US 7,351,413 B2 | ||
Stabilized HBc chimer particles as immunogens for chronic hepatitis | ||
Mark Page, Allestree (United Kingdom); Martin Friede, Nyon (Switzerland); Annette Elisabeth Schmidt, Planegg (Germany); and Detlef Stober, Munich (Germany) | ||
Assigned to Lorantis, Limited, Cambridge (United Kingdom) | ||
Filed on Oct. 01, 2003, as Appl. No. 10/677,074. | ||
Application 10/677074 is a continuation in part of application No. 10/372076, filed on Feb. 21, 2003, abandoned. | ||
Prior Publication US 2004/0156863 A1, Aug. 12, 2004 | ||
Int. Cl. A61K 39/00 (2006.01); A61K 39/12 (2006.01); A61K 39/29 (2006.01); A61K 39/39 (2006.01); A61K 9/107 (2006.01); C07K 14/00 (2006.01); C07K 14/02 (2006.01); C07K 14/005 (2006.01) |
U.S. Cl. 424—192.1 [424/184.1; 424/189.1; 424/204.1; 424/227.1; 424/278.1; 530/350] | 12 Claims |
1. A method of enhancing the production of one or more of gamma-producing CD 8+, CD 4+ T cells and cytotoxic T lymphocytes
against hepatitis B virus that comprises;
(a) administering to a patient chronically infected with hepatitis B virus a T cell-stimulating amount of a composition comprising
immunogenic particles dispersed in a squalene oil-in-water emulsion that includes an adjuvant comprised of a 2-[(R)-3-tetradecanoyloxytetra-decanoylamino]-ethyl-2-deoxy-4-O-phosphono-3-O-[(R)-3-tetradecanoyloxytetra-decanoyl]-2-[(R)-3-tetra-decanoyloxytetradecanoyl-amino]-p-D-glucopyranoside
triethylammonium salt, said immunogenic particles comprising recombinant hepatitis B core (HBc) chimeric protein molecules,
said chimeric protein molecules being up to about 550 amino acid residues in length and containing
(i) an HBc sequence of at least about 125 of the N-terminal 165 amino acid residues of the HBc molecule that includes the
HBc sequence of residue positions 4 through about 75 and about 85 through about 140, and includes an insert in the HBc immunodominant
loop, said insert having a length of one to about 40 amino acid residues containing one or more chemically non-reactive heterologous
amino acid residues that render the immunogenic particles less antigenic than the native HBc particles,
(ii) one or both of (a′) one to three cysteine residues at an amino acid position of the chimer molecule corresponding to
amino acid position −20 to about +1 from the N-terminus of the HBc sequence of SEQ ID NO:1 [N-terminal cysteine residue(s)]
in a sequence other than that of the HBc precore sequence and (b′) one to about three cysteine residues toward the C-terminus
of the molecule from the C-terminal residue of the HBc sequence and within about 30 residues from the C-terminus of the chimer
molecule [C-terminal cysteine residue(s)],
said chimer molecule (a′) containing no more than about 5 percent conservatively substituted amino acid residues in the HBc
sequence relative to one of SEQ ID NO:1-6 from position 2 through 165, (b′) having one or both HBc cysteine residues at positions
48 and 107 replaced by another residue, while the HBc cysteine at residue at position 61 is present, and (c′) self-assembling
into particles that upon expression in a host cell are substantially free of binding to nucleic acids, and
said particles being more stable than are particles formed from otherwise identical HBc chimer molecules that are free of
any above-mentioned C-terminal cysteine residue(s) or N-terminal cysteine residue(s) or in which a C-terminal or an N-terminal
cysteine residue(s) present in a contemplated chimer molecule is(are) replaced by another residue; and
(b) maintaining said patient for a time sufficient to induce T cells activated against HBc.
|