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
Links
- 206010008631 Cholera Diseases 0.000 title claims 10
- 230000002265 prevention Effects 0.000 title claims 3
- 108010060123 Conjugate Vaccines Proteins 0.000 title 1
- 229940031670 conjugate vaccine Drugs 0.000 title 1
- 239000002596 immunotoxin Substances 0.000 title 1
- 229960005486 vaccine Drugs 0.000 claims abstract 23
- 241000607626 Vibrio cholerae Species 0.000 claims abstract 18
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims abstract 14
- 102000004169 proteins and genes Human genes 0.000 claims abstract 9
- 108090000623 proteins and genes Proteins 0.000 claims abstract 9
- 102000009016 Cholera Toxin Human genes 0.000 claims abstract 5
- 108010049048 Cholera Toxin Proteins 0.000 claims abstract 5
- 229960005004 cholera vaccine Drugs 0.000 claims abstract 3
- 239000000203 mixture Substances 0.000 claims abstract 2
- 230000002850 vibriocidal effect Effects 0.000 claims abstract 2
- 239000002158 endotoxin Substances 0.000 claims 17
- 229920006008 lipopolysaccharide Polymers 0.000 claims 17
- 238000000034 method Methods 0.000 claims 12
- 125000002252 acyl group Chemical group 0.000 claims 4
- 230000001588 bifunctional effect Effects 0.000 claims 4
- 235000014113 dietary fatty acids Nutrition 0.000 claims 4
- 239000003814 drug Substances 0.000 claims 4
- 229930195729 fatty acid Natural products 0.000 claims 4
- 239000000194 fatty acid Substances 0.000 claims 4
- 150000004665 fatty acids Chemical class 0.000 claims 4
- 230000003053 immunization Effects 0.000 claims 4
- GZQKNULLWNGMCW-PWQABINMSA-N lipid A (E. coli) Chemical compound O1[C@H](CO)[C@@H](OP(O)(O)=O)[C@H](OC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCCCC)[C@@H](NC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCC)[C@@H]1OC[C@@H]1[C@@H](O)[C@H](OC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](NC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](OP(O)(O)=O)O1 GZQKNULLWNGMCW-PWQABINMSA-N 0.000 claims 4
- 238000004519 manufacturing process Methods 0.000 claims 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims 2
- 229940032024 DPT vaccine Drugs 0.000 claims 2
- 201000005702 Pertussis Diseases 0.000 claims 2
- 150000008065 acid anhydrides Chemical class 0.000 claims 2
- 230000001580 bacterial effect Effects 0.000 claims 2
- 238000006243 chemical reaction Methods 0.000 claims 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 2
- 206010013023 diphtheria Diseases 0.000 claims 2
- 239000003937 drug carrier Substances 0.000 claims 2
- SPSXSWRZQFPVTJ-ZQQKUFEYSA-N hepatitis b vaccine Chemical compound C([C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC1N=CN=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)OC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@@H](N)CCCNC(N)=N)C1=CC=CC=C1 SPSXSWRZQFPVTJ-ZQQKUFEYSA-N 0.000 claims 2
- 229940124736 hepatitis-B vaccine Drugs 0.000 claims 2
- 244000005700 microbiome Species 0.000 claims 2
- 239000008194 pharmaceutical composition Substances 0.000 claims 2
- 230000001681 protective effect Effects 0.000 claims 2
- 229960000814 tetanus toxoid Drugs 0.000 claims 2
- JWDFQMWEFLOOED-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-(pyridin-2-yldisulfanyl)propanoate Chemical group O=C1CCC(=O)N1OC(=O)CCSSC1=CC=CC=N1 JWDFQMWEFLOOED-UHFFFAOYSA-N 0.000 claims 1
- 231100000699 Bacterial toxin Toxicity 0.000 claims 1
- 241000124008 Mammalia Species 0.000 claims 1
- 241000607598 Vibrio Species 0.000 claims 1
- IBVAQQYNSHJXBV-UHFFFAOYSA-N adipic acid dihydrazide Chemical compound NNC(=O)CCCCC(=O)NN IBVAQQYNSHJXBV-UHFFFAOYSA-N 0.000 claims 1
- 239000000688 bacterial toxin Substances 0.000 claims 1
- 230000001419 dependent effect Effects 0.000 claims 1
- NAQMVNRVTILPCV-UHFFFAOYSA-N hexane-1,6-diamine Chemical compound NCCCCCCN NAQMVNRVTILPCV-UHFFFAOYSA-N 0.000 claims 1
- 238000002649 immunization Methods 0.000 claims 1
- 238000001556 precipitation Methods 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 239000003053 toxin Substances 0.000 claims 1
- 231100000765 toxin Toxicity 0.000 claims 1
- 241000699802 Cricetulus griseus Species 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 1
- 238000005903 acid hydrolysis reaction Methods 0.000 abstract 1
- 230000000890 antigenic effect Effects 0.000 abstract 1
- 150000001720 carbohydrates Chemical class 0.000 abstract 1
- 229940030156 cell vaccine Drugs 0.000 abstract 1
- 230000002346 endotoxic effect Effects 0.000 abstract 1
- 238000009472 formulation Methods 0.000 abstract 1
- 230000001900 immune effect Effects 0.000 abstract 1
- 239000002510 pyrogen Substances 0.000 abstract 1
- 239000011780 sodium chloride Substances 0.000 abstract 1
- 230000002588 toxic effect Effects 0.000 abstract 1
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 |
|---|---|---|---|
| AU34696/93A AU678549B2 (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 |
| EP93903428A EP0623026A1 (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 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 |
|---|---|---|---|---|
| WO1998026799A1 (en) * | 1996-12-18 | 1998-06-25 | The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services | 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 |
| JP2004515450A (en) | 2000-04-18 | 2004-05-27 | エンドバイオロジックス， インコーポレイテッド | Lipopolysaccharide conjugate vaccine for the treatment of sepsis |
| US7749511B2 (en) | 2000-04-18 | 2010-07-06 | Endobiologics, Incorporated | Anti-sepsis conjugate vaccine |
| WO2002020059A2 (en) * | 2000-09-01 | 2002-03-14 | The Government Of The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Vibrio cholerae o139 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 |
| WO2003094961A1 (en) | 2002-05-09 | 2003-11-20 | Massimo Porro | Improved polysaccharide and glycoconjugate vaccines_____________ |
| WO2004043489A1 (en) * | 2002-11-14 | 2004-05-27 | Instituto Finlay. Centro De Investigacion-Produccion De Vacunas Y Sueros. | Method of obtaining conjugate vaccines and vaccine compositions containing same |
| 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 EP EP93903428A patent/EP0623026A1/en not_active Withdrawn
- 1993-01-14 CA CA002128212A patent/CA2128212A1/en not_active Abandoned
- 1993-01-14 AU AU34696/93A patent/AU678549B2/en not_active Ceased
- 1993-01-14 JP JP5512624A patent/JPH07503238A/en active Pending
- 1993-01-14 WO PCT/US1993/000253 patent/WO1993013797A2/en not_active Application Discontinuation
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