Adjuvant composition and methods for its use

Details Adjuvant composition and methods for its use Adjuvant composition and methods for its use

1999-05-07

2003-10-07

Minnifield, Nita (Department: 1645)

Drug, bio-affecting and body treating compositions

Preparations characterized by special physical form

Capsules

C424S450000, C424S283100, C424S690000, C424S689000

Reexamination Certificate

active

06630161

ABSTRACT:

BACKGROUND OF THE INVENTION
Vaccines comprise antigens or combinations of antigens which when administered to a warm-blooded animal prevent, ameliorate or treat disease. Vaccines for.infectious diseases originally comprised whole, attenuated or killed microbes. It was soon discovered however that only a few proteins or protein fragments of a microbe or cell stimulated a protective immune response, and, in fact, inclusion of extraneous materials from the whole cell could hinder the immune response. Therefore, vaccine development focused on identifying the particular protein, protein fragment, epitope and DNA segment encoding that epitope which elicited the protective immune response. As antigen identification became more precise however, vaccine efficiency declined. Identified: antigens were often small molecules unable to be recognized by antigen processing cells. It was therefore necessary to combine these antigens with substances which enhance the antigenicity of the antigen and give a superior immune response. These substances are adjuvants.
Adjuvants work by several means. Some assist in the presentation of antigen to antigen processing cells (APC). Oil-in-water emulsions, water-in-oil emulsions, liposomes and microbeads each assist in presenting antigen to APC. Small antigens or haptens are often linked to larger, immunogenic proteins or polysaccharides to facilitate recognition by the APC. Certain adjuvants have a depot effect holding antigen in place until the body has an opportunity to mount an immune response. Other adjuvants stimulate the immune system generally augmenting the specific response mounted to the antigen.
The attenuated lipid A derivatives (ALD) monophosphoryl lipid A (MLA) and 3-deacylated monophosphoryl lipid A (3D-MLA) are potent immunological adjuvants used in prophylactic vaccines for infectious disease and therapeutic vaccines for the treatment of cancerous tumors and chronic infections. MLA and 3D-MLA are modified forms of the bacterial endotoxin lipopolysaccharide (LPS) and are known and described in U.S. Pat. Nos. 4,436,727 and 4,912,094, respectively. MLA and 3D-MLA induce both a humoral antibody response and a cell-mediated immune response in patients administered the compounds with an antigen.
An effective vaccine presents antigens to a warm-blooded animal such that the animal can mount a protective immune response to those antigens. Often, a vaccine composition must include an adjuvant to achieve this effect. Adjuvants which stimulate both a humoral and cellular immune response and are safe and non-toxic would promote the efficacy of any vaccine.
SUMMARY OF THE INVENTION
The subject invention is a novel adjuvant composition. The adjuvant composition is a stable oil-in-water emulsion (SE) comprising a metabolizable oil, surfactants, an antioxidant and a component to make the emulsion isotonic. The particle size of the claimed stable emulsion is less than 130 nm to 3 &mgr;m. Emulsions in the range of 70-200 nm can be sterilized by filtration. The hydrophobic-lipophilic balance (HLB) of the stable emulsion.is from about 7.5 to about 10.5 and preferably about 8.0.
In a preferred embodiment, the adjuvant composition is combined with an attenuated lipid A derivative (ALD). The addition of an ALD increases the adjuvanticity of the composition. ALDs useful according to the subject invention include monophosphoryl lipid A and 3-deacylated monophosphoryl lipid A. ALD can be included in the formulation at a concentration ranging from about 1 &mgr;g-12,000 &mgr;g/ml. Vaccine compositions of the novel stable emulsion are also claimed.
DETAILED DESCRIPTION OF THE INVENTION
The subject invention is an adjuvant composition which is a stable oil-in-water emulsion comprising a metabolizable oil, surfactants, an antioxidant and a component to make the emulsion isotonic. The resulting emulsion is buffered, has a particle size of less than 3 &mgr;m and a hydrophobic-lipophilic balance of the stable emulsion is from about 7.5 to about 10.5 and preferably about 8.0.
In a preferred embodiment the stable emulsion comprises from about 2% to about 15%, and preferably 10%, volume/volume of the metabolizable oil squalene. Surfactants are present in the stable emulsion at about 2%. Approximately 50 &mgr;g of an antioxidant can be added to the stable emulsion of the subject invention and approximately 1.75% of an agent to make the emulsion isotonic.
Meabolizable oils useful according to the subject invention include squalene, soybean oil, sesame oil and caprylic/capric acid triglycerides (MIGLYCOL 810 oil). Squalene is preferred.
Surfactants useful according to the subject invention are Tween 80, polysorbate 80 (CAPMUL POE-O low PV surfactant, ABITEC Corp., Janesville, Wis.), polyethylene 660 12-hydroxystearate (SOLUTOL HS15, BASF Corp., Chicago, Ill.) and poloxamer 188 (PLURONIC Q F68 block co-polymer, BASF Corp., Chicago, Ill.), sodium cholate, glycerodeoxy cholate, phosphatidyl choline, with poloxamer 188 being preferred. It was found that Tween 80 and polysorbate 80 surfactant produced a histamine type response when administered intravenously to dogs. Other suitable surfactants include sphingolipids such as sphingomyelin and sphingosine and phospholipids such as egg phosphatidylcholine, 1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine, L-&agr;-Phosphatidylethanolamine, and 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or mixtures thereof. DPPC is acceptable for use in humans.
Antioxidants useful in the stable emulsion of the subject invention include a tocopherol, and ascorbic acid, with &agr; tocopherol being preferred.
Agents that can be added to the emulsion of the subject invention to make the adjuvant isotonic include dextrose, glycerol, mannitol, sorbitol, PEG 300, PEG 400 and polyethylene glycol, with glycerol being preferred.
In a particularly preferred embodiment, an attenuated lipid A derivative (ALD) is incorporated into the compositions of the subject invention. ALDs are lipid A-like molecules that have been altered or constructed so that the molecule displays lesser or different of the adverse effects of lipid A. These adverse effects include pyrogenicity, local Shwarzman reactivity and toxicity as evaluated in the chick embryo 50% lethal dose assay (CELD
50
) ALDs useful according to the subject invention include monophosphoryl lipid A (MLA) and 3-deacylated monophosphoryl lipid A (3D-7MLA). MLA and 3D-MLA are known and need not be described in detail herein. See for example U.S. Pat. No. 4,436,727 issued Mar. 13, 1984, assigned to Ribi ImmunoChem Research, Inc., which discloses monophosphoryl lipid A and its manufacture. U.S. Pat. No. 4,912,094 and reexamination certificate B1 U.S. Pat. No. 4,912,094 to Myers, et al., also assigned to Ribi ImmunoChem Research, Inc., embodies 3-deacylated monophosphoryl lipid A and a method for its manufacture. Disclosures of each of these patents with respect to MLA and 3D-MLA are incorporated herein by reference.
Without going into the details of the prior incorporated by reference patents, monophosphoryl lipid A (MLA) as used herein is derived from lipid A, a component of enterobacterial lipopolysaccharides (LPS), a potent but highly toxic immune system modulator. Edgar Ribi and his associates achieved the production of monophosphoryl lipid A (MLA) referred to originally as refined detoxified endotoxin (RDE). MLA is produced by refluxing an endotoxin extract (LPS or lipid A) obtained from heptoseless mutants of gram-negative bacteria in mineral acid solutions of moderate strength (0:1 N HCl) for a period of approximately 30 minutes. This treatment results in the loss of the phosphate moiety at position 1 of the reducing end glucosamine.
Coincidentally, the core carbohydrate is removed from the 6 position of the non-reducing glucosamine during this treatment. The resulting product (MLA) exhibits considerable attenuated levels of the endotoxic activities normally associated with the endotoxin starting material, such as pyrogenicity, local Shwarzman reactivity, and toxicity as evaluated in the chick embryo 50% leth

Affiliated with

Also associated with

Rating

Adjuvant composition and methods for its use does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Adjuvant composition and methods for its use, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Adjuvant composition and methods for its use will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3141638