FR3142904A1 - NANOPARTICLES ARRANGED IN CHAINS IN AN ISOTONIC COMPOSITION AFTER RECONSTITUTION - Google Patents

Abstract

NANOPARTICLES ARRANGED IN CHAINS IN AN ISOTONIC COMPOSITION AFTER RECONSTITUTION The invention relates to a composition comprising at least one chain of at least two nanoparticles, in which each nanoparticle of the chain comprises a mineral core of iron oxide surrounded by a coating, wherein the composition further comprises a cryoprotectant, wherein the dissociation energy between the coating and the core is greater than the dissociation energy between the cryoprotectant and the core, wherein the chain arrangement is maintained at a temperature of the composition which is less than 0°C. Figure for the abstract: No figure

Description

NANOPARTICLES ARRANGED IN CHAINS IN AN ISOTONIC COMPOSITION AFTER RECONSTITUTION FIELD OF THE INVENTION

The field of the invention is that of nanoparticles, presenting a particular type of organization, such as an organization in chains, the organization of which can be maintained for a long period, in particular to preserve the chains, by lyophilizing these nanoparticles in presence of a cryoprotectant, by maintaining these nanoparticles lyophilized for a certain time, and by resuspending the chains in a liquid such as water, preferably before their use or administration to humans.

BACKGROUND OF THE INVENTION

Some nanoparticles are known to organize themselves in a specific way. For example, magnetosomes synthesized by magnetotactic bacteria which consist of iron oxide mineral cores covered with organic lipid bilayers form chains, (Applications of magnetotactic bacteria and the magnetosome for cancer treatment: A review emphasizing on practical and mechanistic aspects, Edouard Alphandéry, Drug Discovery Today, V. 25, P. 1444, 2020). We were able to remove the lipid bilayer covering the mineral core of the magnetosomes, replacing it with a synthetic coating (see reference above). However, the synthetic coating of the mineral core of magnetosomes degrades in water over time. These nanoparticles cannot be maintained in a suspended chain arrangement over a long period of time. Here we offer a new product consisting of chains of magnetosomes mixed with a cryoprotectant, which are freeze-dried to be stored in powder form over a long period. They are then resuspended in a liquid, where they recompose into chains, before being used.

DESCRIPTION OF THE INVENTION

The invention relates to a composition comprising at least one chain of at least two nanoparticles, wherein each nanoparticle of the chain comprises an iron oxide mineral core or an iron oxide core or a metal core or a compound core. a metal oxide, surrounded by a coating,
in which the composition further comprises a cryoprotectant which is optionally mixed with water,
in which the dissociation energy between the coating and the core is preferably greater than the dissociation energy between the cryoprotectant and the core,
in which the composition is preferably in the form of a powder or a liquid suspension,
in which the coating preferentially maintains the arrangement of the chains above 0°C,
in which the cryoprotective agent preferentially maintains the arrangement of the chains below 0°C,
wherein the chain arrangement is preferably maintained at a temperature below 0°C.

The invention also relates to a composition comprising at least two nanoparticles organized in a geometric figure, in which each nanoparticle of the geometric figure comprises an iron oxide mineral core surrounded by a coating,
in which the composition further comprises a cryoprotectant preferably mixed with water,
in which the dissociation energy between the coating and the core is preferably greater than the dissociation energy between the cryoprotectant and the core,
in which the cryoprotective agent preferably serves to maintain the geometric figure stable over a period of at least one or six months.

In some cases, the nanoparticle includes a core and/or a coating, wherein the coating preferentially stabilizes the core and/or prevents the core from aggregating and/or leads to the chain arrangement of at least two nanoparticles.

In other cases, the nanoparticle is amorphous and/or crystallized.

In some cases, an iron oxide core is composed of at least one iron atom and at least one oxygen atom.

In some other cases, a core composed of a metal oxide is composed of at least one metal atom and at least one oxygen atom.

In still other cases, a metal core is composed of at least one metal atom.

In one embodiment of the invention, the composition comprises at least one nanoparticle with the cryoprotectant and optionally a liquid such as water.

In another embodiment of the invention, the composition comprises at least one nanoparticle without the cryoprotectant and optionally a liquid such as water.

In another embodiment of the invention, the composition comprises an excipient, an active ingredient, and/or a surfactant.

In another embodiment of the invention, the composition is a preparation or a suspension or a powder, preferably of or with the at least one nanoparticle, optionally with the cryoprotectant.

In one embodiment of the invention, the cryoprotectant is or comprises at least one substance or compound or chemical function or molecule or atom, which protects or maintains or conserves at least one property of the composition, preferably chosen from : i) the chain arrangement or the geometric figure of the at least two nanoparticles, ii) the at least one center for capturing or producing free radicals, iii) the isotonicity or osmolality, preferably of or in the composition, preferably at or below the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, -5, -10, -50, -77, -100, -200 or -273°C.

In one embodiment, the free radical production or capture center is a compound which increases the quantity of free radicals produced or captured when: i) radiation is applied to the center, preferably designated as a radiosensitizer in this case , ii) light is applied to the center, preferably designated as a photosensitizer in this case, iii) thermal radiation or an increase or decrease in temperature is applied to the center, preferentially designated as a thermo-sensitizer in this case. case, iv) a cryogenic treatment or protection is applied to the center, preferably designated as a cryo-sensitizer in this case, v) a magnetic field is applied to the center, preferably designated as a magneto-sensitizer in this case, vi ) ultrasound is applied to the center, preferably designated as a sonosensitizer.

In one embodiment, the center of production or capture of free radicals may be a center of physico-chemical disturbance, such as radiation, with amplification, where the amplification may correspond to an effect of the physico-chemical disturbance. chemical applied to the part of the body which is more important in the presence than in the absence of the center, for example the eradication or death of a tumor could be induced in the presence of the irradiated center while this effect would be absent or less pronounced in the absence of the center.

In one embodiment, the free radical production or capture center is a compound which amplifies or increases the physico-chemical disturbance or the radiation or at least one parameter or property of the physico-chemical disturbance or the radiation or less an effect of radiation or a physicochemical disturbance. When the center is exposed to radiation, light, thermal variation, cryo-treatment or cryoprotection, a magnetic field or ultrasound, it can be designated respectively as a radio-sensitizer, photosensitizer , thermo-sensitizer, cryo-sensitizer, magneto-sensitizer or sonosensitizer.

In another embodiment of the invention, the at least one property of the composition is maintained, or preserved or protected, preferably by the cryoprotectant, when:
At least two nanoparticles remain arranged in a chain or geometric figure,
At least one free radical production or capture center remains included in the nanoparticle or composition or remains active, that is to say capable of producing or capturing free radicals preferentially under the application of a radiation,
and or
The isotonicity, osmolality and/or at least one other property of the nanoparticle or composition does not change or vary by more than 10-3, 1, 5, 10, 25, 50, 75 or 100% , where this percentage is preferably equal to (P2-P1)/P1, preferably measured in absolute terms or values, where P1 and P2 are values of isotonicity, osmolality and/or at least one other property of the nanoparticle or composition at a temperature T1 and T2, respectively,
preferably for or when the temperature of the composition or the nanoparticle is maintained or decreased below or at the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, -5, -10 , -50, -77, -100, -200 or -273 °C.

In some cases, T1 and T2 are two different temperatures.

In some cases, T2 is less than T1, preferably by at least 10-5, 10-3, 10-1, 0, 1, 2, 5, 10 or 103°C.

In some cases, T1 and/or T2 is/are less than 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, -5, -10, -50, -77, -100, - 200 or -273°C.

In certain other cases, T1 and/or T2 is/are greater than -273, -200, -100, -77, -50, -10, -5, 0, 1, 2, 5, 10, 50 , 100 or 103 °C.

In one embodiment of the invention, the cryoprotectant is not used to protect the at least one nanoparticle or the at least one chain against denaturation and/or destruction and/or loss of the primary, secondary, tertiary or quaternary of DNA, RNA, protein(s), lipid(s), enzyme(s) or at least one biological molecule.

In one embodiment of the invention, the cryoprotectant prevents a change, decrease, increase in the size or at least one nanoparticle or chain of more than 100, 50, 10 or 1%, where this percentage is preferably equal to S2-S1/S1 or P2-P1/P1, where S1, P1, S2 and P2 are preferably the sizes and properties of at least one nanoparticle or chain before (S1, P1) and after (S2, P2) the cooling the at least one nanoparticle or chain, preferably by more than 1, 50 or 100°C, preferably from an initial temperature, preferably greater than -273, -100, -50, 0.5 or 10°C, preferably to a final temperature, preferably below 100, 50, 0 or -50°C.

In certain cases, the property of at least one nanoparticle or chain may be: i) the mass percentage of at least one metal in the at least one nanoparticle or chain, ii) the number of nanoparticles in the chain, iii ) the surface charge of the at least one nanoparticle or chain iv) the coercivity or remanent magnetization of the at least one nanoparticle or chain, v) the diameter of the core of the at least one nanoparticle, vi) the thickness of the coating of the at least one nanoparticle, vii) the isotonicity or osmolality of the at least one nanoparticle or chain.

In one embodiment of the invention, the composition can be cooled, preferably below or to 10, 5, 2, 1, 0, -5, -10, -50, -77, -90, -270, or -273°C, or the composition below or at 10, 5, 2, 1, 0, -5, -10, -50, -77, -90, -270, or -273°C, can be achieved or preferably obtained for more than 1 second, 1 minute or 1 hour, preferably without destroying at least one chain or geometric figure, preferably with at least one chain or geometric figure which is maintained or exists. This effect is preferentially due to the presence of the cryoprotectant.

In one embodiment of the invention, the composition is freeze-dried or dried, preferably for more than 1 second, 1 minute or 1 hour, preferably without destroying at least one chain or geometric figure, preferably with at least one chain or geometric figure that is maintained or exists. This effect is preferentially due to the presence of the cryoprotectant.

In one embodiment of the invention, the composition is prepared or used by following at least one of the following steps:

First, the composition is mixed with water to add the cryoprotectant to the at least one chain in liquid suspension.

Next, the composition is freeze-dried to remove water from the composition and preserve the composition in powder form for storage.

Third, the composition is resuspended in water, ready for use and administration to humans.

In one embodiment, the at least one step mentioned above is repeated.

In one embodiment of the invention, the dissociation energy between a chemical group of a molecule of the coating and a chemical group of the core of the nanoparticle is greater than 10 Kcal/mol, 100 KJ/mol or 1 eV/ connection.

In one embodiment of the invention, the dissociation energy between: i) a chemical group of a cryoprotectant molecule and a chemical group of the core of the nanoparticle or ii) a chemical group of a cryoprotectant molecule and a chemical group of the coating of the nanoparticle, is less than 10 Kcal/mol, 100 KJ/mol, or 1 eV/bond.

The invention relates to the composition according to the invention, in which the chemical affinity between the coating and the core of the nanoparticle is greater than: i) the chemical affinity between the cryoprotectant and the coating and/or ii) the affinity chemical between the cryoprotectant and the core of the nanoparticle.

In one embodiment of the invention, the coating is more strongly bound to the core of the nanoparticle or is in stronger interaction with it than the cryoprotectant.

In one embodiment of the invention, the coating cannot be separated or isolated from the nanoparticle core or is inseparable or non-isolatable from the nanoparticle core, preferably using a magnet or magnetic separation or centrifugation.

In another embodiment of the invention, the cryoprotectant may be separated or isolated from the nanoparticle core or is separable or isolable from the nanoparticle core, preferably using a magnet or magnetic separation or centrifugation.

In some cases, this can be evidenced by the fact that: i) the coating and the core of the nanoparticle cannot be separated by a magnet of low intensity, i.e. preferably of lower intensity at 1 T or 1 mT, or a low magnetic field gradient, that is to say preferably a magnetic field gradient less than 1 T or mT per cm preferably of the part of the body, while the cryoprotectant can be preferentially separated from the nanoparticle by such a magnet, ii) the coating and the core of the nanoparticle cannot be preferentially separated by mixing the core and the coating of the nanoparticle in water followed by centrifugation while the cryoprotectant can be preferentially separated from the nanoparticle by being mixed in water with the suspended nanoparticles followed by centrifugation.

In one embodiment, the coating comprises at least one compound or chemical function, which is capable of establishing interactions or weak interactions or weak bonds or covalent bonds with the core of the nanoparticle or at least one chemical function or an atom or ion of the central part of the nanoparticle, in particular iron oxide or any combination or ionic state of iron and/or oxygen. In some cases, such interactions or bonds maintain the coating of the nanoparticle attached or linked or associated with or at the core of the nanoparticle, preferably called bonds or association or attachment or bonding interactions.

In one embodiment of the invention, the nanoparticle consists of a core, a surrounding coating, and association or binding bonds or interactions.

In one embodiment of the invention, the nanoparticle consists of a core and a surrounding coating, which is preferentially associated or linked or attached to the core, preferably in a manner or by sufficiently strong bonds or interactions so that it does not detach or dissociate from the nucleus or remain attached or bound to the nucleus.

In some cases, the chain arrangement or geometric figure of the nanoparticles is maintained or exists or forms in the presence of the coating.

In some cases, in the absence of coating, the chain arrangement or geometric figures do not exist or do not form or exist.

In one embodiment of the invention, the composition comprises a cryoprotectant, which is associated or linked or linked to the nanoparticle, to the core, to the coating, preferably in a manner or by bonds or interactions sufficiently weak to that it detaches or dissociates from the nanoparticle, the core or the coating, preferably when the cryoprotectant is washed away or removed from the nanoparticles by washing with water, by centrifugation or by using a magnet.

In some cases, the cryoprotectant may be dissociated or detached or disassembled or unbound from the nanoparticle, core or coating, preferably such that the chain arrangement or geometry of the nanoparticles is maintained.

In some cases, the cryoprotectant may be associated or attached or assembled or bonded to or with the nanoparticle, core or coating, preferably such that the chain arrangement or geometry of the nanoparticles is maintained, preferably when the composition is cooled, preferably below or to 100, 10, 5, 2, 1, 0, -10, -50, -77, -270, -273 °C, preferably for more than 0.001, 0.1 , 0, 1, 5, 10, 103 or 105 seconds, or when the composition is lyophilized or dried or when the water is removed from the composition partially or completely or when the percentage by mass of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5% or when the composition is in powder form.

In some cases, the chain arrangement or geometric figure of the nanoparticles can be observed by following at least one of the following steps: i) removal of the cryoprotectant from the composition, ii) suspension or resuspension of the composition , preferably lyophilized, preferably in water, iii) depositing a droplet of the composition on a substrate, preferably a carbon grid, iv) waiting for the water to evaporate, and v) observing the arrangement nanoparticles under the electron microscope.

In some cases, preferably, in the absence of coating, chains or geometric figures do not form or exist, while in the presence of coating, chains or geometric figures preferably form or exist .

In certain cases, when the temperature of the at least one nanoparticle is less than or equal to 100, 10, 5, 2, 1, 0, -10, -50, -77, -270 or -273 ° C, preferably during more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the nanoparticle is freeze-dried or dried or when water is removed from the composition or from the at least one nanoparticle partially or completely or when the percentage by mass of water in the composition the at least one nanoparticle is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5%, the chain arrangement or the geometric figure n preferentially does not exist or does not form or is destroyed in the absence of the cryoprotectant, and/or the chain arrangement or the geometric figure preferentially exists or forms or is maintained or is not destroyed in the presence of the cryoprotectant.

In one embodiment of the invention, the coating or at least one compound or chemical function included in the coating is or remains chemisorbed or physisorbed or adsorbed or attached or linked or associated or linked to the core of the nanoparticle, preferably in the presence of the cryoprotectant, surrounding or preferentially coating the nanoparticle, preferably for a duration greater than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, preferentially for or when the composition is in powder form, preferentially for or when the percentage by mass of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5%.

In one embodiment of the invention, the coating or at least one compound or chemical function included in the coating is not or does not remain chemisorbed or physisorbed or adsorbed or attached or bound to or associated with or bound at the heart of the nanoparticle, preferably in the absence of the cryoprotectant, surrounding or preferentially coating the nanoparticle, preferably for a duration greater than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, preferably for or when the composition is in powder form, preferably for or when the percentage by mass of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5%.

In one embodiment, the coating or at least one compound or chemical function included in the coating is linked by bonds or interactions with or to Fe2+ or Fe3+ ions, to OH- hydroxyls, to O2- oxides, to crystalline defects of the core, which can be found in or on the surface of the core of the nanoparticle.

In one embodiment, the coating comprises at least one compound, an atom, an ion or a chemical function such as an acid, carboxylic acid, phosphoric acid or sulfonic acid function, in which the compound, the atom or the ion contained in the coating is capable of establishing interactions or bonds with the nucleus or with at least one atom of the nucleus, a chemical function of the nucleus, a nucleus ion such as Fe2+, Fe3+, Hydroxyl OH-, oxide O2- or a crystal defect in the nucleus.

In one embodiment of the invention, the interactions or bonds exist or are maintained when the composition is cooled, preferably below or to 100, 10, 5, 2, 1, 0, -10, -50, - 77, -270, -273 °C, preferably for more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the composition is freeze-dried or dried or when water is removed from the composition partially or totally or when the percentage by mass of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5% or when the composition is in powder form, preferably in the presence of the cryoprotectant. In certain cases, these interactions or bonds maintain the nanoparticles organized in a chain or in a geometric figure, that is to say that preferably in the absence of these interactions or bonds, the nanoparticles are not organized in a chain or in a geometric figure. .

In some cases, a geometric figure is an assembly or aggregate of nanoparticles forming a geometric figure.

In certain cases, a geometric figure is chosen from the group consisting of: a Balbis, a concave polygon, a constructible polygon, a convex polygon, a cyclic polygon, an equiangular polygon, an equilateral polygon, a Penrose tile, a polyform, a regular polygon, a simple polygon, a tangential polygon, polygons with specific numbers of sides, a henagon, a digon, a triangle, an acute triangle, an equilateral triangle, a heptagonal triangle, an isosceles triangle, an obtuse triangle, Rational triangle, Right triangle, Kepler triangle, Scalene triangle, Quadrilateral, Cyclic quadrilateral, Kite, Parallelogram, Rhombus, Rhomboid, Rectangle, Square, Tangential quadrilateral, Trapezoid, Isosceles trapezoid, Pentagon, Hexagon, Lemoine hexagon , Heptagon, Octagon, Nonagon, Decagon, Hendecagon, Dodecagon, Tridecagon, Tetradecagon, Pentadecagon, Hexadecagon, Heptadecagon, Octadecagon, Enneadecagon, Icosagon, Swastika, Star polygon, Pentagram - star polygon, Hexagram, Star of David, Heptagram, Octagram, canvas of Lakshmi, Decagram - star polygon, Annulus, Arbelos, Circle, Archimedes' twin circles, Bankoff's circle, Circumference, Disc, Incircumference and excircumference of a triangle, Nine-point circle, Circular sector, Circular segment, Crescent, Indalo , Lens, Moon, Reuleaux polygon, Reuleaux triangle, Salinon, Semicircle, Tomahawk, Triquetra, Heart, Archimedean spiral, Astroid, Cardioid, Deltoid, Ellipse, Heart, Heartagon, Various lemniscates, Oval, Cartesian oval, Cassini Oval, Booth Oval, Ovid, Superellipse, Taijitu, Tomoe, and/or Magatama.

In one embodiment of the invention, the nanoparticle, the suspension, the composition or the assembly of nanoparticles is stable, preferably for a period of time, preferably its duration of stability, which is greater than 10-10, 5, 10, 1050 or 10100 minute(s), 1, 2, 3, 4, 5, 6, 10, 12, 24 or 36 months.

In some cases, the nanoparticles, nanoparticle suspension, composition or assembly of nanoparticles may be stable at a nanoparticle concentration greater than 1, 5, 10, 50, 100, 200, 500 or 1000 mg of nanoparticles per mL solvent, water, matrix or part of the body surrounding or comprising or incorporating the nanoparticles or the composition.

In one embodiment of the invention, the body part is the body part of an individual, animal or human.

In one embodiment of the invention, the body part comprises more than or at least 1, 2, 5, 10 or 100 organism(s), apparatus(es), organ(s), tissue(s), cell Similar or different biomolecule(s) or biomolecule(s).

In some cases, the body part may be all or part of the head, neck, shoulder, arm, leg, knee, foot, hand, ankle, elbow, trunk, lower limbs or upper limbs.

In certain other cases, the body part may be or belong to an organ, musculoskeletal, muscular, digestive, respiratory, urinary, female reproductive, male reproductive, circulatory, cardiovascular, endocrine, circulatory, lymphatic, nervous (peripheral) system or not), ventricular, enteric, sensory, or integumentary nervous, to a reproductive organ (internal or external), to a sensory organ, to endocrine glands. The organ or body part may be the human skeleton, joints, ligaments, tendons, mouth, teeth, tongue, salivary glands, parotid glands, submandibular glands, sublingual glands, pharynx , esophagus, stomach, small intestine, duodenum, jejunum, ileum, large intestine, liver, gallbladder, mesentery, pancreas, nasal cavity, pharynx, larynx, trachea , bronchi, lungs, diaphragm, kidneys, ureters, bladder, urethra, ovaries, fallopian tubes, uterus, vagina, vulva, clitoris, placenta, testes, epididymis, vas deferens, seminal vesicles, prostate, bulbourethral glands, penis, scrotum, pituitary gland, pineal gland, thyroid gland, parathyroid glands, adrenal glands, pancreas, heart, arteries, veins, capillaries, lymphatic vessels, lymph nodes, bone marrow, thymus, spleen , the lymphoid tissue associated with the intestine, the tonsils, the brain, the cerebrum, the cerebral hemispheres, the diencephalon, the brainstem, the midbrain, the pons, the medulla oblongata, the cerebellum, the spinal cord, choroid plexus, nerves, cranial nerves, spinal nerves, ganglia, eye, cornea, iris, ciliary body, lens, retina, ear, outer ear, earlobe, eardrum, middle ear, ossicles, inner ear, cochlea, vestibule of the ear ear, semicircular canals, olfactory epithelium, tongue, taste buds, mammary glands or skin.

In some cases, the body part or organ may belong to the bloodstream or circulatory system.

In some cases, the body part may be or include at least one tumor, cancer, virus, bacteria or pathological cell.

In one embodiment of the invention, the body part is or includes water, an excipient, a solution, a suspension, at least one chemical element, an organic material or a gel, which may be synthetic or produced by a living organism.

Preferably, the body part of an individual, also referred to as the body part, represents or is part of an individual or an entire individual, where the individual is preferably a human, an animal, or a organism, preferably a living or inactivated or dead organism, comprising at least one prokaryotic or eukaryotic cell.

In one embodiment of the invention, the part of the body is living (or not), is any tissue, water, medium, substance, cell, organelle, organ protein, lipid, DNA, RNA, biological material, preferably located in a specific region of an individual, preferably originating or extracted from this region.

In one embodiment of the invention, the body part comprises a pathological site, a healthy site, and/or a region of nanoparticles or composition.

In one embodiment of the invention, the body part is or includes a pathological site or pathological cells.

In some cases, the pathological site may be defined as an unhealthy site, or a site that is in a different state from that of a healthy individual, or from the site of an unhealthy individual. It may include pathological cells, such as tumor cells, bacteria, eukaryotic or prokaryotic cells, as well as viruses or any other pathological material. Pathological cells may be cells that: i) are not arranged or function as they usually do in a healthy individual, ii) divide more rapidly than healthy cells, iii) are healthy cells that have undergone transformation or modification, iv) are dead, sometimes due to the presence of a virus or other organisms, or v) are in contact, interacting, with foreign material not belonging to the individual, such as viruses, where viruses can eventually enter, colonize or replicate in these cells. In some cases, pathological cells may be likened to viruses or other organisms or entities that colonize cells or target cells or destroy cells or use cells or interact with cells, preferably to enable their own reproduction, multiplication, survival or death. In some cases, a pathological site may include healthy cells, which are preferably lower in number, activity or proliferation than pathological cells.

In one embodiment of the invention, the body part is or includes a healthy site or healthy cells. In some cases, the healthy site may be defined as a site or region that includes one or more healthy cells, where a healthy cell may be defined as a cell that belongs to a healthy individual or to the body part of a healthy individual. .

In certain cases, the healthy site can surround the pathological site when it is preferentially located at a distance less than 1 or 10-9 m from the pathological site.

In some cases, the composition or body part may be exposed to radiation.

Preferably, the radiation is chosen from the group consisting of: i) a magnetic, electric or electromagnetic field or wave, a particle wave or radiation, ii) laser light, iii) light produced by a lamp, iv) light emitted at a single wavelength, v) light emitted at several wavelengths, vi) ionizing radiation, vii) microwave, viii) radio frequencies, and ix) sound, ultrasound, an infrasound or acoustic wave.

In one embodiment of the invention, the radiation has at least one of the following properties:
i) it has a power or power density of less than 1000, 10 or 1 W (Watt), preferably per cm, W per cm2, or W per cm3, preferably of the body part or composition, or preferably per gram or milligram, preferably per body part or composition,
ii1) it has an energy or energy density less than 105, 103, 100 or 1 W.sec per cm, W.sec per cm2, or W.sec per cm3, preferably of the body part or composition,
ii2) it has an energy or energy density less than 105, 103, 100 or 1 W.sec per gram or milligram, preferably of body part or composition,
ii3) it has an energy or energy density less than 105, 103, 100 or 1 (Joule), J, preferably per cm, per cm2, or per cm3, preferably of the body part or composition , preferably per gram or milligram, preferably of the composition or body part,
(iii) it has a frequency less than 105 100, 10, 1, 10-1 or 10-3 MHz,
iv) it has a penetration depth in the body part less than 1010, 105, 103, 10, 1, 0 or 10-5 cm,
v) it has a wavelength less than 1010, 105, 103, 100, 50, 10, 10, 5, 2, 1, 0.1 or 0 nm.

In another embodiment of the invention, the radiation has at least one of the following properties:
i) it has a power or power density greater than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 W (Watt), preferably by cm, W per cm2, or W per cm3, preferably per body part or composition, or preferably per gram or milligram, preferably per body part or composition,
ii1) it has an energy or energy density greater than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 W.sec per cm, W .sec per cm2, or W.sec per cm3, preferably of the body part or composition,
ii2) it has an energy or energy density greater than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 W.sec per gram or milligram , preferably part of the body or composition,
ii3) it has an energy or energy density greater than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 (Joule), J, of preferably per cm, per cm2, or per cm3, preferably of the body part or the composition, preferably per gram or milligram, preferably of the composition or the body part,
iii) it has a frequency lower than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 MHz,
(iv) it has a penetration depth in the body part of less than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 cm,
v) it has a wavelength less than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 103 or 105 nm.

In one embodiment of the invention, the nanoparticle or composition is administered to or into the body part.

In another embodiment of the invention, the nanoparticle or composition is administered at a distance of less than 1 or 10-9 m from the body part.

In yet another embodiment of the invention, the nanoparticle or composition is administered at a distance of more than 1 or 10-9 m from the body part.

In another embodiment of the invention, the nanoparticle or the composition is administered to or in the part of the body following at least one of the following routes of administration: local, enteral, gastrointestinal, parenteral, topical, oral, inhalation, intramuscular, subcutaneous, intratumoral, in an organ, in a vein, in arteries, in the blood or in a tissue.

In some cases, the nanoparticles, nanoparticle suspension, composition or assembly of nanoparticles may be stable when at least one nanoparticle is not degraded or does not partially or completely lose its coating or can be administered to a part of the body or preserves or maintains its chain arrangement or geometric figure.

In other cases, the nanoparticles, the nanoparticle suspension, the composition or the nanoparticle assembly may be stable when the optical density of the nanoparticles, the nanoparticle suspension, the composition or the nanoparticle assembly, preferably mixed in water, preferably measured at 480 nm or another fixed wavelength, does not decrease by more than 1, 5, 10, 50, 75 or 90% or by more than 10-10, 10-3 , 10-1, 0.5 or 0.7, preferably within 1, 5, 10, 10, 10 or 10 seconds following homogenization or mixing or optical density measurement or absorption measurement of this suspension or composition.5 or 0.7, preferably within 1, 5, 10, 103, 107 or 1020 seconds following homogenization or mixing or measurement of the optical density or measurement of the absorption of this suspension or composition. This percentage may be equal to (ODB -ODA)/ODB or ODA/ODB, where ODB is the optical density of the nanoparticles, the suspension of nanoparticles, the composition, or the assembly of nanoparticles measured before homogenization or the mixing or optical density measurement or absorption measurement of the nanoparticle, suspension, composition, or assembly of nanoparticles and ODA is the optical density of the nanoparticles, nanoparticle suspension, composition or the assembly of nanoparticles measured after homogenization or mixing or the measurement of the optical density or the measurement of the absorption of the nanoparticles, the suspension of nanoparticles, the composition or the assembly of nanoparticles.

In some cases, the composition may be stable or considered stable when measured stable at a certain first time t0 and at a certain second time t1, where t1 follows t0 or is separated from t0 by a period of timeΔT of at least 1 second, 1 minute, 1 hour, 1 day, 1 month, 3 months, 6 months, 1, 2, 5 or 10 years. DuringΔ T, the composition is preferably stored.

In some cases, the nanoparticle can be suspended in a liquid or dispersed in a matrix or part of the body to give a homogeneous dispersion of nanoparticles or a very stable composition or suspension of nanoparticles.

In one embodiment of the invention, the cryoprotectant serves to keep the chain or geometric figure stable for a period of time, preferably Δ T, preferably at least one or six months. In this case, the chain or the geometric figure can preferentially be observed or measured, preferably by electron microscopy, at a first moment t0 and at a second moment t1, where t1 is preferentially separated from t0 byΔ T.

In one embodiment of the invention, the cryoprotectant protects or maintains the arrangement of at least two nanoparticles in at least one chain or in at least one geometric figure.

In certain cases, the protection or maintenance of the arrangement of at least two nanoparticles in at least one chain or in at least one geometric figure is done at a temperature preferably less than or equal to 103, 500, 100, 50, 10 , 5, 2, 1, 0, -5, -10, -20, -50 -100, -200, or -273 °C of the composition.

In other cases, the protection or maintenance of the arrangement of at least two nanoparticles in at least one chain or in at least one geometric figure is done at a temperature of the composition preferably greater than or equal to -273, - 200, -100, -50 -20, -10, -5, 0, 1, 2, 5, 10, 50, 100, 500, or 103°C.

In certain cases, the protection or maintenance of the arrangement of at least two nanoparticles in at least one chain or in at least one geometric figure occurs for a period preferably greater than 10-10, 10-5, 10-3 , 10-1, 0, 1, 2, 5, 10, 102, 103 or 105 minutes or seconds.

In certain other cases, the protection or maintenance of the arrangement of at least two nanoparticles in at least one chain or in at least one geometric figure occurs for a duration preferably less than 1010, 105, 103, 10, 5, 2, 1, 0, 10-1, 10-3 or 10-5 minutes or seconds.

The invention relates to the composition according to the invention, where the at least one chain is in liquid suspension and the composition preferably presents at least one of the following properties:
i) it is isotonic to animal or human plasma or blood,
ii) the volume occupied by the water in the composition is greater than the volume occupied by the at least one chain in the composition, and
iii) the percentage by mass of water in the composition is greater than the percentage by mass of the at least one chain in the composition.

In one embodiment of the invention, the at least one chain is in liquid suspension when mixed with a liquid. In some cases, the liquid may be water, an isotonic liquid, a liquid comprising an excipient, a surfactant, or an oil.

In certain cases, the composition is isotonic, that is to say it preferentially presents:
the same osmotic pressure as that of a part of the body such as a cell, a body fluid, plasma, blood, preferably of a human being or an animal,
the value of abs(OPBP - OPComp )/OPBP is less than 100, 50, 10, 5, 2, 1, or 10-3%, where OPBP and OPComp are the osmotic pressures of the body part and composition, respectively .

In another embodiment of the invention, OPBP and/or OPComp is/are greater than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 50 or 100 bar or atm.

In another embodiment of the invention, OPBP and/or OPComp is/are less than 1010, 105, 103, 10, 0, 10-1, 10-3, 10-5, or 10-10 bar or atm.

In one embodiment, the value of abs(OPBP - OPComp )/OPBP is kept low or the composition is kept isotonic to avoid an increase in blood pressure, preferably during or after administration of the composition to the part of the body.

In some cases, the composition may be hypertonic, that is, preferably in this case, the composition causes shrinkage of at least one cell or part of the body.

In other cases, the composition may be hypotonic, that is, preferably in this case, the composition causes at least one cell or part of the body to swell.

In still other cases, the composition may be isotonic, that is, preferably in this case the composition produces no change in the volume of the cells or body parts or a lesser change in the volume of the cells or parts of the body. cells or parts of the body than for a hypertonic or hypotonic composition.

In one embodiment of the invention, the solutes or solute compounds in the composition, preferably the nanoparticle, the chain of nanoparticles and/or the cryoprotectant, have a concentration, preferably designated by Csolute, preferably designated by Csolute, outside a cell or outside a part of the body, preferably designated by Csolute, inside a cell or inside a part of the body, which is such that abs(Csolute,outside - Csolute,inside) / Csolute,outside is less than or equal to 100, 50, 25, 10, 5, 2, 1, 0, 10-1, 10 -3 or 10-5%.

In some other cases, (Csolute,outside - Csolute,inside) / Csolute,outside is greater than or equal to 10-5, 10-3, 10-1, 0, 1, 2, 5, 10, 25, 50, 75 or 99%.

In one embodiment of the invention, the composition is isosmotic or the osmolarity or osmotic pressure of the composition, preferably outside at least one cell or part of the body is the same as the osmolarity or the osmotic pressure of an intracellular medium or of a part of the body or of a composition included inside at least one cell or part of the body.

In some cases, body part may refer to a fluid or liquid medium included in a body part, such as the intracellular medium or plasma or blood.

In some cases, the composition is included in the body part, preferably after or with administration to the body part.

In some other cases, the composition is included external to the body part, preferably before or without administration to the body part.

In some cases, the composition may be hypo-osmotic.

In some other cases, the composition may be hyperosmotic.

In one embodiment of the invention, the osmolality or osmolarity of the composition is greater than or equal to 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10, 50 , 100, 200, 250, 290, 300, 310, 350, 400, 500, 103, 105 or 1010 mOsm/kg or mOsm/L.

In another embodiment of the invention, the osmolality or osmolarity of the composition is less than or equal to 1010, 105, 103, 500, 350, 310, 300, 250, 200, 100, 10, 5, 1, 0, 10-3, 10-5 or 10-10 mOsm/kg or mOsm/L.

In another embodiment of the invention, the osmolality of the composition is equal to, close to, or does not differ by more than 1, 5, 10, 50 or 100% from the osmolality of human plasma, preferably between 275 and 299 milli-osmoles per kilogram.

In another embodiment of the invention, the hydrostatic pressure or the osmotic pressure or the arterial pressure or the systolic pressure or the diastolic pressure or the pressure of the composition or in the presence of the composition or following the administration of the composition or applied by the composition to the part of the body or the force pushing the composition or at least one compound or solute of the composition preferably per unit area of the part of the body, preferably in some cases from outside the part of the body or at least one cell of the body part towards the interior of the body part or towards the interior of at least one cell of the body part, preferably in other cases of the interior of the body part or from the interior of at least one cell of the body part to the exterior of the body part or to the exterior of at least one cell of the body part, is designated byξ.

In some cases, ξ may be less than or equal to 105, 103, 500, 200, 180, 150, 140, 100, 50, 20, 10, 1, 0, 10-1, 10-3 or 10-5 mmHg or PSI or bar or mbar.

In certain other cases, ξ is greater than or equal to 10-5, 10-3, 10-1, 0, 1, 5, 10, 100, 120, 150, 500, 103, 105, or 1010 mmHg or PSI or bar or mbar.

In one embodiment of the invention, the volume occupied by the water or a liquid and preferably the cryoprotectant in the composition is greater than the volume occupied by the at least one chain in the composition. In this case, the at least one chain is preferably suspended in a liquid, preferably water, which comprises the cryoprotectant. In this case, the volume of liquid is preferably kept sufficiently large or larger than the volume occupied by the at least one chain in the composition to allow the suspension of the at least one chain in the composition or the movement or Brownian movement of at least one string in the composition.

In certain cases, the volume occupied by the water or the liquid and preferably the cryoprotectant in the composition is greater than the volume occupied by the at least one chain in the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

In certain other cases, the volume occupied by the water or the liquid and preferably the cryoprotectant in the composition is less than the volume occupied by the at least one chain in the composition, by an α factor of at least 1.1, 2 , 5, 10 or 103 . In this case, the composition is preferably in powder form.

In certain cases, α can be equal to Vliq /Vchain, where Vliq and Vchain are the volumes occupied by the liquid preferentially comprising the cryoprotectant and at least one chain in the composition, respectively, where the two volumes can preferentially be measured by separating the at least one chain from the liquid, for example using magnetic separation or using a magnet to attract the chains and separate them from the liquid.

In another embodiment of the invention, the mass of water or liquid and preferably of cryoprotectant in the composition is greater than the mass of at least one chain in the composition. In this case, the at least one chain is preferably suspended in a liquid, preferably water, which comprises the cryoprotectant. In this case, the mass of liquid is preferably kept sufficiently large or greater than the mass of the at least one chain in the composition to allow the suspension of the at least one chain in the composition or the movement or Brownian movement of the at least one string in the composition.

In certain cases, the mass of water or liquid and preferably of cryoprotectant in the composition is greater than the mass of the at least one chain in the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

In certain other cases, the mass of water or liquid and preferably of cryoprotectant in the composition is less than the mass of the at least one chain in the composition, by a factor α of at least 1.1, 2, 5 , 10 or 103 . In this case, the composition is preferably in powder form.

In some cases, α may be equal to mliq /mchain, where mliq and mchain are the masses of the liquid preferably comprising the cryoprotectant and at least one chain in the composition, respectively, where the two masses may be preferentially measured by separating the at least a chain of the liquid, for example using magnetic separation or using a magnet to attract the chains and separate them from the liquid, where mchain and/or mliq can be measured preferably by or after evaporation of the liquid from the composition or lyophilization or drying of the composition.

In one embodiment of the invention, the percentage by mass of water or liquid, preferably comprising the cryoprotectant, preferably not comprising the at least one chain, in the composition, is greater than the percentage by mass of the at least a string in the composition.

In certain cases, the percentage by mass of water or liquid preferably comprising the cryoprotectant, preferably not comprising the at least one chain, in the composition is greater than the mass of the at least one chain in the composition, by one α factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

In one embodiment of the invention, the percentage by mass of water or liquid, preferably comprising the cryoprotectant, preferably not comprising the at least one chain, in the composition, is less than the percentage by mass of the at least a chain in the composition by an α factor of at least 1, 1, 2, 5, 10 or 103. In this case, the composition is preferably in powder form.

In some cases, α may be equal to Pliq /Pchain, where Pliq and Pchain are the mass percentages of the liquid preferably comprising the cryoprotectant and at least one chain in the composition, respectively, where both mass percentages may be preferentially measured by separating the at least one chain from the liquid, for example using magnetic separation or using a magnet to attract the chains and separate them from the liquid, where Pchain and/or Pliq can be measured preferably by or after evaporation of the liquid from the composition or lyophilization or drying of the composition.

The invention also relates to the composition according to the invention, where the at least one chain is in powder form, and the composition preferably presents at least one of the following properties:
i) the volume occupied by the water or liquid and preferably by the cryoprotectant in the composition is less than the volume occupied by the at least one chain in the composition, and
ii) the percentage by mass of water or liquid and preferably of cryoprotectant in the composition is less than the percentage by mass of at least one chain in the composition,
wherein the composition is preferably lyophilized, desiccated, dried or dehydrated.

In some cases, the composition is freeze-dried, desiccated, dried or dehydrated when water or liquid is removed, preferably partially or completely, from the composition.

The invention also relates to the composition according to the composition, wherein the composition is lyophilized or desiccated or dried or dehydrated or treated by lyophilization or desiccation or removal of water or removal of liquid or dehydration.

In one embodiment of the invention, the composition is dehydrated or water or liquid is removed or absent from the composition. In this case, the percentage by mass of water or liquid in the composition is preferably greater before dehydration of the composition or before elimination of water or liquid from the composition than after dehydration of the composition or after removal of water or liquid from the composition. In this case, the volume of water or liquid occupied in the composition is preferably greater before the composition is dehydrated or before the water or liquid is removed from the composition than after the composition is dehydrated or after until water or liquid is removed from the composition. In this case, the water or liquid is removed from the composition without removing the at least one chain and preferably also the cryoprotectant from the composition.

In one embodiment of the invention, the percentage by weight of water in the composition is greater than 10-3, 10-1, 0, 1, 5, 10, 25, 50, 80 or 99%, preferably before water is or has been removed from the composition.

In another embodiment of the invention, the percentage by weight of water in the composition is less than or equal to 100, 99, 80, 50, 25, 10, 5, 2, 1 or 0%, preferably after whether water is or has been removed from the composition.

In another embodiment of the invention, the composition is freeze-dried or the composition is first cooled, preferably below or at 103, 100, 50, 10, 0, -10, -50, -77, -200 , -273°C, and the water or liquid is then removed from the composition, preferably without removing the at least one chain and/or the cryoprotectant from the composition.

In another embodiment of the invention, the composition is lyophilized, dehydrated, dried and/or desiccated, and the water or liquid is removed from the composition preferably without removing the at least one chain and/or the cryoprotectant. of the composition.

In one embodiment, the composition is included in a tablet or is in the form of a tablet or powder, preferably after removal of water from the composition.

In one embodiment, the composition is stored or retained or unused, preferably for more than 0, 1, 10, 102, 103, 105, or 1010 hour(s), day(s), month, year(s) , preferably without being used or administered to any part of the body, preferably in powder form, preferably without losing at least one of its properties or without losing its nanoparticle coating or its nanoparticle core, partially or totally.

In one embodiment of the invention, the humidity or water content of the composition is less than or equal to 100, 75, 50, 25, 10, 5, 4, 2, 1 or 0% w/w, preferably in weight of water for the weight of the composition or in weight of water for the weight of the composition or in mass of water for the mass of the composition or in mass of water for the mass of the composition, preferably after removal of water from the composition.

In another embodiment of the invention, the humidity or water content of the composition is greater than or equal to 0, 1, 2, 4, 5, 10, 25, 50, 75 or 100% w/w , preferably in weight of water for the weight of the composition or in weight of water for the weight of the composition or in mass of water for the mass of the composition or in mass of water for the mass of the composition , preferably before the water has been removed from the composition.

The invention also relates to the composition according to the invention, where the at least one chain and preferably the cryoprotectant, which is/are preferably in powder form, is/are suspended or resuspended in a liquid or water or treated by suspension or resuspension in a liquid or water, preferably more than 1, 2, 5 or 10 times. In this case, the water is preferentially eliminated from the composition before it is suspended or resuspended in a liquid or water.

The invention also relates to the composition according to the invention, in which the coating is chosen from the group of compounds consisting of: 1) citric acid, 2) oleic acid, 3) polymethacrylic acid, 4) poly(ethylene oxide)-b-poly(methacrylic acid), 5) polyacrylic acid (PAA), 6) polylactic acid, 7) poly(ethylene oxide)-block-poly(glutamic acid ), 8) phosphonic acid 9) albumin, 10) alendronate, 11) alginate, 12) Au, Al2 O3, 13) aluminum, 14) Aluminum hydroxide, 15) Arabinogalactan, 16) Bentonite, 17) Carboxymethylcellulose, 18) Cellulose, 19) Chitosan, 20) Cholesterol, 21) Citrate, 22) Dextran, 23) Dimercaptosuccinic acid, 24) Dopamine, 25) DOPC or Dioleoylphosphatidylcholine or phospholipd, 26) DTAP or Di (tert.-amyl)peroxide, 27) DVB or Divinylbenzene, 28) Ethylcellulose, 29) Erythrocyte, 30) at least one fatty acid, 31) Ferrite, 32) Folic acid, 33) Gelatin, 34) Serum albumin, preferably human, 35) Liposome, 36) MIPS or Inositol-3-phosphate synthase, 37) MnO or manganese oxide, 38) Mn3 O4, 39) Oleic acid, 40) at least one polymer or enantiomer, 41) PEI or Polyetherimide, 42) PEG or polyethylene glycol, 43) poly(ethylene oxide) or PEO, 44) PGA or polyglycolic acid, 45) PLA or poly(lactic acid), 46) PLGA or PLG or poly(lactic-co-glycolic acid ), 47) phosphatidylcholine, 48) Phosphorylcholine, 49) Pluronic, 50) Polyacrylamide, 51) Polyacrylic acid or PAA, 52) Polyaniline, 53) Polyethylene glycol with/without terminal carboxyl groups, 54) Peptide or Polypeptide, 55) Poly( vinyl alcohol) or PVA, 56) Ploy(N-isopropylacrylamide) or PIA, 57) Poly(vinylpyrrolidone) or PVP, 58) Poly(oligoethylene oxide) or POO, 59) Poly(N,N-dimethyl ethylamino acrylate, 60) Poly(imine), 61) Poly(acrylic acid), 62) Poly-D-L lactide, 63) Polyalkylcyanoacrylate, 64) Polymer such as PAMAM or Poly(amidoamine) or PDMAEMA or poly(2-(dimethylamino)ethyl methacrylate) or PPEGMA or Poly (ethylene glycol) methyl ether methacrylate, 65) Poly NIPAAM or Poly (N-isopropylacrylamide) or temperature sensitive polymer or temperature sensitive polymer 66) Polyacrylic acid, 67) Polydipyrrole or dicarbazole, 68) Poly- L-lysine, 69) Polymethylmethacrylate, 70) Polymersome, 71) Polystyrene, 72) PVA or polyvinyl alcohol, 73) PVP or polyvinylpyrrolidone, 74) Silica, preferably amorphous or mesoporous, 75) Silane, 76) SiO2, 77) Oleate sodium, 78) Starch, 79) Styrene, preferably styrene-divinylbenzene, 80) TaOx, 81) ZrO2, 82) at least one metal or a semi-metal, 83) at least one metal oxide or a semi-metallic oxide , 84) at least one alkali metal, 85) at least one alkaline earth metal, 86) at least one transition metal, 87) at least one post-transition metal, 88) at least one metalloid, 89) at least a lanthanide, 90) at least one actinide, 91) at least one non-metal, 92) at least one halogen, 93) at least one noble gas, and 94) any derivative or combination of any of these compounds.

The invention also relates to the composition according to the invention, in which the coating is chosen from the group of the family of compounds consisting of: i) polysaccharides such as agarose, alginate, carregeenan, chitosan, dextran, haparin, gum arabic, pullulan and starch, ii) acids such as citric acids, oleic acids, polymethacrylic acid, poly(ethylene oxide)-b-poly(methacrylic acid, polyacrylic acid or PAA, polylactic acid, poly(ethylene oxide)-blockpoly(glutamic acid, phosphonic acid, dimercaptosuccinic acid, fatty acid, folic acid, oleic acid, poly(lactide) acid or PLA, PAA or polyacrylic acid, compounds comprising at least one carboxylic function, iii) Polymers such as Dextran, Poly(ethylene oxide), Poly(vinyl alcohol), Ploy(N-isopropylacrylamide), Poly(vinylpyrrolidone), Poly(oligoethylene oxide), Poly(N, N-dimethyl ethylamino acrylate), Poly(imine), Poly(acrylic acid), iv) carboxylates, v) inorganic compounds such as SiO2, Al2 O3, ZrO2, ferrites, MnO, Mn3 O4, Au, Bentonite, carbon such as inactivated, activated, graphitized carbon, vi) at least one metal, vii) organic compounds such as MIP, Cellulose, DV8, Ppy, Chitosan, Polyacrylamide, alginate, PEI, surfactants, viii) compounds comprising Phosphate , ix) compounds comprising Silica, x) compounds comprising Gold, xi) compounds comprising a Dextran base, xii) compounds comprising PEG, xiii) compounds comprising PVA, xiv) compounds comprising Alginate , xv) compounds comprising Chitosan, xvi) compounds comprising the chemical function Alcohol, xvii) compounds comprising the chemical function Amide, xviii) compounds comprising the chemical function Aldehyde.

The invention also relates to the composition according to the invention, in which the coating has at least one chemical group chosen from the group of i) OH-, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acidic derivative of at least one of these compounds.

The invention also relates to the composition according to the invention, in which the coating or the coating material comprises a chemical function, preferably chosen from the group consisting of i) OH-, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acid derivative of at least one of these functions, which has an interaction or forms a chemical bond with an atom or a chemical group located on the surface of the nucleus, preferably a hydroxyl group (Fe- OH), where the interaction or bond is preferentially chosen from the group consisting of: i) an electrostatic interaction or bond, i.e. preferentially due to the difference in charge between the coating and the surface or the core of the nucleus, ii) a hydrophobic interaction or bond, iii) a chelating interaction or bond, iv) a metallic interaction or bond, v) a covalent interaction or bond.

In one embodiment of the invention, the cryoprotectant interacts or forms hydrogen or van der walls or London bonds or interactions with the core or coating of the nanoparticle, preferably when the water molecules are displaced. or cooled.

In one embodiment of the invention, as the cryoprotectant replaces the water molecules, preferably when the composition is cooled, the coating and/or the core of the nanoparticle preferentially retains its structure and function and/or at least two nanoparticles remain arranged in a chain.

In one embodiment, the composition comprising the cryoprotectant does not comprise ice, while the composition not comprising the cryoprotectant comprises ice or the composition comprising the cryoprotectant comprises more ice than the composition not comprising the cryoprotectant, preferably when the composition is cooled and/or is freeze-dried,

In one embodiment of the invention, the composition comprising the cryoprotectant has a melting point lower than that of the composition not comprising the cryoprotectant.

In one embodiment of the invention, the cryoprotectant is a non-permeable cryoprotectant.

In certain cases, the cryoprotectant may be chosen from: a sugar, trehalose, sucrose, starch, hydroxyethyl starch, polyvinylpyrrolidone and/or polyethylene oxide.

In some cases, the cryoprotectant may be a non-permeable cryoprotectant, that is, it preferentially does not penetrate cells or coatings and therefore preferentially remains extracellular or outside the coating when the composition is cooled. .

In some cases, the composition is cooled during or for cryopreservation of the composition.

In some cases, a non-permeable cryoprotectant is used to protect the cells while the composition cools.

Preferably, when cooling or slow cooling or slow freezing is applied to the composition or the composition is cooled, preferably slowly, there is sufficient time for the water or liquid included in the nanoparticle, preferably in the coating of the nanoparticle, exits from the coating, preferably under the effect of osmotic pressure, which preferably results in a reduction in the volume of the coating. In the absence of a cryoprotectant, such behavior may destroy or damage the coating and/or result in destruction of the nanoparticle chain arrangement.

In one embodiment, the cryoprotectant is permeable. Preferably, a permeable cryoprotectant can penetrate a cell or coating and preferably preserve the cell or coating, preferably from osmotic pressure or damage or destruction.

In some cases, the cryoprotectant may be dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, or propylene glycol.

In some cases, the cryoprotectant can be used to induce vitrification of the intracellular environment or the coating or the internal part or the surface part of the coating or the nanoparticle or the core, preferably before the formation of ice crystals , preferably in the composition, preferably by preventing excessive volume loss of the cell or coating or nanoparticle or core.

In one embodiment of the invention, the cryoprotectant is chosen from the group consisting of: i) a natural product, ii) a product based on soy flour, iii) a carbohydrate, iv) a lipid, v) a saccharide, vi) a zwitterionic molecule, vii) l-carnitine, and viii) antifreeze compounds such as antifreeze proteins.

The invention also relates to the composition according to the invention, in which the coating or at least one of its functions or chemical atoms forms a first type of interaction or chemical bond with an atom or a chemical function of the nucleus, in which the cryoprotectant or at least one of its functions or chemical atoms forms a second type of interaction or chemical bond with the coating or at least one of its functions or chemical atoms,
where the first type of interaction or chemical bond is preferentially different from the second type of interaction or chemical bond,
in which the first type of interaction or chemical bond is preferentially chosen from the group consisting of: i) an electrostatic interaction or bond, that is to say preferentially due to the difference in charge between the coating and the surface or the core, ii) a hydrophobic interaction or bond, iii) a chelating interaction or bond, iv) a metallic interaction or bond, and v) a covalent interaction or bond,
wherein the second type of chemical interaction or bond is preferably a hydrogen, London or Van Der Walls interaction or bond.

In certain cases, the coating or the coating material may comprise a chemical function, preferably chosen from the group consisting of i) OH-, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic derivative or acid of at least one of these functions.

In certain other cases, the core may comprise a chemical function, preferably located on the surface of the core of the nanoparticle, preferably a hydroxyl group (Fe-OH).

The invention also relates to the composition according to the composition, in which the cryoprotectant is chosen from the group consisting of: 1) Acetamide, 2) Acetate, 3) Albumin, 4) Amino acids, 5) Ammonium acetate, 6) Arginine , 7) Alcohols containing at least one or two hydroxyl groups, 8) Bridger, 9) Choline, magnesium chloride and sodium bromide, 10) Diethyl glycol, 11) Dimethylacetamide, 12) Dimethyl sulfoxide (DMSO), 13) Disaccharide, 14) Erythritol, 15) Ethanol, 16) Ethylene glycol, 17) Formamide, 18) Fructose, 19) Glucose, 20) Glycerol, 21) Glycerol 3-phosphate, 22) Glycol such as Diethyl glycol or Triethylene glycol, 23) Glycine, 24) Lactose, 25) L-tyrosine, 26) lysine hydrochloride, 27) Mannitol, 28) MDP (2-Methyl-2,4-pentanediol), 29) Phenylalanine, 30) Planic, 31) Polymers, 32) Polyethylene glycol tel as PEG4000, polyethylene glycol succinate, distearoylphosphatidyl ethanolamine-folate-modified polyethylene glycol, 33) Polyethyleneimine (PEI), 34) Polyvinylpyrrolidone (PVP), 35) Proline, 36) Propylene glycol, 37) Protein, 38) Pyridine (Pyridine -N-Oxide), 39) Ribose, 40) Sarcosine, 41) Serine, 42) Serum albumin, 43) Sodium bromide, 44) Sodium chloride, 45) Sodium dodecyl sulfonate, 46) Sodium glutamate, 47) Iodide sodium, 48) Sodium sulfate, 49) Sorbitol, 50) Starch (hydroxyethyl starch), 51) Sugar, 52) Sucrose, 53) Cell bank series, 54) Trehalose, 55) Triethylene glycol, 56) Trimethylamine, 57) Tween 80, 58) Tryptophan, 59) Valine, 60) Xylose, and 61) a combination or derivative of any of these compounds.

In one embodiment of the invention, the composition is dried or freeze-dried or dehydrated or dried. In this case, the percentage by mass of water or liquid in the composition is preferably less than 100, 75, 50, 75, 20, 10, 5, 2, 1, 0, 10-1, 10-3 or 10 -5%. In this case, the quantity of water or liquid in the composition is preferably less than 100, 50, 10, 5, 2, 1, 10-1, 10-3 or 10-5 grams of water or liquid in the composition per gram of composition.

In certain other cases, the composition may be undried, undehydrated, or undehydrated or dried. In this case, the percentage by mass of water or liquid in the composition is preferably greater than 10-5, 10-3, 10-1, 0, 1, 5, 10, 25, 50, 75 or 100%. In this case, the quantity of water or liquid in the composition is preferably greater than 10-10, 10-5, 10-1, 0.1.5, 10 or 50.10 grams of water or liquid in the composition per gram of composition.

The invention also relates to the composition according to the invention, which is lyophilized or dried or dehydrated, in which the percentage by mass of cryoprotectant is preferably greater than the percentage by mass of coating in the lyophilized or dried or dehydrated composition, in which the freeze-dried or dried or dehydrated composition is preferentially more stable than the non-lyophilized or non-dried or non-dehydrated composition.

The invention also relates to the composition, where the mass percentage of the cryoprotectant is between 0.5 and 50%.

In some cases, the mass percentage of a substance included in the composition, for example the cryoprotectant, the nanoparticle core, the nanoparticle coating, the chain, the liquid or the water, is equal to or proportional to the mass of that substance divided by the mass of all substances included in the composition.

In some cases, the mass percentage of the cryoprotectant, the nanoparticle coating, the nanoparticle core, at least one chain, a liquid or water, in the composition is greater than 10-5, 10-3 , 10-1, 0, 1, 5, 10, 25, 50, 75, 90 or 99%.

In certain other cases, the mass percentage of the cryoprotectant, the nanoparticle coating, the nanoparticle core, at least one chain, a liquid or water, in the composition is less than 100 , 75, 50, 25, 10, 5, 2, 1, 0, 10-3 or 10-5%.

In one embodiment of the invention, the mass percentage of cryoprotectant in the composition according to the invention is greater than the mass percentage of nanoparticle coating, nanoparticle core, at least one chain, liquid and/or or water, in the composition.

In another embodiment of the invention, the mass percentage of cryoprotectant in the composition according to the invention is less than the mass percentage of nanoparticle coating, nanoparticle core, at least one chain, liquid and /or water, in the composition.

The invention also relates to the composition according to the invention, in which the core of the nanoparticle comprises a first center for producing or capturing free radicals C _{1 FRPC} ,
where C _1FRPC is preferentially chosen from the group consisting of:
i) a metal other than iron, such as zinc or aluminum,
And
ii) a metal oxide other than iron oxide, such as zinc oxide or aluminum oxide.

In certain cases, C _1FRPC may comprise at least one atom which is preferentially linked to at least one oxygen or iron atom of the core via at least one metallic bond.

The invention also relates to the composition according to the invention, in which the coating comprises a second center for producing or capturing free radicals C _{2 FRPC} .

The invention also relates to the composition according to the invention, in which C _2FRPC is linked to at least one atom of the coating, preferably by at least one covalent bond.

In one embodiment of the invention, the center for producing or capturing free radicals is at least one atom, a molecule, a chemical function, ionized or not, charged or not, which produces or captures at least one free radical. .

In one embodiment of the invention, a free radical is at least one atom, a molecule, an ion or a chemical function which has: i) at least one unpaired valence electron, ii) an ability to dimerize, iii) a short lifetime, and/or iv) two unpaired electrons.

In one embodiment of the invention, a free radical is at least one atom, a molecule, an ion or a chemical function which is: i) a chemically reactive species, ii) a hydroxyl radical (HO-), iii) a triplet oxygen, iv) a triplet carbene, and/or v) (꞉CH2).

In one embodiment of the invention, the radicals are generated or produced when the composition is excited in at least one of the following ways: i) by being exposed to redox reactions, ii) by being subjected to radiation , preferably ionizing or electromagnetic radiation, iii) by being heated, iv) by being exposed to electrical discharges, v) by undergoing electrolysis, vi) by being exposed to a variation in pH.

In one embodiment of the invention, the center of production of free radicals is at least one atom, a molecule, a chemical function, ionized or not, charged or not, which produces at least one free radical, preferably of such a that the quantity or concentration of free radicals present when the composition includes the free radical production center is greater than the quantity or concentration of free radicals present when the composition does not include the free radical production center, where the Comparison is preferably carried out using the same or similar excitation conditions for the composition comprising the free radical production center as for the composition not comprising the free radical production center.

In one embodiment of the invention, the free radical capture center is at least one atom, a molecule, a chemical function, ionized or not, charged or not, which captures at least one free radical, preferably of such such that the amount or concentration of free radicals present when the composition includes the free radical scavenging center is less than the amount or concentration of free radicals present when the composition does not include the free radical scavenging center, where the comparison is preferably carried out using the same or similar excitation conditions for the composition comprising the free radical capture center as for the composition not comprising the free radical capture center.

The invention also relates to the composition according to the invention, in which C _1FRPC and/or C _2FRPC is/are at least one antioxidant compound.

The invention also relates to the composition according to the invention, in which C _1FRPC and/or C _2FRPC is/are at least one oxidizing compound.

In one embodiment of the invention, the core of the nanoparticle in the composition according to the invention has at least one property chosen from the group consisting of: a) it is ferrimagnetic, b) it is composed of maghemite or magnetite or of an intermediate composition between maghemite and magnetite, c) it has a size between 0.1 and 100 nm, and d) it comprises at least one crystallographic plane.

In another embodiment of the invention, the coating of the nanoparticle in the composition according to the invention has at least one property chosen from the group consisting of: a) it has a thickness less than 10 µm, b) it comprises a number of crystallographic planes per unit area less than the number of crystallographic planes per unit area of the nucleus, c) it has a non-neutral charge, d) it is amorphous and e) it is organic, and f) it has a thickness less than 10 nm or the diameter of the nucleus.

In another embodiment of the invention, the cryoprotectant in the composition according to the invention is included in a matrix or a volume encompassing the core and the coating of the nanoparticle.

The invention also relates to the composition according to the invention, in which the nanoparticle core or core is synthesized by a living organism or cells producing nanoparticles, preferably a magnetotactic bacteria.

In one embodiment of the invention, the nanoparticles are synthesized biologically or by a living organism, designated as a living synthesizing organism, which preferably consists of or comprises at least 1, 2, 5, 10, 103, 106 or 109 cells ( s) eukaryotic(s), prokaryotic cell(s), or part of these cells. In some cases, a part of eukaryotic or prokaryotic cell(s) may be biological material originating from or produced by these cells, such as RNA, DNA, an organelle, a nucleolus, a nucleus, a ribosome, a vesicle, a rough endoplasmic reticulum, a Golgi apparatus, a cytoskeleton, a smooth endoplasmic reticulum, a mitochondrion, a vacuole, a cytosol, a lysosome, a centrosome, a cell membrane. In some cases, a biological synthesis may be defined as a synthesis involving a majority of steps, or more than 1, 2, 5 or 10 steps, or more than 1, 2, 5, 25, 50, 75 or 90% of the steps. stages, which involve chemical reactions occurring with the involvement of at least 1, 2, 10, 103, 106 or 109 living organisms, or parts of living organisms such as DNA, RNA, proteins, enzymes, lipids.

In some cases, the synthesizing living organism may be magnetotactic bacteria, types of bacteria other than magnetotactic bacteria or enzymes of certain bacteria, preferably synthesizing nanoparticles extracellularly, such as Mycobacterium paratuberculosis, Shewanella oneidensi, Geothrix fermentans, ants, mushrooms or various plants.

In yet another embodiment of the invention, the nanoparticles are synthesized or produced or crystallized or assembled or transformed into a nanoparticle by a compartment, an organelle or other biological material, such as a protein, a lipid, an enzyme, DNA or RNA, which is preferentially produced by or originates from a eukaryotic or prokaryotic cell.

In another embodiment of the invention, the nanoparticles are synthesized by or in at least one eukaryotic cell, a prokaryotic cell or a part of this cell.

In another embodiment of the invention, the nanoparticles are synthesized by or in: i) the matrix or the medium or the environment located outside of at least one eukaryotic cell, a prokaryotic cell, or a part of this cell, or ii) the extracellular matrix.

In one embodiment of the invention, the nanoparticles are synthesized by a living organism when at least 1, 2, 5, 10 or 100 step(s) of their production, such as the crystallization of iron oxide, the stabilization of the mineral iron oxide, the organization of the nanoparticles, for example into chains or aggregates, involves or is due to a living organism.

The invention also relates to nanoparticles for use, wherein the nanoparticles are magnetosomes synthesized by, derived from, extracted from, or isolated from magnetotactic bacteria.

In one embodiment of the invention, the magnetosome is synthesized by, produced by, derived from, extracted from, isolated from magnetotactic bacteria.

In one embodiment of the invention, the magnetotactic bacteria are chosen from the group consisting of: the AMB-1 strain of Magnetospirillum magneticum, the MC-1 strain of magnetotactic coccus, three strains MV-1, MV-2 and MV -4 of facultative anaerobic vibrios, Magnetospirillum magnetotacticum strain MS-1, Magnetospirillum gryphiswaldense strain MSR-1, a facultative anaerobic magnetotactic spirillum, Magnetospirillum magneticum strain MGT-1, and an obligate anaerobe, and Desulfovibrio magneticus RS- 1.

In one embodiment of the invention, a magnetotactic bacterium is defined as a bacterium capable of synthesizing magnetosomes, in which these magnetosomes are preferentially characterized by at least one of the following properties: i) they are produced intracellularly, ii) they are magnetic, iii) they include a mineral, iv) their core is preferentially composed of a metal oxide such as iron oxide, v) their core is surrounded by biological material such as lipids, proteins, endotoxins , which can preferentially be eliminated, vi) they are arranged in chains, vii) they produce heat under the application of an alternating magnetic field.

In one embodiment of the invention, the magnetosomes have one or more property(s) in common with the nanoparticles such as at least one magnetic property, size, composition, chain arrangement, charge , core, mineral, coating or crystallinity.

In one embodiment of the invention, the magnetosomes comprise the mineral part synthesized by the magnetotactic bacteria, that is to say preferably the crystallized iron oxide produced by these bacteria. In this case, the magnetosomes or the mineral parts of the magnetosomes preferably do not comprise proteins, lipids, endotoxins, or biological materials comprising carbon or do not comprise more or comprise less than 0.1, 1, 10, 30, 50 or 75% or percentage by mass of carbon, which is/are produced by these bacteria.

The invention also relates to the composition according to the invention, in which C _1FRPC and/or C _2FRPC is/are one or more photosensitizer(s), preferentially chosen from the group consisting of: 1) acridine, such such as acridine orange, acridine yellow, 2) ALA (5-aminolevulinic acid), 3) aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) aminolevulinic acid, delta acid -aminolevulinic, 5) Antihistamines, 6) Azulene, 7) Bavteriochlorine, 8) TOOKAD or TOOKAD Soluble, 9) WST-11, 10) LUZ11, 11) BC19, 12) BC21, 13) porphyrin such as Benzoporphyrin derivative monoacid ring A (BPD-MA), 14) Chlorine such as Chlorine e6, m-tetrahydroxyphenylchlorine 15) Foscan, 16) Verteporfin, 17) Monoacid A ring benzoporphyrin derivative, 18) Monoaspartyl chlorin(e6), 19) Talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrine, 23) Chlorine e6 porphrine, 24) Coal tar and derivatives, 25) Contraceptives, oral and estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview , 29) Colorants such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squaraines, 33) BODIPY dyes, 34) Phenalenones, 35) Benzophenoxazinium dyes, 36) Erythrosine, 37) Flavins, 38) Foscan , 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH), 45) Hypericin , 46) Hypocrellin, 47) ICG (Indocyanine Green), 48) Levulan, 49) MAL -methyl aminolevulinate), 50) Meta-tetra(hydroxyphenyl)chlorine (m-THPC), 51) Metvix, 52) Methylene blue, 53) Monoterpene, 54) Motexafin lutetium (Lu-Tex), 54) N-aspartyl chlorine e6 (NPe6), 55) Nanoparticle or nanomaterial, 56) Natural products or compounds, 57) Non-steroidal anti-inflammatory drugs, 58) Palladium bacteriopheophorbide (WST09), 59) Phthalocyanine-based dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phthalocyanine such as liposomal ZnPC, 65) Chloroaluminium sulfonated phthalocyanine (CASP), 66) Phthalocyanine from silicon (PC4), 67) RLP068, 68) Porfimer sodium, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-tetrakis(1-methylpyridinium-4-yl) tosylate porphyrin, 70) XF70 , 71) Protoporphyrin, 72) ALA-induced protoporphyrin IX, 73) Psoralens, 74) Quantum dots, 75) Quinones, 76) Riboflavin, 77) Rose Bengal, 78) silicon or silicon phthalocyanine (Pc4), 79) Sulfonamides, 80) Sulfonylureas, 81) Talaporfin or Talaporfin sodium, 82) Temoporfin, 82) Tetrahydropyrroles, 83) Tin ethyl etiopurpurin, 84) Titanium dioxide, 85) Toldudine blue O, 86) Transition metal compounds such as ruthenium (II), polypyridyl complexes, ruthenium, rhodium, cyclometalated bridged dimeric compounds Rh(II)-Rh(II), platinum (II), gold (III), 87) Verteporfin, 88) Compound with vulcan base such as aminovulinic acid, aminovulinic acid, 89) WST11, and 90) Xanthene.

The invention also relates to the composition according to the invention, in which C _1FRPC and/or C _2FRPC is/are one or more sonosensitizer(s), preferably chosen from the group consisting of: 1) ABS-FA, 2 ) Acrylonitrile Butadiene Styrene, 3) Styrene, 4) Folic acid, 5) AIMP NP, aminoacyl tRNA synthetase complex-interacting multifunctional protein, 6) Au Nanomaterial, 7) gold, 8) Au-MnO nanomaterial, 9) manganese oxide, 10) Anti-neoplastic drugs, 11) NSAIDs, 12) non-steroidal anti-inflammatory drugs, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) Au-doped TiO2, 17 ) carbon-based nanomaterial, 18) carbon nanotube, 19) chlorine, 20) Ce6, 21) PTX, Paclitaxel, 22) drug or chemotherapeutic compound, 23) infrared dye or IR783, 24) curcumin, 25) cyanine or Cu -Cyanine, 26) DHMS, 27) dimethylsulfide, 28) docetaxel, 29) drug or chemotherapeutic compound, 30) DOX/Mn-TPPS@RBCS, 31) doxorubicin, 32) manganese, 33) blood cell, 34) red blood cell, cell , 35) polymer, 36) elastomer, 37) erythosine or erythosine B, 38) FA or FA-OI or FA-OI NP or folic acid, 39) F3-PLGA@MB/Gd NPs, 40) poly(lactic acid- co-glycolic), 41) gadolinium, 42) Fe-TiO2 or titanium oxide, 43) Fe-VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49 ) hydrochloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysafflor yellow A, 55) Hypocrellin, Hypocrellin B, 56) IR780, 57) Levofloxacin, 58) LIP3 or Lithium phosphide , 59) Lithium, 60) Liposome or liposomal nanomaterial, 61) Lomefoxacin,, 62) MG@P NPs, 63) MnP or Manganese peroxidase, 64) MnTTP-HSAs, 65) metal-porphyrin complex wrapped in HSA, 66) albumin , 67) MnWOx, 68) MnWOx-PEG, 69), PEG, 70) bimetallic oxide, 71) Mn (III)-HFs, 72) managen, hemoporphin, 73) Nano-beads, 74) Noble metal nanomaterial, 75 ) OI NP or indyocyanine oxygen, 76) Phthalocyanines, 77) PIO or Pioglitazone, 78) Polymeric nanomaterial, 79) Porphyrin, 80) Pt-doped TiO2, 81) R837, 82) Rose Bengal, 83) Sparfloxacin,, 84) TAPP or 5,10,15,20-tetrakis(4-aminophenyl)porphyrin, 85) TiO2 or titanium dioxide nanomaterial, 86) TCPP, tris(1-chloro-2-propyl) isomer or phosphate 87) TPI or polyimide thermoplastic or thermoplastic polymer, 88) TPZ or Tirapazamine, 89) Transition metal oxide, 90) Janus nanoparticle or nanoparticle, and 91) Xanthones.

The invention also relates to the composition according to the invention, in which C _1FRPC and/or C _2FRPC is/are radiosensitizers, preferably chosen from the group consisting of: 1) AMG102, 2) AQ4N, 3) Apaziquone (E09 ), 4) Bromodeoxyuridine, 5) Carbogen, 6) Cetuximab, 7) Chemotherapeutic drug or compound, 8) Chlorpromazine, 9) C-reactive peptide, 10) Curcumin, 11) Diamide, 12) Diethylmaeate,, 13) Dihydroartemisinin, 14 ) Docetaxel, 15) ECI301, 16) Etanidazole, 17) Fludarabine, 18) 5-Fluorouracil, 19) Fluorodeoxyuridine, 20) Gadolynium, 21) Gemcitabine, 22) HER-3 ADC, 23) HSP, 24) Hydrogen peroxide , 25) Hydroxyurea, 26) Hyperbaric oxygen, 27) Hyperthermia, 28) Hypoxic cellular cytotoxic agent, 29) Irinotecan, 30) Radiosensitive lanthanide-based metal-phenolic network, 31) Lidocaine, 32) Lododeoxyuridine, 33) Metronidazole, 34) misonidazole, 35) etanidazole, 36) nimorazole, 37) N-Ethylmalemide, 38) malmeide, 39) ethylmalmeide, 40) Nanomaterials such as those consisting or composed at least partially or totally of gold, silver, bismuth , gadolinium, polysiloxane matrix and chelates of gadolinium, hafnium, tantalum, zinc, gadolinium, germanium, chromium, praseodymium, silicon, iron, platinum, cobalt, manganese, magnesium, iron, titanium, carbon nanotube, quantum dot, nanorad, triflate or metal oxide, 41) Nelfinavir, 42) Nicotinamide, 43) Nimotuzumab, 44) RNA, or miRNA, or miR -201, or miR-205, or miR-144-5p, or miR-146a-5p, or miR-150, or miR-99a, or miR-139-5p, or miR-320a, 45) Active agent on membrane , 46) Mitomycin-C or Mitomycin, 47) Motexafin, 48) NBTXR3, 49) Oligonucleotide, 50) Paclitaxel, 51) Papaverine or papaverine hydrochloride, 52) Paraxonase-2, 53) Pocaine, 54) Porfiromycin (POR), 55) Protein, 56) Peptide, 57) Nucleosides or radiosensitizing compounds, 58) Resveratrol, 59) RRx-001, 60) SiRNa, 61) Sulfhydric group suppressants, 62) SYM004, 63) Texaphyrins, 64) TH-302, and 65) Tirapazamine.

In one embodiment of the invention, C _1FRPC and C _2FRPC act in synergy, that is to say that the photo-sensitization or sono-sensitization or radio-sensitization force of the nanoparticle comprising C _1FRPC and C _2FRPC is greater than the photo-sensitization or sono-sensitization or radio-sensitization force of the nanoparticle comprising only C _1FRPC or C _{2 FRPC} .

In certain cases, the strength of photosensitization or sono-sensitization or radiosensitization is the concentration or quantity of radical species, preferably free radical species, produced or captured by the at least one photosensitizer, radio- sensitizer, sono-sensitizer, center of capture or production of free radicals, or composition, or is or is proportional to the strength or intensity of the acoustic wave, ultrasound, radiation, light, electromagnetic radiation, preferably applied to the photosensitizer, the radio-sensitizer, the sono-sensitizer, the free radical capture or production center or the composition.

In one embodiment of the invention, C _1FRPC and C _2FRPC act in an antisynergistic manner, that is to say that the strength of photosensitization or sono-sensitization or radio-sensitization of the nanoparticle comprising C _1FRPC and C _2FRPC is less than the photo-sensitization or sono-sensitization or radio-sensitization force of the nanoparticle comprising only C _1FRPC or C _{2 FRPC} .

In one embodiment of the invention, the composition is introduced or administered to or in the part of the body, preferably of an animal or a human. Preferably, after its introduction or its administration, the nanoparticle first comprises C _1FRPC and C _2FRPC for a time t1 and then comprises C _1FRPC for a time t2, where t2 follows t1.

In some cases, t1 and/or t2 may be longer than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10 or 103 second(s) or minute(s).

In some other cases, t1 and/or t2 may be shorter than 1010, 105, 103, 10, 0, 1, 5, 10-1 or 10-3 second(s) or minute(s).

In some cases, t2 may be separated from t1 by more than 10-10, 10-5, 10-3, 10-1, 0, 1, 5, 10 or 103 second(s) or minute(s).

In some other cases, t2 may be separated from t1 by less than 1010, 105, 103, 10, 0, 1, 5, 10-1 or 10-3 second(s) or minute(s).

The invention also relates to the composition according to the invention, where the composition is or comprises: i) a medical device, ii) a medicine, iii) a pharmaceutical product or preparation, iv) a medical product or preparation, iv) a biological product or preparation, vi) an adjuvant, vii) an excipient, viii) an active ingredient, ix) a vaccine or vaccine component, vi) a product, vii) a suspension, viii) a suspension or composition or a freeze-dried preparation.

In one embodiment, the composition according to the invention is used for/in: i) the treatment of an infection, ii) the treatment of a viral disease, iii) the treatment of a disease, preferably a cancerous disease , iv) radiotherapy, v) photodynamic therapy, and/or vi) sonodynamic therapy.

In some cases, the disease may be due to the poor functioning of an organ or part of the body preferentially in an individual.

In some cases, the treatment may be therapy and/or diagnosis of a disease or cosmetic treatment.

In certain cases, the treatment may induce the death, destruction, denaturation or inactivation of at least one biological material, such as a cell, preferably a pathological cell, an RNA, a DNA, a protein, a lipid or an enzyme, cell death may occur by apoptosis or necrosis.

The invention also relates to nanoparticles to be used according to the invention, the disease is chosen from the group consisting of: a disease associated with a proliferation of cells different from cell proliferation in a healthy individual, a disease associated with the presence of cells pathological conditions in the body part, a disease associated with the presence of a pathological site in an individual or body part, a disease or disorder or dysfunction of the body part, a disease associated with the presence of radio cells -resistant or acoustic, an infectious disease, an autoimmune disease, a neuropathology, a cancer, a tumor, a disease comprising or due to at least one cancerous or tumor cell, a skin condition, an endocrine disease, a disease or a eye disorder, intestinal disease, communication disorder, genetic disorder, neurological disorder, voice disorder, vulvovaginal disorder, liver disorder, heart disorder, heating disorder, speech disorder mood, anemia, preferably iron anemia, and personality disorder.

In some cases, the disease or disorder may be the disease or disorder of the individual or part of or belonging to the body, or the disease or disorder from which the individual suffers.

In one embodiment of the invention, the cancer or tumor is chosen from the group consisting of: cancer of an organ, cancer of the blood, cancer of a system of a living organism, cancer of adrenal, anal cancer, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colon cancer /rectum, endometrial cancer, esophageal cancer, eye cancer, gallbladder cancer, heart cancer, kidney cancer, larynx and hypopharynx cancer, leukemia , liver cancer, lung cancer, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, oral cavity and oropharynx cancer, osteosarcoma cancer, breast cancer ovarian, pancreatic cancer, penile cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, stomach cancer, testicular cancer, breast cancer thymus, thyroid cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, waldenstrom macroglobulinemia wilms tumor, castleman disease ewing family tumor, gastrointestinal carcinoid tumor, stromal tumor gastrointestinal syndrome, pituitary tumor myelodysplastic syndrome, and cancerous disease such as gestational trophoblastic disease, Hodgkin's disease, Kaposi's sarcoma, malignant mesothelioma, and multiple myeloma.

The invention also relates to a method for the treatment of anemia or anemic disease or to nanoparticles, in particular magnetosomes, for use in the treatment of anemic disease, preferably iron anemic disease, in which the Magnetosomes are administered to an individual's body part, preferably to reduce or stop anemia.

The invention also relates to a method for the treatment of an anemic disease, in which this disease is selected from the group consisting of: iron or metal deficiency anemia, vitamin deficiency anemia, anemia chronic disease, aplastic anemia, anemia associated with bone marrow disease, hemolytic anemia, sickle cell anemia, thalassemia, pernicious anemia, Fanconi anemia, sideroblastic anemia, congenital dyserythropoietic anemia (CDA), Diamond Blackfan anemia and megaloblastic anemia.

In some cases, anemia is a decrease in the total amount of red blood cells (RBCs) or hemoglobin in the blood, or a decrease in the blood's ability to carry oxygen.

The invention also relates to a process for manufacturing the composition according to the invention, which comprises at least one of the following steps:
- Step 1 of amplification of the magnetotactic bacteria in at least one medium, preferably chosen from a pre-growth, growth and/or feed batch medium (a), preferably comprising:
1) the compounds necessary for the growth of magnetotactic bacteria and/or the production of magnetosomes, which are preferentially chosen from the group consisting of:
-a carbon source, which is preferably chosen from the group consisting of: at least one compound comprising at least one carbon atom, lactic acid, Na lactate, acetate, glycolate, glucose, pyruvate , succinate, carbon dioxide, glycerol and their combinations, at a concentration preferably between 1 nM and 2 Mol/L;
a source of iron preferably chosen from the group consisting of: at least one compound comprising at least one iron atom, iron citrate, iron quinate, iron chloride, iron sulfate, FeCl3, and their combinations , at a concentration preferably between 1 nM and 2.10-3 Mol/L;
a source of nitrogen preferably chosen from the group consisting of: at least one compound comprising at least one nitrogen atom, a nitrate salt, gaseous nitrogen, ammonium, ammonia, a salt of ammonium, urea, an amino acid, gaseous ammonia and combinations thereof, at a concentration preferably between 1 nM and 4 Mol/L;
-a source of oxygen preferably chosen from the group consisting of: at least one compound comprising at least one oxygen atom, oxygen or air or compressed air, preferably in gas form, the source of oxygen being in certain cases bubbled or introduced into the growth medium, at a gas flow rate which is preferably between 5 mL of gas per minute and 50,000 mL of gas per minute;
a source of phosphate consisting preferably of at least one compound comprising at least one phosphate atom, at a concentration preferably between 1 nM and 2.10-1 Mol/L;
a source of potassium consisting preferably of at least one compound comprising at least one potassium atom, at a concentration preferably between 1 nM and 2.10-1 Mol/L;
a source of sulfur or sulfate preferably consisting of at least one compound comprising at least one sulfur or sulfate atom, at a concentration preferably between 1 nM and 4.10-1 Mol/L;
a source of manganese preferentially consisting of at least one compound comprising at least one manganese atom, at a concentration preferably between 1 nM and 4.10-1 Mol/L;
a source of vitamin preferably chosen from the group consisting of: at least one compound comprising at least one vitamin, biotin, calcium, pantothenate, folic acid, inositol, nicotinic acid, p acid -aminobenzoic acid, pyridoxine HCl, riboflavin, thiamine, thiamine HCL and their derivatives and their combinations, at a concentration preferably between 1 nM and 10-4 Mol/L, and
a source of calcium consisting preferably of at least one compound comprising at least one calcium atom, at a concentration preferably between 1 nM and 10-1 Mol/L.
2) at least one compound necessary for doping the magnetosomes with C _1FRPC or a metal other than iron, preferably zinc or aluminum, for example a source of zinc, preferably zinc sulfate or zinc citrate or zinc chlorate or zinc quinate.
- Step 2 of extracting magnetosomes from magnetotactic bacteria and/or purifying the extracted magnetosomes, preferably by heating, to obtain magnetosome minerals preferably comprising a mass percentage of organic matter originating from magnetotactic bacteria of less than 100 , 50 or 1%,
- Step 3 of coating the magnetosome minerals with a coating material preferably comprising the compound C _2FRPC by mixing the magnetosome minerals with the coating material, where the mixing is preferably carried out under at least one of the following conditions:
i) under sonication,
ii) under the application of radiation,
under temperature variation,
under pH changes,
under adjustment of the redox potential,
preferably using a ratio between the quantity or mass of the minerals of the magnetosome and the quantity or mass of the coating material, preferably of compound D, which is preferably adjusted or varied or greater than 1,
- step 4 consisting of adding at least one cryoprotectant to the coated magnetosome minerals obtained at the end of step 3,
- Step 5 of freeze-drying or dehydration or drying or desiccation of the composition obtained at the end of step 4,
- Step 6 of resuspension of the lyophilized composition obtained in step 5, preferably in water or in another liquid.

The invention also relates to the process according to the invention, in which the concentration of the zinc source, preferably zinc citrate or zinc sulfate, is between 1 and 100 µM, preferably 2 and 50 µM, more preferably 5 and 20 µM.

The invention also relates to the method according to the invention, in which the nanoparticles manufactured according to the method comprise a quantity of metal other than iron, preferably zinc or aluminum, which is incorporated in or included in the metal core is: i) between 0.1 and 100 mg of metal other than iron included in the core per gram of iron included in the core, ii) preferably between 0.5 and 10 mg of metal other than iron included in the core per gram of iron included in the core, iii) more preferably between 1 and 5 mg of metal other than iron included in the core per gram of iron included in the core.

In one embodiment of the present invention, the nanoparticle is or belongs to or is included in the group of nanoparticles chosen from: a nanosphere, a nanocapsule, a dendrimer, a carbon nanotube, a lipid/solid nanoparticle, a nanoparticle with based on lipid or protein or DNA or RNA, a nanoparticle with an internal aqueous environment surrounded by a layer, preferably a stabilizing layer, more preferably a phospholipid layer, a multilayer nanoparticle, a polymer nanoparticle, a quantum dot, a metal nanoparticle, a micelle or polymer nanoparticle, a carbon-based nanostructure, a nanobubble, a nanosome, a pharmacyte, a niosome, a nanopore, a microbivore, a liposome, a virus, preferably recombinant, a nanoparticle plant, an antibody and a vesicle.

In another embodiment of the present invention, the nanoparticle is not or does not belong to or is not included in at least one nanoparticle belonging to the group of: a nanosphere, a nanocapsule, a dendrimer, a nanotube of carbon, a lipid/solid nanoparticle, a nanoparticle based on lipid or protein or DNA or RNA, a nanoparticle with an internal aqueous environment surrounded by a layer, preferably a stabilizing layer, more preferably a layer phospholipid, a multilayer nanoparticle, a polymer nanoparticle, a quantum dot, a metal nanoparticle, a micelle or polymer nanoparticle, a carbon-based nanostructure, a nanobubble, a nanosome, a pharmacyte, a niosome, a nanopore, a microbivore, a liposome, a virus, preferably recombinant, a plant nanoparticle, an antibody and a vesicle.

In some cases, the nanoparticle may be in a liquid, gaseous or solid state, preferably before, during or after its presence or administration to the body part.

In other cases, the nanoparticle may not be in one or both of the liquid, gas, or solid states, preferably before, during, or after its presence or administration to the body part.

In still other cases, the nanoparticles may be assimilated to or included in a ferrofluid, a chemical or biological ferrofluid, where the chemical and biological ferrofluids are iron-containing fluids, preferably forming nanoparticles, which are manufactured by a synthesis chemical or biological, respectively.

In still other cases, the ferrofluid or the assembly of nanoparticles may comprise the nanoparticles and an excipient, a solvent, a matrix, a gel, which preferentially allows the administration of the nanoparticles to the individual or to the part of the body. .

In still other cases, the nanoparticle may comprise a synthetic material and/or a biological material and/or an inorganic material and/or an organic material.

In one embodiment of the invention, the nanoparticle(s) are or designate: i) a suspension of nanoparticles, ii) a composition comprising nanoparticles, iii) an assembly of nanoparticles, iv) a region of nanoparticles, v) the mineral part or the core of the nanoparticle, vi) the organic part of the nanoparticle, vii) the inorganic part of the nanoparticle, viii) or the coating of the nanoparticle.

In one embodiment of the invention, the nanoparticle(s) or nanoparticle(s) represent or are an assembly or suspension or composition of more than or comprising more than 10-100, 10-50, 10-10, 10 -5, 10-1, 1, 10, 102, 103, 105, 1010, 1020 or 1050 nanoparticle(s) or mg of nanoparticle(s) or mg of iron included in the nanoparticle(s) or mg of nanoparticle(s) ) per cm3 or mg of nanoparticle(s) per cm3 of the body part or mg of iron included in the nanoparticle(s) per cm3 or mg of iron included in the nanoparticle(s) per cm3 of the body part. In some cases, an assembly or suspension or composition comprising a large number of nanoparticles can be used to induce or produce an increase in temperature, a radical or reactive species, or the dissociation of a compound from the nanoparticles.

In another embodiment of the invention, the nanoparticle(s) or the nanoparticle(s) represent or are an assembly or a suspension or a composition of less than or comprising less than 10100, 1050, 1020, 1010, 105, 102, 10 , 1, 5, 2, 1, 10-1, 10-5, 10-10 or 10-50 nanoparticle(s) or mg of nanoparticle(s) or mg of iron included in the nanoparticle(s) or mg of nanoparticle (s) per cm3 or mg of nanoparticle(s) per cm3 of the body part or mg of iron included in the nanoparticle(s) per cm3 or mg of iron included in the nanoparticle(s) per cm3 of the body part . In some cases, an assembly or suspension or composition of nanoparticles comprising a low number of nanoparticle(s) can be used to prevent toxicity.

In one embodiment of the invention, the nanoparticle(s) or set of nanoparticles may represent or be the region, also referred to as nanoparticle region, volume, surface area, length, which comprises the nanoparticles or where the nanoparticles are located. . In some cases, the volume of the region occupied by the nanoparticles in the body part is designated as the nanoparticle region.

In some cases, the nanoparticle region may be the volume occupied by an array of nanoparticles in the body part, where the nanoparticles are preferentially separated by less than 109, 106, 103, or 10 nm.

In some cases, nanoparticle assembly is a more general term than nanoparticle region, which could refer to any type of nanoparticle assembly, before, during or after delivery of nanoparticles to or into the body part.

In some cases, the separation distance between nanoparticles within the nanoparticle assembly or nanoparticle region may correspond to the average or maximum distance separating the nanoparticles within that assembly.

In certain cases, the distribution of separation distances between nanoparticles can highlight the presence of a minority of nanoparticles, that is to say preferentially less than 50, 10, 1, 10-2 or 10-5% of the total number of nanoparticles in the individual, with either small separation distances, that is to say separation distances preferentially less than 10, 10, 10 or 10 nm, or large separation distances, it is that is to say separation distances preferably greater than 10, 10 or 10 nm. that is to say separation distances preferably less than 109, 106, 103 or 10 nm, i.e. with large separation distances, c that is to say separation distances preferably greater than 109, 106, 103 or 10 nm. In this case, the presence of this minority of nanoparticles is preferably not taken into account to estimate the average or maximum separation distance between the nanoparticles.

The invention also relates to nanoparticles for use according to the invention, in which the nanoparticles are crystallized, metallic or magnetic.

In one embodiment of the invention, the nanoparticles are crystallized. In this case, they preferably have more than or at least 1, 2, 10, 102, 103, 106 or 109 crystallographic plane(s) or regular atomic arrangement(s), observable preferentially by electron microscopy.

In one embodiment of the invention, the nanoparticles are metallic. In this case, they contain at least 1, 10, 103, 105 or 109 metal atom(s) or contain at least 1, 10, 50, 75 or 90% of metal atoms, this percentage possibly being the ratio between the number or mass of metal atoms in the nanoparticle divided by the total number or mass of all atoms in the nanoparticle. The nanoparticles, preferably metal oxide nanoparticles, can also contain at least 1, 10, 103, 105 or 109 atom(s) of oxygen, or contain at least 1, 10, 50, 75 or 90% of oxygen atoms, where this percentage can be the ratio of the number or mass of oxygen atoms in the nanoparticles divided by the total number or mass of all atoms in the nanoparticles.

In another embodiment of the invention, the metal or the metal atom is chosen from the list consisting of: Lithium, Beryllium, Sodium, Magnesium, Aluminum, Potassium, Calcium, Scandium, Titanium, Vanadium, Chromium, Manganese, Iron, Cobalt, Nickel, Copper, Zinc, Gallium, Rubidium, Strontium, Yttrium, Zirconium, Niobium, Molybdenum, Technetium, Ruthenium, Rhodium, Palladium, Silver, Cadmium, Indium, Tin, Cesium, Barium, Lanthanum, Cerium, Praseodymium, Neodymium, Promethium, Samarium, Europium, Gadolinium, Terbium, Dysprosium, Holmium, Erbium, Thulium, Ytterbium, Lutetium, Hafnium, Tantalum, Tungsten, Rhenium, Osmium, Iridium, Platinum, Gold, Mercury, Thallium, Lead, Bismuth, Polonium, Francium, Radium, Actinium, Thorium, Protactinium, Uranium, Neptunium, Plutonium, Americium, Curium, Berkelium, Californium, Einsteinium, Fermium, Mendelevium, Nobelium, Lawrencium, Rutherfordium, Dubnium, Seaborgium, Bohrium, Hassium, Meitnerium, Darmstadtium, Roentgenium, Copernicium, Nihonium, Flerovium, Moscovium, and Livermorium or Livermorium atom.

In another embodiment of the invention, the nanoparticle contains less than 1, 10, 103, 105 or 109 metal atom(s) or contains less than 1, 10, 50, 75 or 90% atoms metals, where this percentage can be the ratio of the number or mass of metal atoms in the nanoparticle divided by the total number or mass of all atoms in the nanoparticle. It may also contain less than 1, 10, 103, 105 or 109 atom(s) of oxygen, or contain less than 1, 10, 50, 75 or 90% of oxygen atoms, this percentage may be the ratio between the number or mass of oxygen atoms in the nanoparticle divided by the total number or mass of all atoms in the nanoparticle.

In one embodiment of the invention, the nanoparticle is magnetic when it has a magnetic behavior or property, where the magnetic behavior or property is preferentially chosen from the group consisting of a diamagnetic, superparamagnetic, paramagnetic behavior or property. , ferromagnetic and ferrimagnetic.

In some cases, the magnetic behavior or property may be observed or exist at a temperature that is lower than: i) 105, 103, 500, 350, 200, 100, 50, 20, 10, 1, 0.5 or 1 K (Kelvin), ii) the Curie temperature, or iii) the blocking temperature.

In other cases, the magnetic behavior or property may be observed or exist at a temperature above: i) 0.5, 1, 10, 20, 50, 100, 200, 350, 500, 103 or 105 K, ii) the Curie temperature, or iii) the blocking temperature.

In still other cases, the magnetic behavior or property may be observed or exist at a temperature between 10-20 and 1020 K, or between 0.1 and 1000 K.

In one embodiment of the invention, the nanoparticles present or are characterized by at least one of the following properties: i) the presence of a core, preferably magnetic, preferably mineral, preferably composed of a metal oxide such as iron oxide, more preferably maghemite or magnetite, or an intermediate composition between maghemite and magnetite, ii) the presence of a coating which surrounds the core and preferentially prevents the aggregation of the nanoparticles, preferentially allowing the administration of the nanoparticles in an organism or in the part of the body or stabilizing the core of the nanoparticles, the thickness of the coating can preferably be between 0.1 nm and 10 μm, between 0.1 nm and 1 μm, between 0.1 nm and 100 nm , between 0.1 nm and 10 nm, or between 1 nm and 5 nm, iii) magnetic properties leading to diamagnetic, paramagnetic, superparamagnetic, ferromagnetic or ferrimagnetic behavior, iv) a coercivity greater than 0.01, 0.1, 1, 10 , 100, 103, 104, 105, 109 or 1020 Oe, v) a ratio between the remanent magnetization and the saturation magnetization greater than 0.01, 0.1, 0.2, 0.3, 0.4 , 0.5, 0.75, 0.9 or 0.99, vi) a saturation magnetization greater than 0.1, 1, 5, 10 or 50 emu/g, vii) magnetic properties such as coercivity, remanent magnetization and saturating magnetization, preferably measured or observed at a temperature greater than 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K, viii ) crystallinity, that is to say nanoparticles, preferably measured or observed at a temperature greater than 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K. that is to say nanoparticles preferably having at least 1, 2, 5, 10 or 100 crystal plane(s), observable(s) or measured(s) preferably by microscopy electronic, ix) the presence of a unique domain, 120, 150 or 200 nm, xi) a size between 0.1 nm and 10 μm, between 0.1 nm and 1 μm, between 0.1 nm and 100 nm, between 1 nm and 100 nm, or between 5 nm and 80 nm, xii) non-pyrogenicity or apyrogenicity, which preferably means that the nanoparticles have an endotoxin concentration less than 1020, 10000, 1000, 100, 50, 10, 5, 2 or 1 EU (endotoxin unit) per mg of nanoparticle or per mg of iron included in the nanoparticles, or which means that the nanoparticles do not trigger a fever or an increase in whole body temperature greater than 100, 50, 6.6, 5, 3, 2 or 1 C after their administration to a living organism or part of the body, xiii) synthesis by a living synthesizing organism, preferably by bacteria, xiv) chemical synthesis, xv) the presence of less than 50, 25, 15, 10, 5, 2 or 1% of organic or carbonaceous matter coming from the living synthesizer organism, xv), the presence of more than 99, 95, 80, 70, 60, 50 or 25% of mineral matter coming from the synthetic living organism, or xvi) a specific absorption rate (SAR) which is greater than 1, 10, 1000 or 104 Watt per gram of nanoparticle, preferably measured under the application of an alternating magnetic field of intensity preferably greater than 0.1, 1, 10 or 100 mT, and/or frequency greater than 1, 10, 100 or 1000 KHz, alternatively measured preferentially under the application of the acoustic wave, alternatively under the application of a radiation such as acoustic or light electromagnetic radiation.

In another embodiment of the invention, the nanoparticles have or are characterized by at least one of the following properties: i) a coercivity less than 0.01, 0.1, 1, 10, 100, 103, 104, 105 , 109 or 1020 Oe, ii) a ratio between the remanent magnetization and the saturating magnetization less than 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.75, 0, 9 or 0.99, iii) a saturation magnetization less than 0.1, 1, 5, 10, 50, 200, 1000 or 5000 emu/g, iv) magnetic properties measured or observed preferably at a temperature lower than 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K, v) a size less than 0.1, 0.5, 1.5, 10, 15, 20, 25, 30, 50, 60, 70, 80, 100, 120, 150 or 200 nm, vi) the presence of more than 50, 25, 15, 10, 5, 2 or 1% organic matter or carbon dioxide coming from the living synthesizing organism, vii) the presence of less than 99, 95, 80, 70, 60, 50 or 25% of mineral matter coming from the living synthesizing organism, or xi) an absorption rate specific (SAR) less than 1, 10, 1000 or 104 Watt per gram of nanoparticle, preferably measured under the application of an alternating magnetic field of intensity preferably less than 0.1, 1, 10, or 100, 200, 500 , 103 or 105 mT, and/or of frequency preferably less than 1, 10, 100, 103, 105 or 109 KHz, alternatively measured preferentially under the application of the acoustic wave, alternatively under the application of radiation such as acoustic or luminous electromagnetic radiation.

In some cases, the mineral may be the part of the nanoparticle or magnetosome that does not include organic matter or that includes a low mass percentage of organic matter, preferably less than 100, 99, 50, 20, 10, 5 , 1, 10-1 or 10-2 percent or mass percentage of organic matter. The mineral is preferably the core of the nanoparticle.

In certain other cases, the mineral may include a mass percentage of organic matter greater than 0, 10-50, 10-10, 10-2, 10-1 or 1 percent or mass percentage of organic matter. This may be the case when the purification step fails to remove the organic material or when the organic material is added to the mineral after the purification step.

In some cases, the nanoparticles can be surrounded by a coating. The coating may consist of a synthetic, organic or inorganic material or of a substance comprising a function chosen from the group consisting of carboxylic acids, phosphoric acids, sulfonic acids, esters, amides, ketones, alcohols, phenols, thiols, amines, ethers, sulfides, acid anhydrides, acyl halides, amidines, amides, nitriles, hydroperoxides, imines, aldehydes and peroxides. In some cases, the coating may consist of carboxy-methyl-dextran, citric acid, phosphatidylcholine (DOPC), or oleic acid. In certain cases, the coating can allow the dispersion of the nanoparticles in a matrix or a solvent such as water, preferably without aggregation or sedimentation of the nanoparticles. In some cases, the coating may allow internalization of nanoparticles into cells. In other cases, the coating can allow: i) to bind two or more nanoparticles together, preferably in a chain, ii) to prevent aggregation of the nanoparticles and/or, iii) to obtain a uniform distribution nanoparticles.

In one embodiment of the invention, the nanoparticles are non-pyrogenic. The non-pyrogenic nanoparticles are preferably: i) comprise less than 10100, 1050, 1020, 108, 105, 103, or 10 EU (endotoxin unit) or EU per cm3 of the body part or EU per mg of nanoparticle or EU per cm3 of body part per mg of nanoparticle, or ii) induce an increase in temperature of the individual or body part lower than 105, 103, 102, 50, 10, 5, 4, 3, 2 or 1°C, preferably above the physiological temperature, preferably before, after or without the application of the acoustic wave or radiation to the nanoparticle.

In one embodiment of the present invention, the nanoparticle or the compound is composed of or comprises a chemical element from the families chosen from the group consisting of: metals (alkali metal, alkaline earth metal, transition metals), semi-metal , non-metal (halogen element, noble gas), chalcogen elements, lanthanide and actinide.

In another embodiment of the invention, the nanoparticle or the compound is composed of or comprises a chemical element chosen from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, cesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, lanthanides, actinides, titanium, zirconium, hafnium, rutherfordium, vanadium, niobium, tantalum, dubnium, chromium, molybdenum, tungsten, seaborgium, manganese, technetium, rhenium, bohrium, iron, ruthenium, osmium, hessium, cobalt, rhodium, iridium, meitherium, nickel, palladium, platinum, darmstadtium, copper, silver, gold, roentgenium, zinc, cadmium, mercury, copernicum, boron, aluminum, gallium, indium, thallium, ununtrium, carbon, silicon, germanium, tin, lead, fleovium, nitrogen, phosphorus, arsenic, antimony, bismuth, ununpentium, oxygen, sulfur, selenium, tellurium, polonium, livermorium, fluorine, chlorine, bromine, iodine, astatine, ununseptium, helium, neon, argon, krypton, xenon, radon, ununoctium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium, actinium, thorium, proctactinium, uranium, neptunium, plutonium, americium, curium, berkelium, californium, einsteinium, fermium, mendelevium, nobelium and lawrencium.

In some cases, the nanoparticle or compound may also be composed of or include an alloy, a mixture, or an oxide of this chemical element(s).

In some cases, the nanoparticle or compound may be composed of more than 10-50, 10-20, 10-10, 10-5, 10-2, 1, 5, 10, 50, 75, 80, 90, 95 or 99% of one or more of these element(s), where this percentage may represent the mass or number of this chemical element(s) included in the nanoparticle or compound divided by the total number or total mass of all chemical elements included in the nanoparticle or compound or by the total mass of the nanoparticle or compound.

In certain other cases, the nanoparticle or compound may be composed of or include less than 10-50, 10-20, 10-10, 10-5, 10-2, 1, 5, 10, 50, 75, 80, 90, 95 or 99% of one or more of these chemical element(s).

In still other cases, this or these chemical elements are included inside the nanoparticle or compound, or on the surface of the nanoparticle or compound, or in the mineral or core of the nanoparticle or compound, or in the coating of the nanoparticle or compound.

In one embodiment of the present invention, the nanoparticle or the compound is not composed of or does not comprise at least one chemical element belonging to the family chosen from the group consisting of: metals (alkaline metal, alkaline earth metal , transition metals), semi-metals, non-metals (halogen element, noble gas), chalcogenic elements, lanthanides, actinides.

In another embodiment of the invention, the nanoparticle or the compound is devoid of or does not comprise at least one chemical element chosen from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, cesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, lanthanides, actinides, titanium, zirconium, hafnium, rutherfordium, vanadium, niobium, tantalum, dubnium, chromium, molybdenum, tungsten, seaborgium, manganese, technetium, rhenium, bohrium, iron, ruthenium, osmium, hessium, cobalt, rhodium, iridium, meitherium, nickel, palladium , platinum, darmstadtium, copper, silver, gold, roentgenium, zinc, cadmium, mercury, copernicum, boron, aluminum, gallium, indium, thallium, ununtrium, carbon, silicon, germanium, tin, lead, fleovium, nitrogen, phosphorus, arsenic , antimony, bismuth, ununpentium, oxygen, sulfur, selenium, tellurium, polonium, livermorium, fluorine, chlorine, bromine, iodine, astatine, ununseptium, helium, neon, argon, krypton, xenon, radon, ununoctium, lanthanum, cerium, praseodymium , neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium, actinium, thorium, proctactinium, uranium, neptunium, plutonium, americium, curium, berkelium, californium, einsteinium, fermium, mendelevium , nobelium and lawrencium.

In another embodiment of the invention, the nanoparticle or compound is not composed of or does not include an alloy, mixture or oxide of this chemical element(s).

In one embodiment of the invention, the nanoparticle is defined as a particle whose size in one dimension is greater than 10-1, 1, 2, 5, 10, 20, 50, 70, 100, 200 or 500 nm . A large nanoparticle may have greater coercivity and/or remanent magnetization and/or may absorb the energy or power of the acoustic wave more strongly or more efficiently than a small nanoparticle. In some cases, the amount of energy or power absorbed by a nanoparticle is increased by a factor greater than 1.001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 103, 105 or 107 by increasing the size of the nanoparticle by a factor greater than 1.001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 103, 105 or 107.

In another embodiment of the invention, the nanoparticle is defined as a particle whose size in one dimension is less than 104, 103, 102, 10.1 or 10-1 nm. A small nanoparticle can be more easily administered, for example intravenously, or can help avoid certain toxic effects, such as embolism.

In yet another embodiment of the invention, the size of the nanoparticles is between 10-2 and 1020 nm, 10-2 and 104 nm, between 10-1 and 103 nm, or between 1 and 102 nm. This may be the case when the nanoparticle or the assembly of nanoparticles has a well-defined, preferably narrow, size distribution.

In yet another embodiment of the invention, the size distribution of the nanoparticles is less than 1000, 100, 75, 50, 25, 10, 5, 2 or 1 nm. A narrow nanoparticle size distribution may be desired to prevent aggregation, or to promote chain organization of the nanoparticles.

In yet another embodiment of the invention, the size distribution of the nanoparticles is greater than 1000, 100, 75, 50, 25, 10, 5, 2 or 1 nm. A wide nanoparticle size distribution can in some cases allow nanoparticles to be removed more quickly.

In another embodiment of the invention, the nanoparticle has a surface charge greater than -200, -100, -50, -10, -5, 0.1, 1, 2, 5, 10, 50 or 100 mV, preferably at a pH less than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14. Preferably, a nanoparticle can have a significant surface charge at low pH when surrounded by a coating allowing such a charge to be achieved without being destroyed.

In another embodiment of the invention, the nanoparticle has a surface charge that is less than -200, -100, -50, -10, -5, 0.1, 1, 2, 5, 10, 50 or 100 mV, preferably at a pH greater than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14. A nanoparticle may have a low charge surface at high pH when surrounded by a coating which allows such a charge to be achieved without being destroyed.

In another embodiment of the invention, the nanoparticle has a surface charge of between +200 and -200 mV, +100 and -100 mV, +50 and -50 mV, +40 and -40 mV, +20 and -20, +10 and -10 mV, or between +5 and -5 mV, preferably at a pH lower than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12, 13 or 14.

In another embodiment of the invention, the nanoparticle has a surface charge of between +200 and -200 mV, +100 and -100 mV, +50 and -50 mV, +40 and -40 mV, +20 and -20, +10 and -10 mV, or between +5 and -5 mV, preferably at a pH greater than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12, 13 or 14.

In another embodiment of the invention, the nanoparticle has a weight or mass, preferably expressed in units such as gram (g), kilogram (kg), or milligram (mg). One gram of nanoparticle may be one gram of metal such as iron included in the nanoparticle. The mass or weight of the nanoparticle can correspond to the mass or weight of a nanoparticle or to the mass or weight of an assembly of nanoparticles.

In one embodiment, the mass of the nanoparticle is greater than 10-20, 10-10, 10-5, 10-2, 1, 10, 103, 109 or 1020 grams. In some cases, a large mass of nanoparticles may be desired to increase the amount of acoustic wave energy absorbed by the nanoparticle.

In one embodiment, the mass of the nanoparticle is less than 10-20, 10-10, 10-5, 10-2, 1, 10, 103, 109 or 1020 grams. In some cases, a low nanoparticle mass may be desired to prevent or minimize nanoparticle toxicity.

In one embodiment of the invention, the nanoparticles are arranged in chains comprising more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35 or 40 nanoparticles.

In another embodiment of the invention, the nanoparticles are arranged in chains, which have: i) a length less than 2.1010, 2.105, 2.103 or 2.102 nm, or ii) a number of nanoparticles in each chain less than 2, 5, 10, 102 or 103. In certain cases, short chains of nanoparticles may be desired or obtained, for example to promote the internalization of nanoparticles in cells or after partial or total destruction of the long chains.

In another embodiment of the invention, the nanoparticles are arranged in chains, which have: i) a length greater than 10-1, 1, 5, 10, 2.102, 2.103 or 2.105, or ii) a number of nanoparticles in every string greater than 2, 5, 10, 102 or 103. In some cases, long chains of nanoparticles may be desired or obtained to increase the amount of heat or compounds dissociated from the nanoparticles under the application of an acoustic wave or radiation or to prevent aggregation of the nanoparticles or allow uniform distribution of nanoparticles.

In another embodiment of the invention, the nanoparticles are arranged in chains, which have: i) a length between 10-1 and 1010 nm, or between 1 and 105 nm, or ii) a number of nanoparticles in each string between 2 and 105, 2 and 103, 2 and 102, or 2 and 50.

In another embodiment of the invention, the nanoparticles are arranged in chains when they are linked or bonded to each other or when the crystallographic directions of two adjacent nanoparticles in the chain are aligned, wherein such alignment is preferably characterized by an angle between two crystallographic directions belonging to two adjacent nanoparticles in the chains less than 90, 80, 70, 60, 50, 20, 10, 3 or 2 (degree).

Preferably, when the nanoparticles are synthesized biologically, the nanoparticles can be arranged in a chain: i) inside the organism which synthesizes them, also referred to as a living synthesizing organism, or ii) outside this organism. Preferably, the nanoparticles are arranged in a chain after or before their extraction or their isolation from this organism.

In one embodiment of the invention, the nanoparticles are not arranged in chains.

In another embodiment of the invention, the nanoparticles are synthesized chemically or are not synthesized by a living organism when less than 1, 2, 5, 10 or 100 step(s) of their production, such as the crystallization of iron oxide, the stabilization of iron oxide mineral, the organization of nanoparticles, involves or is due to a living organism. In some cases, a chemical synthesis may be defined as a synthesis involving a majority of steps, or more than 1, 2, 5 or 10 steps, or more than 1, 2, 5, 25, 50, 75 or 90% of the steps. stages, which involve chemical reactions occurring without the involvement of living organisms, or parts of living organisms such as DNA, RNA, proteins, enzymes, lipids.

In another embodiment of the invention, chemical synthesis may be used to produce a chemical substance or compound that mimics, copies, or reproduces the compartment, organelle, or other biological material, such chemical synthesis or chemical substance. that can be used or can lead to the production of nanoparticles. In some cases, the compartment, organelle or other biological material may be a lysosome, an endosome, a vesicle, preferably a biological material which has the capacity or function either to dissolve or transform crystallized iron into free iron, or to transform the free iron into crystallized iron. In certain cases, this transformation is partial and results preferentially in the destruction or in the formation of a partially crystallized assembly of iron atoms or ions, or preferentially results in a mixture of crystallized iron and non-crystallized iron. In certain cases, crystallized iron can be defined as an assembly of iron atoms or ions which leads to the presence of crystallographic planes, preferentially observable using a technique such as transmission or electron microscopy. scanning as a characterization method, and free iron can be defined preferentially as a set of iron atoms or ions which does not lead to the presence of crystallographic planes, highlighted preferentially by the absence of diffraction patterns, using for example transmission or scanning electron microscopy as a characterization method.

The invention also relates to nanoparticles for use, in which the nanoparticles are or are assimilated to chemical analogues of magnetosomes, such as iron oxide nanoparticles designated as Sigma nanoparticles (ref: 637106-25G), SPION20 (nanomag® -D-spio 20, Ref: 79-02-201), SPION50 (synomag-D50, Ref: 104-000-501), SPION100 (nanomag®-D-spio 100, Ref: 79-00-102) or nanoparticles synthesized using a method similar to that of these nanoparticles but having improved or additional properties such as a chain arrangement.

In some cases, chemical analogues of magnetosomes can be synthesized chemically and/or are not synthesized by magnetotactic bacteria.

In some cases, the chemical analogues of magnetosomes possess at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 properties in common with magnetosomes, these common properties being preferably ferrimagnetic behavior, preferably ferrimagnetic behavior. coercivity greater than 10-50, 10-10, 10-2, 1, 5, 10 or 100 Oe at a temperature preferably greater than 0, 5, 10, 50, 100, 200, 300, 500 or 1000 K, a large size, preferably a size greater than 1, 5, 10, 20, 50 or 70 nm, and/or a chain arrangement, preferably an arrangement of more than 1, 2, 5 or 10 nanoparticles in a chain.

In one embodiment of the invention, the nanoparticles or magnetosomes are purified to remove more than 10, 50 or 90 percent or mass percent of endotoxins and/or other biological material such as proteins or lipids originating from the living synthesizer organism or from magnetotactic bacteria. In certain other cases, the nanoparticles or magnetosomes are purified to remove less than 100, 99.9, 99, 95 or 90 percent or mass percent of endotoxins and/or other biological material. This purification step makes it possible to preferentially obtain purified nanoparticles or magnetosomes. In some cases, this percentage may be equal to [QBP -QAP] /QBP or QAP /QBP, where QBP and QAP are the quantities of endotoxins, biological material, proteins or lipids before and after the purification step , respectively.

In some cases, the purification step may consist of using a method or detergent(s) such as NaOH and/or KOH, which is (are) preferentially mixed with the living organism synthesizing or magnetotactic bacteria or bacterial debris, preferably to eliminate the organic matter or separate the organic matter from the inorganic material included in the nanoparticles or magnetosomes and preferably then be able to harvest the nanoparticle or the mineral magnetosome, preferably included in the nanoparticles or magnetosomes.

In some cases, the purified nanoparticles or magnetosomes are nanoparticle or magnetosome minerals.

In one embodiment of the invention, the nanoparticles according to the invention are medicines, medical devices, cosmetic products, biological products, products used for research purposes, or products used to determine the properties of biological samples.

The invention relates to a method of storing the composition according to the invention, which comprises at least one of the following steps:

- Step 1: Choose or prepare the composition in the form of a liquid suspension,

- step 2: Lyophilization, desiccation, dehydration of the composition, or elimination of water from the composition,

- Step 3: Storage of the composition in powder form, preferably for more than 1 second, 3 months or 1 year.
And

- Step 4: Suspending or resuspension of the composition, preferably in water.

EXAMPLE 1: Magnetosome minerals coated with citric acid and carboxy-methyl-dextran lyophilized in the presence of cryoprotectants for long-term storage and sustained anti-tumor activity.

We report a method to formulate natural iron oxide nanoparticles, called magnetosomes, by amplifying magnetotactic bacteria in non-toxic growth media, extracting these nanoparticles from magnetotactic bacteria under alkaline lysis, purifying them by heat above 400°C. Pure, non-pyrogenic, M-uncoated mineral magnetosomes are thus obtained, which are then coated with biocompatible citric acid or carboxy-methyl-dextran compounds, to give stable M-CA and M-CMD. The final steps of the formulation consist of adding sorbitol, preferably 5% sorbitol, to M-CMD and a mixture of sucrose and PEG 4000, preferably 3.75% sucrose and 1.25% PEG 4000, to M -CA, to lyophilize these mixtures, to obtain NP powders of (M-CMD) _f and (M-CA) _f which have long-term stability, preferably over at least 6 months, having preserved their properties before freeze-drying, i.e. their stability in suspension, their chain arrangement, their carbon content, as well as their surface charge and surface chemical groups. Furthermore, we established that (M-CMD) _f and (M-CA) _f are isotonic, preferably with osmolalities between 275 and 290 mosm/kg H ₂ O when reconstituting NPs in water, fully biocompatible, that is to say sterile, apyrogenic, non-cytotoxic towards healthy 3T3, L929 and V79 cells up to an NP concentration of 1 mg/ml, and effective in destroying PC3-luc prostate tumor cells when They are heated to a maximum temperature of 46°C for 30 minutes in the presence of these cells under the application of low intensity ultrasound or an alternating magnetic field. These results demonstrate that (M-CMD) _f and (M-CA) _f exhibit long-term storage capacity, full biocompatibility, as well as the potential to destroy tumor cells under hyperthermia.

Among the different types of nanoparticles (NPs), those based on iron oxide (IONPs) exhibit one of the highest levels of biocompatibility, thus enabling their use for medical applications, particularly cancer treatment. Chemically synthesized IONPs often suffer from a series of drawbacks such as the use of toxic compounds in their manufacturing process, crystallinity, non-uniform shapes, and/or small sizes with low magnetization. To overcome these drawbacks, a biological synthesis pathway has been developed, in which IONPs, called magnetosomes, are produced intracellularly by gram-negative magnetotactic bacteria (MTB). MTBs have adjusted the properties of magnetosomes over millions of years through a Darwinian process to result in an optimized system of magnetic guidance, also called magnetotaxis, in which the magnetic moment of the magnetosome aligns parallel to the Earth's magnetic field, allowing MTBs to move in this direction, thus facilitating their search for an optimal living environment. The magnetosomes are therefore made up of well-crystallized cuboctahedral nanocrystals of magnetite (Fe ₃ O ₄ ) surrounded by a stabilizing phospholipid bilayer originating from MTB. At the individual level, they have a size of 35 to 120 nm, which gives them ferrimagnetic properties. On the larger scale of magnetosome assembly, they are observed to form chains, which prevents them from agglomeration. While these characteristics led to greater antitumor efficacy for magnetosomes than for their chemical counterparts, these results were obtained with magnetosomes in unfinalized formulations.

Part of our work involves improving the magnetosome manufacturing process to make these NPs injectable in humans. To do this, toxic CMR or animal-derived compounds are first removed from MTB growth media. Next, the natural organic membrane of the magnetosomes which contains lipopolysaccharides known as endotoxins is removed and further processing is undertaken to produce mineral magnetosomes called M-uncoated, which are non-pyrogenic but prone to agglomeration/sedimentation due to strong magnetic dipolar interactions. Third, the uncoated M-s are coated with biocompatible compounds through coordination bonds between a coating material, i.e. citric acid (CA) or carboxy-methyl dextran (CMD), and iron cations on the surface of the mineral, to give stabilized M-CA or M-CMD. The following steps, which are the subject of this example, consist of adding cryoprotectants to M-CA and M-CMD and lyophilizing the mixtures obtained to transform them into an NP powder, called (M-CA) _f and ( M-CMD) _f , which can be stored for a long time without losing its properties.

To improve long-term storage stability and avoid disassembling the magnetosome mineral coating, it is preferable to store the coated magnetosome minerals as a powder rather than a liquid suspension. To remove water from M-CA and M-CMD, we lyophilized these NPs in the presence of different cryoprotectants, whose cryoprotective action probably relies on the creation of an amorphous glassy matrix that immobilizes the nanoparticles upon freezing and thus prevents the crystallized ice produced during this step from damaging the coating. As the effectiveness of cryoprotection likely depends on the type and concentration of the cryoprotectant used in a formulation, we tested different cryoprotectants, namely glucose , mannitol, PEG 4000, sorbitol, sucrose, trehalose, at different concentrations, which we added to M-CA and M-CMD. We then determined the optimized conditions making it possible to obtain stable and isotonic products (M-CA) _f and (M-CMD) _f , which can be stored in powder form and resuspended in water, while retaining their properties. physicochemical, i.e. the thickness/composition of their coating or the arrangement of the magnetosome chain, their biocompatibility, i.e. their sterility, their absence of pyrosis and their resistance to corrosion, that is to say their sterility, their non-pyrogenicity, their non-cytotoxicity towards healthy cells, as well as their ability to destroy PC3-luc tumor cells under conditions of hyperthermia mile of 41-46°C triggered by the application of low intensity ultrasound or alternating magnetic field.

MATERIAL AND METHODS:

Material : Magnetospirillum gryphiswaldense strain MSR-1 (DSM 6361) was purchased from the Deutsche sammlung von mikro -agencen und zellkulturen (Brunswick, Germany). The Pc3-luc tumor cell line derived from human prostate adenocarcinoma PC3 was purchased from Perkinelmer. The 3T3 mouse fibroblast cell line was purchased from the American type culture collection (ATCC® CCL-163™). Chinese male hamster lung cell line V79-4 was purchased from ATCC® (CCL-93). Mouse fibroblast cell line L929 was purchased from ATCC® (CCL-1). Potassium hydroxide (KOH) and citric acid were purchased in pharmaceutical grade from Merck. Carboxymethyl dextran (CMD) was purchased from TDB Labs. Sucrose, glucose, trehalose, mannitol, sorbitol, and polyethylene glycol 4000 (peg 4000) were purchased in pharmaceutical grade from Merck. Dextran T1 was purchased from pharmacosmos. Tryptic soy broth (TSB) and thioglycolate fluid medium (FTM) were purchased from Merck. Phosphate-buffered saline (PBS), sodium hydroxide (NOH), hydrochloric acid (HCl), nitric acid (HNO ₃ ), and Triton X-100 were purchased from Thermo fisher scientific. Dulbecco's Modified Eagle's Medium (DMEM) with/without phenol red, fetal bovine serum (FBS), penicillin-streptomycin, HEPES buffer solution, 0.25% trypsin solution - EDTA, and trypan blue solution were purchased from Gibco. Resazurin dye was purchased from Invitrogen.

Culture of MSR-1 magnetotactic bacteria: Briefly, MSR-1 bacteria were first amplified in two pre-culture (PC) steps with the iron-deficient pre-culture medium, and then cultured in the last step with the appropriate conditions to promote the synthesis of magnetosomes. During PC 1, 300 µl of MSR-1 cryo-stock was incubated in 50 ml of pre-culture medium at 29.5°C for 6 days in a static state followed by 1 day under shaking at 110 rpm. Then, PC2 was performed by transferring all cells into a 15 L bioreactor (Applikon) containing 6 L of pre-growth medium and stirred at 200 rpm and 29.5°C for 3 days. During the last culture step, PC 2 cells were transferred to a 40 µl bioreactor (Applikon) filled with 26 µl of growth medium. The culture conditions such as oxygen concentration, temperature and stirring speed were respectively maintained between 0.1 and 1%, at 29.5°C and under 200 rpm throughout the fermentation. the pH was maintained at 6.9 by the automatic addition of an acidic medium (pH = 3), called a “ fed -batch” solution. Bacterial growth was monitored every 24 hours by taking a 5 ml sample of the culture to determine the optical density at 565 nm (OD ₅₆₅ ) using a UV-visible spectrophotometer. MSR-1 magnetotactic bacteria (MTB) were cultured for a total of 19 days, using bacterial growth media composed of pharmaceutical grade chemicals, free of toxic compounds and heavy metals other than iron. The pre-growth medium (PGM), growth medium (GM) and fed-batch medium (FBM) include: i) sodium lactate (preferably 2.6 g/L in PGM, 1.3 g/L in L in GM, 100 g/L in FBM), ii) ammonium chloride (preferably 0.4 g/L in PGM, 0.2 g/L in GM, 4.8 g/L in FBM), iii) magnesium sulfate heptahydrate (preferably 0.1 g/L in PGM, 0.03 g/L in GM, 2.4 g/L in FBM), dipotassium phosphate (preferably 0.5 g/L in PGM, 0.07 g/L in GM, 6 g/L in FBM), iron(III) chloride hexahydrate (preferably 0 g/L in PGM, 0 g/L in GM, 2 g/L in FBM), elixir mineral (preferably 0.5 ml in PGM, 0.08 ml in FBM, 7 ml in FBM), vitamin elixir (preferably 0.1 ml in PGM, 0.07 ml in GM, 1 ml in FBM). The mineral elixir includes 1 g/L of iron (II) sulfate heptahydrate and 30 g/L of calcium chloride. The vitamin mixture includes 0.002 g/L biotin, 0.4 g/L calcium penthotenate, 0.002 g/L folic acid, 2 g/L inositol, 0.4 g/L nicotinic acid, 0.2 g/L p-aminobezoic acid, 0.4 g/L pyridoxine HCl, 0.2 g/L riboflavin, 0.4 g/L Thiamine HCl. The first 10 days include 2 pre-culture steps used to amplify the bacteria in the pre-culture medium lacking the iron source. The third stage lasts approximately 9 days. It consists of promoting the intracellular synthesis of magnetosomes by cultivating MSR-1 bacteria in their optimized microaerophilic conditions and gradually adding sources of iron ions via a fed-batch solution. At the end of the bacterial culture, 32 l of bacterial culture was concentrated to reduce the liquid volume using a tangential filtration column. Then, the MSR-1 concentrate (approximately 5 L in total) was stored at -80°C for the following steps of magnetosome production.

Extraction and purification of magnetosomes: Briefly, concentrated MSR-1 cells were diluted to an OD ₅₆₅ value of 20 and then lysed in 2 m KOH at 80°C for 1 h under mechanical shaking at 150 rpm. Using magnetic selection, magnetosome chains (MgC) were collected and washed 2 times with 1x phosphate-buffered saline (PBS) followed by 3 times washing with deionized water. After being concentrated in 50 ml conical tubes, the MgCs were frozen at -80°C for 24 h and then dried at -50°C, 0.003 mbar for 24 h using a freeze dryer (labconco, free zone 70020 2.5 l) to transform them into powder. Then, 100 mg of this powder was heated at 6°C/min in a muffle furnace using a heating program comprising several heating stages at temperatures between 50°C and 420°C (maintained for 2 h). , to purify organic matter from post-extracted magnetosomes and obtain magnetosome minerals or “uncoated magnetosomes” (M-uncoated).

Coating of M- Uncoated with Citric Acid (CA) and Carboxymethyl-Dextran (CMD): The coating process was carried out under aseptic conditions, using a biosafety hood and non-pyrogenic materials. CA and CMD solutions were prepared in pyrogen-free water at 25 mg/ml and 150 mg/ml, respectively. These solutions were then filtered with a 0.22 µm polyethersulfone filter to be sterilized. Then, 200 mL of each solution (CA or CMD) was added to a 1 L glass beaker containing 2 g of M-uncoated (corresponding to 1 g of Fe) and 300 mL of pyrogen-free water. Each suspension was then sonicated for 1 min in pulsed mode with a pulse length of 0.1 s and a pulse interval of 0.1 s, at 20 W and room temperature, using a probe sonifier (PS) (branson, digital sonifier s-250d) with 25 mm diameter tips. After the short-time sonication, the pH values were adjusted to 6 and 4.5 for CA and CMD, respectively, using 1 M NaOH or 1 M Hcl. Then, these two suspensions were sonified at again for 1 h using PS (Branson, s-250d digital sonifier) with the same parameters as previously described to prepare mineral magnetosomes coated with CA and CMD, called M-CA and M-CMD respectively. After sonication, the M-CA and M-CMD suspensions were centrifuged at 10°C and 3380 g for 45 min to remove excess coating agents in the supernatant. Each magnetosome-coated mineral was then resuspended in 10 ml of water to obtain a final concentration of 100 mg/ml iron and stored at +4°C.

Selection of cryoprotectants:Under aseptic conditions, 0.5 ml of M-CMD suspension at 100 mg/ml iron was mixed with 0.5 ml of different solutions of cryoprotectants, such as glucose, mannitol, sorbitol, sucrose and trehalose, to obtain samples formulated from 1 ml to 50 mg/ml of iron containing 5% or 10% w/v of cryoprotectant. Each sample was immediately frozen in liquid nitrogen for 15 min after 30 s vortexing, then freeze-dried at -50°C (storage temperature), 0.003 mbar for 20 h for primary desiccation (labconco, free zone 70020 2, 5l). Then, secondary desiccation was triggered for 6 hours by increasing the temperature to 40°C (storage temperature) with a heating rate of 0.3°C/min while maintaining the pressure at 0.003 mbar. Likewise, the suspension of M-CA at 100 mg/ml of iron was tested with different solutions of cryoprotectants to obtain samples formulated from 1 ml to 50 mg/ml of iron containing a cryoprotectant at 5% or 10% w/ v for glucose, mannitol, sorbitol, trehalose and sucrose; 0.5%, 2.5%, 5%, 7.5%, 10% and 15% w/v of cryoprotectant for peg 4000 and dextran T1; and 2.5%, 3.75%, 5%, 6.25% and 7.5% w/v of cryoprotectant for the combinations sucrose - peg 4000 and sucrose - dextran T1 with mass ratios sucrose - PEG or dextran of 1:1, 1:2, 1:3, 2:1, 2:2, 2:3, 3:1, 3:2 and 3:3. After freeze-drying, some formulated magnetosome samples were resuspended in 1 ml of sterile water for further characterizations. The selected formulations of M-CA and M-CMD with cryoprotectants were called (M-CA)_f and (M-CMD)_f, respectively.

Transmission electron microscopy (TEM): 1 ml of MSR-1 bacteria at the end of culture was diluted to an OD _{565 nm} value of 1 then rinsed twice with deionized water by centrifugation at 2400 g for 10 min. The different suspensions of magnetosomes were diluted to 50 μg/ml of iron. Then, 7 µl of each sample was deposited on a carbon-coated copper grid (300 mesh from Oxford Instruments). The grid was then left dry at room temperature for at least 3 h before being observed under a transmission electron microscope (Jeol JEM-2100) operating at 200 kv. Using image j software, nanoparticle size was estimated by measuring the diameter of approximately 400 randomly chosen mineral cores from the magnetosome chains inside MSR-1 bacteria.

Colloidal stability by colorimetric assay: 1 ml of magnetosome suspension was vortexed for 30 s then a 50 μl sample was immediately taken, called the “t ₀ sample”. The suspension was then left on the bench for 2 hours and another 50 µl sample was then taken from the upper part of the liquid, called the “t _2h sample”. For the transformation into iron ions, each sample was dissolved overnight in 950 μl of 37% v/v HCl at room temperature. Then 20 µl were taken and mixed with 50 µl of H ₂ O ₂ at 20% v/v for 15 min to oxidize Fe ^{2 +} into Fe ³⁺ ions. After that, the solution was added with 880 µl of ultrapure water and then mixed with 50 µl of 2 m KSCN to form the complex between Fe ³⁺ ions and thiocyanate ions, whose absorption at 476 nm was measured in a spectrophotometer. The iron concentration was then determined using a calibration range. The stability rate of the magnetosome suspension was calculated as the ratio between the iron concentration of sample t _2h and that of sample t ₀ .

Osmolality measurement: 30 µl of magnetosome suspension with 50 mg/ml iron was introduced into a micro-sample tube and then placed on the osmometer plate (advanced osmometer, model 2020) to determine osmolality of each sample.

CHNS elemental analysis: 5 mg of magnetosome powder was packed in an aluminum capsule and introduced into the CHNS elemental analyzer (thermofisher, flash 2000) to determine the level of carbon and nitrogen from organic materials in each preparation.

Fourier transform infrared spectroscopy (FT-IR): 2 mg of magnetosome powder was deposited on a germanium crystal plate, connected to an FT-IR spectrometer. The tip was lowered until it contacted the crystal and therefore the powder sample. The FT-IR spectra were then recorded in ^a scanning range of 590 - 4000 cm ^-1 with a resolution of 4 cm ^-1 .

Zeta potential measurement: Each magnetosome suspension was diluted to 50 µg/ml in iron with ultrapure water. It was then divided into five 2 ml samples to be prepared at different pH varying from 2 to 10 to 25°C using HCl and NaOH solutions. 1.5 ml of each sample was then added to a 4.5 ml disposable cuvette and measured in a zeta potential assay.

Residual moisture analysis: 10 mg of freeze-dried powder was placed in an aluminum oxide crucible and introduced into the thermogravimetric analysis (TGA) instrument. Then, under a flow of nitrogen gas, the sample was heated from 30°C to 120°C at 6°C/min and kept at 120°C for 1 h to determine the weight loss of the freeze-dried powder.

Sterility test: Under aseptic conditions, 20 mg of each magnetosome powder was incubated in two conditions, 10 ml of tryptic soy broth (TSB) at 25°C and 10 ml of fluid thioglycolate medium (FTM) at 35°C. c for 14 days. Culture media without NP were considered negative controls. On the last day, 1 ml was taken from each sample and placed against the magnet for 10 min to collect only the medium. Turbidity was then measured with 900 μl of this medium at 600 nm. And the remaining 100 µl were incubated on a solid Luria-Bertani agar plate at 25°C or 35°C for 3 days to possibly detect contaminating colonies. The other tests, i.e. the growth promotion test and the method adequacy test, were also carried out in parallel with the magnetosome sterility test in order to verify the robustness of this test.

Quantification of endotoxin by the LAL test ( limulus amebocyte lysate ):Endotoxin quantification was carried out on different magnetosome powders using a “pierce chromogenic endotoxin quant” kit under aseptic conditions. All materials coming into contact with the NP samples were sterile and nonpyrogenic. Prior to testing, each magnetosome powder was prepared at 40 µg/ml iron in endotoxin-free water and then heated at 70°C for 15 min to denature any residual protein interfering with the assay. After that, 25 µl of this suspension was introduced into a 96-well plate pre-equilibrated at 37°C for at least 10 minutes. Maintaining the plate at 37°C, 25 µl of the reconstituted amebocyte lysate reagent was added to each well and incubated for 12 minutes, followed by incubation with 50 µl of the reconstituted chromogenic substrate for 6 minutes. Finally, 25 μl of 25% acetic acid was added to each well to stop the reaction. The optical density of this mixture was then measured at 405 nm in a microplate spectrophotometer. The endotoxin concentration was determined by the endotoxin standard curve ofe. coliwhich was established at the same time as the samples.

Growth of different cell lines: 3T3, L929, V-79, PC3-Luc: A 2 ml cryotube of different cell lines (3T3, L929, V-79, or PC3-Luc) was thawed in a water bath at 37°c for 10 minutes. Each of these cell lines was then introduced into a 75 cm ² culture flask containing 10 ml of appropriate culture medium, that is to say DMEM supplemented with 10% v/v of fbs, 1% v/ v penicillin-streptomycin mixture used for 3T3, V-79, and PC3-Luc, while DMEM supplemented with 10% v/v hs, 1% v/v penicillin-streptomycin mixture, and 1% v/v of HEPES buffer for L929. After being placed in the flask, the cells were maintained at 37°C in a 5% CO ₂ incubator. The culture medium was renewed twice a week for each flask. When the cells reached approximately 80% confluence, cell passage was performed. For this, all liquid medium was first removed from the flask, and 1 ml of 0.25% trypsin-EDTA was added to collect all adherent cells in suspension. Then, only 0.5 ml of cell suspension was kept to be mixed with 10 ml of fresh medium in a new 75 cm ² culture flask and reincubated at 37°C in the 5% CO ₂ incubator. After 3 passages, each cell line was ready for further experiments.

Cytotoxicity test. The 3T3, L929 and V-79 cell lines were each seeded on a sterile 96-well plate (10 ⁴ cells/well), then incubated overnight at 37°C in a 5% CO ₂ incubator for cell adhesion. After that, all the medium was carefully removed from the plate. 100 µl of the lyophilized (M-CA) _f or (M-CMD) _f suspension, prepared at different concentrations in the appropriate culture medium without phenol red for each cell line, were then added to each well. In 3T3 cell plates, V-79, L929, 0.001, 0.1, 0.25, and 1 mg/ml iron magnetosome suspensions were added. After adding the nanoparticle suspensions, the plates were reincubated for 24 h at 37°C in a 5% CO ₂ incubator. To determine cell viability, 10 μl of resazurin reagent was added to each well and homogenized with shaking at 120 rpm for 10 min. The plates were then incubated at 37°C in the 5% CO ₂ incubator for approximately 4 h. Once the blue color of the untreated cell wells changed to pink, all liquid from each well was transferred to a 1.5 ml Eppendorf tube. The tube was then centrifuged at 14100 g for 10 min to remove any nanoparticles that could interfere with the assay. Then, 100 μl of the supernatant was transferred to a new 96-well plate to be measured at the excitation and emission wavelengths of 530 and 590 nm, respectively, using a fluorescence reader. Cell viability was calculated using the formula: Or , is the fluorescence intensity of the treated wells, is the fluorescence intensity of the blank measured in empty wells containing 100 µl of medium mixed with 10 µl of resazurin, and is the fluorescence intensity of control wells measured in untreated cells incubated in 100 µl of medium mixed with 10 µl of resazurin.

Hyperthermia treatments under AMF (alternating magnetic field) and LIU (low intensity ultrasound) applications. PC3-luc cells were seeded on a sterile 96-well plate (3x10 ⁴ cells/well) and incubated overnight at 37°C in a 5% CO ₂ incubator for cell adhesion. Then, all medium was removed from each well and adherent cells were rinsed 2 times with phenol red-free DMEM (white DMEM). 100 µl of suspension of (M-CA) _f or (M-CMD) _f prepared at 1 mg/ml iron in white DMEM supplemented with 10% v/v FBS, 1% v/v penicillin-streptomycin and 1% v/v HEPES buffer was added to each well. Then, all wells were reincubated for 3 hours at 37°C in a 5% CO ₂ incubator. After that, the hyperthermia experiment was divided into three conditions in which for each condition there were 3 wells containing only Pc3-luc cells, another 3 wells containing cells incubated with (M-CA) _f , and 3 other cell wells with (M-CMD) _f . In the first condition, the wells were not exposed to any source of hyperthermia. In the second condition, 3 wells per time were placed in a polystyrene holder positioned in the center of a copper coil, in which an AMF of 42 mt and 195 KHz was applied for 30 min with easy-to-use heat equipment. use to heat cells. in the third condition, each of the 3 wells was dipped in an adapter filled with degassed water to make the connection with a plane ultrasonic transducer (3.6 cm in diameter) and thus be exposed for 30 min to an ultrasonic intensity comprised between 0.3 and 1 w/cm ² , at 1 MHz and in continuous mode, produced by an ultrasound generator (Primo Therasonic 460). To limit evaporation of the culture medium, each well was covered with a cap when exposed to a source of hyperthermia. The temperature was monitored over time using a flexible thermocouple probe (physitemp, it-18) inserted in each well and Dasylab software. The sample SAR was calculated using the following equation: where Cmedium = 4.2 jg ^-1 .k ^-1 is the specific heat capacity of water, C _fe , which is measured in g of iron per g of water, represents the iron concentration in the magnetosome sample , andΔ t/ (measured in °c/s) is the slope of the temperature variation with time of the magnetosome sample after subtracting that of the control samples. After the hyperthermia treatments, all wells were incubated for 24 hours at 37°C in a 5% CO ₂ incubator. After that, cell viability in each well was determined using resazurin reagent.

Iron quantification of magnetosomes internalized by cells. After removing all the supernatant for the cell viability test, the PC3-luc cells adhering to the bottom of the wells were gently rinsed 3 times with 300 µl of 1x PBS. Each well was then incubated with 50 µl of 0.25% trypsin - EDTA for 1 min at 37°C and 5% CO ₂ , followed by the introduction of 150 µl of complete DMEM. After that, all the liquid from each well (200 μl) was transferred to a 1.5 ml Eppendorf tube. After a few seconds of vortexing, a 20 μl sample was taken into the Eppendorf tube and mixed with 20 μl of trypan blue to determine the cell concentration using a Malassez hemocytometer. 180 μl of the remaining cell suspension was centrifuged at 14,100 g for 20 min to remove all supernatant. The cell pellet was then dissolved overnight in 20 µl of 37% w/v HCl and 145 µl of 70% w/v HNO ₃ at room temperature. Finally, the solution was diluted with ultrapure water to a total volume of 5 mL and analyzed in an inductively coupled plasma mass spectrometer (agilent, 7900 icp-ms) to determine the iron content of the internalized magnetosome in PC3-luc cells.

Statistical analyses. Data were analyzed using Graphpad Prism version 8.0. All measurements were performed in triplicate (n = 3) and data were reported as mean standard deviation (sd). Statistical comparisons were performed using one-way anova and differences were considered significant when * p < 0.05, ** p < 0.01, and *** p < 0.001.

RESULTS AND DISCUSSIONS.

Magnetosome-based formulation: summary of previously published first steps and presentation of additional/complementary steps proposed in this study. This study presents additional aspects regarding the implementation of a magnetosome-based formulation for injection in humans. The first steps of this formulation are summarized. Step 1 involves culturing magnetotactic bacteria, leading to spiral-shaped magnetotactic bacteria containing magnetosome chains in their cytoplasm with average magnetosome sizes of 36.6 (standard deviation 6.4 nm). Step 2 includes extraction of magnetosomes from magnetotactic bacteria from step 1, using KOH bacterial lysis and magnetic separation of magnetosomes from bacterial debris, leading to extracted magnetosomes arranged in chains (Mg-Ch), where each magnetosome is surrounded by a bacterial inflammatory membrane, which seems too immunogenic in the absence of specific treatment for injection into humans. Step 3 includes purification of Mg-Ch by combustion to remove/denature bacterial organic matter from Mg-Ch, leading to non-pyrogenic (M-uncoated) magnetosome minerals with a low carbon percentage of 0.2 %, which tend to aggregate due to their bare surface. Uncoated M-s are coated with two biocompatible compounds, namely citric acid (CA) or carboxy-methyl dextran (CMD), resulting in reconstituted non-pyrogenic coated magnetosome minerals (M-CA and M-CMD) in chains.

The goal of this example is to add a few more steps in the preparation of coated magnetosomes so that M-CA and M-CMD can be preserved over a long period of time, while preventing degradation of the coating. To achieve this goal, M-CA and M-CMD were stored as anhydrous powder, which can be reconstituted into NP suspension at any time, as needed. We then verified that reconstituted M-CA and M-CMD, denoted by (M-CA) _f , (M-CMD) _f , (M-CA) _fw , (M-CMD) _fw for M-CA and M- Lyophilized CMDs before and after resuspension of NPs in water, respectively, retained their physicochemical properties and antitumor activity.

Formulation of M-CA and M-CMD in the presence of a cryoprotectant for long-term storage. Here we determine the conditions for manufacturing a formulation of injectable magnetosomes exhibiting long-term stability. The latter is evaluated by measuring the percentage of absorption stability, measured at 480 nm over time, of a suspension of magnetosomes, preferably 50 mg of magnetosomes for 2 hours after its homogenization. The percentages of 100% and 0% correspond to stable and unstable suspensions respectively. We chose to perform the stability measurements with 50 mg of magnetosomes because we plan to administer this amount of magnetosomes into a human prostate tumor of 1 to 2 cm in diameter based on our preclinical efficacy data in mice extrapolated to humans. Additionally, a time frame of 2 hours was chosen for stability assessment, which appears to be long enough for a nurse or doctor to deliver a stable magnetosome suspension into tumors. Since the purified magnetosomes devoid of active bacterial organic matter (M-uncoated) were unstable, i.e. their stability percentage was 0%, they were coated with CA and CMD, which gave M-CA and M-CMD, which are perfectly stable, that is to say their percentage of stability is 100%, which is higher than that of M-uncoated and M-gC. When M-CA and M-CMD are lyophilized in the absence of a specific compound capable of preserving their coating, they form NP powders designated as (M-CA) _fd and (M-CMD) _fd characterized by the presence of clustered magnetosomes. When (M-CA) _fd and (M-CMD) _fd are resuspended in water after the freeze-drying step, they sediment rapidly, resulting in a stability percentage of less than 5%. To allow the reconstitution of M-CA and M-CMD in stable aqueous suspension of NP after lyophilization, i.e. without damaging the coating M-CA/M-CMD, M-CA and M-CMD at the dose intended therapy of 50 mg/ml were mixed with different cryoprotectants, i.e. glucose, mannitol, sorbitol, sucrose or trehalose, used at mass percentages of 5% and 10%. After being freeze-dried, these formulations were resuspended in water to determine their colloidal stability. Considering M-CMD first, only sorbitols at 5% and 10% manage to maintain 100% colloidal stability of NPs. Furthermore, the osmolality of the M-CMD suspension increases with the percentage of sorbitol added to the M-CMD suspension, increasing from 0 mosm/kg H ₂ O in the absence of cryoprotectant to 280 and 560 mosm/kg H ₂ O for 5% and 10% sorbitol, respectively. M-CMD formulated with 5% sorbitol results in an osmolality value within the plasma osmolality range (275 - 290 mosm/kg H ₂ O), which is acceptable for human injection. Regarding M-CA, M-ca mixed with 10% glucose, sorbitol or sucrose leads to 100% colloidal stability. However, these formulations are hypertonic, that is to say their osmolality is greater than 300 mosm/kg H ₂ O. To solve this problem, we selected M-CA mixed with 10% sucrose which leads to a osmolality of 330 mosm/kg H ₂ O, which is the closest value to the plasma osmolality, and we added to this mixture either PEG 4000 or Dextran T1, to reduce the osmolality of the formulation to plasma osmolality. While the use of Dextran T1 helps stabilize the formulation, it gives an osmolality lower than plasma osmolality. Therefore, Dextran T1 was discontinued. On the other hand, by combining sucrose with PEG 4000, a 100% stable formulation was achieved while the sole addition of 7.5% sucrose or 15% PEG 4000 led to a stability of approximately 90 or 50%, respectively. This result suggests a synergistic effect between sucrose and PEG 4000, which appear to support each other, i.e. PEG 4000 could effectively protect the NPs during freezing by encapsulating the NPs in a glassy matrix, and the Amorphous sucrose could serve as a “water replacement” reservoir allowing the formation of hydrogen bonds with citric acid on the NP surface, and consequently preserving the integrity of the coating against dehydration-induced stress. The optimal combination of sucrose and PEG 4000 corresponds to 1.25% PEG 4000, i.e. the lowest concentration of PEG 4000 to prevent the risk of molecular mobility and degradation of the formulation during storage, which could be a consequence of too high a concentration of PEG, and the largest percentage by mass of sucrose, that is to say 3.75%, to obtain a stable formulation of M-CA in suspension. The M-CA formulation with 1.25% PEG 400 and 3.75% sucrose has an osmolality of 225 mOsm/Kg H ₂ O, which can be adjusted to be isotonic upon reconstitution.

M-CA/M-CMD retain their physicochemical properties after their formulation.To examine whether magnetosomes formulated with 5% sorbitol for M-CMD, or 1.25% PEG 400 and 3.75% sucrose for M-CA, which are designated as (M-CMD)_fand (M-CA)_fbefore removal of cryoprotectant and as (M-CMD)_wfand (M-CA)_wfafter the removal of the cryoprotectant, retained their physicochemical properties, we compared the percentages of carbon and nitrogen as well as the FT-IR spectra and the surface charges of the formulated magnetosomes with those of the magnetosomes harvested at the different stages preceding the formulation,i.e. M-CA, M-CA, M-CA and M-CA.i.e.M-CA, M-CMD, M-gC, and M-uncoated. Regarding the mass percentages of carbon and nitrogen in the magnetosomes at the different formulation stages, they first decrease in the percentages of carbon and nitrogen of %C=22.7% and %N=2, 2% in M-gC up to %C=012% and %N=0.01% in M-uncoated, corresponding to the removal of the magnetosome membrane in M-gC to give M-uncoated. Then, they increase a first time from M-uncoated to M-CA (%C =1.61%, %N=0.01%) and M-CMD (%C=3.80%, %N=0 .01%), which is due to the addition of a coating of CA and CMD to the surface of M-uncoated, and a second time of M-CA and M-CMD to (M-CA)_f(%C=18.60% and %N=0.01%) and (M-CMD)_f(%C=18.56% and %N=0.01%), when the cryoprotectant is added to the formulated magnetosomes. In the final step, these percentages decrease again after washing the cryoprotectant from the formulated magnetosomes, i.e.that is to sayup to %C=1.6% and %N=0.01% in (M-CA)_fwand %C=3.83%, %N=0.01% in (M-CMD)_fw. The FT-IR spectra of magnetosomes at different stages of their formulation also confirm the CHNS trends described above. Indeed, the FT-IR spectrum of Mg-C presents P-O vibration bands (at 1037 cm^{- 1}), C-W (at 1410 cm^-1), N-H (at 1542 and 1641 cm^-1), and O-H (at 3276 cm^-1), corresponding to the functional groups present in the phospholipid membrane of the magnetosome. Concerning M-uncoated, its FT-IR spectrum does not indicate the presence of peaks above 1100 cm^{- 1}, which is consistent with the removal of most organic matter in this sample. The two peaks at 612 and 693 cm^{- 1}are attributed to Fe-O stretching vibrations, which originate from the iron oxide included in the mineral core of the magnetosome, and are therefore present in all FT-IR spectra of the different types of magnetosomes. Coating of M-uncoated with CA and CMD leads to a series of peaks in the range of 1000 - 1300 cm^-1and 1630 - 1750 cm^{- 1}, which correspond to the stretching vibrations of c-o and c=o, respectively, attributed to the carboxylate groups of CA and CMD in M-CA and M-CMD. Further in the formulation process, lyophilized magnetosomes (M-CA)_fand (M-CMD)_fdisplay intense bands of alcohol and alkane groups,eg.C-O, O-H and C-H stretching vibration bands in the range of 970 - 1250 cm^-1, 3200 - 3550 cm^-1and 2850 - 3000 cm^-1, respectively, corresponding to sucrose and PEG 4000 in (M-CA)_f, and sorbitol in (M-CMD)_f. Finally, after a washing step, (M-CA)_fwand (M-CMD)_fwpresent FT-IR spectra very close to those of M-CA and M-CMD, indicating that the washing procedure easily separated the cryoprotectant from the coated magnetosome, due to the lack of strong bonding between the cryoprotectant molecules and magnetosome coating agents. CHNS and FT-IR measurements suggest that the surface of the formulated magnetosomes remains unchanged compared to that of the magnetosomes coated before formulation. To confirm this deduction, we measured the surface charges of formulated magnetosomes which we compared to those of unformulated magnetosomes. (M-CA)_fand (M-CMD)_fexhibit surface charges very similar to those of M-CA and M-CMD when the pH of the suspensions containing these NPs varies between 2 and 10. This behavior indicates that the cryoprotectant effectively maintains the surface charges of magnetosomes in (M-CA )_fand (M-CMD)_f, probably by effectively protecting the coating layers in M-CA and M-CMD from degradation/removal during freeze-drying, and preventing the cryoprotectant from strongly binding to the NPs. Furthermore, under the conditions of use for human injection, itthat is to sayat the therapeutic dose of 50 mg/ml in iron and at a pH of  6.5, (M-CA)_fand (M-CMD)_fpresent surface charges less than -30 mv, creating the conditions for strong repulsive electrostatic forces in suspensions of (M-CA)_f/ (M-CMD)_f, which can prevent the agglomeration of NPs caused by magnetic dipolar interactions and stabilize these NPs in suspension. Electron microscopy measurements carried out on formulated magnetosomes washed of their cryoprotectant,that is to sayon (M-CA)_fwand (M-CMD)_fw, show that the formulated magnetosomes maintain a chain arrangement after the lyophilization step, a property complementary to that of their surface charge, ensuring their stability and preventing their aggregation. Finally, the long-term stability of the formulated magnetosomes, which is sought here, is ensured when the quantity of water remaining in the lyophilized product after the lyophilization process, called residual moisture (MR), is sufficiently low, c'that is to saytypically less than 3% for a pharmaceutical product. The RM content of (M-CA)_fand (M-CMD)_fwas measured by introducing these NPs into a TGA, heating them at 120°c for 1 h with a heating rate of 6°c/min, and measuring the weight loss of these NPs attributed to the evaporation of the 'water. RM contents of (M-CA)_f and (M-CMD)_fare in the range of (1.8-2.4)%, which is pharmaceutically acceptable. In addition, thanks to this efficient freeze-drying process, (M-CA)_fand (M-CMD)_fcan be resuspended in water, preferably after 6 months, where they retain their stability after a six month storage period.

Sterility/non- pyrogenicity of formulated magnetosomes. Compared to unformulated magnetosomes, formulated magnetosomes must not only preserve their physicochemical properties, but also their biocompatibility. To study the latter aspect, we first examined the sterility of magnetosomes formulated by introducing (M-CA) _f and (M-CMD) _f for 14 days in tryptic soy broth (TSB) and fluid medium solutions. of thioglycolate (FTM) at 30°C and 37°C, respectively. Indeed, these conditions are known to amplify bacterial contaminants when they are initially present in a tested sample, and therefore to enable their detection. To determine the presence (or absence) of bacteria in (M-CA) _f and (M-CMD) _f , the optical density of these suspended NPs was measured at 600 nm (OD ₆₀₀ ) at the end of the time incubation. The OD ₆₀₀ value reflects the turbidity of these suspensions, which is associated with the presence of a potential bacterial contaminant. (M-CA) _f and (M-CMD) _f exhibit a low OD ₆₀₀ < 0.1, which is comparable to the OD ₆₀₀ < 0.1 of M-uncoated and sterile TSB and FTM media before incubation (NC), indicating the absence of bacterial contaminants in (M-CA) _f and (M-CMD) _f . This result also confirms the absence of colonies in the agar plates inoculated with (M-CA) _f and (M-CMD) _f . To complete the evaluation of the sterility and non-pyrogenicity of (M-CA) _f and (M-CMD) _f , the endotoxin concentration of these NPs was measured. (M-CA) _f and (M-CMD) _f appear to contain a very low endotoxin concentration of 2-5 EU/mg iron, comparable to the values measured for uncoated M-s. Overall, these results highlight the effectiveness of the magnetosome purification step to remove bacterial contaminants from M-gC, whose initial presence before specific treatment is revealed by the high OD ₆₀₀ value of M- gC of 1-2.5 as well as by a high endotoxin concentration of M-gC of 50 EU/mg in iron. Such behavior can be attributed on the one hand to a two-step depyrogenation process, i.e. first mixing the M-gC with KOH at 80 °C and then heating the M-gC above 400°C, and on the other hand at formulation steps carried out under ascetic conditions, that is to say the coating of magnetosome minerals with CA or CMD, the addition of cryoprotectant with M-CA and M-CMD coated magnetosomes, the lyophilization of the mixture obtained is carried out under a sterile hood.

Non-cytotoxicity of formulated magnetosomes. To examine the cytotoxicity of formulated lyophilized magnetosomes, 100 µl of suspensions (M-CA) _f and (M-CMD) _f with concentrations varying between 0.001 and 1 mg of iron NP per ml were brought into contact with different cell lines of mammals, namely 3T3, L929 and V-79 cells, for 24 hours at 37°c. After this treatment, cell viability was assessed by measuring cellular metabolic activity using the resazurin test. All cell lines displayed viability greater than 70% for all tested concentrations of (M-CA) _fw and (M-CMD) _fw , revealing that these NPs are not cytotoxic under the conditions tested, and that the presence of a cryoprotectant and a lyophilization step in the magnetosome formulation does not result in additional cytotoxicity.

Effective treatment of hyperthermia using mineral magnetosomes formulated and excited under an alternating magnetic field and ultrasonic sources . We investigate whether (M-CA) _f and (M-CMD) _f retain the therapeutic activity observed in unformulated M-CA and M-CMD magnetosomes. Specifically, we examine whether (M-CA) _f and (M-CMD) _f effectively kill prostate tumor cells when 100 µl of (M-CA) _f or (M-CMD) _f at a concentration of 1 mg of np in iron per ml of aqueous suspension are incubated with PC3-luc cells for 3 h, and the resulting mixture is exposed for 30 minutes to AMF of 42 mt and 195 KHz or low intensity ultrasound of 0, 3-1 w/cm ² and 1 MHz. Such excitation conditions lead to an increase in the temperature of these mixtures from ambient temperature of 22-25°C to a moderate hyperthermia temperature of 46°C, where the temperature increase is significantly faster with ultrasound than with magnetic excitation, leading to a maintenance of the temperature at 46°C for almost 30 minutes and just under 10 minutes with ultrasound and magnetic field, respectively. The specific absorption rates (SAR) of (M-CA) _f and (M-CMD) _f are measured upon exposure of these NPs to AMF/LIU after subtracting the initial temperature increase due to the AMF/LIU in the absence of NP. While (M-CA) _f and (M-CMD) _f display a large SAR value of 229 ± 16 W/g _Fe under AMF excitation, they produce a near-zero SAR value under LIU, highlighting the nature different from heating with the two excitation sources, that is to say heating coming from the excitation of the NPs by the AMF or from the absorption of the ultrasound energy by the cells, using the application AMF or LIU, respectively. Moreover, the heating properties of the two types of formulated magnetosomes, i.e. (M-CA) _f and (M-CMD) _f are similar. This indicates that the differences between the two coatings in terms of nature/composition, coating thickness, carbon content, do not significantly affect the heating properties of (M-CA) _f and (M-CMD ) _f . We can conclude that some heating contributions, such as the Brownian contribution in the induction mechanism, which may depend on the coating properties, are not dominantly involved in the observed heating properties. To more precisely evaluate the effectiveness of heat treatment in killing tumor cells, Pc3-luc cells treated as described above were reincubated overnight at 37°C under 5% CO ₂ . Their viability was then measured. In the absence of heat treatment, PC3-Luc cells incubated with (M-CA) _f and (M-CMD) _f exhibit a viability of  80%, indicating the absence of cytotoxicity of (M-CA) _f and (M-CMD) _f towards these cells. This behavior corresponds to that observed with healthy mammalian cells. In contrast, in the presence of M-CA/M-CMD and a heating session, cell viability decreased by 30–40% and 60–85% after the application of AMF and LIU, respectively, by compared to the two controlled conditions in which cells are either only incubated with (M-CA) _f /(M-CMD) _f without heating or only exposed to AMF/LIU without exposure to NPs. These results suggest that LIU leads to more efficient cell killing than AMF, perhaps due to a temperature maintained at 46°C for a longer period with LIU than with AMF. Furthermore, it appears that although both types of formulated magnetosomes form an effective pair with LIU to destroy prostate tumor cells, M-CMD outperforms M-CA, a behavior we are currently trying to understand. Since internalization of NPs into tumor cells often correlates with efficient cell killing, we estimated the amount of (M-CA) _f and (M-CMD) _f , which is internalized into PC3-luc cells. after a treatment similar to that used to measure cell viability. After the treatments, we destroyed and dissolved the tumor cells, and measured their iron content by ICP-MS. The quantities of NP internalized in PC3-luc tumor cells, Q _i , are greater for (M-CMD) _f than for (M-CA) _f in all the conditions tested, that is to say with/without AMF/LIU treatments. Q _i is estimated to be 17-20 and 30 pg NP per cell after AMF/LIU treatment for (M-CA) _f and (M-CMD) _f , respectively. Compared to (M-CMD) _f , (M-CA) _f displays lower internalization into PC3-luc cells by a factor of 1.5-2. This behavior can be attributed to the more negative surface charge at physiological pH for (M-CMD) _f than for (M-CA) _f , which may result in a better affinity of (M-CMD) _f for the cationic sites of the plasma cell membrane and, subsequently, greater cellular internalization, perhaps by pinocytosis, for (M-CMD) _f than for (M-CA) _f . If we now consider the potential impact of NP internalization on cell viability, we can observe that increased NP internalization between M-CMD and M-CA either correlates with increased cell mortality after treatment with LIU, or not correlated with a change in cell mortality under AMF application. These behaviors could be explained by the fact that ultrasound acts in synergy with internalized NPs to destroy tumor cells, i.e. ultrasound could destroy/inactivate the cell membrane while NPs could act in a manner disruptive inside cells from their intracellular location. In contrast, while AMF is expected to produce local warming at the NP, it is not expected to act disruptively on the cell membrane. Thus, it is possible that in the presence of internalized NPs, ultrasound acts more disruptively than AMF, resulting in more efficient cell killing.

Claims (15)

Composition comprising at least one chain of at least two nanoparticles, in which each nanoparticle of the chain comprises an iron oxide mineral core surrounded by a coating,
in which the composition further comprises a cryoprotectant,
characterized in that the dissociation energy between the coating and the core is greater than the dissociation energy between the cryoprotectant and the core,
and in that the composition is in the form of a powder or a liquid suspension. Composition according to claim 1, in which the at least one chain is in a liquid suspension and the composition has at least one of the following properties:
- it is isotonic to animal plasma,
- the volume occupied by the water in the composition is greater than the volume occupied by the at least one chain of said composition, and
- the percentage by mass of water in the composition is greater than the percentage by mass of the at least one chain of said composition. Composition according to claim 1, in which the at least one chain is in powder form, and the composition has at least one of the following properties:
- the volume occupied by the water in the composition is smaller than the volume occupied by the at least one chain of said composition, and
-the percentage by mass of water in the composition is less than the percentage by mass of the at least one chain of said composition,
wherein the composition is preferably lyophilized, desiccated, dried or dehydrated. Composition according to one of claims 1 to 3, in which the cryoprotectant is chosen from the group consisting of: 1) Acetamide, 2) Acetate, 3) Albumin, 4) Amino acids, 5) Ammonium acetate, 6) Arginine , 7) Alcohols containing at least one or two hydroxyl groups, 8) Bridger, 9) Choline, magnesium chloride and sodium bromide, 10) Diethyl glycol, 11) Dimethylacetamide, 12) Dimethyl sulfoxide (DMSO), 13) Disaccharide, 14) Erythritol, 15) Ethanol, 16) Ethylene glycol, 17) Formamide, 18) Fructose, 19) Glucose, 20) Glycerol, 21) Glycerol 3-phosphate, 22) Glycol such as Diethyl glycol or Triethylene glycol, 23) Glycine, 24) Lactose, 25) L-tyrosine, 26) lysine hydrochloride, 27) Mannitol, 28) MDP (2-Methyl-2,4-pentanediol), 29) Phenylalanine, 30) Planic, 31) Polymers, 32) Polyethylene glycol tel as PEG4000, polyethylene glycol succinate, distearoylphosphatidyl ethanolamine-folate-modified polyethylene glycol, 33) Polyethyleneimine (PEI), 34) Polyvinylpyrrolidone (PVP), 35) Proline, 36) Propylene glycol, 37) Protein, 38) Pyridine (Pyridine -N-Oxide), 39) Ribose, 40) Sarcosine, 41) Serine, 42) Serum albumin, 43) Sodium bromide, 44) Sodium chloride, 45) Sodium dodecyl sulfonate, 46) Sodium glutamate, 47) Iodide sodium, 48) Sodium sulfate, 49) Sorbitol, 50) Starch (hydroxyethyl starch), 51) Sugar, 52) Sucrose, 53) Cell bank series, 54) Trehalose, 55) Triethylene glycol, 56) Trimethylamine, 57) Tween 80, 58) Tryptophan, 59) Valine, and 60) Xylose, and 61) a combination or derivative of any of these compounds. Composition according to one of claims 1 to 4, in which the mass percentage of cryoprotectant is between 0.5 and 50%. Composition according to one of claims 1 to 5, in which the core of the nanoparticle comprises a first center for producing or capturing free radicals C _{1 FRPC} , in which C _{1 FRPC} is preferentially chosen from the group consisting of:
- a metal other than iron, such as zinc or aluminum,
And
- a metal oxide other than iron oxide, such as zinc oxide or aluminum oxide. Composition according to claim 6, in which the coating comprises a second free radical production or capture center C _{2 FRPC} . Composition according to any one of claims 6 to 7, in which C _1FRPC and/or C _2FRPC is/are at least one antioxidant compound. Composition according to any one of claims 6 to 8, in which C _1FRPC and/or C _2FRPC is/are at least one oxidizing compound. Composition according to any one of claims 1 to 9, in which the core is synthesized by a living organism or cells producing nanoparticles, preferably a magnetotactic bacteria. Composition according to any one of claims 6 to 10, in which C _1FRPC and/or C _2FRPC is/are one or more photosensitizer(s), preferentially chosen from the group consisting of: 1) acridine, such such as acridine orange, acridine yellow, 2) ALA (5-aminolevulinic acid), 3) aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) aminolevulinic acid, delta acid -aminolevulinic, 5) Antihistamines, 6) Azulene, 7) Bavteriochlorine, 8) TOOKAD or TOOKAD Soluble, 9) WST-11, 10) LUZ11, 11) BC19, 12) BC21, 13) porphyrin such as Benzoporphyrin derivative monoacid ring A (BPD-MA), 14) Chlorine such as Chlorine e6, m-tetrahydroxyphenylchlorine 15) Foscan, 16) Verteporfin, 17) Monoacid A ring benzoporphyrin derivative, 18) Monoaspartyl chlorin(e6), 19) Talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrine, 23) Chlorine e6 porphrine, 24) Coal tar and derivatives, 25) Contraceptives, oral and estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview , 29) Colorants such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squaraines, 33) BODIPY dyes, 34) Phenalenones, 35) Benzophenoxazinium dyes, 36) Erythrosine, 37) Flavins, 38) Foscan , 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH), 45) Hypericin , 46) Hypocrellin, 47) ICG (Indocyanine Green), 48) Levulan, 49) MAL -methyl aminolevulinate), 50) Meta-tetra(hydroxyphenyl)chlorine (m-THPC), 51) Metvix, 52) Methylene blue, 53) Monoterpene, 54) Motexafin lutetium (Lu-Tex), 54) N-aspartyl chlorine e6 (NPe6), 55) Nanoparticle or nanomaterial, 56) Natural products or compounds, 57) Non-steroidal anti-inflammatory drugs, 58) Palladium bacteriopheophorbide (WST09), 59) Phthalocyanine-based dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phthalocyanine such as liposomal ZnPC, 65) Chloroaluminium sulfonated phthalocyanine (CASP), 66) Phthalocyanine from silicon (PC4), 67) RLP068, 68) Porfimer sodium, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-tetrakis(1-methylpyridinium-4-yl) tosylate porphyrin, 70) XF70 , 71) Protoporphyrin, 72) ALA-induced protoporphyrin IX, 73) Psoralens, 74) Quantum dots, 75) Quinones, 76) Riboflavin, 77) Rose Bengal, 78) silicon or silicon phthalocyanine (Pc4), 79) Sulfonamides, 80) Sulfonylureas, 81) Talaporfin or Talaporfin sodium, 82) Temoporfin, 82) Tetrahydropyrroles, 83) Tin ethyl etiopurpurin, 84) Titanium dioxide, 85) Toldudine blue O, 86) Transition metal compounds such as ruthenium (II), polypyridyl complexes, ruthenium, rhodium, cyclometalated bridged dimeric compounds Rh(II)-Rh(II), platinum (II), gold (III), 87) Verteporfin, 88) Compound with vulcan base such as aminovulinic acid, aminovulinic acid, 89) WST11, and 90) Xanthene. Composition according to any one of claims 6 to 11, in which C _1FRPC and/or C _2FRPC is/are sonosensitizer(s), preferably chosen from the group consisting of: 1) ABS-FA, 2 ) Acrylonitrile Butadiene Styrene, 3) Styrene, 4) Folic acid, 5) AIMP NP, aminoacyl tRNA synthetase complex-interacting multifunctional protein, 6) Au nanomaterial, 7) gold, 8) Au-MnO nanomaterial, 9) manganese oxide, 10) antineoplastic drugs, 11) NSAIDs, 12) nonsteroidal anti-inflammatory drugs, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) Au-doped TiO2, 17 ) Carbon-based nanomaterial, 18) Carbon nanotube, 19) Chlorine, 20) Ce6, 21) PTX, Paclitaxel, 22) drug or chemotherapeutic compound, 23) infrared dye or IR783, 24) Curcumin, 25) Cyanine or Cu -Cyanine, 26) DHMS, 27) dimethylsulfide, 28) Docetaxel, 29) drug or chemotherapeutic compound, 30) DOX/Mn-TPPS@RBCS, 31) doxorubicin, 32) manganese, 33) blood cell, 34) red blood cell, cell , 35) polymer, 36) elastomer, 37) erythosine or erythosine B, 38) FA or FA-OI or FA-OI NP or folic acid, 39) F3-PLGA@MB/Gd NPs, 40) poly(lactic acid- co-glycolic), 41) gadolinium, 42) Fe-TiO2 or titanium oxide, 43) Fe-VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49 ) hydrochloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysafflor yellow A, 55) Hypocrellin, Hypocrellin B, 56) IR780, 57) Levofloxacin, 58) LIP3 or Lithium phosphide , 59) Lithium, 60) Liposome or Liposomal nanomaterial, 61) Lomefoxacin,, 62) MG@P NPs, 63) MnP or Manganese peroxidase, 64) MnTTP-HSAs, 65) metal-porphyrin complex wrapped in HSA, 66) albumin , 67) MnWOx, 68) MnWOx-PEG, 69), PEG, 70) bimetallic oxide, 71) Mn (III)-HFs, 72) managen, hemoporphin, 73) Nano-beads, 74) Noble metal nanomaterial, 75 ) OI NP or oxygen indyocyanine, 76) Phthalocyanines, 77) PIO or Pioglitazone, 78) Polymeric nanomaterial, 79) Porphyrin, 80) TiO doped with Pt2, 81) R837, 82) Rose Bengal, 83) Sparfloxacin,, 84) TAPP or 5,10,15,20-tetrakis (4-aminophenyl) porphyrin, 85) TiO2 or titanium dioxide nanomaterial, 86) TCPP, isomer or Tris(1-chloro-2-propyl) phosphate 87) TPI or Thermoplastic Polyimide or thermoplastic polymer, 88) TPZ or Tirapazamine, 89) Transition metal oxide, 90) nanoparticle or Janus nanoparticle, and 91) Xanthones. Composition according to any one of claims 6 to 12, in which C1FRC and/or C2FRC is/are radiosensitizers, preferably chosen from the group consisting of: 1) AMG102, 2) AQ4N, 3) Apaziquone (E09), 4) Bromodeoxyuridine, 5) Carbogen, 6) Cetuximab, 7) Chemotherapy drug or compound, 8) Chlorpromazine, 9) C-reactive peptide, 10) Curcumin, 11) Diamide, 12) Diethylmaeate,, 13) Dihydroartemisinin, 14) Docetaxel , 15) ECI301, 16) Etanidazole, 17) Fludarabine, 18) 5-Fluorouracil, 19) Fluorodeoxyuridine, 20) Gadolynium, 21) Gemcitabine, 22) HER-3 ADC, 23) HSP, 24) Hydrogen peroxide, 25 ) Hydroxyurea, 26) Hyperbaric oxygen, 27) Hyperthermia, 28) Hypoxic cellular cytotoxic agent, 29) Irinotecan, 30) Radiosensitive lanthanide-based metal-phenolic network, 31) Lidocaine, 32) Lododeoxyuridine, 33) Metronidazole, 34) misonidazole, 35) etanidazole, 36) nimorazole, 37) N-Ethylmalemide, 38) malmeide, 39) ethylmalmeide, 40) Nanomaterials such as those consisting or composed at least partially or totally of gold, silver, bismuth, gadolinium, polysiloxane matrix and chelates of gadolinium, hafnium, tantalum, zinc, gadolinium, germanium, chromium, praseodymium, silicon, iron, platinum, cobalt, manganese, magnesium , iron, titanium, carbon nanotube, quantum dot, nanorad, triflate or metal oxide, 41) Nelfinavir, 42) Nicotinamide, 43) Nimotuzumab, 44) RNA, or miRNA, or miR-201 , or miR-205, or miR-144-5p, or miR-146a-5p, or miR-150, or miR-99a, or miR-139-5p, or miR-320a, 45) Active agent on membrane, 46 ) Mitomycin-C or Mitomycin, 47) Motexafin, 48) NBTXR3, 49) Oligonucleotide, 50) Paclitaxel, 51) Papaverine or papaverine hydrochloride, 52) Paraxonase-2, 53) Pocaine, 54) Porfiromycin (POR), 55) Protein, 56) Peptide, 57) Nucleosides or radiosensitizing compounds, 58) Resveratrol, 59) RRx-001, 60) SiRNa, 61) Sulfhydric group suppressants, 62) SYM004, 63) Texaphyrins, 64) TH-302, and 65 ) Tirapazamine. Process for manufacturing the composition according to any one of claims 1 to 13, which comprises at least one of the following steps:
Step 1 of amplification of magnetotactic bacteria in at least one medium, comprising:
the compounds necessary for the growth of magnetotactic bacteria and the production of magnetosomes, which are preferentially chosen from the group consisting of:
-a carbon source chosen from the group consisting of: at least one compound comprising at least one carbon atom, lactic acid, Na lactate, lactic acid, acetate, glycolate, glucose, pyruvate, succinate, carbon dioxide, glycerol and combinations thereof, at a concentration preferably between 1 nM and 2 Mol/L;
a source of iron chosen from the group consisting of: at least one compound comprising at least one iron atom, iron citrate, iron quinate, iron chloride, iron sulfate, FeCl3, and combinations thereof, a concentration preferably between 1 nM and 2.10-3 Mol/L;
a source of nitrogen chosen from the group consisting of: at least one compound comprising at least one nitrogen atom, a nitrate salt, gaseous nitrogen, ammonium, ammonia, a salt of ammonium, urea, an amino acid, ammonia gas, and combinations thereof, at a concentration preferably between 1 nM and 4 Mol/L;
-a source of oxygen chosen from the group consisting of: at least one compound comprising at least one oxygen atom, oxygen or air or compressed air, preferably in the form of a gas, the oxygen source being in certain cases bubbled or introduced into the growth medium, at a gas flow rate which is preferably between 5 ml of gas per minute and 50,000 ml of gas per minute;
a phosphate source consisting of at least one compound comprising at least one phosphate atom, at a concentration preferably between 1 nM and 2.10-1 Mol/L;
-a source of potassium consisting of at least one compound comprising at least one potassium atom, at a concentration preferably between 1 nM and 2.10-1 Mol/L;
-a source of sulfur or sulfate consisting of at least one compound comprising at least one sulfur or sulfate atom, at a concentration preferably between 1 nM and 4.10-1 Mol/L;
a source of manganese consisting of at least one compound comprising at least one manganese atom, at a concentration preferably between 1 nM and 4.10-1 Mol/L;
a source of vitamin chosen from the group consisting of: at least one compound comprising at least one vitamin, biotin, calcium, pantothenate, folic acid, inositol, nicotinic acid, p-aminobenzoic acid , pyridoxine HCl, riboflavin, thiamine, thiamine HCL and their derivatives and their combinations, at a concentration preferably between 1 nM and 10-4 Mol/L, and
-a source of calcium consisting of at least one compound comprising at least one calcium atom, at a concentration preferably between 1 nM and 10-1 Mol/L.
at least one compound necessary for doping the magnetosomes with C _1FRPC or a metal other than iron, preferably zinc or aluminum, for example a source of zinc, preferably zinc sulfate or zinc citrate or zinc chlorate or zinc quinate.
Step 2 of extracting magnetosomes from magnetotactic bacteria and purifying the magnetosomes extracted by heating to obtain magnetosome minerals comprising a mass percentage of organic matter originating from magnetotactic bacteria of less than 1%,
Step 3 of coating the magnetosome minerals with a coating material comprising the compound C _2FRPC by mixing the magnetosome minerals with the coating material, where the mixing is carried out under at least one of the following conditions:
under sonication,
under the application of radiation,
under temperature variation,
under pH changes,
under adjustment of the redox potential,
using a ratio between the quantity or mass of the magnetosome minerals and the quantity or mass of the coating material, preferably Compound D, which is adjusted or varied or greater than 1,
Step 4 consists of adding at least one cryoprotectant to the coated magnetosome minerals obtained at the end of step 3,
Step 5 of freeze-drying or dehydration or drying or desiccation of the composition obtained at the end of step 4,
Step 6 of resuspension of the lyophilized composition obtained in step 5, preferably in water. Process for storing the composition according to any one of claims 1 to 14, which comprises at least one of the following steps:
Step 1: Choose or prepare the composition in the form of a liquid suspension,
Step 2: Lyophilization, desiccation, dehydration of the composition, or elimination of water from the composition,
Step 3: Storage of the composition in powder form, preferably for more than 3 months,
And
Step 4: Suspending or resuspension of the composition, preferably in water.