Vaccine against
The present invention relates to a synthetic saccharide of general formula (I) that is related to Klebsiella pneumoniae serotype O1, O2, O2ac, and O8 O-polysaccharide and carbapenem-resistant Klebsiella pneumoniae ST258 O-polysaccharide and conjugate thereof. Said synthetic saccharide, said conjugate and pharmaceutical composition containing said synthetic saccharide or said conjugate are useful for prevention and/or treatment of diseases associated with Klebsiella pneumoniae . Furthermore, the synthetic saccharide of general formula (I) is useful as marker in immunological assays for detection of antibodies against Klebsiella pneumoniae bacteria.
1. A saccharide of general formula (II-4):
wherein
R 1 represents H or U 6 ,
U 6 represents
n is an integer from 1 to 20;
m is an integer from 1 to 20;
-L-E represents -L a -E, -L a -L e -E, -L a -L b -L e -E, or -L a -L d -L e -E;
wherein
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q CH 2 —,
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —, —CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
E represents —NH 2 , —N 3 , —O—NH 2 , —CH═CH 2 , —C≡CH, —Br, —Cl, —I, —COOH, —COOCH 3 , —COOC 2 H 5 , —CO 2 R′, —CO-(3-sulfo-N-hydroxysuccinimidyl), —CO-(dibenzocyclooctyne-sulfo-N-hydroxysuccinimidyl), —CONH—NH 2 , —OH, or —SH;
R′ represents
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6, with the proviso that -L-E is not —C 3 H 6 —NH 2 ;
or a pharmaceutically acceptable salt thereof.
2. The saccharide according to claim 1 , wherein n is an integer from 3 to 20,
or a pharmaceutically acceptable salt thereof.
3. The saccharide according to claim 2 , wherein n is an integer from 3 to 10,
m is 1 and R 1 is H;
or a pharmaceutically acceptable salt thereof.
4. The saccharide according to claim 3 , wherein -L- represents —(CH 2 ) o — and o is an integer selected from 4, 5 and 6;
or a pharmaceutically acceptable salt thereof.
5. The saccharide according to claim 1 , wherein the saccharide is selected from the group consisting of
or a pharmaceutically acceptable salt thereof.
6. The saccharide according to claim 1 , which is
or a pharmaceutically acceptable salt thereof.
7. A saccharide of general formula (II-8) or (II-11):
wherein
n is an integer from 1 to 20;
-L-E represents -L a -E, -L a -L e -E, -L a -L b -L e -E, or -L a -L d -L e -E;
wherein
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —;
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —, —CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
E represents —NH 2 , —N 3 , —O—NH 2 , —CH═CH 2 , —C≡CH, —Br, —Cl, —I, —COOH, —COOCH 3 , —COOC 2 H 5 , —CO 2 R′, —CO-(3-sulfo-N-hydroxysuccinimidyl), —CO-(dibenzocyclooctyne-sulfo-N-hydroxysuccinimidyl), —CONH—NH 2 , —OH, or —SH;
R′ represents
o, q, p1 and p2 are independently of each other an integer selected from 1,
2, 3, 4, 5, and 6, with the proviso that -L-E is not —C 3 H 6 —NH 2 ;
or a pharmaceutically acceptable salt thereof.
8. The saccharide according to claim 7 , wherein the saccharide is selected from the group consisting of
or a pharmaceutically acceptable salt thereof.
9. A conjugate of general formula (III-4):
wherein
R′ represents H or U 6 ,
U 6 represents
n is an integer from 1 to 20;
m is an integer from 1 to 20;
-L- represents -L a -, -L a -L e -, -L a -L b -L e -, or -L a -L d -L e -;
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —;
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —, —CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6;
E 1 - represents a covalent bond, —NH—, —O—NH—, —O—, —S—, —CO—,
—CH═CH—, —CONH—, —CO—NHNH—,
-T- represents
a represents an integer from 1 to 10;
b represents an integer from 1 to 4;
i is an integer selected from 2 to 25;
CP is a carrier protein selected from the group consisting of: a diphtheria toxoid, a mutated diphtheria toxoid, a modified diphtheria toxoid, a mutated and modified diphtheria toxoid, a tetanus toxoid, a modified tetanus toxoid, a mutated tetanus toxoid, outer membrane protein (OMP), bovine serum albumin (BSA), keyhole limpet hemocyanine (KLH), recombinant non-toxic form of Pseudomonas aeruginosa (rEPA) and cholera toxoid (CT).
10. The conjugate according to claim 3 , wherein n is an integer from 3 to 20.
11. The conjugate according to claim 10 , wherein n is an integer from 3 to 10, m is 1 and R 1 is H.
12. The conjugate according to claim 11 , wherein -L- represents —(CH 2 ) o — and o is an integer selected from 4, 5 and 6.
13. The conjugate according to claim 9 , wherein the conjugate is a conjugate of general formula (V-4)
wherein
R 1 represents H or U 6 ,
U 6 represents
n is an integer from 1 to 20;
m is an integer from 1 to 20;
-L- represents -L a -, -L a -L e -, -L a -L b -L e -, or -L a -L d -L e -;
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —;
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —,
—CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6;
-E 1 - represents a covalent bond, —NH—, —O—NH—, —O—, —S—, —CO—, —CH═CH—, —CONH—, —CO—NHNH—,
-T- represents
a represents an integer from 1 to 10;
b represents an integer from 1 to 4; and
i is an integer selected from 2 to 25.
14. The conjugate according to claim 13 , wherein
-T- represents
a is an integer selected from 2, 3, 4, 5, and 6; and
E 1 is a covalent bond, —NH—, —CH═CH—, —CONH—,
15. The conjugate according to claim 14 , wherein n is an integer from 3 to 20.
16. The conjugate according to claim 15 , wherein n is an integer from 3 to 10, m is 1 and R 1 is H.
17. The conjugate according to claim 16 , wherein -L- represents —(CH 2 ) o — and o is an integer selected from 4, 5 and 6.
18. The conjugate according to claim 17 , wherein i is an integer selected from 2 to 18.
19. The conjugate according to claim 13 , wherein i is an integer selected from 2 to 18.
20. The conjugate according to claim 18 , wherein i is an integer selected from 4 to 10.
21. A conjugate, wherein the conjugate comprises
wherein i is an integer selected from 2 to 25.
22. The conjugate according to claim 21 , wherein the conjugate comprises
wherein i is an integer selected from 2 to 18.
23. The conjugate according to claim 22 , wherein the conjugate comprises
wherein i is an integer selected from 4 to 10.
24. A conjugate having the formula:
25. A conjugate of general formula (III-8) or (III-11):
wherein
n is an integer from 1 to 20;
-L- represents -L a -L a -L e -, -L a -L b -L e -, or -L a -L d -L e -;
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —;
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —, —CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6;
-E 1 - represents a covalent bond, —NH—, —O—NH—, —O—, —S—, —CO—, —CH═CH—, —CONH—, —CO—NHNH—,
-T- represents
a represents an integer from 1 to 10;
b represents an integer from 1 to 4; and
i is an integer selected from 2 to 25;
CP is a carrier protein selected from the group consisting of: a diphtheria toxoid, a mutated diphtheria toxoid, a modified diphtheria toxoid, a mutated and modified diphtheria toxoid, a tetanus toxoid, a modified tetanus toxoid, a mutated tetanus toxoid, outer membrane protein (OMP), bovine serum albumin (BSA), keyhole limpet hemocyanine (KLH), recombinant non-toxic form of Pseudomonas aeruginosa (rEPA) and cholera toxoid (CT).
26. A pharmaceutical composition comprising at least one saccharide according to claim 1 as an active ingredient together with at least one pharmaceutically acceptable adjuvant and/or excipient.
27. A pharmaceutical composition comprising at least one conjugate according to claim 9 as an active ingredient together with at least one pharmaceutically acceptable adjuvant and/or excipient.
28. A method of inducing immune response against Klebsiella pneumoniae in a human or animal host, said method comprising administering at least one saccharide according to claim 1 to said human or animal host.
29. A method of inducing immune response against Klebsiella pneumoniae in a human or animal host, said method comprising administering at least one conjugate according to claim 9 to said human or animal host.
30. The method according to claim 28 , wherein the immune response is induced for the prevention and/or treatment of a disease associated with Klebsiella pneumoniae , wherein the disease is selected from meningitis, urinary tract infection, nosocomial pneumonia, intra-abdominal infections, wound infection, infection of blood, osteomyelitis, bacteremia, septicemia and ankylosing spondylitis.
31. The method according to claim 29 , wherein the immune response is induced for the prevention and/or treatment of a disease associated with Klebsiella pneumoniae , wherein the disease is selected from meningitis, urinary tract infection, nosocomial pneumonia, intra-abdominal infections, wound infection, infection of blood, osteomyelitis, bacteremia, septicemia and ankylosing spondylitis.
32. The method according to claim 30 , wherein the Klebsiella pneumoniae is selected from O-serotypes comprising or consisting of O1, O2a, O2ac, O2aeh, O2afg, O8, and carbapanem-resistant Klebsiella pneumoniae strain ST 258.
33. The method according to claim 31 , wherein the Klebsiella pneumoniae is selected from O-serotypes comprising or consisting of O1, O2a, O2ac, O2aeh, O2afg, O8, and carbapanem-resistant Klebsiella pneumoniae strain ST 258.
34. A process for preparing the conjugate of general formula (III-4) according to claim 9 , wherein the process comprises conjugating a saccharide of general formula (II-4):
wherein
R 1 represents H or U 6 ,
U 6 represents
n is an integer from 1 to 20;
m is an integer from 1 to 20;
-L-E represents -L a -E, -L a -L e -E, -L a -L b -L e -E, or -L a -L d -L e -E;
wherein
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —,
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —,
—CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
E represents —NH 2 , —N 3 , —O—NH 2 , —CH═CH 2 , —C≡CH, —Br, —Cl, —I, —COOH, —COOCH 3 , —COOC 2 H 5 , —CO 2 R′, —CO-(3-sulfo-N-hydroxysuccinimidyl), —CO-(dibenzocyclooctyne-sulfo-N-hydroxysuccinimidyl), —CONH—NH 2 , —OH, or —SH;
R′ represents
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6, with the proviso that -L-E is not —C 3 H 6 —NH 2 ;
or a pharmaceutically acceptable salt thereof,
to the carrier protein (CP).
35. A conjugate of general formula (III-4):
wherein
R 1 represents H or U 6 ,
U 6 represents
n is an integer from 1 to 20;
m is an integer from 1 to 20;
-L- represents -L a -, -L a -L e -, -L a -L b -L e , or -L a -L d -L e -;
-L a - represents —(CH 2 ) o —, —(CH 2 —CH 2 —O) o —C 2 H 4 —, or —(CH 2 —CH 2 —O) o —CH 2 ;
-L b - represents —O—, —NH—CO—NH—, —NH—CO—CH 2 —NH—, or —NH—CO—;
-L d - represents —(CH 2 ) q —, —(CH(OH)) q —, —(CF 2 ) q —, —(CH 2 —CH 2 —O) q —C 2 H 4 —, or —(CH 2 —CH 2 —O) q —CH 2 —;
-L e - represents —(CH 2 ) p1 —, —(CF 2 ) p1 —, —C 2 H 4 —(O—CH 2 —CH 2 ) p1 —, —CH 2 —(O—CH 2 —CH 2 ) p1 —, or —(CH 2 ) p1 —O—(CH 2 ) p2 —; and
o, q, p1 and p2 are independently of each other an integer selected from 1, 2, 3, 4, 5, and 6;
-E 1 - represents a covalent bond, —NH—, —O—NH—, —O—, —S—, —CO—, —CH═CH—, —CONH—, —CO—NHNH—,
-T- represents
a represents an integer from 1 to 10;
b represents an integer from 1 to 4;
i is an integer selected from 2 to 25; and
CP is a carrier protein selected from the group consisting of: a diphtheria toxoid, a mutated diphtheria toxoid, a modified diphtheria toxoid, a mutated and modified diphtheria toxoid, a tetanus toxoid, a modified tetanus toxoid, a mutated tetanus toxoid, outer membrane protein (OMP), bovine serum albumin (BSA), keyhole limpet hemocyanine (KLH), recombinant non-toxic form of Pseudomonas aeruginosa (rEP A) and cholera toxoid (CT),
wherein the conjugate is prepared by conjugating a saccharide of general formula (II-4):
to the carrier protein (CP).
36. A pharmaceutical composition comprising at least one conjugate according to claim 22 as an active ingredient together with at least one pharmaceutically acceptable adjuvant and/or excipient.
37. A method of inducing immune response against Klebsiella pneumoniae in a human or animal host, said method comprising administering at least one conjugate according to claim 22 to said human or animal host.
38. The method according to claim 37 , wherein the immune response is induced for the prevention and/or treatment of a disease associated with Klebsiella pneumoniae , wherein the disease is selected from meningitis, urinary tract infection, nosocomial pneumonia, intra-abdominal infections, wound infection, infection of blood, osteomyelitis, bacteremia, septicemia and ankylosing spondylitis.
39. The method according to claim 38 , wherein the Klebsiella pneumoniae is selected from O-serotypes comprising or consisting of O1, O2a, O2ac, O2aeh, O2afg, O8, and carbapanem-resistant Klebsiella pneumoniae strain ST 258.
40. A pharmaceutical composition, wherein the pharmaceutical composition comprises non-toxic mutated diphtheria toxin CRM 197 and the CRM 197 comprises at least one covalently bound saccharide as shown below: