RECOMBINANT OBLIGATE ANAEROBIC GRAM-POSITIVE BACTERIA (As amended)

• US 20160326256A1
• Filed: 12/24/2014
• Published: 11/10/2016
• Est. Priority Date: 01/10/2014
• Status: Active Application

• Current Assignee: Teikyo Heisei University
• Original Assignee: Teikyo Heisei University
• Inventors: NISHIKAWA, Takeshi, TAIRA, Yuichiro, TAIRA, Ikuko, ISHIDA, Isao
• Granted Patent: US 10,266,601 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 35/74 Bacteria therapeutic use of a bacterial protein A61K38/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/569 Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 15/746 for lactic acid bacteria (Streptococcus; Lactococcus; Lactobacillus; Pediococcus; Enterococcus; Leuconostoc; Propionibacterium; Bifidobacterium; Sporolactobacillus)

Abstract

An object of the present invention is to effectively induce cancer cell apoptosis using the anti-TRAIL-R1 antibody(ies) and the anti-TRAIL-R2 antibody(ies) and to reduce the toxicity imposed on normal cells. The present invention relates to recombinant obligate anaerobic Gram-positive bacteria that include a nucleic acid encoding a fusion protein having 3 or more anti-TRAIL-R1 single-chain antibodies and/or 3 or more anti-TRAIL-R2 single-chain antibodies, in an expressible state.

Claims

1. Recombinant obligate anaerobic Gram-positive bacteria comprising a nucleic acid encoding a fusion protein comprising a signal peptide and 3 or more anti-TRAIL-R1 single-chain antibodies and/or 3 or more anti-TRAIL-R2 single-chain antibodies, in an expressible state.

2. The recombinant obligate anaerobic Gram-positive bacteria of claim 1, wherein the obligate anaerobic Gram-positive bacteria belong to the genus Bifidobacterium.

3. The recombinant obligate anaerobic Gram-positive bacteria of claim 1, wherein the anti-TRAIL-R1 single-chain antibody comprises CDR1 comprising the amino acid sequence as shown in SEQ ID NO:

22, CDR2 comprising the amino acid sequence as shown in SEQ ID NO;

23, and CDR3 comprising the amino acid sequence as shown in SEQ ID NO;

24.

4. The recombinant obligate anaerobic Gram-positive bacteria of claim 1, wherein the anti-TRAIL-R2 single-chain antibody comprises CDR1 comprising the amino acid sequence as shown in SEQ ID NO:

15, CDR2 comprising the amino acid sequence as shown in SEQ ID NO;

16, and CDR3 comprising the amino acid sequence as shown in SEQ ID NO;

17.

5. The recombinant obligate anaerobic Gram-positive bacteria of claim 1, wherein the fusion protein further comprises one or more functional peptides.

6. The recombinant obligate anaerobic Gram-positive bacteria of claim 5, wherein the functional peptides comprise a labeling protein.

7. An antitumor agent comprising, as an active ingredient, the recombinant obligate anaerobic Gram-positive bacteria of claim 1.

8. An anti-TRAIL-R1 antibody comprises CDR1 comprising the amino acid sequence as shown in SEQ ID NO:

22, CDR2 comprising the amino acid sequence as shown in SEQ ID NO;

23; and

CDR3 comprising the amino acid sequence as shown in SEQ ID NO;

24.