U.S. patent application number 12/926913 was filed with the patent office on 2011-08-18 for disaccharides for drug discovery.
This patent application is currently assigned to Alchemia Limited. Invention is credited to Giovani Abbenante, George Adamson, Bernd Becker, Nicholas B. Drinnan, Matthias Grathwohl, Giang Thanh Le, Wim Meutermans, Premraj Rajaratnam, Gerald Tometzki, Michael L. West.
Application Number | 20110201794 12/926913 |
Document ID | / |
Family ID | 3835711 |
Filed Date | 2011-08-18 |
United States Patent
Application |
20110201794 |
Kind Code |
A1 |
Meutermans; Wim ; et
al. |
August 18, 2011 |
Disaccharides for drug discovery
Abstract
Methods are described for the preparation of combinatorial
libraries of potentially biologically active disaccharide
compounds. These compounds are variously functionalized, with a
view to varying lipid solubility size, function an other
properties, with the particular aim of discovering novel drug or
drug-like compounds, or compounds with useful properties. The
invention provides intermediates, processes and synthetic
strategies for the solution or solid phase synthesis of
disaccharides, variously functionalized about the sugar ring,
including the addition of aromaticity and charge, and the placement
of pharmaceutically useful groups and isosteres.
Inventors: |
Meutermans; Wim; (Toowong,
AU) ; West; Michael L.; (Hemmant, AU) ;
Adamson; George; (Hampshire, GB) ; Le; Giang
Thanh; (Mt. Gravatt, AU) ; Drinnan; Nicholas B.;
(Highgate Hill, AU) ; Abbenante; Giovani;
(Samsonvale, AU) ; Becker; Bernd; (New Farm,
AU) ; Grathwohl; Matthias; (Konstanz, DE) ;
Rajaratnam; Premraj; (Eight Mile Plains, AU) ;
Tometzki; Gerald; (Manly West, AU) |
Assignee: |
Alchemia Limited
Eight Mile Plains
AU
|
Family ID: |
3835711 |
Appl. No.: |
12/926913 |
Filed: |
December 17, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10513286 |
Sep 27, 2005 |
7875707 |
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PCT/AU03/04947 |
Apr 24, 2003 |
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12926913 |
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Current U.S.
Class: |
536/17.5 ;
536/17.2; 536/29.1; 536/53; 536/55 |
Current CPC
Class: |
C40B 50/14 20130101;
C07H 3/04 20130101; C40B 40/12 20130101; C07H 3/02 20130101 |
Class at
Publication: |
536/17.5 ;
536/53; 536/17.2; 536/29.1; 536/55 |
International
Class: |
C07H 15/26 20060101
C07H015/26; C07H 1/00 20060101 C07H001/00; C07H 5/06 20060101
C07H005/06; C07H 17/02 20060101 C07H017/02 |
Foreign Application Data
Date |
Code |
Application Number |
May 3, 2002 |
AU |
PS 2138 |
Claims
1. A disaccharide compound of formula I A-d-L-e-B formula I wherein
A and B are independently chosen from ##STR00149## T is O or
CH.sub.2; R6 and R7 are hydrogen, or together form a carbonyl
oxygen; R1 is hydrogen, --N(Z)Y, C(Z)Y, OZ or SZ wherein; when R1
is N(Z)Y Y is selected from the group consisting of hydrogen or the
following; ##STR00150## wherein; Z is selected from hydrogen or X1,
Q is selected from hydrogen or W, W is selected from the group
consisting of alkyl, alkenyl, alkynyl, heteroalkyl, aryl,
heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms, X1 is
selected from the group consisting of alkyl, alkenyl, alkynyl,
heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl,
arylalkyl or heteroarylalkyl of 1 to 20 atoms, where R1 is C(Z)Y;
Y, is absent or is selected from hydrogen, double bond oxygen
(.dbd.O) to form a carbonyl, or triple bond nitrogen to form a
nitrile, Z is absent or is selected from hydrogen or X2, wherein X2
is selected from the group consisting of alkyl, alkenyl, alkynyl,
heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy, heteroaryloxy,
aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl,
acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or
heteroarylalkyl of 1 to 20 atoms, where R1 is OZ or SZ, Z is
selected from hydrogen or X3, wherein X3 is selected from the group
consisting of alkyl, alkenyl, alkynyl, heteroalkyl, acyl, arylacyl,
heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1
to 20 atoms, The groups R2, R3, R4 and R5 are selected from the
group consisting of hydrogen, N.sub.3, OH, OX4, N(Z)Y, wherein
N(Z)Y is as defined above or Y is ##STR00151## where Q and W are as
defined above, and X4 is independently selected from the group
consisting of alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl,
aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl,
aminoheteroaryl, alkylcarbamoyl, arylcarbamoyl or
heteroarylcarbamoyl, acyl, arylacyl, heteroarylacyl, aryl,
heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms, d and e
represent the connection points for A and B and replace one of the
groups R1, R2, R3, R4, or R5 in each of the groups A and B and form
the connection point for the linker L, d and e form a covalent bond
or are selected from the group consisting of: ##STR00152## L is
absent, or is selected from the group consisting of alkyl,
cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heteroalkyl,
cycloheteroalkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of
1 to 12 atoms.
2. The compound of claim 1, wherein when R1 is N(Z)Y, W is
substituted with at least one moiety selected from the group
consisting of OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen,
CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, and hydroxamic acid.
3. The compound of claim 1, wherein when R1 is N(Z)Y, Z and Y are
combined to form a monocyclic or bicyclic ring structure of 4 to 10
atoms.
4. The compound of claim 1, wherein X1 is substituted with at least
one moiety selected from the group consisting of OH, NO, NO.sub.2,
NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2, CH.sub.2F,
nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid,
carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl,
heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl,
aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate,
sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, and
hydroxamic acid.
5. The compound of claim 1, wherein; when R1 is C(Z)Y; X2 is
substituted with at least one moiety selected from the group
consisting of OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen,
CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, hydroxamic acid,
heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl,
thioaryl and thioheteroaryl.
6. The compound of claim 1, wherein; when R1 is C(Z)Y; Z and Y form
a ring structure of 4 to 10 atoms.
7. The compound of claim 6, wherein the ring structure is
substituted by X1 groups.
8. The compound of claim 1, wherein; when R1 is OZ or SZ; X3 is
substituted with at least one moiety selected from the group
consisting of OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen,
CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminodialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, hydroxamic acid,
heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl,
thioaryl and thioheteroaryl.
9. The compound of claim 1, wherein; when R1 is OZ or SZ; X4 is
substituted with at least one moiety selected from the group
consisting of OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen,
CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, and hydroxamic acid.
10. The compound of claim 1, wherein Z and Y are combined to form a
ring structure of 4 to 10 atoms.
11. The compound of claim 10, wherein the ring structure is
substituted with X1 groups.
12. The compound of claim 1, wherein L is substituted with at least
one moiety selected from the group consisting of OH, NO, NO.sub.2,
NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2, CH.sub.2F,
nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid,
carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl,
heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl,
aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate,
sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate,
hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl,
aminoheteroaryl, thioalkyl, thioaryl and thioheteroaryl.
13. The compound of claim 1, wherein in group A, T is oxygen, group
A is a pyranose ring, and the linker, d-L-e, is a glycosidic
linkage formed between the anomeric position R1 of group A, and any
position R1 to R5 of group B, such that the d is (--O--), L is
absent, and e is a covalent bond.
14. The compound of claim 13, wherein the structure of formula 1 is
##STR00153##
15. The compound of claim 13, wherein the structure of formula 1 is
##STR00154##
16. The compound of claim 13, wherein the structure of formula 1 is
##STR00155##
17. The compound of claim 13, wherein the structure of formula 1 is
##STR00156##
18. The compound of claim 13, wherein the structure of formula 1 is
##STR00157##
19. The compound of claim 13, wherein the structure of formula 1 is
##STR00158##
20. The compound of claim 13, wherein the structure of formula 1 is
##STR00159##
21. The compound of claim 13, wherein the structure of formula 1 is
##STR00160##
22. The compound of claim 13, wherein the structure of formula 1 is
##STR00161##
23. The compound of claim 1, wherein in group A, T is oxygen, group
A is a pyranose ring, the linker, d-L-e, forms an amide linkage in
which R6 and R7 of A is a C.dbd.O, R5 is d which is a covalent
bond, L is absent, and any of R1, R2, R3, R4, R5 on B is e which is
##STR00162##
24. The compound of claim 23, wherein the structure of formula 1 is
##STR00163##
25. The compound of claim 23, wherein the structure of formula 1 is
##STR00164##
26. The compound of claim 23, wherein the structure of formula 1 is
##STR00165##
27. The compound of claim 23, wherein the structure of formula 1 is
##STR00166##
28. The compound of claim 23, wherein the structure of formula 1 is
##STR00167##
29. The compound of claim 23, wherein the structure of formula 1 is
##STR00168##
30. The compound of claim 23, wherein the structure of formula 1 is
##STR00169##
31. The compound of claim 23, wherein the structure of formula 1 is
##STR00170##
32. The compound of claim 23, wherein the structure of formula 1 is
##STR00171##
33. The compound of claim 1, wherein in group A, T is oxygen, both
groups A and B are pyranose rings, the linkage, d-L-e, is an ether
type linkage in which any of R1 to R5 in group A and group B is d
and e respectively and is ##STR00172## and L is present.
34. The compound of claim 33, wherein the structure of formula 1 is
##STR00173##
35. The compound of claim 33, wherein the structure of formula 1 is
##STR00174##
36. The compound of claim 33, wherein the structure of formula 1 is
##STR00175##
37. The compound of claim 33, wherein the structure of formula 1 is
##STR00176##
38. The compound of claim 33, wherein the structure of formula 1 is
##STR00177##
39. The compound of claim 33, wherein the structure of formula 1 is
##STR00178##
40. The compound of claim 33, wherein the structure of formula 1 is
##STR00179##
41. The compound of claim 33, wherein the structure of formula 1 is
##STR00180##
42. The compound of claim 33, wherein the structure of formula 1 is
##STR00181##
43. The compound of claim 33, wherein the structure of formula 1 is
##STR00182##
44. The compound of claim 33, wherein the structure of formula 1 is
##STR00183##
45. The compound of claim 33, wherein the structure of formula 1 is
##STR00184##
46. The compound of claim 33, wherein the structure of formula 1 is
##STR00185##
47. The compound of claim 33, wherein the structure of formula 1 is
##STR00186##
48. The compound of claim 33, wherein the structure of formula 1 is
##STR00187##
49. The compound of claim 33, wherein the structure of formula 1 is
##STR00188##
50. The compound of claim 33, wherein the structure of formula 1 is
##STR00189##
51. The compound of claim 33, wherein the structure of formula 1 is
##STR00190##
52. The compound of claim 33, wherein the structure of formula 1 is
##STR00191##
53. The compound of claim 33, wherein the structure of formula 1 is
##STR00192##
54. The compound of claim 33, wherein the structure of formula 1 is
##STR00193##
55. The compound of claim 33, wherein the structure of formula 1 is
##STR00194##
56. The compound of claim 33, wherein the structure of formula 1 is
##STR00195##
57. The compound of claim 33, wherein the structure of formula 1 is
##STR00196##
58. The compound of claim 33, wherein the structure of formula 1 is
##STR00197##
59. The compound of claim 1, wherein in group A, T is oxygen, the
linkage, d-L-e, is a linkage in which R1 in group A is d, is
selected from the group consisting of a covalent bond, ##STR00198##
L is present; and e is selected from the group consisting of
##STR00199## and the connection to the B ring is at any of
R1-R5.
60. The compound of claim 59, wherein the structure of formula 1 is
##STR00200##
61. The compound of claim 59, wherein the structure of formula 1 is
##STR00201##
62. The compound of claim 59, wherein the structure of formula 1 is
##STR00202##
63. The compound of claim 59, wherein the structure of formula 1 is
##STR00203##
64. The compound of claim 1, wherein in group A, T is oxygen, the
linkage, d-L-e, is a linkage in which R1 in group A is d, R1 in
group B is e, and both d and e are independently chosen from the
group consisting of: a covalent bond; ##STR00204## and L is
present.
65. The compound of claim 64, wherein the structure of formula 1 is
##STR00205##
66. The compound of claim 64, wherein the structure of formula 1 is
##STR00206##
67. The compound of claim 1, wherein in group A, T is oxygen, the
linker, d-L-e, is a linkage in which at least one R group R1 to R5
in group A is d and is selected from the group consisting of
##STR00207## and any one of R1 to R5 in group B is e and e is
##STR00208## L is present.
68. The compound of claim 67, wherein the structure of formula 1 is
##STR00209##
69. The compound of claim 67, wherein the structure of formula 1 is
##STR00210##
70. The compound of claim 67, wherein the structure of formula 1 is
##STR00211##
71. The compound of claim 67, wherein the structure of formula 1 is
##STR00212##
72. The compound of claim 1, wherein in group A, T is oxygen, the
linkage, d-L-e, is a linkage in which at least one R group R1 to R5
in groups A and B is d and e and d and e are independently selected
from the group consisting of ##STR00213## and L is present.
73. The compound of claim 72, wherein the structure of formula 1 is
##STR00214##
74. The compound of claim 72, wherein the structure of formula 1 is
##STR00215##
75. The compound of claim 72, wherein the structure of formula 1 is
##STR00216##
76. The compound of claim 72, wherein the structure of formula 1 is
##STR00217##
77. The compound of claim 72, wherein the structure of formula 1 is
##STR00218##
78. A method of synthesizing a disaccharide compound of claim 1
comprising reacting compound A and compound B in solution.
79. A method of combinatorial synthesis of compounds of claim 1
comprising the step of immobilizing a compound of group B onto a
support through at least one of the functionalized positions R1 to
R5.
80. The method of claim 79, wherein the support is selected from
the group consisting of derivatised polystyrene, tentagel, wang
resin, MBHA resin, aminomethylpolystyrene, rink amide resin
DOX-mpeg, and polyethylene glycol.
81. A method of synthesising a compound of formula I in solution,
comprising the step of reacting compound A with a linker group L to
form a compound A-d-L and further reacting the compound A-d-L with
compound B to form a compound of formula I.
82. A method of synthesising a compound of formula I in solution,
comprising the step of reacting compound B with a linker group L to
form a compound B-e-L and further reacting the compound B-e-L with
compound A to form a compound of formula I.
Description
FIELD OF THE INVENTION
[0001] This invention relates to methods for the preparation of
combinatorial libraries of potentially biologically active
disaccharide compounds. These compounds are variously
functionalized, with a view to varying lipid solubility, size,
function and other properties, with the particular aim of
discovering novel drug or drug-like compounds, or compounds with
useful properties. The invention provides intermediates, processes
and synthetic strategies for the solution or solid phase synthesis
of disaccharides, variously functionalised about the sugar ring,
including the addition of aromaticity and charge, and the placement
of pharmaceutically useful groups and isosteres.
BACKGROUND OF THE INVENTION
[0002] From a drug discovery perspective, carbohydrate pyranose and
furanose rings and their derivatives are well suited as templates.
Each sugar represents a three-dimensional scaffold to which a
variety of substituents can be attached, usually via a scaffold
hydroxyl group, although occasionally a scaffold carboxyl or amino
group may be present for substitution. By varying the substituents,
their relative position on the sugar scaffold, and the type of
sugar to which the substituents are coupled, numerous highly
diverse structures are obtainable. An important feature to note
with carbohydrates, is that molecular diversity is achieved not
only in the type of substituents, but also in the three dimensional
presentation. The different stereoisomers of saccharides that occur
naturally (examples include glucose, galactose, mannose etc), offer
the inherent structural advantage of providing alternative
presentation of substituents.
##STR00001##
[0003] Although there are a number of examples of monosaccharides
being used as scaffolds for drug discovery purposes.sup.i,ii,iii,
there are only a limited number of examples of disaccharides or
higher saccharides being used as templates for the presentation of
pharmaceutically useful functional groups.
[0004] Derivatised disaccharides and higher saccharides, represent
a new class of compounds for drug discovery that are able to
address a significant and different group of receptors from those
addressed by monosaccharide scaffolds. This group or receptors can
be broadly described as those receptors in which the critical
binding groups are distal to each other. In principle,
monosaccharide scaffolds can be used to address up to five binding
groups (more usually 3 binding groups would be chosen), the
connection points on the scaffold are each separated by between 1
and 5 angstroms in space. Disaccharide scaffolds on the other hand
can accommodate up to eight binding groups although more usually
3-4 binding groups would be chosen, the connection points for each
of these groups being separated by as much as 10 angstroms in
space. Obviously the appended functional groups may be separated by
even greater distances in 3-dimensional space. The replacement of
the glycosidic bond linking the two monosaccharide components with
a spacer group can further increase the separation between binding
groups of interest.
[0005] The ability to address more distally placed binding groups
is an important feature for a number of biological receptor
molecules including the G-protein coupled receptors, where at the
extra-cellular opening to many of these receptors, the width of the
binding channel is up to 14 angstroms. Additionally, disaccharide
scaffolds can be used as probes of interactions which involve large
surface areas for example the protein-protein interaction of the
CD4-GP120 system, an important interaction in the aetiology of the
human immunodeficiency virus.
[0006] Through the development of a range of selectively protected
and modified monosaccharide, cyclitols and tetrahydropyran building
blocks, we have developed a system that allows the chemical
synthesis of highly structurally and functionally diverse
derivatised disaccharide and disaccharide analogue structures, of
both natural and unnatural origin. The diversity accessible is
particularly augmented by the juxtaposition of both structural and
functional aspects of the molecules. In order to access a wide
range of diverse structures, stereo-center inversion chemistry is
required, so as to achieve non-naturally occurring and hard to get
sugars and sugar analogues in a facile manner. Other chemistries
are also required that provide unnatural deoxy or deoxy amino
derivative which impart greater structural stability to the
drug-like target molecules. With a suite of reagents to effect a
suitable range of chemistries on a solid support, allowing such
things as; wide functional diversity, highly conserved
intermediates, a limited number of common building block to be
required, and with suitable chemistry to allow access to unusual
carbohydrate stereo-representations and including access to deoxy
and deoxy amino analogues, a methodology is then established that
can create focused libraries for a known target, or alternatively
diversity libraries for unknown targets for random screening.
[0007] It will be clearly understood that, although a number of
prior art publications are referred to herein, this reference does
not constitute an admission that any of these documents forms part
of the common general knowledge in the art, in Australia or in any
other country.
[0008] Many of the traditional methods of carbohydrate synthesis
have proved to be unsuitable to a combinatorial approach,
particularly because modern high-throughput synthetic systems
require procedures to be readily automatable. The compounds and
processes described herein are particularly suited to the solid and
solution phase combinatorial synthesis of carbohydrate-based
libraries, and are amenable to automation. The methods of the
invention yield common intermediates that are suitably
functionalized to provide diversity in the structure of the
compounds so generated. Using the method described, it is possible
to introduce varied functionality in order to modulate both the
biological activity and pharmacological properties of the compounds
generated.
[0009] Thus the compounds and methods disclosed herein provide the
ability to produce random or focused combinatorial-type libraries
for the discovery of other novel drug or drug-like compounds, or
compounds with other useful properties in an industrially practical
manner.
SUMMARY OF THE INVENTION
[0010] In a first aspect, the invention provides disaccharide
compounds of formula I
A-d-L-e-B formula I
[0011] In which the groups A and B are independently chosen
from
##STR00002##
[0012] in which the ring may be of any configuration and the
anomeric center where present may be of either the .alpha. or
.beta. configuration;
Independently for each ring
[0013] T may be O or CH.sub.2;
[0014] R6 and R7 are hydrogen, or together form a carbonyl
oxygen;
[0015] R1 may be hydrogen, --N(Z)Y, C(Z)Y, OZ or SZ wherein;
When R1 is N(Z)Y
[0016] Y is selected from hydrogen, or the following;
##STR00003##
[0017] Z is selected from hydrogen or X1;
[0018] Q is selected from hydrogen or W;
[0019] The groups Z and Y may be combined to form a monocyclic or
bicyclic ring structure of 4 to 10 atoms. This ring structure may
be further substituted with X1 groups;
[0020] The groups W are independently selected from alkyl, alkenyl,
alkynyl, heteroalkyl, aryl, heteroaryl, arylalkyl or
heteroarylalkyl of 1 to 20 atoms which is optionally substituted,
branched and/or linear. Typical substituents include but are not
limited to OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen, CF.sub.3,
CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, hydroxamic acid;
[0021] The groups X1 are independently selected from alkyl,
alkenyl, alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl,
aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms
which is optionally substituted, branched and/or linear. Typical
substituents include but are not limited to OH, NO, NO.sub.2,
NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2, CH.sub.2F,
nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid,
carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl,
heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl,
aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate,
sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate,
hydroxamic acid;
Where R1 is C(Z)Y;
[0022] Y, where present, is selected from hydrogen, double bond
oxygen (.dbd.O) to form a carbonyl, or triple bond nitrogen to form
a nitrile.
[0023] Z may be optionally absent, or is selected from hydrogen or
X2
[0024] Wherein X2 is independently selected from alkyl, alkenyl,
alkynyl, heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy,
heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or
thioheteroaryl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl,
arylalkyl or heteroarylalkyl of 1 to 20 atoms which is optionally
substituted, branched and/or linear. Typical substituents include
but are not limited to OH, NO, NO.sub.2, NH.sub.2, N.sub.3,
halogen, CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy,
amidine, guanidiniums, carboxylic acid, carboxylic acid ester,
carboxylic acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl,
aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl,
substituted or unsubstituted imine, sulfate, sulfonamide,
phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid,
heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl,
thioaryl or thioheteroaryl, which may optionally be further
substituted;
[0025] The groups Z and Y may be combined to form a monocyclic or
bicyclic ring structure of 4 to 10 atoms. This ring structure may
be further substituted with X1 groups;
Where R1 is OZ or SZ,
[0026] Z is selected from hydrogen or X3,
[0027] Wherein X3 is independently selected from alkyl, alkenyl,
alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl,
heteroaryl, arylalkyl or heteroaryialkyl of 1 to 20 atoms which is
optionally substituted, branched and/or linear. Typical
substituents include but are not limited to OH, NO, NO.sub.2,
NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2, CH.sub.2F,
nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid,
carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl,
heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl,
aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate,
sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate,
hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl,
aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may
optionally be further substituted;
[0028] The groups R2, R3, R4 and R5 are independently selected from
the group consisting of hydrogen, N.sub.3, OH, OX4, N(Z)Y, wherein
N(Z)Y is as defined above or additionally Y is
##STR00004##
where Q and W are as defined above, and X4 is independently
selected from alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl,
aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl,
aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, acyl,
arylacyl, heteroarylacyl, aryl, heteroaryl, aryalkyl or
heteroarylalkyl of 1 to 20 atoms which is optionally substituted,
branched and/or linear. Typical substituents include but are not
limited to OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen, CF.sub.3,
CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine,
guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic
acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, hydroxamic acid;
[0029] The groups Z and Y may be combined to form a monocyclic or
bicyclic ring structure of 4 to 10 atoms. This ring structure may
be further substituted with X1 groups;
The groups A and B are linked together with a linking structure
d-L-e, in which the groups d and e represent the connection points
for A and B and replace one of the groups R1, R2, R3, R4, or R5 in
each of the groups A and B and form the connection point for the
linker L. The groups d and e are independently chosen from a
covalent bond or the following list:
##STR00005##
L may be absent, or is selected from alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heteroalkyl, cycloheteroalkyl, aryl,
heteroaryl, arylalkyl or heteroarylalkyl of 1 to 12 atoms which is
optionally substituted, branched and/or linear, saturated or
unsaturated. Typical substituents include but are not limited to
OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2,
CH.sub.2F, nitrile, alkoxy, aryloxy, amidine, guanidiniums,
carboxylic acid, carboxylic acid ester, carboxylic acid amide,
aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl,
aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or
unsubstituted imine, sulfate, sulfonamide, phosphate,
phosphoramide, hydrazide, hydroxamate, hydroxamic acid,
heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl,
thioaryl or thioheteroaryl, which may optionally be further
substituted;
[0030] It is understood that the rules of molecular stoichiometry
will be upheld by the default addition of hydrogens atoms as
required.
A preferred embodiment of the first aspect provides for compounds
of formula I in which [0031] in group A, T is oxygen, [0032] group
A is a pyranose ring, [0033] The linker, d-L-e, is a glycosidic
linkage formed between the anomeric position R1 of group A, and any
position R1 to R5 of group B, such that the d is (--O--), L is
absent, and e is a covalent bond. [0034] Importantly, The R groups
on each ring may be selected independently from each other. For
example, R2 on ring A may be different from R2 on ring B. Exemplary
structure of this embodiment include but are not limited to:
##STR00006## ##STR00007##
[0034] Another preferred embodiment of the first aspect provides
for compounds of formula I in which [0035] In group A, T is oxygen,
[0036] group A is a pyranose ring, [0037] The linker, d-L-e, forms
an amide linkage in which R6 and R7 of A is a C.dbd.O, R5 is d
which is a covalent bond, L is absent, and any of R1, R2, R3, R4,
R5 on B is e which is
[0037] ##STR00008## [0038] Importantly, The R groups on each ring
may be selected independently from each other. For example, R2 on
ring A may be different from R2 on ring B. Exemplary structure of
this embodiment include but are not limited to:
##STR00009## ##STR00010##
[0038] Another preferred embodiment of the first aspect provides
for compounds of formula I in which [0039] in group A, T is oxygen,
[0040] both groups A and B are pyranose rings, [0041] The linkage,
d-L-e, is an ether type linkage in which any of R1 to R5 in group A
and group B is d and e respectively and is
[0041] ##STR00011## [0042] and L must be present. [0043]
Importantly, The R groups on each ring may be selected
independently from each other. For example, R2 on ring A may be
different from R2 on ring B. Exemplary structure of this embodiment
include but are not limited to:
##STR00012## ##STR00013## ##STR00014## ##STR00015##
##STR00016##
[0043] Another preferred embodiment of the first aspect provides
for compounds of formula I in which
[0044] In group A, T is oxygen, [0045] The linkage, d-L-e, is a
linkage in which R1 in group A is d, which is chosen from: a
covalent bond;
##STR00017##
[0046] L must be present;
[0047] and e is
##STR00018## [0048] Importantly, The R groups on each ring may be
selected independently from each other. For example, R2 on ring A
may be different from R2 on ring B. Exemplary structure of this
embodiment include but are not limited to:
##STR00019##
[0048] Another preferred embodiment of the first aspect provides
for compounds of formula I in which
[0049] In group A, T is oxygen, [0050] The linkage, d-L-e, is a
linkage in which R1 in group A is d, R1 in group B is e, and both d
and e are independently chosen from: a covalent bond;
##STR00020##
[0051] L must be present; [0052] Importantly, The R groups on each
ring may be selected independently from each other. For example, R2
on ring A may be different from R2 on ring B. Exemplary structures
of this embodiment include but are not limited to:
##STR00021##
[0052] Another preferred embodiment of the first aspect provides
for compounds of formula I in which
[0053] In group A, T is oxygen,
[0054] The linker, d-L-e, is a linkage in which any R group R1 to
R5 in group A may be d and is selected from
##STR00022##
And any R group R1 to R5 in group B may be e and e is
##STR00023##
L must be present. [0055] Importantly, The R groups on each ring
may be selected independently from each other. For example, R2 on
ring A may be different from R2 on ring B. [0056] Exemplary
structures of this embodiment include but are not limited to the
list below.
##STR00024##
[0056] Another preferred embodiment of the first aspect provides
for compounds of formula I in which
[0057] In group A, T is oxygen,
[0058] The linkage, d-L-e, is a linkage in which any R group R1 to
R5 in groups A and B may be d and e respectively and d and e are
independently selected from
##STR00025##
L must be present. [0059] Importantly, The R groups on each ring
may be selected independently from each other. For example, R2 on
ring A may be different from R2 on ring B. [0060] Exemplary
structures of this embodiment include but are not limited to the
list below.
##STR00026##
[0060] In a second aspect, the invention provides for a method of
synthesis of compounds of formula I comprising the step of reacting
two appropriately substituted and protected monosaccharide
compounds A and B in solution, In a third aspect, the invention
provides for a method of combinatorial synthesis of compounds of
the formula I comprising the step of immobilizing a compound of
group B onto a support through any of the functionalized positions
R1 to R5. Said support may be soluble or insoluble. Non-limiting
examples of insoluble supports include derivatised polystyrene,
tentagel, wang resin, MBHA resin, aminomethylpolystyrene, rink
amide resin etc. Non-limiting examples of soluble supports include
DOX-mpeg, polyethylene glycol etc. Compounds of the invention are
useful in screening for biological activity.
[0061] For the purposes of this specification it will be clearly
understood that the word "comprising" means "including but not
limited to", and that the word "comprises" has a corresponding
meaning.
General Method 1: Urea Formation
[0062] To the diamine (1.05 mmol) dissolved in
N,N-dimethylformamide (11 mL), was added isocyanate (1.99 mmol),
and the solution stirred for 2.5 h. Toluene (20 mL) was added and
all solvent removed to leave an oil. This procedure was repeated
twice more. This residue was then triturated with ether and the
resulting solid filtered to afford the product.
General Method 2: Transesterification
[0063] To a solution of the sugar (16.2 mmol) in a 1:1 mixture of
methanol/dichloromethane (110 mL) was added sodium methoxide (6.5
mmol) and the whole stirred under nitrogen for 2 h. Amberlite IR
120 H.sup.+ was added until pH 5 was reached. The resin was
filtered off and washed several times with methanol and the
combined filtrates were then concentrated to dryness to leave a
residue. The residue was either triturated with ether or purified
by column chromatography to give the desired product.
General Method 3: Azide Reduction
[0064] To a solution of the azido compound (0.30 mmol) in 4:1
N,N,-dimethylformamide/methanol (5 mL) was added a solution of
ammonium chloride (1.50 mmol) in water (0.5 mL). Activated zinc
dust (8.98 mmol) was then added and the suspension stirred for 40
min. A second addition of ammonium chloride (0.50 mmol) in water
(0.25 mL) and zinc dust (1.5 mmol) was made and the suspension
stirred for a further 40 min. After this time chloroform (50 mL)
was added and the suspension filtered through celite and washed
with chloroform/N,N,-dimethylformamide (1:1). These combined
filtrates were then washed with brine, dried (MgSO.sub.4), and all
solvent removed in vacuo to typically leave solid.
General Method 4: HBTU Coupling
[0065] To a solution of the acid (0.05 mmol) and HBTU (0.05 mmol)
in dry N,N,-dimethylformamide (0.2 mL) was added
diisoproplyethylamine (0.03 mL, 0.17 mmol) and the whole stirred
for 10 min. A solution of the sugar amine (0.04 mmol) in dry
N,N,-dimethylformamide (0.3 mL) was then added and the whole
stirred for 16 h. Chloroform (15 mL) was then added and washed with
water, 10% citric acid, saturated sodium hydrogen carbonate, brine,
dried (MgSO.sub.4), and the solvents removed in vacuo to leave an
oil. The crude was typically carried through to the next step
without further purification.
General Method 5: Global Deprotection for Formation of the Final
Product-1.
[0066] The sugar (0.04 mmol) was dissolved in a solution (3 mL) of
93% dry dichloromethane, 5% triethylsilane, 2% trifluoroacetic acid
and the reaction stirred at room temperature for 2 hours. The
solvents were then removed in vacuo and the solution then
freeze-dried to leave a white solid. This solid was then purified
by prep HPLC.
General Method 6: Global Deprotection for Formation of the Final
Product-2.
[0067] The sugar (0.0016 mmol) was dissolved in a solution (0.1 mL)
of 83% dry dichloromethane, 15% p-thiocresol, 2% trifluoroacetic
acid and the reaction stirred at room temperature for 0.5 hours.
The solvents were then removed in vacuo to give the crude
product.
General Method 7: DIC Coupling
[0068] To the acid (52 .mu.mol) and HOBT (52 .mu.mol) in dry DCM (1
mL) and dry DMF (1 drop) was added DIC (52 .mu.mol). The solution
was stirred for 1 min then added to a solution of amine (35
.mu.mol) in dry DCM (1 mL). The reaction mixture was stirred at
room temperature for 1 h then diluted with DCM, washed with 10%
citric acid, saturated hydrogen carbonate, brine, dried over
MgSO.sub.4 and the solvents removed in vacuo.
General Method 8: Ester Hydrolysis
[0069] To crude product 4 in dioxane (0.6 mL) was added 1M aq. KOH
(0.6 mL). The reaction mixture was stirred at room temperature for
1 h then concentrated in vacuo. The residue was dissolved in
CH.sub.3CN (2 mL) and DCM (0.5 mL) and stirred with Amberlite for 1
h. The solution was filtered and concentrated in vacuo to yield a
residue which was subsequently purified by prep HPLC.
General Method 9: Fmoc Cleavage Followed by DIC Coupling
[0070] Fmoc protected amino compound (.about.50 .mu.mol) was
dissolved in acetonitrile (2.4 mL) and piperidine (60 .mu.L, 0.60
.mu.mol) was added. The mixture was stirred over night, the
solvents evaporated in vacuo and the residue azeotroped with
toluene to afford a residue. The residue was taken up in dry
dichloromethane (2 mL) and a solution of octanoic acid (12 .mu.L,
75 .mu.mol) and DIC (12 .mu.L, 75 .mu.mol) in dry dichloromethane
(2 mL) (stirred for 5 min at room temperature prior to addition)
was added. Stirring was continued for 1 h and the mixture was
diluted with dichloromethane (50 mL), washed with 10% citric acid,
satd. sodium bicarbonate solution, filtered over a pad of cotton,
and the solvents removed in vacuo to afford the product as a crude
mixture.
General Method 10: Amine Deprotection
[0071] The fully protected block [2 mmol] was suspended in butanol
(15 ml) and ethylene diamine (15 ml) and the mixture heated at
reflux for 20 h. The solvents were evaporated, the residue taken up
in chloroform, washed with dilute brine, dried (MgSO.sub.4) and
evaporated. The compound was loaded onto a pad of silica with
chloroform and eluted with 9% methanol in chloroform to yield the
pure diamine quantitatively.
General Method 11: Diamine Coupling
[0072] The diamine (17 mg, 0.023 mmol) was dissolved in dry
chloroform (0.5 ml), DIPEA (3 mg, 4 .mu.l, 1 equiv) added and the
solution cooled to -78.degree. C. A solution of FMOC-Cl (4.2 mg,
0.7 equiv) in chloroform (0.2 ml) was added dropwise and allowed to
warm to it slowly before stirring for 16 h. The mixture was
partitioned between chloroform and water, the organic layer washed
with NaHCO.sub.3, brine, dried (MgSO.sub.4) and evaporated to
dryness. This gave the monoprotected amine as the major
product.
General Method 12: Thiourea Formation
[0073] To a solution of the sugar (0.48 mmol) in
N,N-dimethylformamide (4.8 mL), was added
4-fluorophenylisothiocyanate (0.48 mmol), and the solution stirred
at room temperature for 4 h. Toluene (5 mL) was added and all
solvent removed to leave an oil.
General Method 13: CBz Formation
[0074] To a solution of the sugar (0.48 mmol) in chloroform (4.8
mL), was added diisopropylethylamine (0.52 mmol). After 5 minutes
benzyl chloroformate (0.52 mmol) was added and the solution stirred
at room temperature for 2 h. Chloroform (5 mL) was then added and
washed with water, 10% citric acid, saturated sodium hydrogen
carbonate, brine, dried (MgSO.sub.4), and the solvents removed in
vacuo to leave an oil.
General Method 14: Sulphonamide Formation
[0075] To a solution of the sugar amine (0.0267 mmol) and
diisopropyethylamine (0.014 ml, 0.08 mmol) in dry dichloromethane
(0.25 mL), was added p-toluenesulfonyl chloride (10.2 mg, 0.054
mmol) and the solution stirred for 72 hrs. The reaction mixture was
diluted with dry dichloromethane (2 mL), washed with water (2 ml),
dried (MgSO.sub.4), and the solvents removed in vacuo to afford the
product as a crude mixture.
General Method 15: Acylation with Acetic Anhydride
[0076] To a solution of the sugar amine (0.04 mmol) and
diisopropyethylamine (0.02 ml, 0.12 mmol) in dry dichloromethane
(0.4 mL) was added dropwise acetic anhydride (0.015 mL, 0.12 mmol)
and the solution stirred for 1 h. The solution was concentrated in
vacuo to yield the crude product.
General Method 16: Fmoc Cleavage
[0077] Fmoc protected amino compound (0.48 mmol) was dissolved in
chloroform (4 mL) and piperidine (1 ml). The mixture was stirred
for 1 hr, evaporated to dryness. The residue was taken up in
acetonitrile (4 ml) and the solid was filtered off and the filter
cake washed with acetonitrile (1 ml). The acetonitrile solutions
were combined and evaporated to dryness. The residue was purified
by column chromatography (dichloromethane:methanol 20:1) to afford
the desired product,
General Method 17: Hydrolysis of Trifluoroacetate Ester and
Purification of the Final Products,
[0078] The sugar (0.04 mmol) was dissolved in a solution of
methanol (2 mL) and concentrated aqueous ammonium hydroxide (2 ml);
the reaction stirred at room temperature for 2 hours. The solvents
were then removed in vacuo and the solution then freeze-dried to
leave a white solid. This solid was then purified by prep HPLC.
EXAMPLE 1
Preparation of a Compound with a Glycosidic Linkage as Described by
Compounds of Formula II
##STR00027##
[0079] EXAMPLE 2
Preparation of a Compound with an Amide Linkage as Described by
Compounds of Formula III
##STR00028##
[0080] EXAMPLE 3
Preparation of a Compound with an Ether Linkage as Described by
Compounds of Formula IV
##STR00029##
[0081] EXAMPLE 4
Preparation of a Compound with an Ether Linkage as Described by
Compounds of Formula IV
##STR00030##
[0082] EXAMPLE 5
Preparation of a Compound with an Ether Linkage as Described by
Compounds of Formula V
##STR00031##
[0083] EXAMPLE 6
Preparation of a Compound with a Linkage as Described by Compounds
of Formula V and VI
##STR00032##
[0084] EXAMPLE 7
Preparation of a Compound with a Linkage as Described by Compounds
of Formula VI
##STR00033##
[0085] EXAMPLE 8
Preparation of a Compound with a Linkage as Described by Compounds
of Formula VII
##STR00034##
[0086] EXAMPLE 9
Preparation of a Compound with a Linkage as Described by Compounds
of Formula VIII
##STR00035##
[0087] EXAMPLE 10
Synthesis of 1,5-Anhydrogalactitol Acceptor
##STR00036##
[0088] 10-a. Synthesis of
1,5-anhydro-4-azido-2,4-dideoxy-2-DTPM-6-O-(4-methoxybenzyl)-D-galactitol
(56)
[0089] Compound 56 was prepared according to the procedure
described in General Method 2, .delta..sub.H (400 MHz; CDCl.sub.3)
3.25 (3H, s), 3.26 (3H, s), 3.65 (5H, m), 3.80 (3H, s), 4.09 (3H,
m), 4.50 (2H, q, J 9.5 Hz and J 3.6 Hz), 6.89 (2H, d, J 8.8 Hz),
7.26 (2H, d, J 8.8 Hz), 8.21 (1H, d, J 13.6 Hz), and 10.15 (1H, br
t, J 11.4 Hz); LCMS [M+H].sup.+ 475.
10-b. Synthesis of
2-amino-1,5-anhydro-4-azido-2,4-dideoxy-6-O-(4-methoxybenzyl)-D-galactito-
l (57)
[0090] To a solution of the sugar 56 (16.0 mmol) in a 3:1 mixture
of dry methanol/N,N,-dimethylformamide (120 mL) was added hydrazine
monohydrate (86.3 mmol) and the resulting reaction mixture was
stirred for 3 h. The resulting precipitate was removed by
filtration and the filtrate concentrated in vacuo. The residue was
then redissolved in dichloromethane, washed with saturated sodium
chloride, dried (MgSO.sub.4) and all solvent removed under reduced
pressure to leave a solid 57. The solid was used directly in the
next step.
10-c. Synthesis of
1,5-anhydro-4-azido-2-(3-carboxybenzyl)-2,4-dideoxy-6-O-(4-methoxybenzyl)-
-D-galactitol (58)
[0091] To a solution of the amine 57 (16.2 mmol) in methanol (55
ml), was added phthalic anhydride (216.2 mmol), and the whole
stirred for 2 h. The mixture was then evaporated to dryness under
reduced pressure and the residue azeotroped with toluene to leave a
cream solid 58.
10-d. Synthesis of
3-O-acetyl-1,5-anhydro-4-azido-2,4-dideoxy-6-O-(4-methoxybenzyl)-2-phthal-
limido-D-galactitol (59)
[0092] The acid 4 (16.3 mol) was suspended in dry pyridine (19 ml),
cooled to 0.degree. C., and acetic anhydride (48.7 mmol) added. The
suspension was then stirred for 1 h at 0.degree. C. followed by 18
h at room temperature. The solvent was then removed in vacuo and
the resulting residue azeotroped with toluene, redissolved in
chloroform and washed with water, 10% citric acid, saturated sodium
hydrogen carbonate, brine, dried (MgSO.sub.4) and the solvent
removed in vacuo to leave a yellow solid 59.
10-e. Synthesis of
1,5-Anhydro-4-azido-2,4-dideoxy-6-O-(4-methoxybenzyl)-2-phthallimido-D-ga-
lactitol (60)
[0093] Compound 60 was prepared according to the procedure
described in General Method 2, yield (77% from 55), .delta..sub.H
(400 MHz; CDCl.sub.3) 3.58 (1H, dd, J 14.3 Hz and J 7.3 Hz),
3.66-3.74 (1H, m), 3.82 (3H, s), 3.82-3.93 (1H, m), 4.03 (1H, t, J
11.4 Hz), 4.10 (1H, d, J 3.7 Hz), 4.42-4.52 (1H, m), 4.52 (2H, s),
4.66 (1H, dd, J 3.7 Hz and J 10.7 Hz), 6.90 (2H, d, J 8.9 Hz), 7.28
(2H, d, J 8.9 Hz), 7.74 (2H, m) and 7.67 (2H, m);
[M+NH.sub.4].sup.+=456.
EXAMPLE 11
Synthesis of a Trichloroacetimidate Donor
##STR00037##
[0094] 11-a. Thiomethyl
2-azido-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-.beta.-D-glucopyranoside
(62)
[0095] The thioglycoside 61 (21.2 mmol) was added in portions to a
suspension of sodium hydride (84.8 mmol) in DMF (106 ml) at
0.degree. C. After 20 min, the mixture was allowed to return to
room temperature and stirred for 30 min prior to recooling to
0.degree. C. To the suspension was then added 4-methoxybenzyl
chloride (11.5 ml) over 20 mins. The reaction mixture was then
allowed to return to room temperature and stirred for 16 h. The
resulting solution was cooled to 0.degree. C. and quenched with
ammonium chloride solution. The reaction mixture was partitioned
between water and chloroform, and the organic layer subsequently
washed with brine, dried (MgSO.sub.4) and evaporated. Residue was
purified by column chromatography to give the desired product 62 as
a solid in quantitative yield, .delta..sub.H (400 MHz; CDCl.sub.3)
2.22 (3H, s), 3.32-3.48 (3H, m), 3.54-3.70 (3H, m), 3.80 (3H, s),
3.81 (3H, s), 3.82 (3H, s), 4.17 (1H, d, J 9.5 Hz), 4.46 (1H, d, J
9.3 Hz), 4.49 (1H, d, J 10.3 Hz), 4.55 (1H, d, J 11.6 Hz), 4.63
(1H, d, J 5.2 Hz), 4.82 (2H, s), 6.75-6.95 (6H, m), 7.09 (2H, d, J
8.9 Hz), 7.24 (2H, d, J 8.9 Hz) and 7.30 (2H, d, J 10.3 Hz); LCMS
[M+Na].sup.+=618.2.
11-b. Synthesis of
2-Azido-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-D-glucopyranose
(63)
[0096] To a solution of thioglycoside 62 (112.9 mmol) in acetone
(450 mL) at 0.degree. C., shielded from light, was added water
(277.8 mmol), N-iodosuccinimide (134.9 mmol), followed by TMSOTf
(11.2 mmol), and the solution stirred for 90 min. Chloroform (400
mL) was added and the chloroform layer separated and washed with
saturated sodium hydrogen carbonate solution, saturated
Na.sub.2S.sub.2O.sub.3 solution, brine, dried (MgSO.sub.4) and
concentrated in vacuo to leave a solid. The solid was triturated
with ether to give the desired product 63 as a cream coloured solid
(98%), .delta..sub.H (400 MHz; CDCl.sub.3) 3.42 (1H, dd, J 3.0 Hz
and J 10.0 Hz), 3.52-3.66 (3H, m), 3.78 (3H, s), 3.81 (3H, s), 3.82
(3H, s), 3.97 (1H, t, J 9.8 Hz), 4.03 (1H, dq, J 2 Hz, J 4.9 Hz and
J 9.8 Hz), 4.42 (2H, t, J 10.3 Hz), 4.53 (1H, d, J 11.8 Hz), 4.62
(1H, br d, J 4.9 Hz), 4.73 (1H, d, J 10.6 Hz), 4.82 (2H, s),
6.81-6.93 (6H, m), 7.05 (2H, d, J 8.8 Hz), 7.24 (2H, d, J 11.7 Hz)
and 7.30 (2H, d, J 8.8 Hz); LCMS [M+Na].sup.+=588.3.
11-c. Synthesis of
2-Amino-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-D-glucopyranose
(64)
[0097] To a solution of the azide 63 (110.2 mmol) in dry DMF (275
mL) was added dithiothreitol (220.4 mmol). The reaction mixture was
then degassed with a stream of nitrogen, cooled to 0 C, and
triethylamine (220.4 mmol) added. The solution was then allowed to
return to room temperature and subsequently stirred for 24 h. The
solution was then diluted with ethyl acetate and washed with water,
brine, dried (MgSO.sub.4), the solvents removed in vacuo, and
resulting residue treated with diethyl ether (.about.250 mL) to
give the desired product 64 as a white solid (69%), LCMS
[M+H].sup.+=540.25.
11-d. Synthesis of
2-Deoxy-3,4,6-tri-O-(4-methoxybenzyl)-2-trifluoroacetamido-D-glucopyranos-
e (65)
[0098] To a solution of the amine 64 (75.6 mmol) in dry chloroform
(400 ml) at 0.degree. C. was added diisoproplyethylamine (113.4
mmol) and trifluoroacetic acid (16.0 mL, 113.4 mmol). The resulting
reaction mixture was then stirred for 30 min after which time the
reaction was allowed to return to room temperature and stir for a
further 1 h. At this time the reaction was cooled to 0.degree. C.
and further diisoproplyethylamine (113.4 mmol) and trifluoroacetic
acid (113.4 mmol) added, the reaction was allowed to return to room
temperature and stirred for 3 h. The reaction mixture was then
poured into dilute sodium hydrogen carbonate and the mixture
stirred for 30 min. The solid was washed with water, transferred to
a flask and dried by co-evaporation with acetonitrile to give the
crude product. Ethyl acetate was then added to the residue and the
resulting suspension refluxed for 2 h. Petrol ether (.about.300 mL)
was then added to this, the mixture cooled to room temperature, and
the resulting solid filtered and dried under high vacuum to give
the desired product 65 as a solid (77%), .delta..sub.H (400 MHz;
CDCl.sub.3) 3.04-3.17 (1H, m), 3.54-3.60 (2H, m), 3.61-3.71 (2H,
m), 3.79 (3H, s), 3.80 (3H, s), 3.81 (3H, s), 4.01 [dt, J 3.0 Hz
and J 10.4 Hz] and 4.16 [dt, J 3.7 Hz and J 10.1 Hz] (1H), 4.43
(2H, t, J 10.4 Hz), 4.50-4.60 (2H, m), 4.66-4.78 (2H, m), 5.23 (1H,
d, J 4.0 Hz), 6.11 (1H, br d, J 2.9 Hz), 6.80-6.90 (6H, m), 7.06
(2H, d, J 11.1 Hz), 7.18 (2H, d, J 12.6 Hz) and 7.24 (2H, d, J 11.1
Hz); LCMS [M+Na].sup.+=658.2.
11-f. Formation of Imidate (65)
[0099] To a solution of the lactol 11 (8.7 mmol) in tetrahydrofuran
(540 mL) was added trichloroacetonitrile (172.9 mmol), followed by
potassium carbonate (103.9 mmol), and the suspension stirred for 8
days. After 8 days the suspension was filtered through celite,
washed with tetrahydrofuran and all the solvent removed in vacuo to
leave a brown oil. The oil was then purified by column
chromatography (eluent toluene/acetone; 20:1) to give the desired
product 12 as a brown semi-solid (60%).
EXAMPLE 12
Synthesis of Disaccharide and Functionalisation
##STR00038## ##STR00039##
[0100] 12-a. Glycosylation with a Trichloroacetimidate Donor to
Afford Disaccharide (67)
[0101] To a solution of the acceptor 60 (1.8 mmol) and the donor 66
(2.7 mmol) in dry 1,2-dichloroethane (84 mL) was added 3 A
acid-washed molecular pellets (4 g) and the resulting mixture
stirred for 20 min. To the mixture was then added TMSOTf (1.8 mL of
a 0.1M solution in dry 1,2-dichloroethane, 0.18 mmol) and the
reaction then stirred for 15 min. After this time triethylamine (6
mL) was added and the suspension filtered, washed with
dichloromethane and all solvent removed in vacuo to leave a yellow
solid. This resulting solid was triturated with ether and filtered
to give the disaccharide 67 as a cream solid (65%), .delta..sub.H
(400 MHz; CDCl.sub.3) 3.26-3.35 (1H, m), 3.42-3.66 (5H, m),
3.68-3.72 (1H, m), 3.74 (3H, s), 3.76 (3H, s), 3.76-3.82 (4H, m),
3.78 (6H, s), 3.87-3.96 (1H, m), 4.32-4.54 (6H, m), 4.62-4.79 (3H,
m, [4.66, dd, J 3.4 Hz and J 13.3 Hz]), 5.02 (1H, dd, J 3.8 Hz and
J 12.6 Hz), 5.04 (1H, d, J 10.7 Hz), 6.34 (1H, d, J 10.3 Hz),
6.76-6.92 (8H, m), 7.07 (4H, dd, J 1.6 Hz and J 9.0 Hz), 7.20-7.24
(4H, m), 7.69-7.74 (2H, m) and 7.77-7.85 (2H, m); LCMS
[M+H+Na].sup.+=1078.2.
12-b. Amine Deprotection to Afford the Diamine Derivative (68)
[0102] The protected disaccharide 67 (0.95 mmol) was suspended in
n-butanol/ethylenediamine (1:1, 14 mL) and heated at reflux for 16
h. The solvent was then removed in vacuo, and the residue taken up
in chloroform, washed with dilute brine, dried (MgSO.sub.4), and
solvent removed in vacuo to leave an oil. The oil was purified by
column chromatography (eluent 10:1, chloroform/methanol) to give
the desired product 68 as a gummy solid [72%, used directly in next
step[, .delta..sub.H (400 MHz; CDCl.sub.3) 2.91 (1H, dd, J 8.2 and
J 10.3 Hz), 2.96-3.08 (1H, m), 3.21-3.29 (1H, m), 3.35-3.50 (4H,
m), 3.52-3.68 (5H, m), 3.74-3.82 (12H, singlets), 3.91-3.98 (1H,
m), 4.16 {1H, (d, J 3.6 Hz) and 4.32 (d, J 7.8 Hz)}, 4.35-4.52 (7H,
m), 4.58-4.64 (1H, m), 4.71 {1H, (d, J 10.3 Hz) and 4.91 (d, J 10.9
Hz)}, 6.80-6.92 (8H, m), 7.11 (2H, d, J 8.8 Hz), 7.2-7.32 (6H, m);
LCMS [M+H.sup.+]=830.35.
12-c. Urea Formation (69)
[0103] Compound 69 was prepared according to the procedure
described in General Method 1, yield [30%, used directly in next
step], LCMS [M+H].sup.+=1204.53.
12-d. Azide Reduction (70)
[0104] Compound 70 was prepared according to the procedure
described in General Method 3 [99%, used directly in next step],
LCMS [M+H].sup.+=1178.65.
12-e. Lipid HBTU Coupling (71)
[0105] Compound 71 was prepared according to the procedure
described in General Method 4, LCMS [M+H].sup.+=1374.83.
12-f. Global Deprotection (72)
[0106] Compound 72 was prepared according to the procedure
described in General Method 5, [30%, yield over two steps from
compound 70] as a white solid; LCMS [M+H].sup.+=894.22.
EXAMPLE 13
Synthesis of an Aminoacid Linked Lipidic Side Arm
##STR00040##
[0107] 13-a. DIC Coupling of a Aminoacid Side Arm (73)
[0108] Compound 73 (using 0.034 mmol of 70) was prepared according
to the procedure described in General Method 7; HPLC Method A,
Rt=7.5 min, [M+H].sup.+=1475.
13-b. Deprotection of the Boc Protected Amine and 4-Methoxybenzyl
Groups (74)
[0109] Compound 74 was prepared according to the procedure
described in General Method 5; HPLC Method A, Rt=4.82 min,
[M+H].sup.+=895.
13-c. DIC Coupling of a Lipidic Side Arm (75)
[0110] Compound 75 was prepared according to the procedure
described in General Method 7; HPLC-Method A, Rt=6.18 mins,
[M+H].sup.+=1117; RT=7.16 min, [M+H.sup.+]=1147.
13-d. Ester Hydrolysis to Provide the Free Acid (76)
[0111] Compound 75 was prepared according to the procedure
described in General Method 8, yield 25.7% from 70; HPLC Method A,
Rt=4.75 min, [M+H].sup.+=939.
EXAMPLE 14
Synthesis of an Aminoacid Linked Lipidic Side Arm-2
##STR00041## ##STR00042##
[0112] 14-a. DIC Coupling of a Aminoacid Side Arm (77)
[0113] Compound 77 was prepared (using 0.034 mmol of 70) according
to the procedure described in General Method 7, HPLC Method A,
Rt=7.3 mins, [M+H].sup.+=1497.
14-b. Deprotection of the Boc Protected Amine and 4-Methoxybenzyl
Groups (78)
[0114] Crude 77 was stirred at room temperature in DCM (2 mL), TFA
(40 .mu.L) and TES (100 .mu.L) for 2 hrs. The reaction mixture was
concentrated in vacuo, HPLC Method A, Rt=4.5 mins,
[M+H].sup.+=917.
14-c. DIC Coupling of a Lipidic Side Arm (79)
[0115] Compound 79 was prepared according to the procedure
described in General Method 7; HPLC Method A, Rt=5.15 mins,
[M+H].sup.+=1014.
14-d. Ester Hydrolysis to Provide the Free Acid (80)
[0116] Compound 79 was prepared according to the procedure
described in General Method 8, yield (from 70) 24.5%; HPLC-Method
A, Rt=4.50 mins, [M+H].sup.+=925.
EXAMPLE 15
Synthesis of
2-deoxy-2-(3-trifluoromethyl)-ureido-.beta.-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-4-(1-decanesulphonamido)-2-(3-trifluoromethyl)-ur-
eido-D-galactitol 4 with a Sulphonamide Lipidic Side Arm
##STR00043##
[0117] 15-a. Formation of a Sulphonamide Linked Lipidic Side Arm
(82)
[0118] To a solution of compound 70 (0.034 mmol) and pyridine (36.4
equiv) in dry dichloromethane (1 ml) was added 81 (20 equiv) in
several portions. The resulting reaction mixture was stirred under
nitrogen atmosphere for 2 hr and saturated sodium bicarbonate
solution (20 ml) added. After 30 min stirring, the aqueous phase
was extracted with dichloromethane; the combined organic solutions
were dried over MgSO.sub.4 and evaporated in vacuo to dryness. The
residue was purified by preparative TLC (eluent: neat EtOAc) to
furnish the crude
2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-2-(3-trifluoromethyl)-ureido-.beta.-
-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-6-(4-methoxybenzyl)-4-(1-decanesulphonamido)-2-(3-
-trifluoromethyl)-ureido-D-galactitol 82 (HPLC Method A, Rt=7.71
mins, [M+H].sup.+=1382.4). The product was used for the next step
without further purification.
15-b. Global Deprotection to Provide the Final Product (83)
[0119] Compound 83 was prepared according to the procedure
described in General Method 5, (10.8% from 70) HPLC Method A,
Rt=11.85 mins, [M+H].sup.+=902.53)
EXAMPLE 16
Synthesis of
2-deoxy-2-(3-trifluoromethyl)-ureido-.beta.-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-4-(1-decylphosphonamido)-2-(3-trifluoromethyl)-ur-
eido-D-galactitol 7
##STR00044##
[0120] 16-a. Formation of a Phosphonamide Linked Lipidic Side Arm
(85)
[0121] To a solution of 1-decylphosphonic acid (1 mmol) and
dimethylformamide (1.7 .mu.l) in dry dichloromethane (4 ml) was
added drop-wise oxalylchloride (261.2.mu.l, 3 mmol). The resultant
solution was stirred at room temperature under nitrogen atmosphere
for 1 hr and evaporated to dryness in vacuo. The residue was dried
under high vacuum for 1 hr to afford compound 84 as brownish
liquid. To a 84 (1 mmol) in DCM (4 ml) was added triethylamine (695
.mu.l, 5 mmol) followed by compound 70 [0.034 mmol]. The resultant
reaction mixture was stirred under nitrogen atmosphere for 1 hr and
1N hydrochloride solution (20 ml) added; the aqueous phase was
extracted with dichloromethane (3.times.20 ml); the combined
organic solutions were dried over MgSO4 and evaporated in vacuo to
dryness. The product was used for the next step without further
purification. (HPLC Method A, Rt=7.67 min,
[M+H].sup.+=1382.82).
16-b. Global Deprotection to Provide the Final Product (66)
[0122] Compound 86 was prepared according to the procedure
described in General Method 5, yield 8% (from 70); HPLC Method A,
Rt=5.36 mins, [M+H].sup.+=902.5)
EXAMPLE 17
Synthesis of
2-deoxy-2-(3-trifluoromethyl)-ureido-.beta.-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-4-(N-octanoylglycylamido)-2-(3-trifluoromethyl)-u-
reido-D-galactitol
##STR00045## ##STR00046##
[0123] 17-a. DIC Coupling to Form an Amide Linked Lipidic Side Arm
(88)
[0124] Compound 88 was prepared (from 0.034 mmol of 70) according
to the procedure described in General Method 7. The crude 88 was
used for the next step without further purification; HPLC Method A,
Rt=7.17 min; [M+H].sup.+=1361.80,
17-b. Global Deprotection to Provide the Final Product (89)
[0125] Compound 89 was prepared according to the procedure
described in General Method 5, yield 24% (from 70); HPLC Method A,
Rt=4.86 mins; [M+H].sup.+=881.36.
EXAMPLE 18
Synthesis of
2-deoxy-2-(3-trifluoromethyl)-ureido-.beta.-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-4-(N-octanoyl-N'-yl-3,4-dioxo-cyclobutene-1,2-dia-
mine)-2-(3-trifluoromethyl)-ureido-D-galactitol 92
##STR00047## ##STR00048##
[0126] 18-a. Coupling of Diethyl Squarate to Form a Vinylogous
Amide (90)
[0127] Amine 70 (0.034 mmol) was dissolved in Ethanol and DIPEA (35
mmol) and diethylsquarate (70 .mu.mol) was added. After stirring at
room temperature a complete conversion of the starting material to
5 was observed; HPLC Method A, Rt=6.70 mins;
[M+H].sup.+=1302.47.
18-b. Coupling of an Alkylamine Side Chain to the Squarate (91)
[0128] Decylamine (100 .mu.mol) was added in the mixture (from
18-a) and heated to 50.degree. C. over night. The mixture was
diluted with ethyl acetate, washed twice with 10% citric acid,
satd. sodium bicarbonate solution, and the solvents removed in
vacuo. The crude 91 was used without further purification in the
next step; HPLC Method A, Rt=7.66 mins, [M+H].sup.+=1413.15.
18-c. Global Deprotection to Provide the Final Product (92)
[0129] Compound 86 was prepared according to the procedure
described in General Method 5 to give pure 92, yield 26.2% (from
70); HPLC Method A, Rt=5.61 ml; [M+H].sup.+=933.32.
EXAMPLE 19
Synthesis of
2-deoxy-2-(3-trifluoromethyl)-ureido-5-D-glucopyranosyl
1,5-anhydro-2,4-dideoxy-4-((N-hexanoyl)-4-amino-butyroyl)-2-(3-trifluorom-
ethyl)-ureido-D-galactitol 10
##STR00049## ##STR00050##
[0130] 19-a. DIC Coupling to Form an Amide Linked Lipidic Side Arm
(94)
[0131] Compound 94 was prepared (from 0.034 mmol of 70) according
to the procedure described in General Method 7; HPLC Method A,
Rt=6.85 min; [M+H].sup.+=1361.47. The crude product was directly
used for the next step.
19-b. Global Deprotection to Provide the Final Product (95)
[0132] Compound 95 was prepared according to the procedure
described in General Method 5, to give pure 10, yield 42.1% (from
70); HPLC Method A, Rt=4.60 min; [M+H].sup.+=881.39.
EXAMPLE 20
Synthesis of a Disaccharide with a Lipidic Side Chain and Acid
Function
##STR00051## ##STR00052##
[0134] NB. Reaction series was completed separately for both the R
and the S isomers.
20-a. DIC Coupling to Form an Amide Linked Lipidic Side Arm
(96)
[0135] Compound 96 was prepared according to the procedure
described in General Method 7, the crude product was directly used
for the next step; HPLC Method A, Rt=7.70 mins;
[M+H].sup.+=1585.45.
20-b and 20-c. Fmoc Deprotection Followed by Amide Formation
(98)
[0136] Compound 98 was prepared according to the procedure
described in General Method 9, the crude product was directly used
for the general deprotection, (HPLC Method B, Rt=18.51 mins,
[M+H].sup.+=1489.77).
20-d. Global Cleavage to Afford the Final Product
[0137] Compound 99 was prepared according to the procedure
described in General Method 5, yield (R isomer) 27.5% from 70.
yield (S isomer) 10.5% from 70; HPLC Method A, Rt=4.88 mins;
[M+H].sup.+=953.46
EXAMPLE 21
Synthesis of a Disaccharide with a PEG Side Chain
##STR00053##
[0138] 21-a. DIC Coupling to Form an Amide Linked PEG Side Arm
(100)
[0139] Compound 100 was prepared according to the procedure
described in General Method 7; HPLC Method A, Rt=6.81 mins;
[M+H].sup.+=1338.49. The crude product was directly used for the
next step.
21-b. Global Cleavage to Afford the Final Product (101)
[0140] Compound 101 was prepared according to the procedure
described in General Method 5 (101 purified by preparative
chromatography on a C18 column), yield 21.8% (from 70); HPLC Method
A, Rt=4.27 min; [M+H].sup.+=958.52.
EXAMPLE 22
Synthesis of a Range of Lipid Conjugates
##STR00054##
[0141] 22-a. Preparation of Amido Derivatives at C-4 (102a, 102b,
102c, 102e)
[0142] Compounds 102a, 102b, 102c, 102e were prepared according to
the procedure provided in General Method 4.
22-b. Reaction with Acetic Anhydride (102d)
[0143] Added dropwise to a solution of the sugar amine (0.04 mmol)
In dry dichloromethane (0.4 mL) was acetic anhydride (0.12 mmol)
and the solution stirred for 16 h. Chloroform (15 mL) was then
added and washed with water, 10% citric acid, saturated sodium
hydrogen carbonate, brine, dried (MgSO.sub.4) and the solvent
removed under reduced pressure to give the title compound 5
quantitatively as an oil, LCMS [M+H].sup.+=1220.5.
22-c. Global Cleavage to Afford the Final Product (103a-103e)
[0144] Compound 103a-e were prepared according to the procedure
described in General Method 5.
TABLE-US-00001 TABLE 1 Derivatives prepared in Example 22.
##STR00055## Comp. No. R1 R2 Molecular ion Yield 102a R1a R2a [M +
H].sup.+ = 1276.2 87% 102b R1b R2a [M + H].sup.+ = 1374.2 Quant.
102c R1c R2a [M + H].sup.+ = 1358.3 Quant. 102d R1d R2a [M +
H].sup.+ = 1220.5 Quant. 102e R1e R2a [M + H].sup.+ = 1346.5 Quant.
103a R1a R2b [M + H].sup.+ = 796.3 50% 103b R1b R2b [M + H].sup.+ =
894.3 49% 103c R1c R2b [M + H].sup.+ = 878.3 30% 103d R1d R2b [M +
H].sup.+ = 740.1 30% 103e R1e R2b [M + H].sup.+ = 866.2 41% Side
Arms for Table 1. ##STR00056## ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062##
EXAMPLE 23
Synthesis of a Further Disaccharidic Library
[0145] Part 1: Preparation of Compounds 111-01 to 111-30
##STR00063## ##STR00064##
[0146] Part 2: Preparation of Compound 113-01 to 113-03
##STR00065##
[0147] Part 3: Preparation of Compounds 114-01-114-03 for the
Synthesis of Alternate Urea Building Blocks For the Preparation of
Further Derivatives at R.sub.3 as Exemplified by Structures
117-01-117-15 (by employing 5 different lipidic side chains).
##STR00066##
TABLE-US-00002 TABLE 2 Experimental Results, Intermediates and
Products for Example 23 ##STR00067## Rt Yield Product SM*
M.sup..sctn. R1 R2 R3 R4 [M + X].sup.+ (min).sup..dagger-dbl. % 68
67 10 I I L K [M + H].sup.+ = 830.5 6.07 74.8 104 67 10 I I M K [M
+ H].sup.+ = 804.5 5.36 18 105 68 11 H I L K [M + H].sup.+ =
1052.52 8.0-12.0 32 106-01 105 15 H N L K [M + H].sup.+ = 1094.6
6.90 100 106-02 68 11 H H L K [M + H].sup.+ = 1274.7 7.66 19.5
106-03 105 4 H C L K [M + H].sup.+ = 1224.7 7.32 84 106-04 105 14 H
D L K [M + Na].sup.+ = 1228.6 7.45 100 106-05 105 1 H E L K [M +
Na].sup.+ = 1261.7 7.38 78 106-06 105 14 H F L K [M + H].sup.+ =
1205.7 7.35 98 106-07 105 13 H G L K [M + Na].sup.+ = 1208.8 7.43
82 107-01 106-03 3 H C M K Detected in 108-01 form 100 107-02
106-04 3 H D M K Detected in 108-02 form 96 107-03 106-05 3 H E M K
Detected in 108-03 form 100 107-04 106-06 3 H F M K Detected in
108-04 form 100 107-05 106-07 3 H G M K Detected in 108-05 form 100
108-01 107-01 15 H C B K [M + H].sup.+ = 1240.7 6.99 100 108-02
107-02 15 H D B K [M + H].sup.+ = 1222.7 7.07 90 108-03 107-03 15 H
E B K [M + H].sup.+ = 1255.5 6.99 100 108-04 107-04 15 H F B K [M +
H].sup.+ = 1221.3 7.42 86 108-05 107-05 15 H G B K [M + Na].sup.+ =
1224.8 7.05 95 108-06 107-01 4 H C A K [M + H].sup.+ = 1366.9 8.09
85 108-07 107-02 4 H D A K [M + H].sup.+ = 1348.9 8.01 67.7 108-08
107-03 4 H E A K [M + H].sup.+ = 1381.9 8.33 88 108-09 107-04 4 H F
A K [M + H].sup.+ = 1347.4 7.86 100 108-10 107-05 4 H G A K [M +
H].sup.+ = 1328.6 8.18 100 109-01 108-06 16 I C A K Detected in
110-02 form 67 109-02 108-07 16 I D A K Detected in 110-03 form 61
109-03 108-08 16 I E A K Detected in 110-04 form 90 109-04 108-09
16 I F A K Detected in 110-05 form 58 109-05 108-10 16 I G A K
Detected in 110-06 form 51 110-01 68 15 N N L K [M + H].sup.+ =
914.5 5.71 100 110-02 109-01 15 N C A K [M + H].sup.+ = 1186.76
7.50 100 110-03 109-02 15 N D A K [M + H].sup.+ = 1168.77 7.54 100
110-04 109-03 15 N E A K [M + H].sup.+ = 1201.8 7.33 100 110-05
109-04 15 N F A K [M + H].sup.+ = 1167.5 7.30 100 110-06 109-05 15
N G A K [M + H].sup.+ = 1148.5 7.51 100 110-07 109-01 4 C C A K [M
+ H].sup.+ = 1316.7 7.59 86.8 110-08 109-02 4 C D A K [M + H].sup.+
= 1298.6 7.94 100 110-09 109-03 4 C E A K [M + H].sup.+ = 1331.71
7.90 100 110-10 109-04 4 C F A K [M + H].sup.+ = 1297.5 7.58 100
110-11 109-05 4 C G A K [M + H].sup.+ = 1278.7 7.82 100 110-12
109-01 14 D C A K [M + H].sup.+ = 1298.5 7.68 84.7 110-13 109-02 14
D D A K [M + H].sup.+ = 1280.4 7.92 100 110-14 109-03 14 D E A K [M
+ H].sup.+ = 1313.83 8.03 100 110-15 109-05 14 D G A K [M +
H].sup.+ = 1260.5 7.89 28.2 110-16 109-01 1 E C A K [M + Na].sup.+
= 1353.8 7.86 100 110-17 109-02 1 E D A K [M + H].sup.+ = 1313.81
7.81 100 110-18 109-04 1 E F A K [M + H].sup.+ = 1312.8 7.64 100
110-19 109-05 1 E G A K [M + H].sup.+ = 1293.8 7.88 100 110-20
109-01 12 F C A K [M + H].sup.+ = 1297.9 7.60 80 110-21 109-02 12 F
D A K [M + H].sup.+ = 1279.8 7.92 77 110-22 109-03 12 F E A K [M +
H].sup.+ = 1312.84 7.65 100 110-23 109-04 12 F F A K [M + H].sup.+
= 1278.8 7.73 73 110-24 109-05 12 F G A K [M + H].sup.+ = 1259.9
7.64 70 110-25 109-01 13 G C A K [M + H].sup.+ = 1278.8 7.64 66
110-26 109-02 13 G D A K [M + H].sup.+ = 1260.8 7.91 75 110-27
109-03 13 G E A K [M + H].sup.+ = 1293.87 7.96 100 110-28 109-04 13
G F A K [M + H].sup.+ = 1259.8 7.75 72 110-29 109-05 13 G G A K [M
+ H].sup.+ = 1240.8 7.67 80 110-30 109-03 1 O E A K Compound
fragments to 110-31 100 under LCMS conditions 110-31 P E A K [M +
H].sup.+ = 1202.81 7.26 111-01 110-07 5 C C A I [M + H].sup.+ =
836.4 5.33 100 111-02 110-08 5 C D A I [M + H].sup.+ = 818.4 5.57
100 111-03 110-09 5 C E A I [M + H].sup.+ = 851.5 5.43 100 111-04
110-10 6 C F A I [M + H].sup.+ = 817.4 5.44 18 111-05 110-11 5 C G
A I [M + H].sup.+ = 798.5 5.54 100 111-06 110-12 5 D C A I [M +
H].sup.+ = 818.4 5.56 100 111-07 110-13 4 D D A I [M + H].sup.+ =
800.4 5.47 100 111-08 110-14 5 D E A I [M + H].sup.+ = 833.4 5.83
100 111-09 110-15 5 D G A I [M + H].sup.+ = 780.4 5.41 100 111-10
110-16 5 E C A I [M + H].sup.+ = 851.3 5.64 100 111-11 110-17 5 E D
A I [M + H].sup.+ = 833.3 5.75 100 111-12 110-18 5 E F A I [M +
H].sup.+ = 832.5 5.53 16 111-13 110-19 5 E G A I [M + H].sup.+ =
813.3 5.69 100 111-14 110-20 6 F C A I [M + H].sup.+ = 817.4 5.49
51 111-15 110-21 6 F D A I [M + H].sup.+ = 799.4 5.46 48 111-16
110-22 5 F E A I [M + H].sup.+ = 832.5 5.65 2 111-17 110-23 6 F F A
I [M + H].sup.+ = 798.4 5.36 20 111-18 110-24 6 F G A I [M +
H].sup.+ = 779.5 5.58 53 111-19 110-25 5 G C A I [M + H].sup.+ =
798.5 5.30 100 111-20 110-26 5 G D A I [M + H].sup.+ = 780.5 5.57
100 111-21 110-27 5 G E A I [M + H].sup.+ = 813.4 5.69 100 111-22
110-28 6 G F A I [M + H].sup.+ = 779.5 5.08 10 111-23 110-29 5 G G
A I [M + H].sup.+ = 760.5 5.28 100 111-24 108-06 5 H C A I [M +
H].sup.+ = 886.55 6.07 100 111-25 108-07 5 H D A I [M + H].sup.+ =
868.58 5.99 100 111-26 108-08 5 H E A I [M + H].sup.+ = 901.4 6.00
100 111-27 108-09 6 H F A I [M + H].sup.+ = 867.6 6.03 20 111-28
108-10 5 H G A I [M + H].sup.+ = 848.6 5.91 100 111-29 110-30 5 P E
A I [M + H].sup.+ = 722.4 4.63 100 111-30 109-03 5 I E A I [M +
H].sup.+ = 679.5 4.62 100 112-01 104 15 N N B K [M + Na].sup.+ =
952.7 5.01 47 112-02 104 4 C C Q K [M + H].sup.+ = 1320.5 6.99 76
112-03 104 1 E E R K [M + H].sup.+ = 1365.4 7.21 92 113-01 112-01 5
N N B I [M + H].sup.+ = 450.26 0.69 100 113-02 112-02 5 C C Q I [M
+ H].sup.+ = 840.4 4.53 100 113-03 104 5 I I M I [M + H].sup.+ =
324.2 0.62 100 114-01 68 1 S S L K [M + H].sup.+ = 1204.9 7.02 100
114-02 68 1 T T L K [M + H].sup.+ = 1096.5 6.82 100 115-03 68 1 U U
L K [M + H].sup.+ = 1204.3 7.25 100 111-01-1 111-01 17 C C A I As
Start Material N/A 43 111-02-1 111-02 17 C D A I As Start Material
N/A 19.2 111-06-1 111-06 17 D C A I As Start Material N/A 34.5
111-07-1 111-07 17 D D A I As Start Material N/A 51 111-08-1 111-08
17 D E A I As Start Material N/A 27 111-10-1 111-10 17 E C A I As
Start Material N/A 16 111-11-1 111-11 17 E D A I As Start Material
N/A 75 111-29-1 111-29 17 P E A I As Start Material N/A 45 SM* =
Starting Material M.sup..sctn. = Method of Synthesis (General
Method) Rt (min).sup..dagger-dbl.: All compounds in Table 2 were
analysed by HPLC Method A. Note: Under the employed analytical
conditions, compounds containing amino group (compounds classed as
68, 104, 105, 107, 109) elute in unusual broad peaks, sometimes
several minutes wide; therefore, most of the time they are detected
as acetamide derivatives obtained via acylating with acetic
anhydride.] Substituents for Table 2 ##STR00068## ##STR00069##
##STR00070## ##STR00071## ##STR00072## ##STR00073## ##STR00074##
##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079##
##STR00080## ##STR00081## ##STR00082## ##STR00083## ##STR00084##
##STR00085## ##STR00086## ##STR00087## ##STR00088##
##STR00089##
HPLC Methods.
HPLC Method A
TABLE-US-00003 [0148] Flow Rate Time H.sub.2O % MeCN % mL/min 0 95
5 2 1 95 5 2 7 0 100 2 12 0 100 2
[0149] Agilent SB Zorbax C18 4.6.times.50 mm (5 .mu.m, 80
.ANG.)
[0150] LC Mobile Phase: Acetonitrile:Water 0.1% formic acid
HPLC Method B
TABLE-US-00004 [0151] Flow Rate Time H.sub.2O % MeCN % mL/min 0.00
95 5 1.00 95 5 20.00 0 100
[0152] Agilent SB Zorbax C18 4.6.times.50 mm (5 .mu.m, 80
.ANG.)
[0153] LC Mobile Phase: Acetonitrile:Water 0.1% formic acid
EXAMPLE 24
Synthesis of an Alpha 1.fwdarw.4 Linked Disaccharidic Compound
Selective Removal of Protecting Groups for Diversity-Part 1
##STR00090##
[0154] 24-a: Glycosylation
[0155] Compound 118 [1.0 mmol] and 60 [1.5 mmol] were dissolved in
dry dichloromethane [16 mL] and stirred with molecular sieves [4
.ANG. acid washed] at room temperature for 1 h. To the mixture was
then added 2,6-di-tert.-butylpyridine [1.6 mmol] and DMTST [1.6
mmol], and the reaction stirred at room temperature. After 2.5 h
further 60 [0.2 mmol], 2,6-di-tert.-butylpyridine [0.2 mmol] and
DMTST [0.2 mmol] were added and the reaction stirred at room
temperature for 1 h. The reaction was then quenched with
triethylamine, the solvents were removed in vacuo and the product
purified by column chromatography (silica, petrolether/ethyl
acetate 2:1) to give 119 as a colourless foam [63%]; HPLC Method A,
Rt=8.09 mins, [M+Na].sup.+=1087.56; 1H-NMR (CDCl.sub.3): 8.00 (d,
2H, Ar), 7.82-7.65 (m, 2H, Ar), 7.95-7.14 (m, 19H, Ar), 6.90 (d,
2H, Ar), 5.52 (dd, 1H, H-2', J1'-2'=3.4 Hz, J2'-3'=10.1 Hz), 5.24
(d, 1H, H1'), 4.82 (s, 1H, CHPh), 4.72-4.63 (m, 2H), 4.54 (dd, 1H,
H-3', J3'-4'=3.6 Hz), 4.38 (AB, 2H, CH.sub.2Ar, Jgem=11.7 Hz),
3.95-3.86 (m, 2H), 3.83 (s, 3H, OCH.sub.3), 3.83-3.76 (m, 1H),
3.62-3.56 (m, 1H), 3.49 (dd, 1H, J=5.9 Hz, J=9.0 Hz), 3.38 (dd, 1H,
J=7.7 Hz, J=9.2 Hz), 3.37 (dd, 1H, H4', J4'-5'<1 Hz), 3.30 (dd,
1H, H1-a, Jgem=12.5 Hz. J1a,2=1.9 Hz), 3.25 (m, 1H), 2.92 (dd, 1H,
H-1b, J1b,2<1 ), 0.88 (s, 9H, tBu),
24-b: Azide Reduction
[0156] Compound 119 [0.164 mmol] was treated according to the
procedure described in General Method 3 (NB: compound smears over
several minutes on HPLC column (HPLC Method A);
[M+H].sup.+=1039.53. The solution of the crude product 120 was
directly used for the next conversion.
24-c: Amide Coupling
[0157] Crude 120 was treated with undecanoic acid 120 mg [0.65
mmol] according to the procedure described by General Method 7. 1
The residue containing 121 was purified on a silica column
(gradient petrolether/ethyl acetate 2:1 to petrolether/ethyl
acetate 1:1 with 2% triethylamine) to give 121 [51%]; HPLC Method
A, Rt=9.14 mins; [M+Na].sup.+=1229.69, and unreacted 120 (31%).
24-d: Silylether Cleavage
[0158] To a solution of 121 in DMF [3 mL], was added a 1 molar
solution of TBAF in THF [0.5 mL] and acetic acid [30 .mu.L], and
the mixture heated to 65.degree. C. for 6 h. The reaction mixture
was diluted with ethyl acetate and the solution washed with
saturated sodium bicarbonate solution and water, the dried over
magnesium sulfate and the solvents removed in vacuo to give crude
122 (100% conversion by ELSD); HPLC Method A, Rt=7.17 mins;
[M+H].sup.+=969.55.
24-e to e2: Removal of Acid Labile Protecting Groups
[0159] (24-e1) Crude 122 was dissolved in dry dichloromethane [5
mL] and triethylsilane [0.5 mL] and trifluoroacetic acid [0.1 mL]
were added. After stirring at room temperature for 10 min
conversion to 123a was complete (HPLC Method A, Rt=6.80 min,
[M+H].sup.+=849.50). (24-e2). Further stirring at room temperature
for 3 h gave 123 (100% conversion by ELSD); HPLC Method A, Rt=6.71
mins; [M+H].sup.+=761.43.
Selective Removal of Protecting Groups for Diversity-Part 2
##STR00091##
[0160] 24-f1 to f2: Removal of Acid Labile Protecting Groups
[0161] (24-f1). Compound 121 [8 .mu.mol] was dissolved in dry
dichloromethane [1 mL] and triethylsilane [0.1 mL] and
trifluoroacetic acid [0.02 mL] were added. After stirring at room
temperature for 2 min conversion to 124a was complete (HPLC Method
A, Rt=8.54 mins, [M+H].sup.+=1187.49). (24-f2). Further stirring at
room temperature for 3 hrs gave 124 (100% conversion by ELSD); HPLC
Method A, Rt=7.87 mins, [M+H].sup.+=999.56.
Selective Removal of Protecting Groups for Diversity-Part 3
##STR00092##
[0163] 24-g: Phthalimido Cleavage
[0164] To a solution of 121 [0.10 mmol] in ethanol [5 mL] was added
hydrazine hydrate [0.05 mL] and the solution refluxed for 20 h. The
solvents were removed in vacuo and the residue co-evaporated with
toluene to give crude 125 (100% conversion by ELSD). Product smears
over several minutes on HPLC (HPLC Method A),
[M+H].sup.+=1077.43.
24-g: Sulfonamide Formation
[0165] Compound 125 [1.8 .mu.mol] was reacted with tosylchloride [5
mg] according to General Method 14 to give 126 (100% conversion by
ELSD), HPLC Method A, Rt=9.25 mins; [M+H].sup.+=1231.65,
[M+Na].sup.+=1253.63.
Selective Removal of Protecting Groups for Diversity-Part 4
##STR00093## ##STR00094##
[0166] 24-i: Urea Formation
[0167] Compound 125 [0.10 mmol] was reacted with
3-trifluoromethylphenyl isocyanate [0.4 mmol] according to General
Method 1 to afford 127 (73% purity by ELSD); HPLC Method A, Rt=9.04
mins; [M+H].sup.+=1264.75.
24-j: Ester Cleavage
[0168] Compound 127 [0.10 mmol] was treated according to the
procedure described in General Method 2 (with the exception that
only MeOH was used as solvent) to afford crude 128; HPLC Method A,
Rt=8.60 mins; [M+H].sup.+=1126.48, [M+H].sup.+=1148.46.
24-k: Carbamate Formation
[0169] To a solution of 128 [0.10 mmol] in dry DMF [5 mL], was
added 3-trifluoromethylphenyl isocyanate [0.4 mmol] and DBU [45
.mu.L], and the solution heated to 80.degree. C. for 20 h. The
reaction mixture was diluted with ethyl acetate, washed with
saturated sodium bicarbonate solution and water, dried over
magnesium sulfate, and the solvents removed in vacuo to give
129.
24-l: Silylether Cleavage
[0170] To a solution of 129 in DMF [3 mL], was added a 1M solution
of TBAF in THF [0.5 mL] and acetic acid [30 .mu.L], and the
reaction mixture heated to 65.degree. C. for 6 h. The reaction
mixture was diluted with ethyl acetate and the solution washed with
saturated sodium bicarbonate solution, water, dried over magnesium
sulfate, and the solvents removed in vacuo to give crude 130.
24-m1 to 24-m2: Removal of Acid Labile Protecting Groups
[0171] (24-m1). To a solution of compound 130 in dichloromethane [5
mL] was added triethylsilane [0.5 mL] and trifluoroacetic acid [0.1
mL]. After stirring at room temperature for 2 min conversion to
131a was complete, (24-m2). Further stirring at room temperature
for 3 hrs gave 131 quantitatively.
Selective Removal of Protecting Groups for Diversity-Part 5
##STR00095##
[0172] 24-n: Silylether Cleavage
[0173] To a solution of 127 in DMF [3 mL], was added a 1M solution
of TBAF in THF [0.5 mL] and acetic acid [30 .mu.L], and the mixture
heated to 65.degree. C. for 6 h. The reaction mixture was then
diluted with ethylacetate and the solution washed with saturated
sodium bicarbonate solution, water, dried over magnesium sulfate,
and the solvents removed in vacuo to give crude 132.
24-o: Carbamate Formation
[0174] To a solution of 132 [0.10 mmol] in DMF [5 mL] was added
3-trifluoromethylphenyl isocyanate [0.4 mmol] and DBU [45 .mu.L],
and the solution heated to 80.degree. C. for 20 h. The reaction
mixture was diluted with ethyl acetate, washed with saturated
sodium bicarbonate solution, water, dried over magnesium sulfate,
and the solvents removed in vacuo to give 133.
24-p: Ester Cleavage
[0175] Compound 133 [0.10 mmol] was treated according to General
Method 2 (with the exception that only MeOH was used as solvent) to
provide crude 134.
24-q1 to 24-q2: Removal of Acid Labile Protecting Groups
[0176] (24-q1). Compound 134 was dissolved in dry dichloromethane
[5 mL] and triethylsilane [0.5 mL] and trifluoroacetic acid [0.1
mL] were added. After stirring at room temperature for 2 min
conversion to 135a was complete. (24-q2). Further stirring at room
temperature for 3 hrs gave 135.
EXAMPLE 25
Synthesis of a Beta 1.fwdarw.6 Linked Disaccharidic Compound for
Drug Discovery-1
##STR00096##
[0177] 25-a: Glycosylation
[0178] To a solution of 60 [0.48 mmol] and 136 [0.70 mmol] in
dichloroethane [8 mL] was added DMTST [0.48 mmol], and the mixture
stirred for 45 mins at room temperature. At that time further DMTST
[0.21 mmol] was added, and after stirring for 20 mins the reaction
was quenched by the addition of triethylamine [0.5 mL]. The
reaction mixture was then diluted with dichloromethane, washed with
10% citric acid, saturated sodium bicarbonate solution, dried over
magnesium sulfate, the solvents evaporated in vacuo and the product
purified by column chromatography (silica, petrol ether/ethyl
acetate 1:1) to give 137 as a colorless foam [59%]; HPLC Method A,
Rt=5.06 mins: [M+Na].sup.+=758.49.
25-b: Acetylation
[0179] To a solution of 137 [2 mg] in pyridine [0.2 ml] was added
acetic anhydride [50 .mu.L], and the ensuing reaction mixture
stirred at room temperature for 2 hrs. The reaction mixture was
then diluted with dichloromethane and washed with 10% citric acid,
saturated sodium bicarbonate solution, dried over magnesium sulfate
and the solvents evaporated in vacuo to give 138; .sup.1H-NMR
(CDCl.sub.3): 7.85-7.62 (m, 8H, Ar), 5.75 (dd, 1H, H-3',
J.sub.2',3'=10.4 Hz, J.sub.3',4'=8.8 Hz), 5.70 (dd, 1H, H-3,
J.sub.2,3=11.0 Hz, J.sub.3,4=3.4 Hz), 5.39 (d, 1H, H-1',
J.sub.1',2'=8.7 Hz), 5.12 (dd, 1H, H-4', J.sub.4',5'<1 Hz), 4.64
(ddd, 1H, H-2), 4.25 (dd, 1H, H-2'), 4.00 (dd, 1H, H-4), 3.88-3.76
(m, 3H, H-1a, H-5, H-5'), 3.70-3.52 (m, 5H, H-1b, H-6a, H-6b,
H-6a', H-6b'), 2.07, 1.98, 1.87, 1.71 (each s, 3H, Ac).
EXAMPLE 26
Synthesis of a Beta 1.fwdarw.46 Linked Disaccharidic Compound for
Drug Discovery-2
##STR00097##
[0180] 26-a. Formation of a Beta 1.fwdarw.6 Linked Disaccharide
[0181] A solution of thioglycoside 139 [0.154 mmol],
trichloroacetimidate 66 [1.5 eq. with respect to thioglycoside] and
4 angstrom molecular sieves [0.26 g] in 1,2-DCE [2.6 mL] was
stirred at room temperature for 15 mins. At this time TMSOTf [0.3
eq.] was added. The reaction was allowed to stir for 30 mins at
which time it was quenched by the addition of triethylamine [2 mL].
The reaction mixture was diluted with DCM, filtered and the
resulting filtrate concentrated in vacuo to afford a residue. The
residue was purified by column chromatography [toluene/acetone,
20:1] to provide the product as a colourless foam [58%]; HPLC
Method A, Rt=8.02 mins; [M+Na].sup.+=1316; .sup.1H-NMR, CDCl.sub.3
.delta. 4.07 (d, 1-H, H-1a J.sub.1,2=9.2 Hz), 4.42 (d, 1-H, H-1b,
J.sub.1,2=8.2 Hz) indicating two beta linkages.
EXAMPLE 27
Synthesis of a Methyl Glycoside Disaccharide for Drug Discovery
##STR00098##
[0182] 27-a. Benzoylation of the 3-OH Position
[0183] To a solution of 141 [37.2 mmol] in 1,2-dichloromethane [140
mL] at 0.degree. C. was added DMAP [2 eq.] and benzoyl chloride
[1.5 eq.]. The reaction mixture was allowed to return to room
temperature, and stirred for 2 hrs. Methanol was added and the
reaction mixture was stirred for a further 15 mins. The reaction
mixture was then diluted with CHCl.sub.3, washed with 10% citric
acid solution, saturated NaHCO.sub.3 solution, saturated brine
solution, dried (MgSO.sub.4), concentrated in vacuo. Compound was
passed through a plug of silica to give 142 and used directly in
the next step without further purification.
27-b. Benzylidene Cleavage
[0184] To a solution of thioglycoside 142 [36.7 mmol] in a mixture
of MeCN/MeOH/H.sub.2O [2:1:0.1, 155 mL] was added
p-toluenesulphonic acid [200 mg]. The resulting reaction mixture
was stirred at 75.degree. C. for 2 hrs. The reaction was allowed to
cool, to room temperature, water was added [50 mL] and the volatile
solvents [MeCN and MeOH] removed in vacuo. The resulting suspension
was filtered and the collected solid washed further with water
followed by petroleum ether and then dried under vacuum to afford
pure 143 [98%]; [M+H].sup.+=480.3, (99.8% pure by ELSD); HPLC
Method A, Rt=3.80 mins.
27-c. Sialyl Protection of a 6-OH Group
[0185] To a suspension of the dial 143 [10 mmol] in pyridine [20
mL] was added imidazole [1 mmol] and the resulting reaction mixture
was then heated to 120.degree. C. At this time TBDPS-Cl [12 mmol]
was added in portions and the reaction was stirred for 1 hr at
120.degree. C. After this time further TBDPS-Cl [0.4 eq.] was added
and the reaction was allowed to stir for a further hour. The
reaction mixture was then cooled, and the volatiles removed in
vacuo. The residue was taken up in DCM and washed with 1 molar HCl
solution, dried (MgSO.sub.4) and the solvent removed in vacuo. The
residue was washed with petroleum ether to afford pure 144 as a
white solid [99%]; HPLC Method A Rt=6.66 mins (100% purity by
ELSD); [M+H].sup.+=718.57.
27-d. Formation of a 4-O-Triflate
[0186] To a solution of 144 [2 mmol] in DCM [20 mL] was added
pyridine [4 mmol] and the resulting mixture cooled to 0.degree. C.
At this time triflic anhydride [3.2 mmol] was slowly added and the
reaction mixture was then allowed to return to room temperature.
The reaction was allowed to stir for one hour at room temperature
at which time it was diluted with DCM and washed with a solution of
0.5 molar HCl, dried (MgSO.sub.4) and the solvent removed in vacuo
to afford pure 145 [100%]; HPLC Method A, Rt=7.63 mins;
[M+H.sup.+]=850.66.
27-e. Formation of an Axial Azido Derivative
[0187] To a solution of triflate 145 [1 mmol] in DMF was added
NaN.sub.3 [3 mmol] and the resulting reaction mixture was allowed
to stir at room temperature for 10 hrs. The reaction mixture was
concentrated in vacuo, and the residue washed with water followed
by petroleum ether. The solid was then dried to provide the product
146 [99%]; HPLC Method A, Rt=7.23 mins; [M+H.sup.+]=743.5.
27-f. Removal of a Benzoyl Group by Transesterification
[0188] Compound 147 was prepared according to the procedure
described in General Method 2 and purified by column chromatography
[30% ethyl acetate/petroleum ethers] to afford a white solid [83%];
HPLC Method A, RT=6.53 min; [M+H].sup.+=639.2.
27-g. Deprotection of the 2-Amino Group
[0189] To a solution of the sugar 147 [4.29 mmol] in DMF/MeOH [1:2,
45 mL] at room temperature was added hydrazine hydrate [0.52 mL].
The reaction mixture was stirred for two hours at which time it was
filtered and the filtered solid washed with methanol. The filtrates
were combined, the solvents removed in vacuo, residue taken up in
CHCl.sub.3, washed with saturated brine, dried (MgSO.sub.4), and
the solvent again removed in vacuo to provide a white solid 148
[88%]; HPLC Method A, Rt=5.96 mins; [M+H].sup.+=473.3.
27-h. Reprotection of the 2-Amino Group
[0190] To a solution of the sugar 148 [0.22 mmol] in MeOH [1.25 mL]
was added phthalic anhydride [0.4 mmol] and triethylamine [1 drop]
and the solution was allowed to stir overnight. The reaction
mixture was then concentrated in vacuo. The residue was the
dissolved in dry pyridine [0.25 mL], cooled to 0.degree. C., and
acetic anhydride [60 .mu.L] added dropwise. The reaction was
allowed to stir overnight. The reaction was then concentrated, the
residue taken up in CHCl.sub.3 and washed with 10% citric acid
solution, saturated sodium bicarbonate solution, saturated brine
solution, dried (MgSO.sub.4), the solvent removed in vacuo, and the
residue purified by column chromatography [20% ethyl
acetate/petroleum ethers] to afford the product as a white solid
149 [64%]; HPLC Method A, Rt=7.29 mins; [M+H].sup.+=645.35.
[0191] 27-i. Glycosylation to Form the O-Methyl Glycoside
[0192] To a solution of the sugar 149 [0.775 mmol] in DCM [5 mL]
was added 3 angstrom molecular sieves, MeOH [12 mmol] and finally
DMTST [2.32 mmol]. The reaction mixture was allowed to stir for 30
mins at which time the reaction was quenched with triethylamine
[2.37 mmol], filtered and the filtrate concentrated in vacuo. The
residue was taken up in DCM and washed with water, 10% citric acid
solution, saturated sodium hydrogen carbonate solution, saturated
brine, dried (MgSO.sub.4) and the solvent removed in vacuo to
provide a yellow oil. The oil was purified by column chromatography
[20% ethyl acetate/petroleum ethers] to provide the product as a
yellow oil [79%]; HPLC Method A, Rt=7.20 mins;
[M+Na].sup.+=651.3.
27-j. Zemplen Deprotection
[0193] Compound 150 was prepared according to the procedure
described in General Method 2 and was purified by column
chromatography [25% ethyl acetate/petroleum ethers] as a white
solid [67%]; HPLC Method A, Rt=6.88 mins; [M+Na].sup.+=609.7.
27-k. Formation of an O-Me Glycoside, Beta 1-3 Linked
Disaccharide
[0194] Donor 60 [0.128 mmol] and acceptor [85.2 mmol] were
dissolved 1,2-DCE [1.0 mL]. 4 Angstrom molecular sieves were added
and the mixture was stirred for 15 mins. TMSOTf [2.8 .mu.mol] was
then added and the reaction left to stir for 90 mins. The reaction
mixture was then quenched with triethylamine, diluted with
CHCl.sub.3, washed with saturated NaHCO.sub.3 solution, dried
(MgSO.sub.4) and the solvents removed in vacuo. The residue was
purified by column chromatography to afford 151 [20%]; HPLC Method
A, Rt=7.60 mins; [M+Na].sup.+=1226.67
EXAMPLE 28
Formation of Alternatively Linked Disaccharide Scaffolds
##STR00099##
[0195] 28-a. Glycosylation with a Trichloroacetimidate Donor to
Afford Disaccharides (153-a to 153-h)
[0196] To solutions of the acceptor molecules 152a-152h (1.8 mmol)
and the donor 2 (2.7 mmol) in dry 1,2-dichloroethane (84 mL) is
added 3 A acid-washed molecular pellets (4 g) and the resulting
mixture stirred for 20 min. To the mixture was then added Methyl
Triflate (1.8 mL of a 0.1M solution in dry 1,2-dichloroethane, 0.18
mmol) and the reaction then stirred for 30 mins. After this time
triethylamine (6 mL) was added and the suspension filtered, washed
with dichloromethane and all solvent removed in vacuo. This residue
was purified by column chromatography to yield the title compounds
as indicated in table 3 below.
TABLE-US-00005 TABLE 3 Disaccharide Products From Glycosylation
with Donor 2 Acceptor Product ##STR00100## ##STR00101##
##STR00102## ##STR00103## ##STR00104## ##STR00105## ##STR00106##
##STR00107## ##STR00108## ##STR00109## ##STR00110## ##STR00111##
##STR00112## ##STR00113## ##STR00114## ##STR00115##
##STR00116##
28-b. Azide Reduction and deprotection (154a-154h)
[0197] Compounds 153a-153h are hydrogenolysed at 60 psi for 1 hour
with catalytic 10% palladium on activated charcoal in ethanol, to
yield upon filtration and evaporation, the corresponding diamines
in which the benzylidene ring has also been cleaved as indicated in
the table below. These diamines may be used in their crude form for
further reactions.
##STR00117## ##STR00118##
28-c. Amide Formation-HBTU Coupling (155a-155h)
[0198] Compounds 155a(i)-155a(iv) to 155h(i)-155h(iv) are prepared
by reaction of the diamines 154a to 154h with carboxylic acids
according to the procedure described in General Method 4 in a
combinatorial manner. This every diamine may be reacted with an
excess of every carboxylic acid to produce the bis-amide
products.
TABLE-US-00006 TABLE 4 Acids i-iv Shown Below are Reacted with
Diamines to Give Products as Listed ##STR00119## ##STR00120##
##STR00121## ##STR00122## Acids Diamine i ii iii iv 154a 155a(i)
155a(ii) 155a(iii) 155a(vi) 154b 155b(i) 155b(ii) 155b(iii)
155b(vi) 154c1 155c1(i) 155c1(ii) 155c1(iii) 155c1(vi) 154c2
155c2(i) 155c2(ii) 155c2(iii) 155c2(vi) 154d 155d(i) 155d(ii)
155d(iii) 155d(vi) 154e 155e(i) 155e(ii) 155e(iii) 155e(vi) 145f
155f(i) 155f(ii) 155f (iii) 155f(vi) 154g 155g(i) 155g(ii)
155g(iii) 155g(vi) 154h 155h(i) 155h(ii) 155h(iii) 155h(vi)
Examples of Table 4 ##STR00123## ##STR00124##
28-d. Silyl Deprotection
[0199] The t-butyldiphenylsilyl groups are removed from Compounds
155 (as appropriate) by treatment of the silylated compound in
N,N-dimethylformamide with tetrabutylammonium fluoride, followed by
removal of the solvents in vacuo and purification by mass based
fractionation on a C18 HPLC column.
EXAMPLE 29
Formation of Alternatively Amide Linked Disaccharide Scaffolds
Reduction of an Azide to an Amine
[0200] Amino sugars can be obtained by reduction of corresponding
azido sugars according to the procedure described in General Method
3. Alternatively, the azide may be reduced selectively by
hydrogenolysis at atmospheric pressure over 5% palladium on
charcoal in methanol for 30 minutes. This latter method is suitable
for the reduction of azides in the presence of benzyl ethers.
Filtration of the solution and removal of the solvents in vacuo
yields the crude aminosugar suitable for further reaction.
Hydrogenolysis is also employed to remove the carbobenzyloxy group
from compound 152d (see Amines Used in Example 28).
Formation of an Anhydride and Reaction with an Amine
[0201] Anhydrides are formed according to the following general
method. Azaleic acid monomethyl ester, suberic acid monomethyl
ester or fumaric acid monoethyl ester [2 equivalents] are dissolved
in anhydrous dichloromethane to form a 10 milillolar solution. To
this solution is added diisopropylcarbodiimide [1 equivalent] and
triethylamine [1 equivalent] and the solution stirred at room
temperature for 45 minutes. After this time, the solution of acid
anhydride is evaporated, redissolved in N,N-dimethylformamide to
form a 10 millimolar solution and added to a solution of the crude
sugar amine (selected from Amines Used in Example 28) in
N,N-dimethylformamide. The reaction mixture is stirred for 1 hour.
The reaction mixture is quenched with water, acidified to pH 4 and
extracted with ethyl acetate and back extracted with 10% sodium
hydrogen carbonate solution to yield the crude sugar amide. The
solvents are removed in vacuo and the esters hydrolysed by the
addition of 5 equivalent of lithium hydroxide to an wet methanolic
solution of the crude ester. Acidification of this mixture followed
by removal of the solvents in vacuo yields the crude half acid
amide which is partially purified by passing through a short bed of
silica gel. This crude material in which all other protecting
esters have been cleaved is suitable for further reaction.
Compounds formed by this process are displayed in Half Acid Amides
formed in Example 29 below.
Formation of a Dimeric Derivative
[0202] The crude sugar half acid amide, is then dissolved in
N,N-dimethylformamide and treated with 1 equivalent of ethyl
diisopropylamine, 1 equivalent of HBTU and finally 1.3 equivalents
of the crude sugar amine. The reaction mixture is stirred for 30 to
60 minutes at room temperature, quenched by the addition of water
and solvents removed in vacuo The crude residue is finally purified
by mass based fractionation to furnish the desired bis amide linked
scaffolds. A combinatoral matrix of acid and amine results in a
wide diversity of bis amide linked scaffolds as exemplified in
Table 5.
Amines used in Example 29:
##STR00125##
Half Acid Amides formed in Example 29:
##STR00126##
TABLE-US-00007 TABLE 5 Products Resulting From Reaction of
Compounds From "Amines used in Example 29", with "Half Acid Amides
formed in Example 29" Amine Acid 13 156 50 59 152d 157A 157A13 157B
157B13 157C 157C13 158A 158A13 158A156 158B 158B13 158B156 158C
158C13 158C156 159A 159A13 159A156 159A50 159B 159B13 159B156
159B50 159C 159C13 159C156 159C50 160A 160A13 160A156 160A50 160A59
160B 160B13 160B156 160B50 160B59 160C 160C13 160C156 160C50 160C59
161A 161A13 161A156 161A50 161A59 161A152d 161B 161B13 161B156
161B50 161B59 161B152d 161C 161C13 161C156 161C50 161C59 161C152d
Example structures from Table 5: ##STR00127## ##STR00128##
EXAMPLE 30
Formation of Amide Linked Disaccharide Scaffolds
[0203] Glucuronic acid 14, is dissolved in N,N-dimethylformamide to
form a 10 millimolar solution. To this solution is added
triethylamine [1.1 equivalents] followed by HBTU [1.05 equivalents.
The mixture is stirred for 3 minutes at room temperature, after
which time a concentrated solution of the amine [20-30 millimolar;
1 equivalent], as prepared in Example 29 [amines 13, 156, 152d, 50
and 59] is rapidly added. The reaction mixture is stirred for a
further 45 minutes, then quenched with an equal volume of 10%
citric acid in water, and extracted with ethyl acetate. The organic
layers are dried over magnesium sulfate and solvents removed in
vacuo to yield the crude product which is further purified by
column chromatography, to yield the desired product.
Reaction Products:
##STR00129##
[0205] The Boc, isopropylidene and benzylidene protecting groups
may be removed by treatment with TFA according to general procedure
5, acetate and benzoate protecting groups are removed according to
general procedure 2.
EXAMPLE 31
Synthesis of an Alkylated 2-Deoxy-2-Amino Disaccharidic
Compound
##STR00130##
[0206] 31-a and 31-b. N-Alkylation
[0207] (31-a). To a solution of the sugar 109-03 [7.4 mg] in
THF/MeOH [84 .mu.L/9.4 .mu.L] was added benzaldehyde [0.71 .mu.L]
and the mixture stirred for 2 hrs at room temperature. To the
mixture was then added acetic acid [0.5 .mu.L] and NaCNBH.sub.3
[0.8 mg] and the reaction was allowed to stir overnight at room
temperature. The reaction was neutralised and concentrated in
vacuo. The residue was taken up in DCM and washed with a saturated
brine solution, dried (MgSO.sub.4) and the solvent removed in
vacuo. (31-b). The residue was treated with Ac.sub.2O/pyridine
[1:3] solution for two hours for the purpose of analysis; HPLC
Method A, Rt (168 monobenzylated-monoacetylated)=7.56 mins, (169
bis-benzylated)=8.77 mins; mono-benzylated-mono-acetylated
[M+H].sup.+=1391.9, bis-benzylated [M+H].sup.+=1339.9.
EXAMPLE 32
Synthesis of Benzimidazole Compounds
##STR00131##
[0208] 32-a. Fluorine Displacement
[0209] A solution of 3-fluoro-2-nitro-trifluoromethylbenzene
[0.0715 mmol], triethylamine [0.0861 mmol] in DMF [250 .mu.L] was
added to a flask containing diamine 68 [0.0241 mmol]. The resulting
reaction mixture was then stirred at 50.degree. C. for 16 hrs. At
this time the reaction was allowed to cool and the product was
purified by preparative TLC [mobile phase ethyl acetate]. The
product was collected in quantitative yield; HPLC Method A, Rt=7.51
mins; [M+Na].sup.+=1230.6
32-b. Formation of Benzimidazole
[0210] A solution of SnCl.sub.2 [300 .mu.L of SnCl.sub.2.H.sub.2O
at 0.32 molar in DMF] was added to a flask containing compound 170
[2.48 .mu.mol]. The reaction mixture was then stirred at 80.degree.
C. for 16 hrs. The reaction mixture was diluted with EtOAc/H.sub.2O
[1:1, 5 mL], filtered through a pad of celite. The filtrated was
separated into aqueous and organic layers and the organic layer
washed with H.sub.2O, dried (MgSO.sub.4) and the solvent removed in
vacuo to afford a mixture of products 171 and 172; HPLC Method A,
Rt (171)=7.18 mins, Rt (172)=7.41 mins; [M+H].sup.+ (171)=1186.8,
[M+H].sup.+ (172)=11.58.8.
EXAMPLE 33
Synthesis of a N-Acetyl-lactosamine Based Library of 6'-Hydroxy
Phosphonates
##STR00132##
[0212] Reactions were carried out in identical series for resins
174-1 and series 174-2. After step 33). After step 33-b each series
was divided into 12 portions for the individual alkylations.
Alkylating Agents for Example 33, where R=Sulphonate or
R=Halide.
##STR00133##
33-a. Glycosylation
[0213] Resin [0.47 mmol] was weighed into a reactor and molecular
sieves [200 mg], thioglycoside donor sugar 174 [2.35 mmol] and
dichloromethane [.about.1.5 mL] was added. To the mixture was then
added DMTST [2.35 mmol]. The reaction vessel was sealed, shaken and
reacted for 5 hours. At this time the reaction was then quenched by
the addition of triethylamine, and the molecular sieves removed
from the resin. The resin was then washed with DMF, MeOH/CHCl.sub.3
(1:1) and dichloromethane. The resin was then dried under
vacuum.
33-b. Solid Phase Silylether Deprotection
[0214] A solution of PSHF (proton sponge hydrogen fluoride) (0.5
Molar in DMF/Acetic Acid, 95:5) was prepared. The resins [1.41
mmol] was added to the solution and the reaction was stirred at
65.degree. C. for 24 hours. The rein was then washed with DMF,
MeOH/CH.sub.3COOH/THF, 1:1:8, THF and DCM, and then dried under
high vacuum.
33-c. Solid Phase Alkylation
[0215] Resins 176 [0.047 mmol] were individually reacted with a
0.25 molar solution of tert-butoxide in DMF (5 min) and then an
alkylating agent (see above), [0.25 molar of alkylating agent in
DMF, 20 min] was reacted with the resin. The resins were washed
with DMF and again treated with the two solutions, this procedure
was repeated a further four times. The final wash of the resins was
performed as above; with DMF, THF/MeOH/CH.sub.3CO.sub.2H (8:1:1),
THF, DCM and MeOH. The resins were then dried overnight.
33-d. Cleavage of Disaccharide from Resin
[0216] The resins 177 [0.047 mmol] were separately treated with a
7% hydrazine hydrate/DMF solution [2 mL] overnight. The resin was
filtered and the resin washed with DMF. The filtrates were combined
and the solvent removed in vacuo. The residue was taken up in DCM
and washed with water and saturated brine solution, dried
(MgSO.sub.4) and the solvent removed in vacuo. The residue was then
treated with a solution of Ac.sub.2O/pyridine [1 mL, 1:3] for three
hours. The Solvents were removed in vacuo and the product purified
by column chromatography.
33-e. Removal of the Pivaloyl Protecting Group
[0217] To a solution of NaOMe/MeOH/THF [2 mL, .about.2 molar] was
added the pivaloyl protected disaccharide 178 [0.03 mmol]. The
reaction mixture was heated at reflux until TLC indicated the
reaction was complete. At completion, reaction mixture pH was
reduced to .about.5 with amberlite IR-120-H.sup.+ resin. The
reaction was filtered, and the solvent concentrated in vacuo
33-f. Deprotection of Hydroxyphosphonates and Benzyl Ether
Cleavage
[0218] Compound 178 (after 33-e) [0.0193 mmol] was dissolved in dry
dichloromethane [2 mL] under a nitrogen atmosphere, the solution
cooled to 0.degree. C. and trimethylsilyl bromide [0.097 mmol] was
added. After stirring at 0.degree. C. for 30 mins a solution of
ammonia in methanol [12 .mu.L of 28% aq ammonia in 20 mL methanol]
was added. The solvents were removed to give the crude free
hydroxyphosphonate as an ammonium salt. Final products were
purified by mass fractioning HPLC.
TABLE-US-00008 TABLE 6 Final products Synthesised in Example 33.
##STR00134## Comp. No. R R1 R2 Note 179a Ra H OMe 179b Rb H OMe
179c Rc H OMe 179d Rd H OMe 179e Re H OMe 179f Rf H OMe 179g Rg H
OMe 179h Rh H OMe 179i Ri H OMe 179j Rj H OMe 179k Rk H OMe 179l Rl
H OMe 179m Ra H,OH OH,H Anomeric lactol 179n Rb H,OH OH,H Anomeric
lactol 179o Rc H,OH OH,H Anomeric lactol 179p Rd H,OH OH,H Anomeric
lactol 179q Re H,OH OH,H Anomeric lactol 179r Rf H,OH OH,H Anomeric
lactol 179s Rg H,OH OH,H Anomeric lactol 179t Rh H,OH OH,H Anomeric
lactol 179u Ri H,OH OH,H Anomeric lactol 179v Rj H,OH OH,H Anomeric
lactol 179w Rk H,OH OH,H Anomeric lactol 179x Rl H,OH OH,H Anomeric
lactol Side Arms for Table 6 ##STR00135## ##STR00136## ##STR00137##
##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142##
##STR00143## ##STR00144## ##STR00145## ##STR00146##
EXAMPLE 34
Preparation of Guanidine
##STR00147##
[0219] 34-a. Reaction Conditions to form a Thiourea
[0220] Compounds 111-14 and 180 were prepared as a mixture
(unpurified) by reaction of compound 110-20 according to the
procedure described in General Method 12. The mixture was used
directly in the next step.
34-b. Formation of a Guanidine
[0221] The sugar mixture (111-14 and 180) (0.025 mmol) was
dissolved in methanol (0.5 mL) and concentrated aqueous ammonium
hydroxide (0.5 ml) was added. The reaction was stirred at room
temperature for 4 hours. The solvents were then removed in vacuo
and the residue purified by LCMS.
[0222] In a cognate manner, benzylamine, ethylamine, and other
primary or secondary amines can be substituted for ammonia to yield
the corresponding substituted guanidiniums. Products are shown in
table 7.
TABLE-US-00009 TABLE 7 Guanidinium products ##STR00148## Prod-
Starting Synth. Rt Yield uct material method R5* R2 R3 R4 M + H
(mins) (%) 180 110-20 12 H C A I 937.5 6.31 49 181 111-31 17 H C A
I 800.37 5.23 75 182 110-20 34-b Bn C A I ND ND ND 183 110-20 34-b
Et C A I ND ND ND 184 110-20 34-b Me C A I ND ND ND R5*
substituents are Hydrogen (H), Benzyl (Bn), Ethyl (Et), or Methyl
(Me): Substituents R2-R4 are as found in Table 2, Example 24.
[0223] I. K. C. Nicolaou; J. M. Salvino, K. Raynor; S. Pietranico;
T. Reisine; R. M. Freidinger, R. Hirschmann, Pept.: Chem., Struct.
Biol., Proc. Am. Pept. Symp., 11.sup.th, 1990 [0224] II. (a) H.
Kunz, T. Wundberg, C. Kallus, T. Opatz, S. Henke, W. Schmidt,
Angew. Chem. Int. Ed., 1998, 37, No. 18, (b) K. Kallus, T.
Wundberg, W. Schmidt, S. Henke, H. Kunz, Tet. Lett., 40, 1999,
7783-7786, (c) U. Hunger, T. Maidhof, O. Knoll, H. Kunz, Poster
Presentation, 20.sup.th International Carbohydrate Symposium,
Hamburg-Germany, (d) T. Opatz, C. Kallus, T. Wundberg, W. Schmidt,
S. Henke, H. Kunz, Poster Presentation, 20.sup.th International
Carbohydrate Symposium, Hamburg-Germany. [0225] III. R. Hirschmann,
K. C. Nicolaou, S. Pietramico, J. Salvino, E. M. Lealy, W. C.
Shakepeare, P. S. Spengler, P. Hamley, A. B. Smith, T. Reisine, K.
Raynor, C. Donaldson, W. Vale, L. Maechler, R. M. Freidinger, C. D.
Strader, J. Am. Chem. Soc., 1993, 115, 12550
* * * * *