Amide linkages of sugar moieties to amine terminated leashes attached to carbohydrate polymers
Provided are methods of attaching a mannose-binding C-type lectin receptor targeting moiety to a polymeric carbohydrate backbone using an amide linkage. The amide linkage may be found between a leash, such as an amine terminated leash, and the mannose-binding C-type lectin receptor targeting moieties. The compounds and compositions disclosed utilizing the amide linkage provide for highly stable compounds with a significant reduction in loss of mannose-binding C-type lectin receptor targeting moieties from the polymeric carbohydrate backbone.
1. A compound comprising:
a polymeric carbohydrate backbone comprising at least one amine terminated leash attached thereto, wherein the at least one amine terminated leash comprises a straight or branched chain having from about 1 to about 20 member atoms selected from the group consisting of carbon, oxygen, sulfur, nitrogen, and phosphorus; and
one or more mannose-binding C-type lectin receptor targeting moieties,
wherein the one or more mannose-binding C-type lectin receptor targeting moieties is attached to the at least one amine terminated leash as an amide linkage.
2. The compound of claim 1 , wherein the carbohydrate backbone comprises dextran, cellulose, mannan, chitin, or hyaluronic acid.
3. The compound of claim 1 , wherein the compound comprises a subunit as shown in Formula (I):
wherein
each X is independently H, L1-A, L1-Y-A, L2-R, or L3-H, wherein each X is bound to any OH group;
each of L1, L2, and L3 are independently leashes;
each A independently comprises a therapeutic agent, a diagnostic agent, or a theranostic agent;
each Y independently comprises a chelating agent;
each R independently comprises a mannose-binding C-type lectin receptor targeting moiety attached via a mannosyl coupling moiety; and
n is an integer greater than zero, wherein each unit of n may be the same or different;
wherein at least one R is present; and
wherein at least one L2 is the amine terminated leash to which the mannose-binding C-type lectin receptor targeting moiety is attached via a covalent amide linkage.
4. The compound of claim 3 , wherein each L2-R comprises the following formula:
wherein the bond 1A is attached to a hydroxide group of the polymeric carbohydrate backbone, wherein A is L2, wherein B is the mannosyl coupling moiety, wherein Cis R, and wherein y is an integer between 1 and 6.
5. The compound of claim 4 wherein B and C are attached to A via a precursor molecule of the following formula:
6. The compound of claim 4 wherein B and C are attached to A via an activated N-hydroxysuccinimide carboxylic acid ester.
7. The compound of claim 3 , wherein each of L1, L2, or L3 independently comprises —(CH 2 ) p S(CH 2 ) q —NH—, wherein p and q are integers from 0 to 5.
8. The compound of claim 3 , wherein L2 does not contain an amidine linkage.
9. The compound of claim 3 , wherein at least one subunit of formula (I) comprises:
10. The compound of claim 3 , wherein the compound comprises a subunit of formula (Ia):
wherein * indicates the point at which the therapeutic agent, diagnostic agent, or theranostic agent is attached, and wherein y is an integer between 1 and 6.
11. The compound of claim 1 , wherein the mannose-binding C-type lectin receptor targeting moiety comprises a mannosyl coupling reagent, mannose, high-mannose glycans or mannose oligosaccharides, fucose, N-acetylglucosamine, peptides, galactose, or a combination thereof.
12. The compound of claim 1 , wherein the polymeric carbohydrate backbone has a molecular weight of between about 1 kD to about 150 kD.
13. A pharmaceutical composition comprising:
the compound according to claim 1 ; and
a pharmaceutically acceptable carrier.
14. The composition of claim 13 , wherein the compound comprises a subunit as shown in Formula (I):
wherein
each X is independently H, L1-A, L1-Y-A, L2-R, or L3-H, wherein each X is bound to any OH group;
each of L1, L2, and L3 are independently leashes;
each A independently comprises a therapeutic agent, a diagnostic agent, or a theranostic agent;
each Y independently comprises a chelating agent;
each R independently comprises a mannose-binding C-type lectin receptor targeting moiety attached via a mannosyl coupling moiety; and
n is an integer greater than zero, wherein each unit of n may be the same or different;
wherein at least one R is present; and
wherein at least one L2 is the amine terminated leash to which the mannose-binding C-type lectin receptor targeting moiety is attached via a covalent amide linkage.
15. The composition of claim 14 , wherein each L2-R comprises the following formula:
wherein the bond 1A is attached to a hydroxide group of the polymeric carbohydrate backbone, wherein A is L2, wherein B is the mannosyl coupling moiety, wherein C is R, and wherein y is an integer between 1 and 6.
16. A method of attaching a mannose-binding C-type lectin receptor targeting moiety to a polymeric carbohydrate backbone, comprising:
(a) synthesizing a mannose-binding C-type lectin receptor targeting moiety comprising an anomeric thio-carboxylate moiety;
(b) converting the mannose-binding C-type lectin receptor targeting moiety comprising the anomeric thio-carboxylate moiety into an activated N-hydroxysuccinimide carboxylic acid ester; and
(c) reacting the activated N-hydroxysuccinimide carboxylic acid ester with a polymeric carbohydrate backbone having one or more amine terminated leashes attached thereto, wherein the at least one amine terminated leash comprises a straight or branched chain having from about 1 to about 20 member atoms selected from the group consisting of carbon, oxygen, sulfur, nitrogen, and phosphorus,
wherein the reacting of the activated N-hydroxysuccinimide carboxylic acid ester with the polymeric carbohydrate backbone having one or more amine terminated leashes attached thereto forms an amide linkage between the one or more amine terminated leashes and a carbonyl carbon of the activated N-hydroxysuccinimide carboxylic acid ester.
17. The method of claim 16 , wherein the method retains about 80% of the mannose-binding C-type lectin receptor targeting moieties on the polymeric carbohydrate backbone after about 20 hours.
18. The method of claim 16 , wherein the dextran backbone has a molecular weight between about 1 kD to about 150 kD.
19. The method of claim 16 , wherein the polymeric carbohydrate backbone having one or more amine terminated leashes attached thereto is formed before step (a) or at a point of time between step (a) and (c).