WO2023128982A1 - A method for producing probiotic bacteria with magnetic characteristic that specifically targets cancer cells - Google Patents

A method for producing probiotic bacteria with magnetic characteristic that specifically targets cancer cells Download PDF

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WO2023128982A1
WO2023128982A1 PCT/TR2022/051032 TR2022051032W WO2023128982A1 WO 2023128982 A1 WO2023128982 A1 WO 2023128982A1 TR 2022051032 W TR2022051032 W TR 2022051032W WO 2023128982 A1 WO2023128982 A1 WO 2023128982A1
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magnetite
sequence
bacterial strain
magnetic resonance
probiotic bacteria
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PCT/TR2022/051032
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French (fr)
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Urartu Ozgur Safak SEKER
Merve YAVUZ
Ebru SAHIN KEHRIBAR
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Bilkent Universitesi Ulusal Nanoteknoloji Arastirma Merkezi
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Publication of WO2023128982A1 publication Critical patent/WO2023128982A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/26—Iron; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1896—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes not provided for elsewhere, e.g. cells, viruses, ghosts, red blood cells, virus capsides
    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0091—Oxidoreductases (1.) oxidizing metal ions (1.16)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y116/00—Oxidoreductases oxidizing metal ions (1.16)
    • C12Y116/03—Oxidoreductases oxidizing metal ions (1.16) with oxygen as acceptor (1.16.3)
    • C12Y116/03001—Ferroxidase (1.16.3.1), i.e. ceruloplasmin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/185—Escherichia
    • C12R2001/19—Escherichia coli

Definitions

  • the present invention relates to a method for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia.
  • Cancer is a prevalent disease on a global scale and cancer treatment and research have a significant share in the health sector.
  • magnetic resonance imaging techniques play a major role in the imaging and treatment targeting processes of patients. Since the image quality of magnetic resonance images depends on the tumor tissue’s apparent magnetic resonance response compared to the surrounding healthy tissues, the imaging process can become difficult in certain situations. Even if there are targeted magnetic markers used to enhance image clarity, marker delivery and metabolism pose a different challenge in the imaging process.
  • the United States patent document no. US10273469 discloses probiotic bacteria that deposit metal nanoparticles.
  • the said bacterial system can be used to improve mineral intake deficiency, to enhance image quality as a contrast agent in gastrointestinal imaging or to improve different tissue cancers by targeting cancer tissues, and magnetic nanoparticles are selectively deposited on the surface of selected probiotic bacteria in an appropriate pH environment.
  • the objective of the present invention is to realize a method for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia.
  • Figure l is a flowchart of the inventive method.
  • the inventive method (100) for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia comprises the steps of:
  • the step of determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101) included in the inventive method (100) is configured to determine at least one target probiotic bacteria strain that can colonize in the tumor region to be imaged and treated and does not cause septic shock in the patient's body.
  • the step of determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101) is configured to determine the target probiotic bacteria strain Escherichia coli BL21 (DE3).
  • the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) included in the inventive method (100) is configured to sequence at least one iron uptake protein, at least one iron retention protein, at least one controlled iron oxidation protein and at least one oxidized iron nucleation executive protein; to arrange a synthetic genetic circuit comprising the said sequences to produce the sequenced proteins; to transfer the synthetic genetic circuit to a target probiotic bacterium by at least one of horizontal gene transfer techniques.
  • the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the ion transporter protein and then insert it into synthetic genetic circuitry in order to facilitate the uptake of iron ions into the cell.
  • the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to insert the material binding peptide into synthetic genetic circuitry by sequencing it in order to increase its binding affinity to the iron oxide material.
  • the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the ferroxidase protein and then insert it into the synthetic genetic circuit in order to ensure efficient oxidation of ferrous ions with a positive charge of two.
  • the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the magnetite biomineralization protein, that has the characteristic of binding to iron and enables to nucleate the nanomaterial, and the inserts it into the synthetic genetic circuit.
  • the step of drugization of the magnetite-storing bacteria colony that exhibits the magnetite deposition sequence by multiplication (103) included in the inventive method (100) is configured to select the target probiotic bacteria that synthesize proteins by taking the synthetic genetic circuit, by directing the magnetites deposited in them with a magnetic field; to multiply the selected target probiotic bacteria at a concentration suitable for drug production; and to package the multiplied bacterial colony as a drug.
  • the step of ensuring colonization in tumor masses by taking the drugized magnetite-storing bacteria colony by the patient before the magnetic resonance imaging and the treatment of targeted magnetic resonance hyperthermia (104) included in the inventive method (100) is configured to administer the target probiotic bacteria containing magnetite to the patient as a drug; to allow the target probiotic bacteria colony to colonize within the tumor; to carry out contrast- enhanced magnetic resonance imaging transactions with the target probiotic bacteria colony; to create local hyperthermia within the tumor by keeping the target probiotic bacteria colony in the targeted magnetic resonance field; and to physically minimize the tumor tissue.
  • the target probiotic bacterial strain Escherichia coli BL21 (DE3) that can colonize in tumor masses and does not cause septic shock is determined (101); at least one iron uptake protein in the form of an iron transporter protein to facilitate the uptake of iron ions into the cell, and at least one iron retention protein in the form of a material binding peptide to increase its binding affinity of the iron oxide material, at least one controlled iron oxidation protein in the form of ferroxidase protein, and at least one oxidized iron nucleation executer protein in the form of magnetite biomineralization protein which binds to iron and enables nucleation of the nanomaterial to ensure effective oxidation of the plus divalent iron ions are sequenced and a synthetic genetic circuit containing these sequences to produce the sequenced proteins is arranged; and the synthetic genetic circuit is transferred to the target probiotic bacteria by at least one of the horizontal gene transfer techniques (102); the target probiotic bacteria that synthesize proteins by taking
  • the inventive method (100) enables to facilitate the transactions of magnetic resonance imaging and hyperthermia treatment by colonization of probiotic bacteria that spontaneously deposit magnetite in a tumor nutrient and oxygen environment.
  • inventive “Method for Producing Probiotic Bacteria with Magnetic Characteristic that Specifically Targets Cancer Cells (100)” the invention cannot be limited to examples disclosed herein and it is essentially according to claims.

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Abstract

The present invention relates to a method (100) for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia.

Description

A METHOD FOR PRODUCING PROBIOTIC BACTERIA WITH MAGNETIC CHARACTERISTIC THAT SPECIFICALLY TARGETS CANCER CELLS
Technical Field
The present invention relates to a method for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia.
Background of the Invention
Cancer is a prevalent disease on a global scale and cancer treatment and research have a significant share in the health sector. In cancer types wherein tumor masses are observed, magnetic resonance imaging techniques play a major role in the imaging and treatment targeting processes of patients. Since the image quality of magnetic resonance images depends on the tumor tissue’s apparent magnetic resonance response compared to the surrounding healthy tissues, the imaging process can become difficult in certain situations. Even if there are targeted magnetic markers used to enhance image clarity, marker delivery and metabolism pose a different challenge in the imaging process.
Considering the deficiencies of targeted magnetic markers in the current technique, there is a need for a method for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and targeted magnetic resonance hyperthermia therapy in the state of the art.
The United States patent document no. US10273469, an application in the state of the art, discloses probiotic bacteria that deposit metal nanoparticles. The said bacterial system can be used to improve mineral intake deficiency, to enhance image quality as a contrast agent in gastrointestinal imaging or to improve different tissue cancers by targeting cancer tissues, and magnetic nanoparticles are selectively deposited on the surface of selected probiotic bacteria in an appropriate pH environment.
Summary of the Invention
The objective of the present invention is to realize a method for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia.
Detailed Description of the Invention
“Method for Producing Probiotic Bacteria with Magnetic Characteristic that Specifically Targets Cancer Cells” realized to fulfil the objective of the present invention is shown in the figures attached, in which:
Figure l is a flowchart of the inventive method.
The components illustrated in the figures are individually numbered, where the numbers refer to the following: 100. Method
The inventive method (100) for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia comprises the steps of:
- determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101);
- inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102);
- drugization of the magnetite-storing bacteria colony that exhibits the magnetite deposition sequence by multiplication (103);
- ensuring colonization in tumor masses by taking the drugized magnetitestoring bacteria colony by the patient before the magnetic resonance imaging and the treatment of targeted magnetic resonance hyperthermia (104).
The step of determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101) included in the inventive method (100) is configured to determine at least one target probiotic bacteria strain that can colonize in the tumor region to be imaged and treated and does not cause septic shock in the patient's body. In one preferred embodiment of the invention, the step of determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101) is configured to determine the target probiotic bacteria strain Escherichia coli BL21 (DE3). The step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) included in the inventive method (100) is configured to sequence at least one iron uptake protein, at least one iron retention protein, at least one controlled iron oxidation protein and at least one oxidized iron nucleation executive protein; to arrange a synthetic genetic circuit comprising the said sequences to produce the sequenced proteins; to transfer the synthetic genetic circuit to a target probiotic bacterium by at least one of horizontal gene transfer techniques. In one preferred embodiment of the invention, the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the ion transporter protein and then insert it into synthetic genetic circuitry in order to facilitate the uptake of iron ions into the cell. In one preferred embodiment of the invention, the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to insert the material binding peptide into synthetic genetic circuitry by sequencing it in order to increase its binding affinity to the iron oxide material. In one preferred embodiment of the invention, the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the ferroxidase protein and then insert it into the synthetic genetic circuit in order to ensure efficient oxidation of ferrous ions with a positive charge of two. In one preferred embodiment of the invention, the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) is configured to sequence the magnetite biomineralization protein, that has the characteristic of binding to iron and enables to nucleate the nanomaterial, and the inserts it into the synthetic genetic circuit.
The step of drugization of the magnetite-storing bacteria colony that exhibits the magnetite deposition sequence by multiplication (103) included in the inventive method (100) is configured to select the target probiotic bacteria that synthesize proteins by taking the synthetic genetic circuit, by directing the magnetites deposited in them with a magnetic field; to multiply the selected target probiotic bacteria at a concentration suitable for drug production; and to package the multiplied bacterial colony as a drug.
The step of ensuring colonization in tumor masses by taking the drugized magnetite-storing bacteria colony by the patient before the magnetic resonance imaging and the treatment of targeted magnetic resonance hyperthermia (104) included in the inventive method (100) is configured to administer the target probiotic bacteria containing magnetite to the patient as a drug; to allow the target probiotic bacteria colony to colonize within the tumor; to carry out contrast- enhanced magnetic resonance imaging transactions with the target probiotic bacteria colony; to create local hyperthermia within the tumor by keeping the target probiotic bacteria colony in the targeted magnetic resonance field; and to physically minimize the tumor tissue.
Industrial Application of the Invention
In the inventive method (100), the In the method (100) of the present invention, the target probiotic bacterial strain Escherichia coli BL21 (DE3) that can colonize in tumor masses and does not cause septic shock is determined (101); at least one iron uptake protein in the form of an iron transporter protein to facilitate the uptake of iron ions into the cell, and at least one iron retention protein in the form of a material binding peptide to increase its binding affinity of the iron oxide material, at least one controlled iron oxidation protein in the form of ferroxidase protein, and at least one oxidized iron nucleation executer protein in the form of magnetite biomineralization protein which binds to iron and enables nucleation of the nanomaterial to ensure effective oxidation of the plus divalent iron ions are sequenced and a synthetic genetic circuit containing these sequences to produce the sequenced proteins is arranged; and the synthetic genetic circuit is transferred to the target probiotic bacteria by at least one of the horizontal gene transfer techniques (102); the target probiotic bacteria that synthesize proteins by taking the synthetic genetic circuit are selected by directing the magnetites deposited therein with a magnetic field; the selected target probiotic bacteria are multiplied at a concentration suitable for drug production, and the multiplied bacterial colony is packaged as a drug (103); the target probiotic bacteria containing magnetite are administered to the patient as a drug; colonization of the target probiotic bacteria colony within the tumor is ensured; contrast-enhanced magnetic resonance imaging procedures are performed with the target probiotic bacteria colony; local hyperthermia is created within the tumor by keeping the target probiotic bacteria colony in the targeted magnetic resonance field and the tumor tissue is physically minimized (104).
The inventive method (100) enables to facilitate the transactions of magnetic resonance imaging and hyperthermia treatment by colonization of probiotic bacteria that spontaneously deposit magnetite in a tumor nutrient and oxygen environment. Within these basic concepts; it is possible to develop various embodiments of the inventive “Method for Producing Probiotic Bacteria with Magnetic Characteristic that Specifically Targets Cancer Cells (100)”; the invention cannot be limited to examples disclosed herein and it is essentially according to claims.

Claims

CLAIMS A method (100) for enabling the deposition of magnetite in a probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock, and the use of magnetite-loaded probiotic bacteria in magnetic resonance imaging and treatment of targeted magnetic resonance hyperthermia; characterized by the step of:
- determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101);
- inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102);
- drugization of the magnetite-storing bacteria colony that exhibits the magnetite deposition sequence by multiplication (103);
- ensuring colonization in tumor masses by taking the drugized magnetite-storing bacteria colony by the patient before the magnetic resonance imaging and the treatment of targeted magnetic resonance hyperthermia (104). A method (100) according to Claim 1; characterized by the step of determining the target probiotic bacterial strain that can colonize in tumor masses and does not cause septic shock (101) which is configured to determine at least one target probiotic bacteria strain that can colonize in the tumor region to be imaged and treated and does not cause septic shock in the patient's body. A method (100) according to Claim 1 or 2; characterized by the step of determining the target probiotic bacterial strain that can colonize in tumor
8 masses and does not cause septic shock (101) which is configured to determine the target probiotic bacteria strain Escherichia coli BL21 (DE3). A method (100) according to any one of the preceding claims; characterized by the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) which is configured to sequence at least one iron uptake protein, at least one iron retention protein, at least one controlled iron oxidation protein and at least one oxidized iron nucleation executive protein; to arrange a synthetic genetic circuit comprising the said sequences to produce the sequenced proteins; to transfer the synthetic genetic circuit to a target probiotic bacterium by at least one of horizontal gene transfer techniques. A method (100) according to any one of the preceding claims; characterized by the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) which is configured to sequence the ion transporter protein and then insert it into synthetic genetic circuitry in order to facilitate the uptake of iron ions into the cell. A method (100) according to any one of the preceding claims; characterized by the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) which is configured to insert the material binding peptide into synthetic
9 genetic circuitry by sequencing it in order to increase its binding affinity to the iron oxide material. A method (100) according to any one of the preceding claims; characterized by the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) which is configured to sequence the ferroxidase protein and then insert it into the synthetic genetic circuit in order to ensure efficient oxidation of ferrous ions with a positive charge of two. A method (100) according to any one of the preceding claims; characterized by the step of inserting the sequence for enabling to produce proteins that will manage the intracellular uptake, intracellular retention, oxidation and nucleation of iron ions into the determined bacterial strain, in high volume as a magnetite deposition sequence (102) which is configured to sequence the magnetite biomineralization protein, that has the characteristic of binding to iron and enables to nucleate the nanomaterial, and the inserts it into the synthetic genetic circuit. A method (100) according to any one of the preceding claims; characterized by the step of drugization of the magnetite-storing bacteria colony that exhibits the magnetite deposition sequence by multiplication (103) which is configured to select the target probiotic bacteria that synthesize proteins by taking the synthetic genetic circuit, by directing the magnetites deposited in them with a magnetic field; to multiply the selected target probiotic bacteria at a concentration suitable for drug production; and to package the multiplied bacterial colony as a drug.
10 A method (100) according to any one of the preceding claims; characterized by the step of ensuring colonization in tumor masses by taking the drugized magnetite-storing bacteria colony by the patient before the magnetic resonance imaging and the treatment of targeted magnetic resonance hyperthermia (104) which is configured to administer the target probiotic bacteria containing magnetite to the patient as a drug; to allow the target probiotic bacteria colony to colonize within the tumor; to carry out contrast-enhanced magnetic resonance imaging transactions with the target probiotic bacteria colony; to create local hyperthermia within the tumor by keeping the target probiotic bacteria colony in the targeted magnetic resonance field; and to physically minimize the tumor tissue.
11
PCT/TR2022/051032 2021-12-29 2022-09-21 A method for producing probiotic bacteria with magnetic characteristic that specifically targets cancer cells WO2023128982A1 (en)

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TR2021/021659 2021-12-29

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130253303A1 (en) * 2012-01-13 2013-09-26 Bell Biosystems, Inc. Host Cells with Artificial Endosymbionts

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130253303A1 (en) * 2012-01-13 2013-09-26 Bell Biosystems, Inc. Host Cells with Artificial Endosymbionts

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PHILIP J. HILL, JOCHEN STRITZKER, MIRIAM SCADENG, ULRIKE GEISSINGER, DANIEL HADDAD, THOMAS C. BASSE-LüSEBRINK, UWE GBURECK, P: "Magnetic Resonance Imaging of Tumors Colonized with Bacterial Ferritin-Expressing Escherichia coli", PLOS ONE, vol. 6, no. 10, pages e25409, XP055649092, DOI: 10.1371/journal.pone.0025409 *
RAMESH P ET AL.: "Ultraparamagnetic Cells Formed through Intracellular Oxidation and Chelation of Paramagnetic Iron", ANGEW CHEM INT ED ENGL, vol. 57, no. 38, 17 September 2018 (2018-09-17), pages 12385 - 12389, XP055648363, [retrieved on 20180828], DOI: 10.1002/ anie. 20180504 2 *

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