WO1993013797B1 - Detoxified lps-cholera toxin conjugate vaccine for prevention of cholera - Google Patents
Detoxified lps-cholera toxin conjugate vaccine for prevention of choleraInfo
- Publication number
- WO1993013797B1 WO1993013797B1 PCT/US1993/000253 US9300253W WO9313797B1 WO 1993013797 B1 WO1993013797 B1 WO 1993013797B1 US 9300253 W US9300253 W US 9300253W WO 9313797 B1 WO9313797 B1 WO 9313797B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lipopolysaccharide
- cholera
- vaccine
- detoxified
- protein carrier
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
Abstract
A vaccine formulation comprising conjugates of detoxified LPS with proteins including cholera toxin (CT) is disclosed. Treatment with hydrazine (DeA-LPS) reduces the endotoxic properties of the LPS to clinically acceptable levels and results in a larger and more antigenic molecule than the saccharide produced by acid hydrolysis. The conjugates utilizing the cholera toxin of V. cholerae are disclosed which have low levels of pyrogen, no toxic activity upon Chinese hamster overay cells and elicit booster responses of vibriocidal and CT antibodies when injected subcutaneously as saline solutions into mice. The conjugates produced as a cholera vaccine induce the same antibodies as parenterally injected cellular vaccines but have improved safety and immunologic properties.
Claims
1. A detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate comprising the detoxified lipopolysaccharide of Vibrio cholera, said lipopolysaccharide detoxified using anhydrous hydrazine, covalently attached to a protein carrier by means of a bifunctional linker to form a detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate.
2. A cholera vaccine comprising: the detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate according to claim 1.
3. A cholera vaccine comprising: the detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate according to claim 1, wherein the anhydrous hydrazine selectively removes acyl linked fatty acids from a lipid A component of lipopolysaccharide.
4. A vaccine according to claim 2 wherein the protein carrier is isolated from or secreted by a bacterial strain.
5. A vaccine according to claim 4, wherein said protein carrier is a secreted protein.
6. A vaccine according to claim 4, wherein said protein carrier is a bacterial toxin.
7. A vaccine according claim 6, wherein said toxin is the cholera toxin of Vibrio cholera.
8. A vaccine according to claim 2, wherein said covalent attachment is accomplished by reaction with a bifunctional linker selected from the group consisting of adipic acid dihydrazide, diaminohexane, amino- e-caproic acid, and an N-hydrosuccinimide acid anhydride-based heterobifunctional linker.
9. A vaccine according to claim 8, wherein said N- hydrosuccinimide acid anhydride-based heterobifunctional linker is N-succinimidyl- 3-(2-pyridyldithio) propionate.
10. A vaccine of claim 2, wherein the conjugate is formed between a detoxified derivatized LPS and a protein carrier by reaction with 1-ethyl- 3 (3-dimethylaminopropyl) carbodiimide.
11. A method for preparing a covalently linked detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate comprising:
1) detoxifying Vibrio cholera lipopolysaccharide using anhydrous hydrazine under conditions that selectively remove acyl linked fatty acids from the lipid A component,
2) reacting the detoxified Vibrio cholera lipopolysaccharide of step 1 with a bifunctional linker,
3) mixing the derivatized, detoxified Vibrio cholera lipopolysaccharide from step (2) with a protein carrier,
4) adding to the mixture of step (3) a reagent, said reagent causing covalent linkage to occur to form the conjugate.
12. A vaccine according to claim 7, which further comprises a second vaccine directed against a second microorganism.
13. A vaccine according to claim 12, wherein said second vaccine is selected from the group consisting of diphtheria (D), tetanus toxoid (T), pertussis (P), DTP, and Hepatitis B vaccine.
14. A vaccine comprising the conjugate of claim 4, and a second vaccine directed against a second microorganism.
15. The vaccine of claim 14, wherein said second vaccine is selected from the group consisting of diphtheria (D), tetanus toxoid (T), pertussis (P), DTP, and Hepatitis B vaccine.
16. Use of a vaccine according to claim 2 for the manufacture of a medicament for use in a method for immunizing a human against cholera which comprises administering an amount of the vaccine of claim 2, sufficient to provide protection against cholera to the human.
17. Use of a vaccine according to claim 4 for the manufacture of a medicament for use in a method for immunizing „. human against cholera which comprises administering an amount of the vaccine c claim 4, sufficient to provide protection against cholera to the human.
18. Use of a vaccine according to claim 7 for the manufacture of a medicament for use in a method for immunizing a human against cholera which comprises administering an amount of the conjugate of claim 7, sufficient to provide protection against cholera, to the human.
19. A method for detoxifying Vibrio cholera lipopolysaccharide and retaining protective antigenicity comprising the steps:
A) treating the lipopolysaccharide with an effective amount of anhydrous hydrazine to selectively remove acyl linked fatty acids from the lipid A component.
20. A method according to claim 19 wherein step (A) is performed at a temperature of about 37°C.
21. A method according to claim 19 wherein step (A) is performed for about 120 minutes.
22. A method according to claim 19 further comprising step (B) purifying the lipopolysaccharide.
23. A method according to claim 22 step (B) wherein the purification is performed by precipitation of the detoxified lipopolysaccharide using about 90% acetone at about 4°C.
24. A purified and isolated antibody produced by a mammal in response to immunization with the vaccine according to claim 2, 4, or 7 said antibody characterized in that it reacts with lipopolysaccharide on Vibrio cholera and has vibriocidal activity.
25. A purified and isolated antibody of claim 24 wherein the antibody is polyclonal.
26. A pharmaceutical composition comprising the antibody according to claim 24 and a pharmaceutically acceptable carrier.
27. Use of an antibody of claim 24 for the manufacture of a medicament for use in a method for prevention or treatment of cholera in a human, comprising: administration of the antibodies in an amount effective for prevention or treatment of cholera.
28. A method of claim 11 further comprising: Step (5) isolating the detoxified Vibrio cholera lipopolysaccharide-protein conjugate.
29. A pharmaceutical composition comprising the vaccine according to claims 2, 4, or 7 and a pharmaceutically acceptable carrier. STATEMENT UNDER ARTICLE 19
In response to the Notification of Transmittal of the International Search Report or Declaration dated September 22, 1993 and in reply to the communication dated November 29, 1993, Applicant is forwarding herein replacements sheets of amended claims.
New independent Claim 1 differs from the original Claim 1 in that it recites a detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate in place of detoxified bacterial LPS. New Claim 1 also recites that the Vibrio chc ~ lipopolysaccharide is detoxified using anhydrous hydrazine and that the detoxif LPS is covalently attached to the protein carrier by a bifunctional linker.
Original claim 13 was amended and renumbered as claim 11. Claim 11 differs from the original claim 13 in the recitation of detoxified Vibrio cholera lipopolysaccharide and in the recitation of anhydrous hydrazine and the recitation of selective removal of acyl linked fatty acids from the lipid A component.
New independent claim 19 and dependent claims 20-23 relate to a method of detoxifying Vibrio cholera lipopolysaccharide and retaining protective ■antigenicity.
New claims 24-27 relate to antibody produced in response to the detoxified Vibrio cholera lipopolysaccharide-protein carrier conjugate.
Applicant respectfully submits that the amendment does not introduce new matter and are fully supported by the description and drawings.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP93903428A EP0623026A1 (en) | 1992-01-16 | 1993-01-14 | Detoxified lps-cholera toxin conjugate vaccine for prevention of cholera |
| JP5512624A JPH07503238A (en) | 1992-01-16 | 1993-01-14 | Detoxified LPS-cholera toxin conjugate vaccine for cholera prevention |
| AU34696/93A AU678549B2 (en) | 1992-01-16 | 1993-01-14 | Detoxified LPS-cholera toxin conjugate vaccine for prevention of cholera |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US82145392A | 1992-01-16 | 1992-01-16 | |
| US07/821,453 | 1992-01-16 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO1993013797A2 WO1993013797A2 (en) | 1993-07-22 |
| WO1993013797A3 WO1993013797A3 (en) | 1993-10-28 |
| WO1993013797B1 true WO1993013797B1 (en) | 1994-01-20 |
Family
ID=25233450
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US1993/000253 Ceased WO1993013797A2 (en) | 1992-01-16 | 1993-01-14 | Detoxified lps-cholera toxin conjugate vaccine for prevention of cholera |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0623026A1 (en) |
| JP (1) | JPH07503238A (en) |
| AU (1) | AU678549B2 (en) |
| CA (1) | CA2128212A1 (en) |
| WO (1) | WO1993013797A2 (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU1420897A (en) * | 1996-12-18 | 1998-07-15 | Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services, The | Conjugate vaccine for (salmonella paratyphi) a |
| US6531131B1 (en) | 1999-08-10 | 2003-03-11 | The United States Of America As Represented By The Department Of Health And Human Services | Conjugate vaccine for Neisseria meningitidis |
| US7749511B2 (en) | 2000-04-18 | 2010-07-06 | Endobiologics, Incorporated | Anti-sepsis conjugate vaccine |
| EP1278548A2 (en) | 2000-04-18 | 2003-01-29 | Endobiologics, Incorporated | Lipopolysaccharide-conjugate vaccine for sepsis treatment |
| US7527797B1 (en) | 2000-09-01 | 2009-05-05 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Vibrio cholerae 0139 conjugate vaccines |
| WO2002080964A1 (en) * | 2001-04-06 | 2002-10-17 | Institut Pasteur | Conjugate vaccine composed of the polysaccharide moiety of the lipopolysaccharide of vibrio cholerae 0139 bound to tetanus toxoid |
| AU2002309259A1 (en) * | 2002-05-09 | 2003-11-11 | Massimo Porro | Improved polysaccharide and glycoconjugate vaccines_____________ |
| BR0316271A (en) * | 2002-11-14 | 2005-10-11 | Inst Finlay Ct De Investigacio | Method for obtaining multivalent conjugate vaccine preparations, multivalent vaccine composition and use thereof |
| US8048432B2 (en) | 2003-08-06 | 2011-11-01 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Polysaccharide-protein conjugate vaccines |
| CN102824632A (en) * | 2012-09-12 | 2012-12-19 | 北京民海生物科技有限公司 | Polysaccharide conjugate vaccine of vibrio cholera group O1, preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2532850B1 (en) * | 1982-09-15 | 1985-12-20 | Pasteur Institut | IMMUNOGENIC CONJUGATES BETWEEN A HAPTENA AND A CARRIER MOLECULE DERIVED FROM A TOXIN, THE VACCINES CONTAINING THEM AND PROCESS FOR OBTAINING THEM |
| US4663160A (en) * | 1983-03-14 | 1987-05-05 | Miles Laboratories, Inc. | Vaccines for gram-negative bacteria |
-
1993
- 1993-01-14 WO PCT/US1993/000253 patent/WO1993013797A2/en not_active Ceased
- 1993-01-14 AU AU34696/93A patent/AU678549B2/en not_active Ceased
- 1993-01-14 EP EP93903428A patent/EP0623026A1/en not_active Withdrawn
- 1993-01-14 JP JP5512624A patent/JPH07503238A/en active Pending
- 1993-01-14 CA CA002128212A patent/CA2128212A1/en not_active Abandoned
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