Treatment for cancer
AssigneeThe University of Nottingham
InventorsNigel Peter MINTON, John Timothy HEAP
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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 Km 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.
Claims60
A method of treatment for cancer in a subject comprising the administration to the subject of spores of 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 Km of less than 30 micromolar.
A method of preparing a subject for treatment with a prodrug for cancer comprising the administration to the subject of spores of 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 Km of less than 30 micromolar.
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 Km of less than 30 micromolar.
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 Km of less than 30 micromolar,
- for use as a medicament for treatment of a subject.
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 Km of less than 30 micromolar,
- for use in the preparation of a medicament for the treatment of cancer in a subject.
A prodrug for use as a medicament in a subject 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 Km of less than 30 micromolar.
The method or use according to any preceding claim, wherein a tumour of the subject is colonised with the obligate anaerobic microorganism.
The method or use according to any preceding claim, wherein the subject is colonised with the obligate anaerobic microorganism by administration of the obligate anaerobic microorganism, and/or spores thereof.
The method or use according to any preceding claim, wherein the subject is colonised with the obligate anaerobic microorganism by intravenous administration.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is capable of sporulation.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is administered in spore form.
The method or use according to any preceding claim, wherein at least about 5×107 spores of the obligate anaerobic microorganism are administered to the subject.
The method or use according to any preceding claim, wherein the cancer is a solid tumour cancer.
The method or use according to any preceding claim, wherein the polypeptide is capable of reducing CB1954 to the 4HX derivative with a Km of 25 micromolar or less.
The method or use according to any preceding claim, wherein the polypeptide is capable of reducing CB1954 to the 4HX derivative with a Km of 6.3 micromolar or less.
The method or use according to any preceding claim, wherein the polypeptide is capable of reducing CB1954 to a 4-hydroxylamine (4HX) derivative substantially without producing the 2-hydroxylamine derivative.
The method or use according to any preceding claim, 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);
- or a fragment, variant or derivative thereof.
The method or use according to any preceding claim, wherein the variant shares at least 20% sequence identify with the native polypeptide sequence.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is a bacterium.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is a bacterium of the class Clostridia.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is a Clostridial species selected from the group consisting of:
- c. Clostridium sporogenes;
- and
- d. Clostridium novyi.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism is auxotrophic for one or more essential nutrient.
The method or use according to any preceding claim, wherein the obligate anaerobic microorganism does not comprise an exogenous antibiotic resistance gene.
The method or use according to any preceding claim, wherein the polypeptide is encoded on the chromosome of the obligate anaerobic microorganism.
An isolated polypeptide having nitroreductase activity wherein the isolated polypeptide:
- e. exhibits preferential reduction of CB1954 to a 4-hydroxylamine (4HX) derivative instead of a 2-hydroxylamine derivative;
- and
- f. is capable of reducing CB1954 to the 4HX derivative with a Km of less than 30 micromolar.
An obligate anaerobic microorganism expressing or 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 Km of less than 30 micromolar.
An isolated polypeptide as defined in any one of claims 30 to 35, a polynucleotide as defined in any one of claims 36 to 38, a vector defined in any one of claims 39 to 41, an obligate anaerobic microorganism as defined in any one of claims 42 to 53 or a pharmaceutical composition as defined in claim 64 for use in directed enzyme prodrug therapy (DEPT);
- optionally, wherein the directed enzyme prodrug therapy is Clostridial directed enzyme prodrug therapy (CDEPT).
Use of an obligate anaerobic microorganism to convert a prodrug to a drug active in situ in a tumour, such as a solid tumour, wherein the obligate anaerobic microorganism is 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 Km of less than 30 micromolar.
Dependencies are inferred from the claim text.