Antimicrobial compositions comprising single domain antibodies and pseudomonas exotoxin

Patent metadata

Field	Value
Publication number	US-2016339112-A1
Application number	US-201515108380-A
Country	US
Kind code	A1
Filing date	Jan 2, 2015
Priority date	Jan 2, 2014
Publication date	Nov 24, 2016
Grant date	—

Abstract

The present invention provides heavy chain immunoglobulins of the VHH type or fragment thereof having affinity for a target antigen of interest, including glycoprotein D2 (gD2) of HSV-2 or antigen thereof, and for envelope proteins of HIV-1 or an antigen thereof linked to Pseudomonas exotoxin A or functional fragments thereof. Also included are multimeric forms of the immunoglobulins and their use in the prevention and/or treatment of HSV2 and/or HIV-1.

First claim

1 . (canceled)

2 . A heavy chain immunoglobulin of the VHH type or fragment thereof, wherein the heavy chain immunoglobulin of the VHH type or fragment thereof comprises an amino acid sequence of at least 90% identity to SEQ ID NO: 3 and having affinity for glycoprotein D2 (gD2) of HSV-2 or antigen thereof.

3 . The heavy chain immunoglobulin of the VHH type or fragment thereof of claim 2 , wherein the immunoglobulin or fragment thereof is covalently linked to the P. aeruginosa Exotoxin A subunit.

4 . The heavy chain immunoglobulin of the VHH type or fragment thereof of claim 3 , comprising an amino acid sequence of at least 90% identity to SEQ ID NO: 4.

5 . A multimeric molecule comprising a heavy chain immunoglobulin fragment of the VHH type according to claim 4 , in which VHH sequences are fused to yield multimeric units of 2 or more VHH units optionally linked via a spacer molecule.

6 . A multimeric molecule comprising two or more VHH sequences according to claim 4 , which are fused to yield 2, 3, 4 or 5 or more VHH units optionally linked via a spacer molecule.

7 . A nucleic acid encoding a heavy chain immunoglobulin fragment of the VHH type according to claim 2 .

8 . An expression vector comprising the gene encoding the heavy chain immunoglobulin fragment according to claim 7 .

9 . The expression vector of claim 8 comprising the nucleic acid sequence of SEQ ID NO: 4.

10 . A micro-organism transformed with the expression vector of claim 8 .

11 . The micro-organism according to claim 10 , wherein the micro-organism is Lactobacillus.

12 . The micro-organism according to claim 11 , selected from the group consisting of L. jensenii, L. reuteri, L. gasseri, L. crispatus , and L. iners.

13 . A method for treatment of an HSV2 infection in as subject, comprising administering to the subject, an effective amount of the compositions of claim 2 .

14 . A method for treatment of an HSV2 infection in as subject, comprising administering to the subject, an effective amount of a micro-organism of claim 12 expressing the heavy chain immunoglobulin or fragment thereof claim 2 .

15 . A heavy chain immunoglobulin of the VHH type or fragment thereof comprising an amino acid sequence of at least 85% identity to SEQ ID NO: 7, and having affinity for envelope proteins of HIV-1.

16 . A heavy chain immunoglobulin of the VHH type or fragment thereof comprising an amino acid sequence of at least 85% identity to SEQ ID NO: 8, and having affinity for envelope proteins of HIV-1, which is covalently linked to the P. aeruginosa Exotoxin A subunit.

17 . The heavy chain immunoglobulin of the VHH type or fragment thereof of claim 16 , having an amino acid sequence of at least 90% identity to SEQ ID NO: 8.

18 . A multimeric molecule comprising a heavy chain immunoglobulin fragment of the VHH type according to claim 16 , in which VHH sequences are fused to yield multimeric units of 2 or more VHH units optionally linked via a spacer molecule.

19 . A multimeric molecule comprising two or more VHH sequences according to claim 16 , which are fused to yield 2, 3, 4 or 5 or more VHH units optionally linked via a spacer molecule.

20 . A nucleic acid encoding a heavy chain immunoglobulin fragment of the VHH type according to claim 15 .

21 . An expression vector comprising the nucleic acid encoding a heavy chain immunoglobulin fragment of the VHH type according to claim 15 .

22 . The expression vector of claim 20 comprising the nucleic acid sequence of SEQ ID NO: 6.

23 . A micro-organism transformed with the expression vector of claim 22 .

24 . A micro-organism according to claim 23 , wherein the micro-organism is Lactobacillus.

25 . The micro-organism according to claim 24 , selected from the group consisting of L. jensenii, L. reuteri, L. gasseri, L. crispatus , and L. iners.

26 . A method for treatment of an HIV-1 infection in a subject, comprising administering to the subject a heavy chain immunoglobulin or fragment thereof of claim 15 .

27 . A method for treatment of an HIV-1 infection in a subject, comprising administering to subject a micro-organism of claim 23 expressing the heavy chain immunoglobulin or fragment of claim 15 .

28 . A method for treatment of an HSV2 infection in as subject, comprising administering to the subject, an effective amount of a micro-organism of claim 10 expressing the heavy chain immunoglobulin or fragment thereof of claim 2 , or the multimeric molecule of either of claim 5 or 6 .

29 . A method for treatment of an HSV2 infection in a subject, comprising administering to the subject, an effective amount of the multimeric molecule of claim 5 .

30 . A method for treatment of an HSV2 infection in as subject, comprising administering to the subject, an effective amount of a micro-organism of claim 12 expressing the multimeric molecule of either of claim 5 .

31 . A heavy chain immunoglobulin of the VHH type or fragment thereof comprising an amino acid sequence of at least 85% identity to SEQ ID NO: 11, and having affinity for envelope proteins of HIV-1.

32 . A heavy chain immunoglobulin of the VHH type or fragment thereof comprising an amino acid sequence of at least 85% identity to SEQ ID NO: 12, and having affinity for envelope proteins of HIV-1, which is covalently linked to the P. aeruginosa Exotoxin A subunit.

33 . The heavy chain immunoglobulin of the VHH type or fragment thereof of claim 32 , having an amino acid sequence of at least 90% identity to SEQ ID NO: 12.

34 . A method for treatment of an HIV-1 infection in a subject, comprising administering to the subject the multimeric molecule of claim 18 .

35 . A method for treatment of an HIV-1 infection in a subject, comprising administering to subject a micro-organism of claim 25 expressing the multimeric molecule of claim 19 .

36 . A method for treatment of an HSV2 infection in as subject, comprising administering to the subject, an effective amount of a micro-organism of claim 12 expressing the multimeric molecule of claim 6 .

Assignees

• Univ Johns Hopkins

Inventors

• Markham Richard
• Geoghegan Eileen

Classifications

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• A61K47/6839 Primary

  the antibody targeting material from viruses · CPC title

• C07K2317/35

  Valency · CPC title

• C07K2317/569

  Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® · CPC title

• A61K47/6829

  Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A · CPC title