Treatment for cancer

Patent metadata

Field	Value
Publication number	US-10183062-B2
Application number	US-201415038345-A
Country	US
Kind code	B2
Filing date	Nov 24, 2014
Priority date	Nov 22, 2013
Publication date	Jan 22, 2019
Grant date	Jan 22, 2019

Abstract

The present invention provides a method of treatment for cancer in a subject comprising the administration of a prodrug, wherein the subject is colonised with an obligate anaerobic microorganism capable of expressing a polypeptide having nitroreductase activity, wherein the polypeptide exhibits preferential reduction of CB1954 to a 4-hydroxylamine (4HX) derivative instead of a 2-hydroxylamine derivative; and is capable of reducing CB1954 to the 4HX derivative with a K m of less than 30 micromolar. The present invention further provides an isolated polypeptide, nucleic acids encoding said polypeptide, vectors and host cells comprising said nucleic acids and vectors and uses of the above in treating solid tumours.

First claim

The invention claimed is:

1. A method of treating a solid tumor cancer in a subject comprising the administration to the subject of an obligate anaerobic microorganism expressing a polypeptide having nitroreductase activity and/or the spores thereof, wherein the obligate anaerobic microorganism and/or spores thereof colonize a tumour of the subject, and wherein the polypeptide:
exhibits preferential reduction of CB1954 to a 4-hydroxylamine (4HX) derivative instead of a 2-hydroxylamine derivative; and
is capable of reducing CB1954 to the 4HX derivative with a K m of less than 30 micromolar; and

wherein the polypeptide comprises or consists of the amino acid sequence of:
a. SEQ ID NO: 1 (NmeNTR); or
b. SEQ ID NO: 2 (HsoNTR).

2. The method of claim 1 , further comprising the administration of a prodrug containing a nitro functional group to the subject.

3. The method according to claim 1 further comprising the administration of a prodrug comprising or consisting of CB1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide) or its analogue SN 23862 5-(N, N-bis(2-chloroethyl)amino)-2,4-dinitrobenzamide).

4. The method according to claim 1 , wherein the obligate anaerobic microorganisms and/or spores are administered to the subject by intravenous administration.

5. The method according to claim 1 , wherein the subject is colonised with the obligate anaerobic microorganism prior to administration of a prodrug containing a nitro functional group.

6. The method according to claim 1 , wherein the obligate anaerobic microorganism is capable of sporulation.

7. The method according to claim 1 , wherein the obligate anaerobic microorganism is administered in spore form.

8. The method according to claim 1 , wherein at least about 5×10 7 spores of the obligate anaerobic microorganism are administered to the subject.

9. The method according to claim 1 further comprising administering a prodrug containing a nitro functional group at a dose of at least 15 mg/kg.

10. The method according to claim 1 , wherein the polypeptide is capable of reducing CB1954 to the 4HX derivative with a K m of 25 micromolar or less.

11. The method according to claim 1 , wherein the polypeptide is capable of reducing CB1954 to a 4-hydroxylamine (4HX) derivative substantially without producing the 2-hydroxylamine derivative.

12. The method according to claim 1 , wherein the obligate anaerobic microorganism is a bacterium.

13. The method according to claim 1 , wherein the obligate anaerobic microorganism is a bacterium of the class Clostridia.

14. The method according to claim 1 , wherein the obligate anaerobic microorganism is a Clostridial species selected from the group consisting of:
c. Clostridium sporogenes ; and
d. Clostridium novyi .

15. The method according to claim 1 , wherein the obligate anaerobic microorganism is auxotrophic for one or more essential nutrient.

16. The method according to claim 1 , wherein the obligate anaerobic microorganism does not comprise an exogenous antibiotic resistance gene.

17. The method according to claim 1 , wherein the polypeptide is encoded on the chromosome of the obligate anaerobic microorganism.

18. A method of converting a prodrug to a drug active in situ in a tumour, such as a solid tumour in a subject in need thereof, comprising administering an obligate anaerobic microorganism and/or the spores thereof to the subject, wherein the obligate anaerobic microorganism and/or spores thereof expresses a polypeptide having nitroreductase activity, wherein the polypeptide:
exhibits preferential reduction of CB1954 to a 4-hydroxylamine (4HX) derivative instead of a 2-hydroxylamine derivative; and
is capable of reducing CB1954 to the 4HX derivative with a K m of less than 30 micromolar; and

wherein the polypeptide comprises or consists of the amino acid sequence of:
a. SEQ ID NO: 1 (NmeNTR); or
b. SEQ ID NO: 2 (HsoNTR).

Assignees

• Univ Nottingham

Inventors

• Minton Nigel Peter
• Heap John Timothy

Classifications

• C12Y105/01034

  6,7-Dihydropteridine reductase (1.5.1.34) · CPC title

• A61K35/742

  Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes · CPC title

• A61K31/396

  having three-membered rings, e.g. aziridine · CPC title

• C12N9/0028

  with NAD or NADP as acceptor (1.5.1) · CPC title

• A61K35/745

  Bifidobacteria · CPC title