NANOPARTICLES ARRANGED IN CHAINS IN AN ISOTONIC COMPOSITION AFTER RECONSTITUTION

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, wherein each nanoparticle in the chain comprises an iron oxide mineral core 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, and wherein the chain arrangement is maintained at a composition temperature below 0°C. Figure for abstract: No figure

Description

Title of the invention: NANOPARTICLES ARRANGED IN CHAINS IN AN ISOTONIC COMPOSITION AFTER RECONSTITUTION FIELD OF INVENTION

[0001] The field of the invention is that of nanoparticles, having a particular type of organization, such as a chain organization, the organization of which can be maintained for a long period, in particular to preserve the chains, by lyophilizing these nanoparticles in the presence of a cryoprotectant, keeping these lyophilized nanoparticles for a certain time, and resuspending the chains in a liquid such as water, preferably before their use or administration to humans.

[0002] CONTEXT OF THE INVENTION

[0003] Certain nanoparticles are known to organize themselves in a specific way. For example, magnetosomes synthesized by magnetotactic bacteria, which consist of iron oxide mineral cores coated with organic lipid bilayers, form chains (Applications of magnetotactic bacteria and the magnetosome for cancer treatment: A review emphasizing practical and mechanistic aspects, Edouard Alphandéry, Drug Discovery Today, Vol. 25, p. 1444, 2020). We were able to remove the lipid bilayer covering the mineral core of the magnetosomes and replace it with a synthetic coating (see reference above). However, the synthetic coating of the mineral core of the magnetosomes degrades in water over time. These nanoparticles cannot be maintained in a chain-like suspension for extended periods.We propose here a new product consisting of magnetosome chains mixed with a cryoprotectant, which are lyophilized for long-term storage as a powder. They are then resuspended in a liquid, where they reassemble into chains, before being used.

[0004] DESCRIPTION OF THE INVENTION

[0005] The invention relates to a composition comprising at least one chain of at least two nanoparticles, wherein each nanoparticle of the chain comprises a mineral core of iron oxide or a core of iron oxide or a metallic core or a core composed of a metallic oxide, surrounded by a coating, wherein the composition further comprises a cryoprotectant which is optionally mixed with water, wherein the dissociation energy between the coating and the core is preferably greater than the dissociation energy between the cryoprotectant and the nucleus, wherein the composition is preferably in the form of a powder or a liquid suspension, wherein the coating preferentially maintains the chain arrangement above 0°C, wherein the cryoprotectant preferentially maintains the chain arrangement below 0°C, wherein the chain arrangement is preferably maintained at a temperature below 0°C.

[0006] The invention also relates to a composition comprising at least two nanoparticles arranged in a geometric figure, wherein each nanoparticle of the geometric figure comprises an iron oxide mineral core surrounded by a coating, wherein the composition further comprises a cryoprotectant mixed preferably with water, wherein the dissociation energy between the coating and the core is preferably greater than the dissociation energy between the cryoprotectant and the core, wherein the cryoprotectant agent preferably serves to keep the geometric figure stable over a period of at least one or six months.

[0007] In some cases, the nanoparticle comprises a core and / or a coating, in which 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.

[0008] In other cases, the nanoparticle is amorphous and / or crystalline.

[0009] In some cases, an iron oxide nucleus is composed of at least one iron atom and at least one oxygen atom.

[0010] In some other cases, a nucleus composed of a metal oxide is composed of at least one metal atom and at least one oxygen atom.

[0011] In other cases, a metallic nucleus is composed of at least one metal atom.

[0012] In one embodiment of the invention, the composition comprises at least one nanoparticle with the cryoprotectant and optionally a liquid such as water.

[0013] In another embodiment of the invention, the composition comprises at least one nanoparticle without the cryoprotectant and optionally a liquid such as water.

[0014] In another embodiment of the invention, the composition comprises an excipient, an active ingredient, and / or a surfactant.

[0015] In another embodiment of the invention, the composition is a preparation or a suspension or a powder, preferably of or with at least one nano- particle, possibly with the cryoprotectant.

[0016] 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 preserves at least one property of the composition, preferably selected from: i) the chain arrangement or geometric figure of the at least two nanoparticles, ii) the at least one free radical capture or production center, 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.

[0017] In one embodiment, the free radical production or capture center is a compound that increases the amount 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, preferably designated as a thermosensitizer in this case, iv) cryogenic treatment or protection is applied to the center, preferably designated as a cryosensitizer in this case, v) a magnetic field is applied to the center, preferably designated as a magnetosensitizer in this case, vi) ultrasound is applied to the center, preferably designated as a sonosensitizer.

[0018] In one embodiment, the center for the production or capture of free radicals may be a center of physico-chemical perturbation, such as radiation, with amplification, where the amplification may correspond to an effect of the physico-chemical perturbation applied to the part of the body that is greater 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 whereas this effect would be absent or less pronounced in the absence of the center.

[0019] In one embodiment, the free radical production or capture center is a compound that amplifies or increases the physicochemical perturbation or radiation, or at least one parameter or property of the physicochemical perturbation or radiation, or at least one effect of radiation or a physicochemical perturbation. When the center is exposed to radiation, light, thermal variation, cryotreatment or cryoprotection, a magnetic field, or ultrasound, it may be designated respectively as a radiosensitizer, photosensitizer, thermosensitizer, cryosensitizer, magnetosensitizer, or sonosensitizer.

[0020] In another embodiment of the invention, at least one property of the The composition is maintained, preserved, or protected, preferably by the cryoprotectant, when: At least two nanoparticles remain arranged in a chain or geometric shape, at least one free radical production or capture center remains contained within the nanoparticle or composition or remains active, i.e., capable of preferentially producing or capturing free radicals under the application of 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 nanoparticle is maintained or decreased below or at a temperature of 10⁵, 10³, 10², 50, 20, 10, 5, 2, 1, 0, -5, -10, -50, -77, -100, -200, or -273. °C.

[0021] In some cases, Tl and T2 are two different temperatures.

[0022] In some cases, T2 is less than Tl, preferably by at least 10-5, 10-3, 10-1, 0, 1, 2, 5, 10 or 103 °C.

[0023] In some cases, Tl 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.

[0024] In some other cases, Tl and / or T2 is / are greater than -273, -200, -100, -77, -50, -10, -5, 0, 1, 2, 5, 10, 50, 100 or 103 °C.

[0025] In one embodiment of the invention, the cryoprotectant is not used to protect at least one nanoparticle or at least one chain against denaturation and / or destruction and / or loss of the primary, secondary, tertiary or quaternary structure of DNA, RNA, protein(s), lipid(s), enzyme(s) or at least one biological molecule.

[0026] In one embodiment of the invention, the cryoprotectant prevents a change, decrease, or increase in size of at least one nanoparticle or chain by more than 100, 50, 10, or 1%, where this percentage is preferably equal to S2-S1 / S1 or P2-P1 / P1, where SI, PI, S2, and P2 are preferably the sizes and properties of at least one nanoparticle or chain before (SI, PI) and after (S2, P2) the cooling of at least one nanoparticle or chain, preferably from more than 1, 50, or 100°C, preferably from an initial temperature, preferably above -273, -100, -50, 0, 5, or 10°C, preferably down to a final temperature, preferably below 100, 50, 0, or -50°C.

[0027] In some 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 core diameter of the at least one nanoparticle, vi) the coating thickness of the at least one nanoparticle, vii) the isotonicity or osmolality of the at least one nanoparticle or chain.

[0028] 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 to 10, 5, 2, 1, 0, -5, -10, -50, -77, -90, -270, or -273 °C, can be reached or obtained 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 remaining or existing. This effect is preferably due to the presence of the cryoprotectant.

[0029] In one embodiment of the invention, the composition is freeze-dried or desiccated, 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 being retained or existing. This effect is preferably due to the presence of the cryoprotectant.

[0030] In one embodiment of the invention, the composition is prepared or used by following at least one of the following steps:

[0031] First, the composition is mixed with water to add the cryoprotectant to at least one chain in liquid suspension.

[0032] Next, the composition is lyophilized to remove water from the composition and preserve the composition in powder form for storage.

[0033] Thirdly, the composition is resuspended in water, ready to be used and administered to humans.

[0034] In one embodiment, at least one of the steps mentioned above is repeated.

[0035] In one embodiment of the invention, the dissociation energy between a chemical group of a coating molecule and a chemical group of the nanoparticle nucleus is greater than 10 Kcal / mol, 100 KJ / mol or 1 eV / bond.

[0036] In one embodiment of the invention, the dissociation energy between: i) a chemical group of a cryoprotectant molecule and a chemical group of the nanoparticle core or ii) a chemical group of a cryoprotectant molecule and a chemical group of the nanoparticle coating, is less than 10 Kcal / mol, 100 KJ / mol, or 1 eV / bond.

[0037] 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 chemical affinity between the cryoprotectant and the nanoparticle nucleus.

[0038] In one embodiment of the invention, the coating is more strongly bound to the core of the nanoparticle or interacts more strongly with it than the cryoprotectant.

[0039] In one embodiment of the invention, the coating cannot be separated or isolated from the nanoparticle core or is inseparable or non-isolable from the nanoparticle core, preferably by using a magnet or magnetic separation or centrifugation.

[0040] In another embodiment of the invention, the cryoprotectant can 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.

[0041] In some cases, this can be demonstrated by the fact that: i) the coating and the core of the nanoparticle cannot be separated by a low-intensity magnet, i.e. preferably of less than 1 T or 1 mT, or of low magnetic field gradient, i.e. preferably of a magnetic field gradient of less than 1 T or mT per cm preferentially of the body part, whereas 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 whereas the cryoprotectant can be preferentially separated from the nanoparticle by being mixed in water with the nanoparticles in suspension followed by centrifugation.

[0042] In one embodiment, the coating comprises at least one compound or chemical function capable of establishing interactions, weak interactions, weak bonds, or covalent bonds with the core of the nanoparticle, or at least one chemical function, atom, or ion from 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 keep the coating of the nanoparticle attached, bound, or associated with or to the core of the nanoparticle, preferably referred to as association, attachment, or bonding interactions.

[0043] In one embodiment of the invention, the nanoparticle consists of a core, a surrounding coating and association or bonding bonds or interactions.

[0044] In one embodiment of the invention, the nanoparticle consists of a core and a surrounding coating, which is preferably associated, bound, or attached to the core, preferably in a manner or by sufficiently strong bonds or interactions strong so that it does not detach or dissociate from the nucleus or so that it remains attached or bound to the nucleus.

[0045] In some cases, the chain arrangement or the geometric figure of the nanoparticles is maintained or exists or is formed in the presence of the coating.

[0046] In some cases, in the absence of coating, the chain arrangement or geometric figures do not exist or do not form or do not exist.

[0047] In one embodiment of the invention, the composition comprises a cryoprotectant, which is associated or bound or linked to the nanoparticle, the core, the coating, preferably in a manner or by bonds or interactions sufficiently weak that it detaches or dissociates from the nanoparticle, the core or the coating, preferably when the cryoprotectant is carried away or removed from the nanoparticles by washing with water, by centrifugation or by using a magnet.

[0048] In some cases, the cryoprotectant can be dissociated or detached or disassembled or unbound from the nanoparticle, the core or the coating, preferably in such a way that the chain arrangement or geometry of the nanoparticles is maintained.

[0049] In certain cases, the cryoprotectant may be associated, attached, assembled, or bonded to or with the nanoparticle, core, or coating, preferably in such a way that the chain arrangement or geometry of the nanoparticles is maintained, preferably when the composition is cooled, preferably below or at 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 water is partially or totally removed from the composition, or when the mass percentage of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3, or 10-5 %, or when the The composition is in powder form.

[0050] 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) deposition of a droplet of the composition on a substrate, preferably a carbon grid, iv) waiting for the water to evaporate, and v) observation of the arrangement of the nanoparticles under the electron microscope.

[0051] In some cases, preferably, in the absence of coating, the chains or geometric figures do not form or do not exist, while in the presence of coating, the chains or geometric figures preferably form or exist.

[0052] In certain cases, when the temperature of at least one nanoparticle is less than or equal to 100, 10, 5, 2, 1, 0, -10, -50, -77, -270 or -273 °C, preferably for more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the nanoparticle is lyophilized or dried or when water is removed from the composition or of the at least one nanoparticle partially or totally or when the mass percentage 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 geometric figure does not preferentially exist or does not form or is destroyed in the absence of the cryoprotectant, and / or the chain arrangement or geometric figure preferentially exists or forms or is maintained or is not destroyed in the presence of the cryoprotectant.

[0053] 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 bound or associated or bound to the core of the nanoparticle, preferably in the presence of the cryoprotectant, surrounding or 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 mass percentage of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5%.

[0054] 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 to the core of the nanoparticle, preferably in the absence of the cryoprotectant, surrounding or 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 mass percentage of water in the composition is less than 100, 50, 10, 5, 2, 1, 0, 10-3 or 10-5%.

[0055] 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, hydroxyls OH-, oxides O2-, crystalline defects of the core, which may be located in or on the surface of the nanoparticle core.

[0056] In one embodiment, the coating comprises at least one compound, atom, ion or chemical function such as an acid function, carboxylic acid, phosphoric acid or sulfonic acid, wherein the compound, atom or 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, an ion of the nucleus such as Fe2+, Fe3+, Hydroxyl OH-, oxide O2- or a crystal defect of the nucleus.

[0057] In one embodiment of the invention, the interactions or bonds exist or are maintained when the composition is cooled, preferably below or at 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 totally or when the mass percentage 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 some cases, these interactions or bonds maintain the nanoparticles organized in a chain or geometric shape; that is, preferentially, in the absence of these interactions or bonds, the nanoparticles are not organized in a chain or geometric shape.

[0058] In some cases, a geometric shape is an assembly or aggregate of nanoparticles forming a geometric shape.

[0059] In some 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, a rational triangle, a right triangle, a Kepler triangle, a Scalene triangle, a quadrilateral, a cyclic quadrilateral, a kite, a parallelogram, a rhombus, a rhomboid, a rectangle, a square, a tangential quadrilateral, a trapezoid, an isosceles trapezoid, a pentagon, a hexagon, a Lemoine hexagon, a heptagon, an octagon, a nonagon, Decagon, Hendecagon, Dodecagon, Tridecagon, Tetradecagon, Pentadecagon, Hexadecagon, Heptadecagon, Octadecagon, EnneadecagonIcosagon, Swastika, Star polygon, Pentagram - star polygon, Hexagram, Star of David, Heptagram, Octagram, Star of Lakshmi, Decagram - star polygon, Annulus, Arbelos, Circle, Archimedean twin circles, Bankoff circle, Circumference, Disk, Incircitation and excircle of a triangle, Nine-pointed 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. ,

[0060] In one embodiment of the invention, the nanoparticle, suspension, composition or assembly of nanoparticles is stable, preferably for a period of time, preferably its stability time, 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.

[0061] In some cases, nanoparticles, nanoparticle suspensions, nanoparticle compositions or assemblies may be stable at a nanoparticle concentration particles greater than 1, 5, 10, 50, 100, 200, 500 or 1000 mg of nanoparticles per mL of solvent, water, matrix or body part surrounding or comprising or incorporating the nanoparticles or composition.

[0062] In one embodiment of the invention, the body part is the body part of an individual, an animal, or a human.

[0063] In one embodiment of the invention, the body part comprises more than or at least 1, 2, 5, 10 or 100 similar or different organism(s), apparatus(s), organ(s), tissue(s), cell(s) or bio-molecule(s).

[0064] 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.

[0065] In some other cases, the body part may be or belong to an organ, the musculoskeletal, muscular, digestive, respiratory, urinary, female reproductive, male reproductive, circulatory, cardiovascular, endocrine, circulatory, lymphatic, nervous (peripheral or not), ventricular, enteric nervous, sensory, or integumentary system, a reproductive organ (internal or external), a sensory organ, or endocrine glands. The organ or body part can 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, testicles, 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, intestinal-associated lymphoid tissue, tonsils, brain, cerebrum, cerebral hemispheres, diencephalon, brainstem, midbrain, pons, medulla oblongata, cerebellum, spinal cord, choroid plexus, nerves, cranial nerves, spinal nerves, ganglia, eye, cornea iris, ciliary body, lens, retina, ear, external ear, earlobe, eardrum, middle ear, ossicles, inner ear, cochlea, vestibule of the ear, semicircular canals, olfactory epithelium, tongue, taste buds,mammary glands or skin.

[0066] In some cases, the body part or organ may belong to the bloodstream or circulatory system.

[0067] In some cases, the body part may be or include at least one tumor, cancer, virus, bacterium or pathological cell.

[0068] In one embodiment of the invention, the body part is or comprises water, an excipient, a solution, a suspension, at least one chemical element, organic matter or a gel, which may be synthetic or produced by a living organism.

[0069] 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 an organism, preferably a living or inactivated or dead organism, comprising at least one prokaryotic or eukaryotic cell.

[0070] In one embodiment of the invention, the body part 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 from or extracted from that region.

[0071] In one embodiment of the invention, the body part comprises a pathological site, a healthy site, and / or a region of nanoparticles or composition.

[0072] In one embodiment of the invention, the body part is or includes a pathological site or pathological cells.

[0073] 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 other pathological material. Pathological cells may be cells that: i) are not arranged or do not function as they normally do in a healthy individual, ii) divide more rapidly than healthy cells, iii) are healthy cells that have undergone a 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 may eventually penetrate, colonize, or replicate within these cells.In some cases, pathological cells can be likened to viruses or other organisms or entities that colonize, target, destroy, utilize, or interact with cells, preferably to enable their own reproduction, multiplication, survival, or death. In some cases, a pathological site may include healthy cells, the number, activity, or proliferation of which is preferably lower than that of the pathological cells.

[0074] In one embodiment of the invention, the body part is or comprises a healthy site or healthy cells. In some cases, the healthy site may be defined as a site or region that comprises one or more healthy cells, a healthy cell being defined as a cell that belongs to a healthy individual or to the body part of a healthy individual.

[0075] In some cases, the healthy site may surround the pathological site when it is preferentially located at a distance of less than 1 or 10-9 m from the pathological site.

[0076] In some cases, the composition or part of the body may be exposed to radiation.

[0077] Preferably, the radiation is chosen from the group consisting of: i) a magnetic, electric or electromagnetic field or wave, a wave or particle 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) a microwave, viii) radio frequencies, and ix) sound, ultrasound, infrasound or an acoustic wave.

[0078] In one embodiment of the invention, the radiation exhibits 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 cm², W per cm², or W per cm³, preferably of the part of the body or composition, or preferably per gram or milligram, preferably of the part of the body or composition, (iil) 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 of 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 cm², 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.

[0079] In another embodiment of the invention, the radiation exhibits 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 per cm², W per cm², or W per cm³, preferably per part of the body or composition, or preferably per gram or milligram, preferably per part of the body or composition, iil) 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 cm², 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 of the body part 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, preferably per cm², per cm², or per cm³, preferably of the body part or of the composition, preferably per gram or milligram, preferably of the composition or of the body part, iii) it has a frequency less 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 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.

[0080] In one embodiment of the invention, the nanoparticle or composition is administered to or into the body part.

[0081] 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.

[0082] 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.

[0083] In another embodiment of the invention, the nanoparticle or composition is administered to or into the following part of the body by at least one of the following routes of administration: local, enteral, gastrointestinal, parenteral, topical, oral, inhalation, intramuscular, subcutaneous, intratumoral, into an organ, into a vein, into arteries, into the blood or into a tissue.

[0084] In some cases, nanoparticles, nanoparticle suspensions, nanoparticle compositions or nanoparticle assemblies can be stable when at least one nanoparticle is not degraded or does not partially or totally lose its coating or can be administered to a part of the body or retains or maintains its chain arrangement or geometric shape.

[0085] In other cases, the nanoparticles, the nanoparticle suspension, the nanoparticle composition or assembly can be stable when the optical density of the nanoparticles, the nanoparticle suspension, the nanoparticle composition or assembly, preferably mixed in water, preferably measured at 480 nm or at 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 the 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 optical density measurement or absorption measurement of this suspension or composition.This percentage can be equal to (ODB - ODA ) / ODB or ODA / ODB , where ODB is the optical density of the nanoparticles, nanoparticle suspension, composition, or nanoparticle assembly measured before homogenization or mixing or optical density measurement or absorption measurement of the nanoparticle, suspension, composition, or nanoparticle assembly and ODA is the optical density of the nanoparticles, nanoparticle suspension, composition, or nanoparticle assembly measured after homogenization or mixing or optical density measurement or absorption measurement of the nanoparticles, nanoparticle suspension, composition, or nanoparticle assembly.

[0086] In some cases, the composition may be stable or considered stable when it is measured as stable at a certain first time t0 and at a certain second time tl, where tl follows t0 or is separated from t0 by a time interval A T of at least 1 second, 1 minute, 1 hour, 1 day, 1 month, 3 months, 6 months, 1, 2, 5 or 10 years. During A T, the composition is preferably stored.

[0087] 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.

[0088] In one embodiment of the invention, the cryoprotectant serves to maintain the chain or geometric figure stable for a period of time, preferably AT, preferably at least one or six months. In this case, the chain or geometric figure can preferably be observed or measured, preferably by electron microscopy, at a first time t0 and at a second time tl, where tl is preferably separated from t0 by AT.

[0089] 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.

[0090] In some 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.

[0091] 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 composition temperature preferably greater than or equal to -273, -200, -100, -50, -20, -10, -5, 0, 1, 2, 5, 10, 50, 100, 500, or 103 °C.

[0092] In some 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 greater than 10-10,10-5, 10-3, 10-1,0, 1, 2, 5, 10, 102,103 or 105 minutes or seconds.

[0093] In some 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.

[0094] The invention relates to the composition according to the invention, wherein at least one chain is in liquid suspension and the composition preferably has at least one of the following properties: i) it is isotonic to animal or human plasma or blood, (ii) the volume occupied by water in the composition is greater than the volume occupied by at least one chain in the composition, and iii) the mass percentage of water in the composition is greater than the mass percentage of at least one chain in the composition.

[0095] In one embodiment of the invention, 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.

[0096] In some cases, the composition is isotonic, that is to say, it preferentially exhibits: the same osmotic pressure as that of a body part such as a cell, a bodily fluid, plasma, blood, preferably from 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.

[0097] 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.

[0098] 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.

[0099] 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 body part.

[0100] In some cases, the composition may be hypertonic, that is to say, preferably in this case, the composition causes the shrinkage of at least one cell or part of the body.

[0101] In other cases, the composition may be hypotonic, that is to say, preferably in this case, the composition causes at least one cell or part of the body to swell.

[0102] In other cases, the composition may be isotonic, that is to say, preferably in this case, the composition produces no change in the volume of cells or parts of the body or a lesser change in the volume of cells or parts of the body than for a hypertonic or hypotonic composition.

[0103] In one embodiment of the invention, the solutes or solute compounds in the composition, preferably the nanoparticle, the nanoparticle chain and / or the cryoprotectant, have a concentration, preferably designated by Csolute, outside a cell or outside a body part, preferably designated by Csolute, inside a cell or inside a body part, 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 %.

[0104] 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%.

[0105] 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 body part, is the same as the osmolarity or osmotic pressure of an intracellular medium or body part or of a composition contained within at least one cell or body part.

[0106] In some cases, the body part may refer to a fluid or liquid medium contained within a body part, such as the intracellular medium or plasma or blood.

[0107] In some cases, the composition is included in the body part, preferably after or with administration to the body part.

[0108] In some other cases, the composition is contained outside the body part, preferably before or without administration to the body part.

[0109] In some cases, the composition may be hypo-osmotic.

[0110] In some other cases, the composition may be hyperosmotic.

[0111] 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.

[0112] 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.

[0113] 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% of the osmolality of human plasma, preferably between 275 and 299 milli-osmoles per kilogram.

[0114] In another embodiment of the invention, hydrostatic pressure or osmotic pressure or arterial pressure or systolic pressure or diastolic pressure or pressure of the composition or in the presence of the composition or following administration of the composition or applied by the composition to the body part or the force pushing the composition or at least one compound or solute of the composition preferably per unit area of ​​the body part, preferably in some cases from outside the body part or from at least one cell of the body part towards the inside of the body part or towards the inside of at least one cell of the body part, preferably in other cases from inside the body part or from the inside of at least one cell of the body part towards the outside of the body part or towards the outside of at least one cell of the body part, is designated by .

[0115] In some cases,^ 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.

[0116] In certain other cases,^ 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.

[0117] 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 includes the cryoprotectant. In this case, the volume of liquid is preferably maintained 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 motion of the at least one chain in the composition.

[0118] In some cases, the volume occupied by the water or liquid and preferably the cryoprotectant in the composition is greater than the volume occupied by at least one chain in the composition, by a factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

[0119] In some other cases, the volume occupied by the water or liquid and preferably the cryoprotectant in the composition is less than the volume occupied by at least one chain in the composition, by a factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in powder form.

[0120] In some cases, a may be equal to Vliq / Vchain, where Vliq and Vchain are the volumes occupied by the liquid preferentially including the cryoprotectant and at less one chain in the composition, respectively, where both volumes can preferably be measured by separating the at least one chain from the liquid, using for example magnetic separation or using a magnet to attract the chains and separate them from the liquid.

[0121] 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 includes the cryoprotectant. In this case, the mass of liquid is preferably maintained 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 motion of the at least one chain in the composition.

[0122] In some cases, 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, by a factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

[0123] In some other cases, the mass of water or liquid and preferably of cryoprotectant in the composition is less than the mass of at least one chain in the composition, by a factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in powder form.

[0124] In some cases, a may be equal to mliq / mchain, where mliq and mchain are the masses of the liquid comprising preferably the cryoprotectant and at least one chain in the composition, respectively, wherein the two masses can be preferably measured by separating the at least one chain from the liquid, using for example magnetic separation or using a magnet to attract the chains and separate them from the liquid, where mchain and / or mliq can be preferably measured by or after evaporation of the liquid from the composition or freeze-drying or drying of the composition.

[0125] In one embodiment of the invention, the mass percentage of water or liquid, preferably comprising the cryoprotectant, preferably not comprising at least one chain, in the composition, is greater than the mass percentage of at least one chain in the composition.

[0126] In some cases, the mass percentage of water or liquid, preferably comprising the cryoprotectant and preferably not comprising at least one chain, in the composition is greater than the mass of at least one chain in the composition by a factor of at least 1.1, 2, 5, 10 or 103. In this case, the composition is preferably in liquid form.

[0127] In one embodiment of the invention, the mass percentage of water or liquid, preferably comprising the cryoprotectant, preferably not comprising at least one chain, in the composition, is less than the mass percentage of at least one chain in the composition by a factor of at least 1, 1, 2, 5, 10 or 103. In this case, the composition is preferably in powder form.

[0128] In some cases, a may be equal to Pliq / Pchain, where Pliq and Pchain are the for mass percentages of the liquid comprising preferably the cryoprotectant and at least one chain in the composition, respectively, where both mass percentages can preferably be measured by separating the at least one chain from the liquid, using for example magnetic separation or using a magnet to attract the chains and separate them from the liquid, where Pchain and / or Pliq can preferably be measured by or after evaporation of the liquid from the composition or lyophilization or desiccation of the composition.

[0129] The invention also relates to the composition according to the invention, wherein at least one chain is in powder form, and the composition preferably has 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 at least one chain in the composition, and ii) the mass percentage of water or liquid and preferably cryoprotectant in the composition is less than the mass percentage of at least one chain in the composition, in which the composition is preferably freeze-dried, desiccated, dried or dehydrated.

[0130] In some cases, the composition is freeze-dried, desiccated, dried or dehydrated when the water or liquid is removed, preferably partially or totally, from the composition.

[0131] The invention also relates to the composition according to the composition, in which the composition is freeze-dried or desiccated or dried or dehydrated or treated by freeze-drying or desiccation or removal of water or removal of liquid or dehydration.

[0132] In one embodiment of the invention, the composition is dehydrated or the water or liquid is removed or absent from the composition. In this case, the mass percentage of water or liquid in the composition is preferably greater before the dehydration of the composition or before the removal of the water or liquid from the composition than after the dehydration of the composition or after the removal of the water or liquid from the composition. In this case, the volume of water or liquid The area 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 the water or liquid is removed from the composition. In this case, the water or liquid is removed from the composition without removing at least one chain and preferably also the cryoprotectant from the composition.

[0133] In one embodiment of the invention, the mass percentage of water in the composition is greater than 10-3, 10-1, 0, 1.5, 10, 25, 50, 80 or 99%, preferably before the water is or has been removed from the composition.

[0134] In another embodiment of the invention, the mass percentage of water in the composition is less than or equal to 100, 99, 80, 50, 25, 10, 5, 2, 1 or 0%, preferably after the water has been or is removed from the composition.

[0135] In another embodiment of the invention, the composition is lyophilized 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 at least one chain and / or the cryoprotectant from the composition.

[0136] 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 at least one chain and / or the cryoprotectant from the composition.

[0137] In one embodiment, the composition is contained in a tablet or is presented in the form of a tablet or powder, preferably after removal of water from the composition.

[0138] In one embodiment, the composition is stored or kept or unused, preferably for more than 0, 1, 10, 102, 103, 105, or 1010 hour(s), day(s), month(s), 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 nanoparticle core, partially or totally.

[0139] In one embodiment of the invention, the moisture 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.

[0140] In another embodiment of the invention, the moisture 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 as water by weight of the composition or as water by weight of 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.

[0141] The invention also relates to the composition according to the invention, wherein 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 preferably removed from the composition before the latter is suspended or resuspended in a liquid or water.

[0142] The invention also relates to the composition according to the invention, wherein the coating is selected from the group of compounds consisting of: 1) citric acid, 2) oleic acid, 3) polymethacrylic acid, 4) poly(ethylene oxide)-β-poly(methacrylic acid), 5) polyacrylic acid (PAA), 6) polylactic acid, 7) poly(ethylene oxide)-β-poly(glutamic acid), 8) phosphonic acid, 9) albumin, 10) alendronate, 11) alginate, 12) Au, A12 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 Dioleoylphosphati-dylcholine or phospholipid, 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) Mn3O4, 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) Poly(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-DL lactide, 63) Polyal-kylcyanoacrylate,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) Polymethyl methacrylate, 70) Polymersome, 71) Polystyrene, 72) PVA or polyvinyl alcohol, 73) PVP or polyvinylpyrrolidone, 74) Silica, preferably amorphous or mesoporous, 75) Silane, 76) SiO2, 77) Sodium oleate, 78) Starch, 79) Styrene, preferably styrene-divinylbenzene, 80) TaOx, 81) ZrO2, 82) at least one metal or semi-metal, 83) at least one metal oxide or 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 one 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.

[0143] The invention also relates to the composition according to the invention, wherein the coating is selected from the group of compounds consisting of: i) polysaccharides such as agarose, alginate, carregeenan, chitosan, dextran, haparin, gum arabic, pullulan, and starch; ii) acids such as citric acid, oleic acid, polymethacrylic acid, poly(ethylene oxide)-β-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 acid function; iii) polymers such as dextran, poly(ethylene oxide), Poly(vinyl alcohol), Poly(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, Al2O3, ZrO2, ferrites, MnO, Mn3O4, Au, Bentonite, carbon such as inactivated, activated, graphited 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 Alcohol chemical function, xvii) compounds comprising the Amide chemical function, xviii) compounds comprising the Aldehyde chemical function. ,

[0144] The invention also relates to the composition according to the invention, in which the coating has at least one chemical group selected 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.

[0145] The invention also relates to the composition according to the invention, in which the coating or coating material comprises a chemical function, preferably preferably chosen from the group consisting of i) OH-, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acidic derivative of at least one of these functions, which has an interaction or forms a chemical bond with an atom or 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 charge difference between the coating and the surface or nucleus 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.

[0146] In one embodiment of the invention, the cryoprotectant interacts or forms hydrogen or van der Wall or London bonds or interactions with the core or coating of the nanoparticle, preferably when the water molecules are displaced or cooled.

[0147] In one embodiment of the invention, as the cryoprotectant replaces 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.

[0148] In one embodiment, the composition including the cryoprotectant does not include ice, while the composition not including the cryoprotectant includes ice, or the composition including the cryoprotectant includes more ice than the composition not including the cryoprotectant, preferably when the composition is cooled and / or lyophilized,

[0149] 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.

[0150] In one embodiment of the invention, the cryoprotectant is a non-permeable cryoprotectant.

[0151] In some cases, the cryoprotectant may be chosen from: a sugar, trehalose, sucrose, starch, hydroxyethyl starch, polyvinylpyrrolidone and / or polyethylene oxide.

[0152] In some cases, the cryoprotectant may be a non-permeable cryoprotectant, that is to say, it does not preferentially penetrate cells or coatings and therefore remains preferentially extracellular or outside the coating when the composition is cooled.

[0153] In some cases, the composition is cooled during or for the cryopreservation of the composition.

[0154] In some cases, a non-permeable cryoprotectant is used to protect cells during the cooling of the composition.

[0155] Preferably, when cooling, slow cooling, or slow freezing is applied to the composition, or when the composition is cooled, preferably slowly, sufficient time is allowed for the water or liquid contained within the nanoparticle, preferably within the nanoparticle coating, to leave the coating, preferably under the effect of osmotic pressure, which preferably results in a reduction in the coating's volume. In the absence of a cryoprotectant, such behavior can destroy or damage the coating and / or lead to the destruction of the nanoparticle's chain arrangement.

[0156] In one embodiment, the cryoprotectant is permeable. Preferably, a permeable cryoprotectant can penetrate a cell or a coating and preferably preserve the cell or coating from osmotic pressure or damage or destruction.

[0157] In some cases, the cryoprotectant may be dimethyl sulfoxide (DMSO), glycerol, ethylene glycol or propylene glycol.

[0158] In some cases, the cryoprotectant can be used to induce vitrification of the intracellular environment or of the coating or of the internal or external part of the coating or of the nanoparticle or of the nucleus, preferably before the formation of ice crystals, preferably in the composition, preferably by preventing excessive volume loss of the cell or of the coating or of the nanoparticle or of the nucleus.

[0159] In one embodiment of the invention, the cryoprotectant is chosen from the group consisting of: i) a natural product, ii) a soy flour-based product, iii) a carbohydrate, iv) a lipid, v) a saccharide, vi) a zwitterionic molecule, vii) 1-camitin, and viii) antifreeze compounds such as antifreeze proteins.

[0160] The invention also relates to the composition according to the invention, wherein the coating or at least one of its chemical functions or atoms forms a first type of interaction or chemical bond with an atom or chemical function of the nucleus, wherein the cryoprotectant or at least one of its chemical functions or atoms forms a second type of interaction or chemical bond with the coating or at least one of its chemical functions or atoms, wherein the first type of interaction or chemical bond is preferably different from the second type of interaction or chemical bond, wherein the first type of interaction or chemical bond is preferably chosen from the group consisting of: i) an electrostatic interaction or bond, that is to say, preferably due to the charge difference between the coating and the surface or the nucleus, 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, in which the second type of interaction or chemical bond is preferably a hydrogen, London, or Van Der Walls interaction or bond.

[0161] In some cases, the coating or coating material may include a chemical function, preferably chosen from the group consisting of i) OH-, ii) NH2 , iii) COOH, iv) Thiol, v) Phosphate, and a basic or acidic derivative of at least one of these functions.

[0162] In some other cases, the nucleus may include a chemical function, preferably located on the surface of the nanoparticle nucleus, preferably a hydroxyl group (Fe-OH).

[0163] The invention also relates to the composition according to the composition, in which the cryoprotectant is selected 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 bromide, 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 acid, 31) Polymers, 32) Polyethylene glycol such as PEG4000, polyethylene glycol succinate, distearoylphos-phatidyl 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 dodecylsulfonate, 46) Sodium glutamate, 47) Sodium iodide, 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.

[0164] In one embodiment of the invention, the composition is desiccated, freeze-dried, dehydrated, or dried. In this case, the mass percentage 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 amount 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.

[0165] In some other cases, the composition may be undried, undehydrated, or undehydrated and dried. In this case, the mass percentage 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.

[0166] The invention also relates to the composition according to the invention, which is freeze-dried or dried or dehydrated, in which the mass percentage of cryoprotectant is preferably greater than the mass percentage of coating in the freeze-dried or dried or dehydrated composition, in which the freeze-dried or dried or dehydrated composition is preferably more stable than the non-freeze-dried or non-dried or non-dehydrated composition.

[0167] The invention also relates to the composition, where the mass percentage of the cryoprotectant is between 0.5 and 50%.

[0168] 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 the substances included in the composition.

[0169] In some cases, the mass percentage of the cryoprotectant, nanoparticle coating, nanoparticle core, at least one chain, 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%.

[0170] In some other cases, the mass percentage of the cryoprotectant, nanoparticle coating, nanoparticle core, at least one chain, liquid or water, in the composition is less than 100, 75, 50, 25, 10, 5, 2, 1, 0, 10-3 or 10-5%.

[0171] 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 water, in the composition.

[0172] 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.

[0173] The invention also relates to the composition according to the invention, in which the core of the nanoparticle comprises a first center for the production or capture of free radicals CiFrPC, where Cifrpc is preferentially chosen from the group consisting of: (i) a metal other than iron, such as zinc or aluminum, and (ii) a metallic oxide other than iron oxide, such as zinc oxide or oxide aluminum.

[0174] In some cases, CiFRPC may comprise at least one atom that is preferentially bonded to at least one oxygen or iron atom of the nucleus through at least one metallic bond.

[0175] The invention also relates to the composition according to the invention, in which the coating comprises a second center for the production or capture of C2 free radicals FRPC •

[0176] The invention also relates to the composition according to the invention, in which C2FRPC is linked to at least one atom of the coating, preferably by at least one covalent bond.

[0177] In one embodiment of the invention, the center for the production or capture of free radicals is at least one atom, molecule, chemical function, ionized or non-ionized, charged or uncharged, which produces or captures at least one free radical.

[0178] In one embodiment of the invention, a free radical is at least one atom, molecule, ion or chemical function that has: i) at least one unpaired valence electron, ii) an ability to dimerize, iii) a short lifetime, and / or iv) two unpaired electrons.

[0179] In one embodiment of the invention, a free radical is at least one atom, molecule, ion or chemical function that is: i) a chemically reactive species, ii) a hydroxyl radical (HO-), iii) a triplet oxygen, iv) a triplet carbene, and / or v) (❖CH2).

[0180] 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 change in pH.

[0181] In one embodiment of the invention, the free radical production center is at least one atom, molecule, or chemical function, ionized or non-ionized, charged or uncharged, which produces at least one free radical, preferably such 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 including the free radical production center as for the composition not including the free radical production center.

[0182] In one embodiment of the invention, the free radical capture center is at least one atom, molecule, or chemical function, ionized or non-ionized, charged or uncharged, that captures at least one free radical, preferably such that the amount or concentration of free radicals present when the composition includes the free radical capture center is less than the amount or concentration of free radicals present when the composition does not include the free radical capture center, where the comparison is preferably carried out using the same or similar excitation conditions for the composition including the free radical capture center as for the composition not including the free radical capture center.

[0183] The invention also relates to the composition according to the invention, wherein CiFRPC and / or C2FRPC is / are at least one antioxidant compound.

[0184] The invention also relates to the composition according to the invention, in which CiFRPC and / or C2Frpc is / are at least one oxidizing compound.

[0185] In one embodiment of the invention, the core of the nanoparticle in the composition according to the invention has at least one property selected from the group consisting of: a) it is ferrimagnetic, b) it is composed of maghemite or magnetite or of a composition intermediate between maghemite and magnetite, c) it has a size between 0.1 and 100 nm, and d) it comprises at least one crystallographic plane.

[0186] In another embodiment of the invention, the coating of the nanoparticle in the composition according to the invention has at least one property selected from the group consisting of: a) it has a thickness of less than 10 pm, 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 of less than 10 nm or the diameter of the nucleus.

[0187] In another embodiment of the invention, the cryoprotectant in the composition according to the invention is included in a matrix or volume encompassing the core and coating of the nanoparticle.

[0188] The invention also relates to the composition according to the invention, in which the nucleus or core of the nanoparticle is synthesized by a living organism or nanoparticle-producing cells, preferably a magnetotactic bacterium.

[0189] In one embodiment of the invention, the nanoparticles are synthesized biologically or by a living organism, referred to as the synthesizing living organism, which preferably consists or comprises at least 1, 2, 5, 10, 10³, 10⁶, or 10⁹ eukaryotic cell(s), prokaryotic cell(s), or a portion thereof. In some cases, a portion 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, or endoplasmic reticulum. rough, 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 can 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, 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.

[0190] In some cases, the living synthesizing organism may be magnetotactic bacteria, other types of bacteria than magnetotactic bacteria or enzymes from certain bacteria, preferably synthesizing nanoparticles extracellularly, such as Mycobacterium paratuberculosis, Shewanella oneidensi, Geothrix fermentans, ants, fungi or various plants.

[0191] 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, organelle or other biological material, such as a protein, lipid, enzyme, DNA or RNA, which is preferably produced by or derived from a eukaryotic or prokaryotic cell.

[0192] 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 such a cell.

[0193] In another embodiment of the invention, the nanoparticles are synthesized by or in: i) the matrix or the medium or environment located outside at least one eukaryotic cell, a prokaryotic cell, or a part of such a cell, or ii) the extracellular matrix.

[0194] 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 mineral iron oxide, the organization of the nanoparticles, for example into chains or aggregates, involves or is due to a living organism.

[0195] The invention also relates to the nanoparticles to be used, wherein the nanoparticles are magnetosomes synthesized by, derived from, extracted from, or isolated from magnetotactic bacteria.

[0196] In one embodiment of the invention, the magnetosome is synthesized by, produced by, derived from, extracted from, isolated from magnetotactic bacteria.

[0197] In one embodiment of the invention, the magnetotactic bacteria are selected from the group consisting of: the AMB-1 strain of Magnetospirillum ma-gneticum, the MC-1 strain of Coccus magnetotactica, three strains MV-1, MV-2 and MV-4 of facultative anaerobic vibrions, the MS-1 strain of Magnetospirillum magne-totacticum, the MSR-1 strain of Magnetospirillum gryphiswaldense, a facultative anaerobic spirillum ma-gnetotacticum, the MGT-1 strain of Magnetospirillum magneticum, and an obligate anaerobe, and Desulfovibrio magneticus RS-1.

[0198] In one embodiment of the invention, a magnetotactic bacterium is defined as a bacterium capable of synthesizing magnetosomes, in which these magnetosomes are preferably characterized by at least one of the following properties: i) they are produced intracellularly, ii) they are magnetic, iii) they comprise a mineral, iv) their nucleus is preferably composed of a metal oxide such as iron oxide, v) their nucleus is surrounded by biological material such as lipids, proteins, endotoxins, which can preferably be eliminated, vi) they are arranged in chains, vii) they produce heat under the application of an alternating magnetic field.

[0199] In one embodiment of the invention, the magnetosomes possess one or more properties common to nanoparticles such as at least one magnetic, size, composition, chain arrangement, charge, core, mineral, coating or crystallinity property.

[0200] In one embodiment of the invention, the magnetosomes comprise the mineral portion synthesized by magnetotactic bacteria, that is, preferably the crystallized iron oxide produced by these bacteria. In this case, the magnetosomes or the mineral portions of the magnetosomes preferentially do not comprise proteins, lipids, endotoxins, or biological materials comprising carbon, or do not comprise more than, 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.

[0201] The invention also relates to the composition according to the invention, wherein CiFRPC and / or C2Frpc is / are one / the photosensitizer(s), preferably selected from the group consisting of: 1) acridine, such as acridine orange, acridine yellow, 2) ALA (5-aminolevulinic acid), 3) aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) aminolevulinic acid, delta-aminolevulinic acid, 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) Mono-ring benzo-porphyrin derivative A, 18) Monoaspartyl chlorin(e6), 19) Talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrin, 23) Chlorine e6 porphrin, 24) Coal tar and derivatives, 25) Contraceptives,oral and estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview, 29) Dyes such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) , Squarains, 33) BODIPY dyes, 34) Phenalenones, 35) Benzophenoxazinium dyes, 36) Erythrosine, 37) Flavines, 38) Foscan, 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(l-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) Blue methylene, 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) Phatalocyanin-based dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phtalocyanin such as liposomal ZnPC, 65) Chloroaluminum sulfonated phthalocyanine (CASP), 66) Silicon phthalocyanine (PC4),67) RLP068, 68) Sodium porfimer, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-tetrakis(l-methylpyridinium-4-yl) porphyrin tosylate, 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 etio-purpurin, 84) Titanium dioxide, 85) Toldudine Blue O, 86) Transition metal compounds such as ruthenium(II), polypyridyl complexes, ruthenium, rhodium, Rh(II)-Rh(II) bridged cyclometallic dimer compounds, platinum(II), gold(III), 87) Verteporfin, 88) Vulcan-based compounds such as aminovulinic acid, aminovulinic acid, 89) WST11, and 90) Xanthene.

[0202] The invention also relates to the composition according to the invention, wherein CiFRPC and / or C2Frpc is / are one / one sonosensitizer(s), preferably selected 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 multi-functional protein, 6) Au Nanomaterial, 7) gold, 8) Au-MnO nanomaterial, 9) manganese oxide, 10) Antineoplastic drugs, 11) NSAIDs, 12) non-steroidal anti-inflammatory drug, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) TiO2 doped with 17) Au, 18) carbon-based nanomaterial, 19) carbon nanotube, 20) chlorine, 21) Ce6, 22) PTX, Paclitaxel, 23) drug or chemotherapeutic compound, 24) infrared dye or IR783, 25) curcumin, 26) cyanine or Cu-Cyanine, 27) DHMS, 28) dimethyl sulfide, 29) docetaxel, 30) drug or chemotherapeutic compound, 31) DOX / Mn-TPPS@RBCS, 32) doxorubicin,32) manganese, 33) globule, 34) red blood cell, 35) polymer, 36) elastomer, 37) erythosine or erythosine B, 38) FA or FA-OI or FA-OI NP or acid, folic acid, 39) F3-PLGA@MB / Gd NPs, 40) poly(lactic-co-glycolic acid), 41) gadolinium, 42) Fe-TiO2 or titanium dioxide, 43) Fe-VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49) hydroxychloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysaffron 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) HSA-wrapped metal-porphyrin complex, 66) Albumin, 67) MnWOx, 68) MnWOx-PEG, 69) PEG, 70) Bimetallic oxide, 71) Mn(III)-HFs, 72) Manganese, Hemoporphine, 73) Nano-beads, 74) Noble metal nanomaterial, 75) O1 NP or oxygen indyocyanine, 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, isomer or phosphate of tris(l-chloro-2-propyl), 87) TPI or thermoplastic polyimide or thermoplastic polymer, 88) TPZ or Tirapazamine, 89) Transition metal oxide, 90) Nanoparticle or Janus nanoparticle, and 91) Xanthones.

[0203] The invention also relates to the composition according to the invention, wherein Cifrpc and / or C2frpc is / are radiosensitizers, preferably selected 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 doped lanthanide 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 nanotubes, quantum dots, nanorads, triflates or metal oxides, 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) Membrane-active agent, 46) Mitomycin-C or Mitomycin, 47) Motexafin, 48) , NBTXR3, 49) Oligonucleotide, 50) Paclitaxel, 51) Papaverine or pa-paverine 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) Sulphur group suppressors, 62) SYM004, 63) Texa-phyrins, 64) TH-302, and 65) Tirapazamine.

[0204] In one embodiment of the invention, CiFRPC and C2FRPC act in synergy, that is to say that the photosensitizing or sonosensitizing or radiosensitizing force of the nanoparticle comprising CiFRPC and C2FRPC is greater than the photosensitizing or sonosensitizing or radiosensitizing force of the nanoparticle comprising only CiFRPC or C2FRPC.

[0205] In some cases, the photosensitizing or sonosensitizing or radiosensitizing strength is the concentration or quantity of radical species, preferably free radical species, produced or captured by at least one photosensitizer, radiosensitizer, sonosensitizer, free radical capture or production center, or composition, or is proportional to the strength or intensity of acoustic background, ultrasound, radiation, light, electromagnetic radiation, preferably applied to the photosensitizer, radiosensitizer, sonosensitizer, free radical capture or production center, or composition.

[0206] In one embodiment of the invention, CiFRPC and C2FRPC act in an antisynergistic manner, that is to say, the photosensitizing or sonosensitizing or radiosensitizing force of the nanoparticle comprising CiFRPC and C2FRPC is less than the photosensitizing or sonosensitizing or radiosensitizing force of the nanoparticle comprising only CiFRPC or C2FRPC.

[0207] In one embodiment of the invention, the composition is introduced or administered to or into the body part, preferably of an animal or a human. Preferably, after its introduction or administration, the nanoparticle first comprises CiFRPC and C2FRPC for a time t1 and then comprises CiFRPC for a time t2, where t2 follows t1.

[0208] In some cases, tl 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).

[0209] In some other cases, tl and / or t2 may be shorter than 1010, 105, 103, 10, 0, 1.5, 10-1 or 10-3 second(s) or minute(s).

[0210] In some cases, t2 can be separated from tl by more than del0-10,10-5,10-3,10-1,0,1,5,10 or 103 second(s) or minute(s).

[0211] In some other cases, t2 can be separated from tl by less than 1010,105,103,10, 0, 1.5, 10-1 or 10-3 second(s) or minute(s).

[0212] The invention also relates to the composition according to the invention, where the composition is or includes: i) a medical device, ii) a medicinal product, 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 lyophilized suspension or composition or preparation.

[0213] 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.

[0214] In some cases, the disease may be due to the malfunction of an organ or part of the body preferentially in an individual.

[0215] In some cases, treatment may be the therapy and / or diagnosis of a disease or a cosmetic treatment.

[0216] In some 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, the cell death possibly occurring by apoptosis or necrosis.

[0217] The invention also relates to nanoparticles for use according to the invention, the disease is chosen from the group consisting of: a disease associated with cell proliferation different from cell proliferation in a healthy individual, a disease associated with the presence of pathological cells in the body part, a disease associated with the presence of a pathological site in an individual or a body part, a disease or disorder or dysfunction of the body part, a disease associated with the presence of radio-resistant or acoustic cells, 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, an eye disease or disorder, an intestinal disease, a communication disorder, a genetic disorder, a neurological disorder, a voice disorder,a vulvovaginal disorder, a liver disorder, a heart disorder, a heating disorder, a mood disorder, anemia, preferably iron deficiency anemia, and a personality disorder.

[0218] In some cases, the disease or disorder may be the disease or disorder of the individual or of the body part or belonging to the individual, or the disease or disorder from which the individual suffers.

[0219] In one embodiment of the invention, cancer or tumor is chosen from the group consisting of: cancer of an organ, blood cancer, cancer of a system of a living organism, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colon / rectal cancer, endometrial cancer, esophageal cancer, eye cancer, gallbladder cancer, heart cancer, kidney cancer, laryngeal and hypopharyngeal cancer, leukemia, liver cancer, lung cancer, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small bowel cancer, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, uterine cancer uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's tumor, Wilms macroglobulinemia, Castleman's disease, Ewing family tumor, gastrointestinal carcinoid tumor, gastrointestinal stromal tumorMyelodysplastic syndrome, pituitary tumor, and cancerous diseases such as gestational trophoblastic disease, Hodgkin's disease, Kaposi's sarcoma, malignant mesothelioma, and multiple myeloma.

[0220] The invention also relates to a method for the treatment of anemia or anemic disease or nanoparticles, in particular magnetosomes, for use in the treatment of anemic disease, preferably iron anemic disease, in which the magnetosomes are administered to the part of the body of an individual, preferably to reduce or stop the anemia.

[0221] The invention also relates to a method for the treatment of an anemic disease, wherein this disease is selected from the group consisting of: iron or metal deficiency anemia, vitamin deficiency anemia, chronic disease anemia, aplastic anemia, anemia associated with bone marrow disease, hemolytic anemia, sickle cell disease, thalassemia, pernicious anemia, Fanconi anemia, sideroblastic anemia, congenital dyserythropoietic anemia (CDA), Diamond Blackfan anemia and megaloblastic anemia.

[0222] 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.

[0223] The invention also relates to a method for manufacturing the composition according to the invention, which comprises at least one of the following steps: - Step 1 of amplification of 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; an iron source 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, FeC13, and their combinations, at a concentration preferably between 1 nM and 2.10-3 Mol / L; a nitrogen source preferably chosen from the group consisting of: at least one compound comprising at least one nitrogen atom, a nitrate salt, nitrogen gas, ammonium, ammonia, an ammonium salt, urea, an amino acid, ammonia gas and combinations thereof, at a concentration preferably between 1 nM and 4 Mol / L; -an oxygen source chosen preferably from the group consisting of: at least one compound comprising at least one oxygen atom, oxygen or air or compressed air, preferably in gaseous form, the oxygen source being in some cases bubbled or introduced into the growth medium, at a gas flow rate which is preferably between 5 mL of gas per minute and 50000 mL of gas per minute; a phosphate source 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 potassium source 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 consisting preferably of at least one compound comprising at least one atom of sulfur or sulfate, at a concentration preferably between 1 nM and 4.10-1 Mol / L; a manganese source consisting preferably 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 vitamin source 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-aminobenzoic acid, pyridoxine HCl, riboflavin, thiamine, thiamine HCl and their derivatives and combinations, at a concentration preferably between 1 nM and 10⁻⁴ Mol / L, and a calcium source 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 magnetosomes with CiFRPC or another metal other than iron, preferably zinc or aluminum, for example a zinc source, 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 comprising preferably a mass percentage of organic matter from magnetotactic bacteria of less than 100, 50 or 1%, - Step 3 of coating the magnetosome minerals with a coating material comprising preferably the C2Frpc compound by mixing the magnetosome minerals with the coating material, wherein 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 amount or mass of the magnetosome minerals and the amount or mass of the coating material, preferably of compound D, which is preferably adjusted or varied or greater than 1, - step 4, which consists of adding at least one cryoprotectant to the coated magnetosome minerals obtained at the end of step 3, - Step 5 of freeze-drying, dehydration, 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 another liquid.

[0224] 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 pM, preferably 2 and 50 pM, more preferably 5 and 20 pM.

[0225] The invention also relates to the process according to the invention, wherein the nanoparticles manufactured according to the process comprise an amount of metal other than iron, preferably zinc or aluminum, which is incorporated in or contained within the metal core is: i) between 0.1 and 100 mg of metal other than iron contained in the core per gram of iron contained in the core, ii) preferably between 0.5 and 10 mg of metal other than iron contained in the core per gram of iron included in the nucleus, iii) more preferably between 1 and 5 mg of metal other than iron included in the nucleus per gram of iron included in the nucleus.

[0226] In one embodiment of the present invention, the nanoparticle is or belongs to or is included in the group of nanoparticles selected from: a nanosphere, a nanocapsule, a dendrimer, a carbon nanotube, a lipid / solid nanoparticle, a lipid- or protein- or DNA- or RNA-based nanoparticle, 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 metallic 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.

[0227] In another embodiment of the present invention, the nanoparticle is not, 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 carbon nanotube, a lipid / solid nanoparticle, a lipid- or protein- or DNA- or RNA-based nanoparticle, 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 metallic 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.

[0228] In some cases, the nanoparticle may be in liquid, gaseous or solid form, preferably before, during or after its presence or administration in the body part.

[0229] In other cases, the nanoparticle may not be in one or two of the liquid, gaseous or solid states, preferably before, during or after its presence or administration in the body part.

[0230] In other cases, nanoparticles may be considered as or included in a ferrofluid, a chemical or biological ferrofluid, where chemical and biological ferrofluids are iron-containing fluids, preferably forming nanoparticles, which are produced by chemical or biological synthesis, respectively.

[0231] In other cases, the ferrofluid or the assembly of nanoparticles may understanding nanoparticles and an excipient, solvent, matrix, gel, which preferentially allows the administration of nanoparticles to the individual or body part.

[0232] In other cases, the nanoparticle may comprise a synthetic material and / or a biological material and / or an inorganic material and / or an organic material.

[0233] In one embodiment of the invention, the nanoparticle(s) is or refers to: 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.

[0234] In one embodiment of the invention, the nanoparticle(s) or the nanoparticle(s) represent or are an assembly or a suspension or a 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 a temperature increase, a radical or reactive species, or the dissociation of a compound from the nanoparticles.

[0235] In another embodiment of the invention, the nanoparticle or nanoparticles represent or are an assembly or suspension or 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 contained 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 contained in the nanoparticle(s) per cm3 or mg of iron contained in the nanoparticle(s) per cm3 of the body part. In some cases, an assembly or suspension or composition of nanoparticles comprising a small number of nanoparticle(s) may be used to prevent toxicity.

[0236] In one embodiment of the invention, the nanoparticle or nanoparticles, or the set of nanoparticles, may represent or be the region, also referred to as the 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 referred to as the nano- particles.

[0237] In some cases, the nanoparticle region may be the volume occupied by a set of nanoparticles in the part of the body, where the nanoparticles are preferentially separated by less than 109, 106, 103 or 10 nm.

[0238] 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 administration of nanoparticles to or into the body part.

[0239] 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.

[0240] In some cases, the distribution of separation distances between nanoparticles may highlight the presence of a minority of nanoparticles, i.e. 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, i.e. separation distances preferentially less than 10, 10, 10 or 0 nm, or large separation distances, i.e. separation distances preferentially greater than 10, 10 or 10 nm. i.e. separation distances preferentially less than 109, 106, 103 or 10 nm, or with large separation distances, i.e. separation distances preferentially greater than 109, 106, 103 or 10 nm. In this case, the presence of this minority of nanoparticles is preferentially not taken into account when estimating the average or maximum separation distance between nanoparticles.

[0241] The invention also relates to nanoparticles for use according to the invention, in which the nanoparticles are crystallized, metallic or magnetic.

[0242] 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 preferably by electron microscopy.

[0243] In one embodiment of the invention, the nanoparticles are metallic. In this case, they contain at least 1, 10, 10³, 10⁵, or 10⁹ metallic atom(s) or contain at least 1, 10, 50, 75, or 90% metallic atoms, this percentage being the ratio of the number or mass of metallic atoms in the nanoparticle divided by the total number or mass of all atoms in the nanoparticle. The nanoparticles, preferably metal oxide nanoparticles, may also contain at least 1, 10, 10³, 10⁵, or 10⁹ oxygen atom(s), or contain at least 1, 10, 50, 75, or 90% oxygen atoms, where this percentage may 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.

[0244] Dans un autre mode de realization de l'invention, le métal ou l'atom métallique est choisi dans la liste constitutee par : Lithium, Beryllium, Sodium, Magnesium, Aluminium, Potassium, Calcium, Scandium, Titanium, Vanadium, Chrome, Manganese, Fer, Cobalt, Nickel, Cuivre, Zinc, Gallium, Rubidium, Strontium, Yttrium, Zirconium, Niobium, Molybdenum, Technetium, Ruthenium, Rhodium, Palladium, Argent, Cadmium, Indium, Etain, Cesium, Barium, Lanthanum, Cerium, Praseodymium, Neodymium, Promethium, Samarium, Europium, Gadolinium, Terbium, Dysprosium, Holmium, Erbium, Thulium, Ytterbium, Lutetium, Hafnium, Tantalum, Tungstène, 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, Copemicium,Nihonium, Flerovium, Moscovium, and Livermorium or Livermorium atom. ,

[0245] In another embodiment of the invention, the nanoparticle contains fewer than 1, 10, 10³, 10⁵, or 10⁹ metal atoms, or contains fewer than 1, 10, 50, 75, or 90% metal atoms, where this percentage may 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 fewer than 1, 10, 10³, 10⁵, or 10⁹ oxygen atoms, or contain fewer than 1, 10, 50, 75, or 90% oxygen atoms, where this percentage may be the ratio of the number or mass of oxygen atoms in the nanoparticle divided by the total number or mass of all atoms in the nanoparticle.

[0246] 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 preferably chosen from the group consisting of a diamagnetic, superparamagnetic, paramagnetic, ferromagnetic and ferrimagnetic behavior or property.

[0247] 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.

[0248] 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.

[0249] In 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.

[0250] In one embodiment of the invention, the nanoparticles have or are characterized by at least one of the following properties: i) the presence of a nucleus, preferably magnetic, preferably mineral, preferably composed of a metallic 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 nucleus and preferably prevents the aggregation of the nanoparticles, preferably allowing the administration of the nanoparticles into an organism or into a part of the body or stabilizing the nucleus of the nanoparticles, the thickness of the coating being preferably 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 of remanent to 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 above 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K, viii) crystallinity, i.e. nanoparticles, preferably measured or observed at a temperature above 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K.that is, nanoparticles preferably possessing at least 1, 2, 5, 10, or 100 crystal plane(s), observable or measured preferably by electron microscopy, ix) the presence of a single domain, x) a size greater than 0.1, 0.5, 1.5, 10, 15, 20, 25, 30, 50, 60, 70, 80, 100, 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, this which preferably means that the nanoparticles have an endotoxin concentration of less than 1020, 10000, 1000, 100, 50, 10, 5, 2 or 1 EU (endotoxin unit) per mg of nanoparticle or per mg of iron contained in the nanoparticles, or which means that the nanoparticles do not trigger fever or an increase in whole body temperature greater than 100, 50, 6.6, 5, 3, 2 or 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 from the living synthesizing organism, xv) the presence of more than 99, 95, 80, 70, 60, 50 or 25% of mineral matter from the living synthesizing 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 an intensity preferably greater than 0.1, 1, 10 or 100 mT, and / or of a frequency greater than 1, 10, 100 or 1000 KHz, alternatively measured preferably under the application of acoustic background, alternatively under the application of radiation such as acoustic electromagnetic radiation, or light.

[0251] 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 remanent magnetization and saturation 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 below 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% of organic or carbonaceous matter from the synthesizing living organism, vii) the presence of less than 99, 95, 80, 70, 60, 50 or 25% of mineral matter from the synthesizing living organism, or xi) a specific absorption rate (SAR) of less than 1, 10, 1000 or 10⁴ Watts 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 a frequency preferably lower than 1, 10, 100, 103, 105 or 109 KHz, alternatively measured preferably under the application of acoustic background, alternatively under the application of radiation such as acoustic electromagnetic radiation, or light.

[0252] In some cases, the mineral may be the part of the nanoparticle or magnetosome that does not comprise organic matter or that comprises a low percentage by mass of organic matter, preferably less than 100, 99, 50, 20, 10, 5, 1, 10⁻¹ or 10⁻² percent or percentage by mass of organic matter. The mineral is preferably the core of the nanoparticle.

[0253] In some other cases, the mineral may comprise a mass percentage of organic matter greater than 0.10⁻⁵, 10⁻¹, 10⁻², 10⁻¹, or 1 percent or mass percentage of organic matter. This may occur when the purification step fails to remove the organic matter or when the organic matter is added to the mineral after the purification step.

[0254] In some cases, the nanoparticles may be surrounded by a coating. The coating may consist of a synthetic, organic or inorganic material or of a substance comprising a functional group selected 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 carboxymethyl dextran, citric acid, phosphatidylcholine (DOPC), or oleic acid. In some cases, the coating may allow the dispersion of the nanoparticles in a matrix or solvent such as water, preferably without aggregation or sedimentation of the nanoparticles. In some cases, the coating may allow the internalization of the nanoparticles into cells.In other cases, the coating can: i) link two or more nanoparticles together, preferably in a chain, ii) prevent the aggregation of nanoparticles and / or, iii) obtain a uniform distribution of nanoparticles.

[0255] 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 body part or EU per mg of nanoparticle or EU per cm3 of body part per mg of nanoparticle, or ii) induce a temperature increase in the individual or body part of less than 105, 103, 102, 50, 10, 5, 4, 3, 2 or 1°C, preferably above physiological temperature, preferably before, after or without the application of acoustic background or radiation to the nanoparticle.

[0256] In one embodiment of the present invention, the nanoparticle or compound is composed of or comprises a chemical element from the families selected 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.

[0257] In another embodiment of the invention, the nanoparticle or compound is composed of or comprises a chemical element selected from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, cesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, the lanthanides, the 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, co-pemicum, 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.

[0258] In some cases, the nanoparticle or compound may also be composed of or comprise an alloy, mixture, or oxide of that chemical element(s).

[0259] 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 elements, where this percentage may represent the mass or number of these chemical elements included in the nanoparticle or compound divided by the total number or total mass of all the chemical elements included in the nanoparticle or compound or by the total mass of the nanoparticle or compound.

[0260] In some other cases, the nanoparticle or compound may be composed of or comprise 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).

[0261] In 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.

[0262] In one embodiment of the present invention, the nanoparticle or compound is not composed of or does not include at least one chemical element belonging to the family chosen from the group consisting of: metals (alkali metal, alkaline earth metal, transition metals), semi-metals, non-metals (halogen element, noble gas), chalcogen elements, lanthanides, actinides.

[0263] In another embodiment of the invention, the nanoparticle or compound is devoid of or does not comprise at least one chemical element selected from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, cesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, the lanthanides, the 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.

[0264] 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 / these chemical element(s).

[0265] In one embodiment of the invention, a 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 greater remanent magnetization and / or may absorb acoustic background energy or power 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.

[0266] 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.

[0267] 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 assembly of nanoparticles has a well-defined, preferably narrow, size distribution.

[0268] 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 size distribution of the nanoparticles may be desirable to prevent aggregation, or to promote chain organization of the nanoparticles.

[0269] In yet another embodiment of the invention, the nanoparticle size distribution is greater than 1000, 100, 75, 50, 25, 10, 5, 2 or 1 nm. A wide nanoparticle size distribution can in some cases allow for faster removal of the nanoparticles.

[0270] 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 large surface charge at low pH when surrounded by a coating that allows it to achieve such a charge without being destroyed.

[0271] 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 can have a low surface charge at high pH when it is surrounded by a coating that allows it to achieve such a charge without being destroyed.

[0272] In another embodiment of the invention, the nanoparticle has a surface charge 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 below 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14.

[0273] In another embodiment of the invention, the nanoparticle has a surface charge 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.

[0274] In another embodiment of the invention, the nanoparticle has a weight or mass, preferably expressed in units such as grams (g), kilograms (kg), or milligrams (mg). One gram of nanoparticle may be one gram of metal such as iron contained within the nanoparticle. The mass or weight of the nanoparticle may correspond to the mass or weight of a single nanoparticle or to the mass or weight of an assembly of nanoparticles.

[0275] 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 desirable to increase the amount of acoustic wave energy absorbed by the nanoparticle.

[0276] 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 mass of nanoparticles may be desirable to prevent or minimize nanoparticle toxicity.

[0277] 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.

[0278] 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 some cases, short chains of nanoparticles may be desired or obtained, for example to promote the internalization of nanoparticles into cells or after partial or total destruction of long chains.

[0279] 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 each chain 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 the aggregation of the nanoparticles or to allow a uniform distribution of the nanoparticles.

[0280] 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 chain between 2 and 105, 2 and 103, 2 and 102, or 2 and 50.

[0281] 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).

[0282] Preferably, when the nanoparticles are biologically synthesized, the nanoparticles can be arranged in a chain: i) inside the organism that synthesizes them, also referred to as the synthesizing living organism, or ii) outside that organism. Preferably, the nanoparticles are arranged in a chain after or before their extraction or isolation from that organism.

[0283] In one embodiment of the invention, the nanoparticles are not arranged in chains.

[0284] In another embodiment of the invention, the nanoparticles are chemically synthesized or are not synthesized by a living organism when fewer than 1, 2, 5, 10, or 100 steps of their production, such as the crystallization of iron oxide, the stabilization of mineral iron oxide, or the organization of the nanoparticles, involve or are 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, which involve chemical reactions occurring without the involvement of living organisms, or parts of living organisms such as DNA, RNA, proteins, enzymes, or lipids.

[0285] In another embodiment of the invention, a chemical synthesis may beused to produce a chemical substance or compound that mimics, copies, or reproduces the compartment, organelle, or other biological material, this chemical synthesis or chemical substance being usable or leading to the production of nanoparticles. In some cases, the compartment, organelle, or other biological material may be a lysosome, an endosome, a vesicle, preferably biological material that has the capacity or function either to dissolve or transform crystalline iron into free iron, or to transform free iron into crystalline iron. In some cases, this transformation is partial and preferentially results in the destruction or formation of a partially crystalline assembly of iron atoms or ions, or preferentially results in a mixture of crystalline and non-crystalline iron.In some cases, crystallized iron can be defined as an assembly of iron atoms or ions that leads to the presence of crystallographic planes, preferentially observable using a technique such as transmission or scanning electron microscopy as a characterization method, and free iron can be preferentially defined as an assembly of iron atoms or ions that does not lead to the presence of crystallographic planes, preferentially highlighted by the absence of diffraction patterns, using, for example, transmission or scanning electron microscopy as a characterization method.

[0286] The invention also relates to nanoparticles for use, wherein 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 according to a method similar to that of these nanoparticles but having improved or additional properties such as a chain arrangement.

[0287] In some cases, chemical analogues of magnetosomes can be chemically synthesized and / or are not synthesized by magnetotactic bacteria.

[0288] In some cases, 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 a 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 chain.

[0289] In one embodiment of the invention, nanoparticles or magnetosomes are purified to remove more than 10, 50, or 90 percent by mass of endotoxins and / or other biological material such as proteins or lipids from the living synthesizing organism or magnetotactic bacteria. In some other cases, the nanoparticles or magnetosomes are purified to remove less than 100, 99.9, 99, 95, or 90 percent by mass of endotoxins and / or other biological material. This purification step preferentially yields 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 amounts of endotoxins, biological material, proteins, or lipids before and after the purification step, respectively.

[0290] In some cases, the purification step may consist of using a method or detergent(s) such as NaOH and / or KOH, which is / are preferably mixed with the living synthesizing organism or magnetotactic bacteria or bacterial debris, preferably to remove organic matter or separate organic matter from inorganic matter contained in nanoparticles or magnetosomes and preferably then be able to harvest the mineral nanoparticle or magnetosome, preferably contained in nanoparticles or magnetosomes.

[0291] In some cases, the purified nanoparticles or magnetosomes are nanoparticle or magnetosome minerals.

[0292] In one embodiment of the invention, the nanoparticles according to the invention are drugs, medical devices, cosmetic products, biological products, products used for research purposes, or products used to determine the properties of biological samples.

[0293] The invention relates to a method for storing the composition according to the invention, which comprises at least one of the following steps:

[0294] - Step 1: Choose or prepare the composition in the form of a liquid suspension,

[0295] - step 2: Lyophilization, desiccation, dehydration of the composition, or eli water content of the composition,

[0296] - Step 3: Storage of the composition in powder form, preferably during more than 1 second, 3 months or 1 year. And

[0297] - Step 4: Suspension or resuspension of the composition, of preference in water.

[0298] EXAMPLE 1: Lyophilized citric acid- and carboxy-methyl-dextran coated magnetosome minerals in the presence of cryoprotectants for long-term storage and sustained anti-tumor activity.

[0299] We report a method for formulating iron oxide nanoparticles na- The process involves producing magnetosomes, which are produced by amplifying magnetotactic bacteria in non-toxic growth media, extracting these nanoparticles from the magnetotactic bacteria under alkaline lysis, and purifying them by heating above 400 °C. This yields pure, non-pyrogenic, uncoated mineral magnetosomes (M-C), which are then coated with biocompatible citric acid or carboxymethyl dextran compounds to produce stable M-CA and M-CMD. The final formulation steps 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, and lyophilizing these mixtures to obtain NP powders of (M-CMD)f and (M-CA)f that exhibit long-term stability, preferably for at least 6 months, retaining their pre-lyophilization properties, i.e., their stability in suspension, their chain arrangement,their carbon content, as well as their surface charge and surface chemical groups, in addition, we have established that (M-CMD)f and (M-CA)f are isotonic, preferably with osmolalities between 275 and 290 mosm / kg H2 O when reconstituting NPs in water, fully biocompatible, i.e. sterile, non-pyrogenic, non-cytotoxic to 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 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, complete biocompatibility, and the potential to destroy tumor cells under hyperthermia.

[0300] Among the various types of nanoparticles (NPs), iron oxide-based IONPs exhibit one of the highest levels of biocompatibility, thus enabling their use in medical applications, particularly cancer treatment. Chemically synthesized IONPs often suffer from a number of drawbacks, such as the use of toxic compounds in their manufacturing process, crystalline structure, non-uniform shapes, and / or small sizes with low magnetization. To overcome these drawbacks, a biological synthesis route has been developed, in which IONPs, called magnetosomes, are produced intracellularly by Gram-negative magnetotactic bacteria (MTBs).MTBs have fine-tuned the properties of magnetosomes over millions of years through a Darwinian process, resulting in an optimized magnetic guidance system, also known as magnetotaxis, in which the magnetosome's magnetic moment aligns parallel to the Earth's magnetic field. This allows MTBs to orient themselves in this direction, thus facilitating their search for an optimal living environment. Magnetosomes are therefore composed of well-crystallized cuboctahedral nanocrystals. Magnetosome molecules (Fe3O4) are surrounded by a stabilizing phospholipid bilayer derived from MTB. Individually, they range in size from 35 to 120 nm, giving them ferrimagnetic properties. At the larger scale of the magnetosome assembly, they form chains, preventing agglomeration. While these characteristics have led to superior antitumor efficacy for magnetosomes compared to their chemical counterparts, these results were obtained with magnetosomes in unfinished formulations.

[0301] Part of our work involves improving the magnetosome manufacturing process to make these nanoparticles injectable into humans. To this end, toxic CMR or animal-derived compounds are first removed from the 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 uncoated M-magnetosomes, which are pyrogen-free but prone to agglomeration / sedimentation due to strong magnetic dipolar interactions. Third, the uncoated M-magnetosomes are coated with biocompatible compounds via coordination bonds between a coating material, i.e., citric acid (CA) or carboxymethyl dextran (CMD), and iron cations on the mineral surface, to yield 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 resulting mixtures to transform them into a powder of NP, called (M-CA)f and (M-CMD)f, which can be stored for a long period without losing its properties.

[0302] To improve long-term storage stability and prevent disassembly of the magnetosome mineral coating, it is preferable to store coated magnetosome minerals in powder form rather than as a liquid suspension. To remove water from M-CA and M-CMD, we lyophilized these nanoparticles in the presence of various cryoprotectants. The cryoprotective action of these agents likely relies on the creation of an amorphous glassy matrix that immobilizes the nanoparticles during freezing, thus preventing the crystallized ice produced during this step from damaging the coating. Since the effectiveness of cryoprotection likely depends on the type and concentration of the cryoprotectant used in a formulation, we tested various cryoprotectants—glucose, mannitol, PEG 4000, sorbitol, sucrose, and trehalose—at different concentrations, which we added to M-CA and M-CMD.We then determined the optimized conditions for obtaining stable and isotonic (M-CA)f and (M-CMD)f products, which can be stored as a powder and resuspended in water, while retaining their physicochemical properties, i.e. the thickness / composition of their coating or the arrangement of the magnetosome chain, their bio. compatibility, i.e. their sterility, their absence of pyrosis and their resistance to corrosion. i.e. their sterility, their non-pyrogenicity, their non-cytotoxicity towards healthy cells, as well as their ability to destroy PC3-luc tumor cells under hyperthermic conditions of 41-46 °c triggered by the application of low intensity ultrasound or alternating magnetic field.

[0303] MATERIALS AND METHODS:

[0304] Materials: The Magnetospirillum gryphiswaldense strain MSR-1 (DSM 6361) was purchased from the Deutsche Sammlung von Mikro-organismen und Zellkulturen (Brunswick, Germany). The PC3 human prostate adenocarcinoma tumor cell line Pc3-luc was purchased from PerkinElmer. The 3T3 mouse fibroblast cell line was purchased from the American Type Culture Collection (ATCC® CCL-163™). The V79-4 male Chinese hamster lung cell line was purchased from ATCC® (CCL-93). The L929 mouse fibroblast cell line 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 (HNO3), and Triton X-100 were purchased from Thermo Fisher Scientific. Dulbecco's modified eagle medium (DMEM) with / without phenol red, fetal bovine serum (FBS), penicillin-streptomycin, HEPES buffer solution, 0.25% trypsin-EDTA solution, and trypan blue solution were purchased from Gibco. Resazurin dye was purchased from Invitrogen.

[0305] Culture of MSR-1 magnetotactic bacteria: In brief, MSR-1 bacteria were first amplified in two pre-culture (PC) steps with iron-deficient pre-culture medium, and then cultured in the final step under conditions appropriate to promote magnetosome synthesis. During PC 1, 300 µl of MSR-1 cryo-stock were incubated in 50 ml of pre-culture medium at 29.5°C for 6 days under static conditions followed by one day of shaking at 110 rpm. Subsequently, PC2 was performed by transferring all cells into a 15 L bioreactor (Applikon) containing 6 L of pre-growth medium and shaking at 200 rpm and 29.5°C for 3 days. During the final culture stage, the PC 2 cells were transferred into a 40 L bioreactor (Applikon) filled with 26 L of growth medium.Culture conditions such as oxygen concentration, temperature and stirring speed were maintained between 0.1 and 1%, at 29.5°C and under 200 rpm throughout the entire period. The pH was maintained at 6.9 by the automatic addition of an acidic medium (pH 3), referred to as 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 (OD565) using a UV-visible spectrophotometer. Magnetotactic bacteria MSR-1 (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) comprise: i) sodium lactate (preferably 2.6 g / L in PGM, 1.3 g / 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), mineral elixir (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 comprises 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 pentothenate, 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, and 0.4 g / L thiamine HCl. The first 10 days consist of two pre-culture steps used to amplify the bacteria in a pre-culture medium devoid of the iron source. The third step lasts approximately 9 days.This method promotes intracellular magnetosome synthesis by culturing MSR-1 bacteria under optimized microaerophilic conditions and progressively adding iron ion sources via a fed-batch solution. At the end of the bacterial culture, 32 L of the bacterial culture was concentrated to reduce the liquid volume using a tangential flow filtration column. The MSR-1 concentrate (approximately 5 L in total) was then stored at -80°C for subsequent magnetosome production steps.

[0306] Extraction and purification of magnetosomes: Briefly, concentrated MSR-1 cells were diluted to an OD565 value of 20 and then lysed in 2 m KOH at 80°C for 1 h under mechanical stirring at 150 rpm. Using magnetic selection, the magnetosome (MgC) chains were collected and washed twice with 1 x phosphate-buffered saline (PBS) followed by three washes with deionized water. After being concentrated in 50 ml conical tubes, the MgC 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 1) to convert them into powder. Then, 100 mg of this powder were 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 the organic matter from the post-extracted magnetosomes and obtain magnetosome minerals or "uncoated magnetosomes" (M-uncoated).

[0307] Coating of M-Uncoated with citric acid (CA) and carboxymethyl dextran (CMD): The coating process was carried out under aseptic conditions, using a biosafety cabinet and pyrogen-free materials. The CA and CMD solutions were prepared in pyrogen-free water at 25 mg / ml and 150 mg / ml, respectively. These solutions were then filtered through a 0.22 µm polyethersulfone filter for sterilization. Next, 200 ml of each solution (CA or CMD) was added to an 11 ml 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 at room temperature, using a probe sonicator (PS) (Branson, digital sonifier s-250d) with 25 mm diameter tips. After the short 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 sonicated again for 1 h using the PS (Branson, digital sonifier s-250d) with the same parameters as those described previously 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 from the supernatant.Each coated magnetosome mineral was then resuspended in 10 ml of water to obtain a final concentration of 100 mg / ml of iron and stored at +4°C.

[0308] 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 various cryoprotectant solutions, such as glucose, mannitol, sorbitol, sucrose, and trehalose, to obtain 1 ml samples formulated at 50 mg / ml iron containing 5% or 10% w / v cryoprotectant. Each sample was immediately frozen in liquid nitrogen for 15 min after 30 s of vortexing, then lyophilized at -50°C (storage temperature), 0.003 mbar for 20 h for primary drying (labconco, free zone 70020 2.5 1). Next, secondary drying was initiated for 6 hours by increasing the temperature to 40°C (storage temperature) at a heating rate of 0.3°C / min while maintaining the pressure at 0.003 mbar. Similarly, the M-CA suspension at 100 mg / ml iron was tested with various cryoprotectant solutions to obtain the samples. formulated from 1 ml at 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 Tl; and 2.5%, 3.75%, 5%, 6.25% and 7.5% w / v of cryoprotectant for the sucrose-peg 4000 and sucrose-dextran Tl combinations with sucrose-PEG or dextran mass ratios of 1:1, 1:2, 1:3, 2:1, 2:2, 2:3, 3:1, 3:2 and 3:3. After lyophilization, some of the formulated magnetosome samples were resuspended in 1 ml of sterile water for further characterization. The selected formulations of M-CA and M-CMD with cryoprotectants were designated (M-CA)f and (M-CMD)f, respectively.

[0309] Transmission electron microscopy (TEM): 1 ml of MSR-1 bacteria at the end of culture was diluted to an OD565nm of 1 and then rinsed twice with deionized water by centrifugation at 2400 g for 10 min. The different magnetosome suspensions were diluted to 50 pg / ml of iron. Then, 7 µl of each sample were deposited onto a carbon-coated copper grid (300 mesh from Oxford Instruments). The grid was then left to 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 the image j software, the size of the nanoparticles was estimated by measuring the diameter of approximately 400 randomly selected mineral nuclei in the magnetosome chains within the MSR-1 bacteria.

[0310] Colloidal stability by colorimetric assay: 1 ml of magnetosome suspension was vortexed for 30 s and then a 50 µl sample was immediately taken, called "sample t0". The suspension was then left to stand on the bench for 2 hours, and another 50 µL sample was taken from the upper portion of the liquid, referred to as the "t2h sample." For the conversion to 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 20% v / v H₂O₂ for 15 min to oxidize Fe²⁺ to Fe³⁺ ions. Afterward, the solution was mixed with 880 µL of ultrapure water and then with 50 µL of 2 m KSCN to form the complex between Fe³⁺ and thiocyanate ions, the absorption of which at 476 nm was measured using a spectrophotometer. The iron concentration was then determined using a calibration curve.The stability rate of the magnetosome suspension was calculated as the ratio between the iron concentration of sample t2h and that of sample t0.

[0311] Osmolality measurement: 30 µl of 50 mg / ml iron magnetosome suspension were introduced into a micro-sample tube and then placed on the osmometer plate (Advanced Osmometer, Model 2020) to determine the osmolality of each sample.

[0312] CHNS Elemental Analysis: 5 mg of magnetosome powder were packed in an aluminum capsule and introduced into the CHNS Elemental Analyzer (thermofisher, flash 2000) to determine the carbon and nitrogen content from organic matter in each preparation.

[0313] Fourier transform infrared (FT-IR) spectroscopy: 2 mg of magnetosome powder were deposited onto a germanium crystal plate connected to an FT-IR spectrometer. The tip was lowered until it contacted the crystal and thus the powder sample. The FT-IR spectra were then recorded over a scanning range of 590–4000 cm¹ with a resolution of 4 cm¹.

[0314] Zeta potential measurement: Each magnetosome suspension was diluted to 50 pg / ml in iron with ultrapure water. It was then divided into five 2 ml samples to be prepared at different pH levels ranging from 2 to 10 at 25 °C using HCl and NaOH solutions. 1.5 ml of each sample was then introduced into a 4.5 ml disposable cuvette and measured in a zeta potential assay.

[0315] Residual moisture analysis: 10 mg of lyophilized powder were 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 maintained at 120°C for 1 h to determine the weight loss of the lyophilized powder.

[0316] Sterility Test: Under aseptic conditions, 20 mg of each magnetosome powder were 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 for 14 days. Culture media without nanoparticles (NPs) were considered negative controls. On the last day, 1 mL was taken from each sample and placed against a magnet for 10 min to recover only the medium. Turbidity was then measured with 900 µL of this medium at 600 nm. The remaining 100 µL were incubated on a solid Luria-Bertani agar plate at 25°C or 35°C for 3 days to detect any contaminating colonies. The other tests, namely the growth promotion test and the method suitability test, were also carried out in parallel with the magnetosome sterility test in order to verify the robustness of this test.

[0317] Endotoxin quantification by the LAL (limulus amebocyte lysate) assay: Endotoxin quantification was performed on various magnetosome powders using a pierce chromogenic endotoxin quant kit under aseptic conditions. All materials coming into contact with the NP samples were sterile and pyrogen-free. Before the test, each magnetosome powder was prepared with 40 pg / ml iron in endotoxin-free water and then heated to 70°C for 15 min to denature any residual protein that could interfere with the test. After this, 25 µl of this The suspensions were introduced into a 96-well plate pre-equilibrated at 37°C for at least 10 minutes. While maintaining the plate at 37°C, 25 µl of the reconstituted amebocyte lysate reagent were 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 were added to each well to stop the reaction. The optical density of this mixture was then measured at 405 nm using a microplate spectrophotometer. The endotoxin concentration was determined using the standard E. coli endotoxin curve, which was established concurrently with the samples.

[0318] 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 cm2 culture flask containing 10 ml of appropriate culture medium, i.e. DMEM supplemented with 10% v / v 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 HEPES buffer for L929. After being placed in the vial, the cells were maintained at 37°C in a 5% CO2 incubator. The culture medium was changed twice weekly for each vial. When the cells reached approximately 80% confluence, cell passage was performed.To do this, all the liquid medium was first removed from the flask, and 1 ml of 0.25% trypsin-EDTA was added to collect all the adherent cells in suspension. Then, only 0.5 ml of cell suspension was retained to be mixed with 10 ml of fresh medium in a new 75 cm² culture flask and reincubated at 37°C in the 5% CO₂ incubator. After 3 passages, each cell line was ready for further experiments.

[0319] Cytotoxicity assay. The 3T3, L929, and V-79 cell lines were each seeded onto a sterile 96-well plate (10⁴ cells / well) and then incubated overnight at 37°C in a 5% CO₂ incubator for cell adhesion. After this, all the medium was carefully removed from the plate. 100 µl of lyophilized (M-CA)f or (M-CMD)f suspension, prepared at different concentrations in the appropriate phenol red-free culture medium for each cell line, were then added to each well. In the 3T3, V-79, and L929 cell plates, 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% CO2 incubator. To determine cell viability, 10 µl of resazurin reagent was added to each well and homogenized under shaking at 120 rpm for 10 min.The plates were then incubated at 37°C. incubator at 5% CO2 for approximately 4 h. Once the blue color of the untreated cell wells turned pink, all the 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 might interfere with the assay. Next, 100 µl of the supernatant were transferred to a new 96-well plate for measurement at excitation and emission wavelengths of 530 and 590 nm, respectively, using a fluorescence reader. Cell viability was calculated using the formula: Cell viability (%) = ( ) x 100% where , Ft is the fluorescence intensity of the treated wells, Fb is the fluorescence intensity of the blank measured in empty wells containing 100 µl of medium mixed with 10 µl of resazurine, and Fc is the fluorescence intensity of the control wells measured in untreated cells incubated in 100 µl of medium mixed with 10 µl of resazurine.

[0320] Hyperthermia treatments under AMF (alternating magnetic field) and LIU (low-intensity ultrasound) applications. PC3-luc cells were seeded onto a sterile 96-well plate (3 x 0.4 cells / well) and incubated overnight at 37°C in a 5% CO2 incubator for cell adhesion. Subsequently, all the medium was removed from each well, and the adherent cells were rinsed twice with phenol red-free DMEM (white DMEM). 100 µl of (M-CA)f or (M-CMD)f suspension prepared with 1 mg / ml iron in white DMEM supplemented with 10% v / v FBS, 1% v / v penicillin-streptomycin, and 1% v / v HEPES buffer were added to each well. Next, all wells were reincubated for 3 hours at 37°C in a 5% CO2 incubator.After that, the hyperthermia experiment was divided into three conditions, each containing 3 wells with only Pc3-luc cells, 3 wells with cells incubated with (M-CA)f, and 3 wells with (M-CMD)f. In the first condition, the wells were not exposed to any hyperthermia source.In the second condition, three wells were placed in a polystyrene holder positioned at the center of a copper coil. A 42 mt, 195 kHz AMF was applied to the well for 30 minutes using easy-to-use heating equipment to warm the cells. In the third condition, each of the three wells was immersed in an adapter filled with degassed water to connect to a 3.6 cm diameter planar ultrasonic transducer. This allowed them to be exposed for 30 minutes to an ultrasonic intensity between 0.3 and 1 W / cm², at 1 MHz and in continuous mode, produced by an ultrasonic generator (Primo Therasonic 460). To limit evaporation of the culture medium, each well was capped when exposed to a hyperthermic source. The temperature was monitored over time using a flexible thermocouple probe (physitemp, it-18) inserted into each well and the Dasylab software. The SAR of the sample was calculated using the following equation: 5'A / ? ( WI gjx ( - )°ù Cmedium = 4.2 jg 1 -k1 is the capacity The specific heat of water, Cfe, measured in grams of iron per gram of water, represents the iron concentration in the magnetosome sample, and tA (measured in °C / s) is the slope of the temperature change over time in the magnetosome sample after subtracting the temperature of the control samples. After the hyperthermia treatments, all wells were incubated for 24 hours at 37 °C in a 5% CO2 incubator. Cell viability in each well was then determined using resazurin.

[0321] Quantification of iron in magnetosomes internalized by cells. After collecting all the supernatant for the cell viability assay, PC3-luc cells adhering to the bottom of the wells were gently rinsed 3 times with 300 µl of 1x bp. Each well was then incubated with 50 µl of 0.25% trypsin-EDTA for 1 min at 37 °C and 5% CO2, followed by the introduction of 150 µl of complete DMEM. After this, 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 from 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 were centrifuged at 14100 g for 20 min to remove all the supernatant. The cell pellet was then dissolved overnight in 20 µl of 37% w / v HCl and 145 µl of 70% w / v HNO₃ 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 magnetosome internalized in PC3-luc cells.

[0322] Statistical analyses. Data were analyzed using Graphpad Prism version 8.0. All measurements were taken 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.

[0323] RESULTS AND DISCUSSION.

[0324] Magnetosome-based formulation: summary of previously published initial steps and presentation of additional / complementary steps proposed in this study. This study presents additional aspects concerning the implementation of a magnetosome-based formulation for human injection. The initial 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 sizes The mean magnetosome size was 36.6 (standard deviation 6.4 nm). Step 2 involved extracting magnetosomes from magnetotactic bacteria obtained in Step 1 using bacterial KOH lysis and magnetic separation of the magnetosomes from bacterial debris. This yielded chain-like (Mg-Ch) magnetosomes, where each magnetosome is surrounded by a bacterial inflammatory membrane. These magnetosomes appear to be too immunogenic for human injection without specific treatment. Step 3 involved purifying the Mg-Ch by combustion to remove / denature the bacterial organic matter. This resulted in non-pyrogenic (M-uncoated) magnetosome minerals with a low carbon content of 0.2%, which tend to aggregate due to their bare surface.The uncoated M-series are coated with two biocompatible compounds, namely citric acid (CA) or carboxymethyl dextran (CMD), resulting in non-pyrogenic coated magnetosome minerals (M-CA and M-CMD) reconstituted into chains.

[0325] The objective of this example is to add a few additional 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 objective, M-CA and M-CMD were stored as an anhydrous powder, which can be reconstituted into a NP suspension at any time, as needed. We then verified that the reconstituted M-CA and M-CMD, designated (M-CA)f, (M-CMD)f, (M-CA)fw, (M-CMD)fw for lyophilized M-CA and M-CMD before and after resuspension of the NPs in water, respectively, retained their physicochemical properties and antitumor activity.

[0326] Formulation of M-CA and M-CMD in the presence of a cryoprotectant for long-term storage. Here, we determine the manufacturing conditions for a formulation of injectable magnetosomes exhibiting long-term stability. This stability is evaluated by measuring the percentage of absorption stability, measured at 480 nm over time, of a magnetosome suspension, preferably 50 mg of magnetosomes, for 2 hours after homogenization.The percentages 100% and 0% correspond to stable and unstable suspensions, respectively. We chose to perform stability measurements with 50 mg of magnetosomes because we plan to administer this quantity of magnetosomes into a 1-2 cm diameter human prostate tumor based on our preclinical efficacy data in mice extrapolated to humans. Furthermore, a 2-hour time frame was chosen for stability assessment, which appears to be long enough for a nurse or physician to administer a stable magnetosome suspension into tumors. Since the purified magnetosomes lacking active bacterial organic matter (uncoated M-) were unstable, i.e., their stability percentage was 0%, they were coated with CA and CMD. This yielded M-CA and M-CMD, which are perfectly stable, meaning their stability percentage is 100%, higher than that of uncoated M- and M-gC. When M-CA and M-CMD are lyophilized without a specific compound to preserve their coating, they form NP powders designated as (M-CA)fd and (M-CMD)fd, characterized by the presence of agglomerated magnetosomes. When (M-CA)fd and (M-CMD)fd are resuspended in water after the lyophilization step, they rapidly settle, resulting in a stability percentage of less than 5%.To enable the reconstitution of M-CA and M-CMD into stable aqueous NP suspensions after lyophilization—that is, without damaging the M-CA / M-CMD coating—M-CA and M-CMD at the intended therapeutic dose of 50 mg / ml were mixed with various cryoprotectants, namely glucose, mannitol, sorbitol, sucrose, or trehalose, used at mass percentages of 5% and 10%. After lyophilization, these formulations were resuspended in water to determine their colloidal stability. Considering M-CMD first, only the 5% and 10% sorbitols maintained 100% colloidal stability of the NPs. In addition, the osmolality of the M-CMD suspension increases with the percentage of sorbitol added to the M-CMD suspension, going from 0 mosm / kg H2 O in the absence of cryoprotectant to 280 and 560 mosm / kg H2 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 H₂O), which is acceptable for human injection. As for M-CA, M-CA mixed with 10% glucose, sorbitol, or sucrose achieves 100% colloidal stability. However, these formulations are hypertonic, meaning their osmolality exceeds 300 mosm / kg H₂O. To address this issue, we selected M-CA mixed with 10% sucrose, resulting in an osmolality of 330 mosm / kg H₂O, the closest value to plasma osmolality. We then added either PEG 4000 or Dextran Tl to this mixture to lower the formulation's osmolality to match plasma osmolality. While the use of Dextran Tl stabilized the formulation, it resulted in an osmolality lower than plasma osmolality. Therefore, Dextran Tl was discontinued.In contrast, combining sucrose with PEG 4000 resulted in a 100% stable formulation, whereas adding only 7.5% sucrose or 15% PEG 4000 led to stability of approximately 90% and 50%, respectively. This result suggests a synergistic effect between sucrose and PEG 4000, which appear to be mutually supportive; that is, PEG 4000 could effectively protect the nanoparticles (NPs) during freezing by encapsulating them in a glassy matrix, and amorphous sucrose could serve as a "water replacement" reservoir, allowing the formation of hydrogen bonds with PEG 4000. Citric acid is applied to the surface of the nanoparticles (NPs), thus preserving the integrity of the coating against dehydration-induced stress. The optimal combination of sucrose and PEG 4000 is 1.25% PEG 4000, the lowest concentration to prevent the risk of molecular mobility and formulation degradation during storage, which could result from an excessive PEG concentration, and the highest mass percentage of sucrose, 3.15%, to obtain a stable M-CA formulation in suspension. The M-CA formulation with 1.25% PEG 4000 and 3.75% sucrose has an osmolality of 225 mOsm / kg H₂O, which can be adjusted to be isotonic upon reconstitution.

[0327] M-CA / M-CMD retain their physicochemical properties after 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)f and (M-CA)f before cryoprotectant removal and as (M-CMD)wf and (M-CA)wf after cryoprotectant removal, have retained their physicochemical properties, we compared the carbon and nitrogen percentages as well as the FT-IR spectra and surface charges of the formulated magnetosomes with those of the magnetosomes harvested at the different stages preceding formulation, i.e. M-CA, M-CA, M-CA and M-CA, i.e. M-CA, M-CMD, M-gC, and uncoated M-CMD.Regarding the mass percentages of carbon and nitrogen in the magnetosomes at the different formulation stages, they first decrease from %C=22.7% and %N=2.2% in M-gC to %C=0.12% and %N=0.01% in M-uncoated, corresponding to the removal of the magnetosome membrane in M-gC to give M-uncoated. Next, they increase a first time from uncoated M 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 CA and CMD coating to the surface of uncoated M, and a second time from 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., down to %C=1.6% and %N=0.01% in (M-CA)fw and %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 shows vibrational bands of P-O (at 1037 cm⁻¹), C-O (at 1410 cm⁻¹), NH₄⁺ (at 1542 and 1641 cm⁻¹), and OH⁻ (at 3276 cm⁻¹), corresponding to the functional groups present in the phospholipid membrane of the magnetosome. Regarding M-uncoated, its FT-IR spectrum does not indicate the presence of peaks above 1100 cm⁻¹, which is consistent with the elimination of the . most of the organic matter in this sample. The two peaks at 612 and 693 cm⁻¹ are attributed to Fe-O stretch vibrations, which originate from the iron oxide contained within the mineral core of the magnetosome, and are therefore present in all FT-IR spectra of the different types of magnetosomes. Coating uncoated M with CA and CMD leads to a series of peaks in the range of 1000–1300 cm⁻¹ and 1630–1750 cm⁻¹, which correspond to co and c=o stretch vibrations, respectively, attributed to the carboxylate groups of CA and CMD in M-CA and M-CMD. Further in the formulation process, the lyophilized magnetosomes (M-CA)f and (M-CMD)f exhibit intense bands of alcohol and alkane groups, e.g. the CO, OH and CH stretch vibration bands in the range of 970 - 1250 cm⁻¹, 3200 - 3550 cm⁻¹ and 2850 - 3000 cm⁻¹, respectively, corresponding to sucrose and PEG 4000 in (M-CA)f, and to sorbitol in (M-CMD)f.Finally, after a washing step, (M-CA)fw and (M-CMD)fw exhibit 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 the magnetosome coating agents. CHNS and FT-IR measurements suggest that the surface area of ​​the formulated magnetosomes remains unchanged compared to that of the pre-formulated coated magnetosomes. To confirm this conclusion, we measured the surface charges of the formulated magnetosomes and compared them to those of the unformulated magnetosomes. (M-CA)f and (M-CMD)f exhibit 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)f and (M-CMD)f, likely by efficiently protecting the coating layers in M-CA and M-CMD from degradation / removal during lyophilization, and by preventing the cryoprotectant from binding strongly to the nanoparticles (NPs). Furthermore, under conditions of use for human injection, i.e., at the therapeutic dose of 50 mg / ml in iron and at a pH of 6.5, (M-CA)f and (M-CMD)f exhibit surface charges below -30 mV, creating conditions of strong repulsive electrostatic forces in the (M-CA)f / (M-CMD)f suspensions. These forces can prevent NP agglomeration caused by magnetic dipolar interactions and stabilize these NPs in suspension.Electron microscopy measurements performed on formulated magnetosomes washed of their cryoprotectant, i.e., on (M-CA)fw and (M-CMD)fw, show that the formulated magnetosomes retain 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 the desired outcome here, is ensured when the amount of water remaining in the lyophilized product after the lyophilization process is [value missing]. The residual moisture (RM), also called residual moisture (RM), is sufficiently low, typically less than 3% for a pharmaceutical product. The RM content of (M-CA)f and (M-CMD)fa was measured by introducing these nanoparticles (NPs) into a thermal gas analyzer (TGA), heating them at 120°C for 1 hour at a heating rate of 6°C / min, and measuring the weight loss attributed to water evaporation. The RM contents of (M-CA)f and (M-CMD)f are in the range of 1.8–2.4%, which is pharmaceutically acceptable. Furthermore, thanks to this efficient lyophilization process, (M-CA)f and (M-CMD)f can be resuspended in water, preferably after 6 months, where they retain their stability after a six-month storage period.

[0328] Sterility / non-pyrogenicity of formulated magnetosomes. Compared to unformulated magnetosomes, formulated magnetosomes must not only retain their physicochemical properties but also their biocompatibility. To investigate this latter aspect, we first examined the sterility of formulated magnetosomes by introducing (M-CA)f and (M-CMD)f for 14 days into tryptic soy broth (TSB) and thioglycolate fluid medium (FTM) solutions at 30°C and 37°C, respectively. Indeed, these conditions are known to amplify bacterial contaminants when initially present in a test sample, and thus 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 (OD60o) at the end of the incubation time.The OD600 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 OD600 value <0.1, comparable to the OD600 <0.1 of uncoated M- 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 agar plates inoculated with (M-CA)f and (M-CMD)f. To further assess the sterility and non-pyrogenicity of (M-CA)f and (M-CMD)f, the endotoxin concentration of these nanoparticles was measured. (M-CA)f and (M-CMD)f appear to contain a very low concentration of endotoxin of 2-5 EU / mg of iron, comparable to the values ​​measured for uncoated M-.Overall, these results highlight the effectiveness of the magnetosome purification step in removing bacterial contaminants from M-gC, the initial presence of which before specific treatment is revealed by the high DOfflo value of M-gC of 1-2.5 as well as by a high M-gC endotoxin concentration of 50 EU / mg iron. Such behavior can be attributed, on the one hand, to a two-step depyrogenation process, i.e., first by mixing the M-gC with KOH at 80 °C and then by heating the M-gC above 400 °C, and on the other hand, to formulation steps carried out under ascetic conditions, i.e., the coating of the magnetosome minerals with [unclear]. CA or CMD, the addition of cryoprotectant to coated magnetosomes M-CA and M-CMD, the lyophilization of the resulting mixture, are carried out under a sterile hood.

[0329] Non-cytotoxicity of formulated magnetosomes. To examine the cytotoxicity of lyophilized formulated magnetosomes, 100 µl of (M-CA)f and (M-CMD)f suspensions with concentrations ranging from 0.001 to 1 mg of iron NP per ml were contacted with different mammalian cell lines, 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 assay. All cell lines exhibit 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 tested conditions, and that the presence of a cryoprotectant and a lyophilization step in the magnetosome formulation does not result in additional cytotoxicity.

[0330] Effective treatment of hyperthermia using formulated mineral magnetosomes 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. More specifically, we examine whether (M-CA)f and (M-CMD)f effectively destroy prostate tumor cells when 100 µL of (M-CA)f or (M-CMD)f at a concentration of 1 mg 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 an AMF of 42 mt and 195 kHz or to low-intensity ultrasound of 0.3–1 W / cm² and 1 MHz.Such excitation conditions lead to a temperature increase in these mixtures from ambient temperature (22–25°C) to a moderate hyperthermic temperature of 46°C. The temperature increase is significantly faster with ultrasound than with magnetic excitation, resulting in a temperature maintenance of 46°C for almost 30 minutes and slightly less than 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 nanoparticles (NPs) to AMF / LIU after subtracting the initial temperature increase due to AMF / LIU in the absence of NPs.While (M-CA)f and (M-CMD)f exhibit a high SAR value of 229 ±16 W / gFe under AMF excitation, they produce a near-zero SAR value under LIU, highlighting the different nature of the heating with the two excitation sources: heating from the excitation of nanoparticles by AMF or from the absorption of ultrasonic energy by the cells, using AMF or LIU, respectively. Furthermore, the heating properties of the two types of formulated magnetosomes, (M-CA)f and (M-CMD)f, are similar. This indicates that the differences between the two coatings are... In terms of nature / composition, coating thickness, and carbon content, these factors do not significantly affect the heating properties of (M-CA)f and (M-CMD)f. We can conclude that certain contributions to heating, such as the Brownian contribution in the induction mechanism, which may depend on coating properties, are not dominant in the observed heating properties. To more precisely assess the effectiveness of heat treatment in destroying tumor cells, Pc3-luc cells treated as described above were reincubated overnight at 37°C under 5% CO2. 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 is consistent with that observed in healthy mammalian cells. In contrast, in the presence of M-CA / M-CMD and a heating session, cell viability decreases by 30–40% and 60–85% after the application of AMF and LIU, respectively, compared to the two controlled conditions in which cells are either incubated solely with (M-CA)f / (M-CMD)f without heating, or exposed solely to AMF / LIU without exposure to nanoparticles. These results suggest that LIU leads to more efficient cell destruction than AMF, possibly due to the temperature being 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 for destroying prostate tumor cells, M-CMD outperforms M-CA, a behavior we are currently trying to elucidate.Given that NP internalization in tumor cells is often correlated with efficient cell destruction, we estimated the amount of (M-CA)f and (M-CMD)f internalized in PC3-luc cells after treatment similar to that used to measure cell viability. After treatment, we destroyed and dissolved the tumor cells and measured their iron content by ICP-MS. The amounts of NP internalized in PC3-luc tumor cells, Q, were greater for (M-CMD)f than for (M-CA)f under all tested conditions, i.e., with / without AMF / LIU treatment. Q is estimated at 17–20 and 30 pg of NP per cell after AMF / LIU treatment for (M-CA)f and (M-CMD)f, respectively. Compared to (M-CMD)f, (M-CA)f shows lower internalization in 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 lead to a greater affinity of (M-CMD)f for cationic sites in the plasma cell membrane and, consequently, greater cellular internalization, perhaps by pinocytosis, for (M-CMD)f than for (M-CA)f. If we now consider the potential impact of... Regarding the impact of nanoparticles (NPs) on cell viability, we observed that the increased NP internalization between M-CMD and M-CA is either correlated with increased cell death after LIU treatment or not correlated with a change in cell death under AMF application. These behaviors could be explained by the fact that ultrasound acts synergistically with internalized NPs to destroy tumor cells; that is, ultrasound could destroy / inactivate the cell membrane while the NPs could act disruptively within the cells from their intracellular location. In contrast, while AMF is expected to produce local heating at the NP level, it is not expected to act disruptively on the cell membrane.Thus, it is possible that in the presence of internalized NPs, ultrasound acts in a more disruptive way than AMF, resulting in more efficient cell destruction.

[0331]

Claims

1.

2.

3. Demands Composition comprising at least one chain of at least two nanoparticles, wherein each nanoparticle of the chain comprises an iron oxide mineral core surrounded by a coating, wherein the composition further comprises a cryoprotectant, characterized in that the volume occupied by the cryoprotectant in the composition is greater than the volume occupied by at least one chain in the composition by a factor of at least 1, 2, 5, 10 or 10³, and wherein the mass percentage of the cryoprotectant is between 0.5 and 50% and the cryoprotectant is selected from the group consisting of: Acetamide, Acetate, Albumin, Ammonium Acetate, Arginine, Alcohols containing at least one or two hydroxyl groups, Choline Bromide, Magnesium Chloride and Sodium Chloride, Diethyl Glycol, Dimethyl Acetamide, Dimethyl Sulfoxide (DMSO), Disaccharide, Erythritol, Ethanol, Ethylene Glycol, Formamide, Fructose, Glucose, Glycerol, Glycerol 3-phosphate,Glycol such as Diethyl Glycol or Triethylene Glycol, Glycine, Lactose, L-tyrosine, Lysine hydrochloride, Mannitol, MDP (2-Methyl-2,4-pentanediol), Phenylalanine, Planic acid, Polyethylene glycol such as PEG4000, polyethylene glycol succinate, folate-modified distearoylphosphatidyl ethanolamine polyethylene glycol, Polyethyleneimine (PEI), Polyvinylpyrrolidone (PVP), Proline, Propylene glycol, Pyridine (Pyridine-N-Oxide), Ribose, Sarcosine, Serine, Serum albumin, Sodium bromide, Sodium chloride, Sodium dodecylsulfonate, Sodium glutamate, Sodium iodide, Sodium sulfate, Sorbitol, Starch (hydroxyethyl starch), Sugar, Sucrose, Trehalose, Triethylene glycol, Trimethylamine, Tween 80 Tryptophan, Valine, and Xylose, and a combination or derivative of any of these compounds. A composition according to claim 1, characterized in that it is in the form of a powder or a liquid suspension. A composition according to any one of claim 1, wherein 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 water in the composition is greater than the volume occupied by at least one chain of said composition, and - the percentage by weight of water in the composition is greater than the percentage by mass of at least one chain of said composition.

4. Composition according to claim 1, wherein at least one chain is in powder form, and the composition has at least one of the following properties: - the volume occupied by water in the composition is less than the volume occupied by at least one chain of said composition, and - the mass percentage of water in the composition is less than the mass percentage of at least one chain of said composition, wherein the composition is preferably freeze-dried, desiccated, dried or dehydrated.

5. Composition according to any one of claims 1 to 4, wherein the nanoparticle core comprises a first free radical production or capture center Cifrpc, wherein Cifrpc is preferably selected 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.

6. Composition according to claim 5, wherein the coating comprises a second free radical production or capture center C2frpc •

7. Composition according to any one of claims 5 to 6, wherein Cifrpc and / or C2frpc is / are at least one antioxidant compound.

8. Composition according to any one of claims 5 to 7, wherein Cifrpc and / or C2Frpc is / are at least one oxidizing compound.

9. Composition according to any one of claims 1 to 8, wherein the nucleus is synthesized by a living organism or nanoparticle-producing cells, preferably a magnetotactic bacterium.

10. A composition according to any one of claims 5 to 9, wherein Cifrpc and / or C2Frpc is / are photosensitizer(s), selected from the group consisting of: 1) acridine, such as acridine orange, acridine yellow, 2) ALA (5-aminolevulinic acid), 3) aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) amino-olevulinic acid, delta-aminolevulinic acid, 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) Benzoporphyrin derivative with monoacid ring A, 18) Monoaspartyl chlorin(e6), 19) Talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrin, 23) Chlorine e6 porphrin, 24) Coal tar and derivatives, 25) Contraceptives, oral and estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview, 29) Dyes such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squarains, 33) BODIPY dyes, 34) Phenalenones, 35) Benzophenoxazinium dyes, 36) Erythrosine, 37) Flavines, 38) Foscan, 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(l-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) Chloroaluminum sulfonated phthalocyanine (CASP), 66) Silicon phthalocyanine (PC4), 67) RLP068, 68) Sodium porfimer, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-tetrakis(l-methylpyridinium-4-yl) porphyrin tosylate, 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, Rh(II)-Rh(II) cyclometal bridged dimeric compounds, platinum(II), gold(III), 87) Verteporfin, 88) Vulcan-based compounds such as aminovulinic acid, aminovulinic acid, 89) WST11, and 90) Xanthene.

11. A composition according to any one of claims 5 to 10, wherein CiFRPC and / or C2Frpc is / are a sound sensitizer(s), selected 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) non-steroidal anti-inflammatory drug, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) TiO2 Au-doped, 17) Carbon-based nanomaterial, 18) Carbon nanotube, 19) Chlorine, 20) Ce6, 21) PTX, Paclitaxel, 22) Chemotherapeutic drug or compound, 23) Infrared dye or IR783, 24) Curcumin, 25) Cyanine or Cu-Cyanine, 26) DHMS, 27) Dimethyl sulfide, 28) Docetaxel, 29) Chemotherapeutic drug or compound, 30) DOX / Mn-TPPS@RBCS, 31) Doxorubicin, 32) Manganese,33) globule, 34) red blood cell, 35) polymer, 36) elastomer, 37) erythosin or erythosin B, 38) FA or FA-OI or FA-OI NP or folic acid, 39) F3-PLGA@MB / Gd NPs, 40) poly(lactic-co-glycolic acid), 41) gadolinium, 42) Fe-TiO2 or titanium dioxide, 43) Fe-VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49) hydroxychloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysaffron 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) HSA-wrapped metal-porphyrin complex, 66) Albumin, 67) MnWOx, 68) MnWOx-PEG, 69) PEG, 70) Bimetallic oxide, 71) Mn(III)-HFs, 72) Manganese, Hemoporphine, 73) Nanobeads, 74) Noble metal nanomaterial, 75) OI NP or Oxygen indyocyanine, 76) Phta-locyanines,77) PIO or Pioglitazone, 78) Polymeric nanomaterial, 79) Porphyrin, 80) Pt2-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 isomer or Tris(l-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.

12. Composition according to any one of claims 5 to 11, wherein C1FRC and / or C2FRC is / are radiosensitizers, selected 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) Ga-dolynium, 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 doped lanthanide 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) Ni-motuzumab, 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) Membrane-active agent, 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) Sulphur group suppressors, 62) SYM004, 63) Texaphyrins, 64) TH-302, and 65) Tirapazamine.

13. A method for manufacturing the composition according to any one of claims 1 to 12, comprising at least one of the following steps: Step 1 of amplifying magnetotactic bacteria in at least one medium, comprising: the compounds necessary for the growth of magnetotactic bacteria and the production of magnetosomes, which are preferably selected 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 their combinations, preferably at a concentration between 1 nM and 2 Mol / L; - an iron source chosen from the group consisting of: at least one compound comprising at least one iron atom, iron citrate, iron quinate, iron chloride, iron sulfate, FeC13, and their combinations, at a concentration preferably between 1 nM and 2.10-3 Mol / L; - a nitrogen source chosen from the group consisting of: at least one compound comprising at least one nitrogen atom, a nitrate salt, nitrogen gas, ammonium, ammonia, an ammonium salt, urea, an amino acid, ammonia gas, and combinations thereof, preferably at a concentration between 1 nM and 4 Mol / L; -an oxygen source 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 some 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 potassium source 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 sulfur or sulfate source 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 manganese source 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 vitamin source 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 combinations, preferably at a concentration between 1 nM and 10-4 Mol / L, and -a calcium source consisting of at least one compound comprising at least one calcium atom, preferably at a concentration between 1 nM and 10-1 Mol / L. at least one compound necessary for doping magnetosomes with C ifrpc or another metal other than iron, preferably zinc or aluminum, for example a zinc source, preferably zinc sulfate or zinc citrate or zinc chlorate or zinc quinate. Step 2 of magnetosome extraction from magnetotactic bacteria and purification of the extracted magnetosomes by heating to obtain magnetosome minerals comprising a mass percentage of organic matter from magnetotactic bacteria of less than 1%, Step 3 involves coating magnetosome minerals with a coating material comprising compound C2frpc by mixing the magnetosome minerals with the coating material, wherein 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 amount or mass of the magnetosome minerals and the amount 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 freeze-dried composition obtained in step 5, preferably in water.

14. A method for storing the composition according to any one of claims 1 to 13, comprising at least one of the following steps: Step 1: Selecting or preparing the composition in the form of a liquid suspension, Step 2: Freeze-drying, desiccation, dehydration of the composition, or removal of water from the composition. Step 3: Store the composition in powder form, preferably for more than 3 months, and Step 4: Suspending or re-suspending the composition, preferably in water.