Note: Descriptions are shown in the official language in which they were submitted.
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
- 1 -
Modified oligosaccharides
The present invention relates to mimetics of sialyl-Lewis X and A, in which, in the natural
tetrasaccharide, the neuraminic acid residue is replaced by an S-configurated methyl sub-
stituted with one carboxyl residue and one other substituent and the natural N-acetyl group
in the N-acetylglucos~rnine monomer is replaced by a variety of different aliphatic and
aromatic substituents or the N-acetylglucosamine residue is replaced by a tetrahydropyran
derivative, to processes for the preparation of these compounds, to their use as a pharma-
ceutical and to pharmaceutical compositions co",plising them.
The complex process of inflammation, which takes place in several stages, is the body's
natural reaction to injuries in which, for example, there is also invasion by infectious agents
Under the influence of cytokines, the endothelium-which lines the blood vessels expresses
adhesion proteins on its surface. The P and E selectins bring about, by a protein-carbo-
hydrate interaction with glycolipids and glycoproteins on the leukocyte membrane, the so-
called "rolling" of leukocytes. The latter are slowed down by this process, and there is acti-
vation of certain proteins (integrins) on their surface which ensure firm adhesion of the
leukocytes to the endothelium. This is followed by migration of the leukocytes into the
damaged tissue.
There are many situations in which the recruitment of leukocytes by adhesion to the endo-
thelial cells is abnormal and in excess resulting in tissue damage instead of repair. This is
the case in disorders such as cardiogenic shock, myocardial infarct, thrombosis, rheuma-
tism, psoriasis, arthritis, dermatitis, acute respiratory di~l~ess syndrome, metastatic cancer
and transplantation.
One of the smallest natural carbohydrate epitopes as ligand for E selectin is sialyl-Lewis X
[neuraminic acid-a(2~3)-D-galactose-~(1 ~4)-L-(fucose-a(1 ~3))-N-acetyl-D-glucosamine
(sLe~)]. Although it has been considered to be potentially useful as an antiinflammatory
agent it can only be used as an injectable form as it is orally inactive and has a short half-
Iife in blood. Thus, there is a need for compounds which prevent the interaction between P
and F selectins and their receptors on the leukocyte membrane and which prevent the initial
cellular adhesion process.
CA 022608~4 1999-01-12
W 098/06730 PCT~EP97/04279 -2-
lt has now been found, surprisingly, that simultaneous replacement of the neuraminic acid
residue by an S-configurated methyt substituted with one carboxyl residue and one other
substituent and of the natural N-acetyl group in the GlcNAc monomer by a variety of diffe-
rent aliphatic and aromatic substituents or o~ the GlcNAc residue by a tetrahydropyran deri-
vative results in SLeX mimetics having interesting binding affinity properties.
According to the invention there is provided a compound of the formula I
OH
~ O
R'O ~ O
OH Z~o ~I)
o ~I~ OH
R ~ l~J OH
OH
wherein
R' is an S-configurated methyl substituted with a carboxy and one other substituent;
R2 is hydrogen, C,-C,2alkyl or C6aryl; where the alkyl and the aryl are unsubstituted or sub-
stituted by one or more substituents; and
Z is a group of the formula lla, llb or llc
'X R (lla) R~s~ (llb) 5~1~ (llc)
R
-
wherein
X is -C(O)-, -C(S)-, -S(0)2-, -C(O)Q- or -C(S)Q-, in which Q is NH, O, S, S-C1-C6alkylene,
NH-C,-C6alkylene or O-C1-C6alkylene;
RTl is C,-C,2alkyl, C2-C,2alkenyl, C,-Cl2alkoxy, C3-Cl2cycloalkyl, C3-Cl2cycloaikenyl,
C2-C"heterocycloalkyl, C2-C"heterocycloalkenyl, C6-C,0aryl, C6-C10aryloxy, C5-Cgheteroaryl,
Cs-Cgheteroaryloxy, C7-C"aralkyl, C7-C"aralkyioxy, C6-C10heteroaralkyl, C8-C,1aralkenyl or
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
- 3 -
C7-C10heteroaralkenyl, which are unsubstituted or substituted by one or more substituents;
and
RT2 is C3-Cl2cycloalkyl, C3-Cl2cycloalkenyl. C2-Cllheterocycloalkyl, C2-Cllheterocycloalkenyl,
C6-C1Oaryl, C6-C,Oaryloxy, Cs-c9heteroaryl~ Cs-c9heteroaryloxy~ C,-C"aralkyl, C,-C"aralkyl-
oxy, C6-C,Oheteroaralkyl, C8-C"aralkenyl or C,-ClOheteroaralkenyl, which are unsubstitut~sd
or substituted by one or more substituents;
R~s is NH2, primary amino, secondary amino or amido;
R5 i X~ RT1c c(O)NRT2cRT3c C(O~RT4c or C(o)oRT5c, wherein X' is C,-C4alkylene,
RT1C is hydrogen, halogen, C1-C12alkyl, C,-C,lheteroalkyl, C3-C,2alkenyl, C3-C~2cycloalkyl,
C3-C,2cycloalkenyl, C2-C1,heterocycloalkyl, C2-C"heterocycloalkenyl, C6-, C,O- or C,4aryl,
C2-Cgheteroaryl, C7-C1,aralkyl, C6-C,Oheteroaralkyl, Cg-C"aralkenyl, C8-C,Oheteroaralkenyl,
oRT6c oc(o)RT4c SRT4C S02RT9C or S03R
each of RT2C, RT3C and RT4C is independently hydrogen, C,-C12alkyl, C,-C"heteroalkyl,
C3-C,2alkenyl, C3-C,2cycloalkyl, C3-C,2cycloalkenyl, C2-C"heterocycloalkyl, C2-C"hetero-
cycloalkenyl, C6-, C,O- or C14aryl, C2-Cgheteroaryl, C,-C,laralkyl, C6-ClOheteroaralkyl,
Cg-C,,aralkenyl or C~-C,Oheteroaralkenyl;
each of RTsC~ RT7C and RT9C is independently hydrogen, Myl C,-C,2alkyl, C,-C"heteroalkyl,
C3-Cl2alkenyl, C3-C,2cycloalkyl, C3-C,2cycloalkenyl, C2-C"heterocycloalkyl, C2-C"hetero-
cycloalkenyl, C6-, C,O- or C14aryl, C2-Cgheteroaryl, C,-C"aralkyl, C6-C10heteroaralkyl,
Cg-C,,aralkenyl or C8-C,Oheteroaralkenyl;
RT6C is hydrogen, C,-C,2alkyl, C,-C1,heteroalkyl, C3-C,2alkenyl, C3-C,2cycloalkyl,
C3-C,2cycloalkenyl, C2-C,1heterocycloalkyl, C2-C11heterocycloalkenyl, C6-, C10- or C14aryl,
C2-Cgheteroaryl, C7-C"aralkyl, C6-C,Oheteroaralkyl, Cg-C,,aralkenyl, C8-C,Oheteroaralkenyl,
SO RT5c pO RT7cRT8C c(o)oRT9c C(s)NRT2cRT3c orC(O)NRT2cRT3c; and
RT9C is C,-C,2alkyl, C,-C"heteroalkyl, C3-C,2alkenyl, C3-C,2cycloalkyl, C3-Cl2cycloalkenyl,
C2-C"heterocycloalkyl, C2-Cl,heterocycloalkenyl, C6-, Clo- or C,4aryl, C2-Cgheteroaryl,
C7-C11aralkyl, C6-C,Oheteroaralkyl, Cg-C,,aralkenyl or C8-ClOheteroaralkenyl;
wherein the s~hstitl~ent is selected from the group consisting of OH, halogen, NH2, C(O)Rs2,
C(O)ORs', oC(o)Rs4, nitro, cyano, SO3H, OSO3H, S03Myl OSO3My~ NR2~SO3My, C,-Cl2alkyl,
C2-C12alkenyl, C,-Cl2alkoxy, C3-C,2cycloalkyl, C3-Cl2cycloalkenyl, C2-Cllheterocycloalkyl,
C2-CI,heterocycloalkenyl, C6-C,Oaryl, C6-ClOaryloxy, Cs-c9heteroaryl~ Cs-c9heteroaryloxy~
C7-C"aralkyl, C7-C"aralkyloxy, C6-C,Oheteroaralkyl, C8-C"aralkenyl, C7-ClOheteroaralkenyl,
primary amino, secondary amino, sulfonyl, sulfonamido, carbamido, carbamate, sulfonhydr-
azido, carbhydrazido, carbohydroxamic acid and aminocarbonylamido, where Rs' is hydro-
.
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97104279
-4-
gen, My~ C,-C,2alkyl, C2-C12alkenyl, C3-C,2cycloalkyl, C2-C"heterocycloalkyl, C6-C10aryl,
C5-Cgheteroaryl, C7-C"aralkyl or C6-C10heteroaralkyl, Rs4 is hydrogen, C1-C12alkyl,
C2-C12alkenyl, C3-C12cycloalkyl, C2-C11heterocycloalkyl, C~-C,Oaryl, C5-Cgheteroaryl,
C7-C"aralkyl or C6-C,0heteroaralkyl, and Rs2 and R20 are hydrogen, C1-C12alkyl,
C2-C,2alkenyl, C3-Cl2cycloalkyl, C3-C,2cycloalkenyl, C2-C,1heterocycloalkyl, C2-Cl1-hetero-
cycloalkenyl, C6-C10aryl, C5-Cgheteroaryl, C7-C11aralkyl, C6-C,0heteroaralkyl, C8-C11-aralkenyl
or C7-C,Oheteroaralkenyl, and alkyl, alkenyl, alkoxy, cycloalkyl, cycloalkenyl, heterocyclo-
alkyl, heterocycloalkenyl, aryl, aryloxy, heteroaryl, heteroaryloxy, aralkyl, aralkyloxy, hetero-
aralkyl, aralkenyl and heteroaralkenyl in turn are unsubstituted or substituted by one of the
abovementioned substituents; and y is 1 and M is a monovalent metal or y is 1/2 and M is a
divalent metal; and a derivative thereof wherein at least one OH is substituted with SO~RTsC,
PO3R R , C(O)R , C(O)ORT9C, C(S)NRT2CRT3c C(o)NRT2CRT3c C C Ik I
kenyl, C3-C,2cycloalkyl, C3-C,2cycloalkenyl, C,-C"heteroalkyl, C2-C"heterocycloalkyl,
C2-C"heterocycloalkenyl, C6-, C10- or C14aryl, C2-C9heteroaryl, C7-C"aralkyl, C6-C,0hetero-
aralkyl, Cg-C,,aralkenyl or C8-C,0heteroaralkenyl; in free forrrl or in salt form.
Preferably, Z is bound to the galactose moiety via the carbon atom 4 in case of formula lla
and via the carbon atom 3 in case of formulae llb and llc.
M is preferably an alkali metal (for example lithium, sodium, potassium, rubidium and
caesium), an alkaline earth metal (for example magnesium, calcium and strontium) or
manganese, iron, zinc or silver.
Halogen is fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine,
especially fluorine or chlorine.
Alkyl may be linear or branched, preferably branched once or twice in the a position.
Examples of alkyl include e.g. methyl, ethyl and the isomers of propyl, butyl, pentyl, hexyl,
heptyl, octyl, nonyl, decyl, undecyl and dodecyl, preferably methyl, ethyl, n- and i-propyl, n-,
i- and t-butyl. Examples of alkenyl are allyl, but-1-en-3-yl or -4-yl, pent-3- or 4-en-1-yl or
-2-yl, hex-3- or -4- or -5-en-1-yl or-2-yl and (C,-C4alkyl)CH=CH-CH2-. Examples of alkylene
are ethylene, 1 ,2-propylene, 1,2- or 2,3-butylene, 1,2- or 2,3-pentylene, 1,2-, 2,3- or
3,4-hexylene.
CA 022608~4 1999-01-12
W 098106730 PCT~EP97/04279
-5-
Cycloalkyl and cycloalkenyl may contain 5 to 8, preferably 5 or 6 carbon atoms. Examples
of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl,
~ preferably cyclohexyl. Examples of cycloalkenyl are cyclopropenyl, cyclobutenyl, cyclopent-
enyl, cyclohexenyl, cycloheptenyl and cyclooctenyl, preferably cyclohexenyl. Examples of
cycloalkylene are 1,2-cyclopropylene, 1,2-cyclobutylene, 1,2-cyclopentylene, 1,2-cyclo-
hexylene, 1,2-cycloheptylene and 1,2-cyclooctylene. Examples of heterocycloalkylene are
pyrrolidinylene, piperidinylene, tetrahydrofuranylene, di- and tetrahydropyranylene.
Examples of heterocycloalkyl are derived from pyrrolidine, imidazolidine, oxazolidine,
pyrazolidine, piperidine, piperazine and morpholine. Examples of heterocycloalkenyl are
derived from 2- and 3-pyrroline, oxazoline, 2- and 4-imidazoline and 2- and 3-pyrazoline.
Aryl or heteroaryl is a five- or six-membered ring or a bicycle consisting of two condensed
six- or five-membered rings or one six-membered and one five-membered rin~, and in the
case of heteroaryl one or more C atoms may be replaced, independently of one another, by
an atom selected from oxygen, nitrogen and sulfur. Examples are derived from benzene,
naphthalene, indene, furan, pyrrole, pyrazole, i~, d~ole, isoxazole, oxazole, furazan, thia-
diazole, thiophene, thiazole, ox~di~ole, triazole, indole, indazole, purine, benzimidazole,
benzoxazole, benzothiazole, pyran, pyridine, pyridazine, triazine, pyrimidine, pyrazine, iso-
quinoline, cinnoline, phthalazine, quinoline, quinazoline, pterdine, benzotriazine or quinoxa-
line. Aryl is preferably naphthyl and phenyl, particularly phenyl. Heteroaryl is preferably
furanyl, pyridinyl and pyrimidinyl.
Aralkyl preferably has 7 to 12 C atoms and may be phenyl-CnH2n- with n equal to a number
from 1 to 6. Examples are benzyl, phenylethyl or phenylpropyl. Benzyl and 2-phenylethyl
are preferred. Aralkenyl is preferably unsubstituted cinnamyl or cinnamyl ring-substituted by
a substituent selected from the group consisting of OH, halogen, COOH, C(O)OMy~
C,-C,2alkyl, C,-C6alkoxy, C6-C,0aryl, SO3My~ OSO3My~ NR2~SO3My in which R20 is as defined
above. Heteroaralkyl and heteroaralkenyl are preferably C4-C5heteroarylmethyl and
C4-Csheteroarylethenyl with one or two hetero atoms from the group of O and N, and the
heteroaryl may comprise the abovementioned heteroaryl residues.
Alkoxy may be linear or branched, preferably branched once or twice in the a position.
Examples of alkoxy include e.g. methoxy, ethoxy and the isomers of propoxy, butoxy, pent-
oxy, hexoxy, heptoxy, octoxy, nonoxy, decoxy, undecoxy and dodecoxy, preferred are
CA 022608~4 1999-01-12
W O 98/06730 PCT/EP97/04279
-6-
methoxy and ethoxy. Examples of aryloxy and aralkoxy are phenoxy and benzyloxy.
Heteroaryloxy is preferably furanyloxy, pyridinyloxy and py,il";di, Iyloxy.
The primary amino preferably contains 1 to 12, particularly preferably 1 to 6,C atoms, and
may be e.g. methyl-, ethyl-, hydroxyethyl-, n- or i-propyl-, n-, i- or t-butyl-, pentyl-, hexyl-,
cyclopentyl-, cyclohexyl-, phenyl-, methylphenyl-, benzyl- and methylbenzylamino. The
secondary amino preferably contains 2 to 14, particularly preferably 2 to 8, C atoms, and
may be e.g. dimethyl-, diethyl-, methylethyl-, di-n-propyl-, di-i-propyl-, di-n-butyl-, diphenyl-,
dibenzylamino, morpholino, piperidino and pyrrolidino.
Primary amino and secondary amino preferably correspond to R8 R9N in which each R8 and
R9 is independently hydrogen, OH, SO3My~ OSO3My~ C,-C,2alkyl, C3-C,2cycloalkyl,
C2-Cl1heterocycloalkyl, C6-C,0aryl, Cs-c9heteroaryl~ C,-C"aralkyl, C6-C,0heteroaralkyl,
C8-C,6aralkenyl with C2-C6alkenylene and C6-C10aryl, or di-C6-C,0aryl-C,-C6-alkyl, which are
unsubstituted or substituted by one or more of the above substituents; or R8 and R9 to-
gether are tetramethylene, pentamethylene, -(CHz)20(CH2)2-, -(CH2)2S(CH2)2- or
-(CH2)2NR7(CH2)2-, and R' is H, C,-C6alkyl, C7-C,1aralkyl, C(O)Rs2 or sulfonyl.
Carbamido, carbamate, carbhydrazido, sulfonamido, sulfonhydrazido and aminocarbonyl-
amido preferably correspond to a group R3C(o)(NH)pN(R9)-, -C(O)(NH)pNR8R9,
R80C(O)(NH)pN(R9)-, R8R40NC(o)(NH)pN(R9)-,-oC(o)(NH)pNR8R9~-N(R4o)c(o)(NH)pNR8R
R8S(0)2(NH)pN(R9)-; -S(0)2(NH)pNR8R9; R8R4~NS(o)2N(R9)-, -NR4~S(o)2NR8R9 or
-N(R40)C(o)C(o)NR8R9, in which each of R8, R9 and R40 is independently hydrogen, OH, C,-
C,2alkyl, C,-C,2alkenyl, C3-C,2cycloalkyl, C3-C,2cycloalkenyl, C2-Cl,heterocycloalkyl, C2-
C1,heterocycloalkenyl, C6-C,0aryl, Cs-c9heteroaryl~ C7-C,6aralkyl, C8-C,6aralkenyl with C2-
C6alkenylene and C6-C,0aryl, C6-C~sheteroaralkyl~ C6-C~sheteroaralkenyl~ or di-C6-C10aryl-C1-
C6-alkyl; or R8 and R9 or R8 and R40 in the case of -NR8R9 or R8R40N- together are
tetramethylene, pentamethylene, -(CH2)2-O-(CH2)2-, -(CH2)2-S-(CH2)2- or -(CH2)2-NR -(CH2)2-
, and R' is H, C,-C6alkyl, C7-C"aralkyl, C(O)Rs2 or sulfonyl.
The sulfonyl substituent corresponds, for example, to the formula R1~-S02- in which R10 is
C,-C,2alkyl, C3-C,2cycloalkyl, C2-C"heterocycloalkyl, C6-C1Oaryl, C5-Cgheteroaryl, C7-C11ar-
alkyl or C6-C,Oheteroaralkyl.
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97/04279
-7-
The other substituent in R' has preferably 1 to 20, more preferably 1 to 16, particularly pre-
ferably 1 to 12, and especially preferably 1 to 8 C atoms. The other s~ ~bstih ~ent is preferably
selected from the group consi~ g of unsu~stituted and suhstitl~ted C,-Cl2alkyl,
C2-C,2alkenyl, C3-Ct2cycloalkyl, C3-Cl2cycloalkenyl, Crc"heterocycloalkyl~ C2-C"hetero-
cycloalkenyl, C6-C,0aryl, C5-Cgheteroaryl, C7-Cl,aralkyl, C6-Cl0heteroaralkyl, C8-Cl,aralkenyl
and C7-C,Oheteroaralkenyl. The other substituent is particularly substituted methyl, or 2-sub-
stituted ethyl or unsubstituted cyclohexyl. Examples of suitable s~hstihlents are the substi-
tuents mentioned above in the definition of R2, especially OH, halogen (F, Cl or Br), carb-
oxyl, -SO3H, C(O)OMy~ SO3My~ OSO3Myt NR2~SO3My in which R20 is as defined above, or
C,-C,2alkyl, C,-C,2alkoxy, nitro, -NH2, primary amino with 1 to 20 C atoms, secondary amino
with 2 to 30 C atoms, cyano, C3-C8cycloalkyl, C3-C6heterocycloalkyl, C6-Cl0aryl,C3-Cgheteroaryl, C,-C,6heteroaralkyl, where the hetero atoms are selected from the group of
O, S and N atoms, and carbamide, carbamate, carbhydrazide, sulfonamide, sulfonhydr-
azide or aminocarbonylamide, whose N atoms are unsùbstituted or substituted by a hydro-
carbon group or hydroxy-hydrocarbon group with 1 to 20 C atoms. The hydrocarbon groups
and heterohydrocarbon groups in turn are unsubstituted or substituted, for example with
C1-C6alkyl, Cl-C6alkoxy, carboxyl, halogen (F, Cl or Br), -OH, -CN or -NO2.
In a particular embodiment of the compounds of the formula 1, R' corresponds to a group of
the formula ll,
CoOR3
H /,~ (Il),
R4
in which R3 is hydrogen or My; and R4 is Cl-C12alkyl, C2-C,2alkenyl, C3-C,2cycloalkyl,
C3-C,2cycloalkenyl, C2-Cllheterocycloalkyl, C2-C"heterocycloalkenyl, C6-C,Oaryl,Cs-Cgheteroaryl, C7-C11aralkyl, C6-C10heteroaralkyl, C8-C1,aralkenyl or C7-C,Oheteroar-
alkenyl, which are unsubstituted or substituted by one or more substituents selected from
the abovementioned group of substituents.
Preferred compounds of the formula I are those in which R1 corresponds to a group of the
formula ll in which R3 is hydrogen or My and R4 is
(a) unsubstituted C1-C,2alkyl; C,-C,2alkyl which is substituted by one or more substituents
selected from the group consisting of -NH2, primary amino, secondary amino, C,-C,2sul-
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97/04279
-8-
fonyl, carbamide, carbamate, carbhydrazide, sulfonamide, sulfonhydrazide, amino-carbonylamido, C3-C,2cycloalkyl, C,-C6alkoxy, phenyloxy and benzyloxy; unsubstituted
C3-Cl2cycloalkyl; C3-Cl2cycloalkyl which is s~ stituted by one or more substituents selected
from the group consisting of Cl-C6alkyl, Cl-C6alkoxy, C,-C12sulfonyl, phenyloxy and benzyl-
oxy; C6-C10aryl; C3-Cgheteroaryl with 1 or 2 hetero atoms sel~ctecl from oxygen and nitro-
gen; C7-C16aralkyl with C,-C6alkyl and C6-C10aryl; C4-C16heteroaralkyl with Cl-C6alkyl and C3-
C,Oheteroaryl with 1 or 2 hetero atoms selected from oxygen and nitrogen and a total of 3 to
5 carbon atoms; or such C6-C,0aryl, C3-Cgheteroaryl, Crc~6aralkyl and C3-C16heteroaralkyl
which are substituted by one or more substituents selected from the group consisting of
OH, halogen, C1-C12sulfonyl, carboxyl, C(O)OMy~ C1-C12alkyl, C1-C6alkoxy, C6-C10aryl,
SO3My~ OSO3My~ NR2~SO3My in which R20, y and M are as defined above; or
(b) C,-C12alkyl, C3-C,2cycloalkyl or C7-C11aralkyl, in particular CH2-C6Hs and (CH2)2-C6H5,
which are unsubstituted or substituted by one or more substituents selected from the
abovementioned group of substituents.
More preferred are those compounds in which the substituent for R4 is selected from the
group consisting of NH2, C3-C,2cycloalkyl, primary amino, secondary amino, sulfonamido,
carbamido and aminocarbonylamido. Particularly preferred substituents for C1-C12alkyl are
NH2, cyclohexyl, C6-C10aryl, R8R9N-, R8C(O)N(R9)-, R6S(0)2N(R9)-, R8NHC(O)NR9- and
NR9C(o)NHR8 in which R8, R9, R~ and R9 are as defined above.
Particulariy preferred compounds within this group are those in which R4 is R'4, R'4 being
CH2-C6Hs, tCH2)2-C6H~, cyclohexyl, methyl, ethyl or isopropyl which are unsubstituted or
substituted by one or more substituents selected from the group consisting of NH2, cyclo-
hexyl, C6-C,0aryl, R8C(O)N(R9)-, R8S(0)2N(R9)-, R8NHC(O)NR9-, NR9C(o)NHRa and R3R9N-,
in which each R8, R9, R3 and R9 is independently hydrogen, C1-Cl2alkyl, C3-C12cycloalkyl,
C6-C,0aryl or C7-C,1aralkyl, which are unsubstituted or substituted by one or more substitu-
ents selected from the group consisting of OH, halogen, C(O)OMy~ nitro, cyano, SO3My~
OSO3My~ NHSO3Myl C1-C12alkyl, C1-Clzalkoxy and C6-C,0aryl, where y and M are as defined
above. Particularly preferred compounds are those in which each R8, R9, R3 and R9 is inde-
pendently hydrogen, C,-C,2alkyl, cyclohexyl, phenyl, naphthyl or C,-C,1aralkyl, which are
unsubstituted or substituted by one or more substituents selected from the group consisli"g
of OH, F, Cl, C(O)ONa, nitro, cyano, SO3Na, Cl-C6alkyl, methoxy and phenyl.
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
9.
In a preferred group of compounds of the formuia 1, R1 corresponds to formula 11, in which
R4 is R"4, R"4 being C6Hl1, CH(CH3)2, CH2-phenyl, (CH2)2-phenyl, CH2NHC(O)-phenyl,
CH2NHC(O)(CH2)3-phenyl, CH2N~IC(O)(CH2)30H, CH2NHC(O)CF3, CH2NHC(O)C6Hl1,
CH2NHC(O)Cl,H23, CH2NHc(o)cH(c6H5)2~ CH2HNc(o)NHc6H5~ C~12NHC(O)C2H4CO2Na,
CH2NHC(O)C6[(1,3,4,5)0H]4H7, CH2NHC(O)C6H4-p-SO3Na, CH2NHC(O)C6H4CI, CH2C6H11,
(CH2)2C6Hl1, CH2NH2~ CH2NHc(o)c6H4No2~ CH2NHc(o)c6H4ocH3~ CH2NHc(o)c1oH7
CH2NHC(O)C6H4(3,4)CI2, CH2NHc(o)c6H4cH3~ CH2NHc(o)c6H4c6Hs~ CH2NHc(o)c6H4cN~
CH2NHC(O)C6H4COONa, CH2NHC(O)(CHOH)2COONa, CH2N[CH2CH(CH3)2][C(O)-phenyl],
CH2N(CH2CH=CH-phenyl)[C(O)-phenyl]. CH2N[c(o)c6H5]cH2c6Hs~ Ct~2NHCH2-phenyl,
CH2N[C(O)C6H5](CH2)3C6H5, CH2NHCH2CH=CH-phenyl, CH2NHCH2CH(CH3)2,
CH2N(CH2-phenyl)2, CH2N[CH2CH(CH3)2]2, CH2NHSO2-p-nitrophenyl, CH2NHso2-p-t
CH2NHSO2CF3, CH2NHC(O)NHC6H5 or CH2N[SO2-p-nitrophenyl][CH2CH(CH3)2]2.
A preferred group of compounds of the formula I are those in which R2 is hydrogen, unsub-
stituted or substituted C,-C6alkyl, preferably C1-C4alkyl, especially methyl or ethyl, wherein
the substituent is selected from C(O)OH, -C(O)ONa, -C(O)OK, -OH, -C(O)-NR~ R9 and
-SO2-NR8 R9, in which R8 is H, C,-C4alkyl, C2-C4hydroxyalkyl, phenyl or benzyl, and R9 in-
dependently has the meaning of R8, or R8 and R9 are together tetramethylene, pentameth-
ylene or -CH2CH2-O-CH2CH2-. Particularly preferred compounds are those in which R2 is
hydrogen, methyl, ethyl, HO(O)CCH2CH2-, NaOC(O)CH2CH2- or R8 R9 NC(O)CH2CH2-, and
R~ and R9 are, independently of one another, ~, C1-C6alkyl, C2-C4hydroxyalkyl, phenyl,
benzyl or, together, morpholino.
A first preferred embodiment of the invention comprises the compounds of formula IA
oRTl
o~ X,RT''
~ O
~~ rOH ~OH (IA)
HO~/ R ' - OH
. ORl OH
in which X Rt R2 RTl and RT2 have the above meanings
CA 022608~4 l999-0l-l2
W O 98/06730 PCT~EP97/04279
-10-
Q in X is preferably NH, O or S. X is preferably -C(O)-, -C(S)-, -C(O)O- or-C(S)O-, more
preferably -C(O)- or-C(O)O-.
A preferred embodiment of the invention are those compounds of the formula IA wherein
RT' is C1-C,2alkyl, which is unsubstituted or s~hstitllted by one or more substituents selec-
ted from the group consisting of OH, halogen, C(O)ORs', oC(o)Rs4, C(O)Rs2, nitro, NH2,
cyano, SO3My~ OSO3My~ NR2~S03My1 where Rs', Rs4, Rs2, R20, y and M are as defined above.
A more preferred embodiment of the invention are those compounds of the formula IA
wherein RT' is C,-C,2alkyl, which is unsubstituted or substituted by one or more, preferably
one C(O)ORs', where Rs' is as defined above. Most preferably RT1 is C,-C,2alkyl, which is
substituted by C(O)OC,-C,2alkyl or C(O)ONa. A specially preferred meaning of RT' is
(CH2)QC(O)OCH3 or (CH2)aC(O)ONa.
A preferred embodiment of the invention are those compounds of the formula IA wherein
RT2 is C3-C,2cycloalkyl, C2-C,lheterocycloalkyl, C6-C,Oaryl or C5-C9heteroary1, which are
unsubslituted or substituted by one or more substituents selected from the group consisting
of OH, halogen, nitro, NH2, cyano, C,-C,2alkyl, C2-C,2alkenyl, C,-C12alkoxy, C3-C,2cycloalkyl,
C3-Cl2cycloalkenyl, C2-C,1heterocycloalkyl, C2-C1,heterocycloalkenyl, C6-C,Oaryl, C6-C10aryl-
oxy, Cs-Cgheteroaryl, C5-Cgheteroaryloxy, C7-Cl,aralkyl, C7-C,1aralkyloxy, C6-ClOheteroar-
alkyl, C~-C,1aralkenyl, C7-C,Oheteroaralkenyl. More preferred are those compounds of the
formula IA wherein RT2 is C3-C12cycloalkyl, C2-C11heterocycloalkyl, C6-Cl0aryl or
Cs-Cgheteroaryl, which are unsubstituted or substituted by one or more substituents
selected from the group consisting of OH, C1-Cl2alkyl, C6-C,0aryl or Cs-c9heteroaryl. Most
preferred meanings of RT2 are -3,5-(OH)2C6H3, -3,4-(OH)2C6H3, -3,4-(OCH3)2C6H3,
-2-(OH)C6H4 and thyminyl, especially preferred are -3,4-(OH)2C6H3 and -3,4-(OCH3)2C6H3.
A particularly preferred embodiment of the invention comprises compounds of the formula
laA
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
ORT1
RT2
~, O
HO;~ O~
~, O OH
H CoOR3
wherein X, R3, R4, RT' and RT2 are as defined above.
Preferred compounds of the formula laA are those in which X is -C(O)-, -C(S)-, -S(0)2-,
-C(O)Q- or -C(S)Q-, in which Q is NH, O or S; R3 is hydrogen or My; R4 is C7-Cl,aralkyl,
C3-C,2cycloalkyl or C1-C,2alkyl, which is unsubstituted or substituted by one or more substi-
tuents selected from the group consisting of NH2, C3-C12cycloalkyl, primary amino, secon-
dary amino, sulfonamido, carbamido and aminocarbonylamido; RT' is C,-C12alkyl, which is
unsubstituted or substituted by one or more substituents selected from the group consisting
of OH, halogen, C(O)ORsl, oC(o)Rs4, C(O)Rs2, nitro, NH2, cyano, SO3Myl OSO3My~
NR2~SO3Myl where Rs', Rs4, Rs2, R20, y and M are as defined above; and RTZ is C3-C,2cyclo-
alkyl, C2-C1,heterocycloalkyl, C6-C,0aryl or Cs-c9heteroaryll which are unsubstituted or sub-
stituted by one or more substituents selected from the group consisting of OH, halogen,
nitro, NH2, cyano, C,-C,2alkyl, C2-C,2alkenyl, C,-C,2alkoxy, C3-C,2cycloalkyl, C3-C,2cyclo-
alkenyl, C2-C"heterocycloalkyl, C2-C"heterocycloalkenyl, C6-C,0aryl, C6-C10aryloxy,
Cs-Cgheteroaryl, Cs-Cgheteroaryloxy, Crc"aralkyl~ Crcl~aralkyloxy~ C6-C,Oheteroaralkyl,
C8-Cl1aralkenyl and C7-C,Oheteroaralkenyl.
More preferred compounds of the formula laA are those in which X is -C(O)-, -C(S)-,
-C(O)O- or -C(S)O-; R3 is hydrogen or My where y and M are as defined above; R4 is
CH2-C6Hs, (CH2)2-C6Hs, cyclohexyl, methyl, ethyl or isopropyl which are unsubstituted or
substituted by one or more substituents selected from the group consisling of NH2,
C3-Cl2cycloalkyl, primary amino, secondary amino, sulfonamido, carbamido and amino-
carbonylamido; RT' is C,-C,2alkyl, which is unsubstituted or substituted by one or more
C(O)ORsl, where R5l is as defined above; and RT2 is C3-C,2cycloalkyl, C2-C"heterocyclo-
.
CA 022608~4 l999-0l-l2
W 098/06730 PCT~EPg7/04279 -12-
alkyl, C6-C10aryl or C5-Cgheteroaryl, which are unsubstituted or substituted by one or more
substituents selected from the group consisli,)g of OH, C,-C,2alkyl, C6-C,0aryl or
C5-Cgheteroaryl.
Most preferred compounds of the formula laA are those in which X is -C(O)- or-C(O)O-; R3
is hydrogen or My where y and M are as defined above; R4 is R'4; RT' is C1-C,2alkyl, which is
substituted by C(O)ORs', where Rs' is as defined above; and RT2 is -3,5-(OH)2C6H3,
-3,4-(OH)2C6H3, -3,4-(OCH3)2C6H3, -2-(OH)C6H4 or thyminyl.
Especially preferred compounds of the formula laA are those in which X is -C(O)- or
-C(O)O-; R3 is hydrogen or My; R4 is CH2-c6Hs~ (cH2)rc6H5~ cyclohexyl, methyl, ethyl or iso-
propyl which are unsubstituted or substituted by one or more substituents selected from the
group consisting of NH2, cyclohexyl, C6-C,Oaryl, R3C(o)N(R9)-, RBS(0)2N(R9)-,
R8NHC(O)NR9-, NR9C(o)NHR'3 and R3R9N-, in which R8, R9, R3 and R9 are, independently
of one another, hydrogen, C,-C12alkyl, cyclohexyl, phenyl, naphthyl or C7-C"aralkyl, which
are unsubstituted or substituted by one or more suhstituents selected from the group con-
sisting of OH, F, Cl, C(O)ONa, nitro, cyano, SO3Na, C1-C6alkyl, methoxy and phenyl; RT1 is
C,-C12aikyl, which is sllhstituted by C(O)OC1-C12alkyl; and RT2 is -3,4-(OH)2C6H3 and
3,4-(OCH3)2C6H3
Among these compounds of the formula laA those are preferred wherein X is -C(O)- or
-C(O)O-; R3 is hydrogen, K or Na; R4 is R"4; RT1 is CH3; and RT2 is -3,4-(OH)2C6H3 or
-3,4-(OCH3)2C6H3.
A second preferred embodiment of the present invention relates to compounds of the
formula IB
OH
R300C~o '~ o" ~~C~
HO ~
'- ~,oH--R~5 (IB)
R2 OH
HO
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
- 13-
in which R2, R3, R4 and RB5 have the above meanings.
In a particular embodiment of the compounds of the formula IB, RB~ corresponds to a group
of the formula llaB or llbB
o
NRe6Rs7 (llaB), --NyRB" (llbB)
o
in which
RB6 is hydrogen, C,-C,2alkyl, C3-C12alkenyll C3-C12cycloalkyl, C3-C12cycloalkenyl,
C2-C11heterocycloalkyl, C2-C"heterocycloalkenyll C6- or C,0aryll Cs-c9heteroaryl~ C7-C1tar-
alkyl, C6-C,0heteroaralkyl, Cg-C,,aralkenyl or C8~C10heteroaralkenyl;
F~B7 is C1-C,2alkyl, C3-C,2alkenyll C3-C,2cycloalkyll C3-C,2cycloalkenyll C2-C"heterocyclo-
alkyl, C2-C"heterocycloalkenyll C6- or C1Oaryll Cs-c9heteroaryll C7-C"aralkyll C6-C,Ohetero-
aralkyl, Cg-C"aralkenyl, C8-C,0heteroaralkenyll C(O)ORs'l C(O)R~8l SO2R1~ or SC3M
wherein R38 is hydrogen, C(O)OR5'1 C,-C12alkyll C3-C,2alkenyll C3-C12cycloalkyllC~-C12cycloalkenyl, C2-C1,heterocycloalkyl, C2-C"heterocycloalkenyl, C6- or C,Oaryl,
Cs-Cgheteroaryl~ C7-C"aralkyl, C6-C,Oheteroaralkyl, Cg-C"aralkenyl, C8-C,0heteroaralkenyl,
primary amino or secondary amino; Rs'l R'~, y and M are as defined above; and
R~11 is C2-C4alkylene, C2-C4alkenylene, 1l2-C3-C,2cycloalkylenel 1l2-C3-C,2cycloalkenylene,
1 ,2-C2-C, 1 heterocycloalkylene, 1 ,2-C2-C"heterocycloalkenylenel 1 l2-C6- or C,Oarylene,
1,2-Cs-Cgheteroarylene, 1l2-C8-C"aralkylene or 1l2-C6-C,0heteroaralkylene; and
alkyl, alkenyll cycloalkyll cycloalkenyll heterocycloalkyll heterocycloalkenyll aryll heteroaryll
aralkyl, heteroaralkyl, aralkenyl and heteroaralkenyl are unsubstituted or substituted by one
or more substituents selected from the abovementioned group of substituents.
Preferred compounds of the formula IB are those in which R3 is Hl K or Na.
Preferred compounds of the formula IB are those compounds in which R4 is R4bl R4b being
C1-C,2alkyl, C3-C,2cycloalkyl or C2-C"heterocycloalkyll where alkyll cycloalkyl and hetero-
cycloalkyl are unsubstituted or substituted by one or more substituents as defined above,
preferably R4 is optionally substituted C,-C6alkyll more preferably methyl substituted by
. .,
CA 022608~4 l999-0l-l2
W O 98/06730 PCT~EP97/04279
-14-
C3-C12cycloalkyl. Particularly preferred compounds of the formula IB are those compounds
in which R4 is cyclohexyl-methyl.
In a particular embodiment of the invention RB5 is primary amino or amido, preferably amido.
Preferably RB5 corresponds to a group of the formula llaB or llbB, in which RB6 is hydrogen,
C,-C12alkyl, C3-C,2cycloalkyl, C2-C11heterocycloalkyl, C6- or C10aryl, C5-Cgheteroaryl,
C7-C"aralkyl or C6-C,0heteroaralkyl; RB7 is C,-C,2alkyl, C3-C12cycloalkyl, C2-C1,heterocyclo-
alkyl, C6- or C10aryl, C5-C9heteroaryl, Crc11aralkyl~ C6-C,0heteroaralkyl, C(O)OR's', C(o)RB3,
SO2R1~ or SO3My~ wherein R'~, y and M are as defined above, R's' is My~ C1-C12alkyl,
C3-C12cycloalkyl, C2-C11heterocycloalkyl, C6- or C10aryl, Cs-c9heteroaryl~ Crc~1aralkyl or
C6-C10heteroaralkyl; R~3 is hydrogen, C(O)ORs', C1-C,2alkyl, C3-C,2cycloalkyl, C2-C1,hetero-
cycloalkyl, C6- or C,Oaryl, Cs-Cgheteroaryl, C7-C11aralkyl, C6-C,0heteroaralkyl, primary amino
or secondary amino; and RB" is C2-C4alkylene, 1,2-C3-C,2cycloalkylene or 1,2-C6- or
C,Oarylene; and alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl and heteroaralkyl
are unsubstituted or suhstih~ted by one or more substituents as defined above. Preferably
RBs corresponds to a group of the formula ItaB or llbB, in which Rr~6 is hydrogen, C1-C,2alkyl,
C3-C,2cycloalkyl or C6- or C10aryl; RB' is C,-C12alkyl, C3-C,2cycloalkyl, C6- or C10aryl,
C(O)OR"s', C(o)R'B3, SO2R"~ or SO3My~ wherein R"s' is My~ C,-C12alkyl, C3-C12cycloalkyl or
C6- or C,Oaryl; R'B~ is hydrogen, C(O)OR"s', C,-C,2alkyl, C3-C,2cycloalkyl or C6- or C1Oaryl,
primary amino or secondary amino; R"~ is C1-C12alkyl, C3-C12cycloalkyl or C6- or C10aryl; and
RB" is 1,2-C6- or C10arylene; and alkyl, cycloalkyl and aryl are unsubstituted or sl~hstihlted
by one or more substituents selected from the group consisting of halogen, nitro, C,-
C,2alkyl, C,-C,2alkoxy, C3-C,2cycloalkyl or C6- or C,Oaryl; and My is K or Na. More preferably
RBs corresponds to a group of the formula llaB, in which RB6 is hydrogen, C,-C,2alkyl, C3-
C,2cycloalkyl or C6- or C10aryl; RB7 is C,-C12alkyl, C3-C12cycloalkyl, C6- or C1Oaryl, C(O)OR''s',
C(o)R'r~3, SO2R"~ or SO3My~ wherein R"s', R'B3, R"~, y and M are as defined above; and
alkyl, cycloalkyl and aryl are unsubstituted or substituted by one or more substituents
selected from the group consisting of halogen, nitro, C,-C12alkyl, C,-C,2alkoxy,C3-C12cycloalkyl or C6- or C10aryl. Most preferably RB5 corresponds to a group of the formula
llaB, in which RB6 is hydrogen or C1-C12alkyl; RB7 is C,-C12alkyl, C(O)OC1-C,2alkyl, C(O)RB~,
SO2C6- or C,Oaryl or SO3My; RBi3 is C(O)OMy~ C1-C12alkyl, C3-C,2cycloalkyl, C6- or C10aryl or
primary amino; and alkyl, cycloalkyl and aryl are unsubstituted or substituted by one or more
substituents selected from the group consisting of halogen, nitro, C,-C12alkyl, C1-C,2alkoxy
CA 022608~4 l999-0l-l2
W O 98/06730 PCT~EP97/04279
-15-
or C6- or C,Oaryl. Especially preferred compounds are those in which RBs corresponds to a
group of the formula llaB, in which R~5 is hydro9en, methyl or benzyl; R~7 is methyl, benzyl,
~ C(O)ORs1a, C(O)RB8a, SO2R'~a or SO3Na, wherein Rsl~ is methyl or methyl substituted with
one or more substituents selected from phenyl, phenyl sl~hstitl~ted with one or more substi-
tuents selected from methoxy and nitro, and naphthyl; RB8a is C(O)ONa, methyl s' lhstituted
with one or more phenyl, ethyl substituted with phenyl, cyclohexyl, phenyl, phenyl
substituted with one or more substituents selected from methoxy, chlorine, nitro, phenyl and
trifluormethyl, naphthyl, NH(CH2)2COONa, NHc6Hs or NHCH2CH3; and R10a is tolyl.
Most preferably R~s is R'B5, R'~5 being -NHC(O)CH2C6H5, -NHC(O)CH(C6H5)2, -NHSO3Na,
-NHC(O)(CH2)2C6H5, -NHc(o)c6H11l -NHc(o)c6H5l -NHc(o)c6H4(4-ocH3)l -NHCH2C6H5,
-NHC(O)C6H3(3,4-OCH3)2, -NHC(O)C6H4(4-CI), -NHC(O)C6H4(4-NO2), -NHC(O)C6H4(4-C6H5),
-NHC(O)C6H4(4-CF3), -NHC(O)COONa, -NHC(0)-2-naphthyl, -NHC(0)-1 -naphthyl,
-NHC(O)NH(CH2)2COONa, -NHC(O)NHC6H5, -NHC(O)NHCH2CH3; -NHC(O)OCH2C6H5,
-NHC(O)OCH2C6H2(4,~-OCH3)2(2-NO2), -NHc(o)ocH2c6H4(4-No2)l -NHso2c6H4(4-cH3)l
-NHC(O)OCH2-2-naphthyl, -NHCH3, -N(CH2C6Hs)2, -N(cH3)c(O)c6Hs~ -N(CH2C6H5)C(O)C6H5
or -phthalimido.
Especially preferred as R~5 are -NHC(O)CH(C6H5k, -NHC(O)C6H,1, -NHC(O)C6H4(4-C6Hs),
-NHC(O)C6H5, -NHC(O)C6H4(4-OCH3), -NHc(o)c6H3(3l4-ocH3)2l -NHc(o)c6H4(4-cl)
-NHC(O)C6H4(4-NO2), -NHC(0)-2-naphthyl, -NHC(O)NHC6H5, -NHC(O)OCH2C6H5,
-NHSO3Na, -NHCH2C6Hs or -N(CH2C6Hs)2
A particularly preferred embodiment of the invention comprises compounds of the formula
laB
OH
~HO ~~ oX1~
- (laB)
C ~-J OH
HO
in which R3 is hydrogen or My and Rr~5 is a group of formula llaB or llbB as defined above.
. .
CA 022608~4 l999-0l-l2
W 098/06730 PCT~EP97/04279 -16-
Preferred compounds of the formula laB are those in which R3 is H, K or Na; RB6 is hydro-
gen, C,-Cl2alkyl, C3-C12cycloalkyl, C2-C,1heterocycloalkyl, C6- or C10aryt, Cs-Cgheteroaryll
C7-C"aralkyl or C6-C,Oheteroaralkyl; RB' is C1-C12alkyl, C3-C12cycloalkyl, C2-C11heterocyclo-
alkyl, C6- or C,Oaryl, Cs-Cgheteroaryl, C,-C11aralkyl, C6-C10heteroaralkyl, C(O)ORsl, C(O)RBa,
SO2R'~ or SO3Myl wherein R's', R'~, y and M are as defined above; Rr~8 is hydrogen,
C(O)OR's', C,-C,2alkyl, C3-C,2cycloalkyl, C2-C"heterocycloalkyl, C6- or C,Oaryl, C5-Cghetero-
aryl, C,-C"aralkyl, C6-C,Oheteroaralkyl, primary amino or secondary amino; and RB" is
C2-C4alkylene, 1 ,2-C3-Cl2cycloalkylene or 1,2-C6- or C,Oarylene; and alkyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, aralkyl and heteroaralkyl are unsubstituted or substituted
by one or more substituents as defined above.
More preferred compounds of the formula laB are those in which R3 is H, K or Na; Rr~6 is
hydrogen, C,-C12alkyl, C3-C,2cycloalkyl or C6- or C,Oaryl; RB7 is C,-C,2alkyl, C3-C,2cycloalkyl,
C6- or C,Oaryl, C(O)OR''s', C(O)R'B8, S02R"~ or SO3Myl wherein R"s', R'B6, R"~, y and M are
as defined above; and RB" is 1,2-C6- or C,Oarylene; and alkyl, cycloalkyl and aryl are
unsubstituted or substituted by one or more substituents selected from the group consisting
of halogen, nitro, C,-Cl2alkyl, C,-C,2alkoxy, C3-C,2cycloalkyl or C6- or C,Oaryl.
Most preferred compounds of the formula laB are those in which R3 is H, K or Na; RB6 is
hydrogen, C,-C,2alkyl, C3-C,2cycloalkyl or C6- or C,Oaryl; RB7 is C,-C,2alkyl, C3-C,2cycloalkyl,
C6- or C,Oaryl, C(O)OR''s', C(O)R'B8, SO2R"~ or SO3Myl wherein R"s', R'B6, R"~, y and M are
as defined above; and alkyl, cycloalkyl and aryl are unsuhstituted or substituted by one or
more substituents selected from the group consisting of halogen, nitro, C,-C,2alkyl,
Cl-C,2alkoxy, C3-C,2cycloalkyl or C6- or C,Oaryl.
Especially preferred compounds of the formula laB are those in which R3 is H, K or Na; RB6
is hydrogen or C,-C,2alkyl; RB' is C,-C,2alkyl, C(O)OC,-C,2alkyl, C(o)RB3, SO2C6- or C,Oaryl
or S03My, wherein RB8 is C(O)OMy~ C,-C,2alkyl, C3-C,2cycloalkyl, C6- or C10aryl or primary
amino; and alkyl, cycloalkyl and aryl are unsubstituted or substituted by one or more
substituents selected from the group consisting of halogen, nitro, C,-C12alkyl, C,-C,2alkoxy
or C6- or C,Oaryl.
CA 022608~4 1999-01-12
W 098/06730 PCT/EP97/04279 -17-
Especially preferred compounds of the formula laB are those in which R3 is H, K or Na; Rr~6
is hydrogen, methyl or benzyl; RB7 is methyl, benzyl, C(O)ORsla, C(O)R~, SO2Rl~a or
~ SO3Na, wherein Rsla, R~3a and R'~a are as defined above.
A third preferred embodiment of the present invention relates to compounds of the formula
IC
OH
R300C~,o, ~ o" ~
RHO~'~O ~O
o~ (IC)
R OH
HO
in which R2, R3, R4 and Rs have the above meanings.
Preferred compounds of the formula IC are those in which R3 is H, K or Na.
Preferred compounds of the formula IC are those compounds in which R4 is R4a as defined
above.
In a particular embodiment of the invention R5 is X'-RT'C, C(o)NRT2CRT3c or C(o)oRT5C,
wherein X' is Cl-C4alkylene and RT'C, RT2C, RT3C and RTsC are as defined above.
Preferably R5 is X'-RT1C or C(O)ORTsC, wherein X', RT'C and RTsC are as defined above.
More preferred are compounds of the formula IC wherein Rs is X'-RT'Ca or C(o)oRT5C,
wherein X' and RT5C are as defined above and RT'Ca is hydrogen or ORT6C wherein RT6C is
as defined above.
Most preferred are compounds of the formula IC wherein Rs is X'-RT'C or C(o)oRT5C,
wherein X' is C1-C4alkylene, RT1C is hydrogen or OH; and RTsC is hydrogen or My~ Preferably
Rs is CH20H, CH3 or C(O)ONa.
CA 022608~4 l999-0l-l2
W O 98/06730 PCTAEP97/04279
-18-
Particularly preferred compounds of the formula IC are compounds of the formula laC
OH
R300C~O, ~ O" ~
O~ HO ~ - ~ ~~ (laC)
o~,OH
H3C ~ OH
HO
in which R is hydrogen, K or Na; and R5 is X'-RT'C C(o)NRT2CRT3c or C(o)oRT5C h
X~ RT1c RT2C RT3c and RT5C are as defined above-
More preferred are compounds of the formula laC wherein R3 is hydrogen, K or Na; and R5
X~ RT1Ca c(O)ORT5ca wherein X~ RT~Ca and RT5C are as defined above-
Most preferred are compounds of the formula laC wherein R3 is hydrogen, K or Na; and Rs
is X'-RT'C or C(o)oRT5C, wherein X' is C1-C4alkylene, RT'C is hydrogen or OH; and RT5C is
hydrogen or My More preferably R5 is CH20H, CH3 or C(O)ONa.
The present invention also comprises a process for the preparation of the compounds of
the formula I wherein the corresponding galactose-GlcNAc-disaccharide or gAIActose-tetra-
hydropyran dimer is linked with the corresponding fucose-derivative or the corresponding
fucose-GlcNAc-disaccharide or fucose-tetrahydropyran dimer is iinked with the correspond-
ing galactose, wherein the groups R', RT', X-RT2, R~5 and/or Rs are optionally introduced
before or after the formation of the dimer or trimer. Where required, one or more protecting
groups are removed and the compounds thus obtained are converted into salts.
In a preferred process for the preparation of the compounds of the formula IA the corres-
ponding galactose-GlcNAc-disaccharide is linked with the corresponding fucose-derivative
or the corresponding fucose-GlcNAc-disaccharide is iinked with the corresponcling galac-
tose wherein the groups R', RT' and X-RT2 are optionally introduced before or after the
formation of the dimer or trimer.
CA 022608~4 l999-0l-l2
W O 98/06730 PCT~EP97/04279
-19-
Typically the process for the preparation of the compounds of the formula IA comprises
(A1 ) reacting a compound of the formula IIIA
oJ~,~OR
- R~20~ oR60 (IIIA)
oR-2
wherein each Rl2 independently is hydrogen or a protecting group, R60 is R' or a protecting
group and R's is a leaving group, with a compound of the formula IVA
oR6-
o~NHR
R~20~ oR63 (IVA)
R640
wherein Rt2 is as defined above, R6' is RT' or a protecting group, or OR61 is R'5, R62 ishydrogen, a protecting group or X-RT2, R63 is hydrogen or a protecting group and R64 is
hydrogen or a protecting group or R12 and R64 together form a protecting group, and
(A2) reacting the resulting disaccharide with a compound of the formula VA
R1s
,0~ oR'2 (VA)
2 ~ 12
oR12
wherein R2, R12 and R'~ are as defined above; wherein the groups R', RT' and X-RT2 are
optionally introduced before or after step (A1) or step (A2); and, where required, removing
the protecting groups; or
(B1) reacting a compound of the formula VA with a compound of the formula IVA, and
(B2) reacting the resulting disaccharide with a compound of the formula IIIA; wherein the
groups R', RT' and X-RT2 are optionally introduced before or after step (B1 ) or step (B2);
and, where required, removing the protecting groups.
For example, a compound of formula la may be prepared by reacting a compound of
formula IVA with RT'-Otl, followed by a reaction with a compound of formula VA. The
resulting compound is reacted with RT2-X-R'4, wherein R'4 is a leaving group, then with a
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
-20-
compound of formula IIIA and finally with R'-R'3, wherein R13 is a leaving group. Where
required, the protecting groups are removed and the compounds of formula IA are
converted into salts.
As it will be appreciated, this reaction scheme is an example and may be carried out in a
different sequence to produce a compound of formula IA.
The compounds of the formula IIIA, IVA and VA are known or may be obtained in
accordance with methods known and practiced in the art.
Hydroxy protecting groups are generally known in the sugar and nucleotide chemistry and
are described, for example, by Greene and Wuts [Protective Groups in Organic Synthesis,
Wiley, New York (1991)]. Examples of such protecting groups are: linear and branched
C,-C8alkyl, in particular C,-C4alkyl, for example methyl, ethyl, n- and i-propyl, n-, i- and
t-butyl; benzyl, methylbenzyl, dimethylbenzyl, methoxybenzyl, dimethoxybenzyl, bromo-
benzyl, 2,4-dichlorobenzyl; diphenylmethyl, di(methylphenyl)methyl, di(dimethylphenyl)-
methyl, di~methoxyphenyl)methyl, di(dimethoxyphenyl)methyl, triphenylmethyl, tris-4,4',4"-
tert-butylphenylmethyl, di-p-anisylphenylmethyl, tri~methylphenyl)methyl, tri(dimethylphenyl)-
methyl, methoxyphenyl(diphenyl)methyl, di(methoxyphenyl)phenylmethyl, tri(methoxy-
phenyl)methyl, tri(dimethoxyphenyl)methyl; triphenylsilyl, alkyldiphenylsilyl, dialkylphenylsilyl
and trialkylsilyl with 1 to 20, preferably 1 to 12, and particularly preferably 1 to 8 C atoms in
the alkyl groups, for example triethylsilyl, tri-n-propylsilyl, i-propyl-dimethylsilyl, t-butyl-di-
methylsilyl, t-butyl-diphenylsilyl, n-octyl-dimethylsilyl, (1,1,2,2-tetramethylethyl)dimethylsilyl;
C2-C,2-, in particular C2-C~acyl, such as acetyl, propanoyl, butanoyl, pentanoyl, hexanoyl,
benzoyl, methylbenzoyl, methoxybenzoyl, chlorobenzoyl and bromobenzoyl; substituted
methylidene groups which are obtainable by acetal or ketal formation from adjacent
hydroxyl groups of the sugars or sugar derivatives with aldehydes and ketones, which pre-
ferably contain 2 to 12 or 3 to 12 C atoms, for example C,-C,2alkylidene, preferably
C,-C6alkylidene and in particular C,-C4alkylidene, such as ethylidene, 1,1- and 2,2-propyl-
idene, 1,1- and 2,2-butylidene, benzylidene. The protecting groups may be identical or diffe-
rent.
Preferably R12 and R64 together form an alkylidene group with, preferably 1 to 12 and, more
preferably 1 to 8 C atoms. These protecting groups may be removed under neutral or weak-
CA 022608~4 1999-01-12
W 098/06730 PCT/EP97104279
-21-
ly acidic conditions. R12 and R64 are, particularly, together alkylidene, for example unsubsti-
tuted or alkyl- or alkoxy- substituted benzylidene.
Compounds of the formula IB wherein RB5 is a group of formula llaB may be produced by
reacting a compound of the formula IVB
OH
-OOC~O"~ O", ~
R HO''~O 0~~
' NH + (IVB)
' ~ OH
HO
wherein R2 and R4 have the abovementioned meanings
(a) in the case where RB6 is hydrogen and
(al) F~B7 is C~-C12alkyl, C3-C12alkenyl, C7-C~aralkyl, C6-ClOheteroaralkyl, Cg-C~laralkenyl~
C8-C10heteroaralkenyl, which are unsubstituted or substituted by one or more substituents,
with an aldehyde of formula VB
RB7 CHO (VB)
wherein RB7 is hydrogen, C1-C1~alkyl, C2-C11alkenyl, C3-C~2cycloalkyl, C3-C12cycloalkenyl,
C2-C1,heterocycloalkyl, C2-C,lheterocycloalkenyl, C7-C,0aralkyl, C6- or C10aryl,C6-Cgheteroaralkyl, C5-Cgheteroaryl, C8-C~0aralkenyl or C7-Cgheteroaralkenyl, which are
unsubstituted or substituted by one or more substituents; or
(a2) RB7 is C1-C12alkyl, C3-C,2alkenyl, C3-Cl2cycloalkyl, C3-Cl2cycloalkenyl, C2-Cl~hetero-
cycloalkyl, C2-C~1heterocycloalkenyl, C7-C11aralkyl, C6-C10heteroaralkyl, Cg-C11aralkenyl,
C8-C10heteroaralkenyl, which are unsubstituted or substituted by one or more substituents,
with a ketone of formula VlaB or VlbB
RB7CoR8r (VlaB) O=CRB12 (VlbB)
wherein each of RB' and RBT independently is C,-C"alkyl, C2-C"alkenyl, C3-Cl2cycloalkyl,
C3-C,2cycloalkenyl, C2-C,1heterocycloalkyl, C2-C,1heterocycloalkenyl. C7-C,0aralkyl, C6- or
Cl0aryl, C6-Cgheteroaralkyl, Cs-Cgheteroaryl~ C~-C,Oaralkenyl. Crc9heteroaralkenyl~ which
are unsubstituted or substituted by one or more substituents; and RB'2 is C3-C,0alkylene or
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
- 22 -
C3-C,Oalkenylene, for example cyclobutanon, cyclodecanon, cyclobutenon and cyclodecen-
on, which are unsubstituted or substituted by one or more substituents; or
(a3) RB' is C(O)ORs', C(o)Rr~5 or SO2R'~, wherein Rs' and R'~ are as defined above and RB5
is hydrogen, C(O)ORs', C1-C12alkyl, C3-C,2alkenyl, C3-Cl2cycloalkyl, C3-C,2cycloalkenyl,
C2-C"heterocycloalkyl, C2-C11heterocycloalkenyl, C6- or C10aryl, Cs-c9heteroaryl~ C7-C11ar-
alkyl, C6-C10heteroaralkyl, Cg-Cl1aralkenyl or C8-C10heteroaralkenyl; and alkyl, alkenyl, cyclo-
alkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, aralkyl, heteroar-
alkyl, aralkenyl and heteroaralkenyl are unsubstituted or substituted by one or more substi-
tuents,
with a compound of formula VIIB
R13R'3T (VIIB)
is C(O)OR , C(O)R~8 or S02R1~, wherein R51 RB8 d R10
above; and R13 is a leaving group; or
(a4) RB7 is C(o)RB5, wherein RB5 is primary amino or secondary amino; with an isocyanate
OCNR8 (VIIIB)
wherein R~ is hydrogen, S02R1~, OSO2R1~, C1-C12alkyl, C3-C12cycloalkyl, C2-C11heterocyclo-
alkyl, C6- or C,Oaryl, C5-Cgheteroaryl, C7-C1,aralkyl, C6-C,Oheteroaralkyl, C8-C16aralkenyl,
which are unsubstituted or substituted by one or more substituents;
(a5) RB7 is SO3Myl wherein My has the abovementioned meanings, with a complex offormula IXB
SO3~ NC5H5 (IXB);
(b) in the case where R~6 is C,-C,2alkyl, C3-C12alkenyl, C7-C"aralkyl, C6-C10heteroaralkyl,
Cg-C,1aralkenyl or C5-C10heteroaralkenyl; arld
(bl) RB7 is C,-C,2alkyl, C3-C12alkenyl, C3-C,2cycloalkyl, C3-C12cycloalkenyl, C2-C,1hetero-
cycloalkyl, C2-C"heterocycloalkenyl,C,-C"aralkyl, C6-C10heteroaralkyl, Cg-C1,aralkenyl or
C8-C,Oheteroaralkenyl, which are unsubstituted or substituted by one or more substituents
subsequently with an aldehyde of formula VB or a ketone of formula VlaB or VlbB;(b2) R~7 is C(O)ORs1, C(o)RB5 or SO2R1~, wherein R51 is hydrogen, My~ C1-C,2alkyl,
C3-C,2alkenyl, C3-C12cycloalkyl, C2-C11heterocycloalkyl, C6- or C10aryl, C5-Cgheteroaryl,
C,-C1,aralkyl or C6-C,Oheteroaralkyl; RB5 is hydrogen, C(O)ORs1, C1-C12alkyl, C3-C12alkenyl,
C3-C,2cycloalkyl, C3-C,2cycloalkenyl, C2-C"heterocycloalkyl, C2-C11heterocycloalkenyl, C6-
or C10aryl, Cs-Cgheteroaryl, C7-C,1aralkyl, C6-C10heteroaralkyl, Cg-C11aralkenyl or
C8-C,Oheteroaralkenyl; R10 is C,-C,2alkyl, C3-C12cycloalkyl, C2-C11heterocycloalkyl, C6- or
C,Oaryl, Cs-Cgheteroaryl, C7-C1,aralkyl or C6-C10heteroaralkyl, which are unsubstituted or
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
-23-
substituted by one or more substituents; subsequently with an aldehyde of formula VB and
a compound of formula VIIB; or
(b3) RB7 is C(O~RB8, wherein R~8 is primary amino or secondary amino; subsequently with an
aldehyde of formula VB and a compound of formula VIIIB;
(b4) RB' is SO3My~ subsequently with an aldehyde of formula VB and a compound offormula IXB.
Compounds of formula IB wherein RB5 is a group of formula llaB wherein RB7 is C6- or
C10aryl or Cs-Cgheteroaryl, may be produced by reductively aminating a compound of
formula IVbB
oR12
R300C~,o, ~o "~
R120
R120i CHO (IVbB)
~--1' oR'2
12
R120
wherein R2, R3 and R4 are as defined above and R12 is hydrogen or a protecting group with
an aromatic amine, optionally removing the protecting groups, and further reacting the
resulting compound as described in (b) above.
Compounds of formula IB wherein RB5 is a group of formula llbB may be produced by
reacting a compound of the formula IVB wherein R2, R3 and R4 are as defined above with a
compound of formula VllbB
o
R'~RB1 (VllbB)
O
wherein RB11 has the abovementioned meanings; and each R'3 is independently a leaving
group.
CA 022608~4 1999-01-12
WO 98106730 PCTIEP97/04279
- 24 -
Leaving groups may be: halides, such as chloride, bromide and iodide, and oates for
example of the formula RB~-O (in which case formula VliB is an anhydride RBT-O-RBT) or
alkoxides (alkylO ).
The compounds of the formula VB to IXB are known or may be obtained by known
methods.
The compounds of the formula IVB and IVbB are novel and form part of the present inven-
tion. They may be obtained starting from commercially available 3,4,6-triacetoxyglucal by
(a) deprotecting said compound, protecting its 6-position, coupling with a suitably protected
and activated galactose, replacing the 6-protecting group by a leaving group, substituting
with an N-nucleophile, coupling with a suitably protected and activated L-fucose, introducing
the group -CH(CooRB8)R4, in which R~8 is a carboxylate protecting group and R4 has the
abovementioned meanings, into the prior deprotected g~ctose residue, reducing the
glucal residue, removing the fucose protecting groups and optionally reducing the residue
R4; or
(b) reducing said compound, deprotecting and converting the 6-position into a leaving
group, coupling with a suitably protected and activated g~lactose, coupling with a suitably
protected and activated L-fucose, substituting the 6-position with an N-nucleophile, intro-
ducing the group -CH(CooRBa)R4 into the prior deprotected g~l~ctose residue and remov-
ing the protecting groups; or
(c) reducing said compound, deprotecting and protecting its 4- and 6-position, coupling with
a suitably protected and activated g~l~ctose, removing the 4- and 6-protecting groups, con-
verting the 6-position into a leaving group, substituting with an N-nucleophile, coupling with
a suitably protected and activated L-fucose, introducing the group -CH(CooR88)R4 into the
prior deprotected galactose residue, reducing the glucal residue and removing the fucose
protecting groups; or
(d) deprotecting said compound, converting the 6-position into a leaving group, substituting
with an N-nucleophile, coupling with a suitably protected and activated g-'-ctose, coupling
with a suitably protected and activated L-fucose, introducing the group -CH(CooRe8)R4 into
the prior deprotected galactose residue, reducing the glucal residue, removing the fucose
protecting groups and optionally reducing the residue R4; or
(e) deprotecting said compound, protecting its 6-position, coupling with a suitably protected
and activated galactose, reducing the glucal double bond, converting the 6-position into a
CA 022608~4 1999-01-12
W 098/06730 PCT~EP97/04279 -25-
leaving group, substituting with an N-nucleophile, coupling with a suitably protected and
activated L-fucose, introducing the group -CH(CooR~)R4 into the prior deprotected galac-
tose residue, reducing the glucal residue and removing the fucose protecting groups; or
(f) deprotecting said compound, protecting its 6-position, coupling with a suitably protected
and activated galactose and subsequently with a suitably protected and activated L-fucose,
introducing the group -CHtCooRBg)R4 into the prior deprotected galactose residue, pro-
tecting the remaining g~lactose hydroxy groups, deprotecting the 6-protecting group of the
glucal and oxidizing said position.
The abovementioned strategies (a) to (e) may for example be performed by using a suitably
protected and activated galactose which already contains the group -CH(CooRB8)R4 This
compound may for example be obtained starting from an activated galactose by introducing
a protecting group at the anomeric position, deprotecting said compound, introducing the
group -CH(CooR~&)R4 protecting the residual hydroxyl groups, deprotecting and activating
the anomeric position.
Suitable activating groups for sugars and glycosylation are known to the person skilled in
the art and are described for example by Toshima and Tatsuta [Chem. Rev. 93:1503(1993)]; Paulsen [Angew. Chem. Int. Ed. Engl. 21:1~ (1982)] and Schmidt and Kinzy [Adv.
Carbohydr. Chem. Biochem. 50:21 (1994)].
Examples for N-nucleophiles are NaN3, NH3, primary amines and secondary amines, pre-
ferably the N-nucleophile is NaN3.
Suitable reducing conditions are for example H2, Pd/C 10%, MeOH; H2, Pd(OH)2/C 10%,
dioxane/water 2/1; or H2, Rh/AI2O3 5%, dioxane/water 2J1.
The compounds of formula IVB, IVbB and VB, VIB, VIIB, VIIIB and IXB respectively may be
employed in equimolar amounts or, advantageously, in excess, for example in an amount
which is up to ~ times, preferably 2 times the amount of the compound of formula IVB or
IVbB.
CA 022608~4 1999-01-12
W 098/06730 PCT~EP97/04279 -26-
Examples of carboxylate protective groups are esters preferably methyl and benzyl esters.
Methyl esters are preferably cleaved under the abovemenlionetl basic conditions and
benzyl esters are preferably cleaved under the abovementioned reducing conditions.
Compounds of formula IC may be prepared by converting a compound of the formula IIC
oR-2
R300c~o, ~ o"
R4 ~O ~O
oR.2 OH (IIC)
'~ 12
R-20
wherein R2 and R4 have the abovementioned meanings R3 has the meanings of R3 or is a
protecting group and R12 means a protecting group applying procedures known in the art.
Examples ot such conversions are e.g.
(A) in the case that Rs is X-RT'C wherein X is methylene and RT'C is
(a) hydrogen: removing the alcohol functionality;
(b) halogen: transforming the alcohol function into a halide function;
(c) C,-C,2alkyl, C,-C1,heteroalkyl, C3-C,2alkenyl, C3-C,2cycloalkyl, C3-C,2cycloalkenyl,
C2-C"heterocycloalkyl C2-C"heterocycloalkenyl C6- C,0- or C,4aryl C2-C9heteroaryl
C7-C,laralkyl C6-C,0heteroaralkyl Cg-C,,aralkenyl or CB-C,0heteroaralkenyl: oxidizing the
alcohol reacting the resulting aldehyde with a suitable C-nucleophile and removing the
resulting secondary alcohol functionality;
(d) ORT6C wherejn RT6c is
(a) hydrogen: removing the protecting groups;
(~) C,-C,2alkyl C,-C"heteroalkyl C3-C,2alkenyl C3-C,2cycloalkyl C3-C,2cycloalkenyl
C2-C"heterocycloalkyl C2-C"heterocycloalkenyl C6- C,0-orC,4aryl C2-Cgheteroaryl
C,-C1,aralkyl C6-C,Oheteroaralkyl Cg-C,,aralkenyl or C8-C10heteroaralkenyl: forming the
corresponding ether;
(y) So3RT5C or Po3RT7CRT8c: introducing the S04- or PO4-function using a suitable S03- or
PO3-donor (eg. S03 ~ pyridine);
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
- 27 -
(~) C(O)ORT9C: reacting with a compound of formula Hal-C(O)-ORT9C;
(~) C(S)NRT2CRT3c or C(o)NRT2CRT3c: reacting with for exa",ple phosgene [C(O)CI2] or
thiophosgene [C(S)CI2], and substituting the resulting carbonic/thiocarbonic acid chloride
With HNRT2CRT3C;
(e) oC(o)RT4C: forming the corresponding ester;
(f) SRT4C, SO2RT9C or SO3RTsC: transforming the alcohol group into a leaving group, reacting
with a suitable S-nucleophile and optionally oxidizing the resulting SR group;
(B) in the case that R5 is X'-RT'C, wherein X' is Cz-C4alkylene: proceeding as in (A(c))
followed by one of the procedural variants A(a) to A(f);
(C) in the case that R5 is C(o)NRT2CRT3c: oxidizing the alcohol and reacting the resulting
carboxylic acid to form the corresponding amide;
(D) in the case that R5 is C(o)RT4C: oxidizing the alcohol and optionally reacting the resulting
aldehyde with a suitable C-nucleophile and oxidizing the resulting secondary alcohol func-
tionality to the corresponding ketone;
(E) in the case that R5 is C(o)oRT5C: oxidizing the alcohol and optionally reacting the result-
ing carboxylic acid to form the corresponding ester;
wherein each of the above variants (except of [A(d)(a)]) is followed by the removal of the
protecting groups.
The compounds of the formula IIC are new and form part of the present invention. They
may be produced by linking the corresponding g~lactose-1,2-dideoxyglucose-disaccharide
with the corresponding fucose-derivative or the corresponding fucose-1 ,2-dideoxyglucose-
disaccharide with the corresponding galactose wherein the group -CH(CooR3)R4 is
optionally introduced before or after the formation of the dimer or trimer. The compounds of
formula IIC may be obtained by following a procedure as r~isclosed for the compounds of
formula IA above, the group -CH(CooR3)R4 being introduced by reaction with
R'3-CH(CooR3 )R .
Leaving groups as R'3 may be a halide or unsubstituted or halogenated R-SO2-, in which R
is C,-C12alkyl, in particular C1-C6alkyl and mono-, di- or trifluoromethyl, Cs-c6cycloalkyl~
phenyl, benzyl, C1-C12alkylphenyl, in particular C,-C4alkylphenyl, nitrophenyl, or C,-C12alkyl-
benzyl, in particular C,-C4alkylbenzyl, for example methane, ethane, propane, butane,
CA 022608~4 1999-01-12
WO 98t06730 PCT/EP97/04279
- 28 -
benzene, benzyl- and p-methylbenzenesulfonyl. Preferred leaving groups are Cl, Br, I,
-SO2CF3 (triflate) and p-nitrobenzenesulfonyl, -SO2CF3 being more prefer"ad.
Leaving groups in the meaning of R'4 are for example halides, such as preferably chloride
and bromide, and especially in the case when X is -C(O)- carboxylates and groups of for
example the formulae
--O~F, --~~~2, N~ ~ and ~ wherein
R'~~ is for example isopropyl or cyclohexyl.
Examples of leaving groups which are especially useful in glycosylation reactions, i.e. here
in the meaning of Rl5 are -S-Ctl3, -S-CH2-CH3, -S-Ph, -O-C(=Ntl)-CCI3, -O-C(-O)-CH3,
OP(OR)2 and halogen for example Cl, Br and 1. These leaving groups can be in the axial or
in the equatorial position.
It has proved advantageous to activate the 3-OH group of the g~'~ctose residue prior to
etherification by stannylation. Particularly suitable for this purpose are dialkyltin oxides,
dialkyltin alkoxylates and bis(trialkyl)tin oxides. Some examples are dibutyltin oxide, dibutyl-
tin(O-methyl)2 and (tributyltin)20. The activating agents are preferably used in stoichiometric
amounts. In this case, the reaction is carried out in two stages, namely a) activation and b)
coupling with e.g. RT1-OH.
Further details of the preparation of the compounds of the formula IA, IB and IC are
described e.g. in the examples.
The compounds of formula I exhibit valuable pharmacological properties as indicated in
tests and are therefore indicated for therapy. In particular the compounds of formula I inhibit
the binding of E-selectin to SLea as disclosed in Example C1 and the interaction of E-
selectin and its natural ligand as disclosed in Example C2.
CA 022608~4 1999-01-12
WO 98/06730 PCT/EP97/04279
- 29 -
The compounds are accordingly indicated for preventing or treating conditions or diseases
which are mediated by the binding of selectin in cellular adhesion, e.g. acute or chronic
inflammatory or autoimmune dise~ses such as rheumatoid arthritis, asthma, allergy condi-
tions, psoriasis, contact dermatitis, adult respi,d~ory dioll~ss syndrome, inflammatory bowel
dise~se and ophthalmic inflammatory clise~-~es, infection diseases such as septic shock,
traumatic shock, thrombosis and inappropriate platelet aggregation conditions, cardiovas-
cular ~ise~ses such as heart attacks, reperfusion injury, multiple sclerosis and neoplastic
diseases including metAst~sis conditions, strokes and acute or chronic rejection of organ or
tissue transplants.
Acute and chronic rejection play a role in the transplantation of organs or tissues from a
donor to a recipient of the same species (allograft) or different species (xenograft). Among
such transplanted organs or tissues and given illustratively are heart, lung, combined heart-
lung, trachea, liver, kidney, spleen, pancreatic (complete or partial, e.g. Langerhans islets),
skin, bowel, or cornea or a combination of any of the foregoing.
For the above uses the required dosage will of course vary depending on the mode of admi-
nistration, the particular condition to be treated and the effect desired, In general, however,
satisfactory results are achieved at dosage rates of from 0.1 to about 100 mg/kg/day, admi-
nistered in 1, 2, 3, or 4 doses/day, or in sustained release form. Suitable daily dosages for
oral administration to larger mammals, e.g., humans, are generally about 50 to 1500 mg,
preferably in the order of from 200 to 800 mg. Unit dosage forms suitably comprise from
about 25 mg to 0.750 9 of a compound of the invention, together with a pharmaceutical
acceptable diluent or carrier therefor.
The compounds of formula I may be administered by any conventional route of admini-
stration, e.g. enterally, preferably orally, e.g. in the form of tablets or capsules, or par-
enterally e.g. in form of injectable solutions or suspensions.
Pharmaceutically acceptable salts are to be understood as meaning, in particular, the alkali
metal and alkaline earth metal salts, for example sodium, potassium, magnesium and
calcium salts. Sodium and potassium ions and their salts are preferred.
CA 02260854 1999-01-12
W O 98/06730 PCT/EP97/04279
-30-
ln accordance with the foregoing the present invention further provides:
(a) a compound of formula I or a pharmaceutically accel,table salt thereof for use as a
pharmaceutical;
(b) a method for preventing or treating disorders as indicated above in a subject in need of
such treatment, which method coi"prises ad",i"i~lering to said subject an effective amount
of a compound of formula I or a pharm~ceutic~lly acceptable salt thereof;
(c) a pharmaceutic~l composition comprising a pharmaceuticaliy effective amount of the
compound of formula I or a pharmaceutically acceptable salt thereof together with a
pharmaceutically acceptable diluent or carrier;
(d) a compound of formula I or a pharmaceutically acceptable salt thereof for use in the
manufacturing of a medicament for use in the method as in (b) above.
The compound may be administered alone or in combination with one or more other anti-in-
flammatory or immunosu~pressive agents, for example in combination with cyclosporin A
and analogs thereof, FK-506 and analogs thereof, rapamycin and analogs thereof, myco-
phenolic acid, mycophenolate mofetil, mizoribine, 15-deoxyspergualine, leflunomide,
steroids, cyclophosphamide, azathioprene (AZA), or anti-lymphocyte antibodies or immuno-
toxins such as monoclonal antibodies to leukocyte receptors, e.g. MHC, CD2, CD3, CD4, or
CD25; especially in combination with a T-cell suppressant, e.g., cyclosporin A or FK-506.
Such combination therapy is further comprised within the scope of the invention, e.g., a
method according to 1 above further comprising administration concomitantly or in
se~uence of a therapeutically or synergistically effective amount of such a second
immunosuppressive or anti-inflammatory agent.
The following examples are offered as a way for illustration of this invention and not in a
way of limitation.
Abbreviations used are:
Ac: Acetyl; Bz: Benzoyl; Bn: Benzyl; Ph: Phenyl; SEt: C2Hss; HRP: Horse radish peroxid-
ase; BSA: Bovine serum albumin; DMTST: Dimethyl(methylthio)sulfonium triflate; OTf: Tri-
flate; THG: Thioglycerol; THF: Tetrahydrofuran; NBA: m-Nitrobenzyl alcohol; DMF: N,N-Di-
methylformamide; DME: 1,2-Dimethoxyethane; MeOH: Methanol; PAA: Polyacryl amide;SA: Streptavidin; TBDMS: tert.butyldimethylsilyl; PTSA: p-toluene sulfonic acid; RT: room
CA 02260854 l999-0l-l2
W 098/06730 PCTAEP97/04279 -31-
temperature; MW: molecular weight; MS: mass spectroscopy; FAB: Fast atom bombard-
ment mass spe~lloscopy.
- An unconnected hyphen in the formulae means methyl.
A: Pl~p~ration of st~.li..~ cc~ ounds
E~ca,.~plt A1b: r~ o of cG,-",ound No. Alb
OH
'OOC~O",~ ~~
~ ~' O O
- J~,OH~NH + (Alb)
~J OH
HO
a) 2 (3.74 9 14 mmol)
HO"~
HO J t2)
O~BDMS
prepared according to Kinzy and Schmidt Tetrahedron LeK. 28:1981-1984 (1987) andimidate 3
OBz
BzO ~ I O~CCI3
BzO "~l NH (3)
BzO
[Rio et al. Carbohyd. Res. 219:71-90 (1991)] (12.76 9, 17 mmol) are dissolved in dry
CH2CI2 (100 ml) under argon at 0~C. 100 1ll (1.8 mmol) of a solution of triethylsilyltriflate
(117 ~LI in 2 ml CH2CI2) are added. After 4 h the reaction mixture is treated with NEt3 (0.5 ml)
and evaporated. Purification by repeated flash cl~r- i"atography on silica (ethyl acetate/-
hexane = 1/3) affords 4 as a white solid.
CA 02260854 1999-01-12
W O 98/06730 PCT~EP97/04279
-32-
OBz
O ' '~O HO X~~ (4)
BzO OTBDMS
b) 4 (740 mg, 0.88 mmol) is dissolved in 120 ml THF/pyridine 1/1 in a plastic container. At
0~C HF-pyridine complex 70130 (20 ml) is added. After 2 h sat. NaHCO3 solution is added
and the mixture is extracted with ethyl acetate. The organic layer is washed with sat. NH4CI,
dried with MgSO4 and evaporated. Purification by flash ~;I,rolnatography on silica ~ethyl
acetate/hexane = 1/1 ) gives 6 which is used directly in the next step.
OBz
BzO, ~ O " ~
BzO ' ~~ HO ~~ (6)
BzO OH
c) A solution of 6 (1.145 mg, 1.15 mmol), para-tolouenesulfonyl chloride (361 mg,
1.89 mmol) and dry pyridine (10 ml) in dry CH2CI2 (100 ml) is heated under reflux for 7 d.
The reaction mixture is evaporated and purified by flash chromatography on silica (ethyl
acetate/hexane = 2/3) to give in the order of elution 7 and recovered starting material.
OBz
BzO " ~ O ""~
BzO ~~ HO' Oo ~[~ (7)
BzO~ O ~'O
d) A solution of 7 (855 mg, 0.97 mmol) and NaN3 (253 mg, 3.89 mmol) in dry DMF (10 ml) is
stirred for 16 h. The reaction mixture is extracted with ethyl acetate, the organic layer is
washed with water, dried with MgSO4, evaporated and purified by flash chromatography on
silica (ethyl acetate/petrol ether = 1/2) to give 8.
OBz
N- (8)
CA 02260854 1999-01-12
W O 98/06730 PCTAEP97/04279
-33-
e) 8 (50 mg, 0.066 mmol) and imidate 9
J~
O CC13
O ~, OBn (9)
"' ~ OBn
BnO
[Wegmann and Schmidt, Carbohyd. Res. 184:254-261 (1988)] (58 mg, 0.100 mmol) aredissolved in ether (5 ml) under argon. 50 ~ul (0.05 eq.) of a solution of triethylsilyltriflate
(60 IlHn 4 ml ether) are added. After 15 min the reaction mixture is treated with NEt3, eva-
porated and purified by repeated flash chromatography on silica (ethyl acetate/hexane =
1/2) and subsequently by gel filtration on sephadex LH20 eluting with MeOH to give 10.
OBz
BzO, ~ ~~
BZO ~ - ~ N
BZO~ ~OBn N (10)
' ~-J OBn
BnO
f) A solution of 10 (160 mg, 0.137 mmol) in degassed MeOH (10 ml) is treated with a cata-
lytic amount of NaOMe solution. After 3 h the reaction mixture is neutralized with crushed
Amberlyst 15, filtered over hyflo, evaporated and purified by flash chromatography on silica
(chloroform/isopropanol = 8/1 ) to give 11.
OH
HO ' ~ " ~
HO : O
l~OBn N (11)
~ "' ~ OBn
BnO
.
CA 022608~4 1999-01-12
WO 98/06730 PCT~EP97/04279 -34-
g) 11 (1.17 9, 1.56 mmol) and dibutyltinoxide (Bu2SnO) (388 mg, 1.56 mmol) are suspended
in MeOH under argon and refluxed for 2 h. The resulting clear solution is evaporated. The
vacuum of the rotatory evaporator is rele~sed by flushing with argon not with air. The
residue is once evaporated with benzene and dried on high vacuum. Dried CsF (1.18 9,
7.8 mmol) is added under argon to the resulting residue and subsequently a solution of
benzyl-3-phenyl-2-trifluoromethanesulfonyloxypropionate 12
BnOOC~O~s,O
- '~ '''F
- O " 3 (12)
[Degerbeck et al., J. Chem. Soc. Perkin Trans. 1:11-14 (1993)] (3.0 9, 7.8 mmol) in DME
(50 ml). After stirring for 18 h at room temperature the mixture is diluted with ethyl acetate
and washed with sat. KH2PO4 and water, dried with MgSO4 and purified ~y flash chromato-
graphy on silica (ethyl acetate/hexane = 211 ) to give 13.
OH
BnOOC~O " ~ ~X~
~ ~,OBn N (13)
OBn
BnO
h) A mixture of t3 (902 mg, 0.91 mmol) dissolved in MeOH/H2O/AcOH 50150/1 (30.3 ml)
and Pd/C 10% (1 g) is stirred under H2 (balloon) for 50 h. Some ml of CH2CI2 are added.
The mixture is filtered over hyflo, evaporated, dissolved in water and freeze dried. The
resulting residue (5Z7 mg) is purified by P2 gelfiltration eluting with water to give 14.
OH
'OOCyO~ O~
~HO" ~ O
~J ~ J~, OH NH ~ (14)
' ~J OH
HO
CA 02260854 1999-01-12
WO 98/06730 PCT/EP97/04279
- 3~i -
i) A mixture of 14 (361 mg, 0.60 mmol) dissolved in H2O/dioxane 3/2 (45 ml) and
Rh 5%/AI2O3 (180 mg) is stirred under H2 (balloon) for 2 d. The mixture is filtered over hyflo,
evaporated, dissotved in water and purified by P2 gel~ alion eluting with water to give after
freeze drying pure Alb and an impure sample of A1b as white foams. C2,H47NO14
(MW=609.67): MS (FAB positive mode, THG) 632 (M+Na), 610 (M+H). 'H NMR (500 MHz,D2O) ~ 4.85 (d,1H, Fuc1), 4.62 (q,1H, Fuc5), 4.46 (d, 1H, Gal1), 4.05-3.96 (m, 2H), 3.94-
3.89 (m, 1 H), 3.85 (d,1 H, Gal4), 3.82 (dd, 111, Fuc3), 3.76-3.72 (m, 2H), 3.70-3.64 (m, 2H),
3.61-3.50 (m, 4H), 3.50-3.39 (m, 2H), 3.34 (dd,1H, Gal3), 3.14 (dd,1H), 2.21-2.16 (m, lH),
1.78-1.71 (m,1H),1.68-1.44 (m, 8H),1.22-1.08 (m, 6H, including at 1.15 (d, Fuc6)), 0.94-
0.80 (m, 2H).
Example A1c: Preparation of cG",~ound No. A1c
CO2Et
0~00~
BzO OBn OH (Alc)
H3Cf~ OBn
BnO
a) A mixture of 1c (1.00 g, 3.18 mmol)
HO~ SEt (1 c)
HO OBn
and dibutyltinoxide (1.38 g; 5.57 mmol,1.75 eq) in 50 ml of dry methanol is heated under
reflux for 2 h in an argon atmosphere. The solvent is removed and the residue dried in high
vacuo for 16 h. The colorless oil is dissolved in 50 ml of abs. dimethoxyethane. Under
argon, 2c (2.82 g, 9.54 mmol, 3.0 eq)
CO2Et
2-CF3 (2C)
~,1
-
CA 022608~4 1999-01-12
W 098/06730 PCT~EP97/04279 -36-
and dry cesiumfluoride (1.21 9, 7.95 mmol, 2.5 eq) are added and the resulting suspension
stirred at RT for 6 h. Then, 200 ml of a 1 N KH2PO4 solution containig 2 g of potassium-
fluoride is added followed by the extraction with chloroform (3 x 175 ml). The combined
organic layers are washed with brine (2 x 200 ml) and the solvent is removed. Flash
chromatography on silica gel (toluene/ethylacetate 5:1) gives 3c as a colorless oil.
CO2Et
[~HO OBn (3c)
b) To a solution of 3c (2.00 g, 4.03 mmol) in 18 ml of abs. pyridine benzoylchloride (2.8 ml,
24.1 mmol, 3.0 eq) and 4-(dimethylamino)-pyridine (0.147 g, 1.2 mmol, 0.3 eq) are added at
0~C. The mixture is stirred at RT for 16 h. Ethylacetate (300 ml) is added followed by the
extraction with 0.1 N HCI (5 x 100 ml), sat. NaHCO3 solution (5 x 100 ml) and brine (2 x
100 ml). The organic layer is dried with Na2SO4, the solvent is removed and the residue is
subjected to flash chromatography on silica gel (hexane/ethylacetate 4:1~2:1). Compound
4c is isolated as a colorless solid.
CO2Et
O BzO OBn (4c)
c) A suspension of 5c (4.00 g, 27.0 mmol)
HO~\
HO~ O (5c)
OH
6c (9.12 ml, 53.6 mmol)
OMe
MeO~< OMe (6c)
and PTSA (0.60 g) in 150 ml abs. acetonitrile is stirred at RT for 2 h. After the addition of
1.5 9 solid NaHCO3 the solvent is removed and the residue is subjected to flash chrc""ato-
graphy on silica gel (toluenelethylacetate 2:1~1:1). Compound 7c is isolated as a colorless
foam.
CA 022608~4 1999-01-12
W O 98/06730 PCT/EP97/04279
-37-
~~~ ~ O (7c)
d) A suspension of 4c (1.00 9, 1.42 mmol), 7c (0.756 g, 2.84 mmol, 2.0 eq) and molecular
sieves (3 A, 1.0 9) in abs. CH2C12 (5 ml) is stirred for 2 h under argon. Within 1 h a solution
of 8c (0.55 9, 2.13 mmol, 1.5 eq)
H3C
S--S--CH3 F3C-SO3 (8c)
_3
~Garegg, P.E., Carbohydrate Research 149:69 (1986)] in abs. CH2C12 (4 ml) is added drop-
wise at RT and the mixture is stirred for 1 h. Then, 50 ml of a sat. NaHCO3 solution are
added, the layers are separated and the aquous layer is extracted with CH2C12 (2 x 25 ml).
The combined organic layers are dried with Na2SO4, the solvent is removed and the residue
is dissolved in a mixture of CH2CI2 (50 ml) and methanol (10 ml). Camphersulfonic acid
(50 mg) is added and the mixture is stirred for 2 h at RT. The solvents are removed and the
residue is subjected to flash chromatography on silica gel (toluene/ethylacetate 2:1). Com-
pound 9c is isolated as a colorless oil.
CO2Et
~z~ OBn OH (9c)
e) A solution of 9c (847 mmg, 0.821 mmol) and triphenylmethyl chloride (343 mg, 1.23
mmol, 1.5 eq) in abs. pyridine (18 ml) is stirred under argon for 24h at 70~C. Then,
additional triphenylmethyl chloride (171 mg, 0.615 mmol, 0.75 eq) is added and stirring
continued for 24 h. The solvent is removed in vacuo and the residue purified by flash
chromatography on silica gel (toluene/ethylacetate 8:1~5:1) to yield 10c as a colorless
foam.
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
- 38 -
CO2Et
,~1 ~HO--~ (10c)
~ BzO OBn OC(Ph)3
f) A suspension of 10c (1000 mg, 0.969 mmol), tetraethylammonium bromide (405 mg,
1.94 mmol, 2.0 eq) and molecular sieves (3A, 1.2 9) in abs. CH2C12 (4.5 ml) and abs. DMF
(4.5 ml) is stirred under Argon for 2 h at RT (suspension A).
In a separate reaction flask a solution of bromine (357 mg, 2.23 mmol, 2.3 eq) in abs.
CH2CI2 is added at 0~C within 15 min to a solution of 11c (926 mg, 1.94 mmol, 2.0 eq),
SEt
H3C~OBn (11 C)
BnO
in abs. CH2C12 (1.5 ml). After stirring for 30 min cyclohexene (0.2 ml) is added and the
mixture is warmed to ~T (solution B).
The c~ear solution B is added dropwise to suspension A within 1 h. Having stirred for 2 h at
RT the mixture is diluted with ethylacetate (200 ml) and filtered through Hyflo Super Cel.
The solution is extracted with Na2S2O3 solution (100 ml), water (2 x 100 ml) and brine
(100 ml). The organic layer is concentrated and the residue dissolved in diethylether (25 ml)
and formic acid (5 ml). Having stirred for 3 h the solvents are removed and the residue is
purified by chromatography on silica gel (ethylacetate/hexane 2:1) to give A1c as a
colorless oil.1H NMR (400 MHz, CDCI3) ~ 0.50-1.42 (14 H, m, -CH2-cC6H1" H-2a,~), 1.17 (3
H, t, 7.0 Hz, CO2-CH2-CH3),1.34 (3 H, d, 6.5 Hz, H-6 Fuc), 1.94 (1 H, dd,13.015.0 Hz, H-
2eq)1 2.24 (1 H, t, 6.5 Hz, C6-OH), 3.16 (1 H, dt, 9.5/3.5 Hz, H-5), 3.29 (1 H, t (br), 12.0 Hz,
H-1 "~), 3.52 (1 H, d (br), 2.5 Hz, H-4 Fuc), 3.56 (1 H, dd,10.0/6.5 Hz, H-6 Gal), 3.57 (1 H, t,
9.5 Hz, H-4), 3.66 (1 H, dd,10.0/5.5 Hz, H-6' Gal), 3.74 (2 H, m, H-6, H-6'), 3.79 (1 H, dd,
12.0t5.0 Hz, H-1~ 3.84-3.89 (3 H, m, H-3, H-3 Gal, H-5 Gal), 3.92 (1 H, dd, 10.0/2.5 Hz, H-
3 Fuc), 4.05 (1 H, dd,10.0/3.5 Hz, H-2 Fuc), 4.07-4.15 (3 H, m, -CO2-CH2-CH3, -CH-CH2-
cC6H11),4.38 (1 H, d,11.5 Hz, -OCH2-Ph), 4.43 (1 H, d,11.5 Hz, -OCH2-Ph), 4.47 (1 H, d,
11.5 Hz, -OCH2-Ph), 4.48 (1 H, d, 11.5 Hz, -OCH2-Ph), 4.57 (1 H, d,11.5 Hz, -OCH2-Ph),
4.61 (1 H, q (br), 6.5 Hz, H-5 Fuc), 4.65 (1 H, d, 8.0 Hz, H-1 Gal), 4.68 (1 H, d,11.5 Hz, -
OCH2-Ph), 4.79 (1 H, d,11.5 Hz, -OCH2-Ph), 4.81 (1 H, d,11.5 Hz, -OCH2-Ph), 5.07 (1 H, d,
CA 02260854 1999-01-12
W 098/06730 PCT~EP97/04279 -39-
3.5 tlz, H-1 Fuc), 5.38 (1 H, dd,10.0/8.0 Hz, H-2 Gal), 5.89 (1 H, d, 3.5 Hz, H-4 Gal), 7.16-
8.12 (30 H, m, Ar-H); MS (FAB/EI) 1229 (M+Na)~.
B ~ zra~io~ of the ,~ t;cs
Example B(a): P~e pz..~t;G.~ of cG-,.~,ounds of the formula IE1~(A)
HO OH OH
R " O~b_O~_ORT'
NaO~O I HN~f~o (IEx(a))
H3C ~ OH R
r OH
OH
(1) Pre~a~alio,. of cG.,.pound No. B1a lR~: CH2C6Hll; RT': (CH2)8CO2CH3;
R~2: 3,4-(OCH3)2C6H3]
a) Within 1 h at -35~C in an argon atmosphere trimethylsilyl triflate (15.40 9; 69.50 mmol) is
added dropwise to a solution of tetraacetate 1a (12.00 g; 27.81 mmol)
OAc
AcO~OAc (1 a)
O ,~CH2
and 2a [HO(CH2)8CO2CH3] (7.84 g; 41.70 mmol) in abs. methylene chloride (150 ml). The
mixture is warmed to RT within 2 h and triethylamine (15 ml) are added. The mixture is
extracted successively with 0.1 n HCI, 0.1 n NaOH, water and saturated NaOH solution
(250 ml each). After filtration of the organic phase glycoside 3a is obtained by chromato-
graphy on silica gel (ether/hexane 3:1).
OAc
ACo~ - o(cH2)8co2cH3 (3a)
=~0 ~CH2
b) Triacetate 3a (10.40 9,18.60 mmol) is dissolved in abs. methanol (150 ml), mixed with
Amberlite IRA 910 in methanol (15 ml) and stirred at RT for 16 h. The mixture is filtered
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
- 40 -
through Hyflo Super Cel0, the solvent is removed and the residue is dried in vacuo to afford
glucosamine derivative 4a.
OH
H2O~o(cH2)8co2cH3 (4a)
=~0 ~CH2
c) Glucosamine derivative 4a (3.20 g; 7.39 mmol) is suspended in abs. acetonitrile (70 ml).
Benzaldehyde dimethylacetate (2.21 ml; 14.71 mmol) is added. Then p-toluenesulfonic acid
monohydrate (160 mg) is added and the mixture is stirred at Rt for 16 h and neutralized with
NaHCO3 (400 mg). The solvent is removed in vacuo. Chrumatography on silica gel (chloro-
form/acetone 10:1 ) affords the partially protected carbohydrate 5a.
Ph ~~O~~_ OtCH2)8cO2cH3 (5a)
=~o ~ CH2
d) Glucosamine derivative 5a (452 mg; 0.870 mmol), tetraethylammonium bromide (400 mg;
1.90 mmol) and 1.0 g activated molecular sieves (4 A) are suspended in a mixture of abs.
methylene chloride (6.0 ml) and abs. N,N-dimethylformamide (4.3 ml) and stirred in an
argon atmosphere at RT for 1 h (suspension A).
In a separate reaction vessel a solution of bromine in abs. methylene chloride (0.10 ml in
0.5 ml) is added (argon atmosphere; 0~C) to a solution of fucose derivative 11c (830 mg;
1.74 mmol) in a~s. methylene chloride (2.5 ml) within 15 min. Having stirred for 30 min
cyclohexene (0.25 ml) is added and the mixture is warmed to RT (solution B).
The clear solution B is added dropwise to suspension A within 1 h. Having stirred for 16 h
the mixture is diluted with ethyl acetate (50 ml) and filtered through Hyflo Super Cel~. The
solution is successively extracted with sodium thiosulfate solution (50 ml), twice with water
(50 ml each) and saturated NaCI solution (50 ml each). The organic phase is dried over
sodium sulfate and concentrated in vacuo. Chromatography on silica gel (hexane/ethyl
acetate 2:1 ) affords disaccharide 7a.
CA 02260854 1999-01-12
WO 98/06730 PCT/EP97/04279
- 41 -
Ph~O'~_O(C~)bCO2C,~,
o--'O ~CH2 (7a)
H3C~_OBn
OBn OBn
e) Disaccharide 7a (1.00 9; 1.07 mmol), sodium cyano borohydride (670 mg; 10.70 mmol)
and 2.0 g activated molecular sieves (3 A) are suspended in abs. THF (20 ml). At 0~C a
saturated solution of HCI gas in abs. ether is added dropwise. Before the complete con-
sumption of 7a ethyl acetate (50 ml) is added and the solution is filtered through Hyflo
Super Cel~. The solution is successively extracted twice with NaHCO3 solution (50 ml each),
water (50 ml each) and saturated NaCI solution (50 ml each). The organic phase is dried
over Na2SO4 and concentrated in vacuo. Chromatography on silica gel (hexane/ethyl
acetate 2:1) affords disaccharide 8a.
OBn
HO~O(C~ co2cH3 (8a)
H3C~;~OBn
OBn
f) Disaccharide 8a (460 mg, 490 mmol) and morpholine (1400 mg, 16.0 mmol) are dissolved
in abs. THF (10 ml) (RT; argon atmosphere). Pd[P(Phh]4 (58 mg, 0.050 mmol) is added and
the solution is stirred for 15 min at RT. The volatile components are removed in vacuo.
Chromatography on silica gel (ethyl acetate/hexane 1:1~2:1) affords amino sugar 9a.
OBn
HO ~_ O(CH2)3C~2C~3
NH2 (9a)
H3C~,OBn
OBn OBn
g) Amino sugar 9a (370 mg, 0.433 mmol) and active ester 1 Oa (206 mg, 0.591 mmol)
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97/04279
F5
@~
~ (1 Oa)
o~H3
OCH3
are dissolved in abs. N,N-dimethylformamide (3.5 ml) (argon atmosphere). 2,6-Lutidine
(0.7 ml) is added and the solution is warmed to 70~C for 4 h. Another 50 mg (0.143 mmol)
of 1 Oa are added and the solution is stirred for 16 h at 70~C. Then another 30 mg
(0.086 mmol) of 10a are added and the solution is warmed to 70~C for 3 h. Ethyl acetate
(50 ml) is added and the solution is successively extracted twice with ammonium sulfate
solution (50 ml each), NaHCO3 solution (50 ml each) and saturated NaCI solution (50 ml
each). After filtration through cotton wool the solvent is evaporated in vacuo. Chromato-
graphy of the residue on silica gel (chloroform/ethyl acetate 3:1) affords amide 11a.
OBn
HO ~ O(CH2)8C02cH3
H3C~OBn ~OCH3 (11a)
h) Disaccharide 11a (190 mg, 0.186 mmol), thioglycoside 12a (175 mg, 0.279 mmol)
~zO OBn
BzO~SEt (12a)
and activated molecular sieves (4 A)(500 mg) are suspended in 1.5 ml abs. methylene
chloride and stirred in an argon atmosphere for 1 h at RT (solution A).
In a separate reaction vessel activated molecular sieves (3 A)(500 mg) are added to a solu-
tion of dimethyl(methylthio)sulfonium triflate 8c (144 mg, 0.558 mmol) in abs. methylene
chloride (1.5 ml) and the mixture is stirred for 1 h at RT in an argon stmosphere (solution B).
Solution B is added dropwise to solution A within 4 h. Then the mixture is diluted with ethyl
acetate (50 ml), filtered through Hyflo Super Cel~ and successively extracted with NaHCO3
solution (50 ml) and twice with saturated NaCI solution (25 ml each). The organic phase is
dried over sodium sulfate and concentrated. Chromatography on silica gel (chlorofor",e/
ethyl acetate 6:1) affords trisaccharide 14a.
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97/04279
-43-
BzO OBn OBn
BzO ~CH3
i) Trisaccharide 14a (205 mg, 0.129 mmol) is dissolved in abs. methanol (5 ml) in an argon
atmosphere. Then 0.065 ml of a 2.0 M solution of sodium methanolate (0.13 mmol) in abs.
methanol are added and the solution is stirred for 3 h at RT. The solution is neutralized by
the addition of acetic acid (0.01 ml~ and the solvent is removed in vacuo.Chromatography
on silica gel (chloroform/acetone 2:1) affords partially deprotected trisaccharide 15a.
HO OBn OBn
(15a)
OBn
j) Trisaccharide 15a (80 mg, 0.063 mmol) is dissolved in an argon atmosphere in abs. ben-
zene (2 ml), dibutyltinoxide (28 mg, 0.110 mmol) is added and the mixture is heated under
reflux for 16 h. The solvent is removed, the residue is dried for 1 h at 40~C in high vacuum
and dissolved in abs. dimethoxyethane (1.6 ml). A solution of triflate 16a (125 mg,
0.31 5 mmol)
o
L~OSO2CF3 (16a)
BrlO O
in abs. dimethoxyethane (1 ml) is added and the mixture is stirred for 6 h at 40~C. Then
25 ml KH2PO4/potassium fluoride solution is added and the solution is extracted twice with
chloroform (25 ml each). The solution is dried with sodium sulfate and the solvent is re-
moved. Chromatography on silica gel (chloroform/acetone 15:1) affords trisaccharide 17a.
CA 02260854 1999-01-12
W O 98/06730 PCTAEP97/04279
-44-
HO OBn OBn
HO O=~O(CH~)bC~CH3 (17a)
B~OBn ~OCH,
k) Trisaccharide 17a (44 mg, 29 ~Imol) is dissolved in a mixture of dioxane (4 ml), water
(1.5 ml) and acetic acid (0.25 ml). Having thoroughly degassed p~ umhydroxide (20%)
on coal (100 mg) is added. The suspension is stirred for 15 min in an argon atmosphere.
Then tl~e mixture is hydrogenated for 16 h at RT. The catalyst is filtered off through a HPLC
filter, the solvent is removed, the residue is dissolved in water/methanol (2:1) and passed
through a sodium ion exchange column (water). Product containing fractions are combined
and concentrated. The residue is dissolved in water and Iyophilized to obtain compound
Bla.1H-NMR (400 MHz, D20) â 0.50-1.70 (25 H, m, -O-CH2(CH2)6CHzCO2CH3, -CH;~c-
C6H"~,1.07 (3 H, d, 6.5 Hz, 3 x H-6 Fuc), 2.06 (2 H, t, 7.5 Hz, -CH2CH2CO2Me), 3.30 (1 H,
dd, 9.5/3.0 Hz), 3.42-4.15 (17 H, m), 3.54 (3 H, s, -CO2CH3), 3.77 (3 H, s, Ar-OCH3), 3.78 (3
H, s, Ar-OCH3), 4.37 (1 H, d, 8.0 Hz, H-1 Gal), 4.55 (1 H, s (br), H-1 Glc), 4.71 (1 H, q, 6.5
Hz, H-5 Fuc), 5.02 (1 H, d, 3.5 Hz, H-1 Fuc), 7.01 (1 H, d, 8.5 Hz, Ar~, 7.32 (1 H, d, 2.0 Hz,
ArH), 7.38 (1 H, dd, 8.5/2.0 Hz, Ar~; MS (FAB/EI) 974 (M - Na).
(2) Preparation of compound No. B2a [R4: CH2C6H"; RT': (CH2)~CO2CH3;
RT2: 3,4-(OH)2C6H3]
Starting with amino sugar 9a (150 mg, 0.175 mmol) and active ester 28a (175 mg,
0.351 mmol) F5
~'
~ (28a)
O~OBn
OBn
compound No. B2a is prepared according to Example B(a)(1). 'H-NMR (400 MHz, D2O) ~
0.60-1.73 (25 H, m, -O-CH2(CH2)6CH2CO2CH3, -CH2-c-C6H11),1.07 (3 Il, d, 6.5 Hz, 3 x H-6
~uc), 2.13 (2 H, t, 7.5 Hz, -CH2CH2CO2Me), 3.32 (1 H, dd, 9.513.0 Hz), 3.44-3.95 (17 H, m),
3.57 (3 H, s, -CO2CH3), 4.39 (1 H, d, 8.0 Hz, H-1 Gal), 4.56 (1 H, s (br), H-1 Glc), 4.71 (1 H,
CA 02260854 1999-01-12
W 098/06730 PCT/EP97/04279 -45-
q, 6.5 Hz, H-5 Fuc), 5.02 (1 H, d, 3.5 Hz, H-1 Fuc), 6.87 (1 H, d, 8.5 Hz, ArH), 7~21 (1 H, dd,
8.5/2.0 Hz, ArH), 7.24 (1 H, d, 2.0 Hz, ArH); MS (FAB/EI) 946 (M - H)-.
(3~ Preparation of cG,.,pound No. B3a [R4: C~H11; RT': (CH2)8CO2CH3;
RT2: 3,4-(OCtl3)2C6H3]
a) Starting from trisaccharide 15a (77 mg, 0.061 mmol) and triflate 23a (120 mg,0.303 mmol)
C~OSO2CF3 (23a)
BnO O
compound No. B3a is prepared accGI~Jing to Example B(a)(1)- 'H-NMR (400 MHz, D20) ~
0.50-1.63 (23 H, m, -O-CH2(CH2)6CH2CO2CH3, c-C6H,1), 1.06 (3 H, d, 6.5 Hz, 3 x H-6 Fuc),
2 04 (2 H, t, 7.5 Hz, -CH2CH2CO2Me), 3.26 (1 H, dd, 9.5/3.0 Hz), 3.39-3.92 (17 H, m), 3.54
(3 H, s, -CO2CH3), 3.77 (3 H, s, Ar-OCH3), 3.78 (3 H, s, Ar-OCH3), 4.36 (1 H, d, 8.0 Hz, H-1
Gal), 4.55 (1 H, s (br), H-1 Glc), 4.71 (1 H, q, 6.5 Hz, H-5 Fuc), 5.00 (1 H, d, 3.5 Hz, H-1
Fuc), 6.98 (1 H, d, 8.5 tlz, ArH), 7.31 (1 H, d, 2.0 Hz, ArH), 7.37 (1 H, dd, 8.5/2.0 Hz, ArH);
MS (FAB/EI) 960 (M - Na)~.
(4) Preparation of compound No. B4a [R4: CH2C6H"; RT': (CH2)~CO2Na;
RT2 3 ,4 (OCH3)2C6H3]
Methyl ester Bla (10 mg, 0.010 mmol) is dissolved in water (1 ml), mixed with 2 N NaOH
(20 ~I) and stirred for 16 h at RT. Reverse phase chromatoglaphy on RP 18 (water~
water/methanol 3:1) affords carboxylate B4a. 'H-NMR (400 MHz, DzO) ~ 0.49-1.70 (25 H,
m, -O-CH2(CH2)6CH2CO2CH3, -CH~c-C6H11), 1.06 (3 H, d, 6.5 Hz, 3 x H-6 Fuc), 1.92 (2 H, t
(br), 7.5 Hz, -CH2CH2CO2Me), 3.28 (1 H, dd, 9.5/3.0 Hz), 3.42-4.15 (17 H, m), 3.77 (3 H, s,
Ar-OCH3), 3.78 (3 H, s, Ar-OCH3), 4.36 (1 H, d, 8.0 Hz, H-1 Gal), 4.53 (1 H, s (br), H-1 Glc),
4.71 (1 H, q, 6.5 Hz, H-5 Fuc), 5.02 (1 H, d, 3.5 Hz, H-1 Fuc), 7.00 (1 H, d, 8.5 Hz, ArH),
7.31 (1 H, d, 2.0 Hz, ArH), 7.37 (1 H, dd, 8.5/2.0 Hz, Art~; MS (FAB/EI) 1004 (M - H)-.
CA 022608~4 1999-01-12
W O 98/06730 PCTAEP97/04279
-46-
Example B(b): Pl~, z.dl;ol~ of cG.,.~ounds of the formula IEXIb
OH
NaOOCrO "~ ~'~"~
~HO'I--O O~O
'J HO o~OH NR~6RB' (IEX(b))
H3C OH
HO
(1) P~ ardlion of c~ ound No. Blb ~R~6: H, R~': C(O)CH(C6H5)2~
A solution ot commercially available diphenylacetyl chloride (11.3 mg, 0.049 mmol, 1.5 eq.)
in THF (0.5 ml) is added at 0~C to a solution of A1b (20 mg, 0.033 mmol) in THF/H2O 1/1
(2 ml). The pH of the reaction mixture is adjusted to 8-10 by the addition of 1 N NaOH solu-
tion and maintained at 8-10 throughout the whole reaction. After 18 h additional diphenyl-
acetyl chloride (3.7 mg, 0.016 mmol, 0.5 eq.) is added and after a total of 42 h the reaction
mixture is partially evaporated to remove THF. The now aqueous solution is purified by RP
C18 (column size 1 x 10 cm) through stepwise elution with acetonitrile/water 30/70 and then
acetonitrile/water 40t60. The product obtained is further purified by flash chromatography
on silica (ethyl acetate/isopropanol/water = 4/2/1) to give after freeze drying B1 b as a white
foam. C41Hs6No1sNa (MW=825.88): MS (FAB positive mode, THG) 826 (M+H), 804 (M-
Na+H).1H NMR (500 MHz, D2O) ~ 7.42-7.22 (m, 20H), 5.16 (s, 1H, CHPh2), 4.78 (d, lH,
Fuc1), 4.64 (q, 1 H, Fuc5), 4.46 (d,1 H, Gal1), 4.02-3.88 (m, 3H),3.87 (d,1 H, Gal4), 3.82
(dd,1H, Fuc3), 3.78-3.65 (m, 5H), 3.62-3.49 (m, 3H), 3.45-3.32 (m, 4H), 2.19-2.11 (m, lH),
1.79-1.72 (m,1H),1.68-1.46 (m, 8H),1.22-1.08 (m, 6H, including at 1.15 (d, Fuc6)), 0.96-
0.82 (m, 2H).
The following compounds are prepared in analogy to the above example whereas for the
purification one to three of the following purification steps are applied in any order desired
to obtain analytically pure compounds:
a) Reverse phase C18 column chromatography (column size 1 x 10 cm) using stepwise
elution with acetonitrile/water starting with low acetonitrile content ending with high
acetonitrile content.
b) Flash chromatography on silica (ethyl acetate/isopropanol/water = 41211)
c) P2 gelfiltration using water as eluent.
CA 022608~4 1999-01-12
W O 98/06730 PCTrEPg7/04~79
- 47 -
(2) P~ dlion of cG.,.?ound No. B2b lRB6: H, RB': C(O)C6H"]
C34H~6NO15Na (MW=741.80): MS (FAB positive mode, THG) 742 (M~H), 720 (M-Na+H).1HNMR (500 MHz, D2O) ~ 4.86 (d, 1 H, Fuc1), 4.67 (q, 1 H, Fuc5), 4.48 (d, 1 H, Gall ), 4.05-3.91
(m, 3H), 3.87 (d, 1H, Ga.4), 3.83 (dd, 1H, Fuc3), 3.77 (d, 1H, Fuc4), 3.74 (dd, 1H, Fuc2),
3.72-3.68 (m, 2H), 3.63-3.55 (m, 3H), 3.53-3.40 (m, 4H), 3.36 (dd, 1H, Gal3), 2.27-2.16 (m,
2H), 1.79-1.68 (m, 5H), 1.68-1.47 (m, 9H), 1.37-1.08 (m, 11H, including at 1.17 (d, Fuc6)),
0.97-0.83 (m, 2H).
(3) ~ rdticl) of col..pound No. B3b tR~6: H, R~': C(O)C6H53
C34Hs0No~sNa (MW=735.76): MS (FAB negative mode, THG) 712 (M-Na). 'H NMR (400
MHz, D2O) ~ 7.66 (d, 2H), 7.52 (m, 1 H), 7.42 (t, 2H), 4.93 (d, 1 H, Fuc1), 4.64 (q, 1 H, Fuc5),
4.42 (d, 1 H, Gal1), 4.04-3.90 (m, 3H), 3.88-3.75 (m, 3H including: 3.85 (d, 1 H, Gal4), 3.80
(dd, 1 H, Fuc3)), 3.75-3.68 (m, 2H, Fuc4, Fuc2), 3.69-3.36 (m, 8H), 3.32 (dd, 1 H, Gal3),
2.1 8-2.10 (m, 1 H), 1.73-1.65 (m, 1 H), 1.65-1.39 (m, 8H), 1.21-1.00 (m, 6H, including at 1.13
(d, Fuc6)), 0.92-0.73 (m, 2H).
(4) Preparation of co.,.~ound No. B4b ERR6: H, R~7: C(O)C6H4(4-OCH3)]
C3sHs2No16Na (MW=765.78): MS (FAB negative mode, THG) 742 (M-Na). 'H NMR (400
MHz, D2O) ~ 7.66 (m, 2H), 6.98 (m, 2H), 4.92 (d, 1 H, Fuc1), 4.63 (q, 1 H, Fuc5), 4.42 (d, 1 H,
Gall), 4.04-3.85 (m, 3H), 3.83-3.75 (m, 6H including: 3.81 (d, 1H, Gal4), 3.79 (s, 3H,
OCH3), 3.78 (dd, 1 H, Fuc3)), 3.73-3.68 (m, 2H, Fuc4, Fuc2), 3.68-3.35 (m, 8H), 3.30 (dd,
1H, Gal3), 2.18-2.11 (m, 1H), 1.73-1.65 (m, 1H), 1.65-1.39 (m, 8H), 1.21-1.00 (m, 6H,
including: 1.10 (d, Fuc6)), 0.92-0.75 (m, 2H).
(5) ~ ardlion of cGl"pound No. B5b [RB6: H, R~': C(O)C6H3(3,4-OCH3)21
C36Hs4NOl7Na (MW=795.81): MS (FAB negative mode, THG) 794 (M-H), 772 (M-Na). lH
NMR (500 MHz, D2O) ~ 7.40 (dd, 1 H), 7.35 (d, 1 H), 7.06 (d, 1 H), 4.98 (d, 1 H, Fuc1), 4.69 (q,
1 H, Fuc5), 4.49 (d, 1 H, Gal1), 4.05 (m, 1 H), 3.99 (m, 1 H), 3.93 (m, 1 H), 3.90-3.81 (m, 9H
including: 3.87 (s, 3H, OCH3), 3.86 (s, 3H, OCH3), 3.75 (dd, 1H, Fuc3)), 3.80-3.75 (m, 2H,
Fuc4, Fuc2), 3.74-3.54 (m, 6H), 3.54-3.44 (m, 2H), 3.36 (dd, 1H, Gal3), 2.24-2.19 (m, 1H),
1.78-1.72 (m, 1 H), 1.69-1.45 (m, 8H), 1.24-1.08 (m, 6H, including: 1.17 (d, Fuc6)), 0.96-0.82
(m, 2H).
_ .. , . .. , ,. ~
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
- 48 -
(6) P~pardtion of cG."pound No. B6b lR86: H, R~': C(O)C6H4(4-CI)]
C34H49NO,5NaCI (MW=770.20): MS (FAB negative mode, THG) 768 (M-H), 746 (M-Na).1HNMR (400 MHz, D20) ~ 7.62 (m, 2H), 7.42 (m, 2H), 4.92 (d, 1 H, Fuc1), 4.63 (q, 1 H, Fuc5),
4.42 (d, 1 H, Gal1), 4.04-3.84 (m, 3H), 3.83-3.75 (m, 3H including: 3.81 (d, 1 H, Gal4), 3.78
(dd, 1 H, Fuc3)), 3.73-3.68 (m, 2H, Fuc4, Fuc2), 3.68-3.35 (m, 8H), 3.30 (dd, 1 H, Gal3),
2.18-2.10 (m, 1H), 1.73-1.65 (m, lH), 1.65-1.37 (m, 8H), 1.21-1.00 (m, 6H, including: 1.10
(d, Fuc6)), 0.92-0.72 (m, 2H).
(7) Ple:pardlion of cG"".ound No. B7b lRB6: H, RB7: C(O)C6H4(4-NO2)]
C34H49N20,7Na (MW=780.75): MS (FAB negative mode, THG) 779 (M-H), 757 (M-Na). lHNMR (400 MHz, D20) ~ 8.25 (m, 2H), 7.83 (m, 2H), 4.92 (d, 1 H, Fuc1), 4.63 (q, 1 H, Fuc5),
4 42 (d, 1 H, Gal1), 4.04-3.84 (m, 4H), 3.83-3.76 (m, 2H, Gal4, Fuc3), 3.74-3.58 (m, 5H),
3.68-3.35 (m, 5H), 3.29 (dd, 1H, Gal3), 2.18-2.08 (m, 1H), 1.72-1.64 (m, lH), 1.64-1.36 (m,
8H), 1.21-0.96 (m, 6H, including: 1.10 (d, Fuc6)), 0.90-0.75 (m, 2H).
(8) Preparation of cG-,-pound No. B8b lR~6: H, RB': C(O)C6H4(4-C6H5)]
C40HssNo1sNa (MW=812.86): MS (FAB positive mode, THG) 790 (M-Na~H). 'H NMR (400
MHz, D20) ~ 7.78 (d, 2H), 7.75-7.65 (m, 4H), 7.48 (t, 2H), 7.42 (m, 1 H), 4.97 (d, 1 H, Fuc1),
4.66 (q, 1 H, Fuc5), 4.45 (d, 1 H, Gal1), 4.08-3.80 (m, 6H), 3.80-3.72 (m, 2H, Fuc4, Fuc2),
3.72-3.62 (m, 3H), 3.62-3.38 (m, 5H), 3.32 (dd, 1H, Gal3), 2.21-2.13 (m, 1H), 1.76-1.68 (m,
1H), 1.68-1.40 (m, 8H), 1.22-1.00 (m, 6H, including: 1.13 (d, Fuc6)), 0.95-0.78 (m, 2H).
(9) Preparation of compound No. B9b [RB6: ti, RB7: C(0)-2-na"l.ll.~l]
C3BHs2NO1sNa (MW=785.82): MS (FAB negative mode, THG) 762 (M-Na). 'H NMR (500
MHz, D20) ~ 8.28 ~s, 1 H), 8.02-7.93 (m, 3H), 7.78-7.74 (m, 1 H), 7.65-7.58 (m, 2H), 5.01 (d,
1 H, Fuc1), 4.70 (q, 1 H, Fuc5), 4.49 (d, 1 H, Gal1), 4.01-3.97 (m, 2H), 3.96-3.85 (m, 4H
including Gal4, Fuc3), 3.81-3.76 (m, 2H, Fuc4, Fuc2), 3.76-3.68 (m, 3H), 3.65-3.45 (m, 5H),
3.36 (dd, 1H, Gal3), 2.24-2.18 (m, 1H), 1.79-1.72 (m, 1H), 1.72-1.46 (m, 8H), 1.24-1.06 (m,
6H, including: 1.10 (d, Fuc6)), 1.00-0.81 (m, 2H).
(10) Preparation of compound No. BlOb [R~6: H, R~7: C(O)OCH2C6Hs]
C3sHs2No16Na (MW=765.82): MS (FAB negative mode, THG) 742 ~M-Na). 'H NMR (500
MHz, D20) ~ 7.42-7.33 (m, 5H), 5.06 (d, 2H, CH2Ph), 4.86 (d, 1 H, Fuc1), 4.66 (q, 1 H, Fuc5),
CA 022608~4 1999-01-12
W O 98/06730 PCT/EP97/04279
-49-
4.45 (d, 1 H, Gal1), 4.01 -3.95 (m, 1 H), 3.95-3.89 (m, 2H), 3.85 (d, 1 H, Gal4), 3.81 (dd, 1 H,
Fuc3), 3.76-3.66 (m, 4H), 3.62-3.54 (m, 3H), 3.44-3.31 (m, 5H), 2.18-2.12 (m, 1H), 1.76-
1.71 (m, 1H), 1.65-1.44 (m, 8H), 1.21-1.03 (m, 6H, including: 1.13 (d, Fuc6)), 0.94-0.80 (m,
2H).
(11) rl qJ~.d~iol~ of co~ ,ound No. B11b [RBs: H, RB7: C(O)NHCsH5]
A solution of phenylisocyanate (4 mg, 0.033 mmol, 1.2 eq.) in 0.5 N NaOH is added at 0~C
to a solution of A1b (17 mg, 0.027 mmol) in 0.5 N NaOH (1 ml). Addition of phenylisocyan-
ate is continued until after 11 d a total amount of 6 equivalents is added. The product is
purified by flash chromatography on silica (ethyl acetate/isopropanol/water = 41211), filtered
on Dowex ion exchange resin Na~ form eluting with water, further purified by P2 gelfiltration
using water as eluent and again filtered on Dowex Nat form to give after freeze drying B11 b
as a white foam. C34Hs,N2O,sNa (MW=750.77): 1H NMR (400 MHz, D2O) ~ 7.27 (m, 2H),
7.18 (d, 2H), 7.03 (t, 1H), 4.86 (d, lH, Fuc1), 4.64 (q, 1H, Fuc5), 4.39 (d, lH, Gal1), 4.08-
3.85 (m, 3H), 3.84 (s, 1H, Gal4), 3.76 (dd, 1H, Fuc3), 3.72-3.64 (m, 2H, Fuc4, Fuc2), 3.63-
3.46 (m, 6H), 3.46-3.28 (m, 4H), 2.16-2.06 (m, lH), 1.73-1.63 (m, 1H), 1.72-1.38 (m, 8H),
1.20-0.97 (m, 6H, including: 1.09 (d, Fuc6)), 0.92-0.72 (m, 2H).
(12) Preparation ot compound No. B12b [RBs: H, RB': SO3Na]
Commercially available sulfur trioxide pyridine complex (7.8 mg, 0.049 mmol, 1.5 eq.) is
added to a solution of Alb (20 mg, 0.033 mmol) in H20 (2 ml) with enough 2 N NaOH to
obtain a pH of above 11. After 16 h another portion of sulfur trioxide pyridine complex
(7.8 mg, 0.049 mmol, 1.5 eq.) is added. After 40 h the reaction mixture is evaporated and
purified by P2 gelfiltration eluting with water and the product is subsequently purified by C18
reverse phase chromatography (using a SepPac syringe adapter) through stepwise elution
with acetonitrile/water 10/90 ~ 90/10 to give after evaporation and freeze drying B12b as a
white foam. C27H4sNO,7SNa2 (MW=733.70): MS (FAB negative mode, THG) 710 (M-Na),
688 (M-2Na+H). 'H NMR (400 MHz, D2O) ~ 4.98 (d, 1 H, Fuc1), 4.68 (q, 1 H, Fuc5), 4.42 (d,
1 H, Gal1), 3.99-3.86 (m, 3H), 3.82 (d, 1 H, Gal4), 3.78 (dd, 1 H, Fuc3), 3.73-3.67 (m, 2H,
Fuc4, Fuc2), 3.67-3.59 (m, 2H), 3.58-3.48 (m, 2H), 3.48-3.32 (m, 4H), 3.30 (dd, 1 H, Gal3),
3.12 (dd, lH), 2.17-2.08 (m, lH), 1.74-1.66 (m, 1H), 1.63-1.39 (m, 8H), 1.21-1.00 (m, 6H,
including at 1.12 (d, Fuc6)), 0.92-0.75 (m, 2H).
CA 022608~4 1999-01-12
W O 98/06730 PCT~EP97/04279
-50-
(13) P,~a(d~ion of co,..pound No. B13b lRB6: H, RB7: CH2C6H5l and cc...,uound No.
B14b [RB6: CH2C6H5, R~7: CH2C6H5l
Borane pyridine complex (BH3.C5H5N, 0.013 ml, 0.131 mmol) is added to a mixture of Alb
(40 mg, 0.066 mmol), benzaldehyde (0.033 ml, 0.328 mmol) and freshly dried 4A mol~cu! r
sieves (ca. 500 mg) in dry MeOH (0.5 ml). After 20 h two new products have formed and the
reaction mixture is filtered, evaporated and the two products are separated by flash
chromatography on silica (ethyl acetate/isopropanol/water = 412/1) to give in the order of
elution fraction 1 and fraction 2. Fraction 1 is further purified by P2 gelfiltration eluting with
water and then ~iltered on Dowex ion exchange resin Na~ form eluting with water to give
after freeze drying B14b as a white foam. Fraction 2 is also further purified by P2 gelfiltra-
tion eluting with water and then filtered on Dowex ion exchange resin Na+ form eluting with
water to give after freeze drying B1 3b as a white foam.
B14b: C41H5HNOl4Na (MW=811.91): MS (FAB negative mode, THG) 788 (M-Na). 'H NMR
(400 MHz, D2O) ~ 7.41 (m, 10H), 4.64 (d, 1H, Fuc1), 4.43 (q, 1H, Fuc5), 4.37 (d, 1H, Gal1),
4.46-4.12 (m, 3H), 3.93-3.83 (m, 2H), 3.81 (s, 1H Gal4), 3.78-3.58 (m, 7H), 3.58-3.37 (m,
3H), 3.37-3.19 (m, 3H), 2.12-2.02 (m, lH), 1.74-1.65 (m, 1H), 1.73-1.38 (m, 8H), 1.22-0.97
(m, 6H, including: 1.08 (d, Fuc6)), 0.92-0.75 (m, 2H).
B13b: C34Hs2NOl4Na (MW=721.77): MS (FAB negative mode, THG) 698 (M-Na). lH NMR
(400 MHz, D2O) ~ 7.38 (m, 5H), 4.72 (d, 1 H, Fuc1), 4.52 (q, 1 H, Fuc5), 4.41 (d, 1 H, Gal1),
4.19 (d, 1H, CH2Ph), 4.14 (d, 1H, CH2Ph), 4.00-3.84 (m, 3H), 3.81 (s, 1H, Gal4), 3.73 (dd,
1 H, Fuc3), 3.69 (d, 1 H), 3.68-3.59 (m, 3H), 3.59-3.45 (m, 4H), 3.44-3.32 (m, 2H), 3.29 (dd,
lH, Gal3), 3.18 (dd, 1H), 2.18-2.08 (m, 1H), 1.73-1.65 (m, 1H), 1.72-1.38 (m, 8H), 1.21-0.95
(m, 6H, including: 1.09 (d, Fuc6)), 0.92-0.72 (m, 2H).
The following compounds are prepared in analogy to the above examples:
Compound No. R~6 R~'
B15b H C(O)CH2C6Hs
B16b H C(O)(CH2)2C6Hs
B17b H C(O)C6H4(4-CF3)
B18b H C(O)COONa
B19b H C(0)-1 -naphthyl
B20b H C(O)NHCH2CH3
CA 02260854 1999-01-12
WO 98/06730 PCT/EP97/04279
- 51 -
Compound No. R~6 R~'
B21 b H C(O)NH(CH2)2COONa
B22 HC(O)OCH2C6H4(4-NO2)
B23b tlC(O)OCH2C6H2(2-NO2)(4.5-OCH3)2
B24b H C(O)OCH2-2-naphthyl
B25b H CH3
B26b H SO2C6H4(4-CH3)
B27b CH3 C(O)c6Hs
B28b CH2C6H5 C(O)C6H6
ExamP~e B(c)~ ardtiGn of cG.~ ounds of the formula IE,~(C)
CO2Na
~HO~O~RS (IEX~C))
H3CI~ ~H
(1) Preparation of cG,."~ound No. B1c [Rs: CH2OH~
To a solution of A1c (75 mg, 0.062 mmol) in dioxane (1 ml) 0.5 ml of 0.5 N NaOH are added
and the mixture is stirred at RT for 1 h. Then, 0.05 ml of acetic acid are added and the sol-
vent is removed in vacuo. The residue is dissolved in abs. methanol (1 ml) and hydrogena-
ted in the presence of Pd(10%) on charcoal (75 mg) for 16 h at RT. The catalyst is filtered
off, the solvent is removed and the residue is purified by reversed phase chromatography
(RP18, water/methanol 3:1~1:2). Passage through a sodium ion exchange column follow-
ed ~y Iyophilization gives ~1c as a colorless powder. lH NMR (400 MHz, D2O) ~ 0.75-1.72
(13 H, m, -CH7-cC6_11, H-2",~), 1.09 (3 H, d, 6.5 Hz, H-6 Fuc), 2.12 (1 H, dd, 13.0/5.0 Hz, H-
2eq)~ 3.28 (2 H, m, H-5, H-3 Gal), 3.38 (1 H, t (br), 12.5 Hz, H-1ax)~ 3.47 (1 H, t, g.5 Hz, H-4),
3.50 (1 H, m, H-5 Gal), 3.52 (1 H, t, 8.5 Hz, H-2 Gal), 3.62 (2 H, m, H-6 Gal, H-6' Gal), 3.67
(1 H, dd, 10.5/4.0 Hz, H-2 Fuc), 3.69 (1 H, d (br), 3.0 Hz, H-4 Fuc), 3.74 (1 H, dd, 7.0/4.0
Hz, H-6), 3.78 (1 H, dd, 10.0/3.0 Hz, H-3 Fuc), 3.80 (1 H, d (br), 3.0 Hz, H-4 Gal), 3.84 (1 H,
dd, 7.0/2.0 Hz, H-6'), 3.88 (2 H, m, H-1eq, -C_-CH2-cC6H"), 3.94 (1 H, td, 9.5/5.0 Hz, H-3),
.
CA 02260854 l999-0l-l2
W O 98/06730 PCT/EP97/04279
-52-
4.41 (1 H, d, 8,5 Hz, H-1 Gal), 4.71 (1 ~, 4 (br), 6.5 Hz, H-5 Fuc), 4.86 (1 H, d, 3.5 Hz, H-l
Fuc); MS (FAB/EI) 632 (M+H)+.
(2) P~q~a~dlion of c~ ound No. B2c [Rs: CH3]
a) A solution of A1c (120 mg, 0.10 mmol), 13c (53 mg, 0.20 mmol, 2 eq)
~0 ~F
Cl , 1~ (1 3c)
F ~ F
N-hydroxysuccinimide (2.3 mg, 0.02 mmol, 0.2 eq) and abs. pyridine (31.6 mg, 0.40 mmol,
4 eq) in abs. benzene (1 ml) is heated under reflux for 2 h. The clear solution is diluted with
ethylacetate (20 ml) and extracted with HCI (0.5 n, 2 x 20 ml), NaHCO3 (20 ml) and brine
(20 ml). The solvent is removed and the residue subjected to chromatography (silicagel,
toluene/ethylacetate 5:1). Compound 14c is isolated as a colorless foam.
CO2Et
~~~~0~0
[J BzO OBn O (14c)
H3Cf~OBn OC6Hs
BnO
b) A solution of 14c (105 mg, 0.073 mmol), Bu3SnH (32 mg, 0.110 mmol, 1.5 eq), N,N'-azo-
bisisobutyronitrile (2 mg) in 3 ml of abs. benzene is heated under reflux for 1 h. Then, addi-
tional Bu3SnH (32 mg) and N,N'-azobisisobutyronitrile (2 mg) are added and heating is con-
tinued for 1 h. The solvent is removed and the residue subjected to chromatography
(toluene/ethylacetate 4:1). Compound 15c is isolated as a colorless oii.
CO2Et
-~~f~~o~c~
[J BzO OBn CH3 (15C)
~OBn
OBn
BnO
CA 022608~4 1999-01-12
W 098/06730 PCT~EP97/04279 -53-
c) A solution of 15c (15.0 mg, 0.013 mmol) in dioxane (1 ml), methanol (0.3 ml) and 2 N
NaOH (0.2 ml) is stirred at RT for 24 h. Then, 0.1 ml acetic acid is added, the solvents are
removed and the residue is passed through a short column (silicagel, isopropanollethylace-
tate/water 10:10:1). Following evaporation of the solvents the crude material is dissolved in
a mixture of methanol (1.5 ml) and acetic acid (0.1 ml). Palladium hydroxide (20%) on car-
bon (20 mg) is added and the mixture is hydrogenated at ambient pressure at RT for 16 h.
The solvents are removed and the residue is purified by reversed phase chromatography
(RP18, water/methanol 5:1~2:1). Product containing fractions are combined, the solvent is
removed, the residue is dissolved in water and passed through a sodium ion exchange
column. Following Iyophilization compound B2c is isolated as a colorless powder. 'H NMR
(400 MHz, CDCI3) ~ 0.77-1.72 (14 H, m, -CH7-cC6~", H-2ax),1.10 (3 H, d, 6.5 Hz, H-6 Puc),
1.22 (3 H, d, 6.5 Hz, H-6), 2.13 (1 H, dd (br),13.0/5.0 Hz, H-2eq), 3.19 (1 H, t, 9.0 Hz, H-4),
3.30 (1 H, dd, 9.5/3.0 Hz, H-3 Gal), 3.36 (1 H, dq, 9.0/6.5 Hz, H-5), 3.41 (1 H, t (br),12.0
Hz, H~ ), 3.51 (1 H, t, 6.0 Hz, H-5 Gal), 3.53 (1 H, dd, 9.518.0 Hz, H-2 Gal), 3.63 (2 H, d,
6.0 H~, H-6 Gal, H-6' Gal), 3.68 (1 H, dd, 10.5/4.0 Hz, H-2 Fuc), 3.72 (1 H, d (br), 3.5 Hz, H-
4 Fuc), 3.78 (1 H, dd,10.5/3.5 Hz, H-3 Fuc), 3.82 (1 H, d (br), 3.0 Hz, H-4 Gal), 3.83 - 3.97
(3 H, m, -C_-CH2-cC6H1" H-1eq, H-3), 4.41 (1 H, d, 8.0 Hz, H-1 Gal), 4.69 (1 H, q, 6.5 Hz, H-
5 Fuc), 4.95 (1 H, d, 4.0 Hz, H-1 Fuc); MS (FAB/EI) 617 (M+H)t.
(3) rle~ard~ion of compound No. B3c [R5: CO2Na]
a) To a solution of A1c (150 mg, 0.124 mmol) in abs. CH2CI2 (5 ml) 16c (158 mg,
0.373 mmol)
OAc
~r 'O
~ (16C)
o
is added and stirred at RT for 1 h. Then, an aqueous solution (20 ml) of Na2S2O3 (400 mg)
and NaHCO3 (200 mg) is added. The mixture is extracted with CH2CI2 (2 x 15 ml), the com-
bined organic layers are dried over Na2SO4 and the solvent is removed. The residue is dis-
solved in a mixture of 2-methyl-2-butene (3 ml) and tert. butanol (4 ml). An aqueous solution
(3 ml) of NaClO2 (250 mg) and NaH2PO4 (200 mg) is added and the heterogeneous mixture
is stirred vigorously for 1 h. Then, water (20 ml) and CH2C12 (20 ml) are added, the layers
CA 022608~4 1999-01-12
W O 98106730 PCT~EP97/04279
-54-
are separated and the aqueous layer is extracted with CH2C12 (2 x 20 ml). The combined
organic layers are dried with Na2SO4, the solvent is removed and the residue is subjected to
flash chromatography (silicagel, isopropanol/ethylacetate/water 25:25:5). Compound t7c is
isol~ted as a colorless foam.
CO2Et
~~~~0~0
BzO OBn CO2Na (17c)
~OBn
BnO
b) A solution of 17c (30.0 mg, 0.025 mmol) in dioxane (1.5 ml), methanol (0.5 ml) and 2 N
NaOH (0.3 ml) is stirred at RT for 24 h. Then, 0.1 ml acetic acid is added, the solvents are
removed and the residue is passed through a short column (silicagel, isopropanol/ethyl-
acetate/water 10:10:1). Following evaporation of the solvents the crude material is dissolved
in a mixture of methanol (1.5 ml) and acetic acid (0.3 ml). Palladium hydroxide (20 %) on
carbon (60 mg) is added and the mixture is hydrogenated at ambient pressure at RT for
16 h. The solvents are removed and the residue is purified by reversed phase chroi"ato-
graphy (RP18, water/methanol 5:1~2:1). Product containing fractions are combined, the
solvenl is removed, the residue is dissolved in water and passed through a sodium ion ex-
change column. Following Iyophilization compound B3c is isolated as a colorless powder.
'H NMR (400 MHz, CDCI3) ~ 0.75-1.72 (14 H, m, -CH2-cC6H11, H-1ax),1.09 (3 H, d, 6.5 Hz,
H-6 Fuc), 2.07 (1 H, m, H-2eq)1 3.27 (1 H, dd, 9.5/3.0 Hz, H-3 Gal), 3.42 (1 H, ddd,
12.0/10.0/3.0 Hz, 3.50 (1 H, dd, 9.5/8.0 Hz, H-2 Gal), 3.52 (1 H, dt, 0.5/7.5 Hz, H-5 Gal),
3.59-3.71 (6 H, m, H-4, H-5, H-2 Fuc, H-4 Fuc, H-6 Gal, H-6' Gal), 3.77 (1 H, dd,10.5/3.5
Hz, H-3 Fuc), 3.82 (1 H, dd, 3.0/0.5 Hz, H-4 Gal), 3.87 (1 H, dd,10.0t3.5 Hz, -CH-CH2-
cC6H,1), 3.93-4.02 (2 H, m, H~1~ql H-3), 4.37 (1 H, d, 8.0 Hz, H-1 Gal), 4.47 (1 H, q, 6.5 Hz,
H-5 Fuc), 4.89 (1 H, d, 4.0 Hz, H-1 Fuc); MS (FAB/EI) 617 (M+H)~.
CA 02260854 l999-0l-l2
W O 98/06730 PCT~EP97/04279
-55-
C. Biological Activities ot the Mimetics
~xa.-",le C1: Ligand Binding Assay for Det~r,~ -JtiGn of lC50 Values-cG"~e. ~ l use
of posi~ co.~ ls
This assay is performed as ~Ecclosed in Example D1 of W0 97/19,105 the contents thereof
relating to this assay being incorporated hereinwith and wherein the E-selectin/human IgG
chimera are cloned and expressed according to Kolbinger, F., Patton, J.T., Geisenhoff, G.,
Aenis, A., Li, X., Katopodis, A., Biochemistry 35:6385-6392(1996).
In this assay the compounds of formula I have an RIC50 value of from 0.01 to 1Ø
Compound No. RIC50~ Compound No. RIC
B.1a 0.013 B11b 0.019
B1b 0.017 B1c 0.024
~ RlCso means ICsO(test compound)/lC50(control compound A)
Example C2: Cell Adl.esion under Flow Co..JiliG--s
This assay is performed as disclosed in Example D3 of W0 97/19,105 the contents thereof
relating to this assay being incorporated hereinwith.
The compounds of formula I show a reduction of number of interacting cells at 50 ~M of in
the range from 40 % to 90 %.
Compound No. reduction of number Compound No. reduction of number
of interacting cells at of interacting cells at
50 ,uM 50 IlM
B.1a 61% B1c 67%
B1b 70% B11b 68%
