U.S. patent application number 15/063272 was filed with the patent office on 2016-06-30 for oligosaccharide compounds.
The applicant listed for this patent is The University of Liverpool, Victoria Link Limited. Invention is credited to Ralf Schworer, Jeremy E. Turnbull, Peter Charles Tyler, Olga Vladimirovna Zubkova.
Application Number | 20160185881 15/063272 |
Document ID | / |
Family ID | 46798422 |
Filed Date | 2016-06-30 |
United States Patent
Application |
20160185881 |
Kind Code |
A1 |
Schworer; Ralf ; et
al. |
June 30, 2016 |
Oligosaccharide Compounds
Abstract
The invention relates generally to oligosaccharide compounds and
the use of these compounds as pharmaceuticals for treating diseases
or conditions in which it is desirable to inhibit
.beta.-secretase.
Inventors: |
Schworer; Ralf; (Lower Hutt,
NZ) ; Turnbull; Jeremy E.; (Nr Cleobury Mortimer,
GB) ; Tyler; Peter Charles; (Wellington, NZ) ;
Zubkova; Olga Vladimirovna; (Kapiti, NZ) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Victoria Link Limited
The University of Liverpool |
Wellington
Liverpool |
|
NZ
GB |
|
|
Family ID: |
46798422 |
Appl. No.: |
15/063272 |
Filed: |
March 7, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14001399 |
Nov 1, 2013 |
9315587 |
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PCT/NZ2012/000035 |
Mar 9, 2012 |
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15063272 |
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61451572 |
Mar 10, 2011 |
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Current U.S.
Class: |
514/54 ;
536/53 |
Current CPC
Class: |
C08B 37/0075 20130101;
C07H 5/04 20130101; C08B 37/006 20130101; C08B 37/0078 20130101;
A61K 31/727 20130101; A61P 43/00 20180101; C08B 37/0063 20130101;
A61P 25/28 20180101; A61K 31/737 20130101; A61K 31/726
20130101 |
International
Class: |
C08B 37/00 20060101
C08B037/00; C07H 5/04 20060101 C07H005/04 |
Claims
1. An octasaccharide, decasaccharide or dodecasaccharide compound
of the formula (I): H-Q-V-W-X-Y-Z-A (I) where: A is an optionally
substituted alkoxy, aralkoxy, aryloxy group; W, X, Y and Z are each
independently a disaccharide of formula (i); V is a disaccharide of
formula (i) or V is absent; and Q is a disaccharide of formula (i)
or Q is absent ##STR00046## where: R.sup.1 is SO.sub.3H; R.sup.2 is
H; R.sup.3 is acyl; R.sup.4 is H or SO.sub.3H; and each R.sup.5 and
R.sup.6 is independently selected from COOH and H; provided that
one of R.sup.5 and R.sup.6 in each disaccharide is H and the other
is COOH; and provided that all R.sup.3 groups in the
octasaccharide, decasaccharide or dodecasaccharide are the same as
each other and provided that all R.sup.4 groups in the
octasaccharide, decasaccharide or dodecasaccharide are the same as
each other; or a pharmaceutically acceptable salt thereof.
2. A compound as claimed in claim 1 where R.sup.5 is H and R.sup.6
is COOH or a salt form of COOH.
3. A compound as claimed in claim 1 where R.sup.5 is COOH or a salt
form of COOH and R.sup.6 is H.
4. A compound as claimed in claim 1 which contains at least one
disaccharide of formula (i) where R.sup.5 is H and R.sup.6 is COOH
or a salt form of COOH and at least one disaccharide of formula (i)
where R.sup.5 is COOH or a salt form of COOH and R.sup.6 is H.
5. A compound as claimed in any one of claims 1 to 4 where the
pharmaceutically acceptable salt is an ammonium salt, a metal salt,
a salt of an organic cation, or a mixture thereof.
6. A compound as claimed in any one of claims 1 to 5 where Q and V
are absent and the compound of formula (I) is an
octasaccharide.
7. A compound as claimed in any one of claims 1 to 5 where one of Q
and V is a disaccharide of formula (i) and the other is absent and
the compound of formula (I) is a decasaccharide.
8. A compound as claimed in any one of claims 1 to 5 where Q and V
are each independently a disaccharide of formula (i) and the
compound of formula (I) is a dodecasaccharide.
9. A compound as claimed in any one of claims 1 to 8 where R.sup.3
is an acetyl group.
10. A compound as claimed in any one of claims 1 to 9 where R.sup.4
is SO.sub.3H or a salt form of SO.sub.3H.
11. A compound as claimed in any one of claims 1 to 10 where
R.sup.1 is a salt form of SO.sub.3H.
12. A compound as claimed in any one of claims 1 to 11 where A is
an optionally substituted aryloxy group.
13. A compound as claimed in claim 1, selected from the group
consisting of: ##STR00047## ##STR00048##
14. A compound as claimed in claim 1, selected from the group
consisting of ##STR00049## ##STR00050## or a pharmaceutically
acceptable salt thereof.
15. A crystalline compound of formula 13, 14, 15, 16 or 27:
##STR00051##
16. A pharmaceutical composition comprising a pharmaceutically
effective amount of a compound as claimed in any one of claims 1 to
14 and optionally a pharmaceutically acceptable carrier, diluent or
excipient.
17. A method of treating or preventing a disease or disorder in
which it is desirable to inhibit BACE-1 comprising administering a
pharmaceutically effective amount of a compound as claimed in any
one of claims 1 to 14 to a patient requiring treatment.
18. A method as claimed in claim 17 where the disease or disorder
is senile dementia, pre-senile dementia, multi-infarct dementia or
Alzheimer's disease.
Description
TECHNICAL FIELD
[0001] This invention relates generally to oligosaccharide
compounds, the use of these compounds as pharmaceuticals,
pharmaceutical compositions containing the compounds, processes for
preparing the compounds, and methods of treating diseases or
conditions in which it is desirable to inhibit
.beta.-secretase.
BACKGROUND
[0002] As populations age neurodegenerative disorders such as
Alzheimer's disease become more prevalent. Alzheimer's disease is a
common form of dementia, and is progressive and irreversible. The
pathogenesis of the disease is thought to involve cerebral deposits
of aggregated amyloid .beta.-peptide. The first (and rate-limiting)
step in the generation of amyloid .beta.-peptide is cleavage of
amyloid precursor protein by .beta.-secretase (.beta.-site amyloid
precursor protein cleaving enzyme-1, .beta.-secretase-1,
hereinafter "BACE-1"). This makes BACE-1 an attractive target for
new Alzheimer's therapies.
[0003] Heparan sulfate (HS) and its highly sulfated analogue
heparin have been shown to inhibit BACE-1 activity. HS and heparin
are both glycosaminoglycans comprising 1,4-linked disaccharide
units of .beta.-D-iduronic acid or .alpha.-L-iduronic acid with
N-acetyl-.alpha.-D-glucosamine (dominant in the case of HS) or
N-sulfo-.alpha.-D-glucosamine (dominant in the case of heparin) and
additional O-sulfate ester substituents. Heparin is a well-known
pharmaceutical with anti-coagulant activity. However, the
anti-coagulant properties of heparin need to be attenuated if it is
to be used for other pharmaceutical applications otherwise possible
side effects, such as internal bleeding and impaired blood
clotting, can be problematic.
[0004] Turnbull et al. have examined the activities of modified
heparins against BACE-1 (S. J. Patey, E. A. Edwards, E. A. Yates,
J. E. Turnbull, J. Med. Chem. 2006, 49, 6129-6132). They found
that, after porcine mucosal heparin (PIMH), the next most effective
inhibitor was a modified PIMH which had undergone N-desulfation and
N-re-acetylation. Turnbull et al. also reported the preparation of
oligosaccharides by enzymatic digestion of PIMH, and the activities
of these oligosaccharides against BACE-1 were also determined. They
found that the decasaccharide fraction was about 40-fold less
active than PIMH, and the octasaccharide fraction was 10-fold less
active than the decasaccharide fraction. Oligosaccharides
containing 18 saccharide units were about as active as PIMH itself.
As these oligosaccharide fractions are derived from a very complex
polymer, the derived fractions are not single chemical species.
[0005] WO 2007/138263 describes a study by Scholefield et al. that
showed that an N-acetylated heparin which is highly sulfated at the
2-O and 6-O positions is active against BACE-1 and that removal of
the 2-O or 6-O sulfates decreased the activity against BACE-1. WO
2007/138263 suggests that removal of one or more sulfates would
therefore be expected to have a deleterious effect on the activity.
WO 2007/138263 describes and claims such de-sulfated
oligosaccharides. WO 2007/138263 further states that neither N- nor
2-O-sulfation is an absolute requirement for high level activity
when accompanied by 6-O sulfation. As these oligosaccharide
fractions are derived from a very complex polymer, the derived
fractions are not single chemical species.
[0006] WO 2010/029185 describes oligosaccharides, including
decasaccharides and octasaccharides, which are said to be heparan
sulfate mimetics and useful in the treatment of cancer,
pathological angiogenesis and/or for inducing hematopoietic stem
cell mobilisation. These oligosaccharides have an L-iduronic acid
moiety at the non-reducing terminus.
[0007] There is a need for further oligosaccharides which are
inhibitors of BACE-1. Furthermore, if such oligosaccharides were
synthetic, in other words, if they could be synthesised de novo,
they would, advantageously, be well-characterised single chemical
entities. This would make them attractive for use as
pharmaceuticals.
[0008] It is therefore an object of the present invention to
provide oligosaccharide compounds that are inhibitors of BACE-1, or
to at least provide a useful choice.
SUMMARY OF INVENTION
[0009] In a first aspect, the present invention provides an
octasaccharide, decasaccharide or dodecasaccharide compound of the
formula (I):
H-Q-V-W-X-Y-Z-A (I)
[0010] where:
[0011] A is an optionally substituted alkoxy, aralkoxy, aryloxy
group;
[0012] W, X, Y and Z are each independently a disaccharide of
formula (i);
[0013] V is a disaccharide of formula (i) or V is absent; and
[0014] Q is a disaccharide of formula (i) or Q is absent
##STR00001##
[0015] where:
[0016] R.sup.1 is SO.sub.3H;
[0017] R.sup.2 is H;
[0018] R.sup.3 is acyl;
[0019] R.sup.4 is H or SO.sub.3H; and
[0020] each R.sup.5 and R.sup.6 is independently selected from COOH
and H; provided that one of R.sup.5 and R.sup.6 in each
disaccharide is H and the other is COOH;
[0021] and provided that all R.sup.3 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other and
provided that all R.sup.4 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other; or a
pharmaceutically acceptable salt thereof.
[0022] Preferably R.sup.5 is H and R.sup.6 is COOH. Alternatively
it is preferred that R.sup.5 is COOH and R.sup.6 is H.
Alternatively, the octasaccharide, decasaccharide or
dodecasaccharide contains at least one disaccharide of formula (i)
where R.sup.5 is H and R.sup.6 is COOH and at least one
disaccharide of formula (i) where R.sup.5 is COOH and R.sup.6 is
H.
[0023] In a second aspect, the present invention provides an
octasaccharide, decasaccharide or dodecasaccharide compound of the
formula (Ia):
H-Q-V-W-X-Y-Z-A (Ia)
[0024] where:
[0025] A is an optionally substituted alkoxy, aralkoxy, aryloxy
group;
[0026] W, X, Y and Z are each independently a disaccharide of
formula (i);
[0027] V is a disaccharide of formula (i) or V is absent; and
[0028] Q is a disaccharide of formula (i) or Q is absent
##STR00002##
[0029] where:
[0030] R.sup.1 is SO.sub.3H;
[0031] R.sup.2 is H;
[0032] R.sup.3 is acyl;
[0033] R.sup.4 is H or SO.sub.3H;
[0034] R.sup.5 is H; and
[0035] R.sup.6 is COOH;
[0036] provided that all R.sup.3 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other and
provided that all R.sup.4 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other; or a
pharmaceutically acceptable salt thereof.
[0037] In a third aspect, the present invention provides an
octasaccharide, decasaccharide or dodecasaccharide compound of the
formula (Ib):
H-Q-V-W-X-Y-Z-A (Ib)
[0038] where:
[0039] A is an optionally substituted alkoxy, aralkoxy, aryloxy
group;
[0040] W, X, Y and Z are each independently a disaccharide of
formula (i);
[0041] V is a disaccharide of formula (i) or V is absent; and
[0042] Q is a disaccharide of formula (i) or Q is absent
##STR00003##
[0043] where:
[0044] R.sup.1 is SO.sub.3H;
[0045] R.sup.2 is H;
[0046] R.sup.3 is acyl;
[0047] R.sup.4 is H or SO.sub.3H;
[0048] R.sup.5 is COOH; and
[0049] R.sup.6 is H;
[0050] provided that all R.sup.3 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other and
provided that all R.sup.4 groups in the octasaccharide,
decasaccharide or dodecasaccharide are the same as each other; or a
pharmaceutically acceptable salt thereof.
[0051] Preferably the pharmaceutically acceptable salt is an
ammonium salt, a metal salt, e.g. a sodium salt, or a salt of an
organic cation, or a mixture thereof.
[0052] In some examples, Q and V are absent and the compound of
formula (I), (Ia) or (Ib) is an octasaccharide.
[0053] In other examples, one of Q or V is absent and the other is
a disaccharide of formula (i) and the compound of formula (I), (Ia)
or (Ib) is a decasaccharide.
[0054] In still other examples, Q and V are each independently a
disaccharide of formula (i) and the compound of formula (I), (Ia)
or (Ib) is a dodecasaccharide.
[0055] Preferably R.sup.3 is a lower acyl group, e.g. an acetyl
group.
[0056] Preferably R.sup.4 is SO.sub.3H or a salt form thereof, e.g.
R.sup.4 may be SO.sub.3Na or SO.sub.3NH.sub.4. Alternatively it is
preferred that R.sup.4 is H.
[0057] In some examples R.sup.1 is a salt form of SO.sub.3H, e.g.
R.sup.1 may be SO.sub.3NH.sub.4 or SO.sub.3Na.
[0058] In some examples R.sup.5 or R.sup.6 is a salt form of COOH,
e.g. R.sup.5 or R.sup.6 may be COONa or COONH.sub.4.
[0059] Preferably A is an optionally substituted aryloxy group,
such as an aryloxy group, e.g. a phenoxy group, substituted with an
alkoxy group, e.g. a lower alkoxy group, e.g. a 4-methoxyphenoxy
group.
[0060] Alternatively A is an optionally substituted alkoxy group,
preferably hexyloxy or octyloxy group, or an
.omega.-(N-benzyloxycarbonylamino)-alkyloxy group, preferably an
8-(N-benzyloxycarbonylamino)-octyl, or
6-(N-benzyloxycarbonylamino)hexyl group, or an
.omega.-aminoalkyloxy group, preferably an 8-aminooctyl or
6-aminohexyl group, or a methoxy-per(ethyleneoxy) group, preferably
a methoxy-tris(ethylenoxy)-group.
[0061] In another aspect the invention provides a compound selected
from the group consisting of:
##STR00004## ##STR00005##
[0062] or a pharmaceutically acceptable salt thereof.
[0063] In another aspect the invention provides a compound selected
from the group consisting of:
##STR00006## ##STR00007##
[0064] or a pharmaceutically acceptable salt thereof.
[0065] The invention also provides an acid form of any one of the
compounds (a) to (g) or (j) to (p) above.
[0066] In another aspect, the invention provides a prodrug, e.g. an
ester prodrug, of a compound of formula (I), (Ia) or (Ib).
[0067] In another aspect the invention provides a compound of
formula 13, 14, 15, 16 or 27:
##STR00008##
[0068] In another aspect the invention provides a crystalline
compound of formula 13, 14, 15, 16 or 27:
##STR00009##
[0069] In another aspect, the invention provides a crystalline
compound of formula 13, having a melting point of about 131.degree.
C.
##STR00010##
[0070] In another aspect, the invention provides a crystalline
compound of formula 14, having a melting point of about
140-141.degree. C.
##STR00011##
[0071] In another aspect, the invention provides a crystalline
compound of formula 16, having a melting point of about 144.degree.
C.
##STR00012##
[0072] In another aspect, the invention provides a crystalline
compound of formula 27, having a melting point of about 144.degree.
C.
##STR00013##
[0073] In another aspect, the invention provides a crystalline
compound of formula 13, having: [0074] i. a crystal structure as
shown in FIG. 1; and/or [0075] ii. crystal lattice parameters at
123(2) K of: a=10.454(2) .ANG., b=35.610(7) .ANG., c=14.408(3)
.ANG., .alpha.=90.degree., .beta.=95.61(3).degree.,
.gamma.=90.degree.; and/or [0076] iii. a crystal structure
belonging to a monoclinic space group, e.g. P2.sub.1
[0077] determined by X-ray crystal structure analysis
##STR00014##
[0078] In another aspect, the invention provides a crystalline
compound of formula 14, having: [0079] i. a crystal structure as
shown in FIG. 2; and/or [0080] ii. crystal lattice parameters at
164(2) K of: a=9.2713(7) .ANG., b=17.4067(11) .ANG., c=15.0036(11)
.ANG., a=90.degree., .beta.=97.449(7).degree., .gamma.=90.degree.;
and/or [0081] iii. a crystal structure belonging to a monoclinic
space group, e.g. P2.sub.1
[0082] determined by X-ray crystal structure analysis
##STR00015##
[0083] In another aspect, the invention provides a crystalline
compound of formula 15, having: [0084] i. a crystal structure as
shown in FIG. 3; and/or [0085] ii. crystal lattice parameters at
118(2) K of: a=38.3346(13) .ANG., b=8.0744(3) .ANG., c=16.1659(6)
.ANG., .alpha.=90.degree., .beta.=91.222(2).degree.,
.gamma.=90.degree.; and/or [0086] iii. a crystal structure
belonging to a monoclinic space group, e.g. C2
[0087] determined by X-ray crystal structure analysis
##STR00016##
[0088] In another aspect, the invention provides a crystalline
compound of formula 16, having: [0089] i. a crystal structure as
shown in FIG. 4; and/or [0090] ii. crystal lattice parameters at
123(2) K of: a=14.8343(11) .ANG., b=8.4771(6) .ANG., c=21.8112(17)
.ANG., .alpha.=90.degree., .beta.=91.780(7).degree.,
.gamma.=90.degree.; and/or [0091] iii. a crystal structure
belonging to a monoclinic space group, e.g. P2.sub.1
[0092] determined by X-ray crystal structure analysis
##STR00017##
[0093] In another aspect, the invention provides a crystalline
compound of formula 27, having: [0094] i. a crystal structure as
shown in FIG. 5; and/or [0095] ii. crystal lattice parameters at
164(2) K of: a=8.1104(2) .ANG., b=19.5548(6) .ANG., c=27.2321(19)
.ANG., .alpha.=90.degree., .beta.=90.degree., .gamma.=90.degree.;
and/or [0096] iii. a crystal structure belonging to a orthorhombic
space group, e.g. P2.sub.12.sub.12.sub.1
[0097] determined by X-ray crystal structure analysis
##STR00018##
[0098] In another aspect the invention provides the use of a
compound of formula 13, 14, 15, 16 or 27 for preparing a compound
of formula (I), (Ia) or (Ib).
[0099] In another aspect the invention provides a composition
comprising a pharmaceutically effective amount of a compound of
formula (I), (Ia) or (Ib) and optionally a carrier.
[0100] In another aspect the invention provides a pharmaceutical
composition comprising a pharmaceutically effective amount of a
compound of formula (I), (Ia) or (Ib) and optionally a
pharmaceutically acceptable carrier, diluent or excipient.
[0101] In another aspect the invention provides a compound of
formula (I), (Ia) or (Ib) in combination with at least one other
compound, e.g. a second drug compound. The other compound may be,
for example, an oligosaccharide compound, a cyclitol such as
scyllo-inositol or D-chiro-inositol, an acetylcholinesterase
inhibitor, a nicotinic agonist, an antibody targeting
.beta.-amyloid, an inhibitor of .beta.-amyloid, an inhibitor of tau
aggregation or memantine.
[0102] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) for inhibiting BACE-1.
[0103] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) as a medicament.
[0104] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) for treating or preventing a
disease or disorder in which it is desirable to inhibit BACE-1.
[0105] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) for treating or preventing a
neurodegenerative disorder such as senile dementia, pre-senile
dementia, multi-infarct dementia or Alzheimer's disease, preferably
Alzheimer's disease.
[0106] In another aspect the invention provides the use of a
pharmaceutical composition comprising a pharmaceutically effective
amount of a compound of formula (I), (Ia) or (Ib) for treating or
preventing a disease or disorder in which it is desirable to
inhibit BACE-1.
[0107] In another aspect the invention provides the use of a
pharmaceutical composition comprising a pharmaceutically effective
amount of a compound of formula (I), (Ia) or (Ib) for treating or
preventing a neurodegenerative disorder such as senile dementia,
pre-senile dementia, multi-infarct dementia or Alzheimer's disease,
preferably Alzheimer's disease.
[0108] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) for use in the manufacture of
a medicament.
[0109] In another aspect the invention provides a pharmaceutical
composition for treating or preventing a disease or disorder in
which it is desirable to inhibit BACE-1 comprising a compound of
formula (I), (Ia) or (Ib).
[0110] In another aspect the invention provides a pharmaceutical
composition for treating or preventing a neurodegenerative disorder
such as senile dementia, pre-senile dementia, multi-infarct
dementia or Alzheimer's disease, preferably Alzheimer's disease,
comprising a compound of formula (I), (Ia) or (Ib).
[0111] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) in the manufacture of a
medicament for the treatment or prevention of a disease or disorder
in which it is desirable to inhibit BACE-1.
[0112] In another aspect the invention provides a method of
treating or preventing a disease or disorder in which it is
desirable to inhibit BACE-1 comprising administering a
pharmaceutically effective amount of a compound of formula (I),
(Ia) or (Ib) to a patient requiring treatment.
[0113] In another aspect the invention provides a method of
treating or preventing a neurodegenerative disorder such as senile
dementia, pre-senile dementia, multi-infarct dementia or
Alzheimer's disease, preferably Alzheimer's disease, comprising
administering a pharmaceutically effective amount of a compound of
formula (I), (Ia) or (Ib) to a patient requiring treatment.
[0114] In another aspect the invention provides the use of a
compound of formula (I), (Ia) or (Ib) in combination with at least
one other compound, e.g. a second drug compound, e.g. an
oligosaccharide compound, a cyclitol such as scyllo-inositol or
D-chiro-inositol, an acetylcholinesterase inhibitor, a nicotinic
agonist, an antibody targeting .beta.-amyloid, an inhibitor of
.beta.-amyloid, an inhibitor of tau aggregation or memantine, for
treating or preventing a disease or disorder in which it is
desirable to inhibit BACE-1 (e.g. a neurodegenerative disorder such
as senile dementia, pre-senile dementia, multi-infarct dementia or
Alzheimer's disease, preferably Alzheimer's disease).
[0115] In another aspect the invention provides a method of
treating or preventing a disease or disorder in which it is
desirable to inhibit BACE-1 (e.g. a neurodegenerative disorder such
as senile dementia, pre-senile dementia, multi-infarct dementia or
Alzheimer's disease, preferably Alzheimer's disease) comprising
administering a pharmaceutically effective amount of a compound of
formula (I), (Ia) or (Ib) in combination with at least one other
compound, e.g. a second drug compound, e.g. an oligosaccharide
compound, a cyclitol such as scyllo-inositol or D-chiro-inositol,
an acetylcholinesterase inhibitor, a nicotinic agonist, an antibody
targeting .beta.-amyloid, an inhibitor of .beta.-amyloid, an
inhibitor of tau aggregation or memantine. The compound of formula
(I), (Ia) or (Ib) and the other compound may be administered
separately, simultaneously or sequentially.
[0116] The diseases or disorders include neurodegenerative
disorders such as senile dementia, pre-senile dementia,
multi-infarct dementia or Alzheimer's disease, preferably
Alzheimer's disease. 15
[0117] The compound of formula (I), (Ia) or (Ib) may be selected
from the group consisting of compounds (a) to (g) and (j) to (p) as
defined above.
[0118] Compounds of formulae (I), (Ia) or (Ib) are hereinafter
described as "compounds of the invention". A compound of the
invention includes a compound in any form, e.g. in free form or in
the form of a salt or a solvate. For example, the compounds of the
invention, e.g. the compounds of formula (I), (Ia), (Ib) and the
compounds (a) to (g) and (j) to (p) can exist as the free acid form
and the invention is intended to cover such acid forms.
[0119] It will be appreciated that any of the sub-scopes disclosed
herein, e.g. with respect to R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, A, Q, V, W, X, Y and Z may be combined with any
of the other sub-scopes disclosed herein to produce further
sub-scopes.
BRIEF DESCRIPTION OF THE FIGURES
[0120] FIG. 1 shows an ORTEP diagram of compound 13.
[0121] FIG. 2 shows an ORTEP diagram of compound 14.
[0122] FIG. 3 shows an ORTEP diagram of compound 15.
[0123] FIG. 4 shows an ORTEP diagram of compound 16.
[0124] FIG. 5 shows an ORTEP diagram of compound 27.
DETAILED DESCRIPTION
[0125] Definitions
[0126] The term "alkyl" means any saturated hydrocarbon radical
having up to 30 carbon atoms and includes any C.sub.1-C.sub.25,
C.sub.1-C.sub.20, C.sub.1-C.sub.15, C.sub.1-C.sub.10, or
C.sub.1-C.sub.6 alkyl group, and is intended to include cyclic,
straight- and branched-chain alkyl groups. Cyclic alkyl groups
include those groups having one or more ring oxygen atoms. Examples
of alkyl groups include: methyl group, ethyl group, n-propyl group,
iso-propyl group, n-butyl group, iso-butyl group, sec-butyl group,
t-butyl group, n-pentyl group, 1,1-dimethylpropyl group,
1,2-dimethylpropyl group, 2,2-dimethylpropyl group, 1-ethylpropyl
group, 2-ethylpropyl group, n-hexyl group, 1,2-dimethylbutyl group,
cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl
group, cycloheptyl group, cyclooctyl group, tetrahydrofuranyl group
and tetrahydropyranyl group.
[0127] The term "lower alkyl" means any saturated hydrocarbon
radical having from 1 to 6 carbon atoms and is intended to include
cyclic, straight- and branched-chain alkyl groups.
[0128] The term "alkoxy" means --OR where R is alkyl as defined
above. The term "lower alkoxy" means --OR where R is lower alkyl as
defined above.
[0129] Any alkoxy group may optionally be substituted with one or
more substituents selected from the group consisting of fluorine,
chlorine, methoxy, ethoxy,
[0130] O(CH.sub.2CH.sub.2).sub.nOMe where n is 1-10, NH.sub.2,
NHCO.sub.2Bn, and CO.sub.2H or pharmaceutically acceptable salt
form thereof.
[0131] The term "aryl" means an aromatic radical having 4 to 18
carbon atoms and includes heteroaromatic radicals. Examples include
monocyclic groups, as well as fused groups such as bicyclic groups
and tricyclic groups. Some examples include phenyl group, indenyl
group, 1-naphthyl group, 2-naphthyl group, azulenyl group,
heptalenyl group, biphenyl group, indacenyl group, acenaphthyl
group, fluorenyl group, phenalenyl group, phenanthrenyl group,
anthracenyl group, cyclopentacyclooctenyl group, and
benzocyclooctenyl group, pyridyl group, pyrrolyl group, pyridazinyl
group, pyrimidinyl group, pyrazinyl group, triazolyl group
(including a 1-H-1,2,3-triazol-1-yl and a 1-H-1,2,3-triazol-4-yl
group), tetrazolyl group, benzotriazolyl group, pyrazolyl group,
imidazolyl group, benzimidazolyl group, indolyl group, isoindolyl
group, indolizinyl group, purinyl group, indazolyl group, furyl
group, pyranyl group, benzofuryl group, isobenzofuryl group,
thienyl group, thiazolyl group, isothiazolyl group, benzothiazolyl
group, oxazolyl group, and isoxazolyl group.
[0132] Any aryl group may optionally be substituted with one or
more substituents selected from the group consisting of fluorine,
chlorine, alkoxy (including methoxy and ethyoxy), alkyl (including
methyl and ethyl), cyano, acylamino, azido or NHCO.sub.2Bn.
[0133] The term "aryloxy" means --OR' where R' is aryl as defined
above.
[0134] The term "aralkyl" means an aryl group covalently linked to
an alkylene group.
[0135] The term "aralkoxy" means --OR'' where R'' is aralkyl as
defined above.
[0136] Any aralkoxy group may optionally be substituted with one or
more substituents selected from the group consisting of fluorine,
chlorine, alkoxy (including methoxy and ethyoxy), alkyl (including
methyl and ethyl), cyano, acylamino, azido or NHCO.sub.2Bn.
[0137] The term "acyl" means
##STR00019##
where R''' is alkyl, aralkyl or aryl as defined above. The term
"lower acyl" means a C.sub.2-C.sub.6 acyl group, having a
corresponding meaning to "lower alkyl" as defined above.
[0138] The term "halogen" includes fluorine, chlorine, bromine and
iodine.
[0139] The term "prodrug" as used herein means a pharmacologically
acceptable derivative of the compounds of formulae (I), (Ia) and
(Ib), such that an in vivo biotransformation of the derivative
gives the compound as defined in formulae (I), (Ia) and (Ib).
Prodrugs of compounds of formulae (I), (Ia) and (Ib) may be
prepared by modifying functional groups present in the compounds in
such a way that the modifications are cleaved in vivo to give the
parent compound. Typically, prodrugs of the compounds of formulae
(I), (Ia) and (Ib) will be ester prodrug forms.
[0140] The term "pharmaceutically acceptable salts" is intended to
apply to non-toxic salts such as ammonium salts, metal salts, e.g.
sodium salts, or salts of organic cations, or a mixture
thereof.
[0141] The term "protecting group" means a group that selectively
protects an organic functional group, temporarily masking the
chemistry of that functional group and allowing other sites in the
molecule to be manipulated without affecting the functional group.
Suitable protecting groups are known to those skilled in the art
and are described, for example, in Protective Groups in Organic
Synthesis (3.sup.rd Ed.), T. W. Greene and P. G. M. Wuts, John
Wiley & Sons Inc (1999). Examples of protecting groups include,
but are not limited to: O-benzyl, O-benzhydryl, O-trityl,
O-tert-butyldimethylsilyl, O-tert-butyldiphenylsilyl,
O-4-methylbenzyl, O-acetyl, O-chloroacetyl, O-methoxyacetyl,
O-benzoyl, O-4-bromobenzoyl, O-4-methylbenzoyl,
O-fluorenylmethoxycarbonyl and O-levulinoyl.
[0142] The term "patient" includes human and non-human animals.
[0143] The terms "treatment", "treating" and the like include the
alleviation of one or more symptoms, or improvement of a state
associated with the disease or disorder, for example, improvement
in cognition, improvement in memory function.
[0144] The terms "preventing", "prevention" and the like include
the prevention of one or more symptoms associated with the disease
or disorder.
[0145] The Compounds of the Invention
[0146] The compounds of the invention, particularly those
exemplified, are inhibitors of BACE-1 and are useful as
pharmaceuticals, particularly for the treatment or prevention of
diseases or conditions in which it is desirable to inhibit BACE-1,
e.g. neurodegenerative disorders such as senile dementia,
pre-senile dementia, multi-infarct dementia or Alzheimer's disease,
particularly Alzheimer's disease. The compounds of the invention
are useful in both free base form and in the form of salts and/or
solvates.
[0147] Those skilled in the art will appreciate that the compounds
of the invention can exist as stereoisomers. For example, depending
on the stereochemistry at the carbon marked (#) in the disaccharide
of formula (i), the disaccharide can be either gluco or ido form.
Thus, each R.sup.5 and each R.sup.6 in each disaccharide of formula
(i) is independently selected from COOH and H; provided that one of
R.sup.5 and R.sup.6 in each disaccharide is H and the other is
COOH.
##STR00020##
[0148] The octasaccharides, decasaccharides and dodecasaccharides
of the invention are made up of the disaccharides of formula (i).
The skilled person will therefore further appreciate that the
octasaccharides, decasaccharides and dodecasaccharides of the
invention may be all gluco form (where all disaccharides of formula
(i) in the compound are gluco), all ido form (where all
disaccharides of formula (i) in the compound are ido) or a mixture
of gluco and ido forms (where the disaccharides of formula (i) in
the compound are a mixture of gluco and ido).
[0149] As described in Example 4, heparin has an IC.sub.50 of 0.002
.mu.g/mL against human recombinant BACE-1 and N-acetylated low
molecular weight heparin (NAcLMWH) has an IC.sub.50 of 0.007
.mu.g/mL. Surprisingly, the oligosaccharides of the invention are
potent inhibitors of BACE-1. For example, compounds 90, 91 and 92
all have an IC.sub.50 of about 0.01 .mu.g/mL against human
recombinant BACE-1. Indeed, some compounds of the invention are
only approximately 5-fold less potent than heparin by mass. This is
surprising when compared to the activities of the octasaccharide
and decasaccharide fractions reported in J. Med. Chem. 2006, 49,
6129-6132, and indicates a role for synthetic oligosaccharides in
treating diseases or disorders where it is desirable to inhibit
BACE-1. The synthetic oligosaccharides of the invention also have
the advantage that they are discrete chemical entities of known
structure.
[0150] Interestingly, not only are the 6-sulfated compounds of the
invention surprisingly potent synthetic oligosaccharides, but, even
more surprisingly, the synthetic oligosaccharides of the invention
which are sulfated at the 2- and 6-positions tend to be
significantly more active than the corresponding 6-sulfated
oligosaccharides. This would not have been predicted based on
previous studies on modified full length heparins.
[0151] Advantageously, the compounds of the invention have
attentuated anti-coagulent activity. Referring to Example 5, none
of the compounds of the invention display any measurable ability to
accelerate antithrombin-III mediated inactivation of Factor Xa, as
measured by cleavage of a peptide substrate.
[0152] The compounds of the invention may be administered to a
patient by a variety of routes, including orally, parenterally, by
inhalation spray, topically, rectally, nasally, buccally or via an
implanted reservoir. The compounds may also be administered by
intracerebral, intracerebroventricular or intrathecal delivery. For
parenteral administration, injections may be given intravenously,
intra-arterially, intramuscularly or subcutaneously.
[0153] The amount of a compound of the invention to be administered
to a patient will vary widely according to the nature of the
patient and the nature and extent of the disorder to be treated.
Typically the dosage for an adult human will be in the range of
about 0.01 .mu.g/kg to about 1 g/kg, preferably about 0.01 mg/kg to
about 100 mg/kg. The specific dosage required for any particular
patient will depend upon a variety of factors, such as the
patient's age, body weight, general health, gender and diet.
Optimal doses will depend on other factors such as mode of
administration and level of progression of the disease or disorder.
Doses may be given once daily, or two or more doses may be required
per day. For example, a dosage regime for an Alzheimer's patient
might require one dose in the morning and one in the evening.
Alternatively, a dosage regime for such a patient might require
four hourly doses.
[0154] For oral administration the compounds can be formulated into
solid or liquid preparations, for example tablets, capsules,
granules, powders, solutions, suspensions, syrups, elixirs and
dispersions. Such preparations are well known in the art as are
other oral dosage regimes not listed here.
[0155] For parenteral administration, compounds of the invention
can be formulated into sterile solutions, emulsions and
suspension.
[0156] Compounds of the invention may be mixed with suitable
vehicle and then compressed into the desired shape and size. The
compounds may be tableted with conventional tablet bases such as
lactose, sucrose and corn starch, together with a binder, a
disintegration agent and a lubricant. The binder may be, for
example, corn starch or gelatin, the disintegrating agent may be
potato starch or alginic acid, and the lubricant may be magnesium
stearate. For oral administration in the form of capsules, diluents
such as lactose and dried cornstarch may be employed. Other
components such as colourings, sweeteners or flavourings may be
added. Tablets, capsules or powders for oral administration may
contain up to about 99% of a compound of the invention.
[0157] When liquid preparations are required for oral use, a
compound of the invention may be combined with a pharmaceutically
acceptable carriers such as water, an organic solvent such as
ethanol, or a mixture of both, and optionally other additives such
as emulsifying agents, suspending agents, buffers, preservatives,
and/or surfactants may be used. Colourings, sweeteners or
flavourings may also be added.
[0158] The compounds may also be administered by injection in a
pharmaceutically acceptable diluent such as water or saline. The
diluent may comprise one or more other ingredients such as ethanol,
propylene glycol, an oil or a pharmaceutically acceptable
surfactant.
[0159] The compounds of the invention may also be administered
topically. Carriers for topical administration of the compounds
include mineral oil, liquid petrolatum, white petrolatum, propylene
glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax
and water. The compounds may be present as ingredients in lotions
or creams, for topical administration to skin or mucous membranes.
Such creams may contain the active compounds suspended or dissolved
in one or more pharmaceutically acceptable carriers. Suitable
carriers include mineral oil, sorbitan monostearate, polysorbate
60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl
alcohol and water.
[0160] The compounds of the invention may further be administered
by means of sustained release systems. For example, they may be
incorporated into a slowly dissolving tablet or capsule.
[0161] Synthesis of the Compounds of the Invention
[0162] The compounds of the invention may be prepared by a variety
of different methods. The following are representative non-limiting
general methods for synthesising compounds of the invention.
[0163] The synthetic strategy involves disaccharide building blocks
as intermediates for the preparation of the compounds of the
invention. The present invention therefore also relates to
intermediates and methods for the synthesis of compounds of the
invention.
[0164] The Disaccharide Building Blocks
[0165] The octa- deca- and dodecasaccharide compounds of the
invention are prepared from three, four or five neutral
disaccharide building blocks of general formula (A) and/or (B),
respectively, selected independently for the reducing terminal and
internal disaccharide units, and one of (C) or (D) selected
independently for the non-reducing terminal disaccharide unit.
##STR00021##
[0166] wherein R.sup.1', R.sup.2', R.sup.3', R.sup.4' and R.sup.7'
are protecting groups chosen with differing reactivity so that they
can be selectively removed as required in the process detailed
below, namely R.sup.4' before R.sup.7' before R.sup.1' before
R.sup.2' before R.sup.3'.
[0167] Ester groups are suitable R.sup.1', R.sup.2' and R.sup.7'
protecting groups. The ester protecting group R.sup.2' facilitates
the control of the anomeric stereochemistry of the glycosylation
reactions to give products with the required
1,2-trans-stereochemistry with high selectivity.
[0168] The acetyl group is a suitable R.sup.1' protecting
group.
[0169] The benzoyl group is a suitable R.sup.2' protecting
group.
[0170] The benzyl group is a suitable R.sup.3' protecting
group.
[0171] The chloroacetyl group is a suitable R.sup.7' protecting
group. The methoxyacetyl and levulinoyl groups are alternative
R.sup.7' protecting groups. Levulinoyl groups are described by Wang
et al., Chem. J. Eur., 16 (2010) 8365.
[0172] The fluorenylmethoxycarbonyl group is a suitable R.sup.4'
protecting group.
[0173] Advantageously and surprisingly, some disaccharide building
blocks are crystalline compounds. This makes them particularly
suitable intermediates for the synthesis of compounds of the
invention, as they can be easily stored and transported. The
present invention also relates to such crystalline disaccharide
intermediates.
[0174] Synthesis of a 4-Methoxyphenyl Glycoside Octasaccharide
Compound of the Invention
[0175] An octasaccharide compound of the invention is synthesised
from three neutral disaccharide building blocks of general formula
(A) and/or (B), respectively, selected independently for the
reducing terminal and internal disaccharide units, and one of (C)
or (D) selected independently for the non-reducing terminal
disaccharide unit.
[0176] Stage 1, the First Glycosylation Reaction
[0177] The tetrasaccharide (G) is prepared by: [0178] a) synthesis
of an acceptor (E) from a building block (A) or (B) by selective
removal of the protecting group R.sup.4'; [0179] b) synthesis of a
glycosyl donor (F) from a building block (A) or (B) by selective
removal of the 4-methoxyphenyl group and introducing a suitable
leaving group at C-1 of the reducing sugar; and [0180] c) coupling
of the donor (F) and acceptor (E).
##STR00022##
[0181] where R.sup.8' and R.sup.9' are independently chosen from
hydrogen and CH.sub.2OR.sup.7' with the proviso that when one is
hydrogen, the other is CH.sub.2OR.sup.7'.
[0182] Selective removal of a fluorenylmethoxycarbonyl protecting
group is conveniently achieved by dissolution in mixture of
dichloromethane and triethylamine, typically about 4:1 v/v, 20 mL
per mmol, at ambient temperature, typically in the range of about
10.degree. C. to about 30.degree. C.
[0183] Selective removal of the 4-methoxyphenyl group is
conveniently achieved by use of CAN.
[0184] The trichloroacetimidate group is a suitable leaving group
X. The methylthio, phenylthio and 4-methylphenylthio groups are
alternative leaving groups X.
[0185] Glycosyl trichloroacetimidates are conveniently synthesised
from the free-sugar precursors by reaction with excess
trichloroacetonitrile and a base. NaH is a suitable base. DBU is
alternative base (see WO 03/022860). DCM is a suitable solvent and
the reaction can be conducted in the range of about -10.degree. C.
to about 40.degree. C., preferably about 0.degree. C. to about
10.degree. C., typically in an ice bath.
[0186] Suitable glycosidation conditions are described below.
[0187] Syntheses of suitable 1-thioglycoside donors and conditions
suitable for glycosylation using them are reported in Wang et al.,
Chem. J. Eur., 16 (2010) 8365 and in WO 03/0228860.
[0188] Stage 2, the Second Glycosylation Reaction
[0189] The hexasaccharide (H) is prepared by: [0190] a) synthesis
of a tetrasaccharide acceptor by selective removal of the
protecting group R.sup.4' from (G) to generate a free secondary
hydroxy group; and [0191] b) coupling of the donor (F) from Stage 1
to this tetrasaccharide acceptor.
##STR00023##
[0192] where R.sup.10' and R.sup.11' are independently chosen from
hydrogen and CH.sub.2OR.sup.7' with the proviso that when one is
hydrogen, the other is CH.sub.2OR.sup.7'.
[0193] Stage 3, the Third Glycosylation Reaction
[0194] The octasaccharide (J) is prepared by: [0195] a) synthesis
of a hexasaccharide acceptor by selective removal of the protecting
group R.sup.4' from (H) to generate a free secondary hydroxy group;
[0196] b) synthesis of a glycosyl donor (I) from a building block
(C) or (D) by selective removal of the 4-methoxyphenyl group and
introducing a suitable leaving group at C-1 of the reducing sugar;
and [0197] c) coupling of the donor (I) with this hexasaccharide
acceptor.
##STR00024##
[0198] where R.sup.12' and R.sup.13' are independently chosen from
hydrogen and CH.sub.2OR.sup.7' with the proviso that when one is
hydrogen, the other is CH.sub.2OR.sup.7'.
[0199] Stage 4, Conversion to Final Product
[0200] The octasaccharide of the invention is prepared by: [0201]
a) selective removal of all protecting groups R.sup.7' from the
neutral octasaccharide derivative (J); [0202] b) oxidation of all
the resulting free primary hydroxy groups to the corresponding
carboxylic acid groups (or salt forms thereof); [0203] c)
conversion of all the carboxylic acid moieties into their
corresponding methyl esters; [0204] d) converting all the
azide-groups into the corresponding acylamino-groups; [0205] e)
selective removal of all R.sup.1' protecting groups; [0206] f)
sulfation of the resulting primary hydroxy groups; [0207] g)
selective removal of all R.sup.2' protecting groups and hydrolysis
of the methyl esters; [0208] h) sulfation of the resulting
secondary hydroxy groups, or proceeding directly to step i); [0209]
i) removal of the R.sup.3' protecting groups; and [0210] j)
conversion to the desired cationic salt form of the final
product.
[0211] Selective removal of the chloroacetyl R.sup.7' protecting
group is conveniently achieved using DABCO (6 equiv. per
chloroacetyl group) in dry ethanol heated at about 60.degree. C. to
about 70.degree. C. under argon.
[0212] Oxidation of the resulting free primary hydroxy group is
conveniently achieved using the TEMPO-BAIB system in aqueous
acetonitrile at room temperature. Conversion of the resulting
carboxylic acid into corresponding methyl ester is conveniently
achieved by reaction with diazomethane, TMS-diazomethane, or a
combination of iodomethane and a base, conveniently potassium
bicarbonate, in DMF. An alternative TEMPO oxidation system is
described in WO 03/022860.
[0213] Conversion of the azido-group into the corresponding
acetamido-group is conveniently achieved by reaction with
thiolacetic acid in dry pyridine at room temperature. Conversion of
the azido-group into an alternative acylamido-group is achieved by
reduction of the azido-group, conveniently with trialkylphosphine
(e.g. nBu.sub.3P), triarylphosphine (e.g. Ph.sub.3P), or a metal
catalyst (e.g. Pd/C) and reaction, either in the same reaction
mixture or separately, with an acylating agent (e.g. an acyl
anhydride or an acyl chloride).
[0214] Selective removal of the acetyl R.sup.1' protecting group is
conveniently achieved using HCl in a DCM-MeOH solution in the
temperature range of about 0.degree. C. to about 30.degree. C. The
HCl can be generated in situ by use of acetyl chloride.
[0215] Cleavage of ester R.sup.2' protecting groups is conveniently
conducted by saponification with sodium hydroxide in aqueous
methanol.
[0216] Sulfation reactions are conveniently conducted using sulfur
trioxide trimethylamine complex (5 equiv per hydroxyl group) in dry
DMF at about 50.degree. C. to about 60.degree. C. under argon.
[0217] Removal of the benzyl R.sup.3' protecting group is
conveniently conducted using hydrogen and a palladium catalyst,
typically palladium hydroxide on carbon, in aqueous tetrahydrofuran
in the range of about 10.degree. C. to about 30.degree. C.
[0218] Final products are converted into the desired salt form by
elution with water through a Strong acid ion exchange resin column,
e.g. Dowex 50WX8-200 (Dow Chemical Company, USA) in the desired
salt form, e.g. Na.sup.+.
[0219] Synthesis of a 4-Methoxyphenyl Glycoside Decasaccharide of
the Invention
[0220] A decasaccharide compound of the invention is synthesised
from four neutral disaccharide building blocks of general formula
(A) and/or (B), respectively, selected independently for the
reducing terminal and internal disaccharide units, and one of (C)
or (D) selected independently for the non-reducing terminal
disaccharide unit.
[0221] Thus, the octasaccharide (K) is prepared by: [0222] a)
synthesis of a hexasaccharide acceptor by selective removal of the
protecting group R.sup.4' from hexasaccharide derivative (H) to
generate a free secondary hydroxy group; [0223] b) coupling of the
donor (F) with this hexasaccharide acceptor.
##STR00025##
[0224] Then decasaccharide (L) is prepared by: [0225] a) synthesis
of an octasaccharide acceptor by selective removal of the
protecting group R.sup.4' from octasaccharide derivative (K) to
generate a free secondary hydroxy group; [0226] b) coupling of the
donor (I) with this octasaccharide acceptor.
##STR00026##
[0227] where R.sup.14' and R.sup.15' are independently chosen from
hydrogen and CH.sub.2OR.sup.7' with the proviso that when one is
hydrogen, the other is CH.sub.2OR.sup.7'.
[0228] Finally, the decasaccharide of the invention is prepared by:
[0229] a) selective removal of all protecting groups R.sup.7' from
the neutral decasaccharide derivative (L); [0230] b) oxidation of
all the resulting free primary hydroxy groups to the corresponding
carboxylic acid groups (or salt forms thereof); [0231] c)
conversion of all the carboxylic acid moieties into their
corresponding methyl esters; [0232] d) converting all the
azido-groups into the corresponding acylamino-groups; [0233] e)
selective removal of all R.sup.1' protecting groups; [0234] f)
sulfation of the resulting primary hydroxy groups; [0235] g)
selective removal of all R.sup.2' protecting groups and hydrolysis
of the methyl esters; [0236] h) sulfation of the resulting
secondary hydroxy groups or proceeding directly to step (i). [0237]
i) removal of the R.sup.3' protecting groups; and [0238] j)
conversion to the desired cationic salt form of the final
product.
[0239] Synthesis of a 4-Methoxyphenyl Glycoside Dodecasaccharide of
the Invention
[0240] A dodecasaccharide compound of the invention is synthesised
from five neutral disaccharide building blocks of general formula
(A) and/or (B), respectively, selected independently for the
reducing terminal and internal disaccharide units, and one of (C)
or (D) selected independently for the non-reducing terminal
disaccharide unit.
[0241] Thus, the decasaccharide (M) is prepared by: [0242] a)
synthesis of an octasaccharide acceptor by selective removal of the
protecting group R.sup.4' from octasaccharide derivative (K) to
generate a free secondary hydroxy group; [0243] b) coupling of the
donor (F) with this octasaccharide acceptor.
##STR00027##
[0244] Then dodecasaccharide (N) is prepared by: [0245] a)
synthesis of an decasaccharide acceptor by selective removal of the
protecting group R.sup.4' from octasaccharide derivative (M) to
generate a free secondary hydroxy group; [0246] b) coupling of the
donor (I) with this octasaccharide acceptor.
##STR00028##
[0247] where R.sup.16' and R.sup.17' are independently chosen from
hydrogen and CH.sub.2OR.sup.7' with the proviso that when one is
hydrogen, the other is CH.sub.2OR.sup.7'.
[0248] Finally, the dodecasaccharide of the invention is prepared
by: [0249] a) selective removal of all protecting groups R.sup.7'
from the neutral decasaccharide derivative (N); [0250] b) oxidation
of all the resulting free primary hydroxy groups to the
corresponding carboxylic acid groups (or salt forms thereof);
[0251] c) conversion of all the carboxylic acid moieties into their
corresponding methyl esters; [0252] d) converting all the
azido-groups into the corresponding acylamino-groups; [0253] e)
selective removal of all R.sup.1' protecting groups; [0254] f)
sulfation of the resulting primary hydroxy groups; [0255] g)
selective removal of all R.sup.2' protecting groups and hydrolysis
of the methyl esters; [0256] h) sulfation of the resulting
secondary hydroxy groups or proceeding directly to step i). [0257]
i) removal of the R.sup.3' protecting groups; and [0258] j)
conversion to the desired cationic salt form of the final
product.
[0259] Synthesis of Alternative Compounds of the Invention
[0260] An octa- deca- or dodecasaccharide compound of the invention
having an optionally substituted alkyl, aralkyl or aryl aglycone is
synthesised by selective cleavage of the 4-methoxyphenyl residue
from C-1 of the reducing sugar moiety on a hexasaccharide
derivative (K) or the decasaccharide moiety (L) or the
dodecasaccharide moiety (N), respectively, and introducing a
suitable leaving group at C-1 of the reducing sugar and then
coupling the resulting oligoglycosyl donor with an optionally
substituted alkyl, aralkyl or aryl alcohol. The stereochemistry of
the newly formed anomeric centre is typically beta due to
neighbouring group participation by the adjacent R.sup.2' ester
protecting group in the glycosidation reaction.
[0261] In an alternative approach, an octa- or deca- or
dodecasaccharide compound of the invention having an optionally
substituted alkyl, aralkyl or aryl aglycone is synthesised by
substituting the disaccharide building block (P) for the
disaccharide building block (E) that provides the reducing sugar
disaccharide moiety in a octa- deca- or dodecasaccharide of the
invention in the processes outlined above (under "synthesis of a
4-methoxyphenyl glycoside octasaccharide of the invention",
"synthesis of a 4-methoxyphenyl glycoside decasaccharide of the
invention" and "synthesis of a 4-methoxyphenyl glycoside
dodecasaccharide of the invention").
##STR00029##
[0262] where R.sup.A' and R.sup.B' are chosen from an optionally
substituted alkyloxy, aralkyloxy or aryloxy group and hydrogen,
with the proviso that when one is hydrogen, the other is an
optionally substituted alkyloxy, aralkyloxy or aryloxy group.
[0263] A disaccharide building block (P) is conveniently
synthesised by reaction of the glycosyl donor (F) with the desired
optionally substituted alkyl, aralkyl or aryl alcohol. As the
R.sup.2' is a ester protecting group, the product is usually the
beta-anomer (in which R.sup.A' is hydrogen).
[0264] Synthesis of the Disaccharide Building Blocks
[0265] The neutral disaccharide building blocks of general formula
(A) and (B) are synthesised by coupling a glycosyl donor of formula
(Q) with a monosaccharide acceptor of formula (S) or (T),
respectively.
[0266] The neutral disaccharide building blocks of general formula
(C) and (D) are synthesised by coupling a glycosyl donor of formula
(R) with a monosaccharide acceptor of formula (S) or (T),
respectively.
##STR00030##
[0267] The required 1,2-cis-configuration at C-1 and C-2 of the
non-reducing sugar residue in the disaccharide building blocks (A),
(B), (C) and (D) is attained by suitable choice of leaving group X
and glycosidation reagent. Suitable choices of X and glycosidation
reagent are X=beta-(methylthio)-, beta-(phenylthio)- or
beta-(4-methylthio)-, with the combination of N-iodosuccinimide and
silver triflate as glycosidation reagents.
[0268] Typical Glycosidation Reaction Conditions
[0269] For the glycosidation reactions in which a disaccharide (A),
(B), (C) or (D), a tetrasaccharide (G), a hexasaccharide (H), an
octasaccharide (J) or (K), or a decasaccharide (L) or (M), or a
dodecasaccharide (N), is synthesised, the donor is conveniently
used in a molar ratio to the acceptor in the range 1.0 to 1.5
equivalents, preferably in a range between 1.05 and 1.5
equivalents, preferably 1.2 to 1.3 equivalents, preferably 1.3
equivalents.
[0270] Conveniently the glycosyl donor is a glycosyl
trichloroacetimidate. Reactions with acceptor alcohols are
conveniently carried out in anhydrous toluene (40 mL per mmol
acceptor) at between -10 and 40.degree. C., in the presence of
powdered molecular sieves (4 .ANG.) and trimethylsilyl
trifluoromethanesulfonate (0.3 eq).
[0271] Abbreviations
[0272] NMR Nuclear magnetic resonance
[0273] TLC Thin layer chromatography
[0274] RT Room temperature
[0275] DCM Dichloromethane
[0276] Ac Acetyl
[0277] AcOH Acetic acid
[0278] BAIB Bis(acetoxy)iodobenzen
[0279] Bn Benzyl
[0280] Bz Benzoyl
[0281] CAN Ceric ammonium (IV) nitrate
[0282] ClAc Chloroacetyl
[0283] DABCO 1,4-Diazabicyclo[2.2.2]octane
[0284] DMAP 4-N,N-Dimethylaminopyridine
[0285] DMF N,N-Dimethylformamide
[0286] ESI Electrospray ionization
[0287] EtOAc Ethyl acetate
[0288] Fmoc Fluorenylmethoxycarbonyl
[0289] Fmoc-Cl Fluorenylmethoxycarbonyl chloride
[0290] HSQC Heteronuclear single quantum correlation
[0291] HRMS High resolution mass spectrum
[0292] gl.AcOH Glacial acetic acid
[0293] MeOAc Methoxyacetyl
[0294] MeOH Methanol
[0295] MS Mass spectrum
[0296] NBS N-Bromosuccinimide
[0297] NI N-Iodosuccinimide
[0298] TEMPO 2,2,6,6-Tetramethyl-1-piperidinyloxyl
[0299] TFA Trifluoroacetic acid
[0300] THF Tetrahydrofuran
[0301] TMS-diazomethane Trimethylsilylmethyl diazomethane
EXAMPLES
[0302] The following examples further illustrate the invention. It
is to be appreciated that the invention is not limited to the
examples.
Example 1
Synthesis of Compounds
##STR00031##
[0304] Preparation of 1
[0305] Methyl 2-azido-2-deoxy-1-thio-.beta.-D-glucopyranoside
(Pozsgay, V.; Glaudemans, C. P. J.; Robbins, J. B.; Schneerson, R.
Tetrahedron 1992, 48, 10249-10264) (20 g, 85 mmol) is dissolved in
200 mL dry pyridine. Trityl chloride (30 g, 108 mmol) is added and
the mixture stirred overnight. More trityl chloride (2 g) is added
and stirring continued for 3 h until TLC (petroleum ether/ethyl
acetate 1:1) confirms completion. The mixture is concentrated in
vacuo, coevaporated with toluene (2.times.150 mL), washed with
CuSO4, water, brine, concentrated to dryness and coevaporated with
DCM to an off-white foam. This is dissolved in dry DMF (100 mL),
cooled in an ice-bath and benzyl bromide (30.3 ml, 255 mmol) is
added, followed by sodium hydride 60% (11.22 g, 281 mmol) in
portions, the ice-bath is removed after addition of the first 2 g
of NaH. After stirring at room temperature for 1 h, the reaction is
again cooled in an ice-bath and quenched by addition of ethanol,
then diluted with 500 mL toluene, washed with water (3.times.1000
mL), brine (300 mL) and dried over magnesium sulfate. Solvents are
evaporated and the resulting syrup is coevaporated with
dichloromethane, the residue is taken up in toluene and crytallised
by addition of methanol. The solids are collected, washed with
methanol and dried under vacuum to give the
benzyl/trityl-derivative 1 in 94% yield (52.3 g, 80 mmol) as a
white powder. .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 143.9,
137.8, 137.6, 129.1, 128.8. 128.6, 128.3, 128.2, 128.1, 127.9,
127.8, 127.1, 86.5, 85.0, 84.0, 79.1, 77.8, 76.0, 75.0, 65.8, 62.4,
11.9
[0306] Preparation of 2
[0307] Tritylated compound 1 (55.3 g, 84 mmol) is combined with
acetic acid (300 mL), water (80 mL) is added and the mixture heated
to 80.degree. C. for 8 h. The reaction mixture is concentrated and
the product crystallises from ethyl acetate (100 mL) by addition of
petroleum ether in 80% yield (27.8 g, colourless crystals). The
obtained alcohol is dissolved in pyridine/acetic anhydride 2:1 (300
mL) and stirred for 6 h at ambient temperature, subsequently the
solvents are evaporated to a syrup, which crystallises on standing
and is triturated with petrol (450 mL) for 3 d. Solids are
collected and dried under vacuum to give 2 (27.1 g, 65.2 mmol) as a
white powder. .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6,
137.6, 137.4, 128.6, 128.5, 128.4, 128.1, 128.0, 127.8, 85.1, 84.5,
76.8, 65.7, 63.00, 20.8, 12.5
[0308] Preparation of 3
[0309] Methyl
2-azido-4,6-di-O-benzylidene-2-deoxy-1-thio-.beta.-D-glucopyranoside
(Rajaratnam, P.; Gupta, P.; Katavic, P.; Kuipers, K; Huyh, N.;
Ryan, S.; Falzun, T.; Tometzki, G. B.; Bornaghi, L.; Le Thanh, G.;
Abbenante, G.; Liu, L.; Meutermans, W.; Wimmer, N.; West, M. L.
Aust. J. Chem. 2010, 63, 693-699. Dekany, G.; Alchemia Pty. Ltd.,
US Patent Publication, U.S. Pat. No. 6,953,850 B1, 2005) (62.1 g,
192 mmol) is dissolved in dry DMF (150 mL), benzyl bromide (27.4
ml, 230 mmol) is added and cooled in an ice bath. Then sodium
hydride 60% (9.99 g, 250 mmol) is added portion-wise and the
reaction stirred at room temperature until TLC shows full
conversion to a less polar product. The reaction is quenched with
MeOH, then evaporated under high vacuum. Chloroform is added to the
residue and washed three times with water and dried over magnesium
sulfate to give a solid residue. Then diethyl ether is added to
dissolve a part of the solids and precipitated with petroleum
ether. Solids are collected and washed with petroleum ether to
yield an off-white solid (56.7 g) Evaporation of the mother liquor,
followed by trituration with ether/petroleum ether gives more
yellow solid, (12.67 g). Total of benzyl ether 3 68.37 g, 86%.
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 137.7, 137.2, 129.1,
128.5, 128.4, 128.3, 128.0, 126.0, 101.3, 85.2, 81.6, 80.9, 75.1,
70.5, 68.5, 65.3, 12.7
[0310] Preparation of 4
[0311] Dioxane (125 mL) and methanol (400 mL) are added to benzyl
ether 3 (68.3 g, 165 mmol) and the mixture is stirred at room
temperature. Acetyl chloride (8 mL, 113 mmol) is added and the
reaction mixture is stirred at room temperature for 1.5 h, until
TLC shows full conversion. The reaction is neutralized with
Amberlyst A26 (OH--) resin, filtered and the filtrate evaporated to
dryness. Silica chromatography (EtOAc/Hex 1:2-3:2) gives a pale
yellow syrup that crystallizes: methyl
2-azido-3-O-benzyl-2-deoxy-1-thio-.beta.-D-glucopyranoside 4 49.14
g, 91%. .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 137.9, 128.7,
128.5, 128.3, 128.1, 127.8, 84.9, 84.5, 79.5, 75.4, 70.1, 65.2,
62.0, 12.7
[0312] Preparation of 5
[0313] Compound 4 (47.9 g, 147 mmol) is dissolved in dry
dichloromethane (590 mL), Pyridine (71.4 ml, 883 mmol) is added and
cooled to -75.degree. C. Acetyl chloride (10.47 ml, 147 mmol) is
added slowly and the reaction is allowed to warm up to room
temperature over night. TLC confirms complete conversion and the
reaction mixture is washed with 10% HCl, aq. sodium bicarbonate and
brine (400 mL resp.), dried over magnesium sulfate, concentrated
and dissolved in hot toluene (100 ml). The product crystallises in
the fridge (4.degree. C.), is collected by filtration, washed with
petroleum ether and dried under oil-pump vacuum: methyl
6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-1-thio-.beta.-D-glucopyranoside
5 (50 g, 92% yield). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
171.7, 137.8, 128.7, 128.6, 128.2, 127.9, 127.7, 84.7, 84.2, 77.9,
75.5, 70.1, 65.2, 63.2, 20.8, 12.5
[0314] Preparation of 6
[0315] DMAP (700 mg, 5.73 mmol) is added to a solution of 5 (50 g,
136 mmol) in anhydrous pyridine (100 ml) and cooled in an ice-bath.
Fmoc-Cl (141 g, 544 mmol) is added and the ice-bath is removed
after 15 min. The resulting suspension is stirred at room
temperature for 1.5 h. The solvents are evaporated and the residue
is purified by repeated silica chromatography (toluene/ethyl
acetate 4%); clean fractions are combined and co-evaporated with
dichloromethane to form a foam: methyl
6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-4-O-(9-fluorenylmethyloxycarbonyl)--
1-thio-.beta.-D-glucopyranoside 6 (69 g, 117 mmol, 86% yield).
R.sub.f=0.15 (toluene/ethyl acetate 19:1), .sup.13C-NMR (125 MHz,
CDCl.sub.3) .delta. 170.6. 154.2, 143.2, 143.0, 141.3, 137.1,
128.4, 128.0, 127.2, 125.1, 124.9, 120.1, 84.6, 82.1, 75.7, 75.5,
74.3, 70.3, 65.1, 62.3, 46.8, 20.7, 12.4
[0316] Ido-Configured Monosaccharide Building Block Synthesis
01
[0317] Reaction of a known tetra-benzoate (Barroca, N.; Jacquinet,
J.-C. 2000, Carbohydr. Res., 329, 667-679) with 4-methoxyphenol and
subsequent Zemplen deacetylation affords a triol 7.
Isopropylidenation gives the compound 8. Benzylation afforded a
benzoate 9. Acidic hydrolysis gives a diol 10. Selective
chloroacetylation furnishes a mono-chloroacetate acceptor 11.
##STR00032##
[0318] Synthesis of 7
[0319] A solution of tetra-benzoate (Barroca, N.; Jacquinet, J.-C.
2000, Carbohydr. Res., 329, 667-679) (56.2 g, 82 mmol) in dry
dichloromethane (400 mL) is treated with 4-methoxyphenol (2 equiv.,
20.3 g, 164 mmol) and boron trifluoride diethyl etherate (0.5
equiv., 5 ml, 40.9 mmol) and stirred at room temperature for 2
hours. Then the reaction mixture is washed with saturated aq.
sodium bicarbonate twice, dried and concentrated. The residue is
dissolved in dry methanol (500 mL) and treated with 25% solution of
sodium methoxide (15 mL) and stirred at room temperature for 18
hours. The reaction mixture is neutralized with ion-exchange resin
(Amberlyst-H.sup.+) and the resin is filtered off. The residue is
purified by silica gel chromatography (EtOAc:petroleum ether, 1:2)
to furnish the triol 7 as a syrup: 25.0 g (81%), R.sub.f=0.15
(EtOAc:petroleum ether, 1:1). HRMS (ESI) calcd for
C.sub.20H.sub.24O.sub.7Na(M+Na).sup.+ m/z 399.142, found 399.1421.
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 7.39-7.23 (m, 5 H), 6.99
(d, J=9.1 Hz, 2H), 6.82 (d, J=9.2 Hz, 2H), 5.53 (d, J=4.5 Hz, 1H),
4.79 (d, J=12.4 Hz, 1H), 4.58 (d, J=11.9 Hz, 1H), 4.23 (d, J=8.8
Hz, 1H), 4.07 (dd, J=4.0, J=12.1, 1H), 3.89-3.83 (m, 3H), 3.81 (d,
J=9.5, 1H), 3.74 (s, 3H); .sup.13C-NMR (CDCl.sub.3) .delta. 154.9,
150.7, 138.0, 133.3, 130.1, 129.9, 128.5, 127.8, 127.6, 123.2,
117.9, 114.7, 100.4, 75.1, 71.7, 70.8, 66.1, 65.1, 60.5, 55.7.
[0320] Synthesis of 8
[0321] A solution of 7 (17 g, 45.2 mmol) in dry DMF (150 mL) is
treated with 2,2-dimethoxypropane (100 mL) and p-toluenesulfonic
acid monohydrate (100 mg) and stirred at room temperature for 5
hours. Then the reaction mixture is diluted with ethyl acetate,
washed with saturated aq. sodium bicarbonate and water, dried and
concentrated. The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:2) to afford the isopropylidene
derivative 8 as a syrup: 17.9 g, 43 mmol (95%), R.sub.f=0.75
(EtOAc:petroleum ether, 1:1). HRMS (ESI) calcd for
C.sub.23H.sub.28O.sub.7Na(M+Na).sup.+ m/z 439.1733, found 439.1725.
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 7.43-7.29 (m, 5 H), 7.04
(d, J=9.2 Hz, 2H), 6.82 (d, J=9.1 Hz, 2H), 5.59 (d, J=5.3 Hz, 1H),
4.8 (d, J=11.8 Hz, 1H), 4.62 (d, J=11.1 Hz, 1H), 4.08 (d, J=8.5 Hz,
1H), 4.03-3.97 (m, 3H), 3.81 (dd, J=4.2, J=12.3, 1H), 3.75 (s, 3H),
3.61 (d, J=11.2 Hz, 1H), 1.47 (s, 3H), 1.44 (s, 3H); .sup.13C-NMR
(125 MHz, CDCl.sub.3) .delta. 154.7, 150.9, 137.9, 128.5, 127.9,
127.7, 117.4, 114.6, 99.6, 99.3, 74.9, 71.7, 68.2, 65.2, 63.1,
60.5, 60.4, 55.7, 29.3, 18.5.
[0322] Synthesis of 9
[0323] A solution of 8 (17 g, 40.8 mmol) in dry dichloromethane
(100 mL) and dry pyridine (50 mL) is treated with benzoyl chloride
(2 equiv., 9.48 mL, 82 mmol) at 0.degree. C. The reaction mixture
is stirred at 0.degree. C. for 30 min followed by stirring at room
temperature for 5 hours. Then the mixture is diluted with ethyl
acetate and washed with saturated aq. sodium bicarbonate and water,
dried and concentrated. The residue is purified by silica gel
chromatography (EtOAc:petroleum ether, 1:3) to afford the benzoate
9 as a syrup: 20.0 g, 38.4 mmol (95%), R.sub.f=0.4 (EtOAc:petroleum
ether, 1:2). HRMS (ESI) calcd for
C.sub.30H.sub.32O.sub.8Na(M+Na).sup.+ m/z 543.1995, found 543.1985.
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.14 (d, J=7.6 Hz, 2H),
7.56-7.52 (m, 1 H), 7.43-7.38 (m, 4 H), 7.34-7.28 (m, 3 H), 7.05
(d, J=9.2 Hz, 2H), 6.81 (d, J=9.1 Hz, 2H), 5.68 (d, J=6.1 Hz, 1H),
5.49 (dd, J=2.5 Hz, J=6.2 Hz, 1H), 4.94 (d, J=11.4 Hz, 1H), 4.72
(d, J=11.5 Hz, 1H), 4.12-4.04 (m, 3H), 3.91 (dd, J=4.3, J=13.1,
1H), 3.82 (t, J=7.2, 1H), 3.74 (s, 3H), 3.61 (d, J=11.2 Hz, 1H),
1.49 (s, 3H), 1.45 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 165.5, 154.9, 150.7, 137.9, 133.5, 133.3, 130.1, 130.0,
129.8, 129.5, 128.4, 128.3, 127.8, 117.6, 114.6, 98.75, 97.55,
75.1, 72.1, 67.7, 67.1, 62.7, 61.1, 60.4, 55.7, 28.9, 18.9.
[0324] Synthesis of 10
[0325] A solution of 9 (20 g, 38.4 mmol) in gl.AcOH (160 mL) and
water (80 mL) is stirred at 80.degree. C. for 1 hour. Then the
solvents are removed in vacuo and the residue is purified by silica
gel chromatography (EtOAc:petroleum ether, 1:1) to give the diol 10
as a foam, 16.0 g, 33.3 mmol (87%), R.sub.f=0.15 (EtOAc:petroleum
ether, 1:2). HRMS (ESI) calcd for
C.sub.27H.sub.28O.sub.8Na(M+Na).sup.+ m/z 503.1682, found 503.1689.
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.04 (d, J=7.6 Hz, 2H),
7.61-7.56 (m, 1 H), 7.47-7.39 (m, 4 H), 7.36-7.27 (m, 3 H), 7.05
(d, J=9.2 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.57 (bs, 1H), 5.47-5.45
(m, 1H), 4.93 (d, J=12.0 Hz, 1H), 4.70 (d, J=11.5 Hz, 1H),
4.47-4.44 (m, 1H), 3.94-3.88 (m, 2H), 3.85-3.80 (m, 1H), 3.75 (s,
3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.2, 165.1,
155.2, 150.4, 137.8, 133.7, 129.8, 129.2, 128.7, 128.5, 127.9,
127.7, 118.3, 114.7, 97.9, 75.2, 72.0, 68.4, 68.1, 67.9, 63.2,
60.4, 55.6.
[0326] Synthesis of 11
[0327] A solution of 10 (15 g, 31.2 mmol) in dry dichloromethane
(100 mL) and dry pyridine (20 mL) is treated with a solution of
chloroacetyl chloride (0.85 equiv., 2.11 mL, 26.5 mmol) in dry
dichloromethane (5 mL) dropwise at -78.degree. C. The reaction
mixture is stirred at -78.degree. C. for 45 min, allowed to warm up
to 0.degree. C. and quenched with water (5 mL). Then the mixture is
diluted with dichloromethane, washed with saturated aq. sodium
bicarbonate and water, dried and concentrated. The residue is
purified by silica gel chromatography (EtOAc:petroleum ether, 1:3)
to afford the chloroacetate 11 as a foam, 15.0 g, 26.9 mmol (86%),
R.sub.f=0.35 (EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.29H.sub.29O.sub.9ClNa(M+Na).sup.+ m/z 579.1398, found
579.1395. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.03 (d, J=7.5
Hz, 2H), 7.62-7.59 (m, 1 H), 7.49-7.41 (m, 3 H), 7.39-7.35 (m, 2
H), 7.33-7.29 (m, 1 H), 7.05 (d, J=10.0 Hz, 2H), 6.85 (d, J=8.9 Hz,
2H), 5.56 (bs, 1H), 5.47-5.45 (m, 1H), 4.95 (d, J=11.8 Hz, 1H),
4.70 (d, J=11.8 Hz, 1H), 4.66-4.63 (m, 2H), 4.55-4.51 (m, 1H),
4.35-4.31 (m, 1H), 3.93-3.91 (m, 1H), 3.85 (s, 2H); 3.78 (s, 3H);
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 166.9, 164.9, 155.3,
150.1, 137.6, 133.8, 129.8, 128.9, 128.8, 128.5, 127.9, 127.8,
118.3, 114.6, 97.3, 74.7, 72.1, 67.5, 67.1, 66.3, 65.2, 60.4, 55.7,
40.8.
##STR00033##
[0328] Preparation of 12
[0329] Pyridine (0.447 ml, 5.53 mmol) is added to p-methoxyphenyl
2-O-benzoyl-3-O-benzyl-.beta.-D-glucopyranoside (Karst, N.;
Jacquinet, J.-C. 2002, Eur. J. Org. Chem., 815-825) (0.443 g, 0.922
mmol) in dry dichloromethane (5 mL) and cooled to -75.degree. C.
Chloroacetyl chloride (0.074 ml, 0.922 mmol) (1.0 mL of a solution
of 0.74 mL in 9.3 mL of DCM) is added slowly and the reaction is
kept in cold bath for 1 h, then water (1 mL) is added and warmed to
RT. The reaction mixture is washed with water, dil. HCl, aq. sodium
bicarbonate and dried over magnesium sulfate. Evaporation of
solvents gives a white solid which recrystallises from
EtOAc/hexanes: alcohol 12, 0.41 g, 80%. .sup.13C-NMR (75 MHz,
CDCl.sub.3) .delta. 167.8, 165.6, 156.1, 151.6, 138.0, 133.7,
130.2, 130.0, 129.0, 128.9, 128.5, 119.2, 114.9, 101.3, 82.6, 75.1,
73.9, 73.8, 70.3, 65.1, 56.0, 41.1
##STR00034##
##STR00035##
[0330] General Procedure A (GPA): Disaccharide Building Block
Synthesis. Thioglycoside donor (1.5 eq) and alcohol (1.0 eq) are
dissolved in a mixture of anhydrous toluene and anhydrous
dichloromethane (25 mL per mmol acceptor) and cooled to -15.degree.
C. and powdered molecular sieves (4 .ANG.) are added. After 10 min
N-iodosuccinimide (1.7 eq) and silver trifluoromethanesulfonate
(0.4 eq) are added. The reaction mixture is allowed to warm up to
room temperature over 1 h. The mixture is diluted with ethyl
acetate and filtered through celite. The filtrate is washed with a
1:1 mixture of saturated aq. sodium bicarbonate and aq. thiosulfate
(30%), washed with saturated aq. sodium chloride, dried over
magnesium sulfate and concentrated. The residue is purified by
flash chromatography or crystallisation.
[0331] Synthesis of 13
[0332] Compound 13 is prepared from compound 12 and compound 6
according to general procedure A1: 11.2 g (71% crystalline
.alpha.-anomer), crystallised from hot toluene (60 mL) after
addition of petroleum ether. .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.5, 169.9, 165.1, 155.7, 154.2, 151.0, 143.2, 143.0,
141.3, 137.3, 137.1, 135.5, 129.6, 129.5, 129.0, 128.9, 128.6,
128.5, 128.4, 128.2, 128.0, 127.9, 127.7, 127.2, 125.3, 125.1,
124.9, 120.1, 118.7, 114.6, 100.0, 97.9, 82.5, 75.1, 74.8, 74.7,
74.1, 73.5, 72.4, 70.4, 68.8, 64.9, 62.6, 62.0, 55.7, 46.7, 40.6,
20.7
[0333] Synthesis of 14
[0334] Compound 14 is prepared from compound 12 and compound 2
according to general procedure A1: 1.11 g (79% .alpha.-anomer),
crystallised from toluene after addition of petroleum ether.
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.5, 166.9, 165.1,
155.7, 151.0, 137.5, 137.4, 137.2, 133.5, 129.8, 129.5, 129.1,
129.0, 128.8, 128.6, 128.5, 128.4, 128.1, 128.0, 127.8, 125.3,
118.7, 114.5, 100.1, 98.1, 82.7, 80.1, 79.5, 77.9, 75.5, 75.3,
74.9, 74.7, 74.2, 73.6, 72.4, 70.4, 64.9, 63.2, 62.6, 55.6, 40.4,
20.7
[0335] Synthesis of 15
[0336] Compound 15 is prepared from compound 2 and p-methoxyphenyl
2-O-benzoyl-3-O-benzyl-6-O-methoxyacetyl-.beta.-D-glucopyranoside
according to general procedure A1: 850 mg (79% .alpha.-anomer),
crystallised from toluene after addition of petroleum ether.
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.5, 169.9, 165.1,
155.7, 151.1, 137.6, 137.4, 137.2, 133.4, 129.8, 129.5, 129.0,
128.6, 128.5, 128.2, 128.1, 128.0, 127.8, 127.7, 125.3, 118.8,
114.5, 100.2, 98.0, 82.8, 80.1, 77.9, 75.5, 75.2, 74.5, 74.2, 73.7,
72.6, 70.3, 69.5, 63.5, 63.2, 62.6, 59.4, 55.6, 20.8
[0337] Synthesis of 16
[0338] Compound 16 is prepared from compound 11 and compound 6
according to general procedure A: 7.7 g (89%) crystalline
.alpha.-anomer, R.sub.f=0.45 (EtOAc:petroleum ether, 1:2).
Crystallised from hot ethyl acetate (80 mL) after addition of
petroleum ether (20%). HRMS (ESI) calcd for
C.sub.59H.sub.56ClN.sub.3O.sub.16Na(M+Na).sup.+ m/z 1120.3247,
found 1120.3246. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.18
(dd, J=3.7 Hz, J=9.9 Hz, 2 H), 7.79 (d, J=8.6 Hz, 2H), 7.61 (d,
J=8.6 Hz, 1H), 7.56 (d, J=7.5 Hz, 1H), 7.47-7.42 (m, 4H), 7.41-7.35
(m, 3H), 7.33-7.26 (m, 4H), 7.18-7.15 (m, 3H), 7.06 (d, J=9.1 Hz
2H), 7.03-6.99 (m, 3H), 6.86 (d, J=10.1 Hz 2H), 5.63 (d, J=4.1 Hz,
1H), 5.38 (d, J=3.9 Hz, 1H), 5.03 (d, J=12.1 Hz, 1H), 4.18 (d,
J=12.1 Hz, 2H), 4.77 (t, J=9.9 Hz, 1H), 4.66 (d, J=4.3 Hz, 1H),
4.61-4.58 (m, 3H), 4.56-4.52 (m, 1H), 4.51-4.47 (m, 1H), 4.33-4.27
(m, 1H), 4.25-4.20 (m, 1H), 4.19-4.15 (m, 1H), 4.10 (d, J=3.3 Hz,
1H), 4.09-4.05 (m, 1H), 4.01 (d, J=11.0 Hz, 3H), 3.85 (d, J=2.5,
2H), 3.77 (s, 3H), 3.72 (d, J=7.3 Hz, 1H), 3.65 (t, J=10.2 Hz, 1H),
3.36-3.33 (m, 1H), 2.01 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.5, 166.9, 165.6, 155.3, 154.2, 150.1, 143.2, 143.1,
141.3, 137.6, 137.1, 133.5, 129.9, 129.8, 128.7, 128.5, 128.2,
128.1, 128.0, 127.9, 127.8, 127.7, 127.2, 125.1, 124.9, 120.1,
118.3, 114.6, 98.9, 97.6, 78.3, 75.7, 75.0, 74.5, 72.4, 72.3, 70.3,
68.9, 68.1, 65.6, 65.0, 63.3, 62.1, 55.7, 46.7, 40.5, 20.6.
[0339] Synthesis of 17
[0340] Compound 17 is prepared from compound 11 and compound 2
according to general procedure A: . The residue is purified by
silica gel chromatography (EtOAc:petroleum ether, 1:3) to furnish
the disaccharide as a foam, 3.7 g (93%), .alpha.-anomer,
R.sub.f=0.45 (EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.51H.sub.52ClN.sub.3O.sub.14Na(M+Na).sup.+ m/z 988.3036, found
988.3043. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.16 (d, J=8.1
Hz, 2 H), 7.46 (d, J=7.9 Hz, 2 H), 7.42 (d, J=7.1 Hz, 2 H),
7.39-7.34 (m, 3H), 7.33-7.28 (m, 5H), 7.26-7.22 (m, 4H), 7.16 (dd,
J=7.5 Hz, J=12.3 Hz, 2H), 7.05 (d, J=10.1 Hz, 2H), 6.85 (d, J=9.3
Hz, 2H), 5.61 (d, J=4.0 Hz, 1H), 5.35 (d, J=3.7 Hz, 1H), 5.02 (d,
J=12.1 Hz, 1H), 4.81 (d, J=12.2 Hz, 2H), 4.75 (d, J=10.8 Hz, 2H),
4.64 (d, J=4.3 Hz, 2H), 4.59-4.55 (m, 3H), 4.53-4.51 (m, 1H),
4.50-4.48 (m, 1H), 4.35-4.30 (m, 1H), 4.29 (d, J=3.0 Hz, 1H), 4.29
(d, J=3.0 Hz, 1H), 4.27 (d, J=3.0 Hz, 1H), 4.21-4.17 (m, 1H), 4.13
(d, J=8.0 Hz, 2H), 4.10 (d, J=10.0 Hz, 2H), 3.94-3.91 (m, 1H), 3.84
(d, J=4.8, 2H), 3.77 (s, 3H), 3.72-3.69 (m, 1H), 3.65 (t, J=10.2
Hz, 1H), 3.39 (t, J=9.2 Hz, 1H), 3.31 (dd, J=6.5 Hz, J=12.9 Hz,
1H), 2.0 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.5,
166.8, 165.6, 155.2, 150.2, 137.6, 137.4, 137.3, 133.3, 129.9,
129.8, 128.7, 128.5, 128.4, 128.12, 128.1, 127.9, 127.96, 127.9,
127.7, 118.3, 114.5, 98.8, 97.5, 80.7, 77.8, 75.2, 75.1, 72.3,
70.4, 68.2, 65.8, 65.1, 63.8, 62.8, 55.7, 40.5, 20.7.
[0341] General Procedure B (GPB): Anomeric Deprotection. Ammonium
cerium (IV) nitrate (2 eq) is added to a solution of the starting
p-methoxyphenyl glycoside (1 eq) in acetonitrile/water 7:1 (12.5 mL
per mmol). The mixture is stirred at room temperature until TLC
(hexanes/ethyl acetate 1:1) indicates complete consumption of the
starting material, 0.5 to 3 d. The reaction mixture is diluted with
ethyl acetate, washed with water (twice), saturated aq. sodium
chloride, dried over magnesium sulfate and concentrated. The
residue is purified by flash chromatography to give the product as
a foam.
[0342] Synthesis of 18
[0343] Compound 18 is prepared from compound 13 following general
procedure B: 4.37 g, 90% (2 anomers), R.sub.f=0.2/0.25
(Toluene/EtOAc 5:1); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.5, 166.7, 165.7, 164.8, 161.0, 160.5, 154.1, 143.2, 143.0,
141.3, 137.8, 137.3, 137.2, 137.1, 133.8, 133.5, 129.9, 129.8,
129.4, 129.0, 128.6, 128.4, 128.3, 128.0, 127.9, 127.7, 127.3,
127.2, 125.1, 124.9, 120.1, 98.1, 97.9, 95.6, 90.2, 82.4, 79.4,
77.5, 76.4, 75.2, 74.8, 74.7. 74.3, 72.5, 70.4, 68.8, 68.7, 68.1,
64.9, 64.7, 62.7, 62.6, 61.9, 46.7, 40.7, 20.7
[0344] Synthesis of 19
[0345] Compound 19 is prepared from compound 14 following general
procedure B: 2.9 g, 72% (2 anomers), R.sub.f=0.4/0.5 (Toluene/EtOAc
7:3); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.5, 167.1,
166.7, 165.7, 137.9, 137.6, 137.3, 133.5, 129.9, 129.8, 129.4,
129.0, 128.6, 128.4, 128.3, 128.1, 128.0, 127.8, 127.6, 127.3,
98.3, 98.1, 90.2, 80.1, 79.5, 77.9, 76.5, 75.6, 75.2, 75.0, 74.8,
74.4, 70.4, 70.3, 68.3, 65.0, 64.8, 63.3, 62.5, 62.4, 40.7,
20.8
[0346] Synthesis of 20
[0347] Compound 20 is prepared from compound 16 following general
procedure B: to obtain a mixture of disaccharides as a foam, 6.0 g,
78% yield (2 anomers), TLC R.sub.f=0.25/0.3 (EtOAc:petroleum ether,
1:2). HRMS (ESI) calcd for
C.sub.52H.sub.50ClN.sub.3O.sub.15Na(M+Na).sup.+ m/z 1014.2828,
found 1014.283. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.18-8.12
(m, 4 H), 7.77 (d, J=7.5 Hz, 4 H), 7.59 (d, J=7.6 Hz, 2 H), 7.55
(d, J=7.8 Hz, 2 H), 7.44-7.35 (m, 8H), 7.35-7.30 (m, 8H), 7.29-7.26
(m, 6H), 7.24 (d, J=3.0 Hz, 4H), 7.17-7.14 (m, 4H), 6.99-6.55 (m,
4H), 5.30 (d, J=8.6 Hz, 2H), 5.26 (d, J=5.5 Hz, 2H), 4.91 (d,
J=11.3 Hz, 1H), 4.88 (d, J=12.1 Hz, 1H), 4.78 (d, J=11.4 Hz, 2H),
4.75 (d, J=9.0 Hz, 2H), 4.74-4.72 (m, 1H), 4.71 (dd, J=3.0 Hz,
J=6.1 Hz, 1H), 4.69 (d, J=9.0 Hz, 2H), 4.56-4.53 (m, 2H), 4.52-4.51
(m, 2H), 4.50-4.47 (m, 2H), 4.39-4.35 (m, 2H), 4.34-4.32 (m, 2H),
4.30-4.28 (m, 2H), 4.25-4.21 (m, 2H), 4.29-4.20 (m, 2H), 4.19-4.14
(m, 2H), 4.13 (bs, 2H), 4.10 (s, 2H), 4.09 (s, 2H), 4.09-4.0 (m,
1H), 3.93 (t, J=4.6 Hz, 1H), 3.89 (d, J=3.2 Hz, 2H), 3.84-3.82 (m,
1H), 3.81-3.80 (m, 1H), 3.64-3.60 (m, 1H), 3.54-3.52 (m, 1H),
3.49-3.44 (m, 1H), 3.31 (dd, J=3.9 Hz, J=13.8 Hz, 1H), 2.04 (s,
3H), 2.03 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.6, 170.5, 167.1, 167.0, 165.8, 165.7, 154.2, 143.2, 143.1,
141.4, 141.3, 137.0, 136.3, 133.6, 133.5, 129.9, 129.8, 129.7,
129.6, 128.9, 128.8, 128.7, 128.6, 128.4, 128.3, 128.2, 127.9,
127.8, 127.7, 127.3, 125.0, 124.8, 120.1, 114.7, 99.8, 99.6, 93.2,
92.1, 79.7, 78.3, 78.2, 76.8, 76.1, 75.8, 74.9, 74.8, 74.4, 73.6,
73.2, 73.1, 72.5, 72.2, 70.2, 69.3, 69.1, 69.0, 67.6, 65.2, 65.1,
64.3, 63.3, 63.2, 61.9, 60.4, 46.8, 40.7, 20.7.
[0348] Synthesis of 21
[0349] Compound 21 is prepared from compound 17 following general
procedure B: to obtain a mixture of anomers as a syrup 4.1 g, 80%
yield (2 anomers), R.sub.f=0.2/0.25 (EtOAc:petroleum ether, 1:2).
HRMS (ESI) calcd for
C.sub.44H.sub.46ClN.sub.3O.sub.13Na(M+Na).sup.+ m/z 882.2617, found
882.2619. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.16-8.11 (m, 4
H), 7.39-7.36 (m, 8 H), 7.35-7.31 (m, 8 H), 7.29-7.26 (m, 6 H),
7.25 (d, J=3.8 Hz, 10 H), 7.23-7.20 (m, 2H), 7.12-7.09 (m, 2H),
5.28 (d, J=9.6 Hz, 2H), 5.24 (dd, J=7.8 Hz, J=12.3 Hz, 2H),
5.08-5.05 (m, 4H), 4.90 (bs, 2H), 4.88 (d, J=3.4 Hz, 1H), 4.85 (bs,
1H), 4.78 (d, J=11.8 Hz, 2H), 4.75 (d, J=11.5 Hz, 2H), 4.71 (d,
J=10.8 Hz, 2H), 4.54-4.51 (m, 2H), 4.50-4.46 (m, 4H), 4.40-4.36 (m,
2H), 4.32-4.27 (m, 2H), 4.24-4.21 (m, 2H), 4.20-4.16 (m, 2H), 4.12
(bs, 1H), 4.105 (s, 2H), 4.101 (s, 2H), 3.93-3.87 (m, 2H),
3.62-3.61 (m, 1H), 3.54 (t, J=5.3 Hz, 1H), 3.50 (d, J=9.6 Hz, 1H),
3.47 (d, J=7.3 Hz, 1H), 3.41 (d, J=10.8 Hz, 1H), 3.36 (t, J=9.5 Hz,
1H), 3.28-3.24 (m, 1H), 2.04 (s, 3H), 2.02 (s, 3H); .sup.13C-NMR
(125 MHz, CDCl.sub.3) .delta. 170.6, 167.1, 167.0, 165.9, 165.8,
137.4, 137.3, 137.1, 136.3, 133.4, 133.3, 129.84, 129.8, 128.8,
128.7, 128.6, 128.5, 128.4, 128.3, 128.1, 127.9, 99.6, 99.5, 93.1,
92.0, 80.7, 80.6, 76.8, 75.6, 75.3, 75.2, 74.9, 73.6, 73.1, 72.3,
72.2, 70.5, 69.4, 67.8, 65.4, 65.2, 64.5, 63.8, 62.7, 60.4, 40.7,
20.7.
[0350] General Procedure C (GPC): Trichloroacetimidate Formation.
Hemi-acetal starting material (1 eq) is dissolved in
trichloroacetonitrile (20 eq) and the same volume of
dichloromethane. The mixture is cooled in an ice-bath and sodium
hydride (60% in mineral oil) (0.05 eq) is added. After 5 min the
ice-bath is removed and the reaction allowed to warm up to room
temperature and left until completion. The reaction mixture is
subjected to flash chromatography to yield the trichloroacetimidate
donor as an off-white foam.
[0351] Synthesis of 22
[0352] Compound 22 is prepared from compound 18 following general
procedure C: 3.8 g, 81% (2 anomers), R.sub.f=0.6/0.7 (Toluene/EtOAc
5:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.5, 170.4,
167.0, 165.3, 164.8, 161.0, 160.5, 154.2, 154.1, 143.2, 143.0,
141.3, 137.4, 137.1, 133.6, 129.8, 129.7, 129.2, 129.1, 129.0,
128.7, 128.5, 128.4, 128.3, 128.2, 127.9, 127.8, 127.7, 127.3,
127.2, 125.3, 125.1, 124.9, 120.1, 98.4, 98.2, 95.6, 93.2, 80.8,
80.1, 79.9, 77.4, 75.1, 75.0, 74.8, 74.6, 73.7, 72.9, 72.8, 68.9,
68.8, 64.8, 64.4, 62.7, 62.5, 61.9, 61.8, 46.7, 40.7, 40.6,
20.7
[0353] Synthesis of 23
[0354] Compound 23 is prepared from compound 19 following general
procedure C: 2.84 g, 85% (2 anomers), R.sub.f=0.45/0.5
(Toluene/EtOAc 9:1); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.5, 170.4, 167.0, 165.3, 164.8, 137.6, 137.2, 133.6, 133.5,
129.8, 129.7, 129.3, 129.1, 129.0, 128.6, 128.5, 128.4, 128.25,
128.2, 128.15, 128.1, 128.0, 127.8, 127.7, 127.4, 125.3, 98.7,
98.3, 95.7, 93.2, 81.4, 80.1, 79.9, 77.9, 77.8, 75.6, 75.5, 75.2,
75.0, 74.6, 74.5, 73.8, 73.0, 72.9, 64.8, 64.4, 63.3, 63.1, 62.6,
62.4, 40.7, 40.6, 20.8
[0355] Synthesis of 24
[0356] Compound 24 is prepared from compound 20 following general
procedure C: to obtain a mixture of anomers as a, foam 6.1 g, 82%
yield (2 anomers), R.sub.f=0.35/0.4 (EtOAc:petroleum ether, 1:2).
HRMS (ESI) calcd for
C.sub.54H.sub.50Cl.sub.4N.sub.4O.sub.15Na(M+Na).sup.+ m/z
1157.1924, found 1157.1929. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.70 (s, 1H), 8.59 (s, 1H), 8.18-8.15 (m, 2 H), 8.07 (dd,
J=7.4 Hz, J=9.6 Hz, 2 H), 7.78-7.75 (m, 2 H), 7.61-7.58 (m, 2 H),
7.61-7.58 (m, 4 H), 7.57-7.54 (m, 4 H), 7.49-7.46 (m, 2 H),
7.44-7.38 (m, 4 H), 7.37-7.34 (m, 4 H), 7.32-7.31 (m, 2 H),
7.31-7.27 (m, 2 H), 7.27-7.26 (m, 2 H), 7.25 (s, 4 H), 7.20-7.15
(m, 4H), 7.12 (dd, J=4.4 Hz, J=10.0 Hz, 2H), 7.0 (dd, J=6.4 Hz,
J=9.2 Hz, 2H), 6.50 (d, J=2.7 Hz, 1H), 6.45 (s, 1H), 5.41-5.39 (m,
1 H), 5.36 (t, J=2.1 Hz, 1H), 5.02 (d, J=2.9 Hz, 1H), 4.97 (d,
J=11.4 Hz, 1H), 4.89 (s, 2H), 4.87 (d, J=8.3 Hz, 1H), 4.83 (d,
J=9.8 Hz, 1H), 4.80 (d, J=9.8 Hz, 2H), 4.78-4.72 (m, 1H), 4.69 (d,
J=3.4 Hz, 1H), 4.68-4.64 (m, 2H), 4.63 (d, J=5.3 Hz, 2H), 4.60 (d,
J=7.6 Hz, 2H), 4.58 (d, J=6.0 Hz, 2H), 4.55 (d, J=4.3 Hz, 2H), 4.53
(d, J=3.2 Hz, 2H), 4.52 (d, J=3.7 Hz, 2H), 4.50 (d, J=2.5 Hz, 2H),
4.48 (d, J=3.9 Hz, 2H), 4.46-4.42 (m, 1H), 4.39-4.35 (m, 1H), 4.32
(d, J=3.4 Hz, 1H), 4.31-4.28 (m, 1H), 4.25-4.22 (m, 1H), 4.22-4.20
(m, 1H), 4.19-4.16 (m, 1H), 4.14-4.07 (m, 1H), 4.058 (s, 2H), 4.054
(s, 2H), 3.99 (d, J=10.8 Hz, 1H), 3.92 (d, J=10.8 Hz, 1H),
3.76-3.71 (m, 1H), 3.60 (t, J=9.8 Hz, 1H), 3.40 (t, J=5.3 Hz, 1H),
3.37 (dd, J=6.8 Hz, J=13.9 Hz, 1H), 2.05 (s, 3H), 2.04 (s, 3H);
.sup.13C-NMR (CDCl.sub.3) .delta. 171.1, 170.5, 167.0, 166.9,
165.7, 165.4, 160.6, 160.5, 154.2, 143.2, 143.1, 141.3, 137.3,
137.2, 137.0, 133.6, 133.5, 129.9, 129.8, 129.5, 129.4, 128.8,
128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8,
127.7, 127.3, 125.0, 124.9, 124.8, 120.1, 99.5, 99.3, 95.5, 94.9,
78.3, 77.9, 76.8, 75.6, 75.4, 75.1, 74.9, 74.6, 74.5, 73.5, 72.4,
71.6, 70.4, 70.2, 69.9, 69.0, 68.8, 67.0, 66.4, 65.4, 64.9, 63.3,
63.1, 62.1, 62.0, 60.4, 46.8, 40.6, 40.5, 20.7.
[0357] Synthesis of 25
[0358] Compound 25 is prepared from compound 21 following general
procedure C: to obtain a mixture of anomers as a foam, 4.4 g, 94%
yield (2 anomers), TLC, R.sub.f=0.45/0.5, (EtOAc:petroleum ether,
1:2). HRMS (ESI) calcd for
C.sub.46H.sub.46Cl.sub.4N.sub.4O.sub.13Na(M+Na).sup.+ m/z
1025.1713, found 1025.1718. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.68 (s, 1H), 8.57 (s, 1H), 8.16 (dd, J=7.3 Hz, J=9.5 Hz, 2
H), 8.05 (dd, J=7.1 Hz, J=9.4 Hz, 2 H), 7.49-7.44 (m, 2 H),
7.42-7.38 (m, 4 H), 7.37-7.30 (m, 10 H), 7.29-7.23 (m, 12 H),
7.14-7.12 (m, 6 H), 6.50 (d, J=2.8 Hz, 1H), 6.43 (s, 1H), 5.38 (dd,
J=4.5 Hz, J=10.2 Hz, 1 H), 5.35-5.33 (m, 1H), 5.28 (s, 1H), 5.04
(d, J=3.4 Hz, 1H), 4.96 (d, J=11.5 Hz, 1H), 4.87 (s, 1H), 4.80 (d,
J=10.2 Hz, 1H), 4.76 (t, J=10.2 Hz, 1H), 4.68 (d, J=3.4 Hz, 1H),
4.66-4.63 (m, 1H), 4.62 (d, J=6.2 Hz, 1H), 4.60-4.56 (m, 1H),
4.54-53 (m, 1H), 4.52-4.49 (m, 2H), 4.50-4.44 (m, 1H), 4.43 (d,
J=5.9 Hz, 2H), 4.40 (d, J=4.2 Hz, 2H), 4.38 (t, J=2.5 Hz, 1H), 4.36
(d, J=7.0 Hz, 2H), 4.32 (t, J=2.5 Hz, 1H), 4.30-4.28 (m, 1H), 4.27
(d, J=10.8 Hz, 1H), 4.23-4.17 (m, 1H), 4.15-4.13 (m, 1H), 4.12 (s,
2H), 4.11 (s, 2H), 4.06 (s, 1H), 4.04 (d, J=2.4 Hz, 1H), 4.04-4.02
(m, 1H), 4.0-3.97 (m, 1H), 3.96-3.93 (m, 1H), 3.73 (t, J=9.8 Hz,
1H), 3.61 (d, J=9.8 Hz, 1 H), 3.59 (d, J=9.8 Hz, 1 H), 3.46-3.41
(m, 1H), 3.40-3.36 (m, 1H), 3.35 (d, J=3.8 Hz, 1H), 3.33 (dd, J=3.3
Hz, J=6.1 Hz, 1H), 2.03 (s, 6H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 171.1, 170.6, 167.0, 166.9, 165.7, 165.5, 163.4, 160.6,
160.5, 137.5, 137.4, 137.3, 137.2, 133.4, 129.9, 129.8, 129.6,
129.3, 128.7, 128.6, 128.5, 128.4, 128.1, 128.0, 127.9, 99.4, 99.2,
95.5, 94.9, 80.7, 80.4, 77.8, 77.0, 76.8, 75.4, 75.3, 75.2, 75.0,
74.6, 74.5, 73.6, 72.3, 71.5, 70.6, 70.3, 67.2, 66.5, 65.7, 65.0,
63.9, 63.6, 62.7, 60.4, 40.7, 40.6, 21.0, 20.8.
[0359] General Procedure D (GPD): Fmoc Deprotection. The
appropriate Fmoc derivative is dissolved in a mixture of
dichloromethane and triethylamine 4:1 (v/v, 20 mL per mmol) and
left at ambient temperature until completion, usually 3 to 5 h.
Subsequently the mixture is diluted with dichloromethane, washed
with diluted aq. HCl, water and saturated aq. sodium chloride,
dried over magnesium sulfate and concentrated. The residue is
purified by flash chromatography to afford the desired alcohols as
foams.
##STR00036##
[0360] General Procedure E (GPE): Chain Extension Glycosylation. A
solution of the trichloroacetimidate donor (1.3 eq) and the
glycosyl acceptor alcohol (1 eq) in anhydrous toluene (40 mL per
mmol acceptor) is cooled to reaction temperature (between -10 and
-20.degree. C.), powdered molecular sieves (4 .ANG.) are added and
the suspension stirred at the temperature. After 15 min,
trimethylsilyl trifluoromethanesulfonate (0.3 eq) is added and the
reaction mixture stirred at reaction temperature until TLC
(toluene/ethyl acetate 4:1) indicated completion. The mixture is
diluted with ethyl acetate and filtered through celite into aq.
sodium bicarbonate, the organic layer is washed with water and
saturated aq. sodium chloride, dried over magnesium sulfate and
concentrated. The residue is purified by flash chromatography to
yield the fully protected oligosaccharides.
[0361] Synthesis of 26
[0362] Compound 26 is prepared from compound 16 following general
procedure D: 2.55 g, 88% yield, TLC, R.sub.f=0.2 (EtOAc:petroleum
ether, 1:2). HRMS (ESI) calcd for
C.sub.44H.sub.46ClN.sub.3O.sub.14Na(M+Na).sup.+ m/z 898.2566, found
898.2555. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.18 (dd, J=7.1
Hz, J=9.4 Hz, 2 H), 7.52-7.48 (m, 3 H), 7.47-7.43 (m, 3 H),
7.38-7.35 (m, 2 H), 7.34-7.27 (m, 3 H), 7.24-7.13 (m, 2 H), 7.06
(d, J=9.5 Hz, 2 H), 6.85 (d, J=9.5 Hz, 2 H), 5.62 (s, 1H), 5.36 (s,
1H), 5.02 (d, J=10.3 Hz, 1H), 4.81 (d, J=10.8 Hz, 1H), 4.67 (d,
J=3.6 Hz, 1H), 4.61-4.58 (m, 1H), 4.53 (d, J=9.1 Hz, 1H), 4.51 (dd,
J=6.7 Hz, J=10.3 Hz, 1H), 4.49 (d, J=4.3 Hz, 1H), 4.39 (d, J=10.6
Hz, 1H), 4.33 (d, J=3.5 Hz, 1H), 4.33 (d, J=4.4 Hz, 1H), 4.22-4.20
(m, 1H), 4.17 (d, J=2.2 Hz, 1H), 3.87 (d, J=2.0 Hz, 2H), 3.78 (s,
3H), 3.73 (t, J=2.8 Hz, 1H), 3.50 (d, J=8.2 Hz, 1H), 3.37 (d, J=8.9
Hz, 1H), 3.36-3.31 (m, 1H), 3.26 (dd, J=6.1 Hz, J=13.6 Hz, 1H),
2.05 (s, 3H); .sup.13C-NMR (CDCl.sub.3) .delta. 171.8, 166.9,
165.7, 155.3, 150.2, 137.7, 137.6, 133.4, 129.9, 129.8, 129.0,
128.7, 128.6, 128.4, 128.2, 128.13, 128.1, 128.0, 125.3, 118.3,
114.6, 98.9, 97.6, 80.1, 75.2, 75.0, 72.4, 72.36, 72.35, 70.5,
68.3, 65.8, 65.1, 63.3, 62.9, 55.7, 40.5, 20.7.
[0363] Synthesis of 27
[0364] Compound 27 is prepared from compound 13 following general
procedure D: 4.36 g, 91%, R.sub.f=0.3 (Toluene/EtOAc 4:1).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.9, 167.0, 165.1,
155.7, 151.0, 137.8, 137.3, 133.5, 129.8, 129.5, 128.7, 128.6,
128.4, 128.2, 127.8, 127.7, 118.7, 114.5, 100.1, 98.2, 82.8, 79.2,
75.4, 74.5, 74.2, 73.6, 72.5, 71.4, 70.7, 64.9, 62.9, 62.7, 55.6,
40.6, 20.8
[0365] Synthesis of 28
[0366] Compound 28 is prepared from compound 26 and compound 24
following general procedure E: 450 mg, 95% yield (beta), TLC,
R.sub.f=0.35 (EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.96H.sub.94Cl.sub.2N.sub.6O.sub.28Na(M+Na).sup.+ m/z
1871.5391, found 1871.5383. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.17 (dd, J=6.8 Hz, J=9.4 Hz, 2 H), 8.11 (dd, J=6.8 Hz,
J=9.8 Hz, 2 H), 7.77 (d, J=7.4 Hz, 2H), 7.60 (d, J=7.4 Hz, 2H),
7.55 (d, J=7.5 Hz, 1H), 7.47-7.43 (m, 4H), 7.43-7.39 (m, 3H),
7.39-7.36 (m, 3H), 7.35-7.32 (m, 4H), 7.32-7.31 (m, 4H), 7.30-7.23
(m, 4H), 7.20-7.15 (m, 4H), 7.08-7.05 (m, 4H), 7.06 (d, J=9.5 Hz,
2H), 6.85 (d, J=8.9 Hz, 2H), 5.64 (bd, 1H), 5.35 (t, J=2.4 Hz, 1H),
5.15 (t, J=3.9 Hz, 1H), 5.12 (d, J=3.5 Hz, 1H), 5.02 (d, J=11.8 Hz,
1H), 4.87 (d, J=11.5 Hz, 1H), 4.81 (d, J=11.3 Hz, 1H), 4.76 (d,
J=2.5 Hz, 1H), 4.73 (t, J=4.6 Hz, 1H), 4.65 (d, J=3.7 Hz, 1H),
4.59-4.55 (m, 1H), 4.51-4.48 (m, 2H), 4.48-4.44 (m, 2H), 4.38-4.34
(m, 3H), 4.33-4.29 (m, 4H), 4.28-4.25 (m, 2H), 4.24-4.23 (m, 1H),
4.22-4.20 (m, 2H), 4.19-4.15 (m, 2H), 4.09-4.07 (m, 2H), 4.05-4.03
(m, 1H), 4.03-4.01 (m, 1H), 4.0 (d, J=10.2 Hz, 1H), 3.96-3.94 (m,
1H), 3.92 (d, J=3.0 Hz, 2H), 3.83 (d, J=3.6, 2H), 3.77 (s, 3H),
3.72-3.68 (m, 1H), 3.68-3.65 (m, 1H), 3.60 (t, J=9.8, 1H),
3.38-3.28 (m, 1H), 2.03 (s, 3H), 2.02 (s, 3H); .sup.13C NMR
(CDCl.sub.3) .delta. 171.1, 170.5, 170.4, 166.8, 166.7, 165.7,
165.4, 155.2, 154.2, 150.1, 143.2, 143.0, 141.35, 141.3, 137.7,
137.6, 137.2, 137.1, 133.5, 133.4, 129.9, 129.7, 129.5, 128.7,
128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8,
127.7, 127.3, 125.0, 124.8, 120.1, 118.3, 114.6, 98.8, 98.1, 97.8,
97.6, 79.1, 77.9, 75.2, 75.1, 74.99, 74.9, 74.5, 74.4, 73.9, 73.4,
73.2, 72.4, 72.1, 70.3, 69.9, 68.8, 68.3, 67.2, 65.8, 64.9, 63.9,
63.7, 64.9, 63.9, 63.7, 63.1, 62.3, 61.9, 60.4, 55.7, 46.7, 40.6,
40.5, 20.7.
[0367] Synthesis of 29
[0368] Compound 29 is prepared from compound 27 and compound 22
following general procedure E: 8.0 g, 87% (beta), R.sub.f=0.45
(toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.5, 170.4, 169.9, 166.4, 165.1, 164.9, 155.7, 154.1, 150.9,
143.2, 153.0, 141.0, 138.3, 137.3, 137.2, 137.1, 133.8, 135.5,
129.8, 129.5, 129.0, 128.9, 128.8, 128.6, 128.4, 128.35, 128.3,
128.2, 128.0, 127.9, 127.7, 127.7, 127.5, 127.4, 127.2, 125.3,
125.1, 124.9, 120.1, 118.8, 114.5, 101.0, 100.1, 97.9, 97.7, 82.7,
77.9, 77.7, 77.4, 75.4, 75.2, 75.1, 74.7, 74.6, 74.5, 74.3, 74.1,
73.6, 72.4, 70.4, 69.8, 68.8, 64.9, 64.7, 64.2, 62.7, 62.6, 62.0,
61.8, 55.6, 46.7, 40.7, 40.4, 20.75, 20.7
[0369] Synthesis of 30
[0370] Compound 30 is prepared from compound 28 following general
procedure D: 388 mg, 84% yield, TLC R.sub.f=0.45 (EtOAc:petroleum
ether, 1:1). HRMS (ESI) calcd for
C.sub.81H.sub.84Cl.sub.2N.sub.6O.sub.26Na(M+Na).sup.+ m/z 1649.471,
found 1649.4702. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 8.16
(dd, J=6.5 Hz, J=9.0 Hz, 2 H), 8.11 (dd, J=7.1 Hz, J=9.2 Hz, 2 H),
7.52-7.48 (m, 4 H), 7.46-7.39 (m, 4 H), 7.38-7.34 (m, 8 H),
7.33-7.29 (m, 4 H), 7.28-7.23 (m, 4 H), 7.18-7.16 (m, 2 H), 7.05
(d, J=9.2 Hz, 2 H), 6.85 (d, J=9.2 Hz, 2 H), 5.62 (s, 1H), 5.34 (s,
1H), 5.15 (t, J=4.3 Hz, 1H), 5.0 (d, J=11.9 Hz, 1H), 4.86 (d,
J=11.5 Hz, 1H), 4.80 (d, J=11.8 Hz, 1H), 4.77-4.76 (m, 2H), 4.53
(d, J=8.0 Hz, 1H), 4.50 (d, J=7.6 Hz, 1H), 4.47 (d, J=5.7 Hz, 1H),
4.59 (bs, 1H), 4.58-4.54 (m, 1H), 4.53 (d, J=7.6 Hz, 1H), 4.50 (d,
J=8.3 Hz, 1H), 4.47 (d, J=6.0 Hz, 1H), 4.45-4.42 (m, 1H), 4.40-4.39
(m, 1H), 4.35-4.30 (m, 1H), 4.29 (d, J=4.5 Hz, 1H), 4.27-4.24 (m,
1H), 4.23-4.22 (m, 1H), 4.19-4.16 (m, 1H), 4.15 (d, J=2.6 Hz, 1H),
4.12 (d, J=9.8 Hz, 1H), 4.08 (s, 1H), 4.05 (d, J=4.3 Hz, 1H), 4.03
(d, J=4.5 Hz, 1H), 3.97 (d, J=10.3 Hz, 2H), 3.90 (d, J=2.1 Hz, 2H),
3.81 (d, J=3.5 Hz, 2H), 3.76 (s, 3H), 3.74-3.70 (m, 1H), 3.70-3.67
(m, 1H), 3.60 (d, J=10.5 Hz, 1H), 3.56 (d, J=9.7 Hz, 1H), 3.41-3.36
(m, 1H), 3.31 (dd, J=5.7 Hz, J=13.5 Hz, 1H), 3.23 (dd, J=6.8 Hz,
J=13.7 Hz, 1H), 3.06 (d, J=4.5 Hz, 1H), 2.02 (s, 3H), 2.01 (s, 3H);
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.6, 171.1, 170.7,
167.0, 166.8, 165.7, 165.5, 155.2, 150.1, 137.7, 137.6, 137.3,
133.5, 133.4, 129.9, 129.8, 129.5, 128.7, 128.6, 128.5, 128.4,
128.3, 128,1, 127.7, 118.3, 114.6, 98.9, 98.3, 97.8, 97.5, 79.7,
78.9, 76.9, 75.5, 75.2, 74.9, 74.6, 74.4, 73.4, 72.4, 72.1, 71.3,
70.6, 70.3, 68.3, 67.9, 65.7, 64.9, 63.8, 63.6, 63.1, 62,8, 62.4,
60.4, 55.6, 40.5, 20.7.
[0371] Synthesis of 31
[0372] Compound 31 is prepared from compound 29 following general
procedure D: 600 mg, 85%, R.sub.f=0.2 (Toluene/EtOAc 7:3).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.9, 170.5, 166.9,
166.4, 165.1, 164.9, 155.7, 150.9, 138.3, 137.8, 137.3, 137.2,
133.8, 133.5, 129.8, 129.5, 128.9, 128.8, 128.7, 128.6, 128.4,
128.3, 128.2, 128.1, 127.7, 127.5, 127.3, 127.0, 125.3, 118.8,
114.5, 101.0, 100.0, 98.3, 97.7, 82.8, 82.7, 79.2, 77.9, 77.7,
75.4, 75.0, 74.7, 74.6, 74.3, 73.6, 72.5, 71.4, 70.6, 69.8, 64.9,
64.2, 62.7, 62.0, 55.6, 40.4, 20.7
[0373] Synthesis of 32
[0374] Compound 32 is prepared from compound 30 and compound 24
following general procedure E: 1.51 g, 86% yield (beta),
R.sub.f=0.21 (EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.133H.sub.132Cl.sub.3N.sub.9O.sub.40Na(M+Na).sup.+ m/z
2622.7535, found 2622.7549. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.17 (dd, J=6.5 Hz, J=8.9 Hz, 2 H), 8.12-8.08 (m, 4 H),
7.76-7.73 (m, 2H), 7.59 (d, J=7.3 Hz, 2H), 7.54 (d, J=7.7 Hz, 8H),
7.50-7.43 (m, 6H), 7.42-7.35 (m, 12H), 7.34-7.29 (m, 8H), 7.28-7.19
(m, 6H), 7.18-7.13 (m, 4H), 7.08-7.04 (m, 2H), 7.05 (d, J=9.1 Hz,
2H), 6.85 (d, J=8.7 Hz, 2H), 5.63 (d, J=4.6 Hz, 1H), 5.35 (t, J=3.6
Hz, 1H), 5.17-5.12 (m, 1H), 5.10 (d, J=4.6 Hz, 1H), 5.01 (d, J=11.9
Hz, 1H), 4.87 (d, J=7.7 Hz, 1H), 4.84 (d, J=7.7 Hz, 1H), 4.81-4.79
(m, 2H), 4.78-4.75 (m, 3H), 4.74 (d, J=5.9 Hz, 1H), 4.71 (bs, 1H),
4.70 (d, J=3.8 Hz, 1H), 4.63 (d, J=4.0 Hz, 1H), 4.59-4.54 (m, 4H),
4.54-4.49 (m, 4H), 4.48-4.42 (m, 3H), 4.41-4.37 (m, 4H), 4.35-4.29
(m, 4H), 4.28-4.25 (m, 3H), 4.24-4.22 (m, 2H), 4.21-4.17 (m, 2H),
4.16-4.14 (m, 2H), 4.08-4.04 (m, 2H), 4.03-4.01 (m, 1H), 3.97 (d,
J=10.2 Hz, 1H), 3.90 (s, 2H), 3.89 (s, 2H), 3.81 (d, J=4.0 Hz, 2H),
3.75 (s, 3H), 3.74-3.71 (m, 1H), 3.71-3.67 (m, 1H), 3.67-3.64 (m,
1H), 3.64-3.60 (m, 1H), 3.58 (t, J=9.5, 1H), 3.32 (d, J=4.3, 1H),
3.30-3.28 (m, 1H), 3.28 (d, J=3.9, 1H), 2.03 (s, 3H), 2.0 (s, 3H),
2.0 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 171.6,
171.1, 170.5, 170.4, 166.9, 166.8, 166.7, 165.7, 165.5, 165.4,
163.2, 155.2, 154.2, 150.2, 143.2, 143.1, 141.4, 137.7, 137.6,
137.3, 137.2, 137.1, 136.5, 134.1, 133.6, 133.4, 129.9, 129.8,
129.5, 129.4, 128.8, 128.7, 128.6, 128.5, 128.4, 128.35, 128.3,
128.25, 128.2, 128.1, 128.0, 127.9, 127.8, 127.3, 125.0, 124.8,
120.2, 118.3, 114.6, 98.8, 98.3, 98.1, 97.9, 97.8, 97.5, 80.1,
79.0, 78.9, 78.3, 77.9, 76.9, 76.2, 75.8, 75.4, 75.1, 74.9, 74.7,
74.6, 74.5, 74.1, 74.0, 73.8, 73.5, 73.3, 73.2, 72.4, 72.1, 71.4,
70.7, 70.4, 70.3, 70.2, 69.8, 69.5, 68.9, 68.8, 68.3, 67.9, 67.6,
67.1, 65.9, 65.7, 65.2, 64.9, 64.4, 63.9, 63.8, 63.7, 63.6, 63.3,
63.1, 62.3, 62.2, 61.9, 60.4, 55.7, 46.8, 40.7, 40.5, 22.7
[0375] Synthesis of 33
[0376] Compound 33 is prepared from compound 30 and compound 25
following general procedure E: The residue is purified by silica
gel chromatography (EtOAc:petroleum ether, 1:2) to furnish the
disaccharide as a foam, 375 mg, 74% yield, TLC, R.sub.f=0.15
(EtOAc:petroleum ether, 1:2) HRMS (ESI) calcd for
C.sub.125H.sub.128Cl.sub.3N.sub.9O.sub.38Na(M+Na).sup.+ m/z
2490.7324, found 2490.7297. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (dd, J=6.7 Hz, J=9.6 Hz, 2 H), 8.09 (d, J=8.1 Hz, 4
H), 7.50-7.40 (m, 4 H), 7.39-7.34 (m, 6 H), 7.33-7.28 (m, 6H),
7.27-7.20 (m, 10H), 7.20-7.16 (m, 10H), 7.14-7.11 (m, 8H), 7.05 (d,
J=8.9 Hz, 2H), 6.85 (d, J=9.5 Hz, 2H), 5.62 (d, J=4.0 Hz, 1H), 5.35
(d, J=3.7 Hz, 1H), 5.14 (dd, J=5.2 Hz, J=13.1 Hz, 1H), 5.10 (d,
J=3.6 Hz, 1H), 5.07 (d, J=3.7 Hz, 1H), 5.01 (d, J=11.5 Hz, 1H),
4.84 (d, J=4.4 Hz, 2H), 4.82 (t, J=4.3 Hz, 2H), 4.77-4.73 (m, 2H),
4.72 (d, J=3.6 Hz, 2H), 4.71 (d, J=3.7 Hz, 2H), 4.63 (d, J=3.8 Hz,
2H), 4.57-4.53 (m, 2H), 4.53-4.48 (m, 3H), 4.47-4.45 (m, 2H),
4.45-4.43 (m, 2H), 4.42-4.39 (m, 2H), 4.35-4.30 (m, 2H), 4.29 (d,
J=3.7 Hz, 2H), 4.26 (d, J=4.3 Hz, 1H), 4.24-4.18 (m, 1H), 4.18-4.15
(m, 1H), 4.10 (d, J=2.2 Hz, 2H), 4.08-4.05 (m, 2H), 4.04-4.01 (m,
2H), 4.0 (d, J=4.6 Hz, 2H), 3.90 (d, J=10.3 Hz, 2H), 3.92-3.89 (m,
1H), 3.88-3.87 (m, 4H), 3.81 (d, J=4.8 Hz, 2H), 3.77 (s, 3H), 3.73
(d, J=8.8 Hz, 1H), 3.70-3.64 (m, 1H), 3.62 (d, J=10.6 Hz, 1H), 3.58
(d, J=10.6 Hz, 1H), 3.54 (d, J=8.8 Hz, 1H), 3.43 (t, J=9.3 Hz, 1H),
3.30 (d, J=3.9 Hz, 1H), 3.28-3.25 (m, 1H), 3.25-3.23 (m, 1H), 2.013
(s, 3H), 1.99 (s, 3H), 1.97 (s, 3H); .sup.13C NMR (CDCl.sub.3)
.delta. 171.1, 170.5, 170.4, 166.8, 166.7, 165.7, 165.5, 165.4,
164.9, 155.2, 143.2, 143.1, 143.0, 141.3, 137.7, 137.6, 137.3,
137.0, 133.6, 133.4, 129.8, 129.7, 129.5, 129.4, 128.9, 128.8,
128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8,
127.7, 127.6, 127.5, 127.2, 125.0, 124.8, 120.1, 118.2, 114.5,
99.6, 98.8, 98.3, 98.0, 97.9, 97.8, 78.9, 78.8, 77.9, 77.3, 77.0,
76.8, 75.8, 75.3, 75.1, 75.0, 74.9, 74.6, 74.5, 74.4, 74.2, 74.0,
73.8, 73.6, 73.5, 73.4, 73.3, 72.3, 72.2, 72.1, 70.4, 70.3, 70.2,
69.8, 69.1, 68.8, 68.2, 65.7, 65.2, 65.0, 64.9, 64.3, 63.8, 63.7,
63.6, 63.3, 63.1, 62.3, 62.2, 61.9, 60.4, 55.7, 46.8, 40.7, 40.5,
40.3, 20.7.
[0377] Synthesis of 34
[0378] Compound 34 is prepared from compound 31 and compound 22
following general procedure E: 5.25 g, 94% (beta), R.sub.f=0.45
(Toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.4, 169.9, 166.4, 165.9, 165.1, 165.0, 164.9, 155.7, 154.1,
151.0, 143.2, 143.0, 141.3, 138.3, 137.3, 137.2, 137.1, 133.8,
133.5, 129.8, 129.7, 129.5, 129.0, 128.9, 128.6, 128.4, 128.35,
128.3, 128.2, 128.0, 127.9, 127.8, 127.7, 127.6, 127.5, 127.45,
127.4, 127.3, 125.1, 124.9, 120.1, 118.8, 114.5, 100.9, 100.8,
100.1, 97.8, 97.7, 82.8, 82.7, 77.7, 77.6, 77.5, 74.5, 75.3, 75.2,
75.1, 74.9, 74.7, 74.6, 74.3, 74.2, 74.1, 73.6, 72.4, 72.3, 70.4,
69.8, 68.8, 64.9, 64.2, 62.7, 62.6, 61.9, 61.8, 55.6, 46.7, 40.4,
40.3, 20.75, 20.7
[0379] Synthesis of 35
[0380] Compound 35 is prepared from compound 31 and compound 23
following general procedure E: 591 mg, 78% (beta), R.sub.f=0.5
(Toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.5, 166.9, 166.4, 165.1, 165.0, 164.9, 155.7, 150.9, 138.3,
138.2, 137.5, 137.3, 137.3, 137.2, 133.8, 133.7, 133.4, 129.8,
129.7, 129.5, 129.1, 129.0, 128.9, 128.8, 128.6, 128.5, 128.4,
128.3, 128.1, 128.0, 127.7, 127.6, 127.5, 127.4, 125.3, 118.8,
114.5, 100.9, 100.8, 100.0, 98.0, 97.8, 97.7, 82.8, 82.7, 80.1,
77.8, 77.7, 77.5, 75.6, 75.5, 75.2, 75.0, 74.9, 74.7, 74.6, 74.3,
74.2, 74.1, 73.5, 72.4, 72.3, 70.3, 69.8, 65.0, 64.4, 64.2, 63.2,
62.7, 62.6, 62.4, 61.9, 61.8, 55.6, 40.5, 40.4, 40.3, 20.7
[0381] Synthesis of 36
[0382] Compound 36 is prepared from compound 32 following general
procedure D: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:1) to furnish the disaccharide as a foam,
550 mg, 88% yield; TLC, R.sub.f=0.45 (EtOAc:petroleum ether, 1:1).
HRMS (ESI) calcd for
C.sub.118H.sub.122Cl.sub.3N.sub.9O.sub.38Na(M+Na).sup.+ m/z
2400.6854, found 2400.6853. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16-8.07 (m, 6 H), 7.51-7.46 (m, 5 H), 7.46-7.38 (m, 10
H), 7.37-7.22 (m, 8H), 7.21-7.18 (m, 10H), 7.16-7.12 (m, 6H), 7.05
(d, J=10.1 Hz, 2H), 6.84 (d, J=9.3 Hz, 2H), 5.62 (bd, 1H), 5.35 (t,
J=2.1 Hz, 1H), 5.15 (dd, J=3.3 Hz, J=10.0 Hz, 1H), 5.11 (d, J=3.6
Hz, 1H), 5.09 (d, J=3.8 Hz, 1H), 5.0 (d, J=11.5 Hz, 1H), 4.86 (d,
J=7.0 Hz, 1H), 4.83 (d, J=7.5 Hz, 2H), 4.80 (d, J=11.6 Hz, 1H),
4.76-4.73 (m, 2H), 4.72 (d, J=10.4 Hz, 1H), 4.63 (d, J=4.1 Hz, 2H),
4.59-4.54 (m, 3H), 4.53-4.51 (m, 1H), 4.51 (d, J=3.5 Hz, 2H), 4.49
(d, J=10.3 Hz, 2H), 4.45-4.43 (m, 1H), 4.43-4.40 (m, 1H), 4.39-4.38
(m, 2H), 4.37-4.33 (m, 2H), 4.33-4.30 (m, 2H), 4.30-4.27 (m, 2H),
4.27-4.20 (m, 2H), 4.19-4.16 (m, 1H), 4.15 (d, J=2.8 Hz, 1H),
4.12-4.07 (m, 1H), 4.07-4.04 (m, 1H), 4.04 (d, J=4.0 Hz, 1H), 4.02
(d, J=4.5 Hz, 1H), 3.97 (d, J=10.2 Hz, 1H), 3.89-3.87 (m, 4H), 3.81
(d, J=4.2 Hz, 2H), 3.75 (s, 3H), 3.64 (d, J=9.8 Hz, 2H), 3.60 (d,
J=9.1 Hz, 2H), 3.57 (dd, J=4.9 Hz, J=13.3 Hz, 1H), 3.41-3.35 (m,
1H), 3.31 (d, J=3.9 Hz, 1H), 3.29 (t, J=3.25 Hz, 1H), 3.27 (d,
J=3.9 Hz, 1H), 3.23 (d, J=3.9 Hz, 2H), 3.21 (d, J=3.8 Hz, 1H), 3.07
(d, J=4.0 Hz, 1H), 2.02 (s, 3H), 2.016 (s, 3H), 2.01 (s, 3H);
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.6, 171.1, 170.7,
170.6, 167.0, 165.8, 165.7, 165.52, 165.5, 155.2, 150.1, 137.8,
137.6, 137.3, 137.2, 133.5, 133.4, 130.9, 129.9, 129.8, 129.7,
129.5, 129.4, 128.7, 128.66, 128.6, 128.5, 128.4, 128.3, 128.2,
128.1, 128.0, 127.9, 127.7, 127.6, 125.7, 118.3, 114.6, 99.6, 98.9,
98.3, 98.2, 97.8, 97.5, 80.1, 79.8, 78.9, 78.7, 76.9, 75.4, 75.2,
74.9, 74.6, 74.5, 74.3, 74.2, 73.44, 73.4, 72.3, 72.1, 71.7, 71.3,
70.6, 70.5, 70.4, 70.3, 70.2, 68.3, 67.8, 67.6, 65.7, 64.9, 63.9,
63.6, 63.5, 63.3, 63.1, 62.9, 62.3, 62.2, 60.4, 55.7, 40.7, 40.51,
20.7.
[0383] Synthesis of 37
[0384] Compound 37 is prepared from compound 34 following general
procedure D: 2.62 g, 80%, R.sub.f=0.18 (Toluene/EtOAc 7:3).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.9, 170.5, 166.9,
166.4, 165.1, 165.0, 164.9, 155.7, 150.9, 138.3, 138.0, 137.8,
137.3, 137.2, 133.7, 133.5, 129.8, 129.5, 129.0, 128.95, 128.9,
128.8, 128.7, 128.6, 128.4, 128.3, 128.2, 128.1, 127.7, 127.5,
127.45, 127.3, 127.0, 125.3, 118.8, 114.5, 100.9, 100.8, 100.0,
98.3, 97.8, 97.7, 82.8, 82.7, 79.2, 77.7, 77.65, 77.6, 76.55, 75.4,
75.0, 74.9, 74.7, 74.6, 74.3, 73.6, 72.5, 72.3, 71.4, 70.6, 69.8,
64.9, 64.4, 64.2, 62.7, 61.9, 61.8, 55.6, 40.4, 40.3, 20.7
[0385] Synthesis of 38
[0386] Compound 38 is prepared from compound 36 and compound 24
following general procedure E: 272 mg, 80% yield (beta),
R.sub.f=0.15 (EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.170H.sub.170Cl.sub.4N.sub.12O.sub.52Na(M+Na).sup.+ m/z
3373.9672, found 3373.9951. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16 (dd, J=7.0 Hz, J=9.7 Hz, 2 H), 8.12-8.07 (m, 4 H),
7.76 (d, J=7.8 Hz, 1H), 7.60 (d, J=7.6 Hz, 1H), 7.55 (d, J=7.6 Hz,
1H), 7.51 (bd, 1H), 7.49-7.46 (m, 6H), 7.46-7.43 (m, 8 H),
7.43-7.40 (m, 8H), 7.40-7.37 (m, 5H), 7.37-7.35 (m, 4H), 7.35-7.30
(m, 8H), 7.29-7.24 (m, 6H), 7.23-7.19 (m, 4H), 7.19-7.16 (m, 4H),
7.15-7.12 (m, 4H), 7.08-7.05 (m, 2H), 7.05 (d, J=9.1 Hz, 2H), 6.85
(d, J=9.1 Hz, 2H), 5.63 (d, J=3.7 Hz, 1H), 5.35 (t, J=3.6 Hz, 1H),
5.16-5.11 (m, 3H), 5.10 (d, J=4.4 Hz, 1H), 5.08 (d, J=4.0 Hz, 1H),
5.01 (d, J=11.8 Hz, 1H), 4.86-4.79 (m, 8H), 4.78-4.71 (m, 6H), 4.71
(dd, J=4.7 Hz, J=10.2 Hz, 2H), 4.63 (d, J=3.9 Hz, 1H), 4.58-4.53
(m, 6H), 4.53 (d, J=5.5 Hz, 1H), 4.50 (d, J=4.3 Hz, 1H), 4.48-4.46
(m, 4H), 4.45-4.40 (m, 6H), 4.39-4.35 (m, 6H), 4.35-4.31 (m, 4H),
4.30-4.27 (m, 3H), 4.27-4.23 (m, 2H), 4.23-4.19 (m, 2H), 4.19-4.15
(m, 2H), 4.10-4.07 (m, 2H), 4.06-4.0 (m, 1H), 3.97 (d, J=11.4 Hz,
1H), 3.90-3.87 (m, 4H), 3.81 (d, J=4.7 Hz, 2H), 3.76 (s, 3H),
3.75-3.74 (m, 1H), 3.73-3.65 (m, 1H), 3.64 (q, J=8.1 Hz, 2H), 3.57
(t, J=8.8, 1H), 3.32 (d, J=3.5, 1H), 3.31-3.27 (m, 1H), 3.27 (t,
J=3.0, 1H), 2.03 (s, 3H), 2.01 (s, 3H), 2.0 (s, 3H), 1.99 (3H);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.5, 170.4,
166.9, 166.8, 166.77, 166.7, 165.6, 165.5, 165.4, 155.3, 154.2,
150.2, 143.2, 143.1, 141.4, 137.7, 137.6, 137.3, 137.2, 137.16,
137.1, 133.6, 133.5, 129.9, 129.8, 129.5, 129.4, 128.8, 128.7,
128.5, 128.4, 128.35, 128.3, 128.27, 128.2, 128.1, 128.0, 127.9,
127.8, 127.7, 127.3, 125.1, 124.9, 120.2, 118.3, 114.6, 98.8, 98.3,
98.2, 98.1, 98.06, 98.0, 97.9, 97.8, 97.6, 79.0, 78.9, 78.7, 77.9,
76.9, 75.4, 75.3, 75.1, 75.0, 74.9, 74.7, 74.6, 74.3, 74.2, 74.0,
73.9, 73.5, 73.4, 72.4, 72.1, 70.4, 70.3, 70.2, 69.8, 68.3, 67.7,
67.5, 67.2, 65.7, 64.9, 64.7, 63.9, 63.8, 63.77, 63.7, 63.6, 63.58,
63.5, 63.1, 62.3, 62.2, 61.9, 55.7, 46.8, 40.7, 40.5, 20.8.
[0387] Synthesis of 39
[0388] Compound 39 is prepared from compound 36 and compound 25
following general procedure E: The residue is purified by silica
gel chromatography (EtOAc:petroleum ether, 1:2) to furnish the
disaccharide as a foam, 260 mg, 82% yield, R.sub.f=0.22
(EtOAc:petroleum ether, 2:3). HRMS (ESI) calcd for
C.sub.162H.sub.170Cl.sub.4N.sub.12O.sub.50Na(M+Na).sup.+ m/z
3245.9781, found 3245.9404. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (dd, J=6.8 Hz, J=9.5 Hz, 2 H), 8.10-8.06 (m, 4 H),
7.50-7.46 (m, 4 H), 7.45-7.40 (m, 8 H), 7.40-7.34 (m, 8 H),
7.33-7.28 (m, 12H), 7.27-7.22 (m, 10H), 7.21-7.16 (m, 8H),
7.14-7.11 (m, 9H), 7.05 (d, J=9.5 Hz, 2H), 6.85 (d, J=8.9 Hz, 2H),
5.62 (d, J=3.4 Hz, 1H), 5.34 (t, J=3.7 Hz, 2H), 5.14-5.10 (m, 2H),
5.10 (t, J=3.9 Hz, 2H), 5.07 (d, J=4.1 Hz, 1H), 5.01 (d, J=11.6 Hz,
1H), 4.84-4.81 (m, 4H), 4.81-4.78 (m, 2H), 4.76-4.73 (m, 2H),
4.73-4.70 (m, 2H), 4.70 (d, J=5.0 Hz, 1H), 4.67 (bs, 2H), 4.63 (d,
J=3.8 Hz, 1H), 4.57-4.53 (m, 2H), 4.53-4.48 (m, 3H), 4.48 (d, J=3.5
Hz, 1H), 4.46 (d, J=2.5 Hz, 1H), 4.44 (d, J=3.0 Hz, 2H), 4.42-4.40
(m, 4H), 4.40-4.38 (m, 4H), 4.35-4.29 (m, 2H), 4.29-4.24 (m, 4H),
4.24-4.18 (m, 4H), 4.18-4.15 (m, 2H), 4.08-4.05 (m, 2H), 4.05-3.99
(m, 4H), 3.97 (d, J=10.4 Hz, 1H), 3.88-3.87 (m, 6H), 3.81 (d, J=4.8
Hz, 2H), 3.77 (s, 3H), 3.75 (d, J=3.7 Hz, 1H), 3.73 (d, J=3.5 Hz,
1H), 3.71 (d, J=3.5 Hz, 1H), 3.69-3.64 (m, 2H), 3.62 (d, J=3.1 Hz,
1H), 3.60-3.58 (m, 2H), 3.58 (d, J=3.5 Hz, 1H), 3.56 (t, J=9.7 Hz,
2H), 3.44 (t, J=9.7 Hz, 2H), 3.30 (d, J=3.7 Hz, 1H), 3.28-3.23 (m,
1H), 2.01 (s, 6H), 1.99 (s, 3H), 1.97 (s, 3H); .sup.13C NMR
(CDCl.sub.3) .delta. 170.5, 166.9, 166.8, 165.7, 165.5, 165.4,
155.2, 150.1, 137.7, 137.6, 137.4, 137.2, 137.1, 133.4, 129.8,
129.7, 129.5, 129.4, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1,
128.0, 127.9, 127.7, 127.6, 127.5, 118.3, 114.5, 98.7, 98.3, 98.1,
98.0, 97.9, 97.8, 97.7, 97.5, 80.4, 78.9, 78.7, 78.6, 77.7, 75.4,
75.3, 75.2, 75.1, 74.9, 74.6, 74.5, 74.3, 74.2, 74.1, 74.0, 73.5,
72.4, 72.1, 70.3, 70.2, 70.1, 68.3, 67.7, 67.5, 65.7, 64.9, 63.8,
63.7, 63.5, 62.6, 62.1, 55.7, 40.6, 40.5, 20.7.
[0389] Synthesis of 40
[0390] Compound 40 is prepared from compound 37 and compound 22
following general procedure E: 1.4 g, quant (beta), R.sub.f=0.4
(Toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.6, 169.8, 166.6, 166.0, 165.9, 165.4, 165.1, 155.6, 154.1,
151.0, 143.2, 142.9, 141.3, 141.2, 137.9, 137.7, 137.3, 137.2,
137.0, 133.4, 133.2, 129.8, 129.7, 129.6, 129.5, 129.4, 129.0,
128.6, 128.4, 128.3, 128.2, 128.15, 128.1, 128.0, 127.9, 127.8,
127.7, 127.6, 125.3, 124.9, 120.0, 118.9, 114.5, 100.3, 98.3, 97.8,
97.7, 97.6, 97.5, 97.1, 82.8, 78.6, 78.5, 75.3, 75.1, 74.9, 74.1,
74.0, 73.8, 73.6, 73.4, 73.2, 72.7, 71.5, 70.5, 70.3, 70.1, 69.5,
68.6, 68.0, 67.4, 63.5, 63.4, 62.9, 62.8, 62.5, 62.2, 62.0, 61.9,
59.3, 59.2, 55.6, 46.7, 40.7, 20.7
[0391] Synthesis of 41
[0392] Compound 34 is prepared from compound 37 and compound 23
following general procedure E: 785 mg, 88% (beta), R.sub.f=0.48
(toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.4, 166.9, 166.4, 165.1, 165.0, 164.9, 155.7, 150.9, 138.2,
137.5, 137.2, 133.8, 133.7, 133.4, 129.8, 129.7, 129.5, 129.0,
128.9, 128.8, 128.6, 128.5, 128.3, 128.2, 128.1, 128.0, 127.7,
127.6, 127.5, 118.8, 114.5, 100.9, 100.8, 100.7, 100.0, 98.0, 97.8,
97.8, 97.7, 82.8, 82.7, 80.1, 77.8, 77.7, 77.6, 75.6, 75.5, 75.4,
75.2, 75.0, 74.9, 74.7, 74.6, 74.3, 74.2, 73.5, 72.4, 72.3, 70.3,
69.8, 65.0, 64.4, 64.2, 63.2, 62.7, 62.7, 62.6, 62.4, 61.9, 61.8,
61.7, 55.6, 40.5, 40.4, 40.3, 40.3, 20.7
[0393] Synthesis of 42
[0394] Compound 42 is prepared from compound 38 following general
procedure D: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:1) to furnish the disaccharide as a foam,
170 mg, 86% yield, R.sub.f=0.45 (EtOAc:petroleum ether, 1:1). HRMS
(ESI) calcd for
C.sub.155H.sub.160Cl.sub.4N.sub.12O.sub.50Na(M+Na).sup.+ m/z
3151.8998, found 3151.8971. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16 (dd, J=6.8 Hz, J=9.4 Hz, 2H), 8.11-8.07 (m, 6H),
7.51-7.38 (m, 6 H), 7.37-7.27 (m, 5 H), 7.27-7.16 (m, 5 H),
7.46-7.38 (m, 8 H), 7.37-7.22 (m, 10H), 7.21-7.18 (m, 12H), 7.15
(dd, J=6.1 Hz, J=8.4 Hz, 6H), 7.05 (d, J=8.9 Hz, 2H), 6.85 (d,
J=9.1 Hz, 2H), 5.63 (bd, 1H), 5.35 (t, J=3.4 Hz, 1H), 5.15-5.11 (m,
2H), 5.11-5.06 (m, 2H), 5.01 (d, J=11.4 Hz, 1H), 4.85-4.79 (m, 2H),
4.78-4.74 (m, 5H), 4.74-4.66 (m, 10H), 4.63 (d, J=4.1 Hz, 2H),
4.58-4.54 (m, 8H), 4.53 (d, J=6.8 Hz, 2H), 4.51 (d, J=6.6 Hz, 4H),
4.47-4.43 (m, 6H), 4.43-4.39 (m, 4H), 4.36-4.26 (m, 4H), 4.26-4.19
(m, 2H), 4.19-4.16 (m, 2H), 4.09-4.06 (m, 1H), 4.05-3.99 (m, 2H),
3.97 (d, J=10.4 Hz, 1H), 3.89-3.86 (m, 4H), 3.82 (d, J=4.8 Hz, 2H),
3.76 (s, 3H), 3.75-3.71 (m, 2H), 3.71-3.65 (m, 2H), 3.63-3.57 (m,
2H), 3.56 (t, J=10.1 Hz, 1H), 3.44 (t, J=9.7 Hz, 1H), 3.31 (d,
J=4.1 Hz, 1H), 3.29-3.24 (m, 2H), 2.01 (s, 3H), 2.01 (s, 3H), 2.0
(s, 3H), 2.0 (s, 3H) .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
169.9, 168.9, 168.74, 168.7, 165.2, 164.9, 163.9, 163.7, 163.6,
153.4, 148.3, 135.9, 135.8, 135.4, 135.37, 135.35, 131.7, 128.0,
127.9, 127.7, 127.6, 126.8, 126.7, 126.6, 126.57, 126.5, 126.3,
126.28, 126.2, 125.9, 125.87, 125.8, 116.4, 112.7, 97.1, 96.4,
96.3, 96.1, 95.9, 95.7, 77.9, 77.2, 76.9, 73.6, 73.5, 73.47, 73.4,
73.3, 73.1, 72.8, 72.5, 72.3, 72.2, 71.6, 70.6, 70.3, 69.5, 68.7,
68.5, 68.4, 66.5, 65.9, 65.8, 65.7, 63.9, 63.1, 62.0, 61.8, 61.7,
61.6, 61.3, 60.9, 60.5, 60.3, 53.9, 38.8, 38.7, 18.9.
[0395] Synthesis of 136
[0396] Compound 136 is prepared from compound 137 following general
procedure D: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 2:3) to furnish the disaccharide as a foam,
890 mg, 229 .mu.mol, 97% yield; TLC (EtOAc:petroleum ether, 1:1,
v/v): R.sub.f=0.45; HRMS (ESI) calcd for
C.sub.192H.sub.198Cl.sub.5N.sub.15O.sub.62 Na.sub.2(M+2Na).sup.2+
m/z 1965.054, found 1965.0496. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16-8.13 (m, 2H), 8.12-8.07 (m, 6H), 7.52-7.47 (m, 6 H),
7.45-7.39 (m, 12 H), 7.38-7.34 (m, 15 H), 7.32-7.27 (m, 10 H),
7.27-7.22 (m, 8 H), 7.21-7.16 (m, 10H), 7.14-7.11 (m, 6H), 7.05 (d,
J=8.9 Hz, 2H), 6.85 (d, J=9.1 Hz, 2H), 5.62 (d, J=3.5 Hz, 1H), 5.35
(d, J=3.4 Hz, 1H), 5.15-5.11 (m, 2H), 5.10-5.06 (m, 2H), 5.01 (d,
J=11.4 Hz, 1H), 4.86-4.79 (m, 2H), 4.77-4.71 (m, 6H), 4.70-4.66 (m,
10H), 4.63 (d, J=4.0 Hz, 1H), 4.58-4.53 (m, 6H), 4.53-4.51 (m, 2H),
4.50 (d, J=3.5 Hz, 1H), 4.49-4.47 (m, 6H), 4.46-4.39 (m, 8H),
4.36-4.33 (m, 4H), 4.32-4.27 (m, 6H), 4.26-4.21 (m, 4H), 4.20-4.15
(m, 3H), 4.13-4.12 (m, 2H), 4.11 (d, J=7.1 Hz, 1H), 4.09-4.07 (m,
1H), 4.06-3.99 (m, 2H), 3.97-3.95 (m, 1H), 3.94-3.93 (m, 1H),
3.88-3.87 (m, 4H), 3.87-3.86 (m, 4H), 3.81 (d, J=5.0 Hz, 2H), 3.76
(s, 3H), 3.73-3.65 (m, 4H), 3.62-3.57 (m, 2H), 3.56-3.52 (m, 2H),
3.40-3.35 (m, 2H), 3.31 (d, J=4.1 Hz, 1H), 3.29-3.26 (m, 2H), 3.26
(d, J=4.3 Hz, 1H), 3.23 (d, J=3.7 Hz, 1H), 3.21 (d, J=3.7 Hz, 1H),
2.94 (d, J=4.8 Hz, 1H), 2.03 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H),
2.0 (s, 3H), 1.99 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 171.6, 171.1, 170.7, 170.6, 170.5, 167.0, 166.8, 165.7,
165.5, 155.2, 150.2, 137.7, 137.6, 137.3, 137.2, 133.5, 129.9,
129.7, 129.5, 129.4, 128.7, 128.6, 128.5, 128.4, 128.3, 128.1,
127.7, 127.6, 118.3, 114.6, 98.9, 98.3, 98.1, 97.9, 97.8, 97.7,
97.5, 79.7, 78.9, 78.7, 75.4, 75.3, 75.2, 75.1, 74.9, 74.6, 74.3,
74.1, 73.5, 72.4, 72.1, 71.3, 70.6, 70.5, 70.4, 70.3, 70.2, 68.3,
67.8, 67.6, 67.5, 65.7, 64.9, 63.8, 63.6, 63.5, 63.1, 62.8, 62.3,
62.1, 60.4, 55.7, 40.6, 40.5, 21.0, 20.7
[0397] Synthesis of 43
[0398] Compound 43 is prepared from compound 40 following general
procedure D: 960 mg, 72%, R.sub.f=0.15 (toluene/EtOAc 7:3);
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 171.9, 170.5, 170.4,
166.9, 166.4, 165.1, 165.0, 164.9, 155.7, 150.9, 138.3, 138.2,
138.0, 137.8, 137.3, 137.2, 133.7, 133.5, 129.8, 129.5, 129.0,
128.95, 128.9, 128.8, 128.7, 128.6, 128.4, 128.3, 128.2, 128.1,
127.7, 127.5, 127.45, 127.3, 127.0, 125.3, 118.8, 114.5, 100.9,
100.8, 100.7, 100.0, 98.3, 97.8, 97.7, 82.9, 82.8, 82.7, 79.2,
77.7, 77.65, 77.6, 76.55, 75.4, 75.0, 74.9, 74.7, 74.6, 74.3, 73.6,
72.5, 72.3, 71.4, 70.6, 69.8, 64.9, 64.4, 64.2, 62.7, 62.6, 61.9,
61.8, 61.7, 55.6, 40.4, 40.3, 20.7
[0399] Synthesis of 44
[0400] Compound 44 is prepared from compound 42 and compound 25
following general procedure E: The residue is purified by silica
gel chromatography (EtOAc:petroleum ether, 1:2) to furnish the
disaccharide as a foam, 173 mg, 89% yield, R.sub.f=0.23
(EtOAc:petroleum ether, 1:2). HRMS (ESI) calcd for
C.sub.199H.sub.204Cl.sub.5N.sub.15O.sub.62Na(M+Na).sup.+ m/z
3993.16, found 3993.1611. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.
8.16 (dd, J=7.0 Hz, J=9.7 Hz, 2 H), 8.11-8.07 (m, 6 H), 7.51-7.47
(m, 8 H), 7.45-7.38 (m, 10 H), 7.37-7.33 (m, 14 H), 7.33-7.27 (m,
12H), 7.26-7.19 (m, 10H), 7.18-7.16 (m, 10H), 7.14-7.12 (m, 8H),
7.05 (d, J=9.1 Hz, 2H), 6.85 (d, J=9.1 Hz, 2H), 5.63 (d, J=4.0 Hz,
1H), 5.35 (t, J=3.8 Hz, 2H), 5.15-5.11 (m, 2H), 5.10-5.06 (m, 2H),
5.01 (d, J=11.4 Hz, 1H), 4.85-4.82 (m, 4H), 4.81-4.79 (m, 2H),
4.78-4.74 (m, 2H), 4.73-4.71 (m, 4H), 4.71-4.69 (m, 1H), 4.68-4.66
(m, 10H), 4.63 (d, J=3.9 Hz, 1H), 4.58-4.53 (m, 10H), 4.53-4.51 (m,
6H), 4.51 (d, J=3.9 Hz, 1H), 4.49-4.47 (m, 2H), 4.46-4.43 (m, 6H),
4.43-4.41 (m, 6H), 4.41-4.39 (m, 2H), 4.35-4.26 (m, 2H), 4.25-4.20
(m, 4H), 4.19-4.16 (m, 4H), 4.09-3.99 (m, 2H), 3.97 (d, J=9.9 Hz,
1H), 3.89-3.86 (m, 8H), 3.82 (d, J=5.4 Hz, 2H), 3.76 (s, 3H), 3.75
(bd, 1H), 3.74 (bd, 1H), 3.72 (bd, 1H), 3.70-3.65 (m, 2H),
3.63-3.58 (m, 2H), 3.58 (d, J=3.6 Hz, 1H), 3.56 (t, J=9.2 Hz, 1H),
3.44 (t, J=9.6 Hz, 2H), 3.31 (d, J=4.1 Hz, 1H), 3.29-3.27 (m, 1H),
3.27-3.24 (m, 1H), 2.014 (s, 3H), 2.012 (s, 3H), 2.0 (s, 3H), 1.99
(s, 3H), 1.97 (s, 3H); .sup.13C NMR (CDCl.sub.3) .delta. 168.8,
168.7, 168.6, 168.5, 165.1, 165.0, 164.9, 163.9, 163.7, 163.6,
163.5, 153.4, 148.4, 135.9, 135.8, 135.7, 135.5, 135.4, 135.3,
131.7, 131.6, 128.3, 128.1, 127.9, 127.7, 126.9, 126.8, 126.7,
126.6, 126.5, 126.4, 126.3, 126.2, 126.1, 125.9, 125.8, 116.5,
112.8, 96.9, 96.5, 96.3, 96.2, 96.1, 95.9, 95.8, 95.7, 78.6, 77.2,
77.0, 76.9, 75.9, 75.6, 75.3, 73.6, 73.5, 73.4, 73.3, 73.1, 72.8,
72.7, 72.5, 72.3, 71.7, 71.6, 70.6, 70.3, 68.6, 68.5, 68.4, 68.3,
66.5, 65.9, 65.8, 65.7, 65.6, 63.9, 63.1, 62.1, 61.9, 61.8, 61.7,
60.8, 60.5, 60.3, 53.9, 38.9, 38.7, 20.9.
[0401] Synthesis of 137
[0402] Compound 137 is prepared from compound 24 and compound 42
following general procedure E: 1.16 g, 0.282 mmol, 86% yield; TLC,
(EtOAc:petroleum ether, 1:2 v/v): R.sub.f=0.2. HRMS (ESI) calcd for
C.sub.207H.sub.208Cl.sub.5N.sub.15O.sub.64Na(M+Na).sup.+ m/z
4129.1865, found 4129.1802. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.17 (dd, J=6.7 Hz, J=9.6 Hz, 2 H), 8.12-8.08 (m, 4 H),
7.75 (d, J=2.5 Hz, 2H), 7.73 (d, J=2.5 Hz, 2H), 7.59 (d, J=7.4 Hz,
2H), 7.54 (d, J=7.6 Hz, 2H), 7.50-7.45 (m, 10H), 7.44-7.38 (m, 14
H), 7.37-7.29 (m, 12H), 7.28-7.21 (m, 10H), 7.20-7.17 (m, 8H),
7.16-7.13 (m, 4H), 7.08-7.06 (m, 3H), 7.06 (d, J=9.0 Hz, 2H), 6.85
(d, J=9.0 Hz, 2H), 5.63 (d, J=3.7 Hz, 1H), 5.35 (t, J=3.6 Hz, 2H),
5.17-5.07 (m, 5H), 5.01 (d, J=12.0 Hz, 1H), 4.88-4.81 (m, 8H),
4.78-4.68 (m, 10H), 4.59-4.53 (m, 6H), 4.53 (d, J=5.0 Hz, 1H), 4.51
(d, J=5.0 Hz, 1H), 4.49-4.41 (m, 8H), 4.41-4.37 (m, 12H), 4.36-4.28
(m, 8H), 4.28-4.22 (m, 6H), 4.21-4.18 (m, 4H), 4.17-4.13 (m, 4H),
4.11-4.0 (m, 3H), 3.98 (d, J=10.5 Hz, 1H), 3.91-3.87 (m, 6H), 3.81
(d, J=5.0 Hz, 2H), 3.73 (s, 3H), 3.71-3.65 (m, 2H), 3.64-3.58 (m,
2H), 3.58-3.52 (m, 2H), 3.32-3.26 (m, 3H), 2.03 (s, 3H), 2.01 (s,
6H), 2.0 (s, 3H), 1.99 (3H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.6, 170.5, 166.8, 165.7, 165.6, 165.5, 155.2, 154.2,
150.2, 143.2, 143.1, 141.4, 137.7, 137.6, 137.3, 137.2, 137.1,
133.5, 129.9, 129.8, 129.6, 129.5, 128.8, 128.7, 128.5, 128.4,
128.3, 128.2, 128.0, 127.9, 127.8, 127.3, 125.1, 124.8, 120.2,
118.3, 114.6, 98.8, 98.3, 98.2, 98.1, 97.9, 97.8, 97.5, 79.1, 78.9,
78.8, 75.4, 75.3, 75.1, 75.0, 74.7, 74.6, 74.2, 74.0, 73.9, 73.5,
73.4, 72.4, 72.1, 70.4, 70.3, 69.8, 68.8, 67.6, 67.2, 66.4, 65.7,
64.9, 63.9, 63.7, 63.6, 63.2, 62.4, 62.2, 62.0, 60.4, 55.7, 40.7,
40.6, 20.8, 20.7.
[0403] Synthesis of 138
[0404] Compound 138 is prepared from compound 25 and compound 136
following general procedure E: The residue is purified by silica
gel chromatography (EtOAc:petroleum ether, 1:2) to furnish the
disaccharide as a foam, 780 mg, 0.165 mmol, 92% yield; TLC
(EtOAc:petroleum ether, 1:2, v/v): R.sub.f=0.18; HRMS (ESI) calcd
for
C.sub.236H.sub.242Cl.sub.6N.sub.18O.sub.74Na.sub.2(M+2Na).sup.2+
m/z 2386.1851, found 2386.1833. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16 (dd, J=7.0 Hz, J=9.6 Hz, 2 H), 8.12-8.07 (m, 6 H),
7.51-7.46 (m, 8 H), 7.45-7.37 (m, 12 H), 7.37-7.34 (m, 18 H),
7.34-7.29 (m, 22H), 7.29-7.27 (m, 12H), 7.26-7.20 (m, 10H),
7.20-7.16 (m, 10H), 7.15-7.12 (m, 5H), 7.06 (d, J=9.2 Hz, 2H), 6.85
(d, J=9.2 Hz, 2H), 5.63 (d, J=4.0 Hz, 1H), 5.35 (t, J=3.9 Hz, 2H),
5.17-5.06 (m, 6H), 5.0 (d, J=12.0 Hz, 1H), 4.86-4.79 (m, 12H),
4.81-4.79 (m, 10H), 4.78-4.66 (m, 14H), 4.65 (d, J=4.6 Hz, 1H),
4.59-4.54 (m, 6H), 5.54 (t, J=10.2 Hz, 2H), 4.48-4.40 (m, 10H),
4.37-4.27 (m, 8H), 4.27-4.20 (m, 6H), 4.20-4.17 (m, 4H), 4.10-4.0
(m, 4H), 3.97-3.95 (m, 1H), 3.95-3.93 (m, 2H), 3.93-3.91 (m, 3H),
3.90-3.87 (m, 4H), 3.86-3.84 (m, 6H), 3.82 (d, J=4.2 Hz, 6H), 3.74
(s, 3H), 3.73-3.71 (m, 1H), 3.70-3.67 (m, 2H), 3.64-3.58 (m, 3H),
3.57-3.52 (m, 2H), 3.45-3.40 (m, 2H), 3.33-3.25 (m, 3H), 2.01 (s,
6H), 2.0 (s, 6H), 1.99 (s, 3H), 1.97 (s, 3H); .sup.13C-NMR (125
MHz, CDCl.sub.3) .delta. 170.7, 170.6, 170.5, 167.0, 166.9, 166.8,
165.8, 165.7, 165.6, 155.4, 150.3, 137.8, 137.7, 137.6, 137.4,
137.3, 133.6, 133.5, 129.9, 129.8, 129.6, 128.8, 128.7, 128.6,
128.5, 128.4, 128.36, 128.3, 128.2, 128.1, 128.0, 127.8, 127.7,
118.4, 116.6, 114.7, 98.9, 98.4, 98.3, 98.2, 98.1, 98.0, 97.9,
97.7, 91.9, 80.5, 79.1, 78.9, 78.8, 77.4, 77.1, 76.9, 75.5, 75.4,
75.3, 75.2, 75.1, 75.0, 74.7, 74.6, 74.3, 74.2, 73.6, 73.5, 72.5,
72.3, 70.5, 70.4, 70.2, 68.6, 68.4, 67.7, 67.6, 66.6, 66.5, 66.4,
66.3, 65.0, 63.9, 63.8, 63.7, 63.6, 62.7, 62.4, 62.3, 55.7, 40.8,
40.7, 19.3.
[0405] Synthesis of 45
[0406] Compound 45 is prepared from compound 43 and compound 23
following general procedure E: 1.05 g, 89% (beta), R.sub.f=0.46
(Toluene/EtOAc 4:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.45, 170.44, 170.43, 170.38, 166.9, 166.4, 166.4, 165.1, 165.0,
164.9, 155.7, 150.9, 138.3, 138.26, 138.25 137.9, 137.5, 137.35,
137.3, 137.28, 137.26, 133.8, 133.7, 133.5, 129.8, 129.7, 129.5,
129.1, 129.0, 128.9, 128.8, 128.62, 128.59, 128.57, 128.4, 128.3,
128.2, 128.1, 128.0, 127.7, 127.6, 127.5, 127.4, 118.8, 114.5,
100.9, 100.8, 100.7 (2C), 100.0, 98.1, 97.9, 97.8 (2C), 97.7, 82.8,
82.7, 80.1, 77.8, 77.7, 77.6, 77.6, 77.5, 75.6, 75.5, 75.4, 75.2,
75.2, 75.1, 74.9, 74.7, 74.6, 74.3, 74.2, 74.2, 73.6, 72.4, 72.3,
70.4, 69.8, 65.0, 64.4, 64.2, 63.2, 62.7, 62.7, 62.6, 62.4, 61.9,
61.8, 61.7, 55.6, 40.5, 40.4, 40.3, 40.3, 20.8, 20.7
##STR00037## ##STR00038##
[0407] General Procedure F (GPF): Selective De-chloroacetylation.
DABCO (6 equiv. per chloroacetyl group) is added to the starting
material in dry ethanol (5 mL for 40 .mu.mol) at room temperature.
The mixture is heated at 60-70.degree. C. under argon for 2 h.
After TLC (EtOAc:petroleum ether, 3:2) indicated the completion of
the reaction Dowex 50WX8-200 ion-exchange resin is added to
neutralize the solution. After 15 min the resin is filtered off and
the solution is concentrated to dryness. Chromatography
(EtOAc:petroleum ether, 3:2) affords the products with unblocked
primary hydroxyl groups.
[0408] Synthesis of 46
[0409] Compound 46 is prepared from compound 33 following general
procedure F: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:1) to furnish the disaccharide as a foam,
330 mg, 98% yield, R.sub.f=0.2 (EtOAc:petroleum ether, 1:1). HRMS
(ESI) calcd for C.sub.119H.sub.125N.sub.9O.sub.35Na (M+Na).sup.+
m/z 2262.8176, found 2262.8201. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (dd, J=6.2 Hz, J=9.6 Hz, 2 H), 8.10-8.07 (m, 4 H),
7.47-7.44 (m, 4 H), 7.43-7.41 (m, 6 H), 7.40-7.38 (m, 4H),
7.37-7.32 (m, 4 H), 7.31-7.25 (m, 6H), 7.24-7.21 (m, 8H), 7.20-7.17
(m, 6H), 7.16-7.14 (m, 6H), 7.06 (d, J=9.5 Hz, 2H), 6.85 (d, J=8.9
Hz, 2H), 5.64 (d, J=4.0 Hz, 1H), 5.37 (t, J=3.1 Hz, 2H), 5.15 (t,
J=3.3 Hz, 2H), 5.12 (t, J=2.6 Hz, 2H), 5.06 (d, J=2.6 Hz, 1H), 5.02
(d, J=6.6 Hz, 1H), 4.98 (bs, 1H), 4.88 (d, J=5.7 Hz, 1H), 4.85 (d,
J=6.2 Hz, 1H), 4.82 (d, J=11.9 Hz, 1H), 4.76 (d, J=2.7 Hz, 1H),
4.74 (d, J=2.7 Hz, 1H), 4.72 (t, J=3.0 Hz, 2H), 4.68 (d, J=3.7 Hz,
1H), 4.63 (d, J=4.0 Hz, 1H), 4.51 (d, J=5.5 Hz, 1H), 4.49-4.46 (m,
2H), 4.46-4.44 (m, 2H), 4.39 (d, J=10.2 Hz, 1H), 4.33 (d, J=2.7 Hz,
1H), 4.31 (d, J=2.7 Hz, 1H), 4.29-4.25 (m, 2H), 4.25-4.21 (m, 2H),
4.25-4.17 (m, 2H), 4.16 (d, J=4.7 Hz, 1H), 4.14-4.10 (m, 1H), 4.05
(q, J=3.7 Hz, 2H), 3.98 (d, J=10.5 Hz, 1H), 3.96-3.93 (m, 1H),
3.87-3.82 (m, 1H), 3.79-3.76 (m, 3H), 3.75 (s, 3H), 3.73 (d, J=2.4
Hz, 1H), 3.68 (d, J=4.0 Hz, 1H), 3.66 (d, J=9.2 Hz, 1H), 3.62-3.57
(m, 1H), 3.57 (d, J=3.6 Hz, 1H), 3.53 (q, J=7.6 Hz, 2H), 3.42 (t,
J=9.3 Hz, 2H), 3.35 (d, J=3.8 Hz, 1H), 3.33 (d, J=3.8 Hz, 1H), 3.32
(d, J=3.8 Hz, 1H), 3.30 (d, J=3.8 Hz, 1H), 3.28 (d, J=3.8 Hz, 1H),
3.27 (d, J=3.6 Hz, 1H), 3.25-3.22 (m, 1H), 2.01 (s, 3H), 2.00 (s,
3H), 1.98 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.6, 170.5, 165.8, 165.7, 155.2, 150.4, 137.7, 137.6, 137.5,
137.4, 133.4, 133.3, 129.9, 129.8, 129.7, 129.6, 128.7, 128.6,
128.5, 128.4, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8, 127.7,
127.5, 118.3, 114.7, 98.2, 97.9, 97.8, 97.7, 97.3, 97.2, 80.6,
79.3, 79.2, 77.8, 77.6, 76.8, 75.2, 75.1, 75.0, 74.6, 74.2, 74.0,
73.9, 73.4, 73.3, 73.2, 73.0, 72.9, 72.9, 72.3, 72.1, 70.5, 70.1,
70.0, 69.5, 69.2, 68.9, 68.1, 67.9, 67.8, 64.1, 63.9, 63.8, 63.1,
62.8, 62.7, 62.4, 61.6, 61.2, 61.1, 55.7, 20.8, 20.7.
[0410] Synthesis of 47
[0411] Compound 47 is prepared from compound 35 following general
procedure F: 342 mg, 74%, R.sub.f=0.17 (Toluene/EtOAc 7:3).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.4, 165.0,
155.6, 151.1, 138.2, 138.1, 137.8, 137.6, 137.4, 137.3, 133.6,
133.5, 133.3, 129.8, 129.6, 129.1, 129.0, 128.7, 128.6, 128.5,
128.4, 128.3, 128.2, 128.1, 128.0, 128.0, 127.9, 127.7, 127.6,
127.5, 127.2, 127.1, 118.4, 114.6, 101.1, 101.0, 100.4, 97.6, 97.3,
97.2, 83.5, 83.35, 83.3, 80.3, 78.0, 77.9, 77.8, 77.7, 77.6, 75.5,
75.2, 75.1, 75.0, 74.9, 74.8, 74.5, 74.4, 73.9, 73.4, 73.3, 72.3,
70.0, 69.6, 69.5, 63.2, 62.9, 62.7, 62.6, 62.2, 62.1, 61.8, 61.5,
61.4, 55.6, 20.8
[0412] Synthesis of 48
[0413] Compound 48 is prepared from compound 39 following general
procedure F: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 3:2) to furnish the disaccharide as a foam,
99 mg, 84% yield, R.sub.f=0.25 (EtOAc:petroleum ether, 3:2). HRMS
(ESI) calcd for C.sub.154H.sub.162N.sub.12O.sub.46Na (M+Na).sup.+
m/z 2938.0604, found 2938.0603. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (dd, J=6.1 Hz, J=9.7 Hz, 2 H), 8.10-8.07 (m, 4 H),
7.46-7.44 (m, 4 H), 7.43-7.39 (m, 6 H), 7.39-7.36 (m, 4H),
7.36-7.33 (m, 4 H), 7.33-7.30 (m, 6H), 7.30-7.26 (m, 8H), 7.24-7.21
(m, 2H), 7.20-7.17 (m, 6H), 7.16-7.14 (m, 6H), 7.07 (d, J=9.5 Hz,
2H), 6.84 (d, J=9.1 Hz, 2H), 5.64 (d, J=4.0 Hz, 1H), 5.36 (t, J=3.1
Hz, 2H), 5.15 (t, J=2.6 Hz, 2H), 5.13-5.10 (m, 2H), 5.07 (dd, J=4.2
Hz, J=7.8 Hz, 2H), 5.02 (d, J=12.5 Hz, 1H), 4.88 (d, J=7.2 Hz, 1H),
4.86 (bd, 1H), 4.85 (d, J=2.7 Hz, 1H), 4.82 (d, J=11.7 Hz, 1H),
4.77 (d, J=5.8 Hz, 1H), 4.74 (d, J=3.8 Hz, 1H), 4.72 (d, J=5.3 Hz,
1H), 4.68 (d, J=3.5 Hz, 1H), 4.63 (d, J=4.1 Hz, 1H), 4.51 (bs, 1H),
4.49 (t, J=3.8 Hz, 2H), 4.46 (d, J=3.5 Hz, 1H), 4.39 (d, J=10.6 Hz,
1H), 4.33-4.30 (m, 5H), 4.28-4.25 (m, 3H), 4.24-4.21 (m, 4H),
4.21-4.16 (m, 6H), 4.16-4.11 (m, 5H), 4.05 (q, J=3.7 Hz, 2H),
4.01-4.0 (m, 3H), 3.98-3.93 (m, 3H), 3.87-3.82 (m, 2H), 3.81-3.76
(m, 3H), 3.75 (s, 3H), 3.74 (d, J=6.1 Hz, 1H), 3.68 (d, J=9.1 Hz,
1H), 3.66 (d, J=9.0 Hz, 1H), 3.64 (bs, 1H), 3.63-3.57 (m, 1H),
3.57-3.54 (m, 1H), 3.53-3.45 (m, 2H), 3.42 (t, J=9.8 Hz, 2H), 3.35
(d, J=3.8 Hz, 1H), 3.34 (q, J=2.2 Hz, 2H), 3.32 (d, J=3.7 Hz, 1H),
3.30 (d, J=4.0 Hz, 1H), 3.28 (d, J=3.6 Hz, 1H), 3.26 (d, J=3.7 Hz,
1H), 3.25-3.18 (m, 1H), 2.01 (s, 3H), 2.01 (s, 6H), 2.01 (s, 3H),
1.98 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6,
170.5, 165.8, 165.5, 155.2, 150.4, 137.7, 137.4, 137.3, 133.3,
130.8, 129.8, 129.7, 129.6, 128.6, 128.5, 128.4, 128.2, 128.0,
127.9, 127.8, 118.4, 114.7, 98.2, 97.9, 97.4, 97.3, 80.6, 79.3,
79.2, 77.9, 77.4, 76.8, 75.2, 75.1, 75.0, 74.6, 74.2, 74.0, 73.9,
73.8, 73.3, 73.2, 73.15, 73.1, 72.9, 72.3, 72.1, 70.5, 70.1, 70.0,
69.5, 69.2, 69.1, 68.9, 68.0, 67.9, 67.8, 67.7, 64.1, 64.0, 63.8,
62.8, 62.7, 62.4, 61.6, 61.2, 61.1, 55.6, 20.8.
[0414] Synthesis of 49
[0415] Compound 49 is prepared from compound 41 following general
procedure F: 820 mg, 95%, R.sub.f=0.1 (Toluene/EtOAc 7:3).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.4, 165.0,
155.6, 151.1, 138.24, 138.2, 138.17, 137.9, 137.6, 137.44, 137.4,
133.7, 133.6, 133.4, 129.8, 129.6, 129.1, 129.0, 128.7, 128.6,
128.5, 128.3, 128.2, 128.1, 128.0, 128.0, 127.9, 127.7, 127.6,
127.2, 127.1, 118.4, 114.6, 101.1, 101.0, 100.9, 100.4, 97.6, 97.3,
97.2, 83.5, 83.4, 83.35, 80.3, 78.0, 77.9, 77.8, 77.7, 77.6, 75.6,
75.2, 75.1, 75.0, 74.9, 74.8, 74.5, 74.4, 73.9, 73.5, 73.4, 72.3,
70.0, 69.6, 69.5, 63.2, 62.9, 62.8, 62.7, 62.6, 62.2, 62.0, 61.8,
61.5, 61.4, 61.3, 55.6, 20.8
[0416] Synthesis of 50
[0417] Compound 50 is prepared from compound 44 following general
procedure F: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 3:2) to furnish the disaccharide as a foam,
115 mg, 83% yield, Rf=0.25 (EtOAc:petroleum ether, 3:2). HRMS (ESI)
calcd for C.sub.189H.sub.199N.sub.15O.sub.57Na(M+Na).sup.+ m/z
3613.3085, found 3613.3083. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (dd, J=6.4 Hz, J=9.1 Hz, 2 H), 8.10-8.07 (m, 6 H),
7.47-7.44 (m, 4 H), 7.43-7.40 (m, 6 H), 7.40-7.36 (m, 10H),
7.37-7.32 (m, 8 H), 7.32-7.30 (m, 12H), 7.30-7.25 (m, 12H),
7.24-7.21 (m, 8H), 7.20-7.16 (m, 6H), 7.16-7.14 (m, 6H), 7.07 (d,
J=9.1 Hz, 2H), 6.84 (d, J=9.1 Hz, 2H), 5.65 (d, J=4.0 Hz, 1H), 5.37
(t, J=3.1 Hz, 2H), 5.15-5.10 (m, 6H), 5.06-5.02 (m, 4H), 5.02 (d,
J=11.1 Hz, 1H), 4.88-4.83 (m, 3H), 4.82 (d, J=12.3 Hz, 1H), 4.77
(d, J=4.6 Hz, 1H), 4.74-4.70 (m, 10H), 4.68-4.65 (m, 12H), 4.63 (d,
J=3.8 Hz, 1H), 4.51 (bs, 1H), 4.49-4.43 (m, 6H), 4.39 (d, J=10.3
Hz, 1H), 4.33-4.30 (m, 6H), 4.26-4.21 (m, 3H), 4.21-4.17 (m, 6H),
4.16-4.10 (m, 3H), 4.05-4.0 (m, 3H), 3.99-3.94 (m, 3H), 3.87-3.82
(m, 2H), 3.81-3.77 (m, 3H), 3.76 (s, 3H), 3.75-3.72 (m, 1H), 3.68
(d, J=9.0 Hz, 1H), 3.66 (d, J=9.0 Hz, 1H), 3.62-3.57 (m, 1H),
3.57-3.53 (m, 1H), 3.52-3.45 (m, 2H), 3.41 (t, J=8.1 Hz, 2H), 3.35
(d, J=3.8 Hz, 1H), 3.34 (d, J=3.6 Hz, 1H), 3.32 (d, J=3.7 Hz, 1H),
3.30 (dd, J=4.0 Hz, J=9.0 Hz, 2H), 3.29 (t, J=5.6 Hz, 2H), 3.27 (d,
J=3.8 Hz, 1H), 3.24-3.18 (m, 1H), 2.009 (s, 6H), 2.004 (s, 6H),
1.98 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6,
170.5, 165.9, 165.8, 165.7, 155.3, 150.4, 137.7, 137.6, 137.5,
137.4, 133.4, 133.3, 130.9, 129.9, 129.84, 129.8, 129.7, 129.66,
129.6, 128.6, 128.58, 128.57, 128.5, 128.47, 128.4, 128.23, 128.22,
128.18, 128.14, 128.1, 128.0, 127.9, 127.93, 127.9, 127.8, 118.4,
114.7, 98.2, 98.1 (3C), 97.9 (3C), 97.8, 97.7, 97.3, 80.6, 79.3,
79.2, 77.8, 77.4, 77.1, 76.9, 75.2, 75.1, 75.0, 74.6, 74.2, 74.0,
73.9, 73.8, 73.5, 73.4, 73.3, 73.2, 73.1, 73.0, 72.9, 72.3, 72.1,
70.5, 70.1, 70.05, 70.0, 69.5, 69.2, 69.1, 68.9, 68.0, 67.9, 67.8,
67.7, 64.1, 64.0, 63.8, 62.8, 62.4, 61.6, 61.2, 61.1, 61.0, 55.7,
20.8.
[0418] Synthesis of 139
[0419] Compound 139 is prepared from compound 138 following general
procedure F: The residue is purified by silica gel chromatography
(EtOAc:toluene, 3:2) to furnish the disaccharide as a foam (518 mg,
121 .mu.mol, 80% yield); TLC (EtOAc:petroleum ether, 3:2, v/v)
R.sub.f=0.22; HRMS (ESI) calcd for
C.sub.224H.sub.236N.sub.18O.sub.68Na.sub.2(M+2Na).sup.2+ m/z
2156.7711, found 2156.7732. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.16 (dd, J=6.0 Hz, J=9.7 Hz, 2 H), 8.12-8.08 (m, 6 H),
7.47 (d, J=7.2 Hz, 2 H), 7.43-7.38 (m, 12 H), 7.37-7.31 (m, 14H),
7.31-7.24 (m, 18 H), 7.24-7.21 (m, 22H), 7.21-7.16 (m, 14H),
7.16-7.10 (m, 5H), 7.07 (d, J=9.0 Hz, 2H), 6.84 (d, J=9.0 Hz, 2H),
5.64 (d, J=4.0 Hz, 1H), 5.37 (t, J=3.3 Hz, 2H), 5.17-5.11 (m, 6H),
5.08-5.05 (m, 4H), 5.04-5.02 (m, 1H), 5.01 (d, J=11.0 Hz, 1H),
4.89-4.83 (m, 5H), 4.82 (d, J=11.7 Hz, 1H), 4.77-4.76 (m, 1H),
4.75-4.73 (m, 8H), 4.73-4.71 (m, 6H), 4.71-4.69 (m, 4H), 4.69-4.65
(m, 12H), 4.62 (d, J=3.9 Hz, 1H), 4.52-4.43 (m, 8H), 4.40 (d,
J=11.0 Hz, 1H), 4.35-4.23 (m, 7H), 4.23-4.12 (m, 8H), 4.07-4.01 (m,
4H), 3.98 (d, J=10.3 Hz, 1H), 3.88-3.79 (m, 6H), 3.79-3.73 (m, 3H),
3.72 (s, 3H), 3.69-3.66 (m, 2H), 3.65-3.58 (m, 2H), 3.58-3.55 (m,
2H), 3.54-3.46 (m, 4H), 3.41 (t, J=9.1 Hz, 2H), 3.35-3.29 (m, 4H),
3.29-3.27 (m, 2H), 3.27-3.20 (m, 5H), 2.0 (s, 12H), 1.99 (s, 3H),
1.97 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6,
170.5, 165.9, 165.8, 165.7, 155.3, 150.5, 137.9, 137.8, 137.6,
137.5, 137.4, 133.4, 130.0, 129.9, 129.7, 129.1, 128.6, 128.5,
128.4, 128.3, 128.2, 128.1, 127.9, 127.8, 125.4, 118.4, 114.7,
98.2, 97.9, 97.8, 97.4, 80.7, 79.4, 79.3, 77.0, 75.2, 75.1, 74.7,
74.3, 74.1, 73.9, 73.8, 73.4, 73.2, 73.0, 72.9, 72.3, 72.1, 70.5,
70.2, 69.6, 69.3, 69.2, 68.9, 68.1, 67.9, 67.8, 64.2, 64.1, 63.8,
62.8, 62.5, 61.6, 61.2, 55.7, 20.8, 20.7.
[0420] Synthesis of 51
[0421] Compound 51 is prepared from compound 45 following general
procedure F: 760 mg, 92%, R.sub.f=0.07 (Toluene/EtOAc 7:3).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.5, 170.45,
170.4, 165.0, 155.6, 151.1, 138.3, 138.2, 138.1, 137.9, 137.6,
137.4, 137.3, 133.7, 133.6, 133.4, 129.8, 129.6, 129.1, 129.0,
128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 128.0, 128.0,
127.9, 127.8, 127.7, 127.6, 127.2, 127.1, 118.4, 114.6, 101.1,
101.0, 100.9 (2C), 100.4, 97.6, 97.3 (3C), 97.2, 83.5, 83.4, 83.35,
80.3, 78.0, 77.9, 77.8, 77.7, 77.6, 77.5, 75.6, 75.3, 75.2, 75.1,
75.0, 74.9, 74.8, 74.6, 74.5, 74.4, 73.9, 73.5, 73.45, 73.4, 72.3,
70.0, 69.6, 69.5, 63.3, 62.9, 62.8, 62.7, 62.6, 62.2, 62.0, 61.8,
61.5, 61.4, 61.3, 55.6, 20.8
[0422] General Procedure G1 (GPG1): TEMPO/BAIB Oxidation and
Esterification by Diazomethane.
[0423] A solution of starting material in acetonitrile (5 mL for 32
.mu.mol) and water (0.9 mL) is treated with TEMPO (0.2 equiv.) and
BAIB (2.5 equiv. per hydroxyl group) at room temperature for 4-24
hours. After TLC (EtOAc:petroleum ether, 3:2) indicates the
completion of the reaction chloroform and water are added. The
solution is acidified with diluted HCl, back-extracted with
chloroform, dried and concentrated. The residue is dissolved in dry
ether and treated with an excess of freshly prepared diazomethane
solution in ether until TLC (EtOAc:petroleum ether, 2:3) indicates
the formation of methyl esters. The residues are purified by silica
gel chromatography (EtOAc:petroleum ether, 2:3) to furnish the
esters.
[0424] General Procedure G2 (GPG2): TEMPO/BAIB Oxidation and
Esterification with TMS-Diazomethane.
[0425] A solution of starting material in acetonitrile (5 mL for 32
.mu.mol) and water (0.9 mL) is treated with TEMPO (0.2 equiv.) and
BAIB (2.5 equiv. per hydroxyl group) at room temperature for 4-24
hours. After TLC (EtOAc:petroleum ether, 3:2) indicated the
completion of the reaction chloroform and water are added. The
solution is acidified with diluted HCl, back-extracted with
chloroform, dried and concentrated. The residue is dissolved in
diethyl ether/methanol (3:2) and and a 2M solution of
TMS-diazomethane in hexane (1.5 eq per carboxylate) is added
dropwise at 0.degree. C. After completion (TLC:Tol/EtOAc 3:2), 0.5
mL acetic acid are added to quench the reaction. Solvents are
evaporated in vacuo and the residues are purified by silica gel
chromatography to furnish the esters.
[0426] General Procedure G3 (GPG3): TEMPO/BAIB Oxidation and
Esterification with Iodomethane
[0427] A solution of starting material in acetonitrile (5 mL for 32
.mu.mol) and water (0.9 mL) is treated with TEMPO (0.2 equiv.) and
BAIB (2.5 equiv. per hydroxyl group) at room temperature for 4-24
hours. After TLC (EtOAc:petroleum ether, 3:2) indicates the
completion of the reaction chloroform and water are added. The
solution is acidified with diluted HCl, back-extracted with
chloroform, dried and concentrated. The residue is dissolved in
anhydrous DMF (100 mL per mmol). Potassium bicarbonate (20 eq) and
iodomethane (15 eq) are added and the mixture is stirred at ambient
temperature for 12-24 h. The reaction mixture is concentrated and
partitioned between ethyl acetate and water, washed with brine,
dried over magnesium sulfate and concentrated. Purification on
silica gel furnishes the esters.
[0428] Synthesis of 52
[0429] Compound 52 is prepared from compound 46 following general
procedure G1: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:2) to furnish the disaccharide as a foam,
280 mg, 82% yield, R.sub.f=0.75 (EtOAc:petroleum ether, 1:1). HRMS
(ESI) calcd for C.sub.122H.sub.125N.sub.9O.sub.38Na(M+Na).sup.+ m/z
2346.8023, found 2346.8005. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.13-8.10 (m, 4 H), 8.09 (dd, J=7.1 Hz, J=9.3 Hz, 2H),
7.55-7.47 (m, 4 H), 7.47-7.40 (m, 6 H), 7.37-7.32 (m, 4H),
7.31-7.28 (m, 4 H), 7.28-7.26 (m, 6H), 7.25-7.22 (m, 8H), 7.18-7.15
(m, 6H), 7.15-7.12 (m, 6H), 7.03 (d, J=8.7 Hz, 2H), 6.81 (d, J=9.4
Hz, 2H), 5.72 (d, J=3.9 Hz, 1H), 5.49 (d, J=4.9 Hz, 1H), 5.45 (d,
J=5.4 Hz, 1H), 5.32 (t, J=2.3 Hz, 2H), 5.19-5.14 (m, 1H), 4.99 (d,
J=2.4 Hz, 1H), 4.97 (t, J=4.0 Hz, 2H), 4.91 (d, J=2.6 Hz, 1H), 4.89
(d, J=4.0 Hz, 1H), 4.81-4.78 (m, 1H), 4.78-4.75 (m, 1H), 4.74 (d,
J=3.6 Hz, 1H), 4.72 (d, J=3.9 Hz, 1H), 4.66 (d, J=3.9 Hz, 1H), 4.58
(d, J=10.9 Hz, 1H), 4.48 (bs, 1H), 4.47 (d, J=3.0 Hz, 1H),
4.44-4.42 (m, 1H), 4.42 (d, J=5.4 Hz, 1H), 4.32 (d, J=2.1 Hz, 1H),
4.30-4.28 (m, 1H), 4.27 (d, J=2.2 Hz, 1H), 4.25-4.23 (m, 1H), 4.23
(d, J=3.6 Hz, 2H), 4.21-4.17 (m, 2H), 4.16 (d, J=2.2 Hz, 1H), 4.14
(t, J=6.0 Hz, 2H), 4.07-4.03 (m, 1H), 3.96-3.91 (m, 1H), 3.89 (t,
J=7.8 Hz, 2H), 3.81 (d, J=3.7 Hz, 1H), 3.79-3.76 (m, 1H), 3.75 (s,
3H), 3.67 (d, J=9.6 Hz, 1H), 3.63 (s, 3H), 3.60 (d, J=8.9 Hz, 1H),
3.57 (d, J=9.1 Hz, 1H), 3.54 (s, 3H), 3.52 (d, J=9.4 Hz, 1H), 3.49
(d, J=2.8 Hz, 1H), 3.47 (d, J=2.6 Hz, 1H), 3.45 (s, 3H), 3.28 (d,
J=3.6 Hz, 1H), 3.26 (d, J=3.7 Hz, 1H), 3.25 (t, J=3.5 Hz, 2H), 3.23
(t, J=3.2 Hz, 2H), 2.077 (s, 3H), 2.073 (s, 3H), 1.97 (s, 3H);
.sup.13C-NMR (CDCl.sub.3) .delta. 170.7, 170.6, 170.5, 169.5,
169.4, 169.2, 165.6, 165.2, 155.3, 150.5, 137.9, 137.8, 137.7,
137.6, 137.55, 137.5, 137.4, 137.3, 133.6, 133.5, 129.96, 129.9,
129.5, 129.2, 129.0, 128.8, 128.7, 128.5, 128.4, 128.2, 128.1,
128.0, 127.9, 127.8, 127.5, 125.3, 117.9, 114.7, 99.1, 98.9, 98.6,
98.5, 98.4, 98.2, 98.1, 79.9, 78.4, 78.3, 77.5, 77.3, 77.0, 75.9,
75.5, 75.4, 75.0, 74.8, 74.7, 74.4, 74.3, 74.1, 72.9, 72.5, 71.5,
69.8, 69.7, 68.1, 63.5, 63.4, 63.2, 62.3, 61.7, 55.7, 52.1, 52.0,
51.7, 21.4, 20.8.
[0430] Synthesis of 53
[0431] Compound 53 is prepared from compound 47 following general
procedure G3: 179 mg, 75%, R.sub.f=0.55 (Toluene/EtOAc 4:1).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.5, 168.1,
167.7, 167.6, 165.0, 164.7, 155.7, 151.0, 138.2, 138.1, 137.9,
137.6, 137.5, 137.3, 137.2, 137.1, 133.7, 133.5, 129.95, 129.9,
129.8, 129.4, 129.1, 129.0, 128.9, 128.8, 128.6, 128.4, 128.3,
128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 127.5, 118.7, 114.5,
101.11, 101.1, 100.7, 97.7, 97.3, 97.2, 82.9, 82.6, 82.2, 80.2,
78.0, 77.8, 77.6, 77.5, 75.6, 75.3, 75.0, 74.95, 74.9, 74.8, 74.7,
74.5, 74.4, 74.3, 74.1, 73.8, 73.6, 73.5, 69.9, 69.15, 69.1, 63.4,
62.8, 62.2, 61.55, 61.5, 55.6, 52.7, 52.1, 52.0, 20.9, 20.8
[0432] Synthesis of 54
[0433] Compound 54 is prepared from compound 48 following general
procedure G1: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 2:3) to furnish the disaccharide as a foam,
72 mg, 80% yield, R.sub.f=0.5 (EtOAc:petroleum ether, 2:3). HRMS
(ESI) calcd for C.sub.158H.sub.162N.sub.12O.sub.50Na(M+Na).sup.+
m/z 3050.04, found 3050.0403. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.14-8.06 (m, 8 H), 7.56-7.48 (m, 4 H), 7.48-7.43 (m, 6 H),
7.43-7.39 (m, 10H), 7.38-7.33 (m, 12H), 7.33-7.28 (m, 5 H),
7.28-7.22 (m, 6H), 7.22-7.17 (m, 8H), 7.16-7.10 (m, 6H), 7.03 (d,
J=8.7 Hz, 2H), 6.81 (d, J=9.3 Hz, 2H), 5.71 (d, J=3.9 Hz, 1H), 5.50
(d, J=5.1 Hz, 1H), 5.47 (d, J=6.0 Hz, 1H), 5.45 (d, J=6.0 Hz, 1H),
5.31 (t, J=2.4 Hz, 2H), 5.19-5.14 (m, 2H), 4.98 (bs, 1H), 4.97-4.94
(m, 3H), 4.92 (d, J=3.9 Hz, 1H), 4.90-4.88 (m, 3H), 4.82-4.74 (m,
4H), 4.72-4.67 (m, 2H), 4.66 (d, J=5.2 Hz, 1H), 4.58 (d, J=11.1 Hz,
1H), 4.49-4.46 (m, 2H), 4.45-4.40 (m, 2H), 4.36 (d, J=5.9 Hz, 1H),
4.32-4.30 (m, 5H), 4.29-4.21 (m, 3H), 4.21-4.15 (m, 4H), 4.15-4.10
(m, 3H), 4.09-4.02 (m, 2H), 3.96-3.92 (m, 2H), 3.91 (d, J=6.3 Hz,
1H), 3.88-3.83 (m, 2H), 3.80-3.72 (m, 2H), 3.75 (s, 3H), 3.68 (d,
J=10.5 Hz, 1H), 3.63 (s, 3H), 3.61 (d, J=9.8 Hz, 1H), 3.57 (d,
J=5.6 Hz, 1H), 3.53 (s, 3H), 3.52 (d, J=7.0 Hz, 1H), 3.48 (s, 3H),
3.47-3.45 (m, 2H), 3.44 (s, 3H), 3.30-3.27 (m, 1H), 3.27 (d, J=2.3
Hz, 1H), 3.26-3.24 (m, 2H), 3.24-3.22 (m, 2H), 3.21 (d, J=3.4 Hz,
1H), 2.07 (s, 3H), 2.06 (s, 6H), 1.96 (s, 3H); .sup.13C-NMR (125
MHz, CDCl.sub.3) .delta. 170.67, 170.6, 170.5, 169.5, 169.4, 169.2,
165.6, 165.2, 155.3, 150.5, 137.8, 137.7, 137.6, 137.3, 137.2,
133.7, 133.6, 129.96, 129.9, 129.86, 129.5, 129.2, 129.1, 128.8,
128.7, 128.5, 128.47, 128.4, 128.37, 128.3, 128.2, 128.1, 127.9,
127.8, 127.9, 127.8, 127.7, 127.6, 127.5, 117.9, 114.7, 99.1, 98.9,
98.7, 98.6, 98.3, 98.2, 98.1, 98.0, 79.9, 78.4, 77.5, 77.3, 77.1,
76.8, 76.2, 75.9, 75.8, 75.6, 75.1, 75.0, 74.9, 74.7, 74.6, 74.3,
74.1, 72.8, 72.5, 71.7. 71.1, 70.9, 70.5, 70.1, 69.8, 69.7, 68.1,
63.5, 63.4, 63.2, 63.1, 63.0, 62.3, 61.8, 61.6, 55.7, 52.1, 52.0,
20.7.
[0434] Synthesis of 55
[0435] Compound 55 is prepared from compound 49 following general
procedure G2: 426 mg, 59%, R.sub.f=0.5 (Toluene/EtOAc 4:1).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.65, 170.6, 168.1,
167.8, 167.6, 165.1, 164.75, 164.7, 155.8, 151.0, 138.2, 138.1,
137.9, 137.7, 137.6, 137.3, 137.2, 137.1, 133.8, 133.5, 129.9,
129.8, 129.5, 129.1, 129.0, 128.9, 128.8, 128.6, 128.4, 128.3,
128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 118.8, 114.6, 101.2,
100.8, 97.7, 97.4, 97.3, 82.9, 82.6, 82.2, 80.3, 77.9, 75.6, 75.3,
75.1, 75.0, 74.7, 74.6, 74.5, 74.3, 74.1, 73.8, 73.6, 70.0, 69.2,
63.5, 62.8, 62.3, 61.5, 55.6, 52.8, 52.1, 52.0, 20.9
[0436] Synthesis of 56
[0437] Compound 56 is prepared from compound 50 following general
procedure G1: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 2:3) to furnish the disaccharide as a foam,
92 mg, 77% yield, R.sub.f=0.45 (EtOAc:petroleum ether, 2:3). HRMS
(ESI) calcd for C.sub.194H.sub.199N.sub.15O.sub.62Na(M+Na).sup.+
m/z 3753.284, found 3753.281. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.13-8.07 (m, 10 H), 7.57-7.52 (m, 4 H), 7.52 (d, J=8.3 Hz,
1H), 7.49-7.43 (m, 10 H), 7.43-7.39 (m, 8H), 7.38-7.33 (m, 14H),
7.33-7.28 (m, 10 H), 7.28-7.23 (m, 12H), 7.22-7.16 (m, 6H),
7.15-7.10 (m, 4H), 7.03 (d, J=8.9 Hz, 2H), 6.81 (d, J=9.4 Hz, 2H),
5.71 (d, J=3.9 Hz, 1H), 5.50 (t, J=4.4 Hz, 2H), 5.47 (d, J=2.9 Hz,
1H), 5.45 (t, J=5.6 Hz, 2H), 5.32 (t, J=3.2 Hz, 2H), 5.20-5.13 (m,
2H), 4.99-4.94 (m, 2H), 4.93-4.88 (m, 2H), 4.82-4.79 (m, 4H),
4.79-4.75 (m, 2H), 4.75 (d, J=3.9 Hz, 1H), 4.72 (d, J=4.1 Hz, 1H),
4.69-4.67 (m, 4H), 4.67-4.64 (m, 4H), 4.58 (d, J=11.1 Hz, 1H),
4.50-4.46 (m, 3 H), 4.45-4.41 (m, 2H), 4.40 (d, J=5.9 Hz, 1H), 4.36
(d, J=5.3 Hz, 1H), 4.32-4.30 (m, 3H), 4.30-4.27 (m, 5H), 4.27-4.21
(m, 4H), 4.21-4.15 (m, 2H), 4.15-4.11 (m, 3H), 4.11-4.08 (m, 3H),
4.08-4.03 (m, 2H), 3.97-3.90 (m, 2H), 3.89-3.82 (m, 2H), 3.80-3.77
(m, 2H), 3.75 (s, 3H), 3.74-3.70 (m, 2H), 3.68-3.65 (m, 2H), 3.63
(s, 3H), 3.61 (d, J=9.9 Hz, 1H), 3.57 (d, J=2.0 Hz, 1H), 3.53 (s,
3H), 3.51-3.50 (m, 1H), 3.48 (s, 3H), 3.46 (s, 3H), 3.43 (s, 3H),
3.30 (t, J=3.5 Hz, 2H), 3.28-3.26 (m, 1H), 3.26 (t, J=3.3 Hz, 2H),
3.24 (d, J=3.6 Hz, 1H), 3.22 (d, J=3.8 Hz, 1H), 2.07 (s, 6H), 2.06
(s, 6H), 1.96 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.7, 170.64, 170.6, 170.5, 169.6, 169.55, 169.5, 169.2, 165.6,
165.22, 165.2, 165.19, 165.17, 155.3, 150.5, 137.8, 137.7, 137.66,
137.6, 137.5, 137.4, 137.3, 137.1, 133.8, 133.6, 133.5, 129.9,
129.8, 129.5, 129.2, 129.1, 128.8, 128.7, 128.5, 128.49, 128.4,
128.39, 128.37, 128.3, 128.2, 128.15, 128.1, 128.0, 127.95, 127.9,
127.8, 127.7, 127.6, 127.5, 117.97, 114.7, 99.1, 98.9, 98.8, 98.7,
98.6, 98.4, 98.2, 98.1, 98.03, 98.01, 80.0, 78.4, 78.1, 77.7, 76.2,
75.9, 75.8, 75.6, 75.5, 75.1, 75.0, 74.9, 74.7, 74.6, 74.4, 74.1,
72.9, 71.8, 71.7, 71.6, 71.3, 71.2, 70.9, 70.6, 70.1, 69.8, 69.7,
68.1, 63.2, 61.8, 61.7, 55.7, 52.1, 52.0, 51.8, 20.8.
[0438] Synthesis of 140
[0439] Compound 140 is prepared from compound 139 following general
procedure G1: The residue is purified by silica gel chromatography
(EtOAc:petroleum ether, 1:1) to furnish the disaccharide as a foam
(325 mg, 73.3 .mu.mol, 76% yield); TLC (EtOAc:petroleum ether, 2:3,
v/v): R.sub.f=0.41; HRMS (ESI) calcd for
C.sub.230H.sub.236N.sub.18O.sub.74Na.sub.2 (M+2Na).sup.2+ m/z
2241.2573, found 2241.2549. .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. 8.14-8.06 (m, 10 H), 7.57-7.52 (m, 4 H), 7.52 (d, J=7.3 Hz,
2H), 7.49-7.43 (m, 10 H), 7.43-7.40 (m, 8H), 7.37-7.33 (m, 14H),
7.32-7.27 (m, 18 H), 7.26-7.21 (m, 14H), 7.21-7.15 (m, 10H),
7.15-7.11 (m, 5H), 7.03 (d, J=9.1 Hz, 2H), 6.81 (d, J=9.1 Hz, 2H),
5.72 (d, J=3.8 Hz, 1H), 5.51-5.41 (m, 4H), 5.32 (t, J=3.2 Hz, 2H),
5.20-5.14 (m, 3H), 4.99-4.95 (m, 3H), 4.94-4.87 (m, 5H), 4.84-4.73
(m, 6H), 4.72-4.68 (m, 8H), 4.67-4.64 (m, 4H), 4.58 (d, J=10.8 Hz,
1H), 4.50-4.41 (m, 5H), 4.40-4.37 (m, 6H), 4.36 (d, J=5.0 Hz, 1H),
4.33-4.28 (m, 7H), 4.27-4.16 (m, 6H), 4.16-4.12 (m, 4H), 4.11-4.09
(m, 4H), 4.08-4.03 (m, 3H), 3.97-3.90 (m, 5H), 3.89-3.82 (m, 4H),
3.81-3.75 (m, 3H), 3.74 (s, 3H), 3.68-3.65 (m, 2H), 3.64 (s, 3H),
3.62-3.54 (m, 4H), 3.53 (s, 3H), 3.51-3.50 (m, 1H), 3.49 (s, 3H),
3.47 (s, 3H), 3.46 (s, 3H), 3.43 (s, 3H), 3.31-3.27 (m, 3H),
3.26-3.24 (m, 2H), 3.24 (d, J=3.4 Hz, 1H), 3.22 (d, J=3.6 Hz, 1H),
2.07 (s, 9H), 2.06 (s, 6H), 1.96 (s, 3H); .sup.13C-NMR (125 MHz,
CDCl.sub.3) .delta. 170.7, 170.6, 170.5, 169.6, 169.5, 169.2,
165.6, 165.2, 155.3, 150.5, 137.8, 137.7, 137.6, 137.5, 137.4,
137.3, 133.8, 133.6, 133.5, 129.9, 129.5, 129.2, 129.1, 128.8,
128.7, 128.5, 128.4, 128.3, 128.2, 128.1, 128.05, 128.01, 127.9,
127.8, 127.7, 127.6, 127.5, 117.9, 114.6, 99.1, 98.9, 98.7, 98.4,
98.2, 98.0, 80.0, 78.4, 78.1, 76.9, 76.3, 76.2, 76.1, 75.9, 75.8,
75.6, 75.5, 75.0, 74.9, 74.7, 74.4, 74.1, 72.8, 72.5, 71.8, 71.7,
71.6, 71.3, 71.2, 71.1, 70.9, 70.5, 70.1, 69.8, 69.7, 68.1, 66.6,
66.5, 63.5, 63.4, 63.2, 63.0, 62.3, 61.7, 61.6, 60.4, 55.7, 52.1,
52.0, 51.8, 20.8.
[0440] Synthesis of 57
[0441] Compound 57 is prepared from compound 51 following general
procedure G2: 426 mg, 51%, R.sub.f=0.46 (Toluene/EtOAc 4:1).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.6, 170.5, 168.0,
167.7, 167.5, 165.0, 164.65, 164.6, 155.7, 150.9, 138.1, 138.05,
138.0, 137.6, 137.5, 137.2, 137.1, 137.0, 133.7, 133.5, 129.9,
129.8, 129.4, 129.3, 129.0, 128.9, 128.8, 128.3, 128.2, 128.1,
127.9, 127.8, 127.7, 127.6, 118.7, 114.5, 101.1, 100.7, 97.7, 97.2,
97.1, 82.9, 82.6, 82.2, 80.2, 77.8, 75.6, 75.3, 75.0, 74.9, 74.8,
74.7, 74.6, 74.5, 74.4, 74.2, 74.0, 73.7, 73.5, 69.8, 69.0, 62.7,
61.5, 61.4, 61.3, 55.6, 52.7, 52.0, 51.9, 20.8
[0442] General Procedure H (GPH): Reduction of Azide Group.
[0443] Thiolacetic acid (1 mL for 20 .mu.mol) is added to the
starting material in dry pyridine (1 mL for 20 .mu.mol) at at room
temperature. The reaction mixture is stirred at room temperature
for 48-72 hours. After TLC (EtOAc:Toluene, 4:1) indicates the
completion of the reaction toluene is added and the solution is
washed with water, diluted HCl and NaHCO.sub.3 solution (sat.,
aq.), dried and concentrated. Chromatography (EtOAc:Toluene, 4:1)
affords N-acetylated products.
[0444] Synthesis of 58
[0445] Compound 58 is prepared from compound 52 following general
procedure H: The residue is purified by silica gel chromatography
(Toluene:EtOAc, 1:4) to furnish the disaccharide as a foam, 215 mg,
83% yield, Rf=0.68 (EtOAc:petroleum ether, 4:1). HRMS (ESI) calcd
for C.sub.128H.sub.137N.sub.3O.sub.41Na (M+Na).sup.+ m/z 2394.8625,
found 2394.8635. .sup.1H-NMR (500 MHz, CDCl.sub.3) 7.95-7.90 (m, 6
H), 7.49-7.42 (m, 6 H), 7.39-7.31 (m, 12H), 7.28-7.20 (m, 10 H),
7.20-7.12 (m, 10H), 7.10-7.06 (m, 6H), 6.95 (d, J=9.3 Hz, 2H), 6.73
(d, J=8.8 Hz, 2H), 5.78 (d, J=9.1 Hz, 1H), 5.70 (d, J=9.1 Hz, 1H),
5.59 (d, J=3.5 Hz, 1H), 5.28-5.25 (m, 1H), 5.24 (d, J=4.9 Hz, 1H),
5.16 (d, J=9.1 Hz, 1H), 5.12 (t, J=4.2 Hz, 2H), 5.10 (t, J=4.2 Hz,
2H), 4.89-4.85 (m, 1H), 4.84 (d, J=3.6 Hz, 1H), 4.80 (d, J=11.5 Hz,
1H), 4.74 (d, J=4.7 Hz, 1H), 4.71-4.69 (m, 1H), 4.68 (d, J=10.5 Hz,
1H), 4.63 (d, J=10.0 Hz, 1H), 4.59 (d, J=3.6 Hz, 1H), 4.55 (d,
J=11.5 Hz, 1H), 4.50 (d, J=11.0 Hz, 1H), 4.38 (t, J=11.0 Hz, 2H),
4.31-4.28 (m, 1H), 4.26-4.20 (m, 1H), 4.19 (d, J=4.3 Hz, 1H),
4.17-4.15 (m, 1H), 4.14-4.11 (m, 1H), 4.11-4.08 (m, 2H), 4.07-4.04
(m, 1H), 4.03 (m, 1H), 4.02 (t, J=3.4 Hz, 2H), 4.0 (d, J=4.6 Hz,
1H), 3.97 (d, J=4.8 Hz, 1H), 3.95 (t, J=3.5 Hz, 2H), 3.93 (t, J=4.2
Hz, 2H), 3.90-3.84 (m, 1H), 3.73 (t, J=2.3 Hz, 2H), 3.71 (t, J=3.1
Hz, 2H), 3.67 (s, 3H), 3.66-3.64 (m, 1H), 3.60-3.58 (m, 1H),
3.58-3.54 (m, 1H), 3.54-3.53 (m, 1H), 3.51 (s, 6H), 3.43-3.37 (m,
1H), 3.37-3.30 (m, 1H), 3.29 (s, 3H), 2.02 (s, 6H), 2.01 (s, 3H),
1.42 (s, 3H), 1.32 (s, 3H), 1.24 (s, 3H); .sup.13C-NMR (125 MHz,
CDCl.sub.3) .delta. 171.1, 170.9, 170.6, 170.2, 170.1, 169.9,
169.2, 169.0, 165.6, 165.4, 155.4, 150.42, 138.4, 138.3, 138.0,
137.8, 137.0, 136.8, 133.9, 133.8, 129.7, 129.0, 128.9, 128.6,
128.56, 128.5, 128.46, 128.2, 128.1, 128.0, 127.9, 127.7, 127.6,
127.4, 127.2, 118.1, 114.7, 98.4, 98.3, 98.25, 98.2, 98.1, 98.0,
97.9, 80.5, 78.2, 78.1, 77.3, 77.0, 75.2, 75.1, 74.96, 74.9, 74.7,
74.2, 73.9, 73.7, 73.6, 73.2, 72.9, 72.8, 72.7, 72.5, 71.5, 71.3,
70.5, 70.4, 70.3, 70.2, 70.1, 68.7, 62.4, 61.7, 61.6, 55.6, 52.4,
52.1, 52.0, 51.9, 51.7, 22.9, 22.7, 22.6, 21.0, 20.9, 20.7.
[0446] Synthesis of 59
[0447] Compound 59 is prepared from compound 53 following general
procedure H: 124 mg, 75%; .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.7, 170.6, 170.1, 167.6, 167.4, 155.7, 150.8, 138.7,
138.1, 137.7, 136.5, 136.4, 136.3, 133.8, 133.6, 129.8, 129.2,
129.0, 128.8, 128.6, 128.55, 128.5, 128.45, 128.3, 128.2, 128.15,
128.1, 128.0, 127.9, 127.8, 127.5, 127.4, 127.3, 118.6, 114.5,
101.2, 101.1, 100.9, 99.5, 99.0, 98.7, 81.6, 81.4, 80.8, 80.5,
78.0, 77.9, 77.8, 75.4, 75.25, 75.2, 75.15, 75.0, 74.95, 74.85,
74.8, 74.4, 74.3, 73.9, 73.7, 73.6, 70.7, 70.2, 70.1, 62.2, 61.5,
61.4, 55.6, 52.8, 52.6, 52.5, 52.1, 52.0, 22.65, 22.6, 20.8, 20.77,
20.7
[0448] Synthesis of 60
[0449] Compound 60 is prepared from compound 54 following general
procedure H: The residue is purified by silica gel chromatography
(Toluene:EtOAc, 1:4) to furnish the disaccharide as a foam, 36 mg,
70% yield, R.sub.f=0.5 (EtOAc:Toluene, 4:1). HRMS (ESI) calcd for
C.sub.166H.sub.178N.sub.4O.sub.54Na (M+Na).sup.+ m/z 3114.1203,
found 3114.1204. .sup.1H-NMR (500 MHz, CDCl.sub.3) 8.02 (d, J=10.1
Hz, 6H), 7.57-7.50 (m, 6 H), 7.47-7.39 (m, 6 H), 7.36-7.27 (m,
12H), 7.27-7.24 (m, 10 H), 7.24-7.18 (m, 10H), 7.18-7.14 (m, 6H),
7.13-7.10 (m, 6H), 7.01 (d, J=9.2 Hz, 2H), 6.80 (d, J=9.2 Hz, 2H),
5.81 (d, J=9.5 Hz, 1H), 5.78 (d, J=9.3 Hz, 1H), 5.75 (d, J=9.4 Hz,
1H), 5.66 (d, J=2.0 Hz, 1H), 5.36-5.32 (m, 1H), 5.31 (t, J=5.1 Hz,
2H), 5.21-5.15 (m, 2H), 4.95-4.92 (m, 1H), 4.92 (t, J=3.5 Hz, 2H),
4.88 (d, J=11.5 Hz, 1H), 4.81 (d, J=3.8 Hz, 1H), 4.79 (t, J=4.3 Hz,
2H), 4.75 (d, J=4.5 Hz, 1H), 4.73-4.69 (m, 3H), 4.69 (d, J=2.1 Hz,
1H), 4.66 (bs, 1H), 4.64-4.61 (m, 3H), 4.61-4.58 (m, 2H), 4.58 (d,
J=4.1 Hz, 1H), 4.55 (d, J=3.6 Hz, 1H), 4.46 (d, J=11.0 Hz, 1H),
4.41 (d, J=11.5 Hz, 1H), 4.36-4.34 (m, 4H), 4.33-4.25 (m, 4H),
4.23-4.21 (m, 5H), 4.21-4.19 (m, 2H), 4.19-4.17 (m, 2H), 4.17-4.13
(m, 2H), 4.13 (d, J=11.5 Hz, 1H), 4.09-4.05 (m, 1H), 4.05 (d, J=4.7
Hz, 1H), 4.02-4.0 (m, 1H), 4.0-3.95 (m, 1H), 3.81-3.79 (m, 1H),
3.79-3.76 (m, 1H), 3.75 (s, 3H), 3.74-3.70 (m, 1H), 3.67-3.61 (m,
1H), 3.59 (s, 3H), 3.57 (s, 3H), 3.50 (d, J=10.1 Hz, 1H), 3.47 (d,
J=3.8 Hz, 1H), 3.44 (d, J=2.4 Hz, 1H), 3.42-3.39 (m, 1H), 3.39-3.37
(m, 1H), 3.35 (s, 3H), 3.34 (s, 3H), 2.09 (s, 3H), 2.08 (s, 3H),
2.07 (s, 3H), 1.98 (s, 3H), 1.49 (s, 3H), 1.389 (s, 3H), 1.384 (s,
3H), 1.31 (s, 3H); .sup.13C-NMR (CDCl.sub.3) .delta. 170.9, 170.6,
170.1, 169.9, 169.2, 165.6, 165.4, 155.4, 150.4, 138.4, 138.0,
137.8, 137.0, 136.8, 136.4, 133.8, 129.8, 129.0, 128.9, 128.6,
128.5, 128.4, 128.3, 128.2, 128.15, 128.1, 128.0, 127.7, 127.6,
127.4, 127.2, 126.9, 118.0, 114.7, 98.4, 98.37, 98.3, 98.2, 98.15,
98.1, 97.95, 97.9, 80.5, 78.1, 77.3, 77.0, 76.8, 75.4, 75.2, 75.1,
74.96, 74.9, 74.7, 73.7, 73.2, 72.9, 72.8, 72.5, 71.6, 71.4, 70.5,
70.4, 70.2, 68.7, 68.3, 62.4, 61.5, 61.6, 55.7, 52.6, 52.4, 52.37,
52.3, 52.0, 51.7, 29.7, 29.3, 20.9, 20.7.
[0450] Synthesis of 61
[0451] Compound 61 is prepared from compound 55 following general
procedure H: 316 mg, 75%. .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.7, 170.65, 170.6, 170.2, 170.1, 167.7, 167.5, 167.4,
167.3, 165.0, 164.9, 164.8, 155,7, 150.9, 138.8, 138.7, 138.1,
137.9, 137.7, 136.5, 136.4, 136.35, 136.3, 133.9, 133.6, 129.8,
129.75, 129.2, 129.1, 128.9, 128.8, 128.65, 128.6, 128.55, 128.55,
128.5, 128. 4, 128.3, 128.25, 128.2, 128.1, 128.05, 128.0, 127.95,
127.85, 127.8, 127.55, 127.5, 127.45, 127.4, 127.35, 118.6, 114.5,
101.1, 100.9, 99.5, 99.0, 98.7, 98.6, 81.6, 81.4, 80.9, 80.5, 78.0,
77.9, 77.8, 76.7, 76.6, 75.4, 75.2, 75.1, 75.0, 74.95, 74.9, 74.8,
74.5, 74.45, 74.4, 73.8, 73.65, 73.6, 73.55, 70.7, 70.2, 70.1,
62.2, 61.5, 61.4, 55.6, 52.8, 52.7, 52.5, 52.1, 52.0, 22.7, 22.65,
22.65, 22.6, 21.5, 20.8, 20.75, 20.7
[0452] Synthesis of 62
[0453] Compound 62 is prepared from compound 56 following general
procedure H: The residue is purified by silica gel chromatography
(Toluene:EtOAc, 1:4) to furnish the disaccharide as a foam, 69 mg,
90% yield, Rf=0.5 (EtOAc:Toluene, 4:1). HRMS (ESI) calcd for
C.sub.204H.sub.219N.sub.5O.sub.67Na (M+Na).sup.+ m/z 3833.398,
found 3833.397. .sup.1H-NMR (500 MHz, CDCl.sub.3) 8.02-7.98 (m,
8H), 7.57-7.50 (m, 6 H), 7.47-7.39 (m, 8 H), 7.36-7.28 (m, 14H),
7.27-7.23 (m, 18 H), 7.23-7.14 (m, 16H), 7.13-7.09 (m, 10H), 7.01
(d, J=8.8 Hz, 2H), 6.81 (d, J=9.1 Hz, 2H), 5.84-5.79 (m, 1H), 5.79
(d, J=10.3 Hz, 1H), 5.66 (d, J=3.4 Hz, 1H), 5.40-5.35 (m, 1H),
5.35-5.32 (m, 1H), 5.30-5.26 (m, 2H), 5.24 (d, J=8.5 Hz, 1H),
5.18-5.13 (m, 2H), 4.95-4.93 (m, 1H), 4.92-4.89 (m, 2H), 4.87 (d,
J=11.4 Hz, 1H), 4.81 (d, J=4.0 Hz, 1H), 4.79-4.76 (m, 4H),
4.75-4.72 (m, 4H), 4.72-4.70 (m, 4H), 4.69-4.67 (m, 3H), 4.67-4.65
(m, 4H), 4.64-4.61 (m, 3H), 4.60-4.58 (m, 4H), 4.58-4.56 (m, 2H),
4.55 (d, J=3.6 Hz, 1H), 4.46-4.39 (m, 1H), 4.38-4.34 (m, 3H),
4.33-4.27 (m, 3H), 4.27 (d, J=4.2 Hz, 1H), 4.23-4.21 (m, 3H),
4.21-4.19 (m, 2H), 4.18-4.14 (m, 2H), 4.13-4.11 (m, 2H), 4.10-4.06
(m, 1H), 4.05 (d, J=5.1 Hz, 1H), 4.03-4.0 (m, 3H), 4.0-3.96 (m,
4H), 3.95-3.91 (m, 1H), 3.80-3.78 (m, 1H), 3.78-3.76 (m, 1H), 3.75
(s, 3H), 3.74-3.73 (m, 1H), 3.72-3.70 (m, 1H), 3.66-3.60 (m, 2H),
3.58 (s, 3H), 3.57 (s, 3H), 3.50-3.47 (m, 2H), 3.46-3.42 (m, 1H),
3.42-3.38 (m, 1H), 3.36 (s, 3H), 3.34 (s, 3H), 3.33 (s, 3H), 3.28
(d, J=6.0 Hz, 1H), 2.09 (s, 3H), 2.08 (s, 3H), 2.07 (s, 6H), 1.98
(s, 3H), 1.49 (s, 3H), 1.38 (s, 6H), 1.37 (s, 3H), 1.31 (s, 3H);
.sup.13C-NMR (CDCl.sub.3) .delta. 171.1, 170.9, 170.6, 170.2,
169.9, 169.2, 169.0, 165.6, 165.4, 165.3, 155.4, 150.4, 138.4,
138.0, 137.8, 137.1, 136.8, 136.4, 133.9, 130.9, 129.7, 129.1,
129.0, 128.9, 128.6, 128.5, 128.2, 128.1, 128.0, 127.7, 127.4,
127.2, 127.1, 127.06, 127.0, 118.1, 114.7, 98.4, 98.33, 98.3,
98.25, 98.2, 98.1, 97.96, 97.9, 97.75, 97.7, 80.5, 78.2, 78.1,
77.3, 77.1, 75.2, 75.1, 74.98, 74.9, 74.7, 74.1, 73.7, 73.6, 72.9,
72.8, 72.6, 71.7, 71.5, 71.3, 70.5, 70.4, 70.2, 68.7, 68.3, 64.7,
62.5, 61.6, 55.7, 52.4, 52.1, 51.9, 51.7, 22.9, 22.7, 20.9,
20.8.
[0454] Synthesis of 141
[0455] Compound 141 is prepared from compound 140 following general
procedure H: The residue is purified by silica gel chromatography
(Toluene:EtOAc, 1:4) to furnish the disaccharide as a clear syrup,
212 mg, 46.8 .mu.mol, 92% yield; TLC (Toluene:EtOAc, 1:4, v/v):
R.sub.f=0.45; HRMS (ESI) calcd for
C.sub.242H.sub.260N.sub.6O.sub.60Na.sub.2 (M+2Na).sup.2+ m/z
2288.8162, found 2288.8108; .sup.1H-NMR (500 MHz, CDCl.sub.3)
8.03-7.97 (m, 8H), 7.58-7.50 (m, 4 H), 7.48-7.39 (m, 5 H),
7.36-7.28 (m, 14H), 7.28-7.22 (m, 28 H), 7.22-7.14 (m, 22H),
7.14-7.1 (m, 12H), 7.02 (d, J=9.1 Hz, 2H), 6.81 (d, J=9.1 Hz, 2H),
5.88-5.79 (m, 3H), 5.67 (d, J=2.1 Hz, 1H), 5.35-5.32 (m, 2H),
5.32-5.25 (m, 3H), 5.19-5.13 (m, 3H), 4.97-4.94 (m, 1H), 4.93-4.89
(m, 2H), 4.88 (d, J=11.9 Hz, 1H), 4.82-4.76 (m, 6H), 4.75-4.70 (m,
10H), 4.70-4.68 (m, 8H), 4.68-4.62 (m, 12H), 4.61-4.58 (m, 3H),
4.58-4.54 (m, 3H), 4.47-4.43 (m, 3H), 4.43-4.35 (m, 4H), 4.34-4.25
(m, 5H), 4.25-4.23 (m, 1H), 4.22-4.19 (m, 2H), 4.19-4.15 (m, 3H),
4.15-4.12 (m, 1H), 4.11-4.06 (m, 2H), 4.06 (d, J=5.5 Hz, 1H),
4.04-4.01 (m, 2H), 4.0-3.91 (m, 5H), 3.81-3.77 (m, 2H), 3.74 (s,
3H), 3.68-3.60 (m, 4H), 3.59 (s, 3H), 3.58 (s, 3H), 3.51-3.49 (m,
1H), 3.49 (d, J=2.2 Hz, 1H), 3.47-3.45 (m, 1H), 3.45-3.43 (m, 1H),
3.43-3.38 (m, 2H), 3.37 (s, 3H), 3.35 (s, 3H), 3.34 (s, 3H), 3.33
(s, 3H), 2.1 (s, 3H), 2.09 (s, 3H), 2.08 (s, 3H), 2.07 (s, 6H),
1.98 (s, 3H), 1.49 (s, 3H), 1.39 (s, 3H), 1.38 (s, 6H), 1.37 (s,
3H), 1.31 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
171.0, 170.6, 170.2, 169.9, 169.2, 169.0, 165.6, 165.4, 155.4,
150.4, 138.4, 138.0, 137.8, 137.0, 136.8, 133.9, 130.8, 129.7,
128.9, 128.6, 128.5, 128.2, 128.0, 127.7, 127.6, 127.4, 127.2,
127.1, 127.0, 125.3, 118.1, 114.7, 98.3, 98.1, 97.9, 97.8, 80.5,
78.2, 76.9, 75.1, 74.9, 74.8, 74.6, 74.1, 73.7, 73.5, 73.2, 72.9,
72.7, 72.6, 72.5, 71.6, 71.5, 70.5, 70.4, 70.2, 68.7, 68.3, 68.2,
62.4, 61.6, 60.4, 55.6, 52.4, 52.1, 52.0, 51.8, 51.7, 22.9, 22.7,
21.5, 20.9, 20.7.
[0456] Synthesis of 63
[0457] Compound 63 is prepared from compound 57 following general
procedure H: 280 mg, 78%. .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.7, 170.6, 170.2, 167.7, 167.4, 167.35, 167.3, 165.0,
164.9, 164.8, 155.7, 150.9, 138.8, 138.1, 137.9, 137.7, 136.5,
136.4, 136.3, 133.9, 133.6, 129.8, 129.2, 129.1, 128.8, 128.7,
128.6, 128.35, 128.3, 128.2, 128.1, 128.0, 127.8, 127.5, 127.4,
118.6, 114.5, 101.2, 100.9, 99.6, 99.0, 98.75, 98.7, 81.6, 81.4,
80.8, 80.6, 78.1, 77.8, 77.6, 75.4, 75.3, 75.0, 74.9, 74.8, 74.5,
74.45, 74.4, 73.9, 73.6, 70.7, 70.2, 70.1, 62.2, 61.5, 61.4, 60.4,
55.6, 53.5, 52.9, 52.7, 52.5, 52.1, 52.0, 22.7, 21.5, 20.8
##STR00039## ##STR00040##
[0458] General Procedure I (GPI): Selective De-O-Acetylation.
[0459] Starting material is dissolved in dry dichloromethane (2 mL
for 9 .mu.mol) at 0.degree. C. and treated with a solution of cold
dry methanol (4 mL) containing 80 .mu.L of acetyl chloride.
Stirring of the reaction mixture is continued for 30 min at
0.degree. and then at 23.degree. C. for 48 h. After TLC (EtOAc)
indicates the completion of the reaction dichloromethane is added
and the solution is washed with water and NaHCO.sub.3 solution
(sat., aq.), dried and concentrated. The residues are purified by
silica gel chromatography (EtOAc:MeOH, 9:1) to give de-O-acetylated
products.
[0460] Synthesis of 64
[0461] Compound 64 is prepared from compound 58 following general
procedure I: 152 mg, 80% yield, Rf=0.45 (EtOAc). HRMS (ESI) calcd
for C.sub.122H.sub.131N.sub.3O.sub.38Na (M+Na).sup.+ m/z 2268.8308,
found 2268.8289. .sup.1H-NMR (500 MHz, CDCl.sub.3) 7.94 (d, J=7.9
Hz, 2H), 7.89 (t, J=5.4 Hz, 4H), 7.48-7.42 (m, 4 H), 7.42-7.38 (m,
6H), 7.37-7.33 (m, 6H), 7.31-7.27 (m, 6H), 7.27-7.22 (m, 8 H),
7.21-7.13 (m, 4H), 7.13-7.07 (m, 4H), 7.07-7.02 (m, 4H), 6.95 (d,
J=6.9 Hz, 2H), 6.92 (d, J=9.2 Hz, 2H), 6.71 (d, J=9.0 Hz, 2H), 5.72
(d, J=8.2 Hz, 1H), 5.61 (d, J=9.3 Hz, 1H), 5.57 (d, J=2.2 Hz, 1H),
5.35-5.32 (m, 1H), 5.30 (t, J=2.9 Hz, 2H), 5.17 (t, J=3.8 Hz, 2H),
5.12-5.07 (m, 2H), 4.91 (d, J=3.6 Hz, 1H), 4.87-4.85 (m, 2H), 4.84
(d, J=3.3 Hz, 1H), 4.78 (d, J=3.3 Hz, 1H), 4.75-4.74 (m, 3H),
4.73-4.71 (m, 4H), 4.70-4.67 (m, 2H), 4.67-4.65 (m, 2H), 4.65 (d,
J=2.9 Hz, 1H), 4.62 (d, J=11.5 Hz, 1H), 4.58 (d, J=5.0 Hz, 1H),
4.55-4.53 (m, 1H), 4.53-4.51 (m, 1H), 4.34 (q, J=11.8 Hz, 2H),
4.22-4.17 (m, 1H), 4.15-4.10 (m, 2H), 4.09 (d, J=11.8 Hz, 1H),
4.05-4.02 (m, 1H), 4.02-3.97 (m, 2H), 3.95 (t, J=4.1 Hz, 2H), 3.91
(d, J=3.7 Hz, 1H), 3.86 (t, J=3.9 Hz, 2H), 3.72-3.67 (m, 1H), 3.65
(s, 3H), 3.63-3.58 (m, 1H), 3.57-3.54 (m, 1H), 3.54-3.51 (m, 1H),
3.50 (s, 3H), 3.47 (s, 3H), 3.44-3.40 (m, 1H), 3.40-3.38 (m, 1H),
3.38 (d, J=6.5 Hz, 1H), 3.35-3.31 (m, 1H), 3.30 (s, 3H), 1.44 (s,
3H), 1.39 (s, 3H), 1.25 (s, 3H); .sup.13C-NMR (125 MHz, CDCl.sub.3)
.delta. 170.3, 169.9, 169.2, 169.1, 165.5, 165.4, 155.4, 150.5,
138.5, 138.4, 138.2, 138.1, 137.1, 136.9, 133.9, 133.8, 133.7,
129.8, 129.7, 129.6, 129.1, 129.0, 128.9, 128.6, 128.5, 128.4,
128.2, 128.1, 127.9, 127.89, 127.87, 127.82, 127.8, 127.7, 127.5,
127.2, 118.1, 114.7, 98.6, 98.2, 97.9, 97.8, 97.1, 96.9, 79.9,
77.7, 77.6, 77.4, 77.1, 74.9, 74.4, 74.2, 74.1, 73.3, 73.2, 73.0,
72.9, 72.8, 72.7, 72.6, 72.5, 72.4, 72.3, 72.2, 71.3, 70.9, 70.4,
69.8, 68.8, 61.5, 60.7, 60.5, 55.6, 52.6, 52.4, 52.2, 51.8, 29.6,
22.9, 22.7, 22.6.
[0462] Synthesis of 65
[0463] Compound 65 is prepared from compound 59 following general
procedure I: 92 mg, 90%, R.sub.f=0.35 (dichloromethane/methanol
19:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.15, 170.1,
170.0, 168.0, 167.9, 165.0, 164.95, 164.9, 155.7, 150.8, 139.0,
138.9, 138.3, 137.9, 136.5, 133.9, 133.8, 133.5, 130.9, 129.7,
129.6, 129.55, 129.3, 129.1, 128.9, 128.6, 128.5, 128.4, 128.3,
128.2, 128.15, 128.1, 128.0, 127.9, 127.8, 127.4, 127.3, 127.1,
118.6, 114.5, 100.9, 100.6, 100.5, 99.0, 98.6, 98.3, 81.4, 81.3,
81.0, 80.1, 77.8, 75.5, 75.2, 75.1, 75.0, 74.95, 74.8, 74.7, 74.2,
73.9, 73.8, 73.7, 73.3, 72.6, 72.2, 61.6, 60.1, 55.6, 52.8, 52.65,
52.6, 52.3, 52.2, 52.1, 22.7, 22.65
[0464] Synthesis of 66
[0465] Compound 66 is prepared from compound 60 following general
procedure I: 21 mg, 93% yield, R.sub.f=0.15 (EtOAc:petroleum ether,
4:1). HRMS (ESI) calcd for C.sub.158H.sub.170N.sub.4O.sub.50Na
(M+Na).sup.+ m/z 2946.0781, found 2946.0757. .sup.1H-NMR (500 MHz,
CDCl.sub.3) 8.03 (d, J=7.6 Hz, 2H), 7.98-7.93 (m, 4H), 7.57-7.49
(m, 4 H), 7.47-7.41 (m, 6H), 7.40-7.37 (m, 8H), 7.36-7.30 (m, 12H),
7.30-7.20 (m, 16H), 7.19-7.15 (m, 8H), 7.14-7.11 (m, 5H), 7.02 (d,
J=9.0 Hz, 2H), 6.81 (d, J=8.6 Hz, 2H), 5.72 (d, J=3.9 Hz, 2H), 5.70
(d, J=3.9 Hz, 2H), 5.65 (d, J=2.1 Hz, 1H), 5.62 (d, J=9.5 Hz, 1H),
5.41-5.38 (m, 1H), 5.38-5.34 (m, 1H), 5.23 (t, J=4.8 Hz, 2H),
5.20-5.16 (m, 2H), 5.16 (d, J=8.6 Hz, 2H), 4.98 (d, J=3.7 Hz, 1H),
4.95 (d, J=3.1 Hz, 1H), 4.93 (d, J=3.1 Hz, 1H), 4.86-4.83 (m, 2H),
4.81 (t, J=7.1 Hz, 2H), 4.77 (t, J=2.4 Hz, 2H), 4.74 (d, J=2.7 Hz,
1H), 4.72 (d, J=11.7 Hz, 1H), 4.68 (d, J=6.8 Hz, 1H), 4.65-4.62 (m,
6H), 4.62-4.60 (m, 6H), 4.60-4.59 (m, 4H), 4.58-4.56 (m, 4H), 4.40
(t, J=11.8 Hz, 2H), 4.29 (d, J=3.6 Hz, 1H), 4.27 (d, J=2.8 Hz, 1H),
4.25 (d, J=3.5 Hz, 1H), 4.22-4.21 (m, 1H), 4.21 (d, J=3.8 Hz, 2H),
4.18 (d, J=3.4 Hz, 1H), 4.16 (d, J=11.8 Hz, 1H), 4.09-4.03 (m, 2H),
4.01-3.95 (m, 1H), 3.93-3.89 (m, 1H), 3.75 (s, 3H), 3.70-3.65 (m,
1H), 3.65-3.61 (m, 1H), 3.59 (s, 3H), 3.57 (s, 3H), 3.56-3.53 (m,
1H), 3.52-3.50 (m, 2H), 3.49-3.45 (m, 1H), 3.44-3.42 (m, 1H),
3.44-3.38 (m, 1H), 3.37 (s, 3H), 3.36 (s, 3H), 3.35-3.32 (m, 1H),
1.46 (s, 3H), 1.44 (s, 3H), 1.33 (s, 3H), 1.26 (s, 3H);
.sup.13C-NMR (CDCl.sub.3) .delta. 170.2, 169.1, 155.4, 138.5,
138.1, 137.0, 136.8, 133.8, 129.7, 129.6, 129.0, 128.6, 128.2,
127.9, 127.5, 127.2, 125.3, 118.1, 114.7, 98.15, 98.0, 97.9, 97.8,
97.7, 97.4, 97.3, 97.2, 79.8, 77.3, 77.0, 76.8, 74.9, 74.7, 74.6,
73.2, 72.9, 72.8, 72.6, 72.5, 72.45, 72.4, 70.4, 69.9, 68.6, 68.4,
61.6, 60.6, 52.4, 52.2, 51.8, 29.7, 22.9, 22.7.
[0466] Synthesis of 67
[0467] Compound 67 is prepared from compound 61 following general
procedure I: 101 mg, 80%, R.sub.f=0.33 (dichloromethane/methanol
19:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.2, 170.15,
170.1, 167.95, 167.9, 165.1, 165.0, 164.9, 155.7, 150.8, 139.0,
138.9, 138.3, 137.9, 137.2, 136.5, 133.9, 133.8, 133.5, 129.7,
129.6, 129.5, 129.2, 129.1, 128.8, 128.6, 128.5, 128.3, 128.25,
128.2, 128.15, 128.1, 128.0, 127.9, 127.8, 127.4, 127.35, 127.3,
118.6, 114.5, 100.8, 100.6, 100.5, 100.4, 99.0, 98.6, 98.3, 98.2,
81.4, 81.3, 81.2, 81.0, 80.1, 77.8, 76.4, 76.3, 75.5, 75.15, 75.1,
75.05, 75.0, 74.95, 74.8, 74.75, 74.7, 74.2, 74.1, 73.9, 73.8,
73.3, 72.6, 72.4, 72.3, 61.6, 60.1, 60.0, 55.6, 52.8, 52.65, 52.6,
52.3, 52.15, 52.1, 22.65, 22.6
[0468] Synthesis of 68
[0469] Compound 68 is prepared from compound 62 following general
procedure I: 27 mg, 87% yield, Rf=0.41 (EtOAc). HRMS (ESI) calcd
for C.sub.194H.sub.209N.sub.5O.sub.62Na (M+Na).sup.+ m/z 3623.33,
found 3623.2315. .sup.1H-NMR (500 MHz, CDCl.sub.3) 8.03-8.0 (m,
2H), 7.97-7.92 (m, 6H), 7.58-7.52 (m, 4 H), 7.52-7.48 (m, 6H),
7.47-7.41 (m, 8H), 7.40-7.35 (m, 12H), 7.35-7.30 (m, 12H),
7.30-7.21 (m, 16H), 7.21-7.18 (m, 4H), 7.17-7.14 (m, 8H), 7.14-7.10
(m, 5H), 7.01 (d, J=9.2 Hz, 2H), 6.80 (d, J=9.2 Hz, 2H), 5.75-5.69
(m, 2H), 5.68-5.60 (m, 2H), 5.42-5.37 (m, 2H), 5.37-5.34 (m, 2H),
5.23-5.13 (m, 2H), 4.98 (d, J=3.5 Hz, 1H), 4.95 (d, J=3.0 Hz, 1H),
4.93-4.90 (m, 2H), 4.87-4.82 (m, 2H), 4.81 (d, J=11.1 Hz, 1H), 4.78
(t, J=3.9 Hz, 2H), 4.74 (d, J=2.7 Hz, 1H), 4.72-4.70 (m, 6H),
4.70-4.67 (m, 6H), 4.67 (d, J=6.8 Hz, 1H), 4.64-4.62 (m, 4H),
4.62-4.60 (m, 4H), 4.60 (d, J=4.3 Hz, 1H), 4.58 (d, J=4.3 Hz, 1H),
4.41-4.33 (m, 4H), 4.29-4.21 (m, 6H), 4.21 (d, J=4.6 Hz, 1H), 4.19
(d, J=4.6 Hz, 1H), 4.16 (d, J=11.2 Hz, 2H), 4.12-4.09 (m, 2H),
4.09-4.02 (m, 4H), 4.01-3.95 (m, 2H), 3.95-3.89 (m, 1H), 3.75 (s,
3H), 3.72-3.65 (m, 1H), 3.65-3.60 (m, 1H), 3.59 (s, 3H), 3.57 (s,
3H), 3.50-3.44 (m, 1H), 3.44-3.39 (m, 1H), 3.37 (s, 3H), 3.35 (s,
3H), 3.34 (s, 3H), 1.46 (s, 3H), 1.45 (s, 3H), 1.43 (s, 3H), 1.33
(s, 3H), 1.25 (s, 3H); .sup.13C-NMR (CDCl.sub.3) .delta. 170.3,
169.9, 169.2, 169.1, 165.5, 165.4, 155.4, 138.5, 138.2, 138.1,
137.1, 136.8, 133.9, 129.8, 129.6, 129.0, 128.6, 128.5, 128.2,
127.9, 127.5, 127.4, 127.2, 118.1, 114.7, 98.6, 98.5, 98.3, 98.2,
98.1, 97.8, 97.7, 97.1, 97.0, 79.9, 77.6, 77.3, 77.0, 76.8, 74.9,
74.8, 74.6, 73.4, 72.9, 72.6, 72.5, 72.4, 72.3, 70.9, 70.2, 68.7,
68.4, 60.6, 55.7, 52.6, 52.1, 51.8, 29.7, 22.9, 22.7.
[0470] Synthesis of 142
[0471] Compound 142 is prepared from compound 141 following general
procedure I: 142 mg, 33.2 .mu.mol, 100% yield, TLC (EtOAc):
R.sub.f=0.38; HRMS (ESI) calcd for
C.sub.230H.sub.248N.sub.6O.sub.74Na.sub.2 (M+2Na).sup.2+ m/z
2162.7844, found 2162.7808. .sup.1H-NMR (500 MHz, CDCl.sub.3)
8.02-7.99 (m, 2H), 7.97-7.91 (m, 8H), 7.58-7.50 (m, 6 H), 7.47-7.40
(m, 12H), 7.39-7.31 (m, 18H), 7.30-7.17 (m, 28H), 7.17-7.09 (m,
16H), 7.03-7.0 (m, 5H), 7.0 (d, J=9.0 Hz, 2H), 6.79 (d, J=9.0 Hz,
2H), 5.96-5.81 (m, 2H), 5.66 (d, J=2.3 Hz, 1H), 5.44-5.40 (m, 2H),
5.40-5.35 (m, 2H), 5.29 (d, J=8.8 Hz, 1H), 5.24-5.20 (m, 1H),
5.20-5.13 (m, 2H), 5.0-4.89 (m, 3H), 4.88 (d, J=3.7 Hz, 1H), 4.85
(d, J=11.2 Hz, 1H), 4.81-4.75 (m, 10H), 4.75-4.72 (m, 8H),
4.72-4.66 (m, 10H), 4.66-4.64 (m, 6H), 4.64-4.56 (m, 10H),
4.42-4.34 (m, 8H), 4.32-4.23 (m, 6H), 4.23-4.14 (m, 5H), 4.11-4.05
(m, 6H), 4.03-3.97 (m, 4H), 3.96-3.90 (m, 8H), 3.74 (s, 3H),
3.70-3.62 (m, 4H), 3.59 (s, 3H), 3.54 (s, 3H), 3.50-3.44 (m, 2H),
3.44-3.38 (m, 2H), 3.36 (s, 3H), 3.35 (s, 6H), 3.33 (s, 3H), 1.45
(s, 3H), 1.43 (s, 6H), 1.42 (s, 3H), 1.32 (s, 6H); .sup.13C-NMR
(125 MHz, CDCl.sub.3) .delta. 170.5, 170.4, 170.0, 169.3, 169.1,
169.0, 165.5, 165.4, 155.4, 150.4, 138.5, 138.2, 138.1, 137.4,
137.1, 136.9, 136.8, 133.9, 129.7, 129.6, 129.1, 129.0, 128.6,
128.4, 128.2, 128.1, 127.9, 127.8, 127.7, 127.5, 127.4, 127.2,
118.1, 114.7, 98.5, 98.2, 98.1, 97.7, 96.9, 96.8, 96.7, 80.0, 77.2,
77.0, 76.9, 76.7, 76.4, 76.2, 75.1, 74.9, 74.8, 74.7, 74.5, 74.4,
74.1, 73.9, 73.8, 73.3, 73.2, 73.0, 72.8, 72.7, 72.6, 72.5, 72.4,
72.1, 72.0, 71.2, 71.0, 70.8, 70.2, 69.9, 69.7, 68.8, 68.6, 68.3,
67.4, 67.1, 66.7, 66.5, 66.4, 66.3, 66.2, 61.4, 60.5, 55.6, 52.6,
52.4, 52.2, 51.8, 22.9, 22.7, 22.6, 20.9.
[0472] Synthesis of 69
[0473] Compound 69 is prepared from compound 63 following general
procedure I: 120 mg, 91%, R.sub.f=0.3 (dichloromethane/methanol
19:1). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 170.2, 170.1,
170.0, 167.9, 165.1, 165.0, 164.9, 155.7, 150.9, 139.0, 138.3,
137.9, 137.2, 136.5, 133.9, 133.8, 133.6, 129.7, 129.6, 129.3,
129.1, 128.9, 128.6, 128.5, 128.3, 128.25, 128.2, 128.1, 128.0,
127.9, 127.8, 127.4, 118.6, 114.5, 100.9, 100.6, 100.5, 100.4,
99.0, 98.6, 98.3, 98.2, 81.4, 81.3, 81.2, 81.1, 80.1, 77.8, 76.7,
76.4, 76.3, 75.5, 75.2, 75.1, 75.0, 74.8, 74.7, 74.4, 74.2, 74.1,
74.0, 73.8, 73.3, 72.6, 72.4, 72.35, 61.6, 60.1, 60.0, 55.6, 52.8,
52.65, 52.6, 52.3, 52.1, 22.7, 22.65
[0474] General Procedure J (GPJ): O-Sulfation.
[0475] Sulfur trioxide trimethylamine complex (5 equiv per hydroxyl
group) is added to the starting materials in dry DMF (3 mL for 50
mg). The mixture is heated at 50-60.degree. C. under argon for
48-72 h. MeOH (1 mL) is added and the mixture stirred for 15 min
and concentrated in vacuo. Chromatography
(dichloromethane:methanol:aq. ammonia, 7:2:0.5) affords O-sulfated
products.
[0476] General Procedure K (GPK): Saponification.
[0477] Starting material is dissolved in methanol and water (4/1,
v/v, 1.25 mL for 20 mg) containing 2M solution of sodium hydroxide
(50 microL per 1.25 mL of reaction mixture) at 0.degree. C. The
reaction mixture is stirred at room temperature for 48-72 hours.
After TLC (EtOAc:EtOH:water, 3:1:1) indicates the completion of the
reaction, the volume of the solvents is reduced in vacuo. The
solution is applied to a column of silica for flash chromatography
(dichloromethane:methanol:aq. ammonia, 7:2:0.5) to furnish the
de-O-benzoylated products.
[0478] General Procedure L (GPL): Global Debenzylation.
[0479] Starting material is dissolved in THF and water (1/1, v/v, 3
mL for 10 mg) containing aqueous ammonia solution (150 .mu.L per 3
mL of reaction mixture) and treated with palladium hydroxide on
carbon (20% Pd, 5 times the weight of starting material). The
reaction mixture is stirred for 24-48 hours under hydrogen at
ambient temperature and pressure. After TLC (EtOAc:EtOH:water,
2:1:1) indicates the completion of the reaction the catalyst is
filtered off and washed with 50% aqueous THF. The solution is
concentrated to dryness and chromatography of the residue
(dichloromethane:methanol:aq. ammonia, 5:4:1) gives the final
products as ammonium salts. Some of the resulting materials are
dissolved in water, passed through a Dowex 50WX8-200 (Na.sup.+)
resin column (8.times.1 cm) and eluted with water. Fractions
containing the products are evaporated and dried in vacuo to
furnish sodium salts of final products.
[0480] Synthesis of 70
[0481] Compound 70 is prepared from compound 64 following general
procedure J: 135 mg, 81% yield, Rf=0.2
(dichloromethane:methanol:aq. ammonia, 7:2:0.5). HRMS (ESI) calcd
for C.sub.122H.sub.131N.sub.3O.sub.47S.sub.3Na (M+Na).sup.+ m/z
2508.7013, found 2508.65. .sup.1H-NMR (500 MHz, MeOD) 8.13-8.08 (m,
4H), 8.02 (d, J=7.3 Hz, 2H), 7.63 (q, J=7.0 Hz, 2H), 7.56 (t, J=7.6
Hz, 2H), 7.52 (q, J=9.8 Hz, 2H), 7.44 (t, J=8.0 Hz, 2H), 7.34-7.28
(m, 8 H), 7.27-7.23 (m, 8H), 7.22-7.17 (m, 8H), 7.16-7.13 (m, 6H),
7.12-7.07 (m, 6 H), 6.99 (d, J=9.1 Hz, 2H), 6.84 (d, J=9.1 Hz, 2H),
5.74 (d, J=3.1 Hz, 1H), 5.61 (d, J=3.9 Hz, 1H), 5.55 (d, J=3.9 Hz,
1H), 5.31 (t, J=3.7 Hz, 2H), 5.26-5.23 (m, 2H), 5.23-5.20 (m, 2H),
5.08 (d, J=3.3 Hz, 2H), 5.04 (d, J=3.4 Hz, 1H), 4.99 (d, J=3.3 Hz,
1H), 4.97 (d, J=3.1 Hz, 1H), 4.89-4.81 (m, 2H), 4.80-4.77 (m, 2H),
4.75 (d, J=3.5 Hz, 1H), 4.73-4.69 (m, 3H), 4.68 (d, J=11.6 Hz, 1H),
4.61 (d, J=11.1 Hz, 1H), 4.49 (t, J=7.5 Hz, 2H), 4.41-4.39 (m, 1H),
4.38-4.35 (m, 1H), 4.34-4.32 (m, 2H), 4.32-4.29 (m, 1H), 4.28-4.22
(m, 2H), 4.21-4.18 (m, 1H), 4.17-4.14 (m, 2H), 4.13 (d, J=3.9 Hz,
1H), 4.10-4.07 (m, 1H), 4.04 (t, J=9.5 Hz, 2H), 4.0 (t, J=9.5 Hz,
2H), 3.86-3.80 (m, 2H), 3.72 (s, 3H), 3.66 (t, J=7.2 Hz, 2H), 3.60
(d, J=9.2 Hz, 1H), 3.55-3.49 (m, 2H), 3.49-3.46 (m, 1H), 3.45 (s,
3H), 3.31 (s, 3H), 3.30 (s, 3H), 1.65 (s, 3H), 1.48 (s, 3H), 1.44
(s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta. 173.6, 173.3, 171.3,
171.1, 170.8, 167.4, 167.3, 157.0, 151.9, 140.0, 139.8, 138.99,
138.9, 138.8, 134.8, 134.7, 134.6, 131.2, 130.9, 130.8, 130.1,
130.0, 129.9, 129.6, 129.5, 129.4, 129.3, 129.2, 129.1, 129.0,
128.9, 128.8, 128.7, 128.5, 128.4, 128.1, 119.4, 115.7, 99.6, 99.4,
99.3, 98.4, 98.1, 97.3, 81.4, 79.8, 79.7, 79.2, 76.1, 75.8, 75.6,
75.3, 75.2, 74.9, 74.6, 74.2, 74.1, 74.0, 73.3, 72.9, 72.7, 72.2,
72.1, 71.9, 71.8, 71.7, 70.9, 70.6, 70.5, 67.3, 66.3, 56.1, 54.3,
53.8, 53.2, 53.0, 52.9, 22.9, 22.8, 227.
[0482] Synthesis of 71
[0483] Compound 71 is prepared from compound 65 following general
procedure J: 86 mg, 99%, R.sub.f=0.18
(dichloromethane/methanol/triethylamine 90:9:1). .sup.13C-NMR (125
MHz, CDCl.sub.3) .delta. 170.6, 170.5, 170.2, 168.5, 168.3, 167.9,
165.3, 165.2, 165.0, 155.7, 150.8, 139.1, 139.0, 138.6, 138.4,
137.1, 136.7, 133.6, 133.5, 130.0, 129.7, 129.2, 129.1, 129.0,
128.8, 128.6, 128.5, 128.4, 128.2, 128.1, 128.0, 127.8, 127.7,
127,6, 127.5, 127.4, 127.3, 118.7, 114.5, 100.7, 100.0, 99.8, 98.1,
97.8, 96.8, 81.8, 81.6, 81.4, 79.8, 77.8, 76.7, 76.6, 75.8, 75.2,
75.1, 74.85, 74.8, 74.7, 74.5, 74.3, 74.2, 74.0, 73.9, 73.7, 73.6,
70.8, 70.5, 70.0, 65.6, 64.2, 64.0, 55.6, 52.7, 52.4, 52.1, 52.0,
51.9, 46.5, 23.0, 22.8, 22.7, 8.9
[0484] Synthesis of 72
[0485] Compound 72 is prepared from compound 66 following general
procedure J: 13 mg, 78% yield, Rf=0.2 (dichloromethane:methanol:aq.
ammonia, 7:2:0.5). HRMS (ESI) calcd for
C.sub.158H.sub.170N.sub.4O.sub.62S.sub.4Na (M+Na).sup.+ m/z
3265.9053, found 3265.8494. .sup.1H-NMR (500 MHz, MeOD) 8.13-8.08
(m, 6H), 8.01 (d, J=7.5 Hz, 2H), 7.63-7.59 (m, 2H), 7.59-7.54 (m,
2H), 7.53-7.47 (m, 2H), 7.46-7.41 (m, 6H), 7.33-7.28 (m, 10 H),
7.28-7.22 (m, 15H), 7.22-7.18 (m, 10H), 7.17-7.14 (m, 6H), 7.13 (d,
J=7.4 Hz, 2H), 7.09 (d, J=7.2 Hz, 2H), 6.99 (d, J=9.1 Hz, 2H), 6.84
(d, J=8.9 Hz, 2H), 5.73 (d, J=3.4 Hz, 1H), 5.59 (d, J=3.9 Hz, 1H),
5.56-5.52 (m, 2H), 5.31-5.28 (m, 2H), 5.25-5.17 (m, 3H), 5.07 (d,
J=3.6 Hz, 1H), 5.03 (t, J=3.9 Hz, 2H), 4.99 (d, J=3.9 Hz, 1H),
4.97-4.96 (m, 2H), 4.88 (d, J=3.0 Hz, 1H), 4.81-4.76 (m, 3H),
4.74-4.72 (m, 2H), 4.72-4.70 (m, 2H), 4.70-4.68 (m, 3H), 4.68-4.65
(m, 2H), 4.63 (d, J=11.5 Hz, 1H), 4.50 (d, J=10.4 Hz, 2H),
4.39-4.32 (m, 6H), 4.32-4.28 (m, 4H), 4.27-4.22 (m, 4H), 4.21-4.17
(m, 2H), 4.17-4.11 (m, 3H), 4.10 (t, J=3.8 Hz, 2H), 4.08 (t, J=3.8
Hz, 2H), 4.03-3.98 (m, 3H), 3.98-3.94 (m, 2H), 3.84-3.79 (m, 2H),
3.73 (s, 3H), 3.63-3.57 (m, 2H), 3.56-3.50 (m, 2H), 3.49-3.47 (m,
2H), 3.47 (s, 3H), 3.30 (s, 3H), 3.28 (s, 3H), 3.27 (s, 3H), 1.65
(s, 3H), 1.49 (s, 3H), 1.47 (s, 3H), 1.44 (s, 3H); .sup.13C-NMR
(125 MHz, MeOD) .delta. 173.6, 173.3, 171.2, 170.8, 167.3, 157.0,
152.0, 140.0, 139.8, 138.9, 138.88, 138.83, 138.7, 134.8, 134.6,
131.2, 130.8, 130.1, 130.0, 129.9, 129.5, 129.3, 129.2, 129.1,
128.9, 128.8, 128.7, 128.5, 128.4, 128.2, 128.0, 119.5, 115.8,
99.66, 99.4, 99.3, 99.1, 98.3, 98.0, 97.2, 81.4, 79.8, 79.1, 76.1,
75.8, 75.6, 75.5, 74.2, 74.9, 74.6, 74.5, 74.2, 74.1, 73.3, 72.9,
72.6, 72.5, 72.1, 71.8, 70.8, 70.7, 70.5, 67.2, 66.3, 56.1, 54.3,
53.8, 53.2, 53.0, 22.9, 22.8, 22.7.
[0486] Synthesis of 73
[0487] Compound 73 is prepared from compound 67 following general
procedure J: 46 mg, quant., R.sub.f=0.15
(dichloromethane/methanol/triethylamine 90:9:1). .sup.13C-NMR (125
MHz, CDCl.sub.3) .delta. 170.5, 170.4, 170.1, 168.4, 168.35, 168.3,
167.9, 165.3, 165.2, 165.0, 155.7, 150.8, 139.1, 139.0, 138.6,
138.4, 137.1, 136.8, 133.6, 133.5, 130.1, 129.7, 129.2, 129.1,
129.0, 128.8, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 127.8,
127.7, 127.6, 127.5, 127.4, 118.7, 114.5, 100.7, 100.1, 99.9, 98.3,
97.8, 96.8, 81.7, 81.4, 79.7, 77.8, 76.7, 76.6, 76.0, 75.2, 75.1,
74.9, 74.8, 74.7, 74.6, 74.4, 74.3, 74.1, 73.9, 73.8, 73.7, 73.6,
70.9, 70.6, 70.1, 65.5, 64.1, 64.0, 63.9, 55.6, 52.7, 52.6, 52.4,
52.2, 52.1, 52.0, 51.95, 51.9, 46.3, 23.1, 22.8, 22.7, 9.8
[0488] Synthesis of 74
[0489] Compound 74 is prepared from compound 68 following general
procedure J: 25 mg, 94% yield, Rf=0.15
(dichloromethane:methanol:aq. ammonia, 7:2:0.5). HRMS (ESI) calcd
for C.sub.194H.sub.209N.sub.5O.sub.77S.sub.5Na.sub.3 (M+3Na).sup.3+
m/z 4069.09, found 4070.2. .sup.1H-NMR (500 MHz, MeOD) 8.13-8.08
(m, 8H), 8.01 (d, J=8.9 Hz, 2H), 7.64-7.59 (m, 6H), 7.59-7.54 (m,
6H), 7.53-7.45 (m, 4H), 7.44-7.39 (m, 6H), 7.32-7.28 (m, 12 H),
7.28-7.21 (m, 14H), 7.21-7.19 (m, 10H), 7.18-7.13 (m, 8H),
7.12-7.06 (m, 4H), 6.99 (d, J=9.5 Hz, 2H), 6.84 (d, J=9.5 Hz, 2H),
5.73 (d, J=3.4 Hz, 1H), 5.59 (d, J=4.0 Hz, 1H), 5.56-5.52 (m, 2H),
5.31-5.28 (m, 2H), 5.24-5.20 (m, 3H), 5.19-5.16 (m, 3H), 5.08 (d,
J=4.2 Hz, 1H), 5.03-5.0 (m, 2H), 4.99-4.96 (m, 2H), 4.87 (d, J=2.9
Hz, 1H), 4.81-4.76 (m, 3H), 4.75-4.70 (m, 4H), 4.70-4.64 (m, 5H),
4.62 (d, J=11.2 Hz, 1H), 4.49 (d, J=11.2 Hz, 2H), 4.39-4.34 (m,
6H), 4.33-4.27 (m, 8H), 4.26-4.21 (m, 6H), 4.20-4.17 (m, 8H),
4.17-4.12 (m, 8H), 4.11-4.04 (m, 2H), 4.03-3.96 (m, 3H), 3.84-3.80
(m, 2H), 3.74 (s, 3H), 3.64-3.61 (m, 2H), 3.61-3.56 (m, 2H),
3.55-3.50 (m, 1H), 3.48-3.43 (m, 2H), 3.47 (s, 3H), 3.31 (s, 3H),
3.30 (s, 3H), 3.28 (s, 3H), 3.26 (s, 3H), 1.65 (s, 3H), 1.49 (s,
3H), 1.48 (s, 3H), 1.47 (s, 3H), 1.43 (s, 3H); .sup.13C-NMR (MeOD)
.delta. 173.5, 170.8, 167.3, 158.1, 153.2, 139.9, 138.8, 134.7,
131.2, 130.9, 129.9, 129.5, 129.3, 129.1, 128.9, 128.8, 128.5,
128.4, 128.2, 128.0, 119.5, 115.8, 99.7, 99.5, 99.3, 99.2, 98.5,
98.3, 97.2, 97.1, 79.9, 76.1, 75.5, 75.1, 74.5, 74.1, 71.9, 66.3,
56.1, 53.8, 52.9, 49.6, 22.8.
[0490] Synthesis of 143
[0491] Compound 143 is prepared from compound 142 following general
procedure J: 132 mg, 27.7 .mu.mol, 83% yield, R.sub.f=0.15
(dichloromethane:methanol:aq. ammonia, 7:2:0.5); HRMS (ESI) calcd
for C.sub.230H.sub.248N.sub.6O.sub.92S.sub.6Na.sub.3 (M+3Na).sup.+3
m/z 1608.0852, found 1608.0801. .sup.1H-NMR (500 MHz, MeOD)
8.15-8.08 (m, 8H), 8.01 (d, J=8.6 Hz, 2H), 7.65-7.57 (m, 5H),
7.55-7.44 (m, 8H), 7.44-7.40 (m, 6H), 7.35-7.28 (m, 24 H),
7.27-7.21 (m, 18H), 7.20-7.14 (m, 16H), 7.14-7.05 (m, 8H), 6.99 (d,
J=9.0 Hz, 2H), 6.84 (d, J=9.0 Hz, 2H), 5.74 (d, J=4.3 Hz, 1H), 5.61
(d, J=3.0 Hz, 1H), 5.58-5.53 (m, 3H), 5.32 (t, J=3.8 Hz, 2H),
5.26-5.21 (m, 2H), 5.21-5.16 (m, 2H), 5.06 (d, J=3.4 Hz, 1H),
5.03-4.99 (m, 2H), 4.99 (d, J=3.4 Hz, 1H), 4.96 (d, J=2.4 Hz, 1H),
4.88 (d, J=2.4 Hz, 1H), 4.82-4.73 (m, 3H), 4.75-4.71 (m, 4H),
4.70-4.65 (m, 5H), 4.62 (d, J=11.0 Hz, 1H), 4.52-4.44 (m, 8H),
4.39-4.35 (m, 10H), 4.35-4.27 (m, 12H), 4.26-4.18 (m, 6H),
4.18-4.11 (m, 12H), 4.11-4.04 (m, 4H), 4.04-3.95 (m, 4H), 3.87-3.81
(m, 2H), 3.73 (s, 3H), 3.71 (s, 3H), 3.69 (s, 6H), 3.64-3.57 (m,
4H), 3.56-3.51 (m, 2H), 3.31 (s, 3H), 3.30 (s, 6H), 3.29 (d, J=4.5
Hz, 1H), 1.67 (s, 3H), 1.51 (s, 3H), 1.50 (s, 6H), 1.49 (s, 3H),
1.46 (s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta. 173.5, 173.3,
171.2, 170.9, 167.5, 167.4, 167.3, 164.9, 157.1, 152.0, 140.0,
139.9, 139.0, 138.9, 138.8, 138.7, 134.9, 134.7, 131.2, 130.9,
130.8, 130.0, 129.6, 129.3, 129.2, 129.17, 129.12, 129.0, 128.9,
128.7, 128.6, 128.4, 128.2, 119.5, 115.8, 99.7, 99.3, 98.5, 98.4,
98.2, 97.4, 81.4, 79.8, 79.2, 76.2, 76.1, 76.0, 75.6, 75.2, 74.6,
74.2, 74.0, 73.5, 73.1, 72.7, 72.6, 72.2, 72.1, 71.9, 70.9, 70.7,
70.6, 67.3, 66.3, 56.2, 55.2, 54.3, 53.8, 53.2, 53.1, 52.9, 48.6,
22.9, 22.8, 22.7.
[0492] Synthesis of 145
[0493] Compound 145 is prepared from compound 80 following general
procedure J: 69 mg, 18.1 .mu.mol, 98% yield, R.sub.f=0.15
(dichloromethane:methanol:aq. ammonia, 7:2:0.5); HRMS (ESI) calcd
for C.sub.154H.sub.178N.sub.5O.sub.87S.sub.10NH.sub.4 (M-4H).sup.-4
m/z 956.1735, found 956.1740. .sup.1H-NMR (500 MHz, MeOD) 7.45-7.42
(m, 4H), 7.35-7.31 (m, 6H), 7.30-7.27 (m, 18H), 7.26-7.22 (m, 12
H), 7.22-7.19 (m, 10H), 7.19-7.15 (m, 5H), 7.07 (d, J=9.0 Hz, 2H),
6.85 (d, J=9.1 Hz, 2H), 5.88 (d, J=3.0 Hz, 1H), 5.60-5.56 (m, 2H),
5.02-4.93 (m, 5H), 4.92-4.74 (m, 12H), 4.74-4.70 (m, 3H), 4.70-4.61
(m, 6H), 4.59-4.49 (m, 5H), 4.39-4.33 (m, 8H), 4.33-4.23 (m, 10H),
4.23 (d, J=9.5 Hz, 1H), 4.19-4.16 (m, 6H), 4.16-4.13 (m, 6H), 4.13
(d, J=3.5 Hz, 1H), 4.08-4.03 (m, 4H), 4.02-3.99 (m, 2H), 3.98-3.96
(m, 1H), 3.96-3.92 (m, 2H), 3.90-3.82 (m, 3H), 3.81-3.78 (m, 2H),
3.73 (s, 3H), 3.66-3.61 (m, 2H), 3.31 (s, 3H), 3.309 (s, 3H), 3.306
(s, 3H), 3.303 (s, 3H), 3.29 (s, 3H), 1.96 (s, 3H), 1.93 (s, 6H),
1.92 (s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta. 175.9, 174.1,
159.2, 156.6, 152.1, 140.3, 139.7, 139.2, 138.7, 129.7, 129.6,
129.5, 129.4, 129.3, 129.1, 128.9, 128.8, 128.7, 128.6, 128.2,
119.4, 115.7, 99.7, 99.6, 99.5, 99.4, 99.2, 98.9, 81.8, 79.3, 76.3,
75.3, 74.0, 72.8, 72.7, 72.6, 71.5, 69.3, 67.9, 67.2, 56.1, 54.5,
49.5, 41.0, 23.3.
[0494] Synthesis of 146
[0495] Compound 146 is prepared from compound 144 following general
procedure J: 69 mg, 18.1 .mu.mol, 98% yield, R.sub.f=0.15
(dichloromethane:methanol:aq. ammonia, 7:2:0.5); HRMS (ESI) calcd
for C.sub.182H.sub.212N.sub.6O.sub.104S.sub.12Na (M+Na).sup.+ m/z
4553.81, found 4553.01. .sup.1H-NMR (500 MHz, MeOD) 7.46-7.41 (m,
4H), 7.35-7.31 (m, 6H), 7.30-7.27 (m, 24H), 7.26-7.23 (m, 16 H),
7.22-7.18 (m, 10H), 7.18-7.13 (m, 5H), 7.07 (d, J=8.9 Hz, 2H), 6.84
(d, J=9.0 Hz, 2H), 5.88 (d, J=3.0 Hz, 1H), 5.61-5.55 (m, 2H),
5.04-4.94 (m, 5H), 4.93-4.74 (m, 12H), 4.74-4.69 (m, 3H), 4.69-4.60
(m, 6H), 4.59-4.46 (m, 5H), 4.41-4.35 (m, 10H), 4.33-4.26 (m, 10H),
4.26 (d, J=11.0 Hz, 1H), 4.2-4.11 (m, 8H), 4.08-4.03 (m, 5H), 4.03
(d, J=9.8 Hz, 1H), 3.98-3.96 (m, 5H), 3.96-3.92 (m, 5H), 3.91-3.85
(m, 3H), 3.83-3.80 (m, 2H), 3.80-3.77 (m, 2H), 3.76-3.73 (m, 1H),
3.73 (s, 3H), 3.66-3.61 (m, 2H), 3.313 (s, 3H), 3.31 (s, 3H), 3.306
(s, 6H), 3.303 (s, 3H), 3.3 (s, 3H), 1.97 (s, 6H), 1.95 (s, 3H),
1.94 (s, 6H), 1.92 (s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta.
176.0, 175.5, 174.1, 159.2, 156.6, 152.1, 140.3, 139.7, 139.1,
138.7, 129.7, 129.5, 129.3, 129.2, 129.0, 128.9, 128.8, 128.7,
128.6, 128.5, 128.2, 119.4, 115.7, 99.7, 99.6, 99.5, 99.4, 98.9,
98.2, 96.7, 81.8, 79.3, 76.3, 75.3, 74.0, 72.7, 72.6, 72.5, 72.2,
71.5, 70.0, 69.3, 68.0, 67.2, 56.1, 54.5, 49.5, 41.0, 23.3.
[0496] Synthesis of 75
[0497] Compound 75 is prepared from compound 69 following general
procedure J: 130 mg, quant., R.sub.f=0.15
(dichloromethane/methanol/triethylamine 90:9:1); MS (ESI, negative
mode) calcd for C.sub.194H.sub.209N.sub.5O.sub.77S.sub.5
(M-5HNEt.sub.3+2H).sup.2- m/z (%): 1999.05 (30), 1999.55 (70),
2000.05 (100), 2000.55 (90), 2001.06 (65), 2001.56 (25) found
1999.05 (30), 1999.55 (70), 2000.05 (90), 2000.55 (80), 2001.05
(65), 2001.55 (35). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
170.35, 170.3, 168.5, 168.3, 168.25, 168.2, 167.9, 165.2, 165.1,
155.6, 150.9, 139.1, 138.6, 138.4, 137.0, 133.4, 130.0, 129.7,
129.3, 129.1, 129.0, 128.8, 128.5, 128.4, 128.3, 128.2, 128.0,
127.9, 127.8, 127.7, 127.6, 127.5, 127.3, 127.2, 118.6, 114.4,
100.4, 100.0, 99.9, 98.3, 97.5, 96.8, 81.9, 81.4, 79.8, 77.7, 76.4,
76.0, 75.0, 74.9, 74.8, 74.6, 74.5, 74.4, 74.1, 74.0, 73.7, 73.5,
70.9, 70.4, 70.0, 65.4, 63.9, 55.6, 54.5, 52.6, 52.1, 51.85, 51.8,
46.3, 23.1, 22.8, 8.6
[0498] Synthesis of 76
[0499] Compound 76 is prepared from compound 70 following general
procedure K: 52 mg, 81% yield, Rf=0.2 (EtOAc:EtOH:water, 3:1:1).
HRMS (ESI) calcd for
C.sub.98H.sub.113N.sub.3O.sub.44S.sub.3Na.sub.2 (M+2Na).sup.2+ m/z
2177.5654, found 2177.62. .sup.1H-NMR (500 MHz, MeOD) 7.44-7.40 (m,
3H), 7.39-7.35 (m, 5 H), 7.34-7.31 (m, 5H), 7.29-7.26 (m, 5H),
7.26-7.21 (m, 8H), 7.20-7.17 (m, 6H), 7.15-7.13 (m, 3 H), 7.09 (d,
J=8.9 Hz, 2H), 6.85 (d, J=8.9 Hz, 2H), 5.49 (d, J=4.0 Hz, 1H), 5.30
(d, J=4.6 Hz, 1H), 5.25 (d, J=4.3 Hz, 1H), 4.87-4.77 (m, 6H),
4.76-4.74 (m, 2H), 4.73-4.68 (m, 2H), 4.67-4.65 (m, 2H), 4.65-4.61
(m, 3H), 4.61 (d, J=4.6 Hz, 1H), 4.46 (d, J=10.7 Hz, 1H), 4.40 (d,
J=11.5 Hz, 1H), 4.34-4.29 (m, 6H), 4.26-4.17 (m, 8H), 4.13-4.04 (m,
5H), 4.0-3.95 (m, 3H), 3.94-3.84 (m, 2H), 3.82-3.77 (m, 2H), 3.73
(s, 3H), 3.71-3.65 (m, 2H), 3.65-3.59 (m, 2H), 1.79 (s, 3H), 1.69
(s, 3H), 1.66 (s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta. 173.2,
156.5, 152.4, 139.9, 139.8, 139.5, 139.1, 129.6, 129.5, 129.3,
129.0, 128.8, 128.75, 128.7, 128.6, 119.2, 115.7, 101.7, 100.9,
100.5, 99.2, 98.1, 82.2, 79.1, 76.2, 76.1, 74.9, 73.0, 71.6, 68.6,
67.6, 67.3, 56.1, 54.3, 49.5, 22.9.
[0500] Synthesis of 77
[0501] Compound 77 is prepared from compound 71 following general
procedure K: 29 mg, 72%, R.sub.f=0.15 (EtOAc/ethanol/water 3:1:1).
.sup.13C-NMR (125 MHz, methanol-D4, HSQC) .delta. 129.6, 129.3,
129.2, 129.1, 128.5, 119.4, 115.5, 104.1, 103.8, 98.1, 86.3,82.1,
80.9, 79.4, 78.6, 77.9, 76.4, 76.3, 75.9, 75.8, 75.6, 75.3, 75.2,
75.1, 74.9, 74.8, 71.2, 67.4, 67.0, 56.1, 54.6, 53.9, 47.4, 23.0,
9.7
[0502] Synthesis of 78
[0503] Compound 78 is prepared from compound 72 following general
procedure K: 29 mg, 85% yield, R.sub.f=0.22 (EtOAc:EtOH:water,
3:1:1). HRMS (ESI) calcd for
C.sub.126H.sub.146N.sub.4O.sub.58S.sub.4Na (M+Na).sup.+ m/z
2793.7379, found 2793.7398. .sup.1H-NMR (500 MHz, MeOD) 7.43-7.40
(m, 4H), 7.40-7.36 (m, 5 H), 7.35-7.31 (m, 8H), 7.30-7.25 (m, 10H),
7.25-7.22 (m, 8H), 7.21-7.15 (m, 6H), 7.15-7.11 (m, 4 H), 7.09 (d,
J=9.0 Hz, 2H), 6.85 (d, J=9.0 Hz, 2H), 5.48 (d, J=3.4 Hz, 1H), 5.27
(d, J=3.9 Hz, 1H), 5.24 (d, J=4.5 Hz, 1H), 5.21 (d, J=4.5 Hz, 1H),
4.87-4.85 (m, 3H), 4.84-4.77 (m, 4H), 4.77-4.74 (m, 2H), 4.74-4.70
(m, 2H), 4.70-4.68 (m, 2H), 4.66-4.60 (m, 3H), 4.45-4.43 (m, 1H),
4.43-4.41 (m, 1H), 4.40-4.38 (m, 1H), 4.38-4.34 (m, 5H), 4.34-4.32
(m, 3H), 4.32-4.30 (m, 4H), 4.29-4.27 (m, 2H), 4.27-4.25 (m, 3H),
4.25-4.20 (m, 4H), 4.19 (d, J=3.7 Hz, 1H), 4.17 (d, J=3.1 Hz, 1H),
4.13-4.05 (m, 4H), 4.04 (t, J=4.1 Hz, 2H), 4.01 (d, J=4.1 Hz, 1H),
3.97-3.93 (m, 3H), 3.92-3.83 (m, 4H), 3.82-3.76 (m, 2H), 3.74 (s,
3H), 3.73-3.68 (m, 2H), 3.67-3.60 (m, 2H), 1.80 (s, 3H), 1.66 (s,
3H), 1.63 (s, 3H), 1.62 (s, 3H); .sup.13C-NMR (125 MHz, MeOD)
.delta. 176.5, 175.7, 175.4, 156.7, 155.8, 154.1, 139.8, 139.1,
129.7, 129.6, 129.5, 129.4, 129.3, 129.2, 129.1, 128.9, 128.8,
128.7, 128.5, 119.3, 115.7, 102.4, 102.3, 101.8, 101.7, 101.6,
100.1, 97.9, 97.8, 87.3, 86.5, 81.7, 81.1, 76.6, 76.2, 76.1, 73.1,
72.9, 71.6, 69.5, 68.7, 67.4, 56.1, 54.4, 53.9, 53.5, 22.9.
[0504] Synthesis of 79
[0505] Compound 79 is prepared from compound 73 following general
procedure K: 29 mg, 94%, R.sub.f=0.6 (EtOAc/ethanol/water 2:2:1).
.sup.13C-NMR (125 MHz, methanol-D4) .delta. 176.6, 175.8, 175.3,
173.1, 173.0, 155.9, 153.2, 140.3, 139.9, 139.6, 129.9, 129.5,
129.4, 129.2, 129.1, 128.6, 128.5, 119.6, 115.5, 104.1, 104.0,
103.9, 98.6, 98.2, 98.0, 86.4, 86.1, 85.8, 81.8, 80.5, 79.3, 78.3,
78.2, 77.6, 76.6, 76.2, 75.9, 75.8, 75.7, 75.3, 75.1, 74.9, 74.8,
74.6, 71.3, 71.2, 71.1, 67.4, 67.1, 56.1, 54.4, 53.5, 53.4, 53.3,
23.0
[0506] Synthesis of 80
[0507] Compound 80 is prepared from compound 74 following general
procedure K: 14 mg, 82% yield, R.sub.f=0.21 (EtOAc:EtOH:water,
3:1:1). .sup.1H-NMR (500 MHz, MeOD) 7.43-7.40 (m, 5H), 7.39-7.35
(m, 6 H), 7.35-7.30 (m, 12H), 7.29-7.23 (m, 16H), 7.22-7.15 (m,
12H), 7.15-7.13 (m, 4 H), 7.09 (d, J=9.0 Hz, 2H), 6.85 (d, J=9.2
Hz, 2H), 5.47 (d, J=4.0 Hz, 1H), 5.27 (d, J=3.9 Hz, 1H), 5.24-19
(m, 3H), 4.94-4.75 (m, 8H), 4.74-4.72 (m, 6H), 4.72-4.69 (m, 2H),
4.69-4.67 (m, 4H), 4.66-4.60 (m, 3H), 4.46-4.42 (m, 8H), 4.41-4.38
(m, 10H), 4.37-4.34 (m, 4H), 4.33-4.28 (m, 5H), 4.27-4.23 (m, 6H),
4.23-4.17 (m, 8H), 4.14-4.06 (m, 4H), 4.04-3.97 (m, 3H), 3.93-3.82
(m, 4H), 3.80-3.76 (m, 3H), 3.74 (s, 3H), 3.73-3.66 (m, 2H),
3.66-3.63 (m, 2H), 1.81 (s, 3H), 1.65 (s, 3H), 1.62 (s, 6H), 1.61
(s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta. 8 173.2, 156.5,
152.5, 139.9, 139.5, 139.2, 129.7, 129.6, 129.4, 129.1, 129.0,
128.9, 128.9, 128.8, 128.7, 128.5, 119.2, 115.7, 101.7, 100.8,
99.9, 99.8, 79.2, 76.1, 72.9, 71.6, 70.5, 69.2, 56.1, 55.1, 54.4,
49.6, 30.9, 30.7, 22.9.
[0508] Synthesis of 144
[0509] Compound 144 is prepared from compound 143 following general
procedure K: 85 mg, 20.98 .mu.mol, 76% yield, R.sub.f=0.2
(EtOAc:EtOH:water, 3:1:1); HRMS (ESI) calcd for
C.sub.182H.sub.210N.sub.6O.sub.86S.sub.6Na.sub.2 (M+2Na).sup.+2 m/z
2046.5131, found 2046.5109. .sup.1H-NMR (500 MHz, MeOD) 7.43-7.38
(m, 4H), 7.36-7.32 (m, 6 H), 7.32-7.29 (m, 24H), 7.30-7.25 (m,
20H), 7.25-7.23 (m, 6H), 7.23-7.18 (m, 5 H), 7.09 (d, J=8.9 Hz,
2H), 6.84 (d, J=8.9 Hz, 2H), 5.49-5.47 (m, 2H), 5.40 (d, J=5.3 Hz,
1H), 5.34-5.32 (m, 2H), 5.28-5.25 (m, 4H), 5.24-5.21 (m, 3H),
5.03-4.79 (m, 12H), 4.79-4.75 (m, 6H), 4.75-4.73 (m, 4H), 4.73-4.67
(m, 6H), 4.66-4.59 (m, 7H), 4.66-4.60 (m, 3H), 4.49-4.41 (m, 8H),
4.39-4.31 (m, 8H), 4.31-4.24 (m, 5H), 4.24-4.19 (m, 6H), 4.17-4.12
(m, 3H), 4.11-4.05 (m, 5H), 4.05-3.98 (m, 3H), 3.94-3.81 (m, 4H),
3.79-3.74 (m, 3H), 3.73 (s, 3H), 3.66-3.62 (m, 3H), 1.82 (s, 3H),
1.66 (s, 3H), 1.628 (s, 6H), 1.621 (s, 3H), 1.61 (s, 3H);
.sup.13C-NMR (125 MHz, MeOD) .delta. 175.8, 173.3, 156.5, 152.4,
140.2, 140.1, 139.9, 139.7, 139.5, 139.2, 139.1, 130.5, 129.9,
129.7, 129.6, 129.5, 129.3, 129.2, 129.1, 128.9, 128.8, 128.7,
128.6, 101.7, 98.1, 101.8, 101.7, 99.7, 99.6, 98.4, 98.3, 98.1,
82.2, 80.8, 79.2, 76.9, 76.5, 76.3, 75.9, 75.3, 74.9, 73.0, 72.8,
72.3, 71.5, 70.7, 70.3, 69.8, 68.4, 67.6, 56.2, 55.2, 54.3, 49.9,
39.3, 22.9.
[0510] Synthesis of 81
[0511] Compound 81 is prepared from compound 75 following general
procedure K: 75 mg, 75%, R.sub.f=0.05 (EtOAc/ethanol/water 3:1:1).
MS (ESI, negative mode) calcd for
C.sub.154H.sub.169N.sub.5O.sub.72S.sub.5 (M-10HNEt.sub.3+8H).sup.2-
m/z (%): 1703.95 found 1703.93. .sup.13C-NMR (125 MHz, methanol-D4)
.delta. 175.9, 175.2, 174.4, 173.1, 172.9, 156.9, 153.2, 140.4,
140.3, 139.9, 139.6, 129.5, 129.4, 139.35, 129.3, 129.2, 129.0,
128.6, 128.5, 119.6, 115.5, 104.1, 104.05, 104.0, 103.9, 98.7,
98.2, 86.4, 86.1, 85.8, 81.9, 80.4, 80.2, 79.3, 78.2, 77.9, 77.7,
76.6, 76.5, 76.3, 75.9, 75.8, 75.7, 75.6, 75.3, 75.2, 75.1, 74.7,
71.4, 71.1, 67.3, 67.0, 56.1, 54.3, 53.4, 22.9
[0512] Synthesis of 82
[0513] Compound 82 is prepared from compound 78 following general
procedure J: 24 mg, 90% yield, Rf=0.15
(dichloromethane:methanol:aq. ammonia, 7:2:0.5). .sup.1H-NMR (500
MHz, MeOD) 7.47-7.42 (m, 4H), 7.40-7.35 (m, 6H), 7.35-7.31 (m, 8
H), 7.30-7.25 (m, 6H), 7.25-7.20 (m, 7H), 7.20-7.15 (m, 8H),
7.14-7.11 (m, 6H), 7.08 (d, J=8.3 Hz, 2H), 6.85 (d, J=9.0 Hz, 2H),
5.89-5.86 (m, 1H), 5.62-5.57 (m, 1H), 5.55 (d, J=9.8 Hz, 1H), 5.49
(d, J=6.9 Hz, 1H), 5.44-5.40 (m, 4H), 5.39-5.36 (m, 3H), 5.35-5.32
(m, 4H), 5.29-5.27 (m, 3H), 5.26 (t, J=9.8 Hz, 2H),), 4.99-4.75 (m,
3H), 4.75-4.72 (m, 4H), 4.71-4.67 (m, 2H), 4.67-4.59 (5H),
4.59-4.50 (m, 8H), 4.38-4.28 (m, 6H), 4.27-4.13 (m, 4H), 4.12-4.01
(m, 6H), 4.0-3.81 (m, 2H), 3.80-3.76 (m, 2H), 3.74 (s, 3H),
3.72-3.67 (m, 2H), 3.67-3.60 (m, 2H), 1.62 (s, 3H), 1.61 (s, 3H),
1.49 (s, 3H), 1.48 (s, 3H); .sup.13C-NMR (125 MHz, MeOD) .delta.
174.1, 173.3, 156.6, 140.2, 139.8, 139.1, 139.0, 138.6, 129.7,
129.5, 129.3, 129.28, 129.2, 129.1, 129.0, 128.9, 128.8, 128.3,
128.2, 119.4, 115.7, 99.5, 99.2, 99.1, 99.0, 98.7, 98.5, 76.2,
76.1, 73.7, 73.1, 71.6, 68.9, 67.4, 56.1, 54.4, 53.8, 23.3,
22.9.
[0514] Synthesis of 83
[0515] Compound 83 is prepared from compound 79 following general
procedure J: 15 mg, quant. .sup.13C-NMR (125 MHz, methanol-D4,
HSQC) .delta. 130.0, 129.9, 129.5, 129.4, 129.2, 128.8, 120.1,
115.5, 102.4, 101.6, 101.5, 98.6, 97.5, 97.4, 83.9, 81.9, 81.5,
81.1, 81.0, 80.4, 79.8, 79.6, 79.3, 78.2, 76.6, 76.5, 76.1, 76.0,
75.9, 75.6, 75.5, 75.0, 74.9, 74.8, 74.5, 74.4, 74.3, 73.9, 71.6,
71.4, 71.3, 67.4, 67.0, 66.8, 56.2, 53.4, 52.9, 22.1
[0516] Synthesis of 84
[0517] Compound 84 is prepared from compound 76 following general
procedure L: 9 mg, 85% yield, Rf=0.2 (EtOAc:EtOH:water, 2:1:1).
HRMS (ESI) calcd for C.sub.49H.sub.71N.sub.3O.sub.44S.sub.3Na
(M+Na).sup.+ m/z 1547.2368, found 1547.26. .sup.1H-NMR (500 MHz,
D.sub.2O) 7.13 (d, J=9.2 Hz, 2H), 6.98 (d, J=9.1 Hz, 2H), 5.45 (d,
J=3.2 Hz, 1H), 5.16 (d, J=3.8 Hz, 1H), 5.14 (d, J=3.8 Hz, 1H), 4.98
(d, J=3.2 Hz, 1H), 4.89-4.86 (m, 1H), 4.75 (d, J=3.2 Hz, 1H), 4.33
(d, J=3.0 Hz, 1H), 4.30 (d, J=3.0 Hz, 1H), 4.23 (d, J=13.0 Hz, 1H),
4.17-4.15 (m, 4H), 4.11 (t, J=3.5 Hz, 2H), 4.08-4.04 (m, 5H),
4.04-4.02 (m, 2H), 4.01-3.92 (m, 5H), 3.93 (d, J=3.5 Hz, 1H),
3.91-3.88 (m, 2H), 3.88-3.84 (m, 3H), 3.79 (s, 3H), 3.75-3.70 (m,
4H), 3.69-3.67 (m, 2H), 3.67-3.65 (m, 2H), 3.65-3.63 (m, 2H),
3.56-3.55 (m, 1H), 3.54-3.53 (m, 1H), 3.52-3.51 (m, 1H), 1.99 (s,
6H), 1.98 (s, 3H); .sup.13C-NMR (125 MHz, D.sub.2O) .delta. 174.5,
174.4, 1742, 154.8, 150.3, 119.3, 115.2, 101.9, 101.8, 100.7,
100.6, 94.9, 77.1, 76.8, 74.2, 74.1, 73.9, 71.1, 70.4, 69.9, 69.4,
69.2, 69.18, 69.1, 68.9, 68.8, 68.5, 68.4, 67.9, 66.5, 66.2, 55.9,
53.6, 53.4, 48.9, 22.0.
[0518] Synthesis of 85
[0519] Compound 85 is prepared from compound 77 following general
procedure L: 4 mg, 92%, R.sub.f=0.1 (EtOAc/ethanol/water 2:2:1).
.sup.13C-NMR (125 MHz, D.sub.2O) .delta. 175.0, 174.9, 174.7,
174.5, 174.3, 154.8, 151.0, 118.4, 115.1, 102.1, 102.0, 101.3,
97.0, 96.7, 77.7, 77.3, 76.8, 76.5, 76.4, 76.3, 76.1, 76.0, 73.6,
73.5, 73.4, 70.7, 70.1, 69.1, 69.0, 68.8, 68.5, 67.9, 66.4, 65.8,
55.9, 53.6, 53.1, 22.0
[0520] Synthesis of 86
[0521] Compound 86 is prepared from compound 78 following general
procedure L: 4 mg, 94% yield, Rf=0.2 (EtOAc:EtOH:water, 2:1:1).
HRMS (ESI) calcd for C.sub.63H.sub.92N.sub.4O.sub.58S.sub.4Na.sub.3
(M+3Na).sup.3+ m/z 2029.2948, found 2029.2834. .sup.1H-NMR (500
MHz, D.sub.2O) 7.13 (d, J=9.2 Hz, 2H), 6.98 (d, J=9.2 Hz, 2H), 5.47
(d, J=3.4 Hz, 1H), 5.17 (d, J=4.2 Hz, 1H), 5.14 (d, J=3.4 Hz, 1H),
4.99-4.96 (m, 2H), 4.96 (d, J=3.41 Hz, 1H), 4.81 (d, J=2.3 Hz, 1H),
4.77 (d, J=3.0 Hz, 1H), 4.73-4.65 (m, 3H), 4.33-4.30 (m, 3H), 4.30
(d, J=3.7 Hz, 1H), 4.22-4.20 (m, 2H), 4.19-4.17 (m, 4H), 4.17-4.15
(m, 2H), 4.12 (t, J=3.0 Hz, 2H), 4.08-4.04 (m, 5H), 4.04-4.02 (m,
4H), 4.0-3.94 (m, 5H), 3.94-3.89 (m, 4H), 3.89-3.86 (m, 2H),
3.83-3.81 (m, 3H), 3.79 (s, 3H), 3.75-3.69 (m, 4H), 3.68-3.67 (m,
2H), 3.66-3.65 (m, 2H), 3.64-3.63 (m, 2H), 3.57-3.56 (m, 1H),
3.55-3.54 (m, 1H), 3.53-3.52 (m, 1H), 1.99 (s, 6H), 1.98 (s, 6H);
.sup.13C-NMR (125 MHz, D.sub.2O) .delta. 119.1, 115.3, 101.9,
100.0, 98.8, 98.5, 76.7, 76.5, 76.3, 76.2, 76.1, 76.0, 7456, 73.3,
73.2, 69.2, 69.0, 68.9, 68.8, 65.9, 55.9, 53.5, 22.0.
[0522] Synthesis of 87
[0523] Compound 87 is prepared from compound 79 following general
procedure L: 15 mg, 69%, R.sub.f=0.1 (EtOAc/ethanol/water 2:2:1).
.sup.13C-NMR (125 MHz, D.sub.2O, HSQC) .delta. 118.4, 115.1, 102.1,
101.1, 96.8, 77.5, 76.6, 76.3, 76.2, 76.1, 76.0, 75.9, 73.6, 73.4,
73.2, 70.6, 69.1, 69.0, 68.8, 68.7, 66.4, 65.8, 55.9, 53.1,
22.0
[0524] Synthesis of 88
[0525] Compound 88 is prepared from compound 80 following general
procedure L: 6 mg, 93% yield, Rf=0.15 (EtOAc:EtOH:water, 2:1:1).
HRMS (ESI) calcd for C.sub.77H.sub.113N.sub.5O.sub.72S.sub.5Na
(M+Na).sup.+ m/z 2442.38, found 2442.0. .sup.1H-NMR (500 MHz,
D.sub.2O) 7.16 (d, J=8.9 Hz, 2H), 7.01 (d, J=8.9 Hz, 2H), 5.47 (d,
J=4.0 Hz, 1H), 5.18 (d, J=4.2 Hz, 1H), 4.99 (d, J=3.5 Hz, 1H), 4.84
(d, J=3.8 Hz, 1H), 4.73-4.69 (m, 3H), 4.36-4.30 (m, 5H), 4.24 (d,
J=11.5 Hz, 1H), 4.18-4.16 (m, 5H), 4.12-4.09 (m, 6H), 4.09-4.03 (m,
10H), 4.03-3.95 (m, 12H), 3.94-3.92 (m, 8H), 3.92-3.86 (m, 6H),
3.81 (s, 3H), 3.78-3.68 (m, 10H), 3.59-3.57 (m, 2H), 3.57-3.55 (m,
2H), 3.54-3.53 (m, 2H), 2.0 (s, 15H); .sup.13C-NMR (125 MHz,
D.sub.2O) .delta. 174.5, 174.4, 154.8, 119.3, 115.2, 101.9, 101.8,
100.7, 100.6, 94.7, 76.8, 74.3, 73.9, 71.1, 70.3, 69.9, 69.8, 69.4,
69.2, 69.1, 68.7, 68.6, 66.2, 55.9, 53.6, 22.0.
[0526] Synthesis of 147
[0527] Compound 147 is prepared from compound 144 following general
procedure L: 26 mg, 9.03 .mu.mol, 91% yield; R.sub.f=0.15
(EtOAc:EtOH:water, 2:1:1); HRMS (ESI) calcd for
C.sub.91H.sub.125N.sub.6O.sub.86S.sub.6Na.sub.9 (M-3H).sup.3- m/z
1024.7598, found: 1024.7605. .sup.1H-NMR (500 MHz, D.sub.2O) 7.16
(d, J=8.1 Hz, 2H), 7.0 (d, J=8.1 Hz, 2H), 5.50 (d, J=3.4 Hz, 1H),
5.19 (bd, 1H), 5.16-5.12 (m, 3H), 5.04-4.99 (m, 3H), 4.9-4.86 (m,
1H), 4.76-4.74 (m, 2H), 4.74-4.68 (m, 5H), 4.68-4.65 (m, 2H),
4.35-4.28 (m, 5H), 4.26-4.18 (m, 6H), 4.16-4.13 (m, 1H), 4.11-4.06
(m, 8H), 4.03-3.97 (m, 10H), 3.97-3.90 (m, 12H), 3.81 (s, 3H),
3.78-3.70 (m, 12H), 3.68-3.64 (m, 2H), 3.64-3.63 (m, 2H), 3.63-3.62
(m, 2H), 3.59-3.58 (m, 1H), 3.57-3.55 (m, 1H), 3.55-3.54 (m, 1H),
3.53-3.50 (m, 1H), 2.01 (s, 18H); .sup.13C-NMR (125 MHz, D.sub.2O)
.delta. 174.5, 173.5, 119.2, 115.2, 101.9, 101.8, 100.6, 95.2,
95.1, 77.2, 74.2, 73.9, 70.0, 69.9, 69.6, 69.4, 69.2, 69.1, 68.9,
68.1, 66.2, 55.9, 53.6, 22.1.
[0528] Synthesis of 89
[0529] Compound 89 is prepared from compound 81 following general
procedure L: 9 mg, 41%, R.sub.f=0.1 (EtOAc/ethanol/water 2:2:1); MS
(ESI, neg. mode) calcd for
C.sub.77H.sub.113N.sub.5O.sub.72S.sub.5Na (M-9Na+8H).sup.- m/z
2440.37, 2441.37, 2442.37, 2443.37, 2444.37, found 2440.38,
2441.37, 2442.40, 2443.38, calcd for
C.sub.77H.sub.113N.sub.5O.sub.72S.sub.5Na (M-10Na+9H).sup.- m/z
2418.39, 2419.39, 2420.39, 2421.39, 2422.39, found 2418.39,
2419.42, 2420.38, 2421.43, 2422.38. .sup.13C-NMR (125 MHz,
D.sub.2O) .delta. 175.4, 175.1, 174.6, 174.4, 154.9, 118.5, 115.2,
102.1, 102.0, 96.8, 77.7, 77.5, 76.8, 76.6, 76.5, 76.4, 76.3, 76.2,
73.6, 70.7, 70.1, 69.1, 68.8, 65.9, 61.3, 55.9, 53.6, 53.1,
22.0
[0530] Synthesis of 90
[0531] Compound 90 is prepared from compound 82 following general
procedure L: 11 mg, 78% yield, Rf=0.15 (EtOAc:EtOH:water, 2:1:1).
HRMS (ESI) calcd for
C.sub.63H.sub.86N.sub.4O.sub.70S.sub.8Na.sub.10 (M+4H).sup.4+ m/z
626.0008, found 626.001. .sup.1H-NMR (500 MHz, D.sub.2O) 7.15 (d,
J=9.2 Hz, 2H), 6.99 (d, J=9.2 Hz, 2H), 5.68 (d, J=3.6 Hz, 1H),
5.18-5.15 (m, 2H), 5.14 (d, J=3.6 Hz, 1H), 5.0-4.93 (m, 2H),
4.89-4.86 (m, 2H), 4.84-4.80 (m, 4H), 4.78 (d, J=3.6 Hz, 1H),
4.73-4.63 (m, 3H), 4.47 (d, J=4.1 Hz, 1H), 4.35-4.26 (m, 6H),
4.26-4.21 (m, 8H), 4.19-4.17 (m, 4H), 4.16-4.15 (m, 2H), 4.12-4.10
(m, 2H), 4.07-4.0 (m, 10H), 4.0-3.93 (m, 8H), 3.92-3.86 (m, 5H),
3.79 (s, 3H), 3.77-3.73 (m, 4H), 3.73-3.68 (m, 2H), 3.68-3.66 (m,
2H), 3.66-3.65 (m, 2H), 3.64 (d, J=2.4 Hz, 1H), 3.62-3.61 (m, 1H),
3.56 (d, J=4.3 Hz, 1H), 3.54 (d, J=4.5 Hz, 1H), 3.52 (d, J=4.6 Hz,
1H), 2.04 (s, 3H), 2.03 (s, 3H), 1.99 (s, 3H), 1.98 (s, 3H);
.sup.13C-NMR (125 MHz, D.sub.2O) .delta. 174.8, 174.5, 119.6,
115.2, 101.9, 101.8, 100.6, 94.2, 74.2, 73.6, 71.1, 70.1, 69.9,
69.4, 69.2, 68.8, 66.5, 55.9, 53.5, 48.9, 22.3, 22.0.
[0532] Synthesis of 148
[0533] Compound 148 is prepared from compound 145 following general
procedure L: 40 mg, 14.18 .mu.mol, 90% yield; R.sub.f=0.15
(EtOAc:EtOH:water, 2:1:1); HRMS (ESI) calcd for C77H113N5O87S10
C.sub.77H.sub.102N.sub.5O.sub.87S.sub.10Na.sub.11 (M-4H).sup.4- m/z
764.7374, found 764.7418. .sup.1H-NMR (500 MHz, D.sub.2O) 7.18 (d,
J=8.3 Hz, 2H), 7.06 (d, J=8.3 Hz, 2H), 5.69 (d, J=3.4 Hz, 1H), 5.18
(bd, 1H), 5.15 (bd, 1H), 4.91 (bd, 1H), 4.76 (d, J=3.8 Hz, 1H),
4.73-4.69 (m, 5H), 4.68 (d, J=3.8 Hz, 1H), 4.36-4.25 (m, 8H), 4.25
(d, J=11.4 Hz, 1H), 4.12-4.09 (m, 3H), 4.07-3.99 (m, 16H),
3.99-3.94 (m, 6H), 3.83-3.80 (m, 3H), 3.81 (s, 3H), 3.80-3.71 (m,
12H), 3.71-3.66 (m, 6H), 3.58-3.52 (m, 4H), 3.57-3.55 (m, 3H),
3.54-3.53 (m, 3H), 2.05 (s, 15H); .sup.13C-NMR (125 MHz, D.sub.2O)
.delta. 175.4, 174.8, 154.8, 119.7, 115.2, 101.9, 99.3, 99.2, 99.1,
93.7, 93.6, 73.8, 71.1, 70.1, 69.4, 67.9, 66.5, 69.2, 69.1, 68.7,
68.6, 55.9, 53.5, 22.3.
[0534] Synthesis of 149
[0535] Compound 149 is prepared from compound 146 following general
procedure L: 25 mg, 7.44 .mu.mol, 84% yield; R.sub.f=0.15
(EtOAc:EtOH:water, 2:1:1); HRMS (ESI) calcd for
C.sub.91H.sub.121N.sub.6O.sub.104S.sub.12Na.sub.13 (M-5H).sup.5-
m/z 728.1862, found 728.1872. .sup.1H-NMR (500 MHz, D.sub.2O) 7.18
(d, J=9.0 Hz, 2H), 7.07 (d, J=9.0 Hz, 2H), 5.71 (d, J=3.4 Hz, 1H),
5.22-5.17 (m, 2H), 5.17-5.12 (m, 3H), 5.07-5.03 (m, 3H), 5.01 (bd,
1H), 4.88 (bd, 1H), 4.76-4.75 (m, 2H), 4.75-4.69 (m, 5H), 4.68-4.65
(m, 2H), 4.50-4.47 (m, 1H), 4.50-4.47 (m, 1H), 4.36-4.32 (m, 3H),
4.32-4.29 (m, 4H), 4.36-4.32 (m, 3H), 4.29-4.25 (m, 5H), 4.25-4.21
(m, 2H), 4.16-4.12 (m, 1H), 4.10-4.06 (m, 8H), 4.06-4.0 (m, 10H),
4.0-3.93 (m, 8H), 3.93-3.88 (m, 4H), 3.82 (s, 3H), 3.80-3.72 (m,
10H), 3.72-3.70 (m, 2H), 3.68-3.66 (m, 2H), 3.66-3.64 (m, 2H),
3.59-3.56 (m, 1H), 3.56-3.55 (m, 1H), 3.54-3.53 (m, 1H), 3.52-3.50
(m, 1H), 2.06 (s, 18H); .sup.13C-NMR (125 MHz, D.sub.2O) .delta.
174.9, 174.0, 119.6, 115.2, 99.9, 99.6, 99.3, 98.8, 98.7, 94.6,
94.5, 94.0, 76.5, 73.9, 71.5, 71.2, 70.3, 69.9, 69.8, 69.6, 69.3,
67.5, 66.5, 64.5, 55.9, 53.4, 22.3.
[0536] Synthesis of 91
[0537] Compound 91 is prepared from compound 83 following general
procedure L: 8 mg, 90%, R.sub.f=0.05 (EtOAc/ethanol/water 2:2:1).
.sup.13C-NMR (125 MHz, D.sub.2O, HSQC) .delta. 118.6, 115.0, 99.9,
99.8, 97.5, 97.1, 80.4, 80.0, 78.0, 77.6, 77.5, 76.8, 76.4, 76.3,
75.4, 74.8, 70.5, 70.0, 69.6, 69.1, 69.0, 68.8, 68.5, 66.3, 65.8,
61.2, 61.1, 55.7, 53.2, 22.0
[0538] Synthesis of 92
[0539] Compound 92 is prepared from compound 81 following general
procedures J followed by L: 6 mg, quant. R.sub.f=0.05
(EtOAc/ethanol/water 2:2:1). .sup.13C-NMR (125 MHz, D.sub.2O)
.delta. 174.8, 174.6, 174.4, 174.3, 155.8, 142.3, 118.7, 115.1,
100.0, 99.9, 97.5, 97.2, 97.1, 96.9, 96.8, 80.0, 77.8, 77.5, 76.5,
76.1, 75.9, 75.5, 75.3, 70.7, 69.1, 68.8, 68.7, 68.6, 65.8, 55.9,
53.3, 22.0
##STR00041## ##STR00042##
##STR00043## ##STR00044##
##STR00045##
[0540] Synthesis of 93
[0541] Compound 93 is prepared from compounds 30 and 22 following
general procedure E.
[0542] .sup.1H NMR; (CDCl.sub.3) .delta. 8.15-8.03 (m, 6H),
7.75-7.05 (m, 47H), 7.03 (d, J=9.2 Hz, 2H), 6.83 (d, J=9.2 Hz, 2H),
5.61 (s, 1H), 5.46 (d, J=4.2 Hz, 1H), 5.37-5.34 (m, 2H), 5.11, (t,
J=4.2 Hz, 1H), 5.02 (d, J=4.0 Hz, 1H), 4.99 (d, J=11.7 Hz, 1H),
4.92-3.50 (m, 56H), 3.34 (dd, J=10.1, 3.9 Hz, 1H), 3.27 (dd,
J=10.1, 3.9 Hz, 1H), 3.22 (dd, J=9.9, 3.9 Hz, 1H), 2.01, 1.99, 1.97
(s, 3H each). .sup.13C NMR; .delta. 170.6, 170.4, 166.9, 166.7,
166.6, 165.7, 165.4, 164.9, 155.2, 154.1, 150.2, 143.2, 143.0,
141.3, 138.2, 137.7, 137.6, 137.3,137.1, 137.1, 133.7, 133.5,
133.4, 129.9, 129.7, 129.4, 129.0, 128.8, 128.7, 128.7, 128.4,
128.3, 128.2, 128.2, 128.1, 128.0, 127.9, 127.6, 127.3, 127.2,
125.3, 125.1, 124.9, 120.1, 118.3, 114.6, 100.9, 98.6, 98.2, 97.8,
97.7, 97.5, 82.7, 79.5, 78.9, 78.1, 77.7, 77.5, 75.5, 75.3, 75.2,
74.9, 74.7, 74.6, 74.1, 73.6, 72.4, 72.3, 72.1, 70.4, 70.3, 69.9,
68.8, 68.3, 67.6, 65.7, 65.0, 64.9, 63.8, 63.2, 63.2, 62.6, 62.2,
61.9, 61.8, 55.7, 46.7, 40.6, 40.5, 40.4, 20.7.
[0543] Synthesis of 94
[0544] Compound 94 is prepared from compound 93 following general
procedure D.
[0545] .sup.1H NMR; (CDCl.sub.3) .delta. 8.15-8.03 (m, 6H),
7.59-7.05 (m, 39H), 7.03 (d, J=9.3 Hz, 2H), 6.84 (d, J=9.3 Hz, 2H),
5.61 (s, 1H), 5.44 (d, J=3.9 Hz, 1H), 5.36-5.32 (m, 2H), 5.10 (t,
J=4.5 Hz, 1H), 5.02 (d, J=4.1 Hz, 1H), 4.98 (d, J=11.7 Hz, 1H),
4.93-4.36 (m, 16H), 4.31-3.38 (m, 37H), 3.29-3.20 (m, 3H), 2.97 (s,
1H), 2.06, 1.98, 1.96 (s, 3H each). .sup.13C NMR; 171.9, 170.6,
170.5, 166.9, 166.7, 166.6, 165.7, 165.4, 165.0, 155.2, 150.2,
138.3, 137.8, 137.7, 137.6, 137.3, 137.2, 133.7, 133.5, 133.4,
129.9, 129.7, 129.4, 129.0, 128.8, 128.7, 128.7, 128.4, 128.4,
128.4, 128.3, 128.2, 128.2, 128.1, 128.1, 128.0, 127.9, 127.8,
127.6, 127.4, 127.2, 118.3, 114.6, 100.9, 98.6, 98.4, 98.2, 97.7,
97.5, 82.8, 79.2, 78.9, 78.0, 77.8, 75.5, 75.4, 75.0, 74.9, 74.7,
74.6, 74.1, 73.5, 72.4, 72.4, 72.1, 71.4, 70.6, 70.4, 70.3, 69.9,
68.3, 67.6, 65.7, 64.9, 64.9, 63.8, 63.3, 63.2, 62.7, 62.6, 62.2,
61.9, 55.7, 40.6, 40.5, 40.4, 20.8, 20.7.
[0546] Synthesis of 95
[0547] Compound 95 is prepared from compounds 94 and 23 following
general procedure E.
[0548] .sup.1H NMR; (CDCl.sub.3) .delta. 8.15-8.04 (m, 8H),
7.58-7.05 (m, 57H), 7.03 (d, J=9.3 Hz, 2H), 6.83 (d, J=9.3 Hz, 2H),
5.61 (s, 1H), 5.45 (d, J=4.2 Hz, 1H), 5.37-5.34 (m, 3H), 5.27 (t,
J=8.2 Hz, 1H), 5.11-5.05 (m, 2H), 5.00-4.95 (m, 2H), 4.91-4.53 (m,
18H), 4.48 (d, J=10.1 Hz, 1H), 4.42 (dd, J=11.5, 8.1 Hz, 1H),
4.34-3.48 (m, 51H), 3.32-3.18 (m, 4H), 2.00, 1.99, 1.98, 1.95 (s,
3H each). .sup.13C NMR; .delta. 170.6, 170.5, 170.4, 166.9, 166.7,
166.6, 166.4, 165.7, 165.4, 165.0, 164.9, 155.2, 150.2, 138.2,
137.7, 137.6, 137.5, 137.3, 137.3, 133.7, 133.5, 133.4, 130.0,
129.9, 129.7, 129.4, 129.0, 128.9, 128.8, 128.7, 128.6, 128.6,
128.5, 128.4, 128.4, 128.3, 128.3, 128.2, 128.1, 128.1, 127.9,
127.8, 127.6, 127.5, 127.4, 127.4, 127.3, 118.3, 114.6, 100.9,
100.8, 98.5, 98.3, 98.1, 97.8, 97.7, 97.5, 82.8, 80.1, 78.9, 77.9,
77.9, 77.8, 77.7, 77.6, 75.6, 75.5, 75.2, 75.1, 74.9, 74.8, 74.7,
74.6, 74.2, 73.6, 72.4, 72.3, 72.1, 70.4, 69.9, 69.8, 68.3, 67.8,
65.7, 65.0, 64.9, 64.5, 63.8, 63.2, 63.2, 63.1, 62.7, 62.4, 62.2,
61.8, 55.7, 40.7, 40.6, 40.5, 40.3, 20.8, 20.7.
[0549] Synthesis of 96
[0550] Compound 96 is prepared from compound 95 following general
procedure F.
[0551] .sup.1H NMR; (CDCl.sub.3) .delta. 8.14-8.04 (m, 8H),
7.58-7.09 (m, 57H), 7.06 (d, J=9.2 Hz, 2H), 6.83 (d, J=9.2 Hz, 2H),
5.63 (s, 1H), 5.52 (d, J=4.2 Hz, 1H), 5.42 (d, J=4.1 Hz, 1H), 5.35
(s, 1H), 5.32 (t, J=8.6 Hz, 1H), 5.26 (t, J=8.5 Hz, 1H), 5.09 (t,
J=2.9 Hz, 1H), 5.04-4.98 (m, 3H), 4.86-3.46 (m, 60H),
3.32-3.24(m,9H), 3.07 (br s, 1H), 2.95 (br s, 1H), 2.00, 1.99,
1.98, 1.97 (s, 3H each). .sup.13C NMR; .delta. 170.6, 170.45,
170.40, 165.78, 165.66, 165.05, 164.94, 155.25, 150.38, 138.14,
137.72, 137.60, 137.49, 137.37, 133.60, 133.48, 133.28, 129.91,
129.83, 129.55, 129.11, 129.08, 128.74, 128.67, 128.57, 128.45,
128.41,128.35, 128.24, 128.15, 128.07, 128.04, 127.95, 127.82,
127.74, 127.62, 127.10, 126.95, 118.34, 114.70, 101.06, 98.09,
98.03, 97.90, 97.63, 97.32, 83.54, 80.28, 79.30, 78.32, 77.99,
77.79, 77.73, 77.64, 75.56, 75.21, 75.14, 74.97, 74.85, 74.54,
74.44, 74.35, 74.17, 73.58, 73.46, 73.39, 72.90, 72.29, 72.11,
70.42, 69.99, 69.60, 69.52, 69.15, 68.89, 67.93, 67.85, 64.08,
63.35, 63.25, 62.86, 62.68, 62.01, 61.81, 61.61, 61.41, 60.92,
55.66, 20.81, 20.76, 20.70.
[0552] Synthesis of 97
[0553] Compound 97 is prepared from compound 96 following general
procedure G2.
[0554] .sup.1H NMR; (CDCl.sub.3) .delta. 8.12-8.02, (m, 8H),
7.61-7.04 (m, 57H), 7.01 (d, J=9.1 Hz, 2H), 6.80 (d, J=9.1 Hz, 2H),
5.69 (s, 1H), 5.46 (d, J=6.3 Hz, 1H), 5.43-5.38 (m, 5H), 5.34 (dd,
J=8.7, 8.1 Hz, 1H), 5.30 (t, J=2.1 Hz, 1H), 5.17-5.14 (m, 2H),
4.98-4.54 (m, 19H), 4.43 (d, J=10.5 Hz, 1H), 4.37 (d, J=10.9 Hz,
1H), 4.31-4.08 (m, 11H), 4.05 (t, J=2.8 Hz, 1H), 4.01 (t, J=7.2 Hz,
1H), 3.94-3.65 (m, 16H), 3.59-3.42 (m, 12H), 3.34-3.30 (m, 2H),
3.24-3.17 (m, 3H), 2.97 (s, 3H), 2.09, 2.07, 2.01, 2.00 (s, 3H
each). .sup.13C NMR; .delta. 170.75, 170.61, 170.38, 169.67,
169.16, 167.70, 167.50, 165.62, 165.10, 164.66, 155.25, 150.49,
138.12, 137.86, 137.57, 137.52, 137.40, 137.17, 137.07, 133.92,
133.73, 133.64, 133.53, 129.97, 129.90, 129.71, 129.47, 129.13,
129.05, 128.94, 128.86, 128.80, 128.76, 128.55, 128.41, 128.33,
128.24, 128.16, 128.06, 128.03, 127.93, 127.77, 127.63, 127.51,
127.46, 125.32, 117.97, 114.66, 101.15, 101.08, 98.89, 98.58,
98.17, 97.83, 97.67, 97.22, 82.81, 82.56, 80.22, 78.31, 77.80,
77.64, 77.51, 77.48, 76.05, 75.58, 75.45, 75.37, 75.28, 75.01,
74.96, 74.86, 74.81, 74.60, 74.35, 73.68, 73.51, 72.86, 72.47,
72.09, 71.45, 69.85, 69.78, 69.38, 69.08, 68.10, 63.41, 63.32,
62.82, 62.73, 62.20, 61.57, 61.42, 55.66, 52.72, 52.08, 51.96,
51.75, 20.87, 20.81.
[0555] Synthesis of 98
[0556] Compound 98 is prepared from compound 97 following general
procedure H.
[0557] .sup.1H NMR; (CDCl.sub.3) .delta. 8.07-7.99 (m, 8H),
7.60-7.41 (m, 12H), 7.35-6.98 (m, 47H), 6.79 (d, J=9.1 Hz, 2H),
5.79 (d, J=9.6 Hz, 1H), 5.72 (d, J=9.6 Hz, 1H), 5.65 (d, J=1.6 Hz,
1H), 5.54 (d, J=9.5 Hz, 1H), 5.42 (t, J=8.4 Hz, 1H), 5.39-5.29 (m,
3H), 5.15 (t, J=5.4 Hz, 1H), 4.99-4.96 (m, 2H), 4.94-4.91 (m, 3H),
4.88-3.90 (m, 44H), 3.86-3.59 (m, 9H), 3.74 (s, 3H), 3.57-3.47 (m,
2H), 3.55, 3.54 (s, 3H each), 3.38-3.32 (m, 1H), 3.36, 3.24, 2.10,
2.01, 1.97, 1.91, 1.40, 1.39, 1.38, 1.31 (s, 3H each). .sup.13C
NMR; .delta. 171.11, 170.70, 170.64, 170.58, 170.32, 170.12,
170.02, 169.28, 169.01, 167.41, 167.20, 165.59, 165.31, 164.87,
164.67, 155.39, 150.41, 138.72, 138.45, 138.12, 137.85, 137.70,
137.03, 136.81, 136.38, 136.32, 133.97, 133.87, 133.81, 129.73,
129.04, 128.98, 128.85, 128.72, 128.62, 128.55, 128.49, 128.32,
128.24, 128.12, 128.09, 128.02, 127.95, 127.78, 127.72, 127.60,
127.50, 127.41, 127.36, 127.30, 127.23, 127.02, 125.31, 118.07,
117.81, 114.68, 101.32, 101.12, 99.50, 98.72, 98.08, 97.81, 97.57,
81.57, 81.40, 80.50, 78.10, 77.89, 77.82, 76.75, 75.42, 75.27,
75.18, 75.02, 74.95, 74.87, 74.73, 74.34, 73.76, 73.55, 72.91,
72.70, 72.56, 71.86, 70.86, 70.70, 70.20, 70.10, 69.77, 68.77,
68.33, 62.19, 61.67, 61.53, 61.38, 55.64, 52.82, 52.52, 52.48,
52.09, 51.97, 51.91, 51.78, 22.76, 22.64, 21.45, 20.98, 20.79,
20.76, 20.71.
[0558] Synthesis of 99
[0559] Compound 99 is prepared from compound 98 following general
procedure I.
[0560] .sup.1H NMR; (CDCl.sub.3) .delta. 8.03-7.92 (m, 8H),
7.58-7.40 (m, 12H), 7.35-6.95 (m, 47H), 6.78 (d, J=9.1 Hz, 2H),
5.75 (d, J=9.0 Hz, 1H), 5.69 (d, J=9.5 Hz, 1H), 5.63 (d, J=2.0 Hz,
1H), 5.52 (d, J=9.1 Hz, 1H), 5.43 (t, J=8.3 Hz, 1H), 5.37-5.32 (m,
3H), 5.28 (d, J=8.5 Hz, 1H), 5.19 (t, J=4.5 Hz, 1H), 5.07 (d, J=3.5
Hz, 1H), 5.00-4.77 (m, 10 H), 4.71-4.45 (m, 11H), 4.32 (d, J=12.2
Hz, 1H), 4.22 (dt, J=10.0, 3.6 Hz, 1H), 4.17-3.19 (m, 36H), 3.74,
3.56, 3.54, 3.24, 3.23, 1.47, 1.45, 1.43, 1.33 (s, 3H each).
.sup.13C NMR; .delta. 170.58, 170.19, 170.09, 170.04, 169.09,
169.05, 167.93, 165.55, 165.38, 165.05, 164.74, 155.39, 150.46,
138.97, 138.92, 138.50, 138.29, 137.93, 137.06, 136.95, 136.54,
133.98, 133.86, 129.75, 129.60, 129.06, 128.97, 128.88, 128.61,
128.57, 128.52, 128.48, 128.31, 128.23, 128.11, 127.98, 127.88,
127.82,127.7, 127.41, 127.24, 118.07, 114.68, 100.64, 100.56,
98.96, 98.36, 98.27, 98.15, 97.74, 97.04, 81.41, 81.28, 80.06,
77.77, 77.54, 77.27, 76.34, 75.50, 75.16, 75.09, 74.98, 74.78,
74.68, 74.44, 74.07, 73.93, 73.72, 73.34, 73.06, 72.91, 72.79,
72.63, 72.49, 72.39, 71.94, 71.44, 70.55, 68.79, 68.48, 61.60,
60.46, 60.40, 60.10, 59.96, 55.66, 52.81, 52.64, 52.32, 52.07,
51.77, 22.66.
[0561] Synthesis of 100
[0562] Compound 100 is prepared from compound 99 following general
procedures J and then K.
[0563] .sup.1H NMR; (MeOD) .delta. 7.44-7.17 (m, 45H), 7.09 (d,
J=9.1 Hz, 2H), 6.84 (d, J=9.1 Hz, 2H), 5.49 (s, 1H), 5.43 (m, 2H),
5.27 (s, 1H), 5.12-4.53 (m, 32H), 4.44-3.51 (m, 34H), 3.73 (s, 3H),
1.86, 1.84, 1.78, 1.66 (s, 3H each). .sup.13C NMR; .delta. 175.40,
175.31, 174.45, 173.33, 173.25, 173.18, 173.11, 168.34, 156.54,
152.41, 140.30, 140.21, 140.04, 139.85, 139.80, 139.45, 139.21,
133.67, 130.69, 129.75, 129.60, 129.49, 129.41, 129.35, 129.29,
129.01, 128.89, 128.69, 128.60, 128.51, 128.47, 128.21, 119.28,
115.72, 103.97, 103.91, 101.99, 101.73, 98.75, 98.22, 97.35, 86.11,
85.74, 81.96, 80.94, 80.50, 80.22, 79.28, 77.93, 77.82, 77.68,
77.56, 76.67, 76.47, 76.30, 76.20, 75.97, 75.83, 75.28, 75.16,
75.09, 74.06, 73.63, 73.15, 72.90, 71.64, 71.40, 71.09, 70.11,
69.31, 68.42, 67.47, 67.30, 67.06, 56.19, 54.38, 53.56, 53.47,
23.00.
[0564] Synthesis of 101
[0565] Compound 101 is prepared from compound 100 following general
procedure L.
[0566] .sup.1H NMR; (D.sub.2O) .delta. 7.13 (d, J=8.8 Hz, 2H), 6.98
(d, J=8.8 Hz, 2H), 5.46 (s, 1H), 5.41 (m, 2H), 5.16 (m, 2H), 4.98
(s, 1H), 4.79-4.68 (m, 4H), 4.57 (m, 2H), 4.44 (m, 2H), 4.36-4.33
(m, 2H), 4.23-4.15 (m, 4H), 4.11 (s, 1H), 4.07-3.69 (m, 24H), 3.80
(s, 3H), 3.56 (t, J=9.6 Hz, 1H), 3.36-3.32 (m, 2H), 2.04, 2.03 (s,
3H each), 2.01 (s, 6H). .sup.13C NMR .delta. 174.97, 174.89,
174.67, 174.52, 174.43, 154.81, 150.34, 119.34, 115.18, 102.00,
101.96, 100.69, 97.08, 96.84, 94.76, 94.39, 77.37, 77.26, 76.90,
76.51, 76.28, 76.20, 74.34, 73.92, 73.57, 70.66, 70.12, 69.95,
69.59, 69.38, 69.21, 69.17, 69.08, 69.01, 68.85, 68.11, 66.41,
66.26, 65.85, 55.91, 53.67, 53.60, 53.14, 53.04, 22.04, 22.01.
[0567] Synthesis of 102
[0568] Compound 102 is prepared from compounds 125 and 24 following
general procedure E.
[0569] .sup.1H NMR; (CDCl.sub.3) .delta. 8.11-8.05 (m, 6H), 7.76
(m, 2H), 7.60-7.17 (m, 43H), 7.08-7.06 (m, 2H), 6.88 (d, J=9.1 Hz,
2H), 6.76 (d, J=9.1 Hz, 2H), 5.54-5.51 (m, 2H), 5.18-5.12 (m, 4H),
5.07 (d, J=7.1 Hz, 1H), 4.96 (d, J=10.4 Hz, 1H), 4.86-4.67 (m,
10H), 4.59 (dd, J=11.8, 2.6 Hz, 1H), 4.51-4.45 (m, 3H), 4.39-3.61
(m, 55H), 3.74 (s, 3H), 3.33-3.27 (m, 3H), 2.04, 2.03, 2.00 (s, 3H
each). .sup.13C NMR; .delta. 170.57, 170.52, 170.46, 166.82,
166.76, 165.51, 165.45, 165.08, 155.70, 154.17, 150.99, 143.20,
143.05, 141.34, 137.85, 137.72, 137.35, 137.24, 137.13, 137.06,
133.54, 133.47, 129.82, 129.52, 129.38, 128.75, 128.67, 128.59,
128.48, 128.44, 128.40, 128.30, 128.26, 128.19, 128.10, 128.02,
127.99, 127.87, 127.79, 127.67, 127.26, 125.03, 124.84, 120.14,
118.77, 114.56, 100.11, 98.81, 98.14, 97.98, 97.94, 97.79, 82.75,
78.84, 78.52, 77.92, 75.59, 75.28, 75.08, 75.01, 74.58, 74.53,
74.29, 74.21, 74.13, 73.87, 73.63, 73.54, 73.38, 72.54, 70.45,
70.32, 70.22, 69.80, 68.83, 67.73, 67.12, 64.60, 63.77, 63.68,
63.61, 63.28, 63.12, 62.19, 61.93, 55.65, 46.75, 40.64, 40.53,
20.77, 20.73, 20.63.
[0570] Synthesis of 103
[0571] Compound 103 is prepared from compound 102 following general
procedure D.
[0572] .sup.1H NMR; (CDCl.sub.3) .delta. 8.12-8.05 (m, 6H), 7.57
(t, J=7.4 Hz, 1H), 7.53-7.48 (m, 2H), 7.46-7.40 (m, 6H), 7.34-7.19
(m, 30H), 6.88 (d, J=9.1 Hz, 2H), 6.76 (d, J=9.1 Hz, 2H), 5.54-5.51
(m, 2H), 5.18-5.14 (m, 2H), 5.12-5.10 (m, 2H), 5.07 (d, J=7.1 Hz,
1H), 4.95 (d, J=10.5 Hz, 1H), 4.86-4.67 (m, 10H), 4.61-4.55 (m,
2H), 4.51-4.44 (m, 3H), 4.41 (d, J=7.8 Hz, 1H), 4.37-4.19 (m, 8H),
4.15-3.98 (m, 8H), 3.94 (s, 2H), 3.91 (s, 2H), 3.89 (s, 2H),
3.91-3.69 (m, 8H), 3.74 (s, 3H), 3.63-3.55 (m, 2H), 3.40-3.27 (m,
3H), 3.22 (dd, J=10.1, 3.6 Hz, 1H), 2.04 (s, 6H), 2.03, 2.02 (s, 3H
each). .sup.13C NMR; .delta. 171.70, 170.70, 170.58, 167.02,
166.79, 166.75, 165.51, 165.08, 155.68, 150.98, 137.84, 137.74,
137.69, 137.34, 137.24, 137.20, 133.52, 133.47, 129.87, 129.81,
129.50, 129.37, 128.69, 128.58, 128.42, 128.40, 128.31, 128.24,
128.16, 128.10, 127.97, 127.78, 127.66, 118.76, 114.55, 100.11,
98.95, 98.23, 97.91, 97.79, 82.74, 79.72, 78.71, 78.50, 75.57,
75.38, 75.27, 75.19, 75.12, 74.65, 74.57, 74.28, 74.23, 73.62,
73.52, 73.43, 72.53, 71.29, 70.56, 70.48, 70.38, 70.31, 70.23,
67.76, 67.68, 64.59, 63.59, 63.27, 63.08, 62.80, 62.17, 60.39,
55.64, 40.66, 40.53, 20.74.
[0573] Synthesis of 104
[0574] Compound 104 is prepared from compounds 103 and 23 following
general procedure E.
[0575] .sup.1H NMR; (CDCl.sub.3) .delta. 8.08-8.00 (m, 8H),
7.58-7.12 (m, 57H), 6.88 (d, J=9.1 Hz, 2H), 6.76 (d, J=9.1 Hz, 2H),
5.54-5.51 (m, 2H), 5.43 (d, J=3.8 Hz, 1H), 5.35 (t, J=8.2 Hz, 1H),
5.16 (t, J=4.3 Hz, 1H), 5.12-5.09 (m, 2H), 5.07 (d, J=7.1 Hz, 1H),
5.03 (d, J=3.8 Hz, 1H), 4.94 (d, J=10.4 Hz, 1H), 4.91-4.54 (m,
14H), 4.44 (dd, J=10.4, 7.6 Hz, 1H), 4.34-3.55 (m, 41H), 3.93,
3.89, 3.81, 3.80 (s, 2H each), 3.73 (s, 3H), 3.49 (t, J=9.4 Hz,
1H), 3.32-3.20 (m, 4H), 2.03, 2.00, 1.99, 1.96 (s, 3H each).
.sup.13C NMR; .delta. 170.58, 170.52, 170.43, 166.82, 166.79,
166.75, 166.60, 165.48, 165.43, 165.09, 164.94, 155.70, 150.99,
138.19, 137.84, 137.72, 137.51, 137.35, 137.25, 137.18, 133.70,
133.47, 129.83, 129.51, 129.36, 129.06, 128.80, 128.75, 128.68,
128.62, 128.57, 128.40, 128.29, 128.25, 128.19, 128.12, 128.04,
127.98, 127.79, 127.68, 127.56, 127.21, 125.33, 118.77, 114.56,
100.95, 100.11, 98.57, 98.20, 98.00, 97.91, 97.81, 82.74, 80.13,
78.66, 78.49, 78.06, 77.90, 77.75, 75.64, 75.57, 75.45, 75.23,
75.09, 74.87, 74.77, 74.60, 74.43, 74.22, 74.11, 73.62, 73.56,
72.54, 72.32, 70.49, 70.31, 69.89, 67.81, 67.62, 65.02, 64.58,
63.57, 63.50, 63.21, 62.39, 62.16, 61.88, 55.65, 40.65, 40.60,
40.53, 40.45, 21.46, 20.78, 20.70.
[0576] Synthesis of 105
[0577] Compound 105 is prepared from compound 104 following general
procedure F.
[0578] .sup.1H NMR; (CDCl.sub.3) .delta. 8.10-8.00 (m, 8H),
7.58-7.49 (m, 3H), 7.46-7.12 (m, 54H), 6.89 (d, J=9.0 Hz, 2H), 6.76
(d, J=9.0 Hz, 2H), 5.65 (s, 1H), 5.54 (t, J=8.0 Hz, 1H), 5.50 (s,
1H), 5.31 (t, J=8.1 Hz, 1H), 5.17 (s, 1H), 5.12-5.08 (m, 3H), 4.99
(s, 1H), 4.89-4.63 (m, 12H), 4.58-4.53 (m, 3H), 4.47-4.43 (m, 2H),
4.35-3.42 (m, 40H), 3.72 (s, 3H), 3.35-3.24 (m, 6H), 3.16-3.11 (m,
1H), 3.06-3.01 (m, 1H), 2.03 (s, 3H), 1.99 (s, 6H), 1.97 (s, 3H).
.sup.13C NMR; .delta. 170.58, 170.42, 165.82, 165.72, 165.10,
164.96, 155.65, 151.13, 138.14, 137.88, 137.61, 137.53, 137.46,
137.34, 133.61, 133.40, 129.81, 129.63, 129.06, 128.75, 128.68,
128.55, 128.45, 128.37, 128.26, 128.14, 128.08, 128.03, 127.86,
127.77, 127.65, 126.86, 125.33, 118.51, 114.66, 101.22, 100.53,
98.12, 97.94, 97.86, 97.62, 97.42, 83.65, 83.35, 80.27, 79.21,
78.82, 78.33, 78.00, 77.78, 77.34, 77.08, 76.83, 75.55, 75.14,
75.07, 74.88, 74.81, 74.46, 74.36, 74.30, 74.17, 73.97, 73.42,
73.35, 73.06, 72.99, 72.79, 72.49, 70.22, 70.09, 69.96, 69.62,
68.82, 68.19, 67.53, 63.98, 63.46, 63.24, 62.63, 62.43, 62.10,
61.95, 61.77, 61.22, 61.02, 55.65, 21.47, 20.81.
[0579] Synthesis of 106
[0580] Compound 106 is prepared from compound 105 following general
procedure G2.
[0581] .sup.1H NMR; (CDCl.sub.3) .delta. 8.12-8.01 (m, 8H),
7.58-7.52 (m, 4H), 7.48-7.41 (m, 8H), 7.37-7.10 (m, 45H), 6.85 (d,
J=9.1 Hz, 2H), 6.73 (d, J=9.1 Hz, 2H), 5.52-5.39 (m, 6H), 5.18-5.14
(m, 2H), 5.07 (d, J=6.8 Hz, 1H), 5.03 (d, J=10.8 Hz, 1H), 4.94 (d,
J=3.6 Hz, 1H), 4.91-4.86 (m, 4H), 4.82 (s, 1H), 4.80 (s, 1H),
4.78-4.63 (m, 9H), 4.55 (d, J=11.0 Hz, 1H), 4.44 (d, J=10.8 Hz,
1H), 4.37-4.18 (m, 9H), 4.14-4.01 (m, 8H), 3.97-3.69 (m, 11H), 3.72
(s, 3H), 3.59-3.42 (m, 5H), 3.51, 3.50, 3.43 (s, 3H each),
3.34-3.21 (m, 4H), 3.26 (s, 3H), 2.09, 2.08, 2.02, 2.01 (s, 3H
each). .sup.13C NMR; .delta. 170.76, 170.68, 170.60, 170.42,
169.73, 169.57, 168.14, 167.70, 165.22, 165.09, 165.00, 164.69,
155.70, 150.95, 137.96, 137.89, 137.76, 137.57, 137.52, 137.44,
137.39, 137.29, 137.19, 133.92, 133.81, 133.63, 133.44, 129.95,
129.86, 129.80, 129.74, 129.46, 129.11, 129.05, 128.86, 128.77,
128.55, 128.33, 128.25, 128.16, 128.06, 127.94, 127.85, 127.80,
127.59, 125.32, 118.74, 114.50, 101.19, 100.73, 98.72, 98.66,
97.91, 97.85, 97.69, 97.40, 82.49, 82.17, 80.23, 77.99, 77.74,
77.63, 77.48, 76.28, 75.95, 75.58, 75.33, 75.10, 75.02, 74.93,
74.84, 74.65, 74.57, 74.51, 74.37, 74.26, 73.80, 73.67, 72.28,
71.62, 71.15, 69.87, 69.65, 69.45, 63.42, 62.95, 62.84, 62.19,
61.55, 55.62, 52.70, 52.22, 51.85, 51.75, 20.80, 20.75.
[0582] Synthesis of 107
[0583] Compound 107 is prepared from compound 106 following general
procedure H.
[0584] .sup.1H NMR; (CDCl.sub.3) .delta. 8.07-8.00 (m, 8H),
7.59-7.54 (m, 4H), 7.47-7.42 (m, 6H), 7.35-7.07 (m, 47H), 6.84 (d,
J=9.1 Hz, 2H), 6.73 (d, J=9.1 Hz, 2H), 5.72-5.68 (m, 2H), 5.64-5.61
(m, 2H), 5.53 (dd, J=8.5, 6.6 Hz, 1H), 5.42 (dd, J=9.0, 8.0 Hz,
1H), 5.33-5.30 (m, 2H), 5.20-5.15 (m, 2H), 5.06 (d, J=6.6 Hz, 1H),
5.05 (d, J=3.4, Hz, 1H), 4.96-4.88 (m, 5H), 4.84-4.47 (m, 18H),
4.41-4.35 (m, 4H), 4.32-3.91 (m, 19H), 3.84-3.46 (m, 10H), 3.72,
3.55, 3.49 (s, 3H each), 3.41-3.35 (m, 2H), 3.37, 3.32 (s, 3H
each), 2.09, 2.08, 2.01, 1.94, 1.39 (s, 3H each), 1.38 (s, 6H),
1.35 (s, 3H). .sup.13C NMR; .delta. 171.04, 170.67, 170.62. 170.21,
170.06, 169.99, 169.28, 169.16, 167.69, 167.22, 165.49, 165.21,
164.99, 164.72, 155.74, 150.86, 138.70, 138.47, 138.13, 137.70,
136.88, 136.77, 136.51, 136.37, 133.85, 133.58, 129.76, 129.70,
129.26, 129.03, 128.95, 128.62, 128.53, 128.49, 128.31, 128.23,
128.16, 128.10, 128.02, 127.90, 127.78, 127.71, 127.56, 127.37,
127.29, 127.06, 125.30, 118.61, 114.51, 101.36, 100.90, 99.62,
99.27, 98.02, 97.83, 81.40, 80.93, 80.52, 78.02, 77.90, 77.49,
75.75, 75.45, 75.31, 74.96, 74.86, 74.67, 73.93, 73.86, 73.58,
73.21, 72.08, 71.64, 71.06, 70.72, 70.65, 70.15, 69.82, 62.18,
61.52, 55.61, 52.79, 52.71, 52.53, 51.76, 51.69, 22.75, 22.71,
22.61, 22.53, 20.97, 20.95, 20.79, 20.73.
[0585] Synthesis of 108
[0586] Compound 108 is prepared from compound 107 following general
procedure I.
[0587] .sup.1H NMR; (CDCl.sub.3) .delta. 8.04-8.00 (m, 4H),
7.96-7.92 (m, 4H), 7.60-7.54 (m, 4H), 7.48-7.41 (m, 8H), 7.35-7.09
(m, 45H), 6.84 (d, J=9.1 Hz, 2H), 6.73 (d, J=9.1 Hz, 2H), 5.99 (d,
J=9.2 Hz, 1H), 5.91 (d, J=9.1 Hz, 1H), 5.75 (d, J=9.5 Hz, 1H), 5.63
(d, J=9.3 Hz, 1H), 5.53 (dd, J=8.5, 6.8 Hz, 1H), 5.45-5.39 (m, 2H),
5.34 (d, J=4.8 Hz, 1H), 5.21 (t, J=6.0 Hz, 1H), 5.14 (br s, 1H),
5.12 (d, J=3.5 Hz, 1H), 5.07 (d, J=6.7 Hz, 1H), 4.99 (d, J=3.5 Hz,
1H), 4.96-4.90 (m, 3H), 4.83-4.73 (m, 5H), 4.68-4.49 (m, 12H),
4.37-4.34 (m, 2H), 4.28-4.20 (m, 4H), 4.14-3.85 (m, 16H), 3.73-3.16
(m, 15H), 3.71, 3.58, 3.49, 3.35, 3.20, 1.46, 1.44, 1.44, 1.41 (s,
3H each). .sup.13C NMR .delta. 170.72, 170.53, 170.27, 170.12,
169.15, 169.09, 167.97, 167.91, 165.55, 165.41, 165.02, 164.78,
155.73, 150.85, 138.96, 138.50, 138.45, 138.29, 137.94, 137.00,
136.87, 136.54, 133.85, 133.59, 129.74, 129.65,129.55, 129.23,
128.98, 128.85, 128.59, 128.53, 128.34, 128.25, 128.14, 127.97,
127.92, 127.83, 127.78, 127.59, 127.37, 127.22, 127.16, 118.61,
114.52, 100.88, 100.75, 98.92, 97.63, 96.87, 96.66, 81.34, 81.07,
80.09, 77.71, 77.57, 76.75, 76.39, 76.30, 76.26, 75.43, 75.36,
75.19, 75.00, 74.76, 74.66, 74.31, 73.93, 73.77, 73.63, 73.33,
73.10, 72.94, 72.39, 71.98, 71.27, 71.10, 70.23, 67.19, 67.14,
66.53, 66.27, 66.19, 61.57, 60.51, 60.33, 59.91, 55.60, 52.73,
52.64, 52.12, 51.83, 51.76, 22.67, 22.62.
[0588] Synthesis of 109
[0589] Compound 109 is prepared from compound 108 following general
procedure J.
[0590] .sup.1H NMR; (CD.sub.3OD) .delta. 8.22 (d, J=7.3 Hz, 2H),
8.12-8.09 (m, 4H), 8.04 (d, J=7.3 Hz, 2H), 7.65-7.43 (m, 12H),
7.35-7.01 (m, 43H), 6.97-6.95 (m, 2H), 6.85 (d, J=9.1 Hz, 2H), 6.75
(d, J=9.1 Hz, 2H), 5.58 (d, J=3.7 Hz, 1H), 5.53 (d, J=4.6 Hz, 1H),
5.43-5.35 (m, 4H), 5.28-5.26 (m, 2H), 5.23 (t, J=4.7 Hz, 1H), 5.17
(t, J=5.5 Hz, 1H), 5.04 (d, J=3.2 Hz, 1H), 5.01 (d, J=11.0 Hz, 1H),
4.97 (d, J=3.4 Hz, 1H), 4.86-4.14 (m, 35H), 4.09-3.90 (m, 8H),
3.74-3.48 (m, 7H), 3.63, 3.56, 3.50 (s, 3H each), 3.44-3.39 (m,
3H), 3.19, 2.98, 1.85, 1.77, 1.55, 1.47 (s, 3H each). .sup.13C NMR;
(CD.sub.3OD) .delta. 173.53, 173.33, 173.24, 170.78, 170.49,
170.43, 170.38, 167.43, 167.26, 166.86, 166.56, 157.24, 152.36,
140.26, 140.07, 139.87, 139.82, 138.79, 138.67, 138.49, 135.11,
134.84, 131.23, 130.86, 130.61, 130.16, 130.04, 129.95, 129.86,
129.58, 129.51, 129.44, 129.36, 129.31, 129.28, 129.22, 129.11,
129.04, 128.93, 128.84, 128.59, 128.40, 128.25, 128.14, 119.51,
115.73, 101.70, 99.18, 99.08, 98.95, 97.45, 97.01, 83.95, 83.43,
81.72, 79.90, 79.60, 79.36, 78.96, 78.19, 76.87, 76.22, 75.87,
75.65, 75.41, 75.31, 75.14, 74.98, 74.79, 74.59, 74.46, 73.90,
72.82, 72.27, 71.79, 71.42, 71.13, 70.63, 66.83, 66.21, 65.74,
56.14, 54.07, 53.85, 53.66, 53.51, 53.33, 52.82, 52.58, 22.93,
22.84, 22.80.
[0591] Synthesis of 110
[0592] Compound 110 is prepared from compound 109 following general
procedure K.
[0593] .sup.1H NMR; (CD.sub.3OD) .delta. 7.39-7.18 (m, 45H), 7.04
(d, J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.42-5.40 (m, 2H), 5.23
(br s, 2H), 5.11-5.07 (m, 2H), 5.01-4.54 (m, 25H), 4.47-3.51 (m,
35H), 3.74, 1.86, 1.77, 1.69, 1.62 (s, 3H each). .sup.13C NMR;
(CD.sub.3OD) .delta. 175.75, 175.05, 173.29, 173.12, 173.00,
156.89, 153.22, 140.26, 139.98, 139.86, 139.73, 139.33, 139.19,
129.76, 129.60, 129.54, 129.49, 129.40, 129.32, 129.03, 128.86,
128.80, 128.63, 128.58, 119.55, 115.57, 103.94, 103.85, 102.28,
102.09, 98.82, 98.63, 98.03, 97.13, 86.19, 85.83, 81.96, 80.96,
80.84, 80.48, 79.34, 78.33, 78.04, 77.62, 76.62, 76.30, 75.95,
75.85, 75.65, 75.43, 75.16, 74.78, 73.99, 73.15, 71.69, 71.46,
71.35, 70.59, 70.18, 69.39, 67.41, 67.08, 56.16, 54.41, 54.30,
53.60, 23.00.
[0594] Synthesis of 111
[0595] Compound 111 is prepared from compound 110 following general
procedure L.
[0596] .sup.1H NMR; (D.sub.2O) .delta. 7.10 (d, J=8.3 Hz, 2H), 6.97
(d, J=8.3 Hz, 2H), 5.42-5.39 (m, 2H), 5.15 (br s, 2H), 4.98-4.96
(m, 3H), 4.76-4.74 (m, 2H and HOD), 4.57 (d, J=7.8 Hz, 1H), 4.45
(d, J=10.8 Hz, 1H), 4.36-4.31 (m, 3H), 4.22-4.14 (m, 4H), 4.08-3.69
(m, 27H), 3.58-3.55 (m, 2H), 3.36-3.32 (m, 1H), 2.04, 2.00 (s, 6H
each). .sup.13C NMR .delta. 175.05, 174.95, 174.67, 174.52, 174.42,
154.84, 151.02, 118.38, 115.13, 102.01, 101.96, 101.91, 101.29,
97.17, 97.05, 94.48, 94.35, 77.25, 77.11, 76.92, 76.80, 76.70,
76.62, 76.44, 76.30, 76.20, 74.40, 74.22, 73.53, 73.35, 70.66,
70.10, 69.96, 69.60, 69.55, 69.41, 69.35, 69.31, 69.23, 69.13,
68.95, 68.79, 66.38, 66.23, 65.88, 55.90, 53.76, 53.65, 53.59,
53.04, 22.00.
[0597] Synthesis of 112
[0598] Compound 112 is prepared from compounds 26 and 22 following
general procedure E.
[0599] .sup.1H NMR; (CDCl.sub.3) .delta. 8.13-8.08 (m, 4H),
7.76-7.73 (m, 2H), 7.60-7.53 (m, 4H), 7.49-7.33 (m, 10H), 7.30-7.13
(m, 18H), 7.01 (d, J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.57 (s,
1H), 5.48 (d, J=3.9 Hz, 1H), 5.36 (t, J=8.3 Hz, 1H), 5.33 (t, J=1.6
Hz, 1H), 4.97 (d, J=11.7 Hz, 1H), 4.86 (t, J=9.5 Hz, 1H), 4.78-4.69
(m, 4H), 4.65-4.61 (m, 2H), 4.57 (d, J=3.8 Hz, 1H), 4.49-4.45 (m,
2H), 4.33-4.13 (m, 10H), 4.05-3.58 (m, 15H), 3.77 (s, 3H), 3.51 (t,
J=9.4 Hz, 1H), 3.35 (dd, J=10.2, 3.9 Hz, 1H), 3.25 (dd, J=10.1, 3.8
Hz, 1H), 2.02, 1.98 (s, 3H each). .sup.13C NMR .delta. 170.55,
170.45, 166.87, 166.56, 165.65, 164.93, 155.23, 154.12, 150.12,
143.25, 143.01, 141.32, 138.25, 137.59, 137.19, 137.09, 133.67,
133.55, 129.89, 129.85, 129.63, 129.05, 129.02, 128.81, 128.76,
128.42, 128.25, 128.12, 128.09, 127.98, 127.93, 127.87, 127.57,
127.48, 127.25, 127.17, 125.32, 125.11, 124.91, 120.12, 118.29,
114.56, 100.88, 98.76, 97.85, 97.51, 82.67, 78.40, 77.78, 77.49,
75.43, 75.21, 74.69, 74.61, 74.57, 74.11, 72.44, 72.38, 72.28,
70.40, 69.99, 68.77, 68.09, 65.60, 65.01, 64.76, 63.34, 62.63,
62.15, 61.80, 55.70, 46.71, 40.44, 40.41, 20.69.
[0600] Synthesis of 113
[0601] Compound 113 is prepared from compound 112 following general
procedure D.
[0602] .sup.1NMR; (CDCl.sub.3) .delta.8.13-8.08 (m, 4H), 7.57-7.52
(m, 2H), 7.49-7.13 (m, 24H), 7.01 (d, J=9.1 Hz, 2H), 6.83 (d, J=9.1
Hz, 2H), 5.57 (s, 1H), 5.45 (d, J=3.9 Hz, 1H), 5.36 (t, J=8.2 Hz,
1H), 5.32 (s, 1H), 4.97 (d, J=11.6 Hz, 1H), 4.89 (s, 2H), 4.80-4.68
(m, 4H), 4.60 (d, J=7.8 Hz, 1H), 4.57 (d, J=3.7 Hz, 1H), 4.51-4.45
(m, 2H), 4.26-4.10 (m, 5H), 4.06 (d, J=10.4 Hz, 1H), 4.00-3.96 (m,
2H), 3.90 (d, J=11.0 Hz, 1H), 3.85-3.58 (m, 12H), 3.78 (s, 3H),
3.51 (t, J=10 Hz, 1H), 3.44-3.39 (m, 1H), 3.25-3.22 (m, 2H), 2.95
(d, J=4 Hz, 1H), 2.07, 1.97 (s, 3H each). .sup.13C NMR .delta.
171.87, 170.56, 166.90, 166.54, 165.64, 164.94, 155.21, 150.11,
138.29, 137.78, 137.58, 137.25, 133.65, 133.54, 129.88, 129.84,
129.62, 129.03, 128.79, 128.76, 128.66, 128.40, 128.20, 128.16,
128.11, 128.07, 127.97, 127.79, 127.61, 127.44, 127.10, 118.27,
114.54, 100.87, 98.77, 98.25, 97.49, 82.78, 79.16, 78.38, 77.81,
75.42, 75.14, 74.58, 74.16, 72.40, 71.35, 70.61, 69.98, 68.05,
65.58, 64.97, 64.74, 63.32, 62.71, 62.66, 62.15, 55.69, 40.43,
20.78, 20.70.
[0603] Synthesis of 114
[0604] Compound 114 is prepared from compounds 113 and 24 following
general procedure E.
[0605] .sup.1H NMR; (CDCl.sub.3) .delta. 8.13-8.07(m, 6H),
7.78-7.75 (m, 2H), 7.60-7.53 (m, 4H), 7.49-7.13 (m, 39H), 7.08-7.06
(m, 2H), 7.01 (d, J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.56 (s,
1H), 5.44 (d, J=3.9 Hz, 1H), 5.36 (d, J=8.2 Hz, 1H), 5.33 (bs, 1H),
5.16-5.14 (m, 2H), 4.98 (d, J=3.8 Hz, 1H), 4.96 (d, J=5.1 Hz, 1H),
4.85 (d, J=11.4 Hz, 1H), 4.80-4.67 (m, 7H), 4.59 (d, J=7.9 Hz, 1H),
4.57 (d, J=3.8 Hz, 1H), 4.51-4.45 (m, 3H), 4.44-4.41 (m, 1H),
4.34-4.28 (m, 2H), 4.24-4.02 (m, 15H), 3.98-3.89 (m, 6H), 3.82-3.58
(m, 13H), 3.77 (s, 3H), 3.51 (t, J=9.9 Hz, 1H), 3.33-3.29 (m, 2H),
3.24 (dd, J=10.1, 3.7 Hz, 1H), 2.05, 2.00, 1.98 (s, 3H each).
.sup.13C NMR .delta. 170.51, 170.48, 166.84, 166.75, 166.55,
165.65, 165.47, 164.94, 155.24, 154.19, 150.14, 143.22, 143.07,
141.38, 141.35, 138.28, 137.88, 137.82, 137.61, 137.28, 137.21,
137.09, 133.64, 133.56, 129.90, 129.86, 129.65, 129.54, 129.07,
128.92, 128.78, 128.60, 128.48, 128.43, 128.31, 128.26, 128.21,
128.14, 128.09, 128.02, 127.88, 127.81, 127.72, 127.57, 127.46,
127.27, 127.22, 125.34, 125.05, 124.85, 120.17, 120.15, 118.30,
114.56, 100.88, 98.88, 98.81, 97.99, 97.83, 97.52, 82.80, 78.71,
78.38, 77.94, 77.69, 75.45, 75.30, 75.20, 75.14, 75.02, 74.68,
74.64, 74.56, 74.44, 74.19, 73.85, 73.35, 72.46, 72.38, 70.32,
70.20, 70.02, 69.77, 68.83, 68.09, 67.08, 65.60, 64.87, 64.75,
63.90, 63.41, 63.34, 63.14, 62.11, 62.07, 61.97, 55.70, 46.77,
40.66, 40.45, 40.43, 20.80, 20.72, 20.65.
[0606] Synthesis of 115
[0607] Compound 115 is prepared from compound 114 following general
procedure D.
[0608] .sup.1H NMR; (CDCl.sub.3) .delta. 8.13-8.07 (m, 6H),
7.58-7.41 (m, 10 H), 7.34-7.12 (m, 29H), 7.01 (d, J=9.1 Hz, 2H),
6.83 (d, J=9.1 Hz, 2H), 5.56 (s, 1H), 5.43 (d, J=3.9 Hz, 1H),
5.36-5.32 (m, 2H), 5.15 (t, J=4 Hz, 1H), 5.11 (d, J=3.7 Hz, 1H),
4.98 (d, J=3.9 Hz, 1H), 4.96 (d, J=5.3 Hz, 1H), 4.84 (d, J=11.3 Hz,
1H), 4.79-4.66 (m, 7H), 4.60-4.45 (m, 6H), 4.41-4.37 (m, 2H), 4.31
(dd, J=12.2, 3.6 Hz, 1H), 4.25-4.06 (m, 10H), 3.98-3.89 (m, 5H),
3.80-3.68 (m, 11H), 3.77 (s, 3H), 3.62-3.55 (m, 4H), 3.50 (t, J=9
Hz, 1H), 3.37 (dt, J=9.4, 4.3 Hz, 1H), 3.31 (dd, J=9.8, 4.3 Hz,
1H), 3.26-3.20 (m, 2H), 2.82 (d, J=4.2 Hz, 1H), 2.04, 2.03, 1.97
(s, 3H each). .sup.13C NMR .delta. 171.69, 170.67, 170.49, 166.99,
166.68, 166.53, 165.63, 165.52, 164.92, 155.21, 150.12, 138.25,
137.82, 137.68, 137.59, 137.25, 133.62, 133.55, 133.49, 129.86,
129.61, 129.51, 129.05, 128.76, 128.69, 128.59, 128.39, 128.26,
128.11, 128.06, 127.96, 127.81, 127.78, 127.55, 127.42, 127.18,
125.32, 118.28, 114.54, 100.85, 99.01, 98.79, 97.92, 97.82, 97.49,
82.76, 79.71, 78:57, 78.36, 77.64, 75.43, 75.33, 75.19, 74.75,
74.64, 74.24, 74.17, 73.41, 72.44, 72.36, 72.31, 71.27, 70.56,
70.43, 70.26, 69.99, 68.06, 67.72, 65.57, 64.82, 64.72, 63.75,
63.31, 63.09, 62.80, 62.08, 62.01, 55.69, 40.67, 40.44, 40.38,
20.79, 20.72.
[0609] Synthesis of 116
[0610] Compound 116 is prepared from compound 115 and 23 following
general procedure E.
[0611] .sup.1H NMR; (CDCl.sub.3) .delta. 8.13-8.05 (m, 8H),
7.58-7.52 (m, 4H), 7.49-7.41 (m, 10H), 7.37-7.11 (m, 43H), 7.01 (d,
J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.56 (s, 1H), 5.43-5.40 (m,
2H), 5.36-5.31 (m, 3H), 5.11 (t, J=4.1 Hz, 1H), 5.05 (d, J=3.8 Hz,
1H), 4.96 (d, J=11.7 Hz, 1H), 4.91 (d, J=10.5 Hz, 1H), 4.86 (br s,
2H), 4.83-4.74 (m, 6H), 4.70-4.62 (m, 7H), 4.59-4.55 (m, 3H),
4.48-4.45 (m, 1H), 4.30-4.12 (m, 16H), 4.06-3.56 (m, 31H), 3.77 (s,
3H), 3.53-3.46 (m, 2H), 3.31-3.19 (m, 4H), 2.01, 2.00, 1.97, 1.96
(s, 3H each). .sup.13C NMR .delta. 170.49, 170.43, 166.82, 166.65,
166.58, 166.53, 165.64, 165.44, 164.93, 155.23, 150.13, 138.24,
138.19, 137.80, 137.60, 137.52, 137.24, 133.68, 133.56, 129.85,
129.62, 129.37, 129.06, 128.78, 128.62, 128.57, 128.39, 128.36,
128.20, 128.12, 128.05, 127.98, 127.79, 127.56, 127.43, 127.21,
125.32, 118.29, 114.55, 100.95, 100.84, 98.81, 98.60, 98.00, 97.82,
97.51, 82.77, 80.13, 78.52, 78.36, 78.06, 77.76, 77.61, 75.57,
75.43, 75.35, 75.29, 75.23, 75.18, 74.93, 74.77, 74.63, 74.40,
74.17, 74.11, 73.49, 72.45, 72.37, 72.32, 70.35, 70.26, 70.14,
70.01, 69.89, 68.08, 67.53, 65.59, 65.03, 64.82, 64.73, 63.38,
63.32, 63.21, 62.40, 62.08, 61.91, 55.70, 40.59, 40.45, 40.38,
20.77, 20.70.
[0612] Synthesis of 117
[0613] Compound 117 is prepared from compound 116 following general
procedure F.
[0614] .sup.1H NMR; (CDCl.sub.3) .delta. 8.14-8.05 (m, 8H),
7.59-7.56 (m, 2H), 7.53-7.49 (m, 2H), 7.47-7.41 (m, 10H), 7.37-7.12
(m, 43H), 7.01 (d, J=9.1 Hz, 2H), 6.82 (d, J=9.1 Hz, 2H), 5.59 (s,
1H), 5.54 (d, J=4.0 Hz, 1H), 5.51 (d, J=3.9 Hz, 1H), 5.34-5.29 (m,
3H), 5.09 (t, J=2.4 Hz, 1H), 5.01 (s, 1H), 4.98 (d, J=11.7 Hz, 1H),
4.86 (br s, 3H), 4.83-4.64 (m,11H), 4.58-4.53 (m, 4H), 4.46 (d,
J=2.8 Hz, 1H), 4.45 (d, J=2.8 Hz, 1H), 4.38 (t, J=6.8 Hz, 1H), 4.29
(d, J=12.2 Hz, 1H), 4.22-3.62 (m, 29H), 3.77 (s, 3H), 3.56-3.44 (m,
9H), 3.35-3.24 (m, 6H), 3.16-3.06 (m, 3H), 2.02, 2.00, 1.96, 1.95
(s, 3H each). .sup.13C NMR .delta. 170.57, 170.41, 165.71, 164.95,
155.16, 150.33, 138.12, 137.77, 137.58, 137.47, 137.36, 137.31,
137.25, 133.60, 133.44, 129.84, 129.63, 129.08, 128.74, 128.69,
128.56, 128.53, 128.45, 128.35, 128.24, 128.06, 128.02, 127.87,
127.81, 127.76, 127.64, 126.84, 126.79, 125.31, 118.22, 114.65,
101.27, 101.21, 98.58, 98.10, 97.84, 97.69, 97.60, 97.36, 83.62,
80.25, 78.95, 78.47, 78.32, 78.13, 78.01, 77.75, 75.54, 75.13,
75.04, 74.88, 74.80, 74.35, 74.27, 73.81, 73.39, 73.11, 72.99,
72.74, 72.16, 72.06, 70.08, 69.93, 69.60, 68.80, 68.33, 67.56,
63.49, 63.40, 63.22, 62.61, 62.39, 62.26, 62.10, 61.75, 61.63,
61.30, 61.02, 55.66, 20.82, 20.73, 20.64.
[0615] Synthesis of 118
[0616] Compound 118 is prepared from compound 117 following general
procedure G2.
[0617] .sup.1H NMR; (CDCl.sub.3) .delta. 8.12-8.05 (m, 8H),
7.60-7.42 (m, 12H), 7.38-7.09 (m, 45H), 6.98 (d, J=9.1 Hz, 2H),
6.80 (d, J=9.1 Hz, 2H), 5.63 (s, 1H), 5.48 (d, J=6.4 Hz, 1H),
5.45-5.37 (m, 4H), 5.29 (t, J=2.6 Hz, 1H), 5.15 (t, J=6.9 Hz, 1H),
5.03 (d, J=10.9 Hz, 1H), 4.95-4.91 (m, 3H), 4.88 (br s, 2H),
4.83-4.55 (m, 15H), 4.44 (d, J=10.9 Hz, 1H), 4.31-3.42 (m, 31H),
3.76, 3.52, 3.51, 3.50 (s, 3H each), 3.36-3.31 (m, 2H), 3.27 (dd,
J=10.3, 3.8 Hz, 1H), 3.23 (dd, J=10.3, 3.6 Hz, 1H), 3.13 (dd,
J=10.3, 3.6 Hz, 1H), 2.97, 2.12, 2.03, 2.01, 2.01 (s, 3H each).
.sup.13C NMR .delta. 170.87, 170.60, 170.43, 169.72, 169.16,
167.69, 167.49, 165.63, 165.08, 164.69, 155.27, 150.54, 137.93,
137.56, 137.39, 137.18, 133.92, 133.63, 129.94, 129.83, 129.73,
129.35, 129.03, 128.78, 128.54, 128.31, 128.17, 128.06, 127.99,
127.91, 127.76, 127.59, 127.33, 118.10, 114.62, 101.18, 98.68,
98.27, 97.69, 97.24, 82.80, 82.47, 80.22, 78.04, 77.82, 77.72,
77.61, 75.95, 75.58, 75.31, 75.20, 75.01, 74.93, 74.82, 74.68,
74.53, 73.65, 73.45, 72.67, 72.16, 71.56, 69.86, 69.54, 69.43,
69.30, 68.50, 68.27, 63.41, 63.01, 62.88, 62.18, 61.54, 55.67,
52.69, 52.08, 51.96, 51.86, 20.81.
[0618] Synthesis of 119
[0619] Compound 119 is prepared from compound 118 following general
procedure H.
[0620] .sup.1H NMR; (CDCl.sub.3) .delta. 8.07-7.98 (m, 8H),
7.57-7.54 (m, 4H), 7.47-7.43 (m, 8H), 7.35-7.02 (m, 45H), 6.96 (d,
J=9.1 Hz, 2H), 6.78 (d, J=9.1 Hz, 2H), 5.71-5.67 (m, 2H), 5.61-5.57
(m, 2H), 5.44-5.37 (m, 3H), 5,32 (m, 2H), 5.19-5.16 (m, 2H), 5.02
(d, J=3.3 Hz, 1H), 4.96-4.77 (m, 10H), 4.69-4.47 (m, 14H),
4.40-3.91 (m, 20H), 3.84-3.27 (m, 14H), 3.75, 3.52, 3.49, 3.39,
3.29, 2.10, 2.01, 1.98, 1.94, 1.40, 1.39, 1.38, 1.30 (s, 3H each).
.sup.13C NMR .delta. 171.11, 170.68, 170.62, 170.27, 170.08,
169.98, 169.26, 168.92, 167.29, 167.23, 165.56, 165.37, 164.72,
155.38, 150.46, 138.70, 138.57, 138.12, 137.70, 137.04, 136.85,
136.37, 133.93, 133.86, 129.78, 129.73, 129.04, 128.98, 128.93,
128.88, 128.83, 128.71, 128.62, 128.57, 128.52, 128.30, 128.24,
128.18, 128.09, 128.06, 128.00, 127.90, 127.77, 127.67, 127.61,
127.32, 127.27, 118.11, 114.64, 101.35, 101.29, 99.60, 98.87,
98.11, 97.85, 97.77, 97.70, 81.62, 81.40, 80.51, 77.98, 77.92,
77.47, 75.45, 75.27, 75.18, 74.95, 74.85, 74.75, 74.67, 73.75,
73.59, 73.52, 73.32, 73.22, 72.66, 71.94, 70.97, 70.71, 70.46,
69.84, 69.13, 68.61, 66.97, 62.18, 61.60, 61.36, 55.65, 52.78,
52.52, 52.47, 52.17, 52.09, 51.85, 51.78, 51.68, 22.74, 22.70,
22.61, 20.96, 20.82, 20.79, 20.73.
[0621] Synthesis of 120
[0622] Compound 120 is prepared from compound 119 following general
procedure I.
[0623] .sup.1H NMR; (CDCl.sub.3) .delta. 8.05-8.02 (m, 4H), 7.98
(d, J=7.3 Hz, 2H), 7.92 (d, J=7.3 Hz, 2H), 7.58-7.52 (m, 4H),
7.47-7.41 (m, 8H), 7.34-7.01 (m, 45H), 6.93 (d, J=9.1 Hz, 2H), 6.75
(d, J=9.1 Hz, 2H), 5.72 (d, J=9.4 Hz, 1H), 5.64 (d, J=8.9 Hz, 1H),
5.57-5.54 (m, 2H), 5.46-5.40 (m, 3H), 5.35-5.33 (m, 2H), 5.20 (t,
J=4.2 Hz, 1H), 5.13-5.11 (m, 2H), 4.99-4.48 (m, 24H), 4.34 (d,
J=12.2 Hz, 1H), 4.25-4.20 (m, 2H), 4.16-3.24 (m, 31H), 3.73, 3.52,
3.44, 3.31, 3.27, 1.46, 1.43, 1.42, 1.34 (s, 3H each). .sup.13C
NMR; .delta. 170.49, 170.16, 169.91, 169.06, 167.97, 167.87,
165.48, 164.79, 155.34, 150.47, 138.91, 138.59, 138.30, 137.94,
137.07, 136.99, 136.52, 133.98, 133.92, 133.84, 133.77, 129.74,
129.66, 129.54, 129.01, 128.86, 128.61, 128.51, 128.28, 128.18,
128.05, 127.99, 127.85, 127.76, 127.68, 127.50, 127.30, 118.04,
114.63, 100.81, 100.62, 99.07, 98.52, 98.42, 98.15, 97.61, 97.24,
81.45, 81.34, 80.13, 77.74, 77.56, 76.49, 75.50, 75.24, 74.98,
74.89, 74.67, 74.40, 73.76, 73.66, 73.33, 72.98, 72.85, 72.79,
72.11, 71.89, 71.47, 70.71, 68.98, 68.72, 61.62, 60.50, 60.39,
59.98, 55.64, 52.74, 52.65, 52.36, 52.25, 51.97, 51.77, 22.68,
22.65.
[0624] Synthesis of 121
[0625] Compound 121 is prepared from compound 120 following general
procedure J.
[0626] .sup.1H NMR; (CD.sub.3OD) .delta. 8.24-8.22 (m,
2H),8.18-8.16 (m, 2H), 8.11-8.09 (m, 2H), 7.98-7.96 (m, 2H),
7.68-7.45 (m, 12H), 7.36-7.08 (m, 41H), 6.98-6.96 (m, 4H), 6.92 (d,
J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.57 (d, J=4.5 Hz, 1H), 5.44
(d, J=3.0 Hz, 1H), 5.39-5.21 (m, 8H), 5.02-5.00 (m, 2H), 4.96 (d,
J=3.5 Hz, 1H), 4.90-4.52 (m, 24H), 4.47-4.43 (m, 2H), 4.37-4.08 (m,
15H), 4.03-3.93 (m, 6H), 3.76-3.65 (m, 3H), 3.75 (s, 3H), 3.62-3.55
(m, 4H), 3.57 (s, 3H), 3.46-3.41 (m, 2H), 3.24, 2.98, 1.86, 1.84,
1.59, 1.53 (s, 3H each). .sup.13C NMR; .delta. 173.58, 173.35,
171.09, 170.61, 170.40, 170.26, 167.34, 167.12, 166.58, 157.06,
152.01, 140.26, 140.16, 140.07, 139.92, 139.87, 139.01, 138.71,
138.47, 135.13, 134.99, 134.85, 134.66, 131.24, 131.16, 130.93,
130.86, 130.66, 130.52, 130.22, 129.97, 129.52, 129.37, 129.32,
129.25, 129.21, 129.16, 129.07, 128.99, 128.85, 128.68, 128.61,
128.42, 128.18, 119.64, 115.74, 101.96, 101.68, 99.95, 98.97,
98.71, 97.33, 84.05, 83.94, 81.71, 79.93, 79.41, 79.12, 78.98,
78.76, 78.17, 76.52, 76.23, 76.10, 75.85, 75.69, 75.41, 75.07,
75.01, 74.48, 74.07, 73.82, 72.54, 71.77, 71.59, 71.48, 70.95,
70.09, 66.85, 66.08, 65.74, 56.19, 54.09, 53.78, 53.49, 53.36,
53.10, 53.02, 52.58, 22.94, 22.85, 22.65.
[0627] Synthesis of 122
[0628] Compound 122 is prepared from compound 121 following general
procedure K.
[0629] .sup.1H NMR; (CD.sub.3OD) .delta. 7.43-7.19 (m, 45H), 7.07
(d, J=9.1 Hz, 2H), 6.84 (d, J=9.1 Hz, 2H), 5.47 (d, J=1.2 Hz, 1H),
5.41 (d, J=3.8 Hz, 1H), 5.39 (d, J=3.8 Hz, 1H), 5.22 (d, J=2.5 Hz,
1H), 5.09-4.54 (m, 22H), 4.44 (d, J=11.8 Hz, 1H), 4.37-3.68 (m,
35H), 3.74 (s, 3H), 3.63-3.52 (m, 4H), 1.86, 1.78, 1.77, 1.70 (s,
3H each). .sup.13C NMR; .delta. 177.80, 176.22, 176.04, 175.71,
175.63, 173.33, 173.16, 173.08, 156.51, 152.45, 140.40, 140.28,
140.15, 139.88, 139.55, 139.37, 129.48, 129.43, 129.37, 129.31,
129.25, 129.05, 128.96, 128.86, 128.79, 128.56, 128.45, 119.34,
115.66, 104.07, 103.96, 102.07, 101.71, 98.54, 98.46, 97.09, 96.08,
85.87, 81.97, 81.07, 80.82, 80.48, 79.38, 78.42, 77.64, 77.51,
76.65, 76.30, 75.94, 75.58, 75.13, 75.00, 74.70, 74.32, 73.12,
72.93, 72.32, 71.32, 71.18, 71.09, 70.55, 70.11, 68.49, 67.49,
67.07, 56.13, 54.45, 53.64, 53.59, 22.97, 22.52.
[0630] Synthesis of 123
[0631] Compound 123 is prepared from compound 122 following general
procedure L.
[0632] .sup.1H NMR; (D.sub.2O) .delta. 6.99 (d, J=8.3 Hz, 2H), 6.83
(d, J=8.3 Hz, 2H), 5.30 (s, 1H), 5.26 (s, 1H), 5.22 (s, 1H),
5.02-4.99 (m, 2H), 4.82 (s, 1H), 4.61-4.54 (br s, HOD and 2H),
4.43-4.41 (m, 2H), 4.30-4.28 (m, 2H), 4.20-4.17 (m, 2H), 4.08-4.00
(m, 5H), 3.95-3.72 (m, 13H), 3.67-3.54 (m, 16H), 3.41 (t, J=9.4 Hz,
1H), 3.20-3.17 (m, 2H), 1.89, 1.85 (s, 6H each). .sup.13C NMR
.delta. 175.07, 174.76, 174.53, 174.49, 174.43, 174.38, 154.82,
150.34, 119.38, 115.19, 102.04, 101.96, 101.90, 100.72, 97.11,
97.06, 94.49, 94.35, 77.39, 77.16, 76.90, 76.63, 76.57, 76.28,
76.19, 74.34, 73.84, 73.56, 70.66, 70.10, 69.57, 69.40, 69.28,
69.23, 69.16, 68.96, 68.88, 68.08, 66.41, 66.23, 65.92, 55.93,
53.76, 53.60, 53.05, 22.06, 22.02.
[0633] Synthesis of 124
[0634] Compound 124 is prepared from compounds 24 and 27 following
general procedure E.
[0635] .sup.1H NMR; (CDCl.sub.3) .delta. 8.11-8.05 (m, 4H), 7.76
(d, J=7.6 Hz, 2H), 7.60-7.54 (m, 3H), 7.50-7.13 (m, 27H), 7.09-7.07
(m, 2H), 6.89 (d, J=9.1 Hz, 2H), 6.76 (d, J=9.1 Hz, 2H), 5.55-5.52
(m, 2H), 5.17 (t, J=3.7 Hz, 1H), 5.14 (d, J=3.4 Hz, 1H), 5.08 (d,
J=7.1 Hz, 1H), 4.97 (d, J=10.4 Hz, 1H), 4.85 (d, J=11.3 Hz, 1H),
4.81-4.74 (m, 5H), 4.70 (d, J=10.4 Hz, 1H), 4.61 (dd, J=11.5, 2.6
Hz, 1H), 4.52-4.48 (m, 2H), 4.44-4.41 (m, 1H), 4.36-4.28 (m, 3H),
4.26-4.21 (m, 4H), 4.18 (t, J=6.8 Hz, 1H), 4.12-3.94 (m, 10H),
3.90-3.80 (m, 4H), 3.74 (s, 3H), 3.72-3.68 (m, 2H), 3.34-3.31 (m,
2H), 2.06, 2.00 (s, 3H each). .sup.13C NMR; .delta. 170.60, 170.50,
166.87, 166.83, 165.45, 165.11, 155.72, 154.19, 150.99, 143.20,
143.05, 141.34, 137.85, 137.34, 137.21, 137.06, 133.59, 133.49,
129.83, 129.50, 129.46, 129.06, 128.76, 128.61, 128.46, 128.42,
128.31, 128.25, 128.18, 128.01, 127.89, 127.81, 127.67, 127.27,
125.32, 125.04, 124.85, 120.14, 118.77, 114.57, 100.13, 98.86,
97.99, 97.79, 82.77, 78.64, 77.89, 75.41, 75.28, 75.18, 75.03,
74.56, 74.22, 74.13, 73.65, 73.43, 72.54, 70.32, 69.97, 68.81,
67.30, 64.65, 63.84, 63.36, 63.11, 62.25, 61.96, 55.66, 46.76,
40.66, 40.55, 20.79, 20.64.
[0636] Synthesis of 125
[0637] Compound 125 is prepared from compound 124 following general
procedure D.
[0638] .sup.1H NMR; (CDCl.sub.3) .delta. 8.11-8.09 (m, 2H),
8.07-8.05 (m, 2H), 7.57 (t, J=7.4 Hz, 1H), 7.52 (t, J=7.4 Hz, 1H),
7.46-7.43 (m, 4H), 7.37-7.19 (m, 20H), 6.88 (d, J=9.1 Hz, 2H), 6.76
(d, J=9.1 Hz, 2H), 5.55-5.52 (m, 2H), 5.18 (t, J=4.2 Hz, 1H), 5.12
(d, J=3.9 Hz, 1H), 5.07 (d, J=7.1 Hz, 1H), 4.97 (d, J=10.3 Hz, 1H),
4.84 (d, J=11.3 Hz, 1H), 4.80-4.75 (m, 4H), 4.69 (d, J=10.3 Hz,
1H), 4.61-4.47 (m, 4H), 4.43 (d, J=10.8 Hz, 1H), 4.39-4.34 (m, 2H),
4.29 (dd, J=11.8, 5.8 Hz, 1H), 4.20 (d, J=11.6 Hz, 1H), 4.16-4.05
(m, 4H), 4.00 (t, J=8.7 Hz, 1H), 3.94-3.93 (m, 4H), 3.88-3.79 (m,
4H), 3.75-3.71 (m, 2H), 3.74 (s, 3H), 3.58 (dd, J=10.0, 8.8 Hz,
1H), 3.40-3.36 (m, 1H), 3.32 (dd, J=9.7, 4.0 Hz, 1H), 3.22 (dd,
J=10.1, 3.7 Hz, 1H), 2.82 (d, J=4.2 Hz, 1H), 2.06, 2.04 (s, 3H
each). .sup.13C NMR; .delta. 171.69, 170.74, 167.01, 166.75,
165.49, 165.08, 155.69, 150.98, 137.87, 137.68, 137.34, 137.26,
133.53, 133.47, 129.83, 129.49, 129.44, 128.70, 128.59, 128.41,
128.33, 128.26, 128.10, 127.79, 127.66, 118.75, 114.55, 100.10,
99.00, 97,93, 97.78, 82.74, 79.68, 78.51, 75.61, 75.30, 75.20,
74.73, 74.56, 74.21, 73.62, 73.50, 72.52, 71.26, 70.62, 70.56,
70.29, 67.94, 64.60, 63.69, 63.27, 63.06, 62.81, 62.20, 55.65,
40.67, 40.52, 20.78, 20.72.
[0639] Synthesis of 126
[0640] Compound 126 is prepared from compounds 125 and 22 following
general procedure E.
[0641] .sup.1H NMR; (CDCl.sub.3) .delta. 8.10-8.05 (m, 6H),
7.76-7.73 (m, 2H), 7.59-7.53 (m, 5H), 7.47-7.42 (m, 6H), 7.39-7.36
(m, 2H), 7.31-7.13 (m, 32H), 6.88 (d, J=9.1 Hz, 2H), 6.76 (d, J=9.1
Hz, 2H), 5.54-5.51 (m, 2H), 5.47 (d, J=3.8 Hz, 1H), 5.36 (t, J=8.1
Hz, 1H), 5.14 (t, J=4.4 Hz, 1H), 5.07-5.06 (m, 2H), 4.91 (d, J=10.5
Hz, 1H), 4.88-4.84 (m, 2H), 4.80-4.62 (m, 10H), 4.57 (dd, J=11.7,
2.5 Hz, 1H), 4.47 (dd, J=10.4, 6.7 Hz, 1H), 4.33-4.13 (m, 11H),
4.08-3.75 (m, 23H), 3.73 (s, 3H), 3.67-3.59 (m, 4H), 3.35 (dd,
J=10.3, 3.9 Hz, 1H), 3.29 (dd, J=9.6, 4.0 Hz, 1H), 3.23 (dd, J=9.7,
3.8 Hz, 1H), 2.03, 2.02, 1.97 (s, 3H each). .sup.13C NMR; .delta.
170.58, 170.44, 166.87, 166.74, 166.62, 165.43, 165.08, 164.93,
155.70, 154.12, 150.99, 143.25, 143.01, 141.34, 138.23, 137.86,
137.36, 137.25, 137.14, 137.09, 133.74, 133.61, 133.47, 129.83,
129.52, 129.32, 129.06, 128.82, 128.75, 128.60, 128.41, 128.32,
128.21, 127.98, 127.93, 127.80, 127.68, 127.57, 127.24, 125.33,
125.11, 124.91, 120.13, 118.77, 114.56, 100.94, 100.11, 98.62,
97.82, 82.74, 82.68, 78.46, 78.06, 77.76, 77.48, 75.67, 75.32,
75.20, 74.97, 74.77, 74.58, 74.22, 74.08, 73.58, 72.54, 72.32,
70.40, 70.31, 69.88, 68.77, 67.71, 64.96, 64.59, 63.36, 63.26,
63.17, 62.61, 62.13, 61.94, 61.81, 55.65, 46.71, 40.60, 40.53,
40.43, 20.77, 20.70.
[0642] Synthesis of 127
[0643] Compound 127 is prepared from compound 126 following general
procedure D.
[0644] .sup.1H NMR; (CDCl.sub.3) .delta. 8.10-8.04 (m, 6H),
7.58-7.53 (m, 3H), 7.47-7.41 (m, 6H), 7.40-7.35 (m, 4H), 7.32-7.13
(m, 26H), 6.88 (d, J=9.1 Hz, 2H), 6.76 (d, J=9.1 Hz, 2H), 5.54-5.51
(m, 2H), 5.44 (d, J=3.9 Hz, 1H), 5.35 (t, J=8.2 Hz, 1H), 5.14 (t,
J=4.4 Hz, 1H), 5.07-5.05 (m, 2H), 4.92-4.83 (m, 4H), 4.80-4.76 (m,
4H), 4.73-4.64 (m, 5H), 4.57 (dd, J=11.8, 2.6 Hz, 1H), 4.51 (dd,
J=12.5, 3.6 Hz, 1H), 4.33-4.11 (m, 9H), 4.08-3.92 (m, 9H),
3.87-3.72 (m, 10H), 3.73 (s, 3H), 3.66-3.58 (m, 6H), 3.41 (dt,
J=9.5, 3.9 Hz, 1H), 3.29 (dd, J=9.6, 3.9 Hz, 1H), 3.24 (t, J=4.5
Hz, 1H), 3.22 (t, J=4.1 Hz, 1H), 2.92 (d, J=4.0 Hz, 1H), 2.07,
2.03, 1.96 (s, 3H each). .sup.13C NMR; .delta. 171.88, 170.57,
170.47, 166.90, 166.73, 166.61, 165.42, 165.08, 164.95, 155.70,
150.99, 138.27, 137.85, 137.78, 137.35, 137.25, 137.21, 133.71,
133.60, 133.46, 129.82, 129.52, 129.32, 129.05, 128.80, 128.74,
128.67, 128.59, 128.39, 128.31, 128.24, 128.20, 128.04, 127.96,
127.79, 127.72, 127.67, 127.60, 127.54, 127.16, 125.32, 118.77,
114.56, 100.92, 100.11, 98.61, 98.22, 97.83, 97.78, 82.81, 82.74,
79.16, 78.45, 78.05, 77.78, 75.66, 75.42, 75.25, 75.02, 74.96,
74.72, 74.57, 74.21, 74.15, 73.62, 73.57, 72.54, 72.44, 71.36,
70.62, 70.30, 69.88, 67.71, 64.92, 64.58, 63.34, 63.25, 63.15,
62.72, 62.66, 62.13, 61.95, 55.65, 40.59, 40.52, 40.43, 20.78,
20.70.
[0645] Synthesis of 128
[0646] Compound 128 is prepared from compound 127 and 25 following
general procedure E.
[0647] .sup.1H NMR; (CDCl.sub.3) .delta. 8.09-8.04 (m, 8H),
7.58-7.54 (m, 3H), 7.47-7.43 (m, 7H), 7.39-7.12 (m, 47H), 6.88 (d,
J=9.1 Hz, 2H), 6.76 (d, J=9.1 Hz, 2H), 5.54-5.50 (m, 2H), 5.41 (d,
J=3.9 Hz, 1H), 5.33 (t, J=8.2 Hz, 1H), 5.14-5.11 (m, 3H), 5.07-5.05
(m, 2H), 4.97 (d, J=11.3 Hz, 1H), 4.91 (d, J=10.4 Hz, 1H),
4.84-4.81 (m, 2H), 4.79-4.62 (m, 10H), 4.57 (dd, J=11.7, 2.5 Hz,
1H), 4.53-4.47 (m, 3H), 4.38 (m, 1H), 4.33-3.91 (m, 22H), 3.86-3.59
(m, 15H), 3.73 (s, 3H), 3.42 (t, J=9.5 Hz, 1H), 3.31-3.21 (m, 4H),
2.03, 2.02, 1.98, 1.96 (s, 3H each). .sup.13C NMR; .delta. 170.57,
170.51, 170.41, 166.86, 166.70, 166.61, 165.48, 165.40, 165.07,
164.92, 155.69, 150.98, 138.22, 137.84, 137.77, 137.45, 137.35,
137.24, 133.60, 133.45, 129.84, 129.51, 129.30, 129.05, 128.78,
128.74, 128.70, 128.62, 128.48, 128.39, 128.29, 128.24, 128.19,
128.11, 128.06, 127.97, 127.84, 127.79, 127.72, 127.67, 127.53,
127.26, 125.32, 118.77, 114.54, 100.90, 100.10, 98.80, 98.61,
97.99, 97.77, 82.78, 82.73, 80.41, 78.64, 78.44, 77.99, 77.72,
77.62, 75.69, 75.34, 75.22, 75.16, 74.92, 74.74, 74.67, 74.58,
74.21, 74.15, 73.60, 73.43, 72.53, 72.36, 70.31, 70.11, 69.88,
67.76, 67.52, 64.76, 64.57, 63.80, 63.66, 63.32, 63.23, 63.12,
62.60, 62.10, 61.98, 61.88, 55.64, 40.67, 40.59, 40.51, 40.41,
20.76, 20.72.
[0648] Synthesis of 129
[0649] Compound 129 is prepared from compound 128 following general
procedure F.
[0650] .sup.1H NMR; (CDCl.sub.3) .delta. 8.11-8.04 (m, 8H),
7.59-7.50 (m, 3H), 7.47-7.12 (m, 54H), 6.89 (d, J=9.1 Hz, 2H), 6.76
(d, J=9.1 Hz, 2H), 5.63 (d, J=3.9 Hz, 1H), 5.56-5.53 (m, 2H), 5.32
(dd, J=8.8, 8.3 Hz, 1H), 5.14-5.10 (m, 3H), 5.05-5.04 (m, 2H),
4.88-4.84 (m, 2H), 4.82-4.62 (m, 14H), 4.51-4.46 (m, 3H), 4.33-4.03
(m, 16H), 3.97-3.65 (m, 18H), 3.73 (s, 3H), 3.58-3.50 (m, 6H),
3.41-3.37 (m, 2H), 3.33-3.26 (m, 6H), 3.19-3.11 (m, 2H), 2.03 (s,
6H), 1.99, 1.97 (s, 3H each). .sup.13C NMR; .delta. 170.57, 170.52,
170.48, 170.42, 165.76, 165.71, 165.09, 164.95, 155.64, 151.12,
138.12, 137.88, 137.68, 137.59, 137.53, 137.48, 137.38, 137.35,
137.23, 133.63, 133.48, 133.39, 133.32, 129.83, 129.60, 129.06,
128.76, 128.68, 128.62, 128.56, 128.51, 128.41, 128.25, 128.13,
128.08, 127.96, 127.92, 127.84, 127.79, 127.75, 127.66, 126.89,
125.32, 118.52, 114.65, 101.21, 100.52, 98.21, 97.90, 97.41, 97.35,
83.64, 83.33, 80.64, 78.98, 78.74, 78.30, 77.96, 77.80, 75.25,
75.17, 75.10, 75.04, 74.89, 74.48, 74.43, 74.37, 74.32, 74.07,
73.95, 73.78, 73.44, 73.32, 73.16, 72.87, 72.51, 70.19, 70.07,
70.03, 69.65, 69.18, 67.97, 67.86, 63.83, 63.52, 63.41, 62.80,
62.62, 62.29, 62.14, 61.94, 61.71, 61.27, 61.04, 55.64, 20.83,
20.80, 20.75.
[0651] Synthesis of 130
[0652] Compound 130 is prepared from compound 129 following general
procedure G2.
[0653] .sup.1H NMR; (CDCl.sub.3) .delta. 8.12-8.09 (m, 4H),
8.06-8.01 (m, 4H), 7.57-7.53 (m, 3H), 7.49-7.13 (m, 52H), 7.09-7.07
(m, 2H), 6.84 (d, J=9.1 Hz, 2H), 6.73 (d, J=9.1 Hz, 2H), 5.51-5.47
(m, 4H), 5.44 (d, J=3.8 Hz, 1H), 5.38 (t, J=8.9 Hz, 1H), 5.17-5.15
(m, 2H), 5.06 (d, J=6.8 Hz, 1H), 5.01 (d, J=10.3 Hz, 1H), 4.98-4.96
(m, 2H), 4.93-4.90 (m, 2H), 4.80-4.62 (m, 13H), 4.56 (d, J=10.9 Hz,
1H), 4.48-4.38 (m, 3H), 4.34-4.29 (m, 4H), 4.26-4.01 (m, 12H),
3.96-3.63 (m, 11H), 3.72, 3.64 (s, 3H each), 3.55-3.48 (m, 2H),
3.49 (s, 3H), 3.44-3.41 (m, 2H), 3.31-3.19 (m, 4H), 3.21, 3.13,
2.12, 2.11, 2.00, 1.96 (s, 3H each). .sup.13C NMR; .delta. 170.83,
170.75, 170.47, 170.39, 169.70, 169.53, 168.10, 167.58, 165.23,
165.15, 164.99, 164.65, 155.68, 150.94, 137.97, 137.88, 137.84,
137.66, 137.54, 137.42, 137.30, 137.10, 133.93, 133.68, 133.51,
133.43, 129.97, 129.93, 129.79, 129.72, 129.45, 129.21, 129.05,
128.86, 128.80, 128.52, 128.48, 128.40, 128.33, 128.30, 128.16,
128.08, 127.96, 127.91, 127.88, 127.80, 127.58, 125.32, 118.73,
114.48, 101.18, 100.72, 99.17, 98.67, 98.24, 97.75, 97.39, 97.35,
82.71, 82.18, 80.00, 78.30, 77.93, 77.68, 77.60, 77.51, 76.05,
75.94, 75.54, 75.39, 75.19, 75.12, 75.02, 74.80, 74.75, 74.62,
74.52, 74.33, 74.21, 74.08, 73.80, 73.68, 72.06, 71.47, 70.75,
70.41, 70.11, 69.40, 63.51, 63.11, 62.90, 62.83, 62.29, 61.67,
61.53, 55.61, 52.19, 52.15, 52.08, 51.85, 20.81.
[0654] Synthesis of 131
[0655] Compound 131 is prepared from compound 130 following general
procedure H.
[0656] .sup.1H NMR; (CDCl.sub.3) .delta. 8.09-8.06 (m, 2H),
8.04-8.00 (m, 6H), 7.59-7.51 (m, 4H), 7.48-7.38 (m, 8H), 7.35-7.06
(m, 45H), 6.84 (d, J=9.1 Hz, 2H), 6.73 (d, J=9.1 Hz, 2H), 5.75 (d,
J=9.5 Hz, 1H), 5.68-5.60 (m, 3H), 5.52 (dd, J=8.5, 6.7 Hz, 1H),
5.42-5.37 (m, 2H), 5.33 (d, J=5.6 Hz, 1H), 5.19-5.17 (m, 2H),
5.06-5.05 (m, 2H), 5.01 (d, J=3.3 Hz, 1H), 4.94-4.75 (m, 8H),
4.69-4.47 (m, 13H), 4.42-3.92 (m, 23H), 3.86-3.46 (m, 11H), 3.71,
3.62, 3.53 (s, 3H each), 3.41-3.32 (m, 1H), 3.40, 3.34 (s, 3H
each), 2.11, 2.10, 1.99, 1.93, 1.50, 1.39, 1.38, 1.36 (s, 3H each).
.sup.13C NMR; .delta. 171.13, 171.04, 170.66, 170.57, 170.26,
170.17, 170.0, 169.97, 169.28, 169.20, 167.69, 167.28, 165.55,
165.42, 164.99, 164.70, 155.75, 150.86, 138.74, 138.59, 138.39,
138.05, 137.81, 136.89, 136.52, 136.34, 133.94, 133.84, 133.58,
129.75, 129.27, 129.04, 128.98, 128.93, 128.75, 128.62, 128.59,
128.47, 128.27, 128.18, 128.05, 127.79, 127.61, 127.56, 127.43,
127.35, 127.32, 125.31, 118.63, 114.52, 101.36, 100.90, 99.21,
99.05, 98.00, 97.91, 97.87, 97.73, 81.58, 80.95, 80.52, 78.08,
77.91, 75.58, 75.29, 74.97, 74.92, 74.84, 74.76, 74.50, 73.86,
73.77, 73.62, 73.49, 73.34, 72.87, 72.01, 71.10, 71.03, 70.61,
70.51, 70.37, 70.12, 69.82, 62.44, 61.69, 61.52, 55.61, 52.68,
52.57, 52.41, 52.09, 51.81, 22.95, 22.76, 22.58, 20.98, 20.76.
[0657] Synthesis of 132
[0658] Compound 132 is prepared from compound 131 following general
procedure I.
[0659] .sup.1H NMR; (CDCl.sub.3) .delta. 8.05-7.94 (m, 8H),
7.59-7.54 (m, 3H), 7.51-7.10 (m, 54H), 6.84 (d, J=9.1 Hz, 2H), 6.73
(d, J=9.1 Hz, 2H), 5.65 (d, J=9.3 Hz, 1H), 5.61 (d, J=9.3 Hz, 1H),
5.58-5.51 (m, 3H), 5.45 (d, J=4.3 Hz, 1H), 5.41 (t, J=8.2 Hz, 1H),
5.35 (d, J=5.2 Hz, 1H), 5.23-5.20 (m, 2H), 5.11 (d, J=3.4 Hz, 1H),
5.08-5.05 (m, 2H), 4.93-4.87 (m, 5H), 4.81-4.77 (m, 3H), 4.74-4.69
(m, 2H), 4.67-4.61 (m, 6H), 4.57-4.49 (m, 5H), 4.35-4.20 (m, 6H),
4.13-3.89 (m, 12H), 3.85 (t, J=8.2 Hz, 1H), 3.79 (d, J=10.2 Hz,
1H), 3.71 (s, 3H), 3.70-3.29 (m, 16H), 3.62, 3.56, 3.37, 3.27 (s,
3H each), 1.55, 1.45, 1.44, 1.41 (s, 3H each). .sup.13C NMR;
.delta. 170.45, 170.34, 169.94, 169.30, 168.98, 167.94, 165.56,
165.41, 164.99, 164.76, 155.75, 150.86, 138.98, 138.53, 138.46,
138.21, 138.09, 136.99, 136.52, 133.86, 133.58, 129.75, 129.64,
129.55, 129.26, 129.01, 128.90, 128.58, 128.51, 128.45, 128.25,
128.15, 128.03, 127.99, 127.92, 127.85, 127.76, 127.61, 127.42,
127.29, 118.61, 114.52, 100.87, 100.63, 98.87, 98.49, 97.81, 97.67,
97.47, 97.23, 81.38, 81.06, 79.84, 77.83, 76.59, 76.46, 75.37,
74.96, 74.75, 74.62, 74.49, 74.41, 74.03, 73.78, 73.34, 73.15,
73.02, 72.86, 72.77, 72.54, 71.91, 71.34, 70.91, 70.60, 70.02,
66.54, 66.36, 61.61, 60.83, 60.63, 60.39, 60.00, 55.61, 52.70,
52.58, 52.45, 52.22, 52.05, 51.78, 22.92, 22.70, 22.61.
[0660] Synthesis of 133
[0661] Compound 133 is prepared from compound 132 following general
procedure J.
[0662] .sup.1H NMR; (CD.sub.3OD) .delta. 8.28 (d, J=7.5 Hz, 2H),
8.11-8.05 (m, 6H), 7.65 (t, J=7.3 Hz, 1H), 7.58-7.07 (m, 52H),
7.02-7.00 (m, 2H), 6.93-6.91 (m, 2H), 6.87 (d, J=9.1 Hz, 2H), 6.77
(d, J=9.1 Hz, 2H), 5.58-5.52 (m, 3H), 5.43-5.38 (m, 3H), 5.27-5.21
(m, 4H), 5.08 (d, J=3.0 Hz, 1H), 5.00-4.97 (m, 2H), 4.93 (d, J=2.4
Hz, 1H), 4.82-4.10 (m, 38H), 4.00-3.95 (m, 3H), 3.87-3.84 (m, 2H),
3.69-3.30 (m, 21H), 2.90 (s, 3H), 1.86, 1.82, 1.66, 1.52 (s, 3H
each). .sup.13C NMR; .delta. 173.60, 173.43, 173.34, 173.25,
171.20, 170.46, 170.34, 167.38, 167.28, 166.95, 166.57, 164.90,
157.13, 152.29, 140.19, 140.02, 139.97, 139.79, 138.92, 138.64,
138.54, 138.38, 135.15, 134.81, 134.64, 134.53, 131.24, 130.96,
130.87, 130.81, 130.68, 130.22, 129.99, 129.80, 129.62, 129.54,
129.43, 129.33, 129.21, 129.10, 128.93, 128.81, 128.52, 128.42,
128.05, 119.10, 115.70, 101.82, 101.19, 99.17, 99.11, 99.00, 98.57,
97.03, 84.18, 83.38, 81.40, 79.97, 79.56, 79.17, 78.60, 76.27,
76.05, 75.59, 75.39, 75.27, 75.20, 74.95, 74.62, 74.36, 74.19,
73.37, 72.39, 72.16, 71.72, 71.58, 71.50, 71.24, 70.74, 70.39,
67.24, 66.20, 66.11, 65.87, 56.22, 54.40, 53.91, 53.60, 53.48,
53.24, 53.16, 53.08, 52.44, 49.94, 22.96, 22.91, 22.79.
[0663] Synthesis of 134
[0664] Compound 134 is prepared from compound 133 following general
procedure K.
[0665] .sup.1H NMR; (CD.sub.3OD) .delta. 7.43-7.11 (m, 45H), 7.04
(d, J=9.1 Hz, 2H), 6.84 (d, J=9.1 Hz, 2H), 5.41-5.39 (m, 2H),
5.25-5.23 (m, 2H), 5.09 (d, J=5.2 Hz, 1H), 5.07 (d, J=5.3 Hz, 1H),
4.98 (d, J=12.3 Hz, 1H), 4.91-4.69 (m, 11H), 4.66-4.54 (m, 8H),
4.45 (d, J=11.7 Hz, 1H), 4.39 (d, J=11.6 Hz, 1H), 4.36-4.24 (m,
5H), 4.18-3.61 (m, 32H), 3.75 (s, 3H), 3.51 (t, J=8.3 Hz, 1H),
1.82, 1.78, 1.77, 1.68 (s, 3H each). .sup.13C NMR; .delta. 175.64,
175.38, 174.72, 173.31, 173.17, 172.98, 156.89, 153.22, 140.30,
140.21, 140.10, 139.89, 139.86, 139.75, 139.71, 139.34, 139.24,
129.75, 129.64, 129.59, 129.52, 129.40, 129.35, 129.26, 129.02,
128.88, 128.76, 128.69, 128.64, 128.56, 128.48, 128.35, 128.16,
127.94, 119.55, 115.55, 103.92, 103.86, 102.02, 101.68, 98.78,
98.59, 98.04, 97.25, 86.18, 85.89, 82.13, 80.94, 80.68, 80.42,
79.09, 78.25, 77.72, 76.58, 76.23, 76.09, 75.81, 75.43, 75.22,
75.08, 74.55, 74.15, 73.13, 71.62, 71.38, 70.33, 69.77, 69.45,
68.96, 67.50, 67.20, 67.03, 56.14, 54.43, 54.24, 53.62, 49.91,
49.70, 22.97.
[0666] Synthesis of 135
[0667] Compound 135 is prepared from compound 134 following general
procedure L.
[0668] .sup.1H NMR; (D.sub.2O) .delta. 7.10 (d, J=7.5 Hz, 2H), 6.97
(d, J=7.5 Hz, 2H), 5.40-5.38 (m, 2H), 5.16 (s, 2H), 4.99-4.98 (m,
3H), 4.77 (br s, 2H), 4.57 (d, J=7.6 Hz, 1H), 4.44 (d, J=10.8 Hz,
1H), 4.35-4.33 (m, 3H), 4.23-4.18 (m, 4H), 4.08-4.06 (m, 3H),
4.03-4.01 (m, 2H), 3.97-3.89 (m, 8H), 3.85-3.69 (m, 14H), 3.81 (s,
3H), 3.59-3.54 (m, 2H), 3.34 (t, J=8.2 Hz, 1H), 2.05 (s, 6H), 2.02,
2.01 (s, 3H each). .sup.13C NMR; .delta. 175.04, 174.71, 174.53,
174.45, 154.87, 151.03, 118.39, 115.16, 102.02, 101.96, 101.86,
101.30, 97.20, 97.14, 94.53, 94.34, 77.30, 77.11, 76.81, 76.61,
76.43, 76.21, 74.22, 73.59, 73.38, 71.14, 70.23, 69.61, 69.39,
69.25, 68.98, 68.80, 66.54, 66.24, 65.93, 55.94, 53.78, 53.49,
53.05, 22.03.
Example 3
X-Ray Crystal Structure Analysis of Compounds 13, 14, 15, 16 and
27
[0669] Compound 13 is synthesised as described in Example 2 (GPA),
and crystallised from hot toluene by addition of petroleum ether.
X-Ray quality crystals are obtained by vapor diffusion of petroleum
ether into a solution of 13 in toluene.
[0670] Compound 14 is synthesised as described in Example 2 (GPA),
and crystallised from toluene by addition of petroleum ether. X-Ray
quality crystals are obtained by vapor diffusion of petroleum ether
into a solution of 14 in toluene.
[0671] Compound 15 is synthesised as described in Example 2 (GPA),
and crystallised from toluene by addition of petroleum ether. X-Ray
quality crystals are obtained by vapor diffusion of petroleum ether
into a solution of 15 in toluene.
[0672] Compound 16 is synthesised as described in Example 2 (GPA),
and crystallised from ethyl acetate by addition of petroleum
ether.
[0673] Compound 27 is synthesised as described in Example 2 (GPD),
and crystallises directly after elution from the chromatography
column. X-Ray quality crystals are obtained by vapor diffusion of
petroleum ether into a solution of 27 in toluene.
[0674] Crystals of compound 13 contain two enantiomerically
identical molecules which are aligned along the a axis and
separated by almost exactly by half a cell (shown in FIG. 1).
[0675] Data are collected on a Bruker APEXII diffractometer.
Computing details are as follows: data collection: CrystalClear
(Rigaku, 2005); cell refinement: FSProcess (Rigaku, 1998); data
reduction: FSProcess (Rigaku, 1998); program used to solve
structure: SHELXS97 (Sheldrick, 2008); program used to refine
structure: SHELXL97 (Sheldrick, 1997); molecular graphics: ORTEP in
WinGX (Farrugia, 1999).
TABLE-US-00001 TABLE 1 Crystal Data for Compound 13 Empirical
formula C59 H56 Cl N3 O16 Formula weight 1098.52 Temperature 123(2)
K Wavelength 1.54178 A Crystal system, space group Monoclinic, P 21
Unit cell dimensions a = 10.454(2) A alpha = 90 deg. b = 35.610(7)
A beta = 95.61(3) deg. c = 14.408(3) A gamma = 90 deg. Volume
5338.0(19) A{circumflex over ( )}3 Z, Calculated density 4, 1.367
Mg/m{circumflex over ( )}3 Absorption coefficient 1.271
mm{circumflex over ( )}-1 F(000) 2304 Crystal size 0.85 .times.
0.08 .times. ?0.04 mm Theta range for data collection 6.54 to 55.00
deg. Limiting indices -10 <= h <= 8, -37 <= k <= 37,
-15 <= l <= 15 Reflections collected/unique 35705/12625
[R(int) = 0.0554] Completeness to theta = 55.00 97.7% Absorption
correction Semi-empirical from equivalents Refinement method
Full-matrix least-squares on F{circumflex over ( )}2
Data/restraints/parameters 12625/3/1426 Goodness-of-fit on
F{circumflex over ( )}2 0.997 Final R indices [I > 2sigma(I)] R1
= 0.0575, wR2 = 0.1332 R indices (all data) R1 = 0.0840, wR2 =
0.1554 Absolute structure parameter 0.033(19) Largest diff. peak
and hole 0.318 and -0.301 e.A{circumflex over ( )}-3
TABLE-US-00002 TABLE 2 Crystal Data for Compound 14 Empirical
formula C51 H52 Cl N3 O14 Formula weight 966.41 Temperature 164(2)
K Wavelength 1.54178 .ANG. Crystal system, space group Monoclinic,
P 2.sub.1 Unit cell dimensions a = 9.2713(7) .ANG., alpha = 90 deg.
b = 17.4067(11) .ANG., beta = 97.449(7) deg. c = 15.0036(11) .ANG.,
gamma = 90 deg. Volume 2400.9(3) .ANG..sup.3 Z, Calculated density
2, 1.337 Mg/m.sup.3 Absorption coefficient 1.302 mm.sup.-1 F(000)
1016 Crystal size 0.85 .times. 0.45 .times. 0.14 mm Theta range for
data collection 7.01 to 60.01 deg. Limiting indices -10 <= h
<= 10, -19 <= k <= 14, -16 <= l <= 16 Reflections
collected/unique 15021/5678 [R(int) = 0.0556] Completeness to theta
= 60.01 99.2% Absorption correction Semi-empirical from equivalents
Max. and min. transmission 1.0, 0.521 Refinement method Full-matrix
least-squares on F.sup.2 Data/restraints/parameters 5678/3/558
Goodness-of-fit on F.sup.2 1.093 Final R indices [I > 2sigma(I)]
R1 = 0.0596, wR2 = 0.1600 R indices (all data) R1 = 0.0695, wR2 =
0.1714 Absolute structure parameter 0.02(3) (Flack, 1983)
Extinction coefficient 0.0023(4) Largest diff. peak and hole 0.311
and -0.316 e.A.sup.-3
TABLE-US-00003 TABLE 3 Crystal Data for Compound 15 Empirical
formula C52 H55 N3 O15 Formula weight 961.99 Temperature 118(2) K
Wavelength 0.71073 .ANG. Crystal system, space group Monoclinic, C2
Unit cell dimensions a = 38.3346(13) .ANG. alpha = 90 deg. b =
8.0744(3) .ANG. beta = 91.222(2) deg. c = 16.1659(6) .ANG. gamma =
90 deg. Volume 5002.7(3) .ANG..sup.3 Z, Calculated density 4, 1.277
Mg/m.sup.3 Absorption coefficient 0.094 mm.sup.-1 F(000) 2032
Crystal size 0.75 .times. 0.32 .times. 0.30 mm Theta range for data
collection 2.58 to 26.12 deg. Limiting indices -47 <= h <=
47, -9 <= k <= 9, -19 <= l <= 19 Reflections
collected/unique 51621/9796 [R(int) = 0.0350] Completeness to theta
= 26.12 98.8% Absorption correction Multi-scan (Blessing, 1995)
Max. and min. transmission 0.745 and 0.645 Refinement method
Full-matrix least-squares on F.sup.2 Data/restraints/parameters
9796/43/662 Goodness-of-fit on F.sup.2 1.079 Final R indices [I
> 2sigma(I)] R1 = 0.0484, wR2 = 0.1253 R indices (all data) R1 =
0.0525, wR2 = 0.1285 Absolute structure parameter 0.0(7)
(indeterminate) Largest diff. peak and hole 0.332 and -0.454
e.A.sup.-3
TABLE-US-00004 TABLE 4 Crystal Data for Compound 16 Empirical
formula C59 H56 Cl N3 O16 Formula weight 1098.52 Temperature 123(2)
K Wavelength 1.54178 A Crystal system, space group Monoclinic, P21
Unit cell dimensions a = 14.8343(11) A alpha = 90 deg. b =
8.4771(6) A beta = 91.780(7) deg. c = 21.8112(17) A gamma = 90 deg.
Volume 2741.5(4) A{circumflex over ( )}3 Z, Calculated density 2,
1.331 Mg/m{circumflex over ( )}3 Absorption coefficient 1.238
mm{circumflex over ( )}-1 F(000) 1152 Crystal size 0.6 .times. 0.05
.times. 0.02 mm Theta range for data collection 6.62 to 43.49 deg.
Limiting indices -13 <= h <= 13, -7 <= k <= 7, -19
<= l <= 19 Reflections collected/unique 19701/3962 [R(int) =
0.1013] Completeness to theta = 43.49 99.3% Absorption correction
Semi-empirical from equivalents Refinement method Full-matrix
least-squares on F{circumflex over ( )}2 Data/restraints/parameters
3962/55/666 Goodness-of-fit on F{circumflex over ( )}2 1.093 Final
R indices [I > 2sigma(I)] R1 = 0.0880, wR2 = 0.2135 R indices
(all data) R1 = 0.1479, wR2 = 0.2800 Absolute structure parameter
0.01(8) Extinction coefficient 0.0110(13) Largest diff. peak and
hole 0.289 and -0.247 e.A{circumflex over ( )}-3
TABLE-US-00005 TABLE 5 Crystal Data for Compound 27 Empirical
formula C44 H46 Cl N3 O14 Formula weight 876.29 Temperature 164(2)
K Wavelength 1.54178 .ANG. Crystal system, space group
Orthorhombic, P2.sub.12.sub.12.sub.1 Unit cell dimensions a =
8.1104(2) .ANG. alpha = 90 deg. b = 19.5548(6) .ANG. beta = 90 deg.
c = 27.2321(19) .ANG. gamma = 90 deg. Volume 4318.9(4) .ANG..sup.3
Z, Calculated density 4, 1.348 Mg/m.sup.3 Absorption coefficient
1.389 mm.sup.-1 F(000) 1840 Crystal size 1.0 .times. 0.13 .times.
0.11 mm Theta range for data collection 6.65 to 58.93 deg. Limiting
indices -6 <= h <= 8, -21 <= k <= 21, -30 <= l <=
30 Reflections collected/unique 34329/6132 [R(int) = 0.0454]
Completeness to theta = 58.93 99.3% Absorption correction
Semi-empirical from equivalents Max. and min. transmission 1.0,
0.717 Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 6132/10/527 Goodness-of-fit on F.sup.2
1.105 Final R indices [I > 2sigma(I)] R1 = 0.0638, wR2 = 0.1785
R indices (all data) R1 = 0.0827, wR2 = 0.1975 Absolute structure
parameter 0.17(4) (Flack, 1983) Largest diff. peak and hole 0.415
and -0.347 e.A.sup.-3
Example 4
Determination of BACE-1 Inhibition by In Vitro FRET Peptide
Cleavage Assay
[0676] The ability of compounds of the invention to inhibit BACE-1
cleavage of APP is assessed using a fluorescent resonance energy
transfer (FRET) peptide cleavage assay employing the FRET peptide
HiLyte 488-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys(QXL520)-OH (Anaspec,
Inc., CA, USA; Cat no. 60604-01). When intact, the amino terminal
fluorophore is quenched, but upon enzymatic cleavage the
fluorophore is released from quencher and fluoresces (520 nm).
Assays are performed in triplicate in 96 well black plates (20 mM
sodium acetate, 0.1% Triton-X-100, pH 4.5; 2.2 ng peptide per well
and 25 ng/well of recombinant human BACE-1 (R & D Systems Cat
no. 931-AS). The appropriate controls for enzyme activity
(substrate plus enzyme, and substrate only) are employed and plates
are incubated (1 h, 25.degree. C., with activity stopped with 2.5 M
sodium acetate). Compounds of the invention are added in the
concentration range from 100-0.0001 .mu.g/mL. Fluorescence
480ex/520em is measured on a Polarstar plate reader (BMG
LabTechnologies, UK) and data are analysed by plotting log.sub.10
concentration of compound against percent inhibition and fitting a
logistic dose response sigmoidal curve using OriginPro 8
(OriginLabs, Mass, USA).
TABLE-US-00006 TABLE 6 Inhibition of BACE-1 by Compounds of the
Invention Compound No. IC.sub.50 .mu.g/mL Heparin 0.002 (MW average
12 kDa) NAcLMWH 0.007 (MW average 4 kDa) 87 0.56 86 1.9 91 0.012 90
0.010 89 0.66 88 0.53 92 0.011 101 0.26 111 0.22 135 0.38 123 0.37
148 0.007 147 0.1 149 0.005
Example 5
Factor Xa Anticoagulant Assay Protocol
[0677] The compound of the invention, standard or control (5
.mu.l), is pipetted in assay buffer (0.9% sodium chloride) into a
96 well plate (Costar 3595) and 19 .mu.l of 0.03 IU/ml human
Antithrombin III (American Diagnostica Inc., product No. 433) in
assay buffer is added to each well. The plate is incubated for two
minutes at 37.degree. C. 19 .mu.l of bovine Factor Xa (14 nkat/ml;
Thermo Scientific; product No. 32521) in assay buffer is added to
each well and incubated for one minute at 37.degree. C. 19 .mu.l of
2.5 mM chromogenic substrate (American Diagnostica Inc. Spectrozyme
FXa Product No 222L) in assay buffer is added to each well and
incubated for 2 hours at 37.degree. C., followed by addition of 5
.mu.l of 30% acetic acid to each well. Absorbance at 405 nm is read
on a multiplate reader.
[0678] All compounds are tested in the dose range 0.004 to 50
.mu.g/ml and none display any measurable ability to accelerate
antithrombin-III mediated inactivation of Factor Xa, as measured by
cleavage of a peptide substrate.
[0679] Although the invention has been described by way of example,
it should be appreciated the variations or modifications may be
made without departing from the scope of the invention.
Furthermore, when known equivalents exist to specific features,
such equivalents are incorporated as if specifically referred to in
the specification.
INDUSTRIAL APPLICABILITY
[0680] The invention relates to compounds that are inhibitors of
BACE-1. The compounds are therefore indicated for the treatment or
prevention of diseases in which the inhibition of BACE-1 is
desirable, e.g. neurodegenerative disorders such as senile
dementia, pre-senile dementia, multi-infarct dementia or
Alzheimer's disease.
* * * * *