Method for preventing or treating M tuberculosis infection

• US 10,639,361 B2
• Filed: 06/25/2019
• Issued: 05/05/2020
• Est. Priority Date: 04/29/2005
• Status: Active Grant

• Current Assignee: Glaxosmithkline Biologicals SA (GlaxoSmithKline PLC)
• Original Assignee: Glaxosmithkline Biologicals SA (GlaxoSmithKline PLC)
• Inventors: Marchand, Martine
• Primary Examiner(s): Swartz, Rodney P
• Application Number:
  US16/451,040
• Publication Number:
  US 20190307872A1
• Time in Patent Office: 315 Days
• Field of Search: 4241841, 4241851, 4242341, 4242481, 536 231, 536 235
• CPC Class Codes: A61K 2039/523 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 2039/55572 Lipopolysaccharides; Lipid A; Monophosphoryl lipid A
• CPC Class Codes: A61K 2039/55577 Saponins; Quil A; QS21; ISCOMS
• CPC Class Codes: A61K 2039/575 humoral response
• CPC Class Codes: A61K 31/4409 only substituted in position 4, e.g. isoniazid, iproniazid
• CPC Class Codes: A61K 31/496 Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
• CPC Class Codes: A61K 39/04 Mycobacterium, e.g. Mycobacterium tuberculosis
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 31/06 for tuberculosis
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/35 from Mycobacteriaceae (F)

Abstract

The present invention is directed to methods of preventing reactivation of active and latent M. tuberculosis infections by administering a pharmaceutical composition comprising a nucleic acid encoding a Mtb72f fusion protein, or a Mtb72f fusion protein or an immunogenic fragment thereof, for example together with an adjuvant. The Mtb72f nucleic acid or fusion protein can be administered with one or more chemotherapeutic agents effective against a M. tuberculosis infection. The methods also provide for shortening the time course of a chemotherapeutic regimen against a M. tuberculosis infection.

Claims

1. A method of producing a polypeptide comprising the amino acid sequence of SEQ ID NO:

4, the method comprising;

(a) transforming or transfecting a host cell with an expression vector containing DNA encoding the polypeptide; and

(b) culturing the host cell under conditions suitable for the expression of the polypeptide from the host cell.

2. The method of claim 1, wherein the host cell is selected from the group consisting of a bacterial cell, a yeast cell, an insect cell, and an animal cell.

3. The method of claim 2, wherein the bacterial host cell is E. coli.

4. The method of claim 3, wherein the E. coli host cell is the HMS174 pLysS strain of E. coli.

5. The method of claim 2, wherein the host cell is a yeast cell.

6. The method of claim 2, wherein the animal host cell is a mammalian host cell.

7. The method of claim 6, wherein the mammalian host cell is selected from COS cell line, and CHO cell line.

8. The method of claim 1, further comprising the step of purifying the polypeptide from the cell culture.

9. The method of claim 8, wherein the step of purifying the polypeptide comprises at least one method selected from the group consisting of anion exchange chromatography, hydroxyapatite chromatography, diafiltration, and sterilizing filtration.

10. A method of producing a polypeptide comprising the amino acid sequence of SEQ ID NO:

4, the method comprising;

(a) transforming an E. coli host cell with an expression vector encoding the polypeptide;

(b) fermenting the transformed E. coli host cell of step (a) in a cell culture under conditions suitable for the expression of the protein from the E. coli host cell;

(c) harvesting the fermented E. coli host cell of step (b) by centrifugation of the cell culture;

(d) disrupting the harvested E. coli host cell of step (c);

(e) centrifuging the disrupted E. coli host cell of step (d) to produce an inclusion body pellet;

(f) extracting the inclusion body pellet of step (e) in a buffer comprising urea; and

(g) purifying the extracted inclusion body pellet of step (f) by anion exchange chromatography, hydroxyapatite chromatography, diafiltration, and sterilizing filtration.