; (M+H)+).
16b) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(3-amino-
propyl))-
adenine
Synthesis analogously to Example 12c from Ns-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(3-(tert-butyloxycarbonylamino)-
propyl))-adenine (Example 16a). Yield: 100%.
MS (ES+): mle = 414.2 (100%; (M+H)+); 280.2 (70).
16c) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(3-
guanidinylpropyl))-adenine
Synthesis analogously to Example 1 d from Ns-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(3-aminopropyl))-adenine
(Example
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71
16b). Yield: 98%.
MS (ES+): m/e = 456.3 (40%; (M+H)+); 322.2 (100).
Example 17
Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(4-guanidinyl-butyl))-
adenine
17a) N9-(1-(4-tert-Butyloxycarbonylamino)butyl))-6-chloropurine
Synthesis analogously to Example 12a from 6-chloropurine and tert-butyl N-(4-
tosyloxybutyl)carbamate. Yield: 66%.
~ H-NMR (200 MHz, DMSO): 8 = 1.30 (m, 2H, CH2); 1.35 (s, 9H, tBu); 1.86 (tt,
2H,
CH2); 2.93 (dt, 2H, CH2-NHBoc); 4.31 (t, 2H, N2-CH2); 6.79 (t, broad, 1 H,
NH); 8.72
+ 8.78 (2 s, 2H, Cs-H + C8-H).
MS (ES+): mle = 326.2 (80%; (M+H)+); 270.1 (100).
17b) N6-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(4-(tent-
butyloxy-
carbonylamino)butyl))-adenine
Synthesis analogously to Example 12b from N9-(1-(4-tert-
butyloxycarbonylamino)butyl))-6-chloropurine (Example 17a) and 2S-
benzyloxycarbonylamino-3-aminopropionic acid. Yield: 33%.
~ H-NMR (200 MHz, DMSO): 8 = 1.30 (m, 2H, CH2); 1.35 (s, 9H, tBu); 1.75 (m,
2H,
CH2); 2.91 (dt(2H, CH2-NHBoc); 3.71-4.34 (m,SH, CH2-CH(NH-Z) + N9-CH2); 5.01
(s, 2H, CH2-Ph); 6.89 (t, broad, 1 H, NH-Boc); 7.35 (s, 5H, Ar-H); 7.46-7.73
(m, 2H,
NH-Z + NsH-CH2); 8.10 (broad) + 8.20 (2 s, 2H, Cs-H + C8-H).
MS (FAB): mle = 528.4 (100%; (M+H)+).
17c) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(4-aminobutyl))-
adenine
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72
Synthesis analogously to Example 12c from N6-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(4-tert-butyloxycarbonyl-
aminobutyl))-adenine (Example 17b). Yield: 100°r6.
~ H-NMR (200 MHz, DMSO): 8 = 1.48 (m, 2H, CH2); 1.87 (m, 2H, CH2); 2.80 (dt,
2H,
CH2-NH2); 3.69-4.02 (m, 2H, CH2-CH(NH-Z)); 4.20 (t, 2H, N9-CH2); 4.36 (m,1H,
CH(NH-Z)); 5.01 (s, 2H, CH2-Ph); 7.33 (s, 5H, Ar-H); 7.64 (s, broad, 4H, NH3+
+
NsH-CH2); 8.10 (broad) + 8.20 (2 s, 2H, Cs-H + C8-H).
MS (ES+): mle = 428.3 (50%; (M+H)+); 294.2 (100).
17d) Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(4-
guanidinylbutyl))-adenine
Synthesis analogously to Example 1 d from Ns-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(4-aminobutyl))-adenine
(Example
17c). Yield: 78%.
MS (ES+): mle = 470.2 (50°~; (M+H)+); 336.2 (100).
Example 18
Ns-(3-(2S-(Benzyloxycarbonylamino)propionic acid))-N9-(1-(4-(4,5-dihydro-1 H-
imidazol-2-yl)amino)butyl))-adenine
Synthesis analogously to Example 4 from Ns-(3-(2S-
(benzyloxycarbonylamino)propionic acid))-N9-(1-(4-aminobutyl))-adenine
(Example
17c). Yield: 41 °~.
MS (ES+): mle = 496.3 (60%; (M+H)+); 362.2 (100).
Example 19
2S-Benzyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-yl-
carbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
19a) tert-Butyl 2S-benzyloxycarbonylamino-3-(6-(4-carboxypiperidin-1-yl)purin-
9-y1)-
propionate
CA 02225366 1997-12-19
73
260 mg (0.6 mmol) of tart-butyl 2S-benzyloxycarbonylamino-3-(6-chloropurin-9-
yl)-
propionate (Example 1 a), 116.3 mg (0.9 mmol) of piperidine-4-carboxylic acid
and
310 mg (2.4 mmol) of DIPEA in 4 ml of absol. DMF were stirred at 60°C
for 16 h. A
further 310 mg of DIPEA were then added and the mixture was again stirred at
60°C
for 24 h. The solvent was evaporated and the residue was partitioned between
EA
and water. The organic phase was washed again with KHS041K2S04 solution, then
with NaCI solution, dried, filtered and concentrated. The residue was
chromatographed through silica gel (EA). Yield: 219 mg (69°~).
MS (ES+): mle = 525.3 (100°~; (M+H)+).
19b) tert-Butyl 2S-benzyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-
2-
ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionate
126 mg (0.24 mmol) of tart-butyl 2S-benzyloxycarbonylamino-3-(6-(4-carboxy-
piperidin-1-yl)purin-9-yl)propionate (Example 19a), 39.3 mg (0.29 mmol) of 2-
amino-
1,4,5,6-tetrahydropyrimidine hydrochloride, 86.6 mg (0.264 mmol) of TOTU (O-
((ethoxycarbonyl)cyanomethylen-amino)-N,N,N',N'-tetramethyluronium
tetrafluoroborate (W. KtSnig et al., Proceedings of the 21 st European Peptide
Symposium 1990, E. Giralt, D. Andreu, Eds., ESCOM, Leiden, p. 143) and 124 mg
of DIPEA were added successively to 3 ml of absol. DMF. The solution was
stirred
at RT for 3 h, then a further 28 mg of DIPEA were added and the solution was
stirred at RT for 12 h. The reaction mixture was adjusted to pH 6 using
glacial acetic
acidltoluene (1:1 ), the reaction solution was concentrated, the residue was
partitioned between EA and saturated NaHC03 solution, and the organic phase
was
washed with NaCI, dried and concentrated. The residue was chromatographed
through silica gel (EA:MeOH:TEA 85:15:1.5). Yield: 70 mg.
MS (ES+): mle = 606.4 (60%; (M+H)+); 416.3 (40); 275.7 (100).
19c) 2S-Benzyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-y1-
carbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
80 mg of tent-butyl 2S-benzyloxycarbonylamino-3-(6-(4-(1,4,5,6-
tetrahydropyrimidin-
2-ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionate (Example 19b) were
dissolved in
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74
16 ml of precooled 95% strength trifluoroacetic acid and stirred first at
0°C for 30
min, then at RT for 30 min. The trifluoroacetic acid was removed in a rotary
evaporator, and the residue was coevaporated three times with toluene, stirred
in
ethanollether (1:2), washed with ether and dried in vacuo. Yield: 59 mg.
MS (ES+): m/e = 550.3 (60%; (M+H)+); 416.3 (100).
Example 20
2S-Benzyloxycarbonylamino-3-(1-(9-(2-guanidinoethyl)-9H-purin-6-y1)-1 H-
imidazol-
4-yl)propionic acid
20a) N9-(1-(2-(tert-Butyloxycarbonylamino)ethyl))-6-chloropurine
The synthesis was carried out analogously to Example 12a from 6-chloropurine
and
tart-butyl N-(2-tosyloxyethyl)carbamate. Yield: 36°~.
~ H-NMR (200 MHz, DMSO): b = 1.24 (s, 9H, tBu); 3.40 (dt, 2H, CH2-NHBoc); 4.35
(t,
2H, N9-CH2); 6.91 (t, broad, 1 H, NH); 8.60 + 8.78 (2 s, 2H, C6-H + C8-H).
MS (FAB): m/e = 298.2 (100°~; (M+H)+).
20b) 2S-Benzyloxycarbonylamino-3-(1-(9-(2-(tert-butyloxycarbonylamino)ethyl)-
9H-
purin-6-yl)-1 H-imidazol-4-yl)propionic acid
The synthesis was carried out analogously to Example 12b from N9-(1-(2-(tert-
butyloxycarbonylamino)ethyl))-chloropurine (Example 21 a) and Na Z-L-
histidine.
Yield: 33°~.
MS (ES+): mle = 551.3 (100°~; (M+H)+).
20c) 3-(1-(9-(2-Aminoethyl)-9H-purin-6-yl)-1 H-imidazol-4-yl)-2S-
benzyloxycarbonyl-
aminopropionic acid
The synthesis was carried out analogously to Example 12c from 2S-
benzyloxycarbonylamino-3-(1-(9-(2-(tart-butyloxycarbonylamino)ethyl)-9H-purin-
6-
y1)-1 H-imidazol-4-yl)propionic acid (Example 20b). Yield: 100%.
MS (ES+): mle = 451.3 (70°~; (M+H)+);
CA 02225366 1997-12-19
~ H-NMR (200 MHz, DMSO): b = 2.87-3.15 (m, 2H, Im-CH2); 3.38-3.51 (m, 2H, CH2-
NH2); 4.36 (m,1H, CH-NHZ); 4.60 (t, 2H, N9-CH2); 5.00 (s, 2H, CHZ-Ph); 7.28
(s, 5H,
aryl-H); 7.62 (d, 1 H, NH-Z); 8.23 + 9.05 (2 s, 2H, ImH); 8.71 + 8.88 (2 s,
2H, C6-H +
C8-H).
5
20d) 2S-Benzyloxycarbonylamino-3-(1-(9-(2-guanidinoethyl)-9H-purin-6-y1)-1 H-
imidazol-4-yl)propionic acid
The synthesis was carried out analogously to Example 1d from 3-(1-(9-(2-
10 aminoethyl)-9H-purin-6-yl)-1 H-imidazol-4-yl)-2S-benzyloxycarbonylamino-
propionic
acid (Example 20c). Yield: 38%.
MS (ES+): mle = 493.3 ((M+H) +);
Example 21
15 2R-Benzyloxycarbonylamino-3-(6-(N-(4-guanidinocyclohexyl)amino)purin-9-
yl)propionic acid
21 a) N9-(3-(tert-butyl 2R-(Benzyloxycarbonylamino)propionate))-6-chloropurine
20 The synthesis was carried out analogously to Example 1 a from 6-
chloropurine and
N-benzyloxycarbonyl-D-serine tert-butyl ester.
MS (FAB): mle = 432.2 (100°r6; (M+H)+); 376.1 (30).
21 b) tert-Butyl 2R-benzyloxycarbonylamino-3-(6-(N-(4-(tert-
25 butyloxycarbonylamino)cyclohexyl)amino)purin-9-yl)propionate
The synthesis was carried out analogously to Example 1b from 4-amino-1-(tert-
butyloxycarbonylamino)cyclohexane and N9-(3-(tert-butyl
2R-(benzyloxycarbonylamino)propionate))-6-chloropurine (Example 21 a). Yield:
30 55%.
MS (FAB): mle = 610.3 (100%; (M+H)+)
21 c) 3-(6-(N-(4-Aminocyclohexyl)amino)purin-9-yl)-2R-
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76
benzyloxycarbonylaminopropionic acid
The synthesis was carried out analogously to Example 1 c from tert-butyl 2R-
benzyloxycarbonylamino-3-(6-(N-(4-(tert-butyloxycarbonylamino)-
cyclohexyl)amino)purin-9-yl)propionate (Example 21 b). Yield: 100%.
MS (ES+): mle = 454.2 (50°~, (M+H)+)
21 d) 2R-Benzyloxycarbonylamino-3-(6-(N-(4-guanidinocyclohexyl)amino)-purin-9-
yl)propionic acid
The synthesis was carried out analogously to Example 1 d from 3-(6-(N-(4-
aminocyclohexyl)amino)purin-9-yl)-2R-benzyloxycarbonylaminopropionic acid
(Example 21 c). Yield: 80°~.
MS (ES+): mle = 496.3 (50%, (M+I-I)+).
Example 22
2R-Benzyloxycarbonylamino-3-(6-(N-(3-guanidinomethylbenzyl)amino)-purin-9-
yl)propionic acid
22a) tart-Butyl 2R-benzyloxycarbonylamino-3-(6-(N-(3-tert-
butyloxycarbonylaminomethylbenzyl)amino)purin-9-yl)propionate
The synthesis was carried out analogously to Example 1b from 3-aminomethyl-1-
(tert-butyloxycarbonylaminomethyl)benzene and N9-(3-(tert-butyl 2R-
(benzyloxycarbonylamino)propionate)-6-chloropurine (Example 21 a). Yield: 51
%.
MS (ES+): mle = 632.3 (100°~; (M+H)+).
22b) 3-(6-(N-(3-Aminomethylbenzyl)amino)purin-9-yl)-2R-
benzyloxycarbonylaminopropionic acid
The synthesis was carried out analogously to Example 1 c from tert-butyl 2R-
benzyloxycarbonylamino-3-(6-(N-(3-tert-butyloxycarbonyl-
aminomethylbenzyl)amino)purin-9-yl)propionate (Example 22a). Yield: 100%.
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77
MS (ES+): m/e = 476.2 ((M+H)+, 50%); 342.2 (70).
22c) 2R-Benzyloxycarbonylamino-3-(6-(N-(3-guanidinomethylbenzyl)amino)purin-9-
yl)propionic acid
The synthesis was carried out analogously to Example 1 d from 3-(6-(N-(3-
aminomethylbenzyl)amino)purin-9-yl)-2R-benzyloxycarbonylamino-propionic acid
(Example 22b). Yield: 30°~.
MS (ES+): m/e = 518.3 ( (M+H)+, 20°r6).
Example 23
3-(6-((4-(Benzimidazol-2-ylamino)butyl)amino)purin-9-yl)-2S-benzyloxycarbonyl-
aminopropionic acid
23a) 1-(4-tart-Butyloxycarbonylaminobutyl)-3-(2-nitrophenyl)thiourea
0.928 g (5.15 mmol) of 2-nitrophenyl isothiocyanate in 5 ml of absol. DMF was
added dropwise at 0°C to 0.97 g (5.15 mmol) of 4-(tent-
butyloxycarbonylaminobutyl)-
1-amine in 25 ml of absol. DMF. The solvent was distilled off and the residue
was
chromatographed through silica gel (EA:n-heptane 1:2 to 1:1 ). Yield: 1.8 g
(95°~).
MS (ES+): m/e = 369.2 ( M+H)+, 100%).
23b) 3-(2-Aminophenyl)-1-(4-tent-butyloxycarbonylaminobutyl)thiourea
1.78 g (4.8 mmol) of 1-(4-tent-butyloxycarbonylaminobutyl)-3-(2-nitro-
phenyl)thiourea (Example 23a) were dissolved in 120 ml of methanol and
hydrogenated (1 bar) over 1 g of PdIC at RT for 3 h. The catalyst was filtered
off, the
filtrate was concentrated and the residue was chromatographed through silica
gel
(EA:n-heptane 1:1 ). Yield: 1.4 g.
23c) 4-(Benzimidazol-2-ylamino)-1-(tert-butyloxycarbonylamino)butane
1.79 g (8.28 mmol) of yellow mercuric oxide and 27 mg of flowers of sulfur
were
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added to 1.4 g (4.14 mmol) of 3-(2-aminophenyl)-1-(4-tert-
butyloxycarbonylaminobutyl)thiourea (Example 23b) in 30 ml of ethanol and the
reaction mixture was heated at 50-55°C for 3 h. The solid was filtered
off with
suction and washed with ethanol. The filtrate was concentrated and the product
was
chromatographed through silica gel (DCM:methanol 9:5, then 9:1 ). Yield: 43%.
MS (ES+): mle = 305.2 ( (M+H)+, 100°~).
23d) 4-(Benzimidazol-2-ylamino)-1-aminobutane
198 mg (0.65 mmol) of 4-(benzimidazol-2-ylamino)-1-(tert-
butyloxycarbonylamino)butane (Example 23c) were dissolved in 20 ml of 95%
strength trifluoroacetic acid at 0°C and stirred at 0°C for 2 h,
then concentrated at
RT during the course of 30 min. The residue was coevaporated three times with
toluene, then stirred with ether and washed with pentane and dried in vacuo.
Yield:
100°r6.
MS (ES+): mle = 205.2 ( (M+H)+, 100°~).
23e) tert-Butyl 3-(6-((4-(benzimidazol-2-ylamino)butyl)amino)purin-9-yl)-
2S-benzyloxycarbonylaminopropionate
The synthesis was carried out analogously to Example 1 b from 4-(benzimidazol-
2-
ylamino)-1-aminobutane (Example 23d) and N9-(3-(tart-butyl 2S-
(benzyloxycarbonylamino)propionate))-chloropurine (Example 1 a). Yield: 32%.
MS (ES+): mle = 600.3 (100%; (M+H)+).
23f) 3-(6-((4-(Benzimidazol-2-ylamino)butyl)amino)purin-9-yl)-2S-
benzyloxycarbonylaminopropionic acid
The synthesis was carried out analogously to Example 1 c from tert-butyl 3-(6-
((4-
(benzimidazol-2-ylamino)butyl)amino)purin-9-yl)-2S-benzyloxy-
carbonylaminopropionate (Example 23e). Yield: 100%.
MS (ES+): mle = 544.2 ((M+H)+, 70%).
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79
Example 24
2S-Benzyloxycarbonylamino-3-(6-(4-((4,5-dihydro-1 H-imidazol-2-ylamino)-
methyl)piperidin-1-yl)purin-9-yl)propionic acid
24a) tent-Butyl 3-(6-(4-(Aminomethyl)piperidin-1-yl)purin-9-yl)-2S-
benzyloxycarbonylaminopropionate
The synthesis was carried out analogously to Example 1 b from 4-
(aminomethyl)piperidine and N9-(3-(tart-butyl 2S-(benzyloxycarbonylamino)-
propionate))-6-chloropurine (Example 1 a). Yield: 96.4°~.
MS (ES+): mle = 510.3 (100%; (M+H)+).
24b) 3-(6-(4-(Aminomethyl)piperidin-1-yl)purin-9-yl)-2S-
benzyloxycarbonylaminopropionic acid
The synthesis was carried out analogously to Example 1 c from tart-butyl 3-(6-
(4-
(aminomethyl)piperidin-1-yl)purin-9-yl)-2S-benzyloxycarbonylamino-propionate
(Example 24a). Yield: 100°~.
MS (ES+): mle = 454.3 ((M+H)+, 30°r6).
24c) 2S-Benzyloxycarbonylamino-3-(6-(4-((4,5-dihydro-1 H-imidazol-2-yl-
amino)methyl)piperidin-1-yl)purin-9-yl)propionic acid
The synthesis was carried out analogously to Example 4 from 3-(6-(4-
(aminomethyl)piperidin-1-yl)purin-9-yl)-2S-benzyloxycarbonylamino-propionic
acid
(Example 24b). Yield: 95%.
MS (ES+): mle = 522.3 ((M+H)+, 40%).
Example 25
2R-Benzyloxycarbonylamino-3-(6-(4-((4,5-dihydro-1 H-imidazol-2-ylamino)-
methyl)piperidin-1-yl)purin-9-yl)propionic acid
The synthesis was carried out analogously to Example 24 from N9-(3-(tert-butyl
2R-
CA 02225366 1997-12-19
(benzyloxycarbonylamino)propionate))-6-chloropurine (Example 21 a).
MS (ES+): mle = 522.3 ((M+H)+, 20°~).
Example 26
5 2S-Benzyloxycarbonylamino-3-(6-(4-(guanidinomethyl)piperidin-1-yl)purin-9-
yl)propionic acid
The synthesis was carried out analogously to Example 1 d from 3-(6-(4-
(aminomethyl)piperidin-1-yl)purin-9-yl)-2S-benzyloxycarbonylamino-propionic
acid
10 (Example 24b). Yield: 74°~.
MS (ES+): mle = 496.3 ((M+H)+, 40%).
Example 27
2S-Benzyloxycarbonylamino-3-(6-(3-(3-benzylureido)phenylsulfanyl)purin-9-
15 yl)propionic acid
27a) tart-Butyl 3-(6-(3-aminophenylsulfanyl)purin-9-yl)-2S-
benzyloxycarbonylaminopropionate
20 0.602 mmol of 3-mercaptoaniline were stirred in DMF and DIPEA for 12 h
together
with 0.602 mmol of N9-(3-(tart-butyl 2S-(benzyloxycarbonylamino)propionate))-6-
chloropurine (Example 1 a). The reaction solution was concentrated, the
residue was
partitioned between EA and saturated NaHC03 solution, the phases were
separated, the organic phase was washed with half-saturated NaHC03 solution
and
25 NaCI solution, dried and concentrated, and the product was chromatographed
through silica gel (EA:heptane 1:1). Yield: 190 mg.
MS (ES+): mle = 521.3 ( (M+H)+, 100°~).
27b) tart-Butyl 2S-benzyloxycarbonylamino-3-(6-(3-(3-benzylureido)-
30 phenylsulfanyl)purin-9-yl)propionate
46.1 mg of benzyl isocyanate in 1 ml of acetonitrile were added by means of a
syringe to 180 mg of tart-butyl 3-(6-(3-aminophenylsulfanyl)purin-9-yl)-2S-
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benzyloxycarbonylaminopropionate (Example 27a) in 3 ml of absol. acetonitrile.
The
mixture was stirred at RT for 48 h and concentrated, and the residue was
chromatographed through silica gel (DCM:EA 7:3 to 1:1). Yield: 205 mg.
MS (ES+): mle = 654.4 ( (M+H)+, 100%).
27c) 2S-Benzyloxycarbonylamino-3-(6-(3-(3-benzylureido)-phenylsulfanyl)purin-9-
yl)propionic acid
The synthesis was carried out analogously to Example 1 c from tert-butyl 2S-
benzyloxycarbonylamino-3-(6-(3-(3-benzylureido)phenylsulfanyl)purin-9-
yl)propionate (Example 27b). Yield: 100°~.
MS (ES+): mle = 598.4 ((M+H)+, 100°r6).
Example 28
2S-Neopentyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-yl-
carbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
28a) tart-Butyl 2S-amino-3-(6-(4-carboxypiperidin-1-yl)purin-9-yl)-propionate
1.7 g of tert-butyl 2S-benzyloxycarbonylamino-3-(6-(4-carboxypiperidin-1-
yl)purin-9-
yl)propionate (Example 19a) were dissolved in 200 ml of AcOH and hydrogenated
over PdIC at an H2 pressure of 1 atm. The catalyst was filtered off, the
solvent was
distilled off and the residue was lyophilized. Yield: 100%.
MS (ES+): mle = 391.3 ((M+H)+, 100°r6).
28b) tert-Butyl 2S-neopentyloxycarbonylamino-3-(6-(4-carboxypiperidin-1-
yl)purin-9-
yl)propionate
390 mg (1 mmol) of tert-butyl 2S-amino-3-(6-(4-carboxypiperidin-1-yl)purin-9-
yl)propionate (Example 20a) in 4 ml of DMF were treated at 0°C with 230
mg (1
mmol) of N-(neopentyloxycarbonyloxy)succinimide and 0.17 ml of DIPEA and,
after
slowly warming, stirred at RT for 12 h. The reaction mixture was concentrated
and
the residue was chromatographed (Lobar-C, DCM:MeOH:AcOH:H20 90:10:1:1).
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82
Yield: 540 mg.
MS (ES+): m/e = 505.4 ((M+H)+' 100%).
28c) tert-Butyl 2S-neopentyloxycarbonylamino-3-(6-(4-(1,4,5,6-
tetrahydropyrimidin-
2-ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionate
505 mg (1 mmol) of tert-butyl 2S-neopentyloxycarbonylamino-3-(6-(4-
carboxypiperidin-1-yl)purin-9-yl)propionate (Example 20b) were dissolved in 10
ml
of acetonitrile, treated with 250 mg of DCCI and 184 mg of pentafluorophenol
and
then stirred at RT for 30 minutes. The mixture was filtered, the mother liquor
was
concentrated, the residue was taken up in 5 ml of DMF, and the solution was
treated
with 200 mg of 2-amino-1,4,5,6-tetrahydropyrimidine and stirred at RT for 12
h. The
solvent was distilled off in vacuo and the residue was chromatographed (Lobar-
C,
DCM:MeOH:AcOH:H20 98:8:0.8:0.8). Yield: 270 mg.
MS (ES+): mle = 586.5 ((M+H)+, 100°r6).
28d) 2S-Neopentyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-
ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
The synthesis was carried out analogously to Example 19c from tert-butyl 2S-
neopentyloxycarbonylamino-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-ylcarbamoyl)-
piperidin-1-yl)purin-9-yl)propionate (Example 28c). Yield: 94%.
MS (ES+): mle = 530.4 ((M+H)+, 20%).
Example 29
2S-( 1-Adamantylmethyloxycarbonylamino)-3-(6-(4-( 1,4,5,6-tetrahydropyrimidin-
2-
ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
29a) tert-Butyl 2S-(1-adamantylmethyloxycarbonylamino)-3-(6-(4-carboxy-
piperidin-
1-yl)purin-9-yl)propionate
The synthesis was carried out analogously to Example 28b from N-(1-
adamantylmethyl-oxycarbonyloxy)succinimide and tert-butyl 2S-amino-3-(6-(4-
CA 02225366 1997-12-19
83
carboxypiperidin-1-yl)purin-9-yl)propionate (Example 28a). Yield: 85°~.
MS (ES+): mle = 583.4 ((M+H)+, 100%).
29b) tert-Butyl 2S-(1-adamantylmethyloxycarbonylamino}-3-(6-(4-(1,4,5,6-
tetrahydropyrimidin-2-ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionate
The synthesis was carried out analogously to Example 28c from tert-butyl 2S-(1-
adamantylmethyloxycarbonylamino)-3-(6-(4-carboxypiperidin-1-yl)purin-9-yl)-
propionate (Example 29a). Yield: 75°r6.
MS (ES+): mle = 664.5 ((M+H)+, 30°~).
29c) 2S-(1-Adamantylmethyloxycarbonylamino)-3-(6-(4-(1,4,5,6-
tetrahydropyrimidin-
2-ylcarbamoyl)piperidin-1-yl)purin-9-yl)propionic acid
The synthesis was carried out analogously to Example 19c from tart-butyl 2S-(1-
adamantylmethyloxycarbonylamino)-3-(6-(4-(1,4,5,6-tetrahydropyrimidin-2-yl-
carbamoyl)piperidin-1-yl}purin-9-yl)propionate (Example 29b). Yield:
100°~.
MS (ES+): mle = 608.4 ((M+H)+, 10%).
Pharmacological testing
The inhibition of binding of kistrin to human vitronectin receptor (VnR) is
described
below as a test method by which, for example, the antagonistic action of the
compounds according to the invention on the vitronectin receptor a~,f33 can be
determined (oc"f33 ELISA Test; the test method is abbreviated to "KNnR" in the
fisting of the test results).
Purification of kistrin
Kistrin is purified according to the methods of Dennis et al., as described in
Proc.
CA 02225366 2005-07-28
84
Natl. Ac:ad. Sci. USA 1989, 87, 2471-2475 and PROTEINS: Structure, Function
and
Genetics 1993, 15, 312-321.
Purification of human vitronectin receptor («,"f33)
Human vitronectin receptor is obtained from the human placenta according to
the
method of Pytela et al., Methods Enzymol. 1987, 144, 475. Human vitronectin
receptor a~,f33 can also be obtained from some cell lines (for example from
293 cells,
a human embryonic kidney cell line), which are co-transfected with DNA
sequences
for both subunits oc" and f33 of the vitronectin receptor. The subunits were
extracted
with octyl glycoside and then chromatographed through concanavalin A, heparin-
Sepharose and S-300.
Monoclonal antibodies
Murine monoclonal antibodies, specific for the f33 subunits of the vitronectin
receptor, are prepared according to the method of Newman et al., Blood, 1985,
227-
232, or Iby a similar process. The rabbit Fab 2 anti-mouse Fc conjugate to
horseradish peroxidase (anti-mouse Fc HRP) was obtained from Pel Freeze
(Cataloc,~ No. 715 305-1 ).
ELISA test
The abillity of substances to inhibit the binding of kistrin to the
vitronectin receptor
TM
can be determined using an ELISA test. For this purpose, Nunc 96-well
microtiter
plates are coated with a solution of kistrin (0.002 mg/ml) according to the
method of
Dennis et al., as described in PROTEINS: Structure, Function and Genetics
1993,
TM
15, 312-321. The plates are then washed twice with PBS/0.05% Tween-20 and
blocked by incubating (60 min) with bovine serum albumin (BSA, 0.5%, RIA grade
or
better) in Tris-HCI (50 mM), NaCI (100 mM), MgCl2 (1 mM), CaCl2 (1 mM), MnCl2
(1 mM), pH 7. Solutions of known inhibitors and of the test substances are
prepared
in concESntrations from 2 x 10-~2 to 2 x 10~ mol/l in assay buffer (BSA (0.5%,
RIA
CA 02225366 1997-12-19
grade or better) in Tris-HCI (50 mM), NaCI (100 mM), MgCl2 (1 mM), CaCl2 (1
mM),
MnCl2 (1 mM), pH 7). The blocked plates are emptied, and in each case 0.025 ml
of
this solution, which contains a defined concentration (2 x 10-2 to 2 x 10~
mol/l)
either of a known inhibitor or of a test substance, are added to each well.
0.025 ml
5 of a solution of the vitronectin receptor in the test buffer (0.03 mglml) is
pipetted into
each well of the plate and the plate is incubated at room temperature for 60-
180 min
on a shaker. In the meantime, a solution (6 mllplate) of a murine monoclonal
antibody specific for the f33 subunit of the vitronectin receptor is prepared
in the
assay buffer (0.0015 mglml). A second rabbit antibody (0.001 ml of stock
solutionl6
10 ml of the murine monoclonal anti-f33 antibody solution) which is an anti-
mouse Fc
HRP antibody conjugate is added to this solution, and this mixture of murine
anti-f33
antibody and rabbit anti-mouse Fc HRP antibody conjugate is incubated during
the
time of the receptor-inhibitor incubation. The test plates are washed four
times with
PBS solution which contains 0.05°~ Tween-20, and in each case 0.05
mllwell of the
15 antibody mixture is pipetted into each well of the plate and incubated for
60-180 min.
The plate is washed four times with PBS/0.05°~ Tween-20 and then
developed with
0.05 mllwell of a PBS solution which contains 0.67 mglml of o-phenylenediamine
and 0.012°~ of H202 . Alternatively to this, o-phenylenediamine can be
employed in
a buffer (pH 5) which contains Na3P04 and citric acid. The color development
is
20 stopped using 1 N H2S04 (0.05 mllwell). The absorption for each well is
measured
at 492-405 nm and the data are evaluated by standard methods.
The following test results were obtained:
25 Compound KIVnR KIVnR
of Example Inhibition at 10 NM (in °~) IC5o (NM)
1 75 1.1
2 80 0.7
30 3 77 2.2
4 93 0.15
12 86 0. 58
CA 02225366 1997-12-19
86
Compound KNnR KNnR
of Example Inhibition at 10 NM (in ICSO (NM)
%)
13 92 0.19
14 84 0.65
14c 22
92 0.21
16 85 0.54
16b 29
10 17 92 0.17
18 95 0. 075
19 97 0.004
23 93 0.16
24 95 0.052
15 25 89 0.345
26 91 0.36
