JP2018523978A5

JP2018523978A5 -

Info

Publication number: JP2018523978A5
Application number: JP2017564379A
Authority: JP
Filing date: 2016-05-25
Publication date: 2019-06-20
Anticipated expiration: 2036-05-25

Claims

A genetically engineered bacterium comprising an arginine regulon, wherein the bacterium encodes a functional N-acetyl glutamic acid synthetase with lower arginine feedback inhibition compared to wild-type N-acetyl glutamic acid synthetase of the same bacterial subtype under the same conditions It includes a gene, the bacteria are genetically engineered to lack a functional ArgR, wherein the bacterium comprises a gene cassette encoding the biosynthetic pathway capable of producing short chain fatty acids, genetic engineering bacteria.

Biosynthetic pathway wherein said gene encoding arginine feedback resistant N-acetyl glutamic acid synthetase is operably linked to a promoter induced by exogenous environmental conditions and / or capable of producing short chain fatty acids The genetically engineered bacterium according to claim 1, wherein the gene cassette coding for is operably linked to a promoter induced by exogenous environmental conditions.

A promoter operably linked to the gene encoding functional N-acetyl glutamic acid synthetase with low arginine feedback inhibition and the gene cassette encoding a biosynthetic pathway capable of producing short chain fatty acids The genetically engineered bacterium according to claim 2, wherein the selected promoter is a separate copy of the same promoter.

The gene cassette encoding a biosynthetic pathway capable of producing short chain fatty acids acts on a promoter different from the promoter operably linked to the gene encoding functional N-acetyl glutamic acid synthetase with said low arginine feedback inhibition The genetically engineered bacterium according to claim 2 , which is linked as possible.

Promoter operably linked to a gene encoding a functional N- acetylglutamate synthetase having a low arginine feedback inhibition,
(A) induced by exogenous environmental conditions found in the small intestine of a mammal ;
(B) induced under hypoxic or anaerobic conditions; or
(C) induced by one or more molecules or metabolites indicative of liver damage,
The genetically engineered bacterium according to any one of claims 2 to 4.

A promoter operably linked to said gene cassette encoding a biosynthetic pathway capable of producing short chain fatty acids,
(A) induced by exogenous environmental conditions found in the small intestine of a mammal;
(B) induced under hypoxic or anaerobic conditions ;
(C) induced by one or more molecules or metabolites indicative of liver damage;
(D) induced by the presence of reactive nitrogen species;
(E) induced by the presence of reactive oxygen species; or
(F) induced by environmental factors not naturally present in the mammalian digestive tract,
The genetically engineered bacterium according to any one of claims 2 to 5 .

The gene encoding a functional N-acetyl glutamic acid synthetase having low arginine feedback inhibition and / or the gene cassette encoding a biosynthetic pathway capable of producing a short chain fatty acid can be used on chromosomes in bacteria or on plasmids in bacteria The genetically engineered bacterium according to any one of claims 1 to 6 , which is present in

5. Each copy of a functional argR gene normally present in the corresponding wild-type bacterium is independently deleted or rendered inactive by deletion, insertion or substitution of one or more nucleotides . 7. The genetically modified bacterium according to any one of 7 .

The genetically engineered bacterium according to any one of claims 1 to 8 , wherein the short chain fatty acid is butyric acid or propionic acid .

10. The genetically engineered bacterium according to any one of claims 1 to 9, which comprises resistance to rifaximin.

11. The probiotic bacterium according to any one of claims 1 to 10 , wherein the bacterium is a probiotic bacterium selected from the group consisting of Bacteroides, Bifidobacterium, Clostridia, E. coli, Lactobacillus and Lactococci. Bacteria described.

12. The bacterium according to claim 11 , wherein the bacterium is Escherichia coli nisrel.

The bacteria are
(A) the bacterium is an auxotrophic in the gene to be complemented if present in the mammalian gut ; and / or
(B) auxotrophs for enzymes in the diaminopimelic acid or thymine biosynthetic pathway,
The bacterium according to any one of claims 1 to 12 .

The bacterium is further engineered to carry an additional gene encoding a substance toxic to the bacterium, the additional gene being directly or indirectly induced by an environmental factor not naturally present in the mammalian digestive tract The bacterium according to any one of claims 1 to 13 , which is under the control of a promoter which

To any one of claims 1 to 14 including bacteria and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable compositions.

The composition according to claim 15, for use in a method of reducing hyperammonemia or treating a disease associated with hyperammonemia.

(A) the disease is a urea cycle disorder ;
(B) the disease is argininosuccinic aciduria, arginase deficiency, carbamoyl phosphate synthetase deficiency, citrullineemia, N-acetylglutamate synthetase deficiency or ornithine transcarbamylase deficiency;
(C) liver diseases; organic acid disorders; isovaleric acid uremia; 3-methylcrotonylglycineuria; methylmalonic aciduria; propionic aciduria; fatty acid oxidation defect; carnitine circuit defect; carnitine Deficiency; β-oxidation deficiency; lysine urinary protein intolerance; pyrroline-5-carboxylate synthetase deficiency; pyruvate carboxylase deficiency; ornithine aminotransferase deficiency; carbonic anhydrase deficiency; hyperinsulinism-high Ammoniaemia syndrome; Mitochondrial disorder; Valproic acid therapy; Asparaginase therapy; Complete parenteral nutrition; Cystoscopy with glycine-containing solution; Lung / bone marrow transplantation; Portal vein vein shunting; Urinary tract infection; Ureteral dilation Multiple myeloma; chemotherapy; infection; neurogenic bladder; or intestinal bacterial overgrowth; or
(D) the disease is hepatic encephalopathy, acute liver failure or chronic liver failure,
A composition for use according to claim 16 .