U.S. patent application number 10/507829 was filed with the patent office on 2005-11-10 for building block forming a c-c or a c-hetero atom bond uponreaction.
This patent application is currently assigned to NUEVOLUTION A/S. Invention is credited to Felding, Jakob, Godskesen, Michael Anders, Gouliaev, Alex Haahr, Jensen, Kim Birkebaek, Lundorf, Mikkel Dybro, Pedersen, Henrik, Sams, Christian.
Application Number | 20050247001 10/507829 |
Document ID | / |
Family ID | 28046589 |
Filed Date | 2005-11-10 |
United States Patent
Application |
20050247001 |
Kind Code |
A1 |
Gouliaev, Alex Haahr ; et
al. |
November 10, 2005 |
Building block forming a c-c or a c-hetero atom bond
uponreaction
Abstract
A building block having the dual capabilities of transferring
genetic information and functional entity precursor to a recipient
reactive group is disclosed. The building block may be used in the
generation of a single complex or libraries of different complexes,
wherein the complex comprises an encoded molecule linked to an
encoding element. Libraries of complexes are useful in the quest
for pharmaceutically active compounds.
Inventors: |
Gouliaev, Alex Haahr;
(Veksoe Sjaelland, DK) ; Pedersen, Henrik;
(Bagsvaerd, DE) ; Jensen, Kim Birkebaek;
(Roedovre, DK) ; Lundorf, Mikkel Dybro;
(Copenhagen, DK) ; Sams, Christian; (Frederiksberg
C, DK) ; Felding, Jakob; (Charlottenlund, DK)
; Godskesen, Michael Anders; (Vedbaek, DK) |
Correspondence
Address: |
BROWDY AND NEIMARK, P.L.L.C.
624 NINTH STREET, NW
SUITE 300
WASHINGTON
DC
20001-5303
US
|
Assignee: |
NUEVOLUTION A/S
COPENHAGEN
DK
|
Family ID: |
28046589 |
Appl. No.: |
10/507829 |
Filed: |
May 11, 2005 |
PCT Filed: |
March 14, 2003 |
PCT NO: |
PCT/DK03/00176 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60364056 |
Mar 15, 2002 |
|
|
|
60434423 |
Dec 19, 2002 |
|
|
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Current U.S.
Class: |
52/596 |
Current CPC
Class: |
C12N 15/1034 20130101;
C07D 405/04 20130101; C40B 40/00 20130101; C07H 21/00 20130101;
C12N 15/1068 20130101 |
Class at
Publication: |
052/596 |
International
Class: |
E04C 002/04; E04B
005/04 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 15, 2002 |
DK |
PA 2002 0415 |
Dec 19, 2002 |
DK |
PA 2002 01950 |
Claims
1. A building block of the general formulaComplementing
Element-Linker-Carrier-C--F-connecting group--Functional entity
precursorcapable of transferring a Functional entity precursor to a
recipient reactive group, wherein Complementing Element is a group
identifying the Functional entity precursor, Linker is a chemical
moiety comprising a spacer and a S--C-connecting group, wherein the
spacer is a valence bond or a group distancing the Functional
entity precursor to be transferred from the complementing element
and the S--C-connecting group connects the spacer with the Carrier,
Carrier is arylene, heteroarylene, C.sub.1-C.sub.6 alkylene,
C.sub.1-C.sub.6 alkenylene, C.sub.1-C.sub.6 alkynylene, or
--(CF.sub.2).sub.m-- substituted with 0-3 R.sup.1 wherein m is an
integer between 1 and 10; R.sup.1 are independently selected from
the group consisting of --H, --OR.sup.2, --NR.sup.2.sub.2,
-Halogen, --NO.sub.2, --CN, --C(Halogen).sub.3, --C(O)R.sup.2,
--C(O)NHR.sup.2, C(O)NR.sup.2.sub.2, --NC(O)R.sup.2,
--S(O).sub.2NHR.sup.2, --S(O).sub.2NR.sup.2, --S(O).sub.2R.sup.2,
--P(O).sub.2--R.sup.2, --P(O)--R.sup.2, --S(O)--R.sup.2,
P(O)--OR.sup.2, --S(O)--OR.sup.2, and --N.sup.+R.sup.2.sub.3,
wherein R.sup.2 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, or aryl, C--F-connecting group is
selected from the group consisting of --SO.sub.2--O--,
--O--SO.sub.2--O--, --C(O)--O--, --S.sup.+(R.sup.3)--,
--C--U--C(V)--O--, --P.sup.+(W).sub.2--O--, and --P(W)--O--, where
U is --C(R.sup.2).sub.2--, --NR.sup.2-- or --O--; V is .dbd.O or
.dbd.NR.sup.2 and W is --OR.sup.2 or --N(R.sup.2).sub.2, Functional
entity precursor is --C(H)(R.sup.3)--R.sup.4 or functional entity
precursor is heteroaryl or aryl optionally substituted with one or
more substituents belonging to the group comprising R.sup.3 and
R.sup.4, Wherein R.sup.3 and R.sup.4 independently is H, alkyl,
alkenyl, alkynyl, alkadienyl, cycloalkyl, cycloheteroalkyl, aryl or
heteroaryl, optionally substituted with one or more substituents
selected from the group consisting of SnR.sup.5R.sup.6R.sup.7,
Sn(OR.sup.5)R.sup.6R.sup.7, Sn(OR.sup.5)(OR.sup.6)R.sup.7,
BR.sup.5R.sup.6, B(OR.sup.5)R.sup.6, B(OR.sup.5)(OR.sup.6),
halogen, CN, CNO, C(halogen) .sub.3, OR.sup.5, OC(.dbd.O) R.sup.5,
OC(.dbd.O)OR.sup.5, OC (.dbd.O) NR.sup.5R.sup.6, SR.sup.5,
S(.dbd.O) R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, N.sub.3, NR.sup.5R.sup.6,
N.sup.+R.sup.5R.sup.6R.sup.7, NR.sup.5OR.sup.6, NR.sup.5N
R.sup.6R.sup.7, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, NC,
P(.dbd.O)(OR.sup.5)OR.sup.6, P.sup.+R.sup.5R.sup.6R.sup.7,
C(.dbd.O) R.sup.5, C(.dbd.NR.sup.5)R.sup.6,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.NNR.sup.6), C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6,
C(.dbd.O)NR.sup.5NR.sup.6R.sup.7, C(.dbd.NR.sup.5)NR.sup.6R.sup.7,
C(.dbd.NOR.sup.5)NR.sup.6R.sup.7 and R.sup.8, wherein, R.sup.5,
R.sup.6, and R.sup.7 independently is H, alkyl, alkenyl, alkynyl,
alkadienyl, cycloalkyl, cycloheteroalkyl, aryl or heteroaryl,
optionally substituted with one or more substituents selected from
the group consisting of halogen, CN, CNO, C(halogen).sub.3, .dbd.O,
OR.sup.8, OC(.dbd.O)R.sup.8, OC(.dbd.O)OR.sup.8,
OC(.dbd.O)NR.sup.8R.sup.9, SR.sup.8, S(.dbd.O)R.sup.8,
S(.dbd.O).sub.2R.sup.8, S(.dbd.O).sub.2NR.sup.8R.sup.9, NO.sub.2,
N.sub.3, NR.sup.8R.sup.9, N.sup.+R.sup.8R.sup.9R.sup.10,
NR.sup.5OR.sup.6, NR.sup.5NR.sup.6R.sup.7,
NR.sup.8C(.dbd.O)R.sup.9, NR.sup.8C(.dbd.O)OR.sup.9,
NR.sup.8C(.dbd.O)NR.sup.9R.sup.10, NC, P(.dbd.O)(OR.sup.8)OR.sup.9,
P.sup.+R.sup.5R.sup.6R.sup.7, C(.dbd.O)R.sup.8,
C(.dbd.NR.sup.8)R.sup.9, C(.dbd.NOR.sup.8)R.sup.9,
C(.dbd.NNR.sup.8R.sup.9), C(.dbd.O)OR.sup.8,
C(.dbd.O)NR.sup.8R.sup.9,
C(.dbd.O)NR.sup.8OR.sup.9C(.dbd.NR.sup.5)NR.sup.6R.sup.7,
C(.dbd.NOR.sup.5)NR.sup.6R.sup.7 or
C(.dbd.O)NR.sup.8NR.sup.9R.sup.10, wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring, wherein, R.sup.8, R.sup.9, and R.sup.10 independently is H,
alkyl, alkenyl, alkynyl, alkadienyl, cycloalkyl, cycloheteroalkyl,
aryl or heteroaryl and wherein R.sup.8 and R.sup.9 may together
form a 3-8 membered heterocyclic ring or R.sup.8 and R.sup.10 may
together form a 3-8 membered heterocyclic ring or R.sup.9 and
R.sup.10 may together form a 3-8 membered heterocyclic ring.
2. A compound according to claim 1 wherein, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11' or functional entity
precursor is heteroaryl or aryl substituted with 0-3 R.sup.11, 0-3
R.sup.13 and 0-3 R.sup.15, wherein R.sup.11 and R.sup.11'0 are
independently H, or selected from the group consisting of a
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.4-C.sub.8 alkadienyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl, and heteroaryl, said group
being substituted with 0-3 R.sup.12, 0-3 R.sup.13 and 0-3 R.sup.15,
or R.sup.11 and R.sup.11' are C.sub.1-C.sub.3
alkylene-NR.sup.12.sub.2, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)R.sup.16, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)OR.sup.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12.su- p.2, C.sub.1-C.sub.2 alkylene-O--N
R.sup.12C(O)R.sup.16, or C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)OR.sup.16 substituted with 0-3 R.sup.15,
where R.sup.12 is H or selected independently from the group
consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl, and heteroaryl, said group
being substituted with 0-3 R.sup.13 and 0-3 R.sup.15, R.sup.13 is
selected independently from the group consisting of --N.sub.3,
--CNO, --C(NOH)NH.sub.2, --NHOH, --NHNHR.sup.17, --C(O)R.sup.17,
--SnR.sup.17.sub.3, --B(OR.sup.17).sub.2, and
--P(O)(OR.sup.17).sub.2 or the group consisting of C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, and C.sub.4-C.sub.8 alkadienyl,
said group being substituted with 0-2 R.sup.14, where R.sup.14 is
independently selected from the group consisting of --NO.sub.2,
--C(O)OR.sup.17, --COR.sup.17, --CN, --OSiR.sup.17.sub.3,
--OR.sup.17 and --NR.sup.17.sub.2; R.sup.15 is .dbd.O, --F, --Cl,
--Br, --I, --CN, --NO.sub.2, --OR.sup.17, --NR.sup.17.sub.2,
--NR.sup.17--C(O)R.sup.16, --NR.sup.17--C(O)OR.sup.17, --SR.sup.17,
--S(O)R , --S(O).sub.2R.sup.17, --COOR.sup.17,
--C(O)NR.sup.17.sub.2 or --S(O).sub.2NR.sup.17.sub.2, R.sup.16 is
H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.7 cycloalkyl, aryl or C.sub.1-C.sub.6
alkylene-aryl substituted with 0-3 substituents independently
selected from --F, --Cl, --NO.sub.2, --R.sup.2, --OR ,
--SiR.sup.2.sub.3; R.sup.17 is selected independently from the
group consisting of H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, aryl, C.sub.1-C.sub.6 alkylene-aryl, 24G is H or
C.sub.1-C.sub.6 alkyl and n is 1,2,3 or 4.
3. A compound according to claim 2 wherein, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11' or functional entity
precursor is heteroaryl or aryl substituted with 0-3 R.sup.11, 0-3
R.sup.13 and 0-3 R.sup.15, wherein R.sup.11 and R.sup.11'0 are
independently H, or selected from the group consisting of a
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.4-C.sub.8 alkadienyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl, and heteroaryl, said group
being substituted with 0-3 R.sup.12, 0-3 R.sup.13 and 0-3 R.sup.15,
or R.sup.11 and R.sup.11'0 are C.sub.1-C.sub.3
alkylene-NR.sup.12.sub.2, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)R.sup.16, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)OR.sup.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12.su- b.2, C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)R.sup.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)OR.sup.16 substituted with 0-3 R.sup.15,
where R.sup.12 is H or selected from the group consisting of
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl, and heteroaryl, said group being
substituted with 0-3 R.sup.13 and 0-3 R.sup.15, R.sup.13 is
selected from the group consisting of --N.sub.3, --CNO,
--C(NOH)NH.sub.2, --NHOH, --NHNHR.sup.17, --C(O)R.sup.17,
--SnR.sup.17.sub.3, --B(OR.sup.17).sub.2, and
--P(O)(OR.sup.17).sub.2 or the group consisting of C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, and C.sub.4-C.sub.8 alkadienyl,
said group being substituted with 0-2 R.sup.14, where R.sup.14 is
selected from the group consisting of --NO.sub.2, --C(O)OR.sup.17,
--COR.sup.17, --CN, --OSiR.sup.17.sub.3, --OR.sup.17 and
--NR.sup.17.sub.2; R.sup.15 is .dbd.O, --F, --Cl, --Br, --I, --CN,
--NO.sub.2, --OR.sup.17, --NR.sup.17.sub.2,
--NR.sup.17--C(O)R.sup.16, --NR.sup.17--C(O)OR.sup.16, --SR.sup.17,
--S(O)R.sup.17, --S(O).sub.2R.sup.17, --COOR.sup.17,
--C(O)NR.sup.17.sub.2 or --S(O).sub.2NR.sup.17.sub.2, R.sup.16 is
H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.7 cycloalkyl, aryl or C.sub.1-C.sub.6
alkylene-aryl substituted with 0-3 substituents independently
selected from the group consisting of --F, --Cl, --NO.sub.2,
--R.sup.2, --OR.sup.2, and --SiR.sup.2.sub.3; wherein R.sup.17 is
selected independently from the group consisting of H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, aryl, and
C.sub.1-C.sub.6 alkylene-aryl.
4. A compound according to claim 1 wherein, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11'0 wherein R.sup.11 and
R.sup.11' are C.sub.1-C.sub.3 alkylene-NR.sup.12.sub.2,
C.sub.1-C.sub.3 alkylene-NR.sup.12C(O)R.sup.16, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)OR.- sup.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12.sub.2, C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)R.sup.16, or C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)OR.sup.16 substituted with 0-3
R.sup.15.
5. A compound according to claim 1 wherein, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11' wherein R.sup.11 and
R.sup.11' are independently H, or selected from the group
consisting of a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.4-C.sub.8 alkadienyl,
C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7 cycloheteroalkyl, aryl,
and heteroaryl, said group being substituted with 0-3 R.sup.12, 0-3
R.sup.13 and 0-3 R.sup.15.
6. A compound according to claim 2 wherein, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11' wherein R.sup.11 and
R.sup.11' are independently H, or selected from the group
consisting of a C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl, and heteroaryl, said group
being substituted with 0-3 R.sup.12 and 0-3 R.sup.15, where
R.sup.12 is H or selected from the group consisting of
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl, and heteroaryl, R.sup.15 is .dbd.O, --F,
--Cl, --Br, --I, --CN, --NO.sub.2, --OR.sup.17, --NR.sup.17.sub.2,
--NR.sup.17--C(O)R.sup.16, --NR.sup.17--C(O)OR.sup.16, --SR.sup.17,
--S(O)R.sup.17, --S(O).sub.2R.sup.17, --COOR.sup.17,
--C(O)NR.sup.17.sub.2 or --S(O).sub.2NR.sup.17.sub.2, R.sup.17 is
selected from the group consisting of H, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.7 cycloalkyl, and C.sub.1-C.sub.6 alkylene-aryl.
7. A compound according to claim 1 wherein, Functional entity
precursor is heteroaryl or aryl substituted with 0-3 R.sup.11, 0-3
R.sup.13 and 0-3 R.sup.15.
8. A compound according to claim 2 wherein C--F-connecting group is
selected from the group consisting of --SO.sub.2--O--,
--O--SO.sub.2--O--, --C(O)--O--, --S.sup.+(R.sup.11)--,
--C--U--C(V)--O--, --P.sup.+(W).sub.2--O--, and --P(W)--O-- where U
is --C(R.sup.2).sub.2--, --NR.sup.2-- or --O--; V is .dbd.O or
.dbd.NR.sup.2 and W is --OR.sup.2 or --N(R.sup.2).sub.2.
9. A compound according to claim 2 wherein C--F-connecting group is
--S.sup.+(R.sup.11)--.
10. A compound according to claim 1 wherein C--F-connecting group
is selected from the group consisting of --SO.sub.2--O--,
--O--SO.sub.2--O--, --C(O)--O--, --S.sup.+(R.sup.17)--,
--C--U--C(V)--O--, --P.sup.+(W).sub.2--O--, and --P(W)--O-- where U
is --C(R.sup.2).sub.2--, --NR.sup.2-- or --O--; V is .dbd.O or
.dbd.NR.sup.2 and W is --OR.sup.2 or --N(R.sup.2).sub.2, wherein
R.sup.17 is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl,
aryl, or C.sub.1-C.sub.6 alkylene-aryl.
11. A compound according to wherein C--F-connecting group is chosen
from the group consisting of --SO.sub.2--O--, and
--S.sup.+(R.sup.17)--; wherein R.sup.17 is H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, aryl, or C.sub.1-C.sub.6
alkylene-aryl.
12. A compound according to claim 1 wherein, Spacer is a valence
bond, C.sub.1-C.sub.6 alkylene-A--, C.sub.2-C.sub.6 alkenylene-A--,
C.sub.2-C.sub.6 alkynylene-A--, or 25said spacer optionally being
connected through A to a linker selected from
--(CH.sub.2).sub.n--B--, 26and
--(CH.sub.2).sub.n--S--S--(CH.sub.2).sub.m--B--where A is a valence
bond, --C(O)NR.sup.17--, --NR.sup.17--, --O--, --S--, or
--C(O)--O--; B is a valence bond, --O--, --S--, --NR.sup.17-- or
--C(O)NR.sup.17-- and connects to S--C-connecting group; and n and
m independently are integers ranging from 1 to 10; and R.sup.17 is
H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, aryl, or
C.sub.1-C.sub.6 alkylene-aryl.
13. A compound according to claim 1 wherein, Spacer is a valence
bond, C.sub.1-C.sub.6 alkylene-A--, C.sub.2-C.sub.6 alkenylene-A--,
C.sub.2-C.sub.6 alkynylene-A--, or 27said spacer optionally being
connected through A to a linker selected from
--(CH.sub.2).sub.n--B--, 28and where A is a valence bond,
--C(O)NR.sup.17--, --NR.sup.17--, --S--, or --C(O)--O--; B is
--O--, --S--, --NR.sup.17--, or --C(O)NR.sup.17-- and connects to
S--C-connecting group; and n and m independently are integers
ranging from 1 to 6; and R.sup.17 is H, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.7 cycloalkyl, aryl, or C.sub.1-C.sub.6
alkylene-aryl.
14. A compound according to claim 1 wherein, S--C-connecting group
is a valence bond, --NH--C(.dbd.O)--, 29
15. A compound according to claim 2 wherein, the carrier is
selected from the group consisting of arylene, heteroarylene and
--(CF.sub.2).sub.m-- substituted with 0-3 R.sup.1 wherein m is an
integer between 1 and 10, and C--F-connecting group is
--SO.sub.2--O--, and the functional entity precursor is
--C(H)(R.sup.11)--R.sup.11'.
16. A compound according to claim 1 wherein, the carrier is
--(CF.sub.2).sub.m-- wherein m is an integer between 1 and 10, the
C--F-connecting group is --SO.sub.2--O--; and the functional entity
precursor is aryl or heteroaryl substituted with 0-3 R.sup.11, 0-3
R.sup.13 and 0-3 R.sup.15.
17. A compound according to claim 1 wherein Complementing element
is a nucleic acid.
18. A compound according to claim 1 where Complementing element is
a sequence of nucleotides selected from the group consisting of
DNA, RNA, LNA PNA, and morpholino derivatives.
19. A library of compounds according to claim 1, wherein each
different member of the library comprises a complementing element
having a unique sequence of nucleotides, which identifies the
functional entity.
20. A method for transferring a functional entity precursor to a
recipient reactive group, comprising the steps of providing one or
more building blocks according to claim 1, contacting the one or
more building blocks with a corresponding encoding element
associated with a recipient reactive group under conditions which
allow for a recognition between the one or more complementing
elements and the encoding elements, said contacting being performed
prior to, simultaneously with, or subsequent to a transfer of the
functional entity precursor to the recipient reactive group.
21. The method according to claim 20, wherein the encoding element
comprises one or more encoding sequences comprised of 1 to 50
nucleotides and the one or more complementing elements comprises a
sequence of nucleotides complementary to one or more of the
encoding sequences.
22. The method of claim 20, wherein the recipient reactive group is
a nucleophilic S- or N-atom, which may be part of a chemical
scaffold, and the activating catalyst is contains palladium.
Description
TECHNICAL FIELD OF THE INVENTION
[0001] The present invention relates to a building block comprising
a complementing element and a precursor for a functional entity.
The building block is designed to transfer the functional entity
precursor with an adjustable efficiency to a recipient reactive
group upon recognition between the complementing element and an
encoding element associated with the reactive group. The invention
also relates to a method for transferring a functional entity
precursor to recipient a reactive group.
BACKGROUND
[0002] The transfer of a chemical entity from one mono-, di- or
oligonucleotide to another has been considered in the prior art.
Thus, N. M. Chung et al. (Biochim. Biophys. Acta, 1971,
228,536-543) used a poly(U) template to catalyse the transfer of an
acetyl group from 3'-O-acetyladenosine to the 5'-OH of adenosine.
The reverse transfer, i.e. the transfer of the acetyl group from a
5'-O-acetyladenosine to a 3'-OH group of another adenosine, was
also demonstrated.
[0003] Walder et al. Proc. Natl. Acad. Sci. USA, 1979, 76, 51-55
suggest a synthetic procedure for peptide synthesis. The synthesis
involves the transfer of nascent immobilized polypeptide attached
to an oligonucleotide strand to a precursor amino acid attached to
an oligonucleotide. The transfer comprises the chemical attack of
the amino group of the amino acid precursor on the substitution
labile peptidyl ester, which in turn results in an acyl transfer.
It is suggested to attach the amino acid precursor to the 5'0 end
of an oligonucleotide with a thiol ester linkage.
[0004] The transfer of a peptide from one oligonucleotide to
another using a template is disclosed in Bruick RK et al. Chemistry
& Biology, 1996, 3:49-56. The carboxy terminal of the peptide
is initially converted to a thioester group and subsequently
transformed to an activated thioester upon incubation with Ellman's
reagent. The activated thioester is reacted with a first oligo,
which is 5'-thiol-terminated, resulting in the formation of a
thio-ester linked intermediate. The first oligonucleotide and a
second oligonucleotide having a 3'0 amino group is aligned on a
template such that the thioester group and the amino group are
positioned in close proximity and a transfer is effected resulting
in a coupling of the peptide to the second oligonucleotide through
an amide bond.
SUMMARY OF THE INVENTION
[0005] The present invention relates to a building block of the
general formula:
Complementing Element-Linker-Carrier-C--F-connecting
group-Functional entity precursor
[0006] capable of transferring a Functional entity precursor to a
recipient reactive group, wherein
[0007] Complementing Element is a group identifying the Functional
entity precursor,
[0008] Linker is a chemical moiety comprising a spacer and a
S--C-connecting group, wherein the spacer is a valence bond or a
group distancing the Functional entity precursor to be transferred
from the complementing element and the S--C-connecting group
connects the spacer with the Carrier
[0009] Carrier is arylene, heteroarylene, C.sub.1-C.sub.6 alkylene,
C.sub.1-C.sub.6 alkenylene, C.sub.1-C.sub.6 alkynylene, or
--(CF.sub.2).sub.m-- substituted with 0-3 R.sup.1 wherein m is an
integer between 1 and 10;
[0010] R.sup.1are independently selected from --H, --OR.sup.2,
--NR.sup.2.sub.2, --Halogen, --NO.sub.2, --CN, --C(Halogen).sub.3,
--C(O)R.sup.2, --C(O)NHR.sup.2, C(O)NR.sup.2.sub.2, --NC(O)R.sup.2,
--S(O).sub.2NHR.sup.2, --S(O).sub.2NR.sup.2.sub.2,
--S(O).sub.2R.sup.2, --P(O).sub.2--R.sup.2, --P(O)--R.sup.2,
--S(O)--R.sup.2, P(O)--OR.sup.2, --S(O)--OR.sup.2,
--N.sup.+R.sup.2.sub.3, wherein R.sup.2 is H, C.sub.1-C6 alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6 alkynyl, or aryl,
[0011] C--F-connecting group is chosen from the group consisting of
--SO.sub.2--O--, --O--SO.sub.2--O--, --C(O)--O--,
--S.sup.+(R.sup.3RRrr)-- -, --C--U--C(V)--O--,
--P.sup.+(W).sub.2--O--, --P(W)--O-- where U is --C(R.sup.2).sub.2,
--NR.sup.2-- or --O--; V is .dbd.O or .dbd.NR.sup.2 and W is
--OR.sup.2 or --N(R.sup.2).sub.2
[0012] Functional entity precursor is --C(H)(R.sup.3)--R.sup.4 or
functional entity precursor is heteroaryl or aryl optionally
substituted with one or more substituents belonging to the group
comprising R.sup.3 and R.sup.4.
[0013] Wherein R.sup.3 and R.sup.4 independently is H, alkyl,
alkenyl, alkynyl, alkadienyl, cycloalkyl, cycloheteroalkyl, aryl or
heteroaryl, optionally substituted with one or more substituents
selected from the group consisting of SnR.sup.5R.sup.6R.sup.7,
Sn(OR.sup.5)R.sup.6R.sup.7, Sn(OR.sup.5)(OR.sup.6)R.sup.7,
BR.sup.5R.sup.6, B(OR.sup.5)R.sup.6, B(OR.sup.5)(OR.sup.6),
halogen, CN, CNO, C(halogen).sub.3, OR.sup.5, OC(.dbd.O)R.sup.5,
OC(.dbd.O)OR.sup.5, OC(.dbd.O)NR.sup.5R.sup.6, SR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, N.sub.3, NR.sup.5R.sup.6,
N.sup.+R.sup.5R.sup.6R.sup.7, NR.sup.5OR.sup.6,
NR.sup.5NR.sup.6R.sup.7, NR.sup.5C(.dbd.O)R.sup.6
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, NC,
P(.dbd.O)(OR.sup.5)OR.sup.6, P.sup.+R.sup.5R.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NR.sup.5)R.sup.6,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.NNR.sup.5R.sup.6),
C(.dbd.O)OR.sup.5, C(.dbd.O)NR.sup.5R.sup.6,
C(.dbd.O)NR.sup.5OR.sup.6, C(.dbd.O)NR.sup.5NR.sup.6R.sup.7,
C(.dbd.NR.sup.5)NR.sup.6R.sup.7, C(.dbd.NOR.sup.5)NR.sup.6R.sup.7
or R.sup.8, wherein,
[0014] R.sup.5, R.sup.6, and R.sup.7 independently is H, alkyl,
alkenyl, alkynyl, alkadienyl, cycloalkyl, cycloheteroalkyl, aryl or
heteroaryl, optionally substituted with one or more substituents
selected from the group consisting of halogen, CN, CNO,
C(halogen).sub.3, .dbd.O, OR.sup.8, OC(.dbd.O)R.sup.8,
OC(.dbd.O)OR.sup.8, OC(.dbd.O)NR.sup.8R.sup.9, SR.sup.8,
S(.dbd.O)R.sup.8, S(.dbd.O).sub.2R.sup.8,
S(.dbd.O).sub.2NR.sup.8R.sup.9, NO.sub.2, N.sub.3, NR.sup.8R.sup.9,
N.sup.+R.sup.8R.sup.9R.sup.10, NR.sup.5OR.sup.8,
NR.sup.5NR.sup.6R.sup.7, NR.sup.8C(.dbd.O)R.sup.9,
NR.sup.8C(.dbd.O)OR.sup.9, NR.sup.8C(.dbd.O)NR.sup.9R.sup.10, NC,
P(.dbd.O)(OR.sup.8)OR.sup.9, P.sup.+R.sup.5R.sup.6R.sup.7,
C(.dbd.O)R.sup.8, C(.dbd.NR.sup.8)R.sup.9,
C(.dbd.NOR.sup.8)R.sup.9, C(.dbd.NNR.sup.8R.sup.9),
C(.dbd.O)OR.sup.8, C(.dbd.O)NR.sup.8R.sup.9,
C(.dbd.O)NR.sup.8OR.sup.9 C(.dbd.NR.sup.5)NR.sup.6R.sup.7,
C(.dbd.NOR.sup.5)NR.sup.6R.sup.7 or
C(.dbd.O)NR.sup.8NR.sup.9R.sup.10, wherein R.sup.5 and R.sup.3 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring, wherein,
[0015] R.sup.8, R.sup.9, and R.sup.10 independently is H, alkyl,
alkenyl, alkynyl, alkadienyl, cycloalkyl, 20 cycloheteroalkyl, aryl
or heteroaryl and wherein R.sup.8 and R.sup.9 may together form a
3-8 membered heterocyclic ring or R.sup.8 and R.sup.10 may together
form a 3-8 membered heterocyclic ring or R.sup.9 and R.sup.10may
together form a 3-8 membered heterocyclic ring.
[0016] In the present description and claims, the direction of
connections between the various components of a building block
should be read left to right. For example an S--C-connecting group
--C(.dbd.O)--NH-- is connected to a Spacer through the carbon atom
on the left and to a Carrier through the nitrogen atom on the right
hand side.
[0017] The term "C.sub.3-C.sub.7 cycloheteroalkyl" as used herein
refers to a radical of totally saturated heterocycle like a cyclic
hydrocarbon containing one or more heteroatoms selected from
nitrogen, oxygen, phosphor, boron and sulphur independently in the
cycle such as pyrrolidine (1-pyrrolidine; 2-pyrrolidine;
3-pyrrolidine; 4-pyrrolidine; 5-pyrrolidine); pyrazolidine
(1-pyrazolidine; 2-pyrazolidine; 3-pyrazolidine; 4-pyrazolidine;
5-pyrazolidine); imidazolidine (1-imidazolidine; 2-imidazolidine;
3-imidazolidine; 4 imidazolidine; 5-imidazolidine); thiazolidine
(2-thiazolidine; 3-thiazolidine; 4-thiazolidine; 5-thiazolidine);
piperidine (1-piperidine; 2-piperidine; 3-piperidine; 4-piperidine;
5-piperidine; 6-piperidine); piperazine (1-piperazine;
2-piperazine; 3-piperazine; 4-piperazine; 5-piperazine;
6-piperazine); morpholine (2-morpholine; 3-morpholine;
4-morpholine; 5-morpholine; 6-morpholine); thiomorpholine
(2-thiomorpholine; 3-thiomorpholine; 4-thiomorpholine;
5-thiomorpholine; 6-thiomorpholine); 1,2-oxathiolane
(3-(1,2-oxathiolane); 4-(1,2-oxathiolane); 5-(1,2-oxathiolane);
1,3-dioxolane (2-(1,3-dioxolane); 4-(1,3-dioxolane);
5(1,3dioxolane); tetrahydropyrane; (2-tetrahydropyrane;
3-tetrahydropyrane; 4-tetrahydropyrane; 5-tetrahydropyrane;
6-tetrahydropyrane); hexahydropyridazine (1-(hexahydropyridazine);
2-(hexahydropyndazine); 3-(hexahydropyridazine);
4-(hexahydropyridazine); 5-(hexahydropyridazine);
6-(hexahydropyridazine)), [1,3,2]dioxaborolane,
[1,3,6,2]dioxazaborocane
[0018] The term "aryl" as used herein includes carbocyclic aromatic
ring systems of 5-7 carbon atoms. Aryl is also intended to include
the partially hydrogenated derivatives of the carbocyclic systems
as well as up to four fused fused aromatic- or partially
hydrogenated rings, each ring comprising 5-7 carbon atoms.
[0019] The term "heteroaryl" as used herein includes heterocyclic
unsaturated ring systems containing, in addition to 2-18 carbon
atoms, one or more heteroatoms selected from nitrogen, oxygen and
sulphur such as furyl, thienyl, pyrrolyl, heteroaryl is also
intended to include the partially hydrogenated derivatives of the
heterocyclic systems enumerated below.
[0020] The terms "aryl" and "heteroaryl" as used herein refers to
an aryl which can be optionally substituted or a heteroaryl which
can be optionally substituted and includes phenyl, biphenyl,
indenyl, naphthyl (1-naphthyl, 2-naphthyl), N-hydroxytetrazolyl,
N-hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl
(1-anthracenyl, 2-anthracenyl, 3-anthracenyl), thiophenyl
(2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl), indolyl,
oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl,
isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl
(2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl,
2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl
(1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol-4-yl,
1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl),
thiazolyl (2-thiazolyl, 4-thiazolyl, 5-thiazolyl), pyridyl
(2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl,
4-pyrimidinyl, 5-pyrimidinyl, 6pyrimidinyl), pyrazinyl, pyridazinyl
(3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl), quinolyl
(2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6 quinolyl,
7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl,
4-isoquinolyl, 5-isoquinolyl, 6-isoquinolyl, 7-isoquinolyl,
8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl,
3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl,
6-benzo[b]furanyl, 7-benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl
(2-(2,3-dihydro-benzo[b]f- uranyl),
3-(2,3-dihydro-benzo[b]furanyl), 4-(2,3-dihydro-benzo[b]furanyl),
5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro-benzo[b]furanyl),
7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl
(2-benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl,
5-benzo[b]thiophenyl, 6-benzo[b]thiophenyl, 7-benzo[b]thiophenyl),
2,3-dihydro-benzo[b]thiophenyl (2-(2,3-dihydro-benzo[b]thiophenyl),
3-(2,3-dihydro-benzo[b]thiophenyl),
4-(2,3-dihydro-benzo[b]thiophenyl),
5-(2,3-dihydro-benzo[b]thiophenyl),
6-(2,3-dihydro-benzo[b]thiophenyl),
7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl,
3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole
(1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl,
7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl,
4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl,
7-benzimidazolyl, 8-benzimidazolyI), benzoxazolyl (1-benzoxazolyl,
2-benzoxazolyl), benzothiazolyl (1-benzothiazolyl,
2-benzothiazolyl, 4-benzothiazolyl, 5-benzothiazolyl,
6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl,
2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine
(5H-dibenz[b,f]azepin-1-yl, 5H-dibenz[b,f]azepine-2- -yl,
5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl,
5H-dibenz[b,f]azepine-5-yl), 10,11-dihydro-5H-dibenz[b,f]azepine
(10,11-dihydro-5H-dibenz[b,f]azepine-1-yl,
10,11-dihydro-5H-dibenz[b,f]az- epine-2-yl,
10,11-dihydro-5H-dibenz[b,f]azepine-3-yl, 10,11
-dihydro-5H-dibenz[b,f]azepine-4-yl,
10,11-dihydro-5H-dibenz[b,f]azepine-- 5-yl).
[0021] The Functional Entity carries elements used to interact with
host molecules and optionally reactive elements allowing further
elaboration of an encoded molecule of a library. Interaction with
host molecules like enzymes, receptors and polymers is typically
mediated through van der waal's interactions, polar- and ionic
interactions and pi-stacking effects. Substituents mediating said
effects may be masked by methods known to an individual skilled in
the art (Greene, T. W.; Wuts, P. G. M. Protective Groups in Organic
Synthesis; 3rd ed.; John Wiley & Sons: New York, 1999.) to
avoid undesired interactions or reactions during the preparation of
the individual building blocks and during library synthesis.
Analogously, reactive elements may be masked by suitably selected
protection groups. It is appreciated by one skilled in the art that
by suitable protection, a functional entity may carry a wide range
of substitutents.
[0022] The Functional Entity Precursor is a masked. Functional
Entity that is incorporated into an encoded molecule. After
incorporation, reactive elements of the Functional Entity may be
revealed by un-masking allowing further synthetic operations.
Finally, elements mediating recognition of host molecules may be
un-masked.
[0023] In a certain aspect of the invention, Functional entity
precursor is --C(H)(R.sup.11)--R.sup.11' or functional entity
precursor is heteroaryl or aryl substituted with 0-3 R.sup.11, 0-3
R.sup.13 and 0-3 R.sup.15, wherein
[0024] R.sup.11 and R.sup.11' are independently H, or selected
among the group consisting of a C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.4-C.sub.8
alkadienyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cyclo-heteroalkyl, aryl, and heteroaryl, said group being
substituted with 0-3 R.sup.12, 0-3 R.sup.13 and 0-3 R.sup.15,
[0025] or R.sup.11 and R.sup.11'0 are C.sub.1-C.sub.3
alkylene-NR.sup.12.sub.2, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)R.sup.16, C.sub.1-C.sub.3
alkylene-NR.sup.12C(O)OR.sup.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12.sub.2, C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)R.s- up.16, C.sub.1-C.sub.2
alkylene-O--NR.sup.12C(O)OR.sup.16 substituted with 0-3
R.sup.15,
[0026] where R.sup.12 is H or selected independently among the
group consisting of C.sub.1C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl, heteroaryl, said group
being substituted with 0-3 R.sup.13 and 0-3 R.sup.15,
[0027] R.sup.13 is selected independently from --N.sub.3, --CNO,
--C(NOH)NH.sub.2, --NHOH, --NHNHR.sup.17, --C(O)R.sup.17,
--SnR.sup.17.sub.3, --B(OR.sup.17).sub.2, --P(O)(OR.sup.17).sub.2
or the group consisting of C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.4-C.sub.8 alkadienyl said group being substituted
with 0-2 R.sup.14,
[0028] where R.sup.14 is independently selected from --NO.sub.2,
--C(O)OR.sup.17, --COR.sup.17, --CN, --OSiR.sup.17.sub.3,
--OR.sup.17 and --NR.sup.17.sub.2;
[0029] R.sup.15 is .dbd.O, --F, --Cl, --Br, --I, --CN, --NO.sub.2,
--OR.sup.17, --NR.sup.17.sub.2, --NR.sup.17--C(O)R.sup.16,
--NR.sup.17--C(O)OR.sup.16, --SR.sup.17, --S(O)R.sup.17,
--S(O).sub.2R.sup.17, --COOR.sup.17, --C(O)NR.sup.17.sub.2 and
--S(O).sub.2NR.sup.17.sub.2,
[0030] R.sup.16 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.7 cycloalkyl, aryl
or C.sub.1-C.sub.6 alkylene-aryl substituted with 0-3 substituents
independently selected from --F, --Cl, --NO.sub.2, --R.sup.2,
--R.sup.2, --SiR.sup.2.sub.3;
[0031] R.sup.17 is selected independently from H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, aryl, C.sub.1-C.sub.6
alkylene-aryl, 1
[0032] G is H or C.sub.1-C.sub.6 alkyl and n is 1,2,3 or 4.
[0033] The function of the carrier is to ensure the transferability
of the functional entity precursor. To adjust the transferability a
skilled chemist can design suitable substitutions of the carrier by
evaluation of initial attempts. The transferability may be adjusted
in response to the chemical composition of the functional entity
precursor, to the nature of the complementing element, to the
conditions under which the transfer and recognition is performed,
etc.
[0034] In a preferred embodiment, the carrier is selected from the
group consisting of arylene, heteroarylene or --(CF.sub.2).sub.m--
substituted with 0-3 R.sup.1 wherein m is an integer between 1 and
10, and C--F-connecting group is --SO.sub.2--O--. Due to the high
reactivity of such compounds a broad range of recipient reactive
groups may be employed in the construction of carbon-carbon bonds
or carbon-hetero atom bonds.
[0035] In another preferred embodiment of the invention, the
carrier is --(CF.sub.2).sub.m-- wherein m is an integer between 1
and 10, the C--F-connecting group is --SO.sub.2--O--; and the
functional entity precursor is aryl or heteroaryl substituted with
0-3 R.sup.11, 0-3 R.sup.13 and 0-3 R.sup.15.
[0036] The C--F-connecting group determines in concert with the
carrier the transferability of the functional entity precursor. In
a preferred embodiment, the C--F-connecting group is
--S.sup.+(R.sup.11)--,
[0037] In another preferred embodiment, the C--F-connecting group
is chosen from the group consisting of --SO.sub.2--O--, and
--S.sup.+(R.sup.17)--; wherein R.sup.17 is selected independently
from H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, aryl,
C.sub.1-C.sub.6 alkylene-aryl.
[0038] In the presence of a catalyst comprising transition metals
such as Pd, Ni or Cu, an aromatic moiety may be transferred from
the C--F-connecting group to a recipient reactive group. Further,
the transfer may be initiated by adding the catalyst, independently
of the annealing of encoding - and complementing elements.
[0039] The S--C-connecting group provide a means for connecting the
Spacer and the Carrier. As such it is primarily of synthetic
convenience and does not influence the function of a building
block.
[0040] The spacer serves to distance the functional entity
precursor to be transferred from the bulky complementing element.
Thus, when present, the identity of the spacer is not crucial for
the function of the building block. It may be desired to have a
spacer which can be cleaved by light. In this case, the spacer is
provided with e.g. the group 2
[0041] In the event an increased hydrophilicity is desired the
spacer may be provided with a polyethylene glycol part of the
general formula: 3
[0042] In a preferred embodiment, the complementing element serves
the function of transferring genetic information e.g. by
recognising a coding element. The recognition implies that the two
parts are capable of interacting in order to assemble a
complementing element--coding element complex. In the
biotechnological field a variety of interacting molecular parts are
known which can be used according to the invention. Examples
include, but are not restricted to protein-protein interactions,
protein-polysaccharide interactions, RNA-protein interactions,
DNA-DNA interactions, DNA-RNA interactions, RNA-RNA interactions,
biotin-streptavidin interactions, enzyme-ligand interactions,
antibody-ligand interaction, protein-ligand interaction, etc.
[0043] The interaction between the complementing element and coding
element may result in a strong or a weak bonding. If a covalent
bond is formed between the parties of the affinity pair the binding
between the parts can be regarded as strong, whereas the
establishment of hydrogen bondings, interactions between
hydrophobic domains, and metal chelation in general results in
weaker bonding. In general relatively weak bonding is preferred. In
a preferred aspect of the invention, the complementing element is
capable of reversible interacting with the coding element so as to
provide for an attachment or detachment of the parts in accordance
with the changing conditions of the media.
[0044] In a preferred aspect of the invention, the interaction is
based on nucleotides, i.e. the complementing element is a nucleic
acid. Preferably, the complementing element is a sequence of
nucleotides and the coding element is a sequence of nucleotides
capable of hybridising to the complementing element. The sequence
of nucleotides carries a series of nucleobases on a backbone. The
nucleobases may be any chemical entity able to be specifically
recognized by a complementing entity. The nucleobases are usually
selected from the natural nucleobases (adenine, guanine, uracil,
thymine, and cytosine) but also the other nucleobases obeying the
Watson-Crick hydrogen-bonding rules may be used, such as the
synthetic nucleobases disclosed in U.S. Pat. No. 6,037,120.
Examples of natural and non-natural nucleobases able to perform a
specific pairing are shown in FIG. 2. The backbone of the sequence
of nucleotides may be any backbone able to aggregate the
nucleobases is a sequence. Examples of backbones are shown in FIG.
4. In some aspects of the invention the addition of non-specific
nucleobases to the complementing element is advantegeous, FIG.
3
[0045] The coding element can be an oligonucleotide having
nucleobases which complements and is specifically recognised by the
complementing element, i.e. in the event the complementing element
contains cytosine, the coding element part contains guanine and
visa versa, and in the event the complementing element contains
thymine or uracil the coding element contains adenine.
[0046] The complementing element may be a single nucleobase. In the
generation of a library, this will allow for the incorporation of
four different functional entities into the template-directed
molecule. However, to obtain a higher diversity a complementing
element preferably comprises at least two and more preferred at
least three nucleotides. Theoretically, this will provide for 42
and 43, respectively, different functional entities uniquely
identified by the complementing element. The complementing element
will usually not comprise more than 100 nucleotides. It is
preferred to have complementing elements with a sequence of 3 to 30
nucleotides.
[0047] The building blocks of the present invention can be used in
a method for transferring a functional entity precursor to a
recipient reactive group, said method comprising the steps of
[0048] providing one or more building blocks as described above
and
[0049] contacting the one or more building blocks with a
corresponding encoding element associated with a recipient reactive
group under conditions which allow for a recognition between the
one or more complementing elements and the encoding elements, said
contacting being performed prior to, simultaneously with, or
subsequent to a transfer of the functional entity precursor to the
recipient reactive group.
[0050] The encoding element may comprise one, two, three or more
codons, i.e. sequences that may be specifically recognised by a
complementing element. Each of the codons may be separated by a
suitable spacer group. Preferably, all or at least a majority of
the codons of the template are arranged in sequence and each of the
codons are separated from a neighbouring codon by a spacer group.
Generally, it is preferred to have more than two codons on the
template to allow for the synthesis of more complex encoded
molecules. In a preferred aspect of the invention the number of
codons of the encoding element is 2 to 100. Still more preferred
are encoding elements comprising 3 to 10 codons. In another aspect,
a codon comprises 1 to 50 nucleotides and the complementing element
comprises a sequence of nucleotides complementary to one or more of
the encoding sequences.
[0051] The recipient reactive group may be associated with the
encoding element in any appropriate way. Thus, the reactive group
may be associated covalently or noncovalently to the encoding
element. In one embodiment the recipient reactive group is linked
covalently to the encoding element through a suitable linker which
may be separately cleavable to release the reaction product. In
another embodiment, the reactive group is coupled to a
complementing element, which is capable of recognising a sequence
of nucleotides on the encoding element, whereby the recipient
reactive group becomes attached to the encoding element by
hybridisation. Also, the recipient reactive group may be part of a
chemical scaffold, i.e. a chemical entity having one or more
reactive groups available for receiving a functional entity
precursor from a building block.
[0052] The recipient reactive group may be any group able to
participate in cleaving the bond between the carrier and the
functional entity precursor to release the functional entity
precursor. Typically, the recipient reactive group is a
nucleophilic atom such as S, N, 0, C or P. Scheme 1a shows the
transfer of an alkyl group and scheme 1b shows the transfer of an
vinyl group. 4 5
[0053] Alternatively, the recipient reactive group is a
organometallic compound as shown in scheme 2. 6
[0054] According to a preferred aspect of the invention the
building blocks are used for the formation of a library of
compounds. The complementing element of the building block is used
to identify the functional entity. Due to the enhanced proximity
between reactive groups when the complementing entity and the
encoding element are contacted, the functional entity precursor
together with the identity programmed in the complementing element
is transferred to the encoding element associated with recipient
reactive group. Thus, it is preferred that the sequence of the
complementing element is unique in the sense that the same sequence
is not used for another functional entity. The unique
identification of the functional entity enable the possibility of
decoding the encoding element in order to determine the synthetic
history of the molecule formed. In the event two or more functional
entities have been transferred to a scaffold, not only the identity
of the transferred functional entities can be determined. Also the
sequence of reaction and the type of reaction involved can be
determined by decoding the encoding element. Thus, according to a
preferred embodiment of the invention, each different member of a
library comprises a complementing element having a unique sequence
of nucleotides, which identifies the functional entity.
BRIEF DESCRIPTION OF THE DRAWINGS
[0055] FIG. 1. Two setups for Functional Entity Transfer
[0056] FIG. 2. Examples of specific base pairing
[0057] FIG. 3. Example of non-specific base-pairing
[0058] FIG. 4. Backbone examples
[0059] FIG. 5 Three examples of building blocks
DETAILED DESCRIPTION OF THE INVENTION
[0060] A building block of the present invention is characterized
by its ability to transfer its functional entity precursor to a
recipient reactive group. This is done by forming a new covalent
bond between the recipient reactive group and cleaving the bond
between the carrier moiety and the functional entity precursor of
the building block.
[0061] Two setups for generalized functional entity precursor
transfer from a building block are depicted in FIG. 1. In the first
example, one complementing element of a building block recognizes a
coding element carrying another functional entity precursor, hence
bringing the functional entities in close proximity. This results
in a reaction between functional entity precursor 1 and 2 forming a
covalent bond between these concurrent with the cleavage of the
bond between functional entity precursor 2 and its linker. In the
second example, a template brings together two building blocks
resulting in functional entity precursor transfer from one building
block to the other.
[0062] FIG. 5 illustrates three specific compounds according to the
invention. For illustrative purposes the individual features used
in the claims are indicated. The upper compound is an example of a
building block wherein the linker is backbone attached at the
3'-position. The first part of the linker, i.e. the spacer, is an
aliphatic chain ending in a nitrogen atom. The nitrogen atom
bridges to the S--C-connecting group, which is an N-acylated
arylmethyleamine. The carrier attached to the left hand side
carbonyl group of the S--C-connecting group is a benzene ring
holding the C--F Connecting group in the para position. The C--F
Connecting group is a positively charged sulfur atom which is
attached to the Functional Entity Precursor, in this case a benzyl
group. When the building block is presented to a nucleophilic
recipient reactive group, such an amine or a thiol, Functional
Entity Precursor is transferred to benzylate the recipient reactive
group.
[0063] The middle compound illustrates a 5'0 attachment of a
linker. The linker is linked through a phosphate group and extends
into a three membered aliphatic chain. Through another phosphate
group and a PEG linker the complementing element is linked via an
amide bond to the Carrier. When the building block is presented to
a nucleophile the Functional Entity Precursor is transferred
resulting in an alkylation of the nucleophile.
[0064] The lower compound illustrates a nucleobase attachment of
the linker. The linker attaches to the 5 position of a pyrimidine
type nucleobase and extents through an .alpha.-.beta. unsaturated
N-methylated amide to the S--C-connecting group, which is a 4-amino
methyl benzoic acid derivative. The functional entity precursor can
be transferred to a nucleophilic recipient reactive group e.g. an
amine or a thiol forming an allylic amine or thiol.
[0065] According to the invention, the functional entity precursor
is of the formula --C(H)(R.sup.3)--R.sup.4 or functional entity
precursor is heteroaryl or aryl optionally substituted with one or
more substituents belonging to the group comprising R.sup.3 and
R.sup.4. In a further preferred embodiment,
[0066] R.sup.3 and R.sup.4 independently is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.4-C.sub.8 alkadienyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl or heteroaryl, optionally
substituted with one or more substituents selected from the group
consisting of
[0067] SnR.sup.5R.sup.6, R.sup.7, Sn(OR.sup.5)R.sup.6R.sup.7,
Sn(OR.sup.5)(OR.sup.6)R.sup.7, BR.sup.5R.sup.6, B(OR.sup.5)R.sup.6,
B(OR.sup.5)(OR.sup.6), halogen, CN, CNO, C(halogen).sub.3, .dbd.O,
OR.sup.5, OC(.dbd.O)R.sup.5, OC(.dbd.O)OR.sup.5,
OC(.dbd.O)NR.sup.5R.sup.- 6, SR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
N.sub.3, NR.sup.5R.sup.6, N.sup.+R.sup.5R.sup.6R.sup.7,
NR.sup.5OR.sup.6, NR.sup.5NR.sup.6R.sup.7,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, NC, P(.dbd.O)(OR.sup.5)OR.sup.6,
P.sup.+R.sup.5R.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NR.sup.5)R.sup.6, C(.dbd.NOR.sup.5)R.sup.6,
C(.dbd.NNR.sup.5R.sup.6), C(.dbd.O)OR, C(.dbd.O)NR.sup.5R.sup.6,
C(.dbd.O)NR.sup.5OR.sub.6, C(.dbd.O)NR.sup.5NR.sup.6R.sup.7,
C(.dbd.NR.sup.5)NR.sup.6R.sup.7, C(.dbd.NOR.sup.5)NR.sup.6R.sup.7
or R.sup.8, wherein,
[0068] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.4-C.sub.8 alkadienyl, C.sub.3-C.sub.7 cycloalkyl,
C.sub.3-C.sub.7 cycloheteroalkyl, aryl or heteroaryl and wherein
R.sup.5 and R.sup.6 may together form a 3-8 membered heterocyclic
ring or R.sup.5 and R.sup.7 may together form a 3-8 membered
heterocyclic ring or R.sup.6 and R.sup.7 may together form a 3-8
membered heterocyclic ring,
[0069] in another prefered embodiment,
[0070] R.sup.3 and R.sup.4 independently is H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl, optionally substituted with
one or more substituents selected from the group consisting of
halogen, CN, C(halogen).sub.3, .dbd.O, OR.sup.5, OC(.dbd.O)R.sup.5,
OC(.dbd.O)OR.sup.5, OC(.dbd.O)NR.sup.5R.sup.- 6, SR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5OR.sup.6, NR5NR.sup.6R.sup.7, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
P(.dbd.O)(OR5)OR.sup.6, C(.dbd.O)R.sup.5, C(.dbd.NR.sup.5)R.sup.6,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.NNR.sup.5R.sup.6),
C(.dbd.O)OR.sup.5, C(.dbd.O)NR.sup.5R.sup.6,
C(.dbd.O)NR.sup.5OR.sup.6, C(.dbd.O)NR.sup.5NR.sup.6R.sup.7,
C(.dbd.NR.sup.5)NR.sup.6R.sup.7, C(.dbd.NOR.sup.5)NR.sup.6R.sup.7
or R.sup.8, wherein,
[0071] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0072] in still another prefered embodiment,
[0073] R.sup.3 and R.sup.4 independently is H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl, optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, OC(.dbd.O)R.sup.5,
OC(.dbd.O)OR.sup.5, OC(.dbd.O)NR.sup.5R.sup.6, SR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5R.sup.6, NR.sup.5NR.sup.6R.sup.7, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
P(.dbd.O)(OR.sup.5)OR.sup.6, C(.dbd.O)R.sup.6,
C(.dbd.NR.sup.5)R.sup.6, C(.dbd.NOR.sup.5)R.sup.6,
C(.dbd.NNR.sup.5R.sup.6), C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6,
C(.dbd.O)NR.sup.5NR.sup.6R.sup.7, C(.dbd.NR.sup.5)NR.sup.6R.sup.7,
C(.dbd.NOR.sup.5)NR.sup.6R.sup.7 or R.sup.8, wherein,
[0074] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0075] in still another prefered embodiment,
[0076] R.sup.3 and R.sup.4 independently is H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl, optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0077] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0078] in still another prefered embodiment,
[0079] R.sup.3 and R.sup.4 independently is H, methyl, ethyl,
propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl,
phenyl, naphtyl, thienyl, furyl, pyridyl, quinolinyl or
isoquinolinyl optionally substituted with one or more substituents
selected from the group consisting of F, Cl, CN, CF.sub.3, .dbd.O,
OR.sup.5, S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0080] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0081] in still another prefered embodiment,
[0082] R.sup.3 and R.sup.4 independently is H, methyl, ethyl,
propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0083] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.6 and R5 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0084] in still another prefered embodiment,
[0085] R.sup.3 and R.sup.4 independently is H, aziridinyl,
azetidinyl, pyrrolidinyl, piperidinyl or morpholinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0086] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 38 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0087] in still another prefered embodiment,
[0088] R.sup.3 and R.sup.4 independently is H, phenyl, naphtyl,
thienyl, furyl, pyridyl, quinolinyl or isoquinolinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.5,
wherein,
[0089] R.sup.5, R.sup.6, R.sup.7 and R.sup.3 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0090] in still another prefered embodiment,
[0091] R.sup.3 and R.sup.4 independently is H, phenyl or naphtyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0092] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0093] in still another prefered embodiment,
[0094] R.sup.3 and R.sup.4 independently is H, thienyl, furyl,
pyridyl, quinolinyl or isoquinolinyl optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, R.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0095] R.sup.5, R.sup.6, R.sup.7 and R.sup.5 independently is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl and wherein R.sup.5 and
R.sup.6 may together form a 3-8 membered heterocyclic ring or
R.sup.5 and R.sup.7 may together form a 3-8 membered heterocyclic
ring or R.sup.6 and R.sup.7 may together form a 3-8 membered
heterocyclic ring,
[0096] in still another prefered embodiment,
[0097] R.sup.3 and R.sup.4 independently is H, methyl, ethyl,
propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0098] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl
or isoquinolinyl and wherein R.sup.5 and R.sup.6 may together form
a 3-8 membered heterocyclic ring or R.sup.5 and R.sup.7 may
together form a 3-8 membered heterocyclic ring or R.sup.6 and
R.sup.7 may together form a 3-8 membered heterocyclic ring,
[0099] in still another prefered embodiment,
[0100] R.sup.3 and R.sup.4 independently is H, aziridinyl,
azetidinyl, pyrrolidinyl, piperidinyl or morpholinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0101] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl
or isoquinolinyl and wherein R.sup.5 and R.sup.6 may together form
a 3-8 membered heterocyclic ring or R.sup.5 and R.sup.7 may
together form a 3-8 membered heterocyclic ring or R.sup.6 and
R.sup.7 may together form a 3-8 membered heterocyclic ring,
[0102] in still another prefered embodiment,
[0103] R.sup.3 and R.sup.4 independently is H, phenyl, naphtyl,
thienyl, furyl, pyridyl, quinolinyl or isoquinolinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5s, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5, C(.dbd.O)NRBR.sup.6,
C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8, wherein,
[0104] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl
or isoquinolinyl and wherein R.sup.5 and R.sup.6 may together form
a 3-8 membered heterocyclic ring or R.sup.5 and R.sup.7 may
together form a 3-8 membered heterocyclic ring or R.sup.6 and
R.sup.7 may together form a 3-8 membered heterocyclic ring,
[0105] in still another prefered embodiment,
[0106] R.sup.3 and R.sup.4 independently is H, phenyl or naphtyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0107] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl
or isoquinolinyl and wherein R.sup.5 and R.sup.6 may together form
a 3-8 membered heterocyclic ring or R.sup.5 and R.sup.7 may
together form a 3-8 membered heterocyclic ring or R.sup.6 and
R.sup.7 may together form a 3-8 membered heterocyclic ring,
[0108] in still another prefered embodiment,
[0109] R.sup.3 and R.sup.4 independently is H, thienyl, furyl,
pyridyl, quinolinyl or isoquinolinyl optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0110] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl
or isoquinolinyl and wherein R.sup.5 and R.sup.6 may together form
a 38 membered heterocyclic ring or R.sup.5 and R.sup.7 may together
form a 3-8 membered heterocyclic ring or R.sup.6 and R.sup.7 may
together form a 3-8 membered heterocyclic ring,
[0111] in still another prefered embodiment,
[0112] R.sup.3 and R.sup.4 independently is H, methyl, ethyl,
propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0113] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl or butyl and wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0114] in still another prefered embodiment,
[0115] R.sup.3 and R.sup.4 independently is H, aziridinyl,
azetidinyl, pyrrolidinyl, piperidinyl or morpholinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NRR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0116] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl or butyl and wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0117] in still another prefered embodiment,
[0118] R.sup.3 and R.sup.4 independently is H, phenyl, naphtyl,
thienyl, furyl, pyridyl, quinolinyl or isoquinolinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0119] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl or butyl and wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0120] in still another prefered embodiment,
[0121] R.sup.3 and R.sup.4 independently is H, phenyl or naphtyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3,.dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0122] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl or butyl and wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0123] in still another prefered embodiment,
[0124] R.sup.3 and R.sup.4 independently is H, thienyl, furyl,
pyridyl, quinolinyl or isoquinolinyl optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.6C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0125] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
methyl, ethyl, propyl or butyl and wherein R.sup.5 and R.sup.6 may
together form a 3-8 membered heterocyclic ring or R.sup.5 and
R.sup.7 may together form a 3-8 membered heterocyclic ring or
R.sup.6 and R.sup.7 may together form a 3-8 membered heterocyclic
ring,
[0126] in still another prefered embodiment,
[0127] R.sup.3 and R.sup.4 independently is methyl, ethyl, propyl,
butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.6R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.6)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0128] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,
[0129] in still another prefered embodiment,
[0130] R.sup.3 and R.sup.4 independently is aziridinyl, azetidinyl,
pyrrolidinyl, piperidinyl or morpholinyl optionally substituted
with one or more substituents selected from the group consisting of
F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein, R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,
[0131] in still another prefered embodiment,
[0132] R.sup.3 and R.sup.4 independently is phenyl, naphtyl,
thienyl, furyl, pyridyl, quinolinyl or isoquinolinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0133] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,
[0134] in still another prefered embodiment,
[0135] R.sup.3 and R.sup.4 independently is phenyl or naphtyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0136] R.sup.5, R.sup.8, R.sup.7 and R.sup.8 independently is H,
cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,
[0137] in still another prefered embodiment,
[0138] R.sup.3 and R.sup.4 independently is thienyl, furyl,
pyridyl, quinolinyl or isoquinolinyl optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0139] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,
[0140] in still another prefered embodiment,
[0141] R.sup.3 and R.sup.4 independently is methyl, ethyl, propyl,
butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0142] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl or
isoquinolinyl,
[0143] in still another prefered embodiment,
[0144] R.sup.3 and R.sup.4 independently is aziridinyl, azetidinyl,
pyrrolidinyl, piperidinyl or morpholinyl optionally substituted
with one or more substituents selected from the group consisting of
F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.6,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0145] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl or
isoquinolinyl,
[0146] in still another prefered embodiment,
[0147] R.sup.3 and R.sup.4 independently is phenyl, naphtyl,
thienyl, furyl, pyridyl, quinolinyl or isoquinolinyl optionally
substituted with one or more substituents selected from the group
consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0148] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl or
isoquinolinyl,
[0149] in still another prefered embodiment,
[0150] R.sup.3 and R.sup.4 independently is phenyl or naphtyl
optionally substituted with one or more substituents selected from
the group consisting of F, Cl, CN, CF.sub.3, .dbd.O, OR.sup.5,
S(.dbd.O)R.sup.5, S(.dbd.O).sub.2R.sup.5,
S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2, NR.sup.5R.sup.6,
NR.sup.5C(.dbd.O)R.sup.6, NR.sup.5C(.dbd.O)OR.sup.6,
NR.sup.5C(.dbd.O)NR.sup.6R.sup.7, C(.dbd.O)R.sup.5,
C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0151] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl or
isoquinolinyl,
[0152] in still another prefered embodiment,
[0153] R.sup.3 and R.sup.4 independently is thienyl, furyl,
pyridyl, quinolinyl or isoquinolinyl optionally substituted with
one or more substituents selected from the group consisting of F,
Cl, CN, CF.sub.3, .dbd.O, OR.sup.5, S(.dbd.O)R.sup.5,
S(.dbd.O).sub.2R.sup.5, S(.dbd.O).sub.2NR.sup.5R.sup.6, NO.sub.2,
NR.sup.5R.sup.6, NR.sup.5C(.dbd.O)R.sup.5,
NR.sup.5C(.dbd.O)OR.sup.6, NR.sup.5C(.dbd.O)NR.sup.6R.sup.7,
C(.dbd.O)R.sup.5, C(.dbd.NOR.sup.5)R.sup.6, C(.dbd.O)OR.sup.5,
C(.dbd.O)NR.sup.5R.sup.6, C(.dbd.O)NR.sup.5OR.sup.6 or R.sup.8,
wherein,
[0154] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 independently is H,
phenyl, naphthyl, thienyl, furyl, pyridinyl, quinolinyl or
isoquinolinyl,
[0155] in still another prefered embodiment,
[0156] R.sup.3 and R.sup.4 independently is H, C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.7 cycloalkyl, C.sub.3-C.sub.7
cycloheteroalkyl, aryl or heteroaryl
[0157] in still another prefered embodiment,
[0158] R.sup.3 and R.sup.4 independently is H,
[0159] in still another prefered embodiment,
[0160] R.sup.3 and R.sup.4 independently is C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.7 cycloalkyl or C.sub.3-C.sub.7 cycloheteroalkyl,
[0161] in still another prefered embodiment,
[0162] R.sup.3 and R.sup.4 independently is methyl, ethyl, propyl
or butyl
[0163] in still another prefered embodiment
[0164] R.sup.3 and R.sup.4 independently is cyclopropyl,
cyclobutyl, cyclopentyl or cyclohexyl
[0165] in still another prefered embodiment
[0166] R.sup.3 and R.sup.4 independently is aziridinyl,
pyrrolidinyl, piperidinyl or morpholinyl
[0167] in still another prefered embodiment,
[0168] R.sup.3 and R.sup.4 independently is aryl or heteroaryl
[0169] in still another prefered embodiment,
[0170] R.sup.3 and R.sup.4 independently is phenyl or naphthyl
[0171] in still another prefered embodiment,
[0172] R.sup.3 and R.sup.4 independently is thienyl, furyl,
pyridyl, quinolinyl or isoquinolyl
[0173] Experimental Section
[0174] General Procedure 1: Preparation of Carrier-Functional
Entity Reagents: 7
[0175] The 4-halobenzoic acid (25 mmol) is added to a ice cooled
solution of chloro sulfonic acid (140 mmol). The mixture is slowly
heated to reflux and left at reflux for 2-3 hours. The mixture is
added to 100 mL ice and the precipitate collected by filtration.
The filtrate is washed with water (2.times.50 mL) and the dried in
vacuo affording the corresponding sulfonoyl chloride in 60-80%
yield. The 3-chlorosulfonyl-4-halo-benzoic acid derivate (5 mmol)
is dissolved in EtOH (5 mL) and added to a ice cooled mixture of
NaOEt (10 mL, 2M). The mixture is stirred o/n at rt. Acetic acid
(40 mmol) is added and the mixture is evaporated in vacuo. Water
(10 mL) is added and pH adjusted to pH=2 (using 1M HCl). The
product is extracted with DCM (2.times.25 mL), dried over
Na.sub.2SO.sub.4 and evaporated in vacuo affording the desired
products.
EXAMPLE 1
General Procedure (1)
[0176] 3-Ethoxysulfonyl4-fluorobenzoic acid 8
[0177] .sup.1H-NMR (DMSO-d.sub.6): .delta. 8.49 (d, 1H), 7.85 (dd,
1H), 7.5 (d, 1H), 4.32 (q, 2H), 1.32 (t, 3H)
EXAMPLE 2
General Procedure (1)
[0178] 4-chloro-3-Ethoxysulfonylbenzoic acid 9
[0179] .sup.1H-NMR (DMSO-d.sub.6): .delta. 8.49 (d, 1H), 7.85 (dd,
1H), 7.5 (d, 1H), 4.32 (q, 2H), 1.32 (t, 3H)
EXAMPLE 3
[0180] 10
[0181] 4-Methylsulfanyl benzoic acid (0.5 g, 2.97 mmol,
commercially available from Aldrich, cat no. 145521) was added to
methyl p-toluene solfunate (0.61 g, 3.27 mmol). The mixture was
heated to 140.degree. C. for 1 hour in a sealed vessel. After
cooling to rt the mixture was trituated with diethyl ether.
Filtration and drying in vacuo yielded 844 mg (80%) of the desired
product (>95% pure by .sup.1H nmr).
[0182] .sup.1H nmr (DMSO-d6): 8.20-8.10 (m, 4H), 7.45 (d, 2H), 7.08
(d, 2H), 3.29 (s, 6H), 2.30 (s, 3H).
[0183] General Procedure 2: Solid Phase Preparation of
Carrier-Functional Entity Reagents for Alkylation Building Blocks:
11
[0184] Ps=Polystyrene resin. Alternatively other acid labile
linkers may be employed.
[0185] Step 1:
[0186] A polystyrene resin with a wang linker
(4-hydroxymethylphenol linker)(50 mg.about.50 umol), a
bi-functional carrier (200 umol, 4 equiv) in a solvent such as THF,
DCM, DCE, DMF, NMP or a mixture thereof (500 uL) and a base such as
TEA, DIEA, pyridine (400 umol, 8 equiv), optionally in the presence
of DMAP (100 umol), are allowed to react at temperatures between
-20.degree. C. and 60.degree. C., preferably between 0.degree. C.
and 25.degree. C., for 1-24 h, preferably 14 h. The resin is washed
with the solvent composition used during the reaction (5.times.1
mL) and used in the following step.
[0187] Step 2:
[0188] A functional entity precursor carrying a hydroxy group in
the position of the intended attachment to the C--F-connecting
group (200 umol, 4 equiv) in a solvent such as THF, DCM, DCE, DMF,
NMP or a mixture thereof (500 uL) and a base such as TEA, DIEA,
pyridine (400 umol, 8 equiv), optionally in the presence of DMAP,
are added to the resin bound carrier isolated in step 1 and allowed
to react at temperatures between 0.degree. C. and 100.degree. C.,
preferably between 25.degree. C. and 80.degree. C., for 248 h,
preferably. 4-16 h. The resin is washed with the solvent
composition used during the reaction (5.times.1 mL).
[0189] Step 3:
[0190] The desired Carrier-Functional entity reagent is cleaved
from the resin obtained in step 2 by treatment with an acid like
TFA, HF or HCl in a solvent such as THF, DCM, DCE or a mixture
thereof (1 mL) at temperatures between -20.degree. C. and
60.degree. C., preferably between 0.degree. C. and 25.degree. C.,
for 14 h, preferably 1-2 h. Upon filtration, the resin is washed
with the solvent composition used during cleavage (2.times.1 mL)
and the combined filtrates are evaporated in vacuo. The isolated
product may be purified by chromatography.
Assembly of Building Blocks
[0191] The Carrier-Functional entity reagent may be bound to the
Spacer by several different reactions as illustrated below.
Formation of an Amide Bond Between a Carboxylic Acid of the Carrier
and an Amine Group of a Spacer
[0192] 12
[0193] General Procedure 3: Preparation of Building Blocks by
Loading a Carrier-Functional Entity Reagent onto a Nucleotide
Derivative Comprising an Amino Group: 13
[0194] 15 uL of a 150 mM building block solution of
FE.sup.1-Carrier-COOH is mixed with 15 .mu.L of a 150 mM solution
of EDC and 15 .mu.L of a 150 mM solution of N-hydroxy-succinimide
(NHS) using solvents like DMF, DMSO, water, acetonitril, THF, DCM,
methanol, ethanol or a mixture thereof. The mixture is left for 15
min at 25.degree. C. 45 .mu.L of an aminooligo (10 nmol) in 100 mM
buffer at a pH between 5 and 10, preferably 6.0-7.5 is added and
the reaction mixture is left for 2 hours at 25.degree. C. Excess
building block and organic by-products were removed by extraction
with EtOAc (400 .mu.L). Remaining EtOAc is evaporated in vacuo
using a speedvac. The building block is purified following elution
through a BiORad micro-spin chromatography column, and analyzed by
electron spray mass spectrometry (ES-MS).
EXAMPLE 4
General Procedure ( )
[0195] 14
[0196] Where Oligo is 5'0 XCG ATG GAT GCT CCA GGT CGC 3', X=5'
amino C.sub.6 (Glen catalogue# 10-1906-90), Expected molecular
weight: 6313.22 MS (calc.)=6543,43; MS (found)=6513,68* * Observed
molecular weight of the cleaved sulfonic ester: 6513.68 Expected
molecular weight of the cleaved ester. 6514.37 The quantitative
loss of the ethyl group Is probably due to the presence of
pipeddine during the recording of the LCMS data.
[0197] General Procedure 4: Loading of a Carrier Coupled Functional
Entity onto an Amino Ontgo:
[0198] 25 .mu.l 100 mM carrier coupled functional entity dissolved
in DMF (dimethyl formamide) was mixed with 25 .mu.l 100 mM EDC
(1-ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride) in
DMF for 30 minutes at 25.degree. C. The mixture was added to 50
.mu.l amino oligo in H.sub.2O with 100 mM HEPES
(2-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-eth- anesulfonic acid) pH
7.5 and the reaction was allowed to proceed for 20 minutes at
25.degree. C. Unreacted carrier coupled functional entity was
removed by extraction with 500 .mu.l EtOAc (ethyl acetate), and the
oligo was purified by gel filtration through a microspin column
equilibrated with 100 mM MES (2-(N-morpholino) ethanesulfonic acid)
pH 6.0.
[0199] Oligonucleotide used:
[0200] Oligo A: 5'-YACGATGGATGCTCCAGGTCGC
[0201] Y=Amino modifier C6 (Glen#10-1906)
EXAMPLE 5
General Procedure 4
[0202] Carrier--Functional Entity:
(4-Carboxy-phenyl)-dimethyl-sulfonium 15
[0203] Mass: 6789.21 (observed using ES-MS), 6790.65
(calculated)
[0204] General Procedure 5: Preparation of Arylation Building
Blocks: 16
[0205] Funtional Entity-OH is a phenol, n is an integer between 3
and 6.
[0206] Step 1
[0207] To a solution of the bis-sulfonylchloride (Ward, R. B.; J.
Org. Chem.; 30; 1965; 3009-3011; Qiu, Weiming; Burton, Donald J.;
J. Fluorine Chem.; 60; 1; 1993; 93100)(3 umol) in DMF, DMSO,
acetonitril, THF or a mixture thereof (150 uL) is a phenolic
functional entity in excess (1.05-1.8 mmol) in DMF, DMSO,
acetonitril, THF or a mixture thereof (150 uL) added slowly at
temperatures between -20.degree. C. and 100.degree. C. preferably
at 0-50.degree. C. in the presence of a base such as TEA, DIEA,
pyridine, Na-HCO.sub.3 or K.sub.2CO.sub.3.
[0208] Step2
[0209] The reaction mixture from step 1 is added to a solution of
an aminooligo (10 nmol) in 100 mM buffer at a pH between 5 and 10,
preferably 6.0-7.5 optionally in the presence of NHS. The reaction
mixture is left for 2 hours at 25.degree. C. Excess building block
and organic by-products were removed by extraction with EtOAc (400
.mu.L). Remaining EtOAc is evaporated in vacuo using a speedvac.
The building aminooligo is purified following elution through a
BiORad micro-spin chromatography column, and analyzed by electron
spray mass spectrometry (ES-MS).
Use of Building Blocks
[0210] General Procedure 6: Alkylation of Oligonucleotide
Derivatives Containing a Nucleophilic Recipient Group Using a
Building Block of the Invention: 17
[0211] An oligonucleotide building block carrying functional entity
FE.sup.1 is combined at 2 .mu.M final concentration with one
equivalent of a complementary building block displaying a
nucleophilic recipient group. Reaction proceeds at temperatures
between 0.degree. C. and 100.degree. C. preferably between
15.degree. C.-50.degree. C. for 148 hours, preferably 10-20 hours
in DMF, DMSO, water, acetonitril, THF, DCM, methanol, ethanol or a
mixture thereof, pH buffered to 4-10, preferably 6-8. Organic
by-products are removed by extraction with EtOAc, followed by
evaporation of residual organic solvent for 10 min in vacuo. Pd
catalyst is removed and oligonucleotides are isolated by eluting
sample through a BiORad micro-spin chromatography column. Coupling
efficiency is quantified by ES-MS analysis.
[0212] General Procedure 7: Transfer of Functional Entity from a
Carrier Oligo to Recipient Reactive Group.
[0213] A carrier coupled functional entity oligo (Example 1)(250
pmol) was added to a scaffold oligo B (200 pmol) in 50 .mu.l 100 mM
MES, pH 6. The mixture was incubated overnight at 25.degree. C.
Subsequently, the mixture was purified by gel filtration using a
microspin column equilibrated with H.sub.2O and transfer of the
functional entity was verified by electron spray mass spectrometry
(ES-MS). Transfer efficiency is expressed in percent and were
calculated by dividing the abundance of scaffold oligo carrying
transferred functional entities to total abundance of scaffold
oligos (with and without transferred functional entities).
EXAMPLE 6
General Procedure 7
[0214] 18
[0215] Mass ("X"): 6583.97 (observed), 6583.31 (calculated).
Abundance: 65.79 (arbitrary units)
[0216] Mass ("Y"): 6599.73 (observed), 6597.34 (calculated).
Abundance: 29.23 (arbitrary units)
[0217] Mass ("Z"): 6789.36 (observed), 6790.65 (calculated)
[0218] Transfer efficiency calculated as: 29.23/ (29.23+65.79)
.dbd.0.3076.about.31%
[0219] General Procedure 8: Arylation of Oligonucleotide
Derivatives Containing a Nucleophilic Recipient Group Using a
Building Block of the Invention: 19
[0220] An oligonucleotide building block carrying functional entity
FE.sup.1 is combined at 2 .mu.M final concentration with one
equivalent of a complementary building block displaying a
nucleophilic recipient group. In the presence of a Pd catalyst, the
reaction proceeds at temperatures between 0.degree. C. and
100.degree. C. preferably between 15.degree. C.-50.degree. C. for
1-48 hours, preferably 10-20 hours in DMF, DMSO, water,
acetonitrile, THF, DCM, methanol, ethanol or a mixture thereof, pH
buffered to 4-10, preferably 6-8. Organic by-products are removed
by extraction with EtOAc, followed by evaporation of residual
organic solvent for 10 min in vacuo. Pd catalyst is removed and
oligonucleotides are isolated by eluting sample through a BiORad
micro-spin chromatography column. Coupling efficiency is quantified
by ES-MS analysis.
[0221] General Procedure 9: General Route to the Formation of
Alkylating/Vinylating Monomer Building Blocks with a
Thio-Succinimid S--C-Connecting Group and Use of These: 20
[0222] R.sup.1 and R.sup.2 may be used to tune the reactivity of
the sulphate to allow appropriate reactivity. Chloro and nitro
substitution will increase reactivity. Alkyl groups will decrease
reactivity. Ortho substituents to the sulphate will due to steric
reasons direct incoming nucleophiles to attack the R-group
selectively and avoid attack on sulphur. E.g. 21
[0223] 3-Aminophenol (6) is treated with maleic anhydride, followed
by treatment with an acid e.g. H.sub.2SO.sub.4 or P.sub.2O.sub.5
and heat to yield the maleimide (7). The ring closure to the
maleimide may also be achieved when an acid stable O-protection
group is used by treatment with or Ac.sub.2O with or without
heating, followed by O-deprotection. Alternatively reflux in
Ac.sub.2O, followed by O-deacetylation in hot water/dioxane to
yield (7).
[0224] Further treatment of (7) with SO.sub.2Cl.sub.2 with or
without triethylamine or potassium carbonate in dichloromethane or
a higher boiling solvent will yield the intermediate (8), which may
be isolated or directly further transformed into the aryl alkyl
sulphate by the quench with the appropriate alcohol, in this case
MeOH, whereby (9) will be formed. The organic building block (9)
may be connected to an oligo nucleotide, as follows.
[0225] A thiol carrying oligonucleotide in buffer 50 mM MOPS or
hepes or phosphate pH 7.5 is treated with a 1-100 mM solution and
preferably 7.5 mM solution of the organic building block (9) in
DMSO or alternatively DMF, such that the DMSO/DMF concentration is
5-50%, and preferably 10%. The mixture is left for 1-16 h and
preferably 24 h at 25.degree. C. To give the alkylating in this
case methylating monomer building block (10).
[0226] The reaction of the alkylating monomer building block (10)
with an amine carrying monomer building block may be conducted as
follows:
[0227] The coding oligonucleotide (1 nmol) is mixed with a thio
oligonucleotide loaded with a building block (1 nmol)(10) and an
amino-oligonucleotide (1 nmol) in hepes-buffer (20 .mu.L of a 100
mM hepes and 1 M NaCl solution, pH=7.5) and water (39 uL). The
oligonucleotides are annealed to the template by heating to
50.degree. C. and cooled (2.degree. C./second) to 30.degree. C. The
mixture is then left o/n at a fluctuating temperature (10.degree.
C. for 1 second then 35.degree. C. for 1 second), to yield the
template bound methylamine (11).
[0228] A vinylating monomer building block may be prepared and used
similarily as described above for an alkylating monomer building
block. Although instead of reacting the chlorosulphonate (8 above)
with an alcohol, the intermediate chlorosulphate is isolated and
treated with an enolate or O-trialkylsilylenolate with or without
the presence of fluoride. E.g. 22
[0229] Formation of the vinylating monomer building block (13):
[0230] The thiol carrying oligonucleotide in buffer 50 mM MOPS or
hepes or phosphate pH 7.5 is treated with a 1-100 mM solution and
preferably 7.5 mM solution of the organic building block (12) in
DMSO or alternatively DMF, such that the DMSO/DMF concentration is
5-50%, and preferably 10%. The mixture is left for 1-16 h and
preferably 2-4 h at 25.degree. C. To give the vinylating monomer
building block (13).
[0231] The sulfonylenolate (13) may be used to react with amine
carrying monomer building block to give an enamine (14a and/or 14b)
or e.g. react with an carbanion to yield (15a and/or 15b). E.g.
23
[0232] The reaction of the vinylating monomer building block (13)
and an amine or nitroalkyl carrying monomer building block may be
conducted as follows:
[0233] The coding oligonucleotide (1 nmol) is mixed with a
oligonucleotide building block (1 nmol)(13) and an
amino-oligonucleotide (1 nmol) or nitroalkyl-oligonucleotide (1
nmol) in 0.1 M TAPS, phosphate or hepes-buffer and 300 mM NaCl
solution, pH=7.5-8.5 and preferably pH=8.5. The oligonucleotides
are annealed to the template by heating to 50.degree. C. and cooled
(2.degree. C./second) to 30.degree. C. The mixture is then left o/n
at a fluctuating temperature (10.degree. C. for 1 second then
35.degree. C. for 1 second), to yield template bound (14a/b or
15a/b).
1 DCC N,N'-Dicyclohexylcarbodiimide DhbtOH
3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine DIC
Diisopropylcarbodiimide DIEA Diethylisopropylamin DMAP
4-Dimethylaminopyridine DNA Deoxyribosenucleic Acid EDC
1-Ethyl-3-(3'-dimethylaminopropyl)carbodiimide.HCl HATU
2-(1H-7-Azabenzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate HBTU 2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetrame-
thyluronium hexafluoro-phosphate HOAt N-Hydroxy-7-azabenzotriazole
HOBt N-Hydroxybenzotriazole LNA Locked Nucleic Acid NHS
N-hydroxysuccinimid OTf Trifluoromethylsulfonate OTs
Toluenesulfonate PNA Peptide Nucleic Acid PyBoP
Benzotriazole-1-yl-oxy-tris-pyrrolidino-phospho- nium
hexafluoro-phosphate PyBroP Bromo-tris-pyrrolidino-pho- sphonium
hexafluorophosphate RNA Ribonucleic acid TBTU
2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetra-
fluoroborate TEA Triethylamine RP-HPLC Reverse Phase High
Performance Liquid Chromatography TBDMS-Cl Tert-Butyldimethylsilyl-
chloride 5-Iodo-dU 5-iodo-deoxyriboseuracil TLC Thin layer
chromatography (Boc).sub.2O Boc anhydride, di-tert-butyl
dicarbonate TBAF Tetrabutylammonium fluoride SPDP
Succinimidyl-propyl-2-dithiopyridyl
* * * * *