Fused cyclooctyne compounds and their use in metal-free click reactions

Patent metadata

Field	Value
Publication number	US-10239807-B2
Application number	US-201514978924-A
Country	US
Kind code	B2
Filing date	Dec 22, 2015
Priority date	Apr 27, 2010
Publication date	Mar 26, 2019
Grant date	Mar 26, 2019

Abstract

The invention relates to fused cyclooctyne compounds, and to a method for their preparation. The invention also relates to a conjugate wherein a fused cyclooctyne compound according to the invention is conjugated to a label, and to the use of these conjugates in bioorthogonal labeling, imaging and/or modification, such as for example surface modification, of a target molecule. The invention further relates to a method for the modification of a target molecule, wherein a conjugate according to the invention is reacted with a compound comprising a 1,3-dipole or a 1,3-(hetero)diene.

First claim

The invention claimed is:

1. A compound of the Formula (IIa), (IIb) or (IIc):

wherein:
n is 0 to 8;
p is 0 or 1;
R 3 is selected from the group consisting of [(L) p -Q], hydrogen, halogen, C 1 -C 24 alkyl groups, C 6 -C 24 (hetero)aryl groups, C 7 -C 24 alkyl(hetero)aryl groups and C 7 -C 24 (hetero)arylalkyl groups, the alkyl groups optionally being interrupted by one of more hetero-atoms selected from the group consisting of O, N and S, wherein the alkyl groups, (hetero)aryl groups, alkyl(hetero)aryl groups and (hetero)arylalkyl groups are independently optionally substituted with one or more substituents independently selected from the group consisting of C 1 -C 12 alkyl groups, C 2 -C 12 alkenyl groups, C 2 -C 12 alkynyl groups, C 3 -C 12 cycloalkyl groups, C 1 -C 12 alkoxy groups, C 2 -C 12 alkenyloxy groups, C 2 -C 12 alkynyloxy groups, C 3 -C 12 cycloalkyloxy groups, halogens, amino groups, oxo groups and silyl groups, wherein the alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, alkoxy groups, alkenyloxy groups, alkynyloxy groups and cycloalkyloxy groups are optionally substituted, the alkyl groups, the alkoxy groups, the cycloalkyl groups and the cycloalkoxy groups being optionally interrupted by one of more hetero-atoms selected from the group consisting of O, N and S, wherein the silyl groups are represented by the formula (R 4 ) 3 Si—, wherein R 4 is independently selected from the group consisting of C 1 -C 12 alkyl groups, C 2 -C 12 alkenyl groups, C 2 -C 12 alkynyl groups, C 3 -C 12 cycloalkyl groups, C 1 -C 12 alkoxy groups, C 2 -C 12 alkenyloxy groups, C 2 -C 12 alkynyloxy groups and C 3 -C 12 cycloalkyloxy groups, wherein the alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, alkoxy groups, alkenyloxy groups, alkynyloxy groups and cycloalkyloxy groups are optionally substituted, the alkyl groups, the alkoxy groups, the cycloalkyl groups and the cycloalkoxy groups being optionally interrupted by one of more hetero-atoms selected from the group consisting of O, N and S;
L is a linking group selected from linear or branched C 1 -C 24 alkylene groups, C 2 -C 24 alkenylene groups, C 2 -C 24 alkynylene groups, C 3 -C 24 cycloalkylene groups, C 5 -C 24 cycloalkenylene groups, C 8 -C 24 cycloalkynylene groups, C 7 -C 24 alkyl(hetero)arylene groups, C 7 -C 24 (hetero)arylalkylene groups, C 8 -C 24 (hetero)arylalkenylene groups, C 9 -C 24 (hetero)arylalkynylene groups, the alkylene groups, alkenylene groups, alkynylene groups, cycloalkylene groups, cycloalkenylene groups, cycloalkynylene groups, alkyl(hetero)arylene groups, (hetero)arylalkylene groups, (hetero)arylalkenylene groups and (hetero)arylalkynylene groups optionally being substituted with one or more substituents independently selected from the group consisting of C 1 -C 12 alkyl groups, C 2 -C 12 alkenyl groups, C 2 -C 12 alkynyl groups, C 3 -C 12 cycloalkyl groups, C 5 -C 12 cycloalkenyl groups, C 2 -C 12 cycloalkynyl groups, C 1 -C 12 alkoxy groups, C 2 -C 12 alkenyloxy groups, C 2 -C 12 alkynyloxy groups, C 3 -C 12 cycloalkyloxy groups, halogens, amino groups, oxo and silyl groups, wherein the silyl groups can be represented by the formula (R 4 ) 3 Si—, wherein R 4 is defined as above;
Q is a functional group selected from the group consisting of —CN, —N 3 , —NCX, —XCN, —XR 6 , —N(R 6 ) 2 , —+N(R 6 ) 3 , —C(X)N(R 6 ) 2 , —C(R 6 ) 2 XR 6 , —C(X)R 6 , —C(X)XR 6 , —XC(X)R 6 , —XC(X)XR 6 , —XC(X)N(R 6 ) 2 , —N(R 6 )C(X)R 6 , —N(R 6 )C(X)XR 6 and —N(R 6 )C(X)N(R 6 ) 2 , wherein X is oxygen or sulphur and wherein R 6 is independently selected from the group consisting of hydrogen, halogen, C 1 -C 24 alkyl groups, C 6 -C 24 (hetero)aryl groups, C 7 -C 24 alkyl(hetero)aryl groups and C 7 -C 24 (hetero)arylalkyl groups;
R 1 is independently selected from the group consisting of hydrogen, C 1 -C 24 alkyl groups, C 6 -C 24 (hetero)aryl groups, C 7 -C 24 alkyl(hetero)aryl groups and C 7 -C 24 (hetero)arylalkyl groups; and
R 2 is independently selected from the group consisting of halogen, —OR 6 , —NO 2 , —CN, —S(O) 2 R 6 , C 1 -C 12 alkyl groups, C 1 -C 12 aryl groups, C 1 -C 12 alkylaryl groups and C 1 -C 12 arylalkyl groups, wherein R 6 is as defined above, and wherein the alkyl groups, aryl groups, alkylaryl groups and arylalkyl groups are optionally substituted.

2. The compound according to claim 1 , wherein the compound is of the Formula (IIb) or (IIc).

3. The compound according to claim 1 , wherein p is 1 and L is CH 2 .

4. The compound according to claim 1 , wherein Q is selected from the group consisting of —OR 6 , —N(R 6 ) 2 , —+N(R 6 ) 3 , —C(O)N(R 6 ) 2 , —C(O)OR 6 , —OC(O)R 6 , —OC(O)OR 6 , —OC(O)N(R 6 ) 2 , —N(R 6 )C(O)R 6 , —N(R 6 )C(O)OR 6 and —N(R 6 )C(O)N(R 6 ) 2 , wherein R 6 is as defined in claim 1 .

5. The compound according to claim 1 , wherein Q is —OH.

6. The compound according to claim 1 , wherein R 1 is hydrogen.

7. The compound according to claim 1 , wherein R 3 is hydrogen or [(L) p -Q].

8. The compound according to claim 1 , wherein n is 0.

9. The compound according to claim 3 , wherein Q is —OH, R 1 is hydrogen, R 3 is hydrogen or [(L)-Q] and n is 0.

10. A conjugate comprising a compound according to claim 1 and a label, wherein the compound according to claim 1 is conjugated via Q to the label, wherein the label is selected from the group consisting of fluorophores, biotin, polyethylene glycol chains, polypropylene glycol chains, mixed polyethylene/polypropylene glycol chains, radioactive isotopes, steroids, pharmaceutical compounds, lipids, peptides, glycans, nucleotides and peptide tags.

11. A method for preparing a compound according to claim 1 , comprising:
(a) cyclopropanating a cyclooctadiene of the Formula (VIIa), (VIIb) or (VIIc):

wherein:
n=0 to 8;
R 1 is independently selected from the group consisting of hydrogen, C 1 -C 24 alkyl groups, C 6 -C 24 (hetero)aryl groups, C 7 -C 24 alkyl(hetero)aryl groups and C 7 -C 24 (hetero)arylalkyl groups; and
R 2 is independently selected from the group consisting of halogen, —OR 6 , —NO 2 , —CN, —S(O) 2 R 6 , C 1 -C 12 alkyl groups, C 1 -C 12 aryl groups, C 1 -C 12 alkylaryl groups and C 1 -C 12 arylalkyl groups, wherein the alkyl groups, aryl groups, alkylaryl groups and arylalkyl groups are optionally substituted, and wherein R 6 is independently selected from the group consisting of hydrogen, halogen, C 1 -C 24 alkyl groups, C 6 -C 24 (hetero)aryl groups, C 7 -C 24 alkyl(hetero)aryl groups and C 7 -C 24 (hetero)arylalkyl groups,
to form a bicyclic cyclooctene compound,
(b) brominating the obtained bicyclic cyclooctene compound to form a bicyclic cyclooctane compound, and
(c) dehydrobrominating the obtained bicyclic cyclooctane compound to form a compound according to claim 1 .

12. A method for producing a modified target molecule comprising reacting a conjugate according to claim 10 with a target molecule comprising a 1,3-dipole, a 1,3-diene, or a 1,3-heterodiene.

13. The method according to claim 12 , wherein the target molecule comprises a 1,3-dipole, selected from an azide, a nitrone or a nitrile oxide.

14. A composition comprising a conjugate according to claim 10 and a pharmaceutically acceptable carrier.

15. The compound according

Assignees

• Synaffix Bv

Inventors

• Van Delft Floris Louis
• Rutjes Floris Petrus Johannes Theodorus
• Dommerholt Frederik Jan

Classifications

• C07C2602/24

  the ring system containing nine carbon atoms, e.g. perhydroindane · CPC title

• C07C29/147

  of carboxylic acids or derivatives thereof · CPC title

• A61K49/0052

  Small organic molecules (oligomers, polymers, dendrimers A61K49/0054) · CPC title

• C07C33/16 Primary

  containing rings with more than six ring members · CPC title

• C07C269/06

  by reactions not involving the formation of carbamate groups · CPC title