Particles comprising at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one compound for medical or cosmetic use

Abstract

The present invention relates to the use of particles as probes, in particular for in vitro use, said particles comprising at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one compound, wherein after a physicochemical perturbation applied to said particles, said compound dissociates from said iron oxide nanoparticle and / or is itself chemically modified.

Description

Technical Field

[0001] The field of the invention relates to particles comprising at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one compound and their medical or cosmetic uses. Background Art

[0002] The performance of some new probes relies on local detection of parameters such as temperature. To achieve this, fluorescent thermometers have been developed that contain iron oxide nanoparticles associated with fluorophores using methods that generally avoid quenching of luminescence by iron oxide (SK Mandal et al., Langmuir, Vol. 21, p. 4175 (2005)). However, these nanoparticles are superparamagnetic and therefore weakly magnetic. They have unstable magnetic moments at physiological or ambient temperatures.

[0003] A new mode of drug delivery has also been proposed by associating drugs with nanoparticles including magnetosomes to improve the efficacy of the drugs (JB. Sun et al., Cancer Letters, Vol. 258, p. 109 (2007)). However, in this case, the drugs cannot be released in a controlled manner. Summary of the invention

[0004] An object of the present invention is the use of a particle as a probe, in particular for in vitro use, said particle comprising:

[0005] At least one ferrimagnetic iron oxide nanoparticle associated with at least one compound, wherein said compound dissociates from said iron oxide nanoparticle and / or undergoes chemical modification itself upon application of a physicochemical perturbation to said particle.

[0006] More specifically, an object of the present invention is the use of a particle as a probe, in particular for in vitro use, said particle comprising:

[0007] At least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one compound, wherein said nanoparticle is preferably not bound to said compound via at least one nucleic acid or at least one amino acid, wherein upon application of a physicochemical disturbance to said particle, said compound dissociates from said iron oxide nanoparticle and / or undergoes chemical modification itself, wherein said dissociation and / or chemical modification of said compound does not preferentially cause its destruction, wherein said physicochemical disturbance preferentially induces a change in the state of said nanoparticle.

[0008] The granules according to the invention may contain at least 1, 10, 10 2 10 3 10 4 10 510 6 10 9 10 20 , or 10 40 ferrimagnetic or ferromagnetic iron oxide nanoparticles and / or at least 1, 10, 10 2 10 3 10 4 10 5 10 6 10 9 10 20 , or 10 40 A compound.

[0009] The particle according to the invention differs from the commonly described luminescent nanoparticle probe in its use when associated with a luminescent substance. In fact, its original mode of use relies on detecting changes in luminescence caused by the dissociation and / or chemical modification of the luminescent substance associated with the particle after applying a physicochemical perturbation to the particle.

[0010] In one embodiment, the luminescence and the change are less than 10 -10 %, 10 -5 %, 10 -1 %, 1%, 10%, 25%, 50%, 75%, or 90% of at least one luminescence characteristic, and the luminescence change is greater than 10 -10 %, 10 -5 %, 10 -1 %, 1%, 10%, 25%, 50%, 75%, or 90% of at least one luminescence characteristic of the particle, wherein the percentage change can be measured as a function of time or between at least two different spatial locations of the particle.

[0011] The particle according to the invention also differs from the nanoparticle associated with the drug in its use when it is associated with the drug compound. In fact, its original mode of use relies on the dissociation of the drug compound from the particle after applying a physicochemical perturbation to the particle.

[0012] The in vitro use of the particles may correspond to any use of the particles taking place outside a human being or an animal, without any contact with the human being or animal or any administration in the human or animal organism.

[0013] The in vitro use of the particles according to the invention may correspond to any use of the particles that occurs before the particles are administered or introduced into an organism or a part of the organism, or after the particles have exited from the organism or a part of the organism after the particles have been administered or introduced into the organism or a part of the organism. Preferably, the organism is a human or animal organism. Preferably, the part of the organism is a biological substance, DNA, RNA, protein, enzyme, lipid, tissue, cell, organelle, blood, or a tumor.

[0014] In one embodiment of the invention, the in vitro use of the particles may correspond to a medical treatment, such as a diagnosis or treatment, which is localized to a part of an organism and does not affect the entire organism or affects less than 90%, 50%, 25%, 10%, 5%, 1%, or 0.1% of the organism or the cells of the organism. The in vitro use of the particles may correspond to a medical treatment that affects at least one cell (preferably a prokaryotic cell or eukaryotic cell), tissue, organ or at least one biological substance, such as a cell, tissue, or cancerous organ or such as a bacterium or virus, different from the cells, tissues, organs or biological substances present in the organism in a normal state.

[0015] In one embodiment, the use of the particles according to the invention as probes makes it possible to detect, measure parameters such as temperature, pH value, chemical environment, or radiation. In some cases, it can then be used as a diagnostic tool.

[0016] In another embodiment, the use of the particles according to the invention as probes makes it possible to change parameters such as the state or physiological state of a cell. In some cases, it can then be used as a therapeutic tool.

[0017] The particles according to the present invention are aggregates of atoms and molecules in solid, liquid or gaseous state, preferably in solid state.

[0018] In one embodiment of the invention, the particles are not or do not comprise a crystalline liquid or a dendritic polymer.

[0019] The compound is a luminescent substance and / or a pharmaceutical compound.

[0020] The luminescent substance is a substance that is able to emit light when it is or has been exposed to light radiation. It can be phosphorescent or fluorescent.

[0021] The pharmaceutical compound is a compound for medical applications, in particular for diagnosis or therapy, in particular for use as a medical device or medicament, or for cosmetic or biological applications, in particular for research purposes.

[0022] In one embodiment of the present invention, NANOF refers to ferrimagnetic or ferromagnetic iron oxide nanoparticles, sometimes also equivalently referred to as nanoparticles, said nanoparticles or iron oxide nanoparticles.

[0023] The ferrimagnetic or ferromagnetic iron oxide nanoparticles according to the invention, represented herein by NANOF, have at least one of the following properties: (i) a generally crystalline core, wherein the crystallinity may be characterized by the presence of at least one crystal face, preferably at least five crystal faces, more preferably at least ten crystal faces, and its composition is iron oxide, preferably at least partially composed of maghemite, magnetite or a mixture of maghemite and magnetite, most preferably a composition consisting mainly of magnetite; (ii) a coating, sometimes also referred to as a membrane, which is of biological origin, composed of, for example, lipids and proteins; or of non-biological origin, wherein the coating may enable the transfer of energy or particles between the compound and the NANOF, stabilize the NANOF, render the NANOF biocompatible, promote the cellular internalization of the NANOF, and enable the coupling of the NANOF to different types of substances, including luminescent substances, substances of medical interest, substances with low natural abundance; (iii) a generally ferrimagnetic or ferromagnetic core. behavior, especially at physiological temperature; (iv) a single magnetic domain called a monodomain; (v) a possible chain arrangement that prevents or limits aggregation and may promote cellular internalization; (vi) a size measured in the presence or absence of a coating of 1 nm to 1 μm, 10 nm to 200 nm, 20 nm to 150 nm; (vii) there may be a charge, preferably a surface charge, wherein the charge may be a negative charge, especially when the nanoparticles are coated with negatively charged lipids, wherein the charge may vary with pH, ​​the presence or absence of a coating, and may promote cellular internalization, especially when it is positive or negative; (viii) at least one metal atom (such as an iron atom) and at least one other atom, such as an oxygen atom, whose magnetic moments are preferably antiparallel, thereby producing a property that may be ferrimagnetic; or (ix) at least one metal atom (such as an iron atom) and at least one other atom, such as another iron atom, whose magnetic moments are preferably parallel, thereby producing a property that may be ferromagnetic.

[0024] In one embodiment of the invention, the iron oxide nanoparticles are fully ferrimagnetic or fully ferromagnetic, but in some cases it may happen that the orientation of the magnetic moment is different between the surface and the core of the nanoparticle, for example when considering iron oxide nanoparticles whose core contains iron and whose surface is iron oxide.

[0025] In one embodiment of the present invention, the iron oxide nanoparticles are ferrimagnetic or ferromagnetic at a temperature below the Curie temperature or melting temperature of the nanoparticles or below the blocking temperature of the nanoparticles or at a pH value of 7, 6, 5, 4, 8, 9, or 10, or when the anisotropy energy of the nanoparticles is higher than the thermal energy of the nanoparticles, thereby generating a thermally stable magnetic moment of the nanoparticles.

[0026] In some embodiments of the invention, it may happen that the nanoparticle loses its ferromagnetism or ferrimagnetism, for example when it dissolves in vivo, when it is internalized into cells or lysosomes, in particular at pH values ​​that differ significantly from physiological pH values.

[0027] The ferrimagnetic or ferromagnetic behavior of NANOFs produces a thermally stable magnetic moment, especially at physiological temperature, and produces coercivity, residual magnetization, saturation magnetization, magnetocrystalline anisotropy, and the ability to generate heat under an applied alternating magnetic field, one or more of which are non-zero and preferentially high. The NANOF may thus have a coercive force greater than 1 Oe, 10 Oe, 100 Oe, 200 Oe, 500 Oe, 1000 Oe, or 10,000 Oe; a ratio between residual magnetization and saturation magnetization greater than 0.01, 0.1, 0.2, or 0.5; a saturation magnetization greater than 0.1 emu, 1 emu, 10 emu, 20 emu, 50 emu, 60 emu, 80 emu, 90 emu, or 100 emu per gram of NANOF; or a specific absorption rate (SAR) greater than 0.01 Watt, 0.1 Watt, 1 Watt, 10 Watt, 100 Watt, 200 Watt, 500 Watt, or 1000 Watt per gram of NANOF. The coercive force, the ratio between the residual magnetization and the saturation magnetization, or the saturation magnetization is preferably measured at a temperature above 0.1 K, 1 K, 10 K, 50 K, 100 K, 200 K, 400 K, or 800 K. The SAR is preferably measured at a NANOF concentration of greater than 0.01 mg, 0.1 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 50 mg, 100 mg, or 200 mg per milliliter, preferably by applying an alternating magnetic field having a strength above or below 0.1 mT, 1 mT, 2 mT, 5 mT, 10 mT, 20 mT, 50 mT, 100 mT, 1000 mT, or 10000 mT and / or a frequency above or below 1 Hz, 10 Hz, or 100 Hz, 1 KHz, 10 KHz, 100 KHz, 1000 KHz, or 10000 KHz. These magnetic properties are generally superior to those of superparamagnetic iron oxide nanoparticles known as SPIONs, which are commonly used for applications including medical applications. SPIONs are smaller in size than NANOFs and have magnetic moments that are generally thermally unstable at physiological temperatures.The ferrimagnetic or ferromagnetic behavior of NANOF can also produce at least one of the following properties: (i) interaction between the magnetic moment of NANOF and the dipole magnetic moment of the luminescent substance, which causes quenching of the luminescence of the luminescent substance, which is generally more effective than for SPION; (ii) coupling between the magnetic moment of NANOF and radiation (preferably a magnetic field, more preferably an alternating magnetic field), which makes it possible to induce luminescence changes, which is preferentially more important than for SPION; (iii) it is easier to induce the movement of the compound or the particle in the organism by applying a magnetic field, preferably a magnetic field gradient, more preferably an alternating magnetic field gradient; or (iv) promoting the cellular internalization of the compound or the particle by applying a magnetic field, preferably an alternating magnetic field, which can particularly enable the compound or the particle to measure changes in the environment of the compound or the particle or the characteristics of the radiation to which it is exposed within the cell.

[0028] Saying that the NANOF is associated with the compound is equivalent to saying that the compound is associated with the NANOF. In this case, the compound has at least one of the following properties. It can be: (i) bound to the NANOF; (ii) interacting with the NANOF; (iii) integrated into the NANOF, in particular during the preparation of the particles; (iv) integrated into the NANOF; (v) obtained from the NANOF, in particular when it is detached from the NANOF during the use of the particles; (vi) incorporated into the crystalline core of the NANOF or adsorbed onto the crystalline core of the NANOF; (vii) incorporated into the NANOF or adsorbed onto the coating covering the NANOF; (viii) in the environment of the NANOF; (ix) in direct contact with the NANOF, that is to say, in particular, without any substance between the compound and the NANOF; or (x) in indirect contact with the NANOF, that is to say, in particular, at least one substance, polymer or solvent is located between the NANOF and the compound.

[0029] In one embodiment of the invention, the substance, polymer or solvent between the NANOF and the compound is not synthesized by a living organism, is not a protein, a lipid, an enzyme, a DNA, a DNA strand, an RNA, an RNA strand, one or more nucleic acids, one or more amino acids, is not an active biological substance, or is not streptavidin or biotin. In this case, the NANOF may not be bound to the compound through at least one such substance, polymer or solvent between the NANOF and the compound.

[0030] In one embodiment of the present invention, NANOF is not formed by at least 1, 10, 10 2 10 3 10 610 10 10 20 , or 10 40 An amino acid, nucleic acid, DNA strand, RNA strand, protein, lipid, or enzyme is bound to the compound.

[0031] In one embodiment of the present invention, the substance, polymer, or solvent located between the NANOF and the compound can be at least one atom, such as carbon, or a complex of atoms produced by a living organism, protein, lipid, DNA, or RNA, wherein the atom or the complex of atoms has been denatured, preferably by chemical treatment and / or thermal treatment.

[0032] In one embodiment of the invention, the substance, polymer or solvent located between the NANOF and the compound represents less than 90%, 50%, 25%, 10%, 5%, 1%, or 0.1% of the mass of NANOF, wherein the percentage may correspond to the mass of the substance, polymer, or solvent divided by the mass of NANOF.

[0033] In one embodiment of the invention, the substance, polymer, or solvent between the NANOF and the compound is not denatured or destroyed after the physicochemical disturbance is applied to the particle, or is not the cause of the dissociation of the compound from the NANOF and / or the chemical modification of the compound after the physicochemical disturbance is applied to the particle. The denaturation or destruction may be associated with the loss or reduction of activity of the substance, polymer, or solvent, such as the loss or reduction of activity of a protein, lipid, enzyme, conversion of double-stranded DNA to single-stranded DNA, or the loss of at least one amino acid or at least one nucleic acid from a complex of amino acids or nucleic acids.

[0034] In one embodiment, the NANOF is responsible for the dissociation of the compound from the NANOF and / or the chemical modification of the compound following application of a physicochemical perturbation to the particle.

[0035] In one embodiment of the present invention, when the distance between the compound and the NANOF is less than 10 μm, 1 μm, 100 nm, 10 nm, or 1 nm, the compound is in direct contact with the NANOF.

[0036] The environment of NANOF can be a liquid medium, a solid medium or a gas medium or at least one, ten, ten different from the compound. 2 10 3 10 6 10 10 Species, or 10 40A substance, preferably one or more substances, which surrounds or includes the particle at a distance preferably less than 1 m, 1 dm, 1 cm, 1 mm, 1 μm, 100 nm, or 10 nm measured from the center or outer surface of the particle.

[0037] The environment of NANOF can be a liquid medium, a solid medium or a gas medium or at least one, ten, ten different from the compound. 2 10 3 10 6 10 10 Species, or 10 40 A substance, preferably one or more substances, which surrounds or includes the particle at a distance preferably greater than 1 m, 1 dm, 1 cm, 1 mm, 1 μm, 100 nm, or 10 nm measured from the center or outer surface of the particle.

[0038] The environment of NANOF can be a liquid medium, a solid medium or a gas medium or at least one, ten, ten different from the compound. 2 10 3 10 6 10 10 Species, or 10 40 A substance, preferably one or more substances, which surrounds or includes the particle at a distance preferably less than or greater than the diameter of the earth, or 1000 km, or 1 km, or 1 m, or 1 dm, or 1 cm, or 1 mm, or 1 μm, or 100 nm, or 10 nm measured from the center or outer surface of the particle.

[0039] According to the present invention, a compound is considered to be dissociated from NANOF when the compound is separated from NANOF to end up in the environment of NANOF.

[0040] In one embodiment of the invention, the particles are inactivated or in an inactivated state when no physicochemical disturbance is applied to the particles.

[0041] In another embodiment, the particles are activated or in an activated state upon application of a physicochemical perturbation to the particles.

[0042] The probe according to the invention may contain particles in an activated state and / or an inactivated state.

[0043] According to the present invention, the modified or unmodified NANOF and / or compound and / or magnetosome can be in an activated state or an inactivated state.

[0044] Physicochemical perturbations can be imposed using artificial means, such as MRI, or naturally by the natural environment of the particle, such as a radioactive environment.

[0045] The dissociation of the compound from NANOF can be determined by the dissociation constant K D To characterize the dissociation constant K D It can be equal to the ratio of the concentration of the compound bound to NANOF measured when the particle is activated divided by the same concentration measured when the particle is inactivated. Preferably, the constant K according to the present invention is D Can be lower than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.02, 0.1, 0.05, 0.01, 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , or 10 -9 According to this definition, K D The smaller it is, the greater the dissociation of the compound from NANOF.

[0046] A measure of the bond or interaction strength between the compound and NANOF can be given by the dissociation constant K D The value of K is given. D The larger the value, the stronger the binding strength between the compound and NANOF.

[0047] A compound is considered to be chemically modified itself when the composition and / or structure of the compound is changed, thereby inducing a change in some of its properties, in particular the luminescent properties and / or the pharmacological properties.

[0048] In one embodiment of the invention, the dissociation and / or chemical modification of the compound does not cause its destruction or denaturation or retains the compound. In this case, the dissociation and / or chemical modification of the compound does not preferentially cause its denaturation or destruction of all or part of it, or the loss of at least 1, 10, 10 2 10 5 10 10 , or 10 20 The physicochemical disturbance is preferably a disturbance caused by a change in the environment of the particle, the NANOF or the compound.

[0049] The physicochemical disturbance applied to the iron oxide NANOF can preferentially and selectively induce a change in the state of the NANOF without inducing such a change in state in the compound. This may be the case when the NANOF is exposed to a physicochemical disturbance that induces a change in the state of the NANOF, such as movement or temperature increase. The physicochemical disturbance can induce a greater change in state in the NANOF than in the compound, such as a greater temperature change, a more important movement, or a more important change in characteristics. In order to confirm this difference in state change, a physicochemical disturbance can be applied to a separate NANOF and a separate compound and a change in one of the characteristics of the separate NANOF can be measured after applying the disturbance and the change is more important than the change in the same characteristic of the separate compound.

[0050] The physicochemical interference is preferably an interference that does not irreversibly change the properties of the compound or NANOF, preferably the luminescent properties or therapeutic properties of the compound. For example, such interference that causes the denaturation of the compound or NANOF is preferably avoided, the interference being to increase the temperature to a melting temperature or denaturation temperature that exceeds the compound or NANOF. Preferably, the physicochemical interference is moderate enough so that the compound at least partially maintains its properties, preferably the properties of a luminescent or therapeutic substance, after applying the interference.

[0051] The physicochemical disturbance is preferably a sufficiently mild disturbance, such as a moderate increase in the temperature of the particles, preferably below 300°C, 200°C, 100°C, 50°C, 20°C, 10°C, or 1°C, which may cause a mass loss of less than 90%, 50%, 25%, 10%, 5%, 1% or 0.1% of the particles.

[0052] The radiation may be wave radiation or particle radiation, sound wave radiation, ionizing radiation, X-ray radiation, electromagnetic radiation, radiation due to electric, magnetic or electromagnetic fields, or radiation caused by neutrons, protons, electrons, positrons, alpha particles, beta, gamma, neutrinos or muons.

[0053] The environment of the particle may be a liquid medium, a solid medium or a gaseous medium or at least one, 10, 10 or more of the following surrounding or including the particle at a distance measured from the center or outer surface of the particle, preferably less than: 2 10 3 10 4 10 5 10 6 10 9 10 20 Species, or 10 40types of substances: 10 000km, 1000km, 100km, 10km, 1km, 1m, 1dm, 1cm, 1mm, 1μm, 100nm, or 10nm.

[0054] The environment of a particle according to the invention may be a liquid medium, a solid medium or a gaseous medium or at least one, ten, ten or more of the following surrounding or including said particle at a distance measured from the center or the outer surface of the particle, preferably greater than: 2 10 3 10 4 10 5 10 6 10 9 10 20 Species, or 10 40 types of substances: 10 000km, 1 000km, 100km, 10km, 1km, 1m, 1dm, 1cm, 1mm, 1μm, 100nm, or 10nm.

[0055] The environment of a particle according to the invention may be a liquid medium, a solid medium or a gaseous medium or at least one, ten, ten or more of the following surrounding or including the particle at a distance measured from the center or the outer surface of the particle that is preferably greater or less than: 2 10 3 10 4 10 5 10 6 10 9 10 20 Species, or 10 40 Types of material: diameter of the earth, or 10000km, or 100km, or 10km, or 1km, or 1m, or 1dm, or 1cm, or 1mm, or 1μm, or 100nm, or 10nm.

[0056] The environment of the compound according to the invention may be a liquid medium, a solid medium or a gaseous medium or at least one, ten, ten or more of the following surrounding or including the compound at a distance measured from the center or the outer surface of the compound, preferably greater or less than: 2 10 3 10 4 10 5 10 6 10 9 10 20 Species, or 10 40types of material: diameter of the earth, or 10 000km, or 1000km, or 10km, or 1km, or 1m, or 1dm, or 1cm, or 1mm, or 1μm, or 100nm, or 10nm.

[0057] The environment of the particle according to the invention may comprise said particle, said compound and / or NANOF.

[0058] The substances of the environment of the particle, compound or NANOF according to the invention can be atoms, molecules, polymers contained in the environment; chemical substances or biological substances, preferably substances different from the compound or NANOF; DNA, RNA, proteins, lipids, enzymes, or nucleic acids or amino acids.

[0059] The environment of the particles, compounds or NANOFs according to the invention may be a biological environment, that is to say an environment comprising at least one biological substance, such as cells, organelles, DNA, RNA, proteins, lipids, amino acids, or nucleic acids.

[0060] The change in the environment of the particles, NANOF or compounds according to the present invention can be a pH change, preferably a change of less than or greater than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.1 pH units; a temperature change, preferably a change of less than or greater than 500°C, 400°C, 300°C, 200°C, 100°C, 50°C, 25°C, 10°C, 5°C, 1°C, or 0.1°C; a change in redox potential, preferably a change of less than or greater than 1000V, 100V, 10V, 5V, 2V, 1V, 0.1V, or 0.01V; a change in viscosity, preferably a change in dynamic viscosity, preferably less than or greater than 10 20 Pa.s, 10 10 Pa.s, 10 5 Pa.s, 10 3 Pa.s, 10 2 Pa.s, 10Pa.s, 10 -1 Pa.s, 10 -2 Pa.s, 10 -3 Pa.s, 10 - 6 Pa.s, 10 -9 Pa.s, or 10 -20 The change in the chemical composition of the environment may correspond to a change in the concentration of at least one substance in the environment and may preferably be less than or greater than 100M, 10M, 1M, 10 -1 M, 10 -2M, 10 -3 M, 10 -6 M, or 10 -9 M.

[0061] In this specification we specifically distinguish between the situation wherein the particles are inactivated and have not undergone any physicochemical perturbation and the situation wherein the particles are activated and have undergone such a perturbation.

[0062] The conditions under which the particles according to the invention function correspond to the conditions under which said particles are transformed from an inactivated state into an activated state or vice versa from an activated state into an inactivated state.

[0063] For certain modes of use of the particles according to the invention, the compound has a stronger interaction with the NANOF when the particles are inactivated than when the particles are activated. This may be the case in particular when the use of the particles involves the dissociation of the compound from the NANOF.

[0064] For other modes of use of the particles according to the invention, the interaction between NANOF and the compound may not change or may even increase when the particles are converted from an inactive state to an activated state. This may be the case in particular when the use of the particles involves chemical modification of the compound.

[0065] The schematic diagram is shown in Figure 1 , which illustrates the use of the particles according to the invention involving the dissociation of the compound from NANOF or the chemical modification of the compound.

[0066] In one embodiment of the present invention, the ferrimagnetic or ferromagnetic iron oxide nanoparticles (NANOF) are synthesized by magnetotactic bacteria and do not contain any carbonaceous material derived from said bacteria.

[0067] In one embodiment of the present invention, the NANOF is synthesized by a living organism, preferably a bacterium, more preferably a magnetotactic bacterium.

[0068] According to one embodiment of the present invention, NANOF is a nanoparticle synthesized by magnetotactic bacteria, which is particularly selected from the group consisting of: Magnetospirillum magneticum AMB-1, coccus MC-1, vibrios MV-1, MV-2 and MV-4, Magnetospirillum MS-1, Magnetospirillum gryphiswaldense MSR-1, spirillum, Magnetospirillum MGT-1 and Desulfovibrio magneticus RS-1.

[0069] The NANOFs synthesized by magnetotactic bacteria are magnetosomes, in particular as defined by DA Bazylinski et al., Nature Reviews Microbiology, Vol. 2, p. 217 (2004).

[0070] The NANOF according to the present invention can be chemically synthesized or synthesized by another living organism other than magnetotactic bacteria and has at least one property that is the same or similar to that of modified or unmodified magnetosomes.

[0071] The magnetic body according to the present invention may have at least one property that is the same as or similar to that of NANOF.

[0072] In one embodiment of the present invention, the magnetosomes are in the form of chains comprising 2 to 1000 magnetosomes, 2 to 500 magnetosomes, 2 to 250 magnetosomes, 2 to 100 magnetosomes, 2 to 75 magnetosomes, typically 4 to 20 magnetosomes, which are preferentially maintained in chains by materials of biological origin (such as lipids and proteins) or non-biological origin.

[0073] The magnetosomes belonging to these chains usually have a crystallographic direction, an easy magnetization axis, or a field line oriented in the direction of chain elongation. This orientation makes it possible, among other things, to stabilize the magnetic moment of the magnetosomes. Thus, the magnetosome chains have a pronounced magnetic anisotropy. When several magnetosome chains containing at least X magnetosomes interact, this leads to the formation of longer magnetosome chains containing at least X+1 magnetosomes.

[0074] The length of the magnetosome chain according to the present invention can be less than or greater than 1m, 1dm, 1cm, 1mm, 100μm, 10μm, 1μm, 600nm, 300nm. Due to their chain arrangement, the magnetosomes do not aggregate and have a stable magnetic moment that can couple with the magnetic field itself, thereby promoting the luminescence change of the probe. The chain arrangement also promotes the cellular internalization of the magnetosomes, which is enhanced by applying an alternating magnetic field.

[0075] In one embodiment of the present invention, in order to determine whether the NANOF or magnetosome is in the form of a chain, a few microliters of the NANOF suspension is deposited on a carrier (usually a carbonaceous net) and the NANOF is analyzed under an electron microscope. When the NANOF has a crystallographic axis or direction oriented in the direction of chain elongation, in particular at least 1, 2, 5, 10, 20, 30, 50 directions, the NANOF is considered to be arranged in a chain. Such an orientation may mean that the angle between the crystallographic direction of the NANOF and the chain elongation direction is less than 90 °, 80 °, 50 °, 40 °, 20 °, 10 °, or 5 °. NANOF without this characteristic is not arranged in a chain.

[0076] In one embodiment of the invention, the magnetotactic bacteria may be cultured in a culture medium, for example as described in patent application WO 2011 / 061259, which is incorporated herein by reference.

[0077] In one embodiment of the invention, the culture medium of the magnetotactic bacteria contains at least one iron source, such as a solution of ferric quinate, ferric citrate, ferric chloride, ferric malate, or ferrous sulfate, in particular a solution at a concentration of 0.02 μM to 2 μM, 0.02 μM to 200 mM, 0.2 μM to 20 mM, 2 μM to 2 mM, or 20 μM to 200 μM, and one or more other additives, usually added at a concentration of 0.01 μM to 1 M, 0.01 μM to 100 mM, 0.01 μM to 10 mM, 0.01 μM to 1 μM, 0.1 μM to 100 μM, or 1 μM to 10 μM. These additives are preferably selected from transition metals, chelating agents, luminescent substances, substances / isotopes of low frequency / natural abundance, substances having the characteristics of several of the above substances. The bacteria or these additives can be introduced into the culture medium during at least one of the different growth phases of the magnetotactic bacteria, such as the latent phase, the acceleration phase, the exponential phase, the decay phase or the stationary phase and / or the bacteria can be cultured in the presence of a low concentration of oxygen below 1000 bar, 100 bar, 10 bar, 1 bar, 200 mbar or 20 mbar and / or the pH value or another parameter of the growth medium of the magnetotactic bacteria is maintained at a constant or almost constant value during the growth of the magnetotactic bacteria, for example by adding an acidic solution or an alkaline solution, which solution preferably contains a certain concentration of these additives. Furthermore, these additives and bacterial growth conditions are preferably chosen from those that make it possible to avoid toxicity, confer good magnetic and / or luminescent properties to the probe, allow the incorporation of the additives into the magnetotactic bacteria and / or magnetosomes, cause an increase in the size and / or length of the magnetosome chains, stimulate the growth of magnetotactic bacteria, make it possible to achieve a sufficiently high yield of magnetosomes, in particular a yield of more than 0.1 mg, 1 mg, 100 mg, 1 g, 10 g of magnetosomes per liter of culture, make it possible to obtain a narrow magnetosome size distribution, in particular below 60 nm, 30 nm, 15 nm, 10 nm, 5 nm, or 1 nm.

[0078] In one embodiment of the present invention, the association of the luminescent substance with the magnetosomes may or may not be obtained by genetic manipulation of the magnetotactic bacteria.

[0079] In one embodiment of the present invention, the luminescent substance is preferably not a fluorescent protein, such as green fluorescent protein or yellow fluorescent protein, a luciferase, a fluorescent amino acid, a fluorescent nucleic acid, a DNA or a fluorescent DNA chain, or an RNA or a fluorescent RNA chain.

[0080] Another object of the invention is a method for producing said particles, comprising culturing magnetotactic bacteria in the presence of a luminescent substance, followed by extracting said magnetosomes from these bacteria, purifying them and characterizing them.

[0081] Another object of the invention is a method for producing said particles, comprising the cultivation of magnetotactic bacteria in the absence of luminescent substances, followed by the extraction of magnetosomes from these bacteria, their purification, the association of luminescent substances with the magnetosomes and their characterization.

[0082] In one embodiment of the present invention, the NANOF is a modified magnetosome.

[0083] The modified magnetosomes are preferably magnetosomes which contain no, essentially no or little organic or carbonaceous material, in particular consisting of lipids, proteins, DNA, RNA or enzymes, which may in particular surround the mineral core of the magnetosomes.

[0084] In one embodiment of the invention, the mineral contains no carbon or contains less than 75%, 50%, 25%, 10%, 5%, 2%, 1%, or 0.5% carbon.

[0085] In one embodiment of the present invention, the modified magnetosomes according to the present invention contain less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1%, or 0.1% carbonaceous material, wherein the percentage of the carbonaceous material can be defined as the ratio between the number of carbon atoms contained in the modified magnetosomes and the total number of atoms contained in the modified magnetosomes.

[0086] The modified magnetosomes are preferably nanocrystals consisting mainly of a mineral core and possibly a coating, said coating not originating from said magnetotactic bacteria.The modified magnetosomes may have at least one property in common with the unmodified magnetosomes, such as chain arrangement.

[0087] The preparation of modified magnetosomes according to the present invention is preferably pyrogen-free and contains no, essentially no, or only small amounts of endotoxins or lipopolysaccharides.

[0088] In one embodiment of the present invention, the preparation of modified magnetosomes comprises less than 10 mg / ml, mg / mg, or cm3 of modified magnetosomes or the preparation of modified magnetosomes comprises less than 10 mg / ml, mg / mg, or cm3 of modified magnetosomes. 6 , 10 5 , 10 4 , 10 3 , 10 2 , 10, 1, 10 -1 , 10-2 , 10 -3 , 10 -4 , or 10 -5 EU (endotoxin unit).

[0089] In one embodiment of the present invention, the maximum dimension of the particles is preferably comprised between 1 nm and 10 μm or between 10 nm and 1 μm.

[0090] In one embodiment of the invention, the largest dimension of the particles is preferably less than 100 μm, 50 μm, 10 μm, 1 μm, 500 nm, 250 nm, 100 nm or 50 nm.

[0091] In one embodiment, the particles according to the invention further comprise a substance selected from the group consisting of low frequency or natural abundance isotopes, transition metals, and substances of medical interest.

[0092] Advantageously, a low frequency or naturally abundant isotope may be associated with the particle to facilitate detection of the particle in a medium containing little or no such isotope (eg, iron-54, iron-57, or iron-59, which occur minimally in nature).

[0093] Advantageously, transition metals may be associated with the particle to modify its magnetic properties, and in particular to facilitate detection of changes in its luminescence.

[0094] It is also advantageous that substances of medical interest, such as targeting agents, substances used during medical procedures, treatments, diagnostics, surgical procedures, radiation therapy, hyperthermia, MRI, chemotherapy, can be associated with the particles or compounds to particularly facilitate or improve its use or medical efficacy.

[0095] In one embodiment of the invention, the particles according to the invention may be encapsulated in a compartment, preferably a vesicle, most preferably a lipid vesicle, such as a liposome, a macrovesicle or a nanogel, in order in particular to increase the efficacy of the particle or to reduce the toxicity of the particle.

[0096] In one embodiment of the invention, the particles according to the invention associate, bind or interact with a substance that does not belong to the particle. This may make it possible to detect the substance or the state of the substance. This may also make it possible to improve the medical, therapeutic, diagnostic, magnetic, luminescent and heating properties of the particle, or to reduce the risk of toxicity, in particular by preventing direct contact between the particle and the organism.

[0097] In one embodiment of the present invention, the ferrimagnetic or ferromagnetic iron oxide nanoparticles (NANOFs) are chemically synthesized.

[0098] In one embodiment of the invention, the compound is associated with the NANOF via a weak bond. This situation is preferably encountered when the use of the particles according to the invention involves the dissociation of the compound from the NANOF and / or the chemical modification of the compound of the NANOF.

[0099] A weak bond is preferably a bond that allows the compound to dissociate from the particle under a physicochemical disturbance, wherein the disturbance is preferentially such that it does not significantly change or it changes in an insignificant way at least one of the properties of the NANOF, such as its size, size distribution, ferrimagnetism or ferromagnetism, composition, or crystal structure.

[0100] In one embodiment of the present invention, a significant change in the properties of NANOF can be defined as: (i) an increase or decrease in the size of the NANOF by more than 100 nm, 1 μm, or 10 μm; (ii) an increase or decrease in the size distribution of the NANOF by more than 100 nm, 1 μm, or 10 μm; (iii) a change in magnetic properties due to, for example, the transformation of the NANOF from a single-domain magnetic nanoparticle to a multi-domain magnetic nanoparticle, which can particularly cause a loss of ferrimagnetism or ferromagnetism of the NANOF; (iv) a change in composition, such as a transformation from nanoparticles composed of iron oxide to nanoparticles composed only of iron or oxygen; or (v) a change in crystal structure, such as a transformation from nanoparticles in a crystalline state to nanoparticles in an amorphous state, wherein the crystalline state can be characterized by the presence of at least two crystal planes in the nanoparticles and the amorphous state can be characterized by the absence of crystal planes in the nanoparticles.

[0101] In one embodiment of the present invention, insignificant changes in the properties of NANOF can be defined as: (i) the size of the NANOF increases or decreases by less than 100 nm, 1 μm, or 10 μm; (ii) the size distribution of the NANOF increases or decreases by less than 100 nm, 1 μm, or 10 μm; (iii) there is no change in magnetic properties due to, for example, the transformation of the NANOF from single-domain nanoparticles to multi-domain nanoparticles, which transformation can significantly cause the loss of ferrimagnetic or ferromagnetic properties of the NANOF; (iv) there is no composition change, such as a transformation from an iron oxide composition to an iron or oxygen composition; or (v) there is no change in crystal structure, such as the absence of a transformation from a crystalline state of nanoparticles to an amorphous state of nanoparticles, wherein the crystalline state can be characterized by the presence of at least two crystal planes in the nanoparticles, and the amorphous state can be characterized by the absence of crystal planes in the nanoparticles.

[0102] Preferably, the weak bond according to the present invention is a hydrogen bond, a bond due to Van der Waals force, London force, Debye force, intermolecular force or inter-nanoparticle force, or hydrophobic, hydrophilic, or electrostatic interaction.

[0103] In one embodiment of the present invention, the weak bond may be associated with the adsorption of the compound onto the NANOF.

[0104] In one embodiment of the present invention, the weak bond can be represented by the dissociation constant K D To characterize, the dissociation constant K D Preferentially less than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.02, 0.1, 0.05, 0.01, 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , or 10 -9 .

[0105] In one embodiment of the invention, the bond between the compound and the NANOF is stronger when the particle is deactivated than when the particle is activated. This property can be particularly observed when the use of the particle involves the dissociation of the compound from the NANOF.

[0106] In one embodiment of the invention, the compound is not bound to NANOF by a strong bond, such as a covalent bond, an ionic bond, or a bond that prevents the compound from dissociating from NANOF. This property is preferentially observed when the use of the particles involves dissociation of the compound from NANOF and / or chemical modification of the compound.

[0107] In one embodiment of the invention, the compound is associated with the NANOF via a strong bond. This is preferentially satisfied when the use of the particles involves chemical modification of the compound.

[0108] A strong bond is preferably one that prevents dissociation of the compound from the NANOF under a physicochemical perturbation, wherein the perturbation is preferentially one that does not significantly alter at least one of the properties of the NANOF, such as its size, size distribution, ferrimagnetism or ferromagnetism, composition, or crystal structure.

[0109] In one embodiment, the strong bond is a bond that is not a weak bond.

[0110] Preferably, the strong bond according to the present invention is a covalent bond, a metallic bond or an ionic bond.

[0111] In one embodiment of the invention, the strong bond can be represented by the dissociation constant K D To characterize, the dissociation constant K D Preferentially greater than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.02, 0.1, 0.05, 0.01, 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , or 10 -9 .

[0112] In one embodiment of the invention, the particles comprise a luminescent compound.

[0113] The luminescent substance according to the invention has at least one luminescent property.

[0114] A luminescence property is defined herein as intensity, luminescence wavelength, intensity, absorption wavelength, luminescence lifetime, or luminescence anisotropy, which is preferentially measurable. The luminescence property may be attributed to a luminescent substance or particle.

[0115] In one embodiment of the invention, the luminescent substance has at least one luminescence property which preferably varies under the operating conditions of the particle.

[0116] A luminescence change is defined herein as a change in at least one luminescence property. It may be attributable to a luminescent substance or particle and may be an increase or decrease, preferably an increase, of at least one luminescence property of the particle or luminescent substance.

[0117] The luminescence properties may vary over time, depending on the spatial position of the particle or NANOF, during changes in the particle or NANOF environment, following application of radiation, dissociation of the luminescent species from the NANOF and / or chemical modification of the luminescent species.

[0118] In one embodiment of the invention, the luminescence change is a continuous increase in luminescence. It preferentially occurs over time over a period of time that is preferably longer than: 10 -10 sec, 10 -9 sec, 10 -8 sec, 10 -7 sec, 10 -6 sec, 10 -5 sec, 10 -4 sec, 10 -3 sec, 10-2 sec, 10 -1 The present invention relates to a method for producing a light-emitting substance which is present in an amount of at least 1, 10, 20, 30, or 45 seconds, 1, 2, 5, 10, 30, or 45 minutes ... 2 , 10 3 , 10 6 , 10 9 , 10 20 , or 10 40 The luminescence increases and decreases.

[0119] In one embodiment of the invention, the change in luminescence over time (ΔL) is characterized by the slope ΔL / δt at the starting point, where δt represents the duration originating from the start of the luminescence change, which is preferably determined during the first few seconds or minutes after applying a physicochemical disturbance to the particles.

[0120] The luminescence property or its change can be determined around the particle or NANOF, at the surface of the particle or NANOF or in the particle or NANOF, at a distance preferably less than 1m, 1dm, 1cm, 1mm, 1μm, 100nm, or 10nm measured from the center of the particle or NANOF.

[0121] In one embodiment of the invention, the luminescence change is determined in an area that enables the detection of the luminescence change, i.e., an area that contains a sufficient number of particles, preferably greater than 1, 10, 100, 10 3 10 4 10 5 10 6 10 7 10 8 10 9 10 10 , or 10 11 particles or particles / unit volume, especially particles / m 3 , particles / cm 3 , particles / mm 3 , or particles / μm 3 .

[0122] In one embodiment of the present invention, the luminescent material has at least one luminescent property that is measurable over time and / or as a function of the position of the particle or NANOF; or undergoes a change in at least one of its properties that is measurable over time and / or as a function of the position of the particle or NANOF.

[0123] In another embodiment of the invention, certain parameters, such as the measurement time, the size and mass of the luminescent substance and the NANOF, the type of bond between the luminescent substance and the NANOF, or the characteristics of the environment of the particle, such as its viscosity, are optimized to allow measuring the luminescence changes of the luminescent substance.

[0124] In one embodiment of the present invention, the luminescent substance has at least one of the following properties: (i) a small size or a small mass, preferably less than 1 / 10, 1 / 100, or 1 / 1000 of that of NANOF, which can make it particularly possible to promote the dissociation of the luminescent substance from NANOF and / or its diffusion in the environment of the particle; (ii) a negative charge, preferably less than -1mV, -10mV, -20mV, or -50mV; a positive charge, preferably greater than 1mV, 10mV, 20mV, or 50mV; a neutral charge, or a charge opposite to that of NANOF, which can be particularly conducive to the bonding between the luminescent substance and NANOF; (iii) a bond between the luminescent substance and NANOF that enables it to dissociate from NANOF under the operating conditions of the particle; (iv) crystallinity revealed by the presence of at least two crystal planes; or (v) non-zero absorption at a wavelength where biological media absorb little, or non-zero absorption at 20nm to 2μm or 600nm to 1200nm.

[0125] In one embodiment of the present invention, a preferred luminescent material is one that achieves the strongest luminescence intensity quenching and / or the smallest luminescence lifetime when the particle is deactivated.

[0126] In one embodiment of the present invention, preferred luminescent materials are those that enable the most significant energy or particle transfer with the NANOF when the particle is deactivated or activated.

[0127] In one embodiment of the present invention, preferred luminescent materials are those that make it possible to induce a luminescence change that is: (i) as large as possible and / or measurable when the particle is transformed from an inactivated state to an activated state; or (ii) measurable when the particle is exposed to low intensity, low energy radiation and / or low amplitude changes in the particle's environment, such as changes of less than 10 6 ℃, 10 5 ℃, 10 4 ℃, 10 3 ℃, 10 2℃, 10℃, 5℃, 1℃, 0.5℃, or 0.1℃ temperature change or less than 100, 50, 20, 14, 10, 8, 6, 4, 2, 1, 0.5, 0.1, or 0.01 pH units.

[0128] In one embodiment of the invention, at least one luminescent property of the luminescent material associated with the iron oxide nanoparticles is different from the luminescent property of the luminescent material alone. This difference is more easily observable and measurable when the particles are inactivated or activated, preferentially inactivated, than when the particles are transformed from an inactivated state to an activated state or from an activated state to an inactivated state.

[0129] In one embodiment of the invention, the luminescence intensity of the luminescent substance associated with the iron oxide nanoparticles is quenched by the iron oxide nanoparticles compared to the luminescence intensity of the luminescent substance alone. This quenching is preferably observed and measurable when the particles are deactivated.

[0130] In one embodiment of the present invention, the quenching of the luminescence intensity of the luminescent substance is determined by measuring the luminescence intensity I of the luminescent substance bound to the NANOF. L The luminescence intensity I of the single luminescent substance not combined with NANOF is NL The ratio between L / I NL This ratio, especially I L Preferably it is measured when the particles are inactivated.

[0131] In one embodiment of the present invention, I L and I NL The measurements are made under identical conditions, in particular with regard to the parameters for the excitation and detection of the luminescence and the concentration of the luminescent substance used during the measurement.

[0132] In one embodiment of the present invention, when I L / I NL Less than 1, preferably less than 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.001, 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , or 10 -9 When , the luminescence intensity of the luminescent substance associated with NANOF is quenched.

[0133] In one embodiment of the present invention, the luminescence lifetime of the luminescent substance associated with the iron oxide nanoparticles is changed, quenched, or extended, preferably extended, by the iron oxide nanoparticles compared to the luminescence lifetime of the luminescent substance alone. In the case where the luminescence lifetime is extended by the iron oxide nanoparticles, this is an unusual situation, unlike the usual Förster transfer ( transfer) that causes a shortened lifespan.

[0134] In one embodiment of the present invention, the change in the luminescence lifetime of the luminescent substance is measured by measuring the luminescence lifetime T of the luminescent substance combined with NANOF. A The luminescence lifetime T of the single luminescent substance not combined with NANOF is L The ratio T A / T L To be determined.

[0135] In one embodiment of the present invention, T A and T L The measurements are made under identical conditions, in particular with regard to the parameters for the excitation and detection of the luminescence and the concentration of the luminescent substance used during the measurement.

[0136] In one embodiment of the present invention, when T A / T L Less than 1, preferably less than 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.001, 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , or 10 -9 When NANOF is used as a light emitting material, the luminescence lifetime of the luminescent material combined with NANOF is shortened, especially due to the presence of NANOF.

[0137] In one embodiment of the present invention, when T A / T L Greater than 1, preferably greater than 2, 4, 8, 10, 25, 50, 100, 1000, 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , or 10 9 When NANOF is used as a light emitting material, the luminescence lifetime of the light emitting material combined with NANOF is prolonged, especially due to the presence of NANOF.

[0138] In one embodiment of the present invention, at least one luminescent property of the luminescent material associated with NANOF changes compared to the luminescent material alone when the luminescent material has at least one of the following properties: i) transferring energy or particles to or from NANOF, and the energy or particle exchange may be a Foster transfer or a Dexter transfer; (ii) the separation distance between the luminescent material and NANOF is preferably less than 1 cm, 1 μm, 500 nm, 100 nm, 50 nm, 25 nm, 10 nm, or 5 nm, or the separation distance is preferably less than 100 times, 100 times, 50 times, 20 times, 10 times, 2 times the Foster distance; , or 1 times, wherein the Forster distance is preferably 0.1nm to 50nm, 0.5nm to 20nm, or 1nm to 10nm; (iii) the characteristics of the donor or acceptor in the terms used to describe Forster transfer; (iv) the ability to emit light at one or more wavelengths of light absorbed by the NANOF; (v) the lifetime or luminescence intensity measured when the luminescent substance is alone, which is sufficiently different from the lifetime or luminescence intensity of the luminescent substance combined with the NANOF so that the change in the lifetime or luminescence intensity is measurable; or (vi) the lifetime of the luminescent substance alone is greater than 0.5ns, 1ns, 5ns, 10ns, 100ns, 1μs, 10μs, 100μs, or 1ms.

[0139] In one embodiment of the present invention, at least one luminescent property of the luminescent material associated with the NANOF changes compared to the luminescent material alone when the NANOF has at least one of the following properties: (i) non-zero, preferentially significant absorption, absorption coefficient, absorption index, or extinction coefficient at the emission wavelength of the luminescent material, more preferably significant at 10 nm to 5000 nm, 20 nm to 2000 nm, or 100 nm to 1500 nm; (ii) containing materials in its core and / or coating that absorb at one or more emission wavelengths of the luminescent material; (iii) having a size greater than SPION, greater than 0 .1nm, 1nm, preferably greater than 10nm, more preferably greater than 20nm, which can make it possible in particular to increase its absorption, its absorption coefficient, its absorption index, or its extinction coefficient, which can (possibly) increase with the size of the NANOF; (iv) ferrimagnetism or ferromagnetism, which can also increase the absorption, absorption coefficient, absorption index, or extinction coefficient of the NANOF and / or promote the coupling between the dipole and / or magnetic moment of the NANOF and the dipole and / or magnetic moment of the luminescent substance; or (v) good crystallinity, which can in particular enable the transfer of energy, particles (preferably electrons) between the NANOF and the luminescent substance.

[0140] In one embodiment of the present invention, when the luminescent intensity of the luminescent substance and the molar extinction coefficient of NANOF and the recovery function associated therewith are non-zero, preferably maximized, the luminescent substance associated with NANOF undergoes quenching of luminescent intensity and / or a change in luminescent lifetime compared to the luminescent substance alone.

[0141] In one embodiment of the present invention, the quenching of the luminescence intensity and / or the change in the luminescence lifetime of the luminescent substance associated with NANOF compared to the luminescent substance alone is due to the presence of oxygen atoms or iron atoms in or at the surface of the NANOF.

[0142] In one embodiment of the present invention, when the particles are activated or deactivated, quenching of the luminescence intensity of the luminescent material associated with the NANOF and / or changes in the luminescence lifetime are observed compared to the luminescent material alone. On the other hand, this may be more pronounced when the particles are deactivated than when the particles are activated.

[0143] In one embodiment of the invention, the dissociation of the luminescent substance from the iron oxide nanoparticles and / or the chemical modification of this substance is associated with a change in the luminescence of the luminescent substance. This occurs preferentially when the particles are transformed from an inactivated state to an activated state.

[0144] In one embodiment of the present invention, the dissociation of the luminescent substance from the NANOF suppresses or reduces the quenching of the luminescence intensity, thereby causing an increase in the luminescence intensity.

[0145] In one embodiment of the present invention, the dissociation of the luminescent material from the iron oxide nanoparticles and / or the chemical modification of the luminescent material causes a change in the luminescence of the luminescent material due to: (i) a change (start, increase, stop, or decrease) in the transfer of energy or particles between the luminescent material and the NANOF; (ii) a change in the distance separating the luminescent material from the NANOF; or (iii) a change in the chemical composition or structure of the luminescent material.

[0146] In one embodiment of the present invention, the dissociation of the luminescent material from the NANOF and / or the chemical modification of the luminescent material that causes the luminescence change is: (i) reversible, involving the dissociation of the luminescent material from the NANOF and / or its chemical modification, followed by a return to the initial state of the particle obtained when the particle was deactivated; (ii) irreversible, involving the dissociation of the luminescent material from the NANOF and / or its chemical modification without the luminescent material returning to its initial state; (iii) holistic, involving the entire luminescent material; (iv) partial, involving a part, chemical group, or atom of the luminescent material; (v) related to the movement, translation, oscillation, or vibration of the luminescent material or NANOF; (vi) related to the coupling between the electromagnetic field, the luminescent material and the NANOF; (vii) due to changes in the environment of the particle; or (viii) followed by the diffusion of the luminescent material or a part of it into the particle environment.

[0147] In one embodiment of the invention, the dissociation of the luminescent material from the NANOF and / or the chemical modification of the material is associated with a local luminescence change that occurs when the luminescent material has diffused at a distance preferably less than 100 times, 10 times, or 1 times the size of the NANOF measured from the center of the NANOF. Such local changes in luminescence are typically measured within a measurement time shorter than 100 minutes, 10 minutes, or 1 minute.

[0148] In one embodiment of the invention, at least one parameter related to the dissociation of the luminescent substance and / or the chemical modification of such substance can be measured, such as the amount of luminescent substance that dissociates and / or diffuses, their speed, or the time of diffusion and / or dissociation. It is then possible to establish a relationship between ΔL, ΔL / δt and at least one of these parameters.

[0149] In one embodiment of the invention, a change in the environment of the particle is associated with a change in the luminescence of the luminescent substance. This occurs preferentially when the particle changes from an inactivated state to an activated state.

[0150] In one embodiment of the invention, the change in the environment of the particle and / or the slope at the start of the change can be measured before, after, or during use of the particle, preferably during use of the particle. It is then possible to establish a relationship between ΔL, ΔL / δt, and the change in the environment, such as the pH value, temperature, redox potential, or chemical composition of the environment.

[0151] In one embodiment of the invention, according to the invention, a change in the radiation, such as a change in intensity, energy, frequency, electromagnetic radiation, such as a magnetic field (alternating or non-alternating), or ionizing radiation, to which the particles are exposed is associated with a change in the luminescence of the luminescent substance. This occurs preferentially when the particles are transformed from an inactivated state to an activated state.

[0152] In one embodiment of the invention, the variation of the radiation to which the particles are exposed and / or the slope at the start of this variation can be measured before, after or during, preferably during, the use of the particles according to the invention. It is then possible to establish a relationship between ΔL, ΔL / δt, and the variation of the radiation.

[0153] In one embodiment of the invention, the luminescence change of the probe according to the present invention occurs when the probe interacts with a synthetic or non-synthetic substance, for example, contacts or collides. This can particularly occur during the internalization of the probe into a cell, particularly by applying radiation, preferably ionizing radiation or a magnetic field.

[0154] In one embodiment of the present invention, the change in luminescence of the luminescent substance associated with the iron oxide nanoparticles is different from the change in luminescence of the luminescent substance alone.

[0155] In one embodiment of the present invention, the luminescent substance has at least one luminescent property or undergoes a change in at least one of its properties which is different from the luminescent substance alone, ie the luminescent substance not bound or associated with the particles according to the present invention.

[0156] In one embodiment of the invention, the physicochemical disturbance applied to the particles according to the invention induces a change in at least one luminescence property of the luminescent material, which is different from the change induced by the same disturbance to the luminescent material alone. For example, an increase in the temperature of the particle environment or the application of an alternating magnetic field to the particles can induce the dissociation of the luminescent material from the NANOF and / or the chemical modification of the luminescent material, thereby inducing a change (increase or decrease) in the luminescence of the luminescent material associated with the NANOF, which is different from the luminescence change of the luminescent material alone not bound to the NANOF exposed to the same temperature increase or the same alternating magnetic field.

[0157] In one embodiment of the invention, the luminescent substance associated with the particle is a luminophore, a chromophore or a fluorophore.

[0158] In one embodiment of the present invention, the physicochemical perturbation applied to the particles induces an increase or decrease in luminescence that is 10 times, 10 times, or 10 times greater when the luminescent substance is associated with NANOF than when the luminescent substance is alone and not associated with NANOF. 2 times, 103 times, 10 6 times, 10 10 times, or 10 20 times or 1 / 10, 1 / 10 2 , 1 / 10 3 , 1 / 10 6 , 1 / 10 10 , or 1 / 10 20 .

[0159] In one embodiment of the invention, a physicochemical perturbation applied to a luminescent species associated with a NANOF induces an increase in luminescence, whereas the same perturbation, when applied to a luminescent species alone, not associated with a NANOF, induces a decrease in luminescence.

[0160] In one embodiment of the invention, a physicochemical perturbation applied to a luminescent species associated with a NANOF induces a decrease in luminescence, whereas the same perturbation, when applied to a luminescent species alone, not associated with a NANOF, induces an increase in luminescence.

[0161] In one embodiment of the present invention, a physicochemical perturbation applied to a luminescent substance associated with a NANOF induces a change in luminescence, preferably of at least 10 -10 %, 10 -5 %, 10 -1 %, 1%, 10%, 25%, 50%, 70%, or 90% change, while when the same interference is applied to a single luminescent substance not associated with NANOF, the same interference does not induce a luminescence change, preferably less than 10 -10 %, 10 -5 %, 10 -1 In this case, the percentage of luminescence change can be defined as the ratio between the luminescence measured before and after the perturbation is applied.

[0162] In one embodiment of the present invention, the luminescent substance comprises at least one luminescent chemical atom or group, luminescent molecule, luminescent material, or luminescent monomer or polymer.

[0163] In one embodiment of the present invention, the luminescent substance comprises or consists of at least one substance selected from the group consisting of:

[0164] Luminescent ADN, Luminescent ARN, Alexa fluor, Alexa fluor 350, 405, 430, 488, 500, 514, 532, 546, 548, 555, 568, 594, 610, 633, 647, 660, 669, 680, 700, 750 or 790, ATTO, ATTO 590, 594, 610, 611x, 620, 633, 635, 637, 647, 647N, 655, 655, 665, 680, 700, 725, 740, allophycocyanin (APC), conjugated APC, Cy, APC-Cy7, aminocoumarin, allophycocyanin (APC), APC-Cy7, acridine orange, 7-AAD, azurite, AmCyan1, AcGFP1, Azami-Green, AsRed2, aminocoumarin, allophycocyanin, ethidium bromide, boron-dipyrromethene BODIPY, BODIPY TMR, BODIPY 581 / 591, BODIPY TR, BODIPY-FL, BODIPY 630 / 650 or 650 / 665, ethidium bromide, luminescent cells, calcein, coumarin, anthocyanin Cy, Cy 2, Cy3, Cy3B, Cy 3.5, Cy 5, Cy 5.5 or Cy 7, Cy3.5, organic dyes, cascade blue chromomycin A3, cerulean, CyPet, "DyLightFluor", DyLight 350, 405, 488, 549, 550, 594, 633, 649, 650, 680, 750, 755 or 800, DRAQ5, DAPI, DCFH, DHR, dKeima-Red, Dronpa-Green, DsRed monomer, DsRed2 ("RFP"), Luminase, EBFP, EBFP2, ECFP, Emerald, EYFP, E2-Crimson, Fluorescein, FluoProbe, FluoProbe 594, 647H, 682, 752, 782, FluorX, FAM, Fluorescein FITC, GFPuv, Hydroxycoumarin, Hoechst 33342 or 33258, HcRed1, HyPer, Hydroxycoumarin, Hex, Intercalating luminescent DNA, Propidium iodide, Indo-1Fluo-3, J-Red, Kusabira-Orange, Katushka (TurboFP635), Luciferase, Luminescent lipids, Trivalent lanthanide Ln 3+、YellowLucifer、Lissamine Rhodamine B LDS 751、Lanthanide elements (lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium), megastokes、methoxycoumarin、mithramycin、cell function markers、mCFP、mKeima-Red、mTFP1(cyan)、Midoriishi-Cyan、mCitrine、mBanana、mOrange、mOrange2、mKO、mStrawberry、mRFP1、mCherry、mKate2、mKate(TagFP635)、mPlum、mRaspberry、mNeptune、monochlorodiamine、methoxycoumarin、semiconductor nanocrystals、NBD R-phycoerythrin (PE), aphrodisiac, green fluorescent protein, red fluorescent protein, yellow fluorescent protein, P3, Pacific Blue, PacificOrange, PE-Cy5, PE-Cy7, PerCP, PhiYFP, PhiYFP-m, Red 613, r-phycoerythrin (PE), Rhodamine Red-x, rox, Red 613, Rhodamine, Rhodamine B, SNARF, SYTOX, SYTOX Blue, SYTOX Green, SYTOX Orange, S65A, S65C, S65L, S65T, luminescent substances contained in antibody labeling kits, such as "biotin tag", "fluorescent tag", "immunoprobe" or "Mix-n-Stain", luminescent tissue, Texas Red, TOTO-3, TO-PRO-3, TruRed, TRITC X-Rhodamine, TOTO-1, TO-PRO-1, TO-PRO: cyanine: monomer, thiazole orange, TOTO-3, TO-PRO-3, T-Sapphire, TagBFP, TagCFP, TurboGFP, TagGFP, TagGFP2, Topaz, TurboYFP, TurboRFP, tdTomato DsRed-Express2, TagRFP, TurboFP602, TurboFP635, TRITC, Tamara, Texas Red, TruRed, Venus, YOYO-1Y66H, Y66F, Y66W, YPet, ZsGreen1, ZsYellow1, and substances derived from the above-mentioned luminescent substances or having at least one atom, molecule, or luminescent chemical group identical to the above-mentioned luminescent substances.

[0165] In one embodiment of the invention, the particles according to the invention are associated with a luminescent substance having at least one chemical function selected from the group consisting of carboxylic acids, phosphoric acids, sulfonic acids, esters, amides, ketones, alcohols, phenols, thiol functional groups, amines, ethers, sulfides, anhydrides, acyl halides, amidines, nitriles, hydroperoxides, imines, aldehydes, peroxides, and acidic, basic, oxidized, reduced, neutral, or positively or negatively charged derivatives of these functions.

[0166] In one embodiment of the invention, the luminescent substance has at least one chemical function that allows weak or strong binding to the iron oxide nanoparticles, such as a carboxylic acid function.

[0167] In one embodiment of the invention, the particles according to the invention are associated with a pharmaceutical compound.

[0168] In one embodiment of the invention, the particles according to the invention are associated with a pharmaceutical compound having at least one chemical function selected from the group consisting of carboxylic acids, phosphates, sulfonates, amides, ketones, alcohols, phenols, thiols, amines, ethers, sulfides, anhydrides, acyl halides, amidines, nitriles, hydroperoxides, imines, aldehydes, peroxides, and acidic, basic, oxidized, reduced, neutral, or positively or negatively charged derivatives of these functions.

[0169] In one embodiment of the invention, the drug compound has a chemical functionality that allows for weak or strong binding to the iron oxide nanoparticles, such as a carboxylic acid functionality.

[0170] In one embodiment of the invention, the drug compound is a therapeutic substance, a diagnostic agent, a contrast agent, an anticancer agent, an antibacterial agent, an anti-inflammatory agent, a vaccine, an anesthetic, an analgesic, an antibiotic, an antidepressant, an antidiuretic, an antihistamine, an antihypertensive, an antipyretic, an antiviral, an antitussive, an anxiolytic, a bronchodilator, a diuretic, a laxative, a psychotropic, a sedative, or a vasopressor.

[0171] In one embodiment of the present invention, the pharmaceutical composition comprises an active ingredient and optionally an excipient.

[0172] In one embodiment of the invention, the drug compound has greater efficacy and / or lower toxicity when associated with the particles than when it is alone. This may be due to the fact that it can itself be dissociated from the particles in a controlled manner and activated, for example when it has reached the organism or a desired site of the organism, such as a tumor or tumor cells.

[0173] In one embodiment of the invention, the physicochemical perturbation applied to the particles causes the drug compound to dissociate from the iron oxide nanoparticles.

[0174] In one embodiment, the dissociation of the drug compound from the nanoparticles is controlled by applying a physicochemical perturbation. This may make it possible, in particular, to increase the efficacy of the therapeutic treatment or diagnostic and / or reduce its toxicity.

[0175] In one embodiment of the invention, the physicochemical disturbance is applied to the particles by radiation, preferably ionizing radiation or a magnetic field, more preferably an alternating magnetic field.

[0176] In one embodiment of the invention, said radiation is such that it significantly modifies the NANOF, as defined in the present invention.

[0177] In another embodiment of the present invention, the radiation is a continuous magnetic field of a strength preferably from 0.01 mT to 100 T, from 0.1 mT to 10 T, or from 1 mT to 1 T.

[0178] In one embodiment of the present invention, the radiation is an alternating magnetic field, the intensity of which is preferably 0.01mT to 100T, 0.1mT to 10T, or 1mT to 1T, and preferably includes a frequency of 0.001Hz to 1000MHz, 0.01Hz to 100MHz, 0.1Hz to 10MHz, 1Hz to 1MHz, 10Hz to 500kHz, 100Hz to 500kHz, 1kHz to 250kHz, or 10kHz to 200kHz.

[0179] In one embodiment of the invention, the radiation is of energy greater than 10 -15 Gray, 10 -14 Gray, 10 -13 Gray, 10 -12 Gray, 10 -11 Gray, 10 -10 Gray, 10 -9 Gray, 10 -8 Gray, 10 -7 Gray, 10 -6 Gray, 10 -5 Gray, 10 -4 Gray, 10 -3 Gray, 10 -2 Gray, 10 -1 Gray, 1 Gray, 10 Gray, 10 2 Gray, 10 3 Gray, 10 4 Gray, 10 5 Gray, 10 6 Gray, 10 7 Gray, 10 8 Gray, 10 9 Gray, 10 10Gray, 10 11 Gray, 10 12 Gray, 10 13 Gray, 10 14 Gray, or 10 15 Gray (Gy) of ionizing radiation.

[0180] In one embodiment of the invention, the radiation is of energy less than 10 -15 Gray, 10 -14 Gray, 10 -13 Gray, 10 -12 Gray, 10 -11 Gray, 10 -10 Gray, 10 -9 Gray, 10 -8 Gray, 10 -7 Gray, 10 -6 Gray, 10 -5 Gray, 10 -4 Gray, 10 -3 Gray, 10 -2 Gray, 10 -1 Gray, 1 Gray, 10 Gray, 10 2 Gray, 10 3 Gray, 10 4 Gray, 10 5 Gray, 10 6 Gray, 10 7 Gray, 10 8 Gray, 10 9 Gray, 10 10 Gray, 10 11 Gray, 10 12 Gray, 10 13 Gray, 10 14 Gray, or 10 15 Gray (Gy) of ionizing radiation.

[0181] In one embodiment of the invention, the physicochemical disturbance imposed on the particles according to the invention is due to or is associated with a change in the environment of the particles.

[0182] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.2, 0.1, 0.01, 0.005, 0.001, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, -4 10 -5 10 -6 10-7 10 -8 10 -9 10 -10 10 -11 10 -12 , or 10 -13 A change in pH of 1 pH units.

[0183] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is greater than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.2, 0.1, 0.01, 0.005, 0.001, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, -4 10 -5 10 -6 10 -7 10 -8 10 -9 10 -10 10 -11 10 -12 , or 10 -13 A change in pH of 1 pH units.

[0184] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 10% of the change in the environment or at least one substance contained in the environment. 10 ℃, 10 9 ℃, 10 8 ℃, 10 7 ℃, 10 6 ℃, 10 5 ℃, 10 4 ℃, 10 3 ℃、500℃、250℃、150℃、100℃、50℃、40℃、20℃、30℃、10℃、5℃、1℃、0.1℃、0.01℃、0.001℃、10 -4 ℃, 10 -5 ℃, 10 -6 ℃, 10 -7 ℃, 10 -8 ℃, 10 -9 ℃, 10 -10 ℃, 10 -11 ℃, 10 -12 ℃, or 10 -13 ℃ temperature change.

[0185] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is greater than 10% of the change in the environment or at least one substance contained in the environment. 10 ℃, 10 9 ℃, 10 8 ℃, 10 7 ℃, 10 6 ℃, 10 5 ℃, 10 4 ℃, 10 3 ℃、500℃、250℃、150℃、100℃、50℃、40℃、20℃、30℃、10℃、5℃、1℃、0.1℃、0.01℃、0.001℃、10 -4 ℃, 10 -5 ℃, 10 -6 ℃, 10 -7 ℃, 10 -8 ℃, 10 -9 ℃, 10 -10 ℃, 10 -11 ℃, 10 -12 ℃, or 10 -13 ℃ temperature change.

[0186] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 10% of the change in the environment or at least one substance contained in the environment. 3 Volts, 500 Volts, 100 Volts, 10 Volts, 5 Volts, 4 Volts, 3 Volts, 2 Volts, 1 Volt, 0.5 Volts, 0.4 Volts, 0.3 Volts, 0.2 Volts, 0.1 Volts, 0.01 Volts, 0.001 Volts, 10 -4 Volt, 10 -5 Volt, 10 -6 Volt, 10 -7 Volt, 10 -8 Volt, 10 -9 Volt, 10 -10 Volt, 10 -11 Volt, 10 -12 Volts, or 10 -13 The standard change in potential of volts.

[0187] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 10% of the change in the environment or at least one substance contained in the environment. 3 Volts, 500 Volts, 100 Volts, 10 Volts, 5 Volts, 4 Volts, 3 Volts, 2 Volts, 1 Volt, 0.5 Volts, 0.4 Volts, 0.3 Volts, 0.2 Volts, 0.1 Volts, 0.01 Volts, 0.001 Volts, 10-4 Volt, 10 -5 Volt, 10 -6 Volt, 10 -7 Volt, 10 -8 Volt, 10 -9 Volt, 10 -10 Volt, 10 -11 Volt, 10 -12 Volts, or 10 -13 The standard change in potential of volts.

[0188] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is greater than 10 10 , 10 9 , 10 8 , 10 7 , 10 6 , 10 5 , 10 4 , 10 3 , 500, 250, 150, 100, 50, 40, 20, 30, 10, 5, 1, 0.1, 0.01, 0.001, 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , or 10 -13 A change in the concentration of at least one substance of the environment in mol / L, micromol / L, nanomol / L, mole / ml, micromol / ml, nanomol / ml, mole / m3, mole / dm3, mole / cm3, or mole / mm3.

[0189] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 10 10 , 10 9 , 10 8 , 10 7 , 10 6 , 10 5 , 10 4 , 10 3 , 500, 250, 150, 100, 50, 40, 20, 30, 10, 5, 1, 0.1, 0.01, 0.001, 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8, 10 -9 , 10 -10 , 10 -11 , 10 -12 , or 10 -13 A change in the chemical composition of at least one substance of the environment on a mol / L, micromol / L, nanomol / L, mole / ml, micromol / ml, nanomol / ml, mole / m3, mole / dm3, mole / cm3, or mole / mm3 basis.

[0190] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is at least 1, 2, 3, 4, 5, 10, 25, 50, 100, 500, 10 3 10 4 10 5 10 6 10 7 10 8 10 9 Species, or 10 10 A change in a substance, wherein the change can be a chemical or structural change and / or the appearance or disappearance of one or more substances from the environment.

[0191] In one embodiment of the invention, the physicochemical disturbance imposed on the particle or the change in the environment of the particle according to the invention is less than 2, 3, 4, 5, 10, 25, 50, 100, 500, 10 3 10 4 10 5 10 6 10 7 10 8 10 9 Species, or 10 10 A change in the chemical composition of a substance, wherein the change can be a chemical or structural change and / or the appearance or disappearance of one or more substances in the environment.

[0192] In one embodiment of the invention, the physicochemical disturbance applied to the particle induces a change in the state of the particle, which change can be defined as: (i) the pH value of the particle increases or decreases by more than or less than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.2, 0.1, 0.01, 0.005, 0.001, 10 -4 10 -5 10 -6 10 -7 10-8 10 -9 10 -10 10 -11 10 -12 , or 10 -13 pH units; (ii) the temperature of the particles increases or decreases by more than or less than 10 10 ℃, 10 9 ℃, 10 8 ℃, 10 7 ℃, 10 6 ℃, 10 5 ℃, 10 4 ℃, 10 3 ℃、500℃、250℃、150℃、100℃、50℃、40℃、20℃、30℃、10℃、5℃、1℃、0.1℃、0.01℃、0.001℃、10 -4 ℃, 10 -5 ℃, 10 -6 ℃, 10 -7 ℃, 10 -8 ℃, 10 -9 ℃, 10 -10 ℃, 10 -11 ℃, 10 -12 ℃, or 10 -13 ℃; (iii) the charge of the particles increases or decreases by more than or less than 10 3 Volts, 500 Volts, 100 Volts, 10 Volts, 5 Volts, 4 Volts, 3 Volts, 2 Volts, 1 Volt, 0.5 Volts, 0.4 Volts, 0.3 Volts, 0.2 Volts, 0.1 Volts, 0.01 Volts, 0.001 Volts, 10 -4 Volt, 10 -5 Volt, 10 -6 Volt, 10 -7 Volt, 10 -8 Volt, 10 -9 Volt, 10 -10 Volt, 10 -11 Volt, 10 -12 Volts, or 10 -13 volts; or (iv) the number of atoms contained in the particles increases or decreases by more than or less than 10 100 10 50 10 20 10 10 10 3 , 10, 5, or 1 atom.

[0193] The present invention also relates to an in vitro method for detecting changes in luminescence, said method comprising the step of detecting changes in luminescence of the particles according to the present invention.

[0194] In one embodiment of the in vitro method for detecting a change in luminescence according to the invention, said change in luminescence is measured during a time interval shorter than 1 hour, 1 minute, or 1 second.

[0195] In another embodiment of the in vitro method for detecting a change in luminescence according to the present invention, the particle is surrounded by a luminescence detector, preferably within a radius of less than 1 cm measured from the center of the particle. 3 , 1mm 3 , or 0.1mm 3 The luminescence change is measured locally at a distance of .

[0196] In another embodiment of the in vitro method for detecting a change in luminescence according to the present invention, the change in luminescence is a continuous increase in the luminescence of the luminescent substance, preferably lasting at least 1 second, 1 minute, or 1 hour.

[0197] In another embodiment of the in vitro method for detecting a change in luminescence according to the present invention, dissociation of the luminescent substance from NANOF is detected.

[0198] In another embodiment of the present invention, the method for detecting luminescence changes according to the present invention is applied to detect at least one physiological parameter, preferably temperature, pH value, environment of the iron oxide nanoparticles.

[0199] In another embodiment, the method for detecting changes in luminescence according to the invention is applied to detect radiation as defined above.

[0200] In another embodiment of the in vitro method for detecting changes in luminescence according to the invention, the luminescence intensity of the particles according to the invention is increased by applying radiation as defined above.

[0201] In another embodiment of the in vitro method for detecting changes in luminescence according to the present invention, the luminescence intensity of the luminescent substance is increased by changing the environment of the particles.

[0202] The present invention also relates to a device, comprising:

[0203] - a particle according to the invention, wherein the compound is a luminescent substance, and

[0204] -System for detecting changes in luminescence.

[0205] Preferably, the device according to the invention comprises a luminescent substance associated with the particles, the luminescence of which is spontaneous, and a detection system making it possible to measure changes in the luminescence of the luminescent substance associated with the particles.

[0206] It is also preferred that the device according to the invention comprises a luminescent substance associated with the particles, the luminescence of which is induced by a first excitation source and detected by a detection system which makes it possible to measure changes in the luminescence of the luminescent substance associated with the particles.

[0207] The first excitation source induces the luminescence of the luminescent material associated with the particle. It can be a monochromatic or polychromatic light source operated continuously or by pulse operation. It preferably produces single-photon excitation or multi-photon excitation and emits at a wavelength preferably of 300nm to 1500nm. It can be a laser, such as a crystal laser, a Yag laser emitting at 1064nm, a dye laser, a gas laser, a laser diode, a free electron laser or a fiber laser. It can also be a lamp, such as a halogen lamp. The detection system is a hardware or hardware group for detecting the luminescence change of the luminescent material associated with the particle. It can include equipment for selecting the wavelength of the radiation emitted by the probe, such as a wavelength spectrum analyzer, a spectrometer, a photosensitive sensor (its resistance changes with the change of the intensity of the captured light radiation) (such as a photoelectric unit, a photovoltaic unit or a photoresist unit), a scintillator, a photomultiplier tube, a channel multiplier, a photographic sensor, a detector for dispersive energy analysis, an infrared radiation detector, and a photodetector.

[0208] In one embodiment of the invention, the device according to the invention further comprises a second excitation source, such as a radiation source, preferably an ionizing radiation source, or a magnetic field source, preferably an alternating magnetic field. This second excitation source may make it possible to induce a change in the luminescence of the luminescent substance associated with the particle.

[0209] In another embodiment of the present invention, the device according to the present invention includes an optical component, which contains at least one material for transmitting an optical signal, such as an optical fiber, a microscope, a lens, which can be used to transmit light from an excitation source to a luminescent substance associated with the particle, to excite the luminescent substance, to collect the luminescence of the luminescent substance and / or to transmit the luminescence of the luminescent substance from the luminescent material to a detection system.

[0210] In another embodiment of the present invention, the device according to the present invention comprises one or more detection systems for detecting at least one parameter of the particles according to the present invention, the presence of low frequency / natural abundance substances / isotopes, characteristics of radiation (e.g. frequency, intensity, energy of electromagnetic radiation or ionizing radiation), transition metals, or substances of medical interest. The detection system according to the present invention may contain TIMS (solid source thermal ionization mass spectrometer) or ICP-MS (inductively coupled plasma mass spectrometer), radioisotope detectors (also known as radioisotopes) (e.g. nuclear radiation detection instruments), gas, semiconductor, inorganic scintillator, organic liquid scintillator or solid organic scintillation detector, photomultiplier tube, Geiger counter, physiological parameter probe (e.g. temperature or pH probe), or magnetic field detection probe.

[0211] In one embodiment of the invention, the device according to the invention allows the particles according to the invention to be converted from an inactivated state into an activated state, wherein the activation can in particular lead to a change in at least one luminescence property of the luminescent substance.

[0212] In one embodiment of the invention, activation of the device is performed when the particles according to the invention are activated.

[0213] In one embodiment of the invention, the activation of the device according to the invention and / or of the particles according to the invention causes a change in at least one luminescence property of the particles, which change is measurable in the absence of a second excitation source and the device need not include a second excitation source. This may occur, for example, when a change in the environment of the particles induces a change in the luminescence.

[0214] In one embodiment of the invention, activation of the device according to the invention and / or of the particles according to the invention causes a change in at least one luminescence property of the particles which cannot be measured in the presence of a second excitation source and which is then comprised in the device.

[0215] In one embodiment of the invention, the device according to the invention functions according to at least one of the following sequences: luminescence of a luminescent substance associated with a particle (sequence 1), a change in luminescence of a luminescent substance associated with a particle (sequence 2), and detection of the change in luminescence (sequence 3). Typically, these sequences are consecutive and sequence 3 is performed almost simultaneously with sequence 1 and / or sequence 2, because it is desirable to measure the change in luminescence immediately after exciting the particle to avoid luminescence losses. The time separating sequence 3 from sequence 1 and / or sequence 2 may be shorter than 1 minute, preferably shorter than 1 second, and most preferably shorter than one hundredth of a second. Furthermore, it may occur that sequence 1 and sequence 2 are performed simultaneously, for example when radiation induces both luminescence and a change in luminescence of the particle, or that sequence 1 is performed after sequence 2.

[0216] In one embodiment of the invention, particles according to the invention and / or devices according to the invention can be used to measure the presence of the particles and their spatial distribution. In this case, radiation as defined above can be used to increase the luminescence of the particles and make it easier to detect by the device according to the invention. In addition, an optical system can be used to measure the spatial distribution of particles, the optical system comprising an optical fiber that moves horizontally on a sample containing particles according to the invention. It is then possible to establish a relationship between the spatial luminescence variation of particles and the particle distribution in a given medium.

[0217] The present invention also relates to a pharmaceutical composition comprising at least the particles according to the present invention, optionally in combination with a pharmaceutically acceptable vehicle.

[0218] In a preferred embodiment of the pharmaceutical composition according to the present invention, the compound is a pharmaceutical compound.

[0219] According to another aspect of the invention, the particles according to the invention are used as a medicament, in particular for medical, veterinary or phytochemical applications.

[0220] The present invention also relates to a method for treating a disease including cancer in an individual, wherein a therapeutically active amount of particles according to the present invention is administered to the individual.

[0221] The invention furthermore relates to a diagnostic composition comprising at least the particles according to the invention.

[0222] In a preferred embodiment of the diagnostic composition according to the invention, said compound is a luminescent substance as defined above.

[0223] In one embodiment of the invention, the particles according to the invention are used as diagnostic agents or in vivo probes.

[0224] The invention also relates to a diagnostic method, wherein the particles according to the invention are administered to an individual.

[0225] The invention also relates to a medical device comprising at least the particles according to the invention, wherein the compound is preferably a pharmaceutical compound or a luminescent substance.

[0226] In one embodiment of the present invention, the medical device according to the present invention is used in medical, veterinary or phytotherapy applications, in particular for the treatment of diseases, such as cancer.

[0227] The invention also relates to a cosmetic composition comprising at least the particles according to the invention, wherein the compound is a cosmetically active ingredient, optionally in combination with a cosmetically acceptable vehicle.

[0228] According to another aspect of the invention, the particles according to the invention are used as cosmetic agents.

[0229] According to the invention, the particle may be a medical device, a medicament, a cosmetic composition, a therapeutic substance, a contrast agent or a diagnostic agent.

[0230] According to another aspect of the invention, the particles according to the invention comprising a luminescent compound are used as probes to detect changes in the luminescence or in the properties of the NANOF, the luminescent substance and / or the particle environment.

[0231] In some embodiments, the luminescent compound is a luminescent substance.

[0232] In one embodiment of the present invention, the particles according to the present invention comprising a luminescent compound are used as probes to detect the dissociation of the luminescent substance from the NANOF and / or the chemical modification of the NANOF.

[0233] In one embodiment of the present invention, the particles according to the present invention comprising a luminescent compound are used as probes for detecting radiation as defined according to the present invention.

[0234] In one embodiment of the present invention, the probe according to the present invention comprising particles associated with a luminescent compound can be used in vivo, ex vivo or in vitro, in the context of medical research or non-medical research (in particular treatment, diagnosis), and / or inside, on or outside a substance (preferably a biological substance).

[0235] In one embodiment of the invention, the particles according to the invention comprising a luminescent compound are used as probes to detect a specific physiological state different from normal, preferably caused by a disease or a medical procedure.

[0236] According to one embodiment of the present invention, the particle according to the present invention is used as a probe to detect a specific physiological state different from the physiological state of the subject in its normal state. This specific physiological state can be caused by disease, medical operation, exposure of the organism to material or radiation, fever, cancer, immune system reaction. It can also be any state reached by the subject during operation or medical intervention, treatment, diagnosis, surgery, biopsy process, cytological puncture, endoscopy, MRI, scanner, colonoscopy, scintigraphy, PET-SCAN, X-ray, chemotherapy, hyperthermia (such as magnetic hyperthermia). For this reason, the particle can be made to interact, contact, associate or be applied to the organism in a specific physiological state with the organism. This can induce changes in the luminescence of the particle, which are different from the changes observed when the body is in its normal state and therefore allow the detection of this specific physiological state. For example, the particle can be introduced into the tumor of a patient undergoing hyperthermia treatment and the measured luminescence changes may be different from the changes observed in the absence of treatment, thereby indicating the activation of the treatment.

[0237] In one embodiment of the invention, the particles according to the invention comprising a luminescent compound are used as probes to detect the association, labelling, targeting, delivery of particles, NANOFs or luminescent substances.

[0238] According to one embodiment of the present invention, the particles according to the present invention are used for the labeling, targeting, and delivery of detection substances. In order to label, the particles can be associated with the substance. In order to target a substance, the particles can be transported or guided to the substance, for example, by connecting the particles to antibodies that specifically recognize the substance or applying a magnetic field oriented toward the substance. In order to deliver a substance, the particles can be associated with the substance and delivered to a given location. The labeling, targeting, and delivery of the substance can then be observed from the characteristic luminescence changes that occur during the labeling, targeting, and delivery.

[0239] In one embodiment of the invention, the particles according to the invention comprising a luminescent compound are used as probes to detect the removal of a mixture, particles from a medium.

[0240] The invention further relates to the purposes of the particle according to the present invention as probe for detecting the removal of some substances from material, organism, medium, particularly under the background of detoxification or for separating and purifying these substances. For this reason, the particle can be connected with these substances and can use for example magnetic field that the complex thus formed is removed from material, organism, medium. This removal can be observed from the specific luminescent change that occurs during the removal.

[0241] The invention still further relates to the purposes of the mixing of particle according to the present invention as probe for monitoring some substance and material, organism or medium.For this reason, the probe can be connected with these substances and magnetic field can be used to mix the thus formed complex with material, organism or medium.This mixture can be observed from the specific luminescent change that occurs during mixing.

[0242] In one embodiment of the invention, the particles according to the invention comprising a luminescent compound are used as probes to detect specific conditions different from normal conditions, such as high levels of pollution, radioactivity, toxicity.

[0243] According to one embodiment of the invention, the particles according to the invention are used as probes to detect specific conditions that differ from normal conditions, for example conditions caused by levels of certain parameters, such as radioactivity, contamination, radiation, toxicity, that differ from, preferably are higher than normal. To this end, the particles can be placed in locations with different levels of these parameters, preferably higher than normal. This can induce changes in luminescence that differ from the changes observed in locations with normal levels of these parameters and thus allow the detection of different levels of these parameters, preferably higher than normal levels.

[0244] The invention also relates to the use of the particles according to the invention as probes for detecting the efficacy of transfection or magnetofection. In this case, DNA or RNA can be associated with the particles and transfected into cells in the presence or absence of a magnetic field. The efficacy of these mechanisms can be confirmed by specific luminescence changes that occur during transfection or magnetofection.

[0245] The present invention further relates to the use of a particle according to the invention as a probe for detecting internalization of said particle, for example in a cell or an organelle, wherein the internalization can be observed from specific luminescence changes occurring during the internalization.

[0246] The invention also relates to the use of the particles according to the invention as probes and / or for detecting the dissociation of a substance from said particles and / or its diffusion, in particular after application of radiation or a change in the environment of said particles. The dissociation of the substance and / or its diffusion can be highlighted from the specific luminescence changes it (they) induces.

[0247] In one embodiment of the invention, the particles or luminescent substances according to the invention undergo a luminescence variation which can be used to detect at least one of the following parameters: (i) a change in the environment of the particles, NANOF or luminescent substance as defined in the invention; (ii) radiation as defined in the invention; (iii) dissociation of a compound from a NANOF and / or diffusion of a compound; or (iv) distribution of the particles. In order to enable this detection, a relationship can be established between the luminescence variation and at least one of these parameters, which is achieved by using a known or determined relationship or by using at least one detection system making it possible to measure at least one of these parameters.

[0248] The present invention also relates to particles comprising at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one compound, wherein after a physicochemical disturbance is applied to the particles, the compound dissociates from the iron oxide nanoparticle and / or is itself chemically modified, and the iron oxide nanoparticles are synthesized by bacteria, preferably magnetotactic bacteria, and do not contain organic or carbonaceous materials derived from the bacteria. In this case, the particles may contain mineral materials derived from the bacteria.

[0249] In one embodiment of the invention, when the particles contain less than 90%, 75%, 50%, 25%, 15%, 10%, 5%, 3%, 1%, 10%, -1 %, 10 -2 %, 10 -3 %, 10 -4 %, 10 -10 %, or 10 -20 % of carbonaceous material or organic material derived from living organisms or the bacteria, the particles do not contain such carbonaceous material or organic material, wherein the percentage may correspond to the mass of the carbonaceous material or organic material derived from living organisms or the bacteria contained in the particles divided by the total mass of the particles.

[0250] In one embodiment of the invention, when the particles are pyrogen-free or when the particles contain less than 10 3 The particles do not contain carbonaceous material or organic material derived from living organisms or the bacteria when the endotoxin concentration is less than 1, 1000, 500, 100, 50, 10, or 5 endotoxin units.

[0251] In one embodiment of the invention, when the particles are not derived from genetically modified bacteria, the particles do not contain any organic or carbonaceous material derived from the bacteria, wherein the genetic modification is aimed at producing the synthesis of a luminescent substance.

[0252] The present invention also relates to particles comprising at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle associated with at least one luminescent substance, wherein the luminescent substance dissociates from the iron oxide nanoparticle and / or undergoes chemical modification itself after a physicochemical disturbance is applied to the particles, and the luminescent substance has a size that is at most 1 / 100 of the size of the nanoparticle, and the luminescent substance is in direct contact with the nanoparticle, and the particles do not contain any carbonaceous material or organic material derived from a living organism.

[0253] In one embodiment of the present invention, the size of the luminescent substance is at most 1 / 2, 1 / 5, 1 / 10, 1 / 10 of the size of the nanoparticle. 2 , 1 / 10 3 , 1 / 10 4 , 1 / 10 5 , or 1 / 10 10 , which may make it possible to bind or associate a large amount of luminescent substances with the nanoparticles.

[0254] In one embodiment of the present invention, the luminescent substance has a small size, preferably less than 10 nm or 1 nm. or or small molecular weight, preferably less than 10 6 g / mol, 10 5 g / mol, 10 4 g / mol, 10 3 g / mol, 10 2 g / mol, or 10 g / mol, which may make it possible to combine or associate a large amount of luminescent substances with the nanoparticles.

[0255] In one embodiment of the present invention, the luminescent substance is combined or associated with the nanoparticles through at least one substance, the size of which is less than 10nm or 1nm, or Or molecular weight less than 10 6 g / mol, 10 5 g / mol, 10 4 g / mol, 10 3 g / mol, 10 2 g / mol, or 10 g / mol, which may make it possible to combine or associate a large amount of luminescent substances with the nanoparticles.

[0256] In one embodiment of the present invention, it is preferred to avoid using particles larger than 10 μm or 1 μm, 100 nm, 10 nm or 1 nm, or The luminescent substance may be bound or associated with the nanoparticle by binding to or associating the luminescent substance with the nanoparticle by binding to or associating the luminescent substance with the nanoparticle with certain large structures, such as biological substances, proteins, lipids, DNA, RNA, or macromolecules, such as dendrimers. This type of structure, on the one hand, limits the number of luminescent substances that can be bound or associated with the nanoparticle, and on the other hand, may produce a significant distance between the luminescent substance and the nanoparticle, preferentially greater than 10 μm or 1 μm, 100 nm or 10 nm, and thus interfere with the performance of the probe or particle, including quenching of the luminescence of the luminescent substance when the luminescent substance is close to the nanoparticle.

[0257] In one embodiment of the present invention, the surface / volume ratio of the nanoparticles is high enough to allow the compound or luminescent substance to associate, and the surface / volume ratio is preferably greater than 10. -10 nm -1 , 10 -5 nm -1 , 10 -3 nm -1 , 10 -1 nm -1 , 0.5nm -1 , or 0.9nm -1 .

[0258] In another embodiment of the present invention, the surface / volume ratio of the nanoparticles is not too high thereby preventing the nanoparticles from losing their ferrimagnetic or ferromagnetic properties, preferably less than 10 3 nm -1 , 10 2 nm -1 , 10nm -1 , 1nm -1 , 0.1nm -1 , or 0.01nm -1 .

[0259] In one embodiment of the invention, at least 2, 5, 10, 10 2 10 3 10 4 10 5 , or 10 10The number of luminescent materials associated or bound to the nanoparticle or each nanoparticle is preferably associated or bound to each nanoparticle on average. In some cases, the number of luminescent materials associated or bound to each nanoparticle on average can be estimated by measuring the number of luminescent materials not associated with the nanoparticles contained in a given volume when the particles are activated or inactivated (preferably when the particles are activated) divided by the number of nanoparticles not associated with the luminescent materials contained in the same volume when the particles are activated or not activated (preferably when the particles are activated).

[0260] In one embodiment of the invention, the particles comprising a luminescent substance associated or bound to the nanoparticles have a SAR, a coercive force, a ratio between the residual magnetization and the saturation magnetization, or a saturation magnetization that is at least equal to or equal to 10 times the SAR, the coercive force, the ratio between the residual magnetization and the saturation magnetization, or the saturation magnetization of the nanoparticles, respectively. -4 times, 10 -3 times, 10 -2 times, 10 -1 times, 1 times, 1.001 times, 1.01 times, 1.2 times, 1.5 times, 2 times, 5 times, 10 times, or 100 times.

[0261] In one embodiment of the invention, at least two of the following environments may be equivalent: chemical environment, particle environment, NANOF environment, compound environment, probe's environment, particle's natural environment, radioactive environment, biological environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0262] Figure 1 : A schematic diagram showing the manufacture and use of the particles. Under the action of ionizing radiation or a change in the environment of the particles, the compounds dissociate from the NANOF and / or are chemically modified.

[0263] Experimental Example:

[0264] Abbreviations meaning:

[0265] In the description of the experimental examples, MC represents a magnetosome chain extracted from non-luminescent magnetotactic bacteria grown in the absence of rhodamine B, MCR400 and MCR20 represent magnetosome chains extracted from magnetotactic bacteria, wherein the bacteria are synthesized in the presence of 400 μM rhodamine B and 20 μM rhodamine B. BNF-DiI and MC-DiI represent ordinary BNF-starch (Micromod 10-00-102) and magnetosomes in chain form prepared in the presence of DiI (Sigma 42364; 1,1'-di-octadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate), respectively. The supernatant of MCR400, BNF-DiI and MC-DiI is the liquid removed after attracting various iron oxide nanoparticles in suspension using a 0.6 T neodymium magnet.

[0266] Example 1: Synthesis and characterization of MC and MCR400

[0267] Suspensions of MC and MCR400 were prepared according to the following experimental scheme. Magnetotactic bacteria Magnetospirillum magneticum AMB-1 was obtained from ATCC (ATCC 700274) and the bacteria were cultured in a slightly modified liquid medium compared to MSGM medium (ATCC medium 1653) at 30°C under microanaerobic conditions. In 1 liter, this medium contained 0.68g of potassium dihydrogen phosphate, 0.85g of sodium succinate, 0.57g of sodium tartrate, 0.083g of sodium acetate, 225μl of resazurin 0.2%, 0.17g of sodium nitrate, 0.04g of L-ascorbic acid, 2ml of 10mM quinic acid iron solution, 10ml of Woolf vitamin solution and 5ml of Woolf mineral solution. In order to prepare MC suspensions, rhodamine solution was not added to the medium. In order to prepare the MCR400 suspension, 100mL of 4mM Rhodamine B solution was introduced into the culture medium of bacteria. In order to prepare the MCR20 suspension, 5mL of 4mM Rhodamine B solution was introduced into the culture medium of bacteria. The pH value of the culture medium was adjusted to 6.85 using 1M sodium hydroxide solution. During the stationary phase, bacteria were collected and tangential flow filtration column (mPES, 500KDa) was used to concentrate the bacteria at a flow rate of 950 ml / min, and then the phosphate buffer solution (pH 7.4) (137mM NaCl, 2.7mM KCl, 10mM Na 2 HPO 4 , 1.76 mM KH 2 PO 4) for 5 times for 30 minutes. The bacteria were collected by centrifugation at 4000 rpm for 1 hour, the supernatant was removed and the bacteria were resuspended in 50 mM Tris-HCl buffer solution (pH 7.4) and diluted to obtain an optical density of 5 at 600 nm. To extract the magnetosome chains, the suspension was sonicated at 30 W (2 s pulse and 1 s pulse interval) at 5°C for 60 minutes to lyse the cell membrane and thus separate the magnetosome chains from the bacteria. After sonication, the magnetosome chains were magnetically separated from the cell debris using a neodymium magnet. The supernatant containing the cell debris was removed and the magnetosome chains were magnetically washed five times with 50 mM Tris-HCl buffer solution (pH 7.4) and washed with Wash them 15 times with water magnetic field. Finally, resuspend them in A suspension of MCR400, MCR20 or MC was obtained in sterile water.

[0268] The absorption spectrum of the MCR400 suspension at an iron concentration of 40 μg / mL reveals the association of Rhodamine B with MCR400. The spectrum shows a shoulder peak at 550 nm, which is not present in the absorption spectrum of MC and corresponds to the maximum absorption wavelength of Rhodamine B. In addition, the supernatant of the suspension of MCR400 has a small amount of absorption at this wavelength. This confirms the fact that the absorption of Rhodamine B in the suspension of MCR400 comes from Rhodamine B associated with the magnetosomes. The association can also be revealed by the fluorescence emission spectrum of MCR400. In fact, the suspension of MCR400 excited at 405 nm has a maximum fluorescence intensity at 569 nm, which is slightly lower than the wavelength corresponding to the maximum fluorescence intensity of Rhodamine B, i.e. 576 nm. This difference can be explained by the association of Rhodamine B with the magnetosome chain, which will slightly change the emission wavelength of MCR400 compared to Rhodamine B. In addition, our measured infrared spectrum of MCR400 reveals the presence of characteristic deformation and vibration bands of MC (1650 cm -1 Amide I at 1530 cm -1 Amide II at 1050 cm -1 and 1250cm -1 Lipopolysaccharide (LPS) or phospholipids at 580 cm -1 magmacite) and the characteristic deformation and vibration bands of rhodamine B (1700 cm -1 Carboxylic acid at 1475 cm -1 and 680cm -1 Olefins at 1250cm -1 and 1175cm -1 Ether, 1125cm -1These results therefore seem to confirm that Rhodamine B is adsorbed at the surface of the magnetosomes or complexed with the magnetosomes, probably because of the carboxylic acid function of Rhodamine B. The experiments also made it possible to determine the number of Rhodamine B molecules present on or in each magnetosome. To this end, MCR400 was placed in the presence of hydrochloric acid (pH = 2) in order to release Rhodamine B from the magnetosomes and convert the iron oxide into iron (III) ions and iron (II) ions. The fluorescence spectrum of MCR400 treated in this way shows that Rhodamine B is fully released from the magnetosomes, since the luminescence peak is at 582 nm (emission wavelength of free Rhodamine B in solution (pH 2)) and not at 568 nm (emission wavelength of Rhodamine B associated with magnetosomes in MCR400). Based on the luminescence intensity of rhodamine B released from the magnetosomes (about 0.56 au), the rhodamine B concentration was estimated to be 750 nmol / L from a calibration curve representing the luminescence variation of rhodamine B alone in a solution (pH = 2) as a function of rhodamine B concentration. 14 RhB molecules. It is known that the concentration of magnetosomes in the suspension is about 166 μg / mL and the mass of magnetosomes is 6.25×10 - 16 g, we deduced that the number of magnetosomes in 1 mL is 26.5×10 10 magnetosomes and the number of Rhodamine B molecules associated with the magnetosomes in MCR400 is estimated to be about 178±4. In other words, 178±4 Rhodamine B molecules are associated with each magnetosome on average.

[0269] We can conclude from this example:

[0270] (i) We have proposed a method for producing MCR400.

[0271] (ii) We have confirmed the association of rhodamine B with magnetosomes in MCR400.

[0272] (iii) MCR400 is characterized by an absorption peak at a wavelength similar to that of Rhodamine B and an emission peak at a different emission wavelength that differs from Rhodamine B by about 7 nm-10 nm.

[0273] Example 2: Synthesis and properties of MC-Dil and BNF-Dil

[0274] The suspensions of MC-DiI and BNF-DiI are prepared starting from suspensions of MC and BNF-starch, respectively, which have been previously concentrated to 25 mg / mL (in terms of iron oxide) by centrifugation at 14,000 rpm for 30 minutes at 5°C. A stock solution of 5 mg / mL of DiI is first prepared in previously distilled anhydrous ethanol. Then 5 mg of the suspension of MC and BNF-starch of iron oxide are introduced into 200 μL of distilled anhydrous ethanol and then mixed with 2.5 mg of DiI from the stock solution. The mixture is sonicated at 30 W for 10 minutes (pulses of 2 seconds) under light protection and at a temperature comprised between 5°C and 10°C. A volume of 1 ml of sterile Water was added to the sonicated suspension, and the sample was then kept under magnetic stirring for 1 hour at room temperature and protected from light. Subsequently, the mixture was stored at 5°C for 24 hours. The mixture was sonicated at 30W for 10 minutes (pulse 2 seconds) at a temperature of 5°C to 10°C in the dark. The suspension of MC-DiI and BNF-DiI was concentrated by centrifugation at 14,000 rpm for 30 minutes at 5°C, then washed 5 times with 0.5% ethanol and washed with sterile Wash them 15 times with water. Finally, resuspend them in in sterile water to obtain a suspension of MC-DiI or BNF-DiI.

[0275] The absorption spectrum of DiI has two maxima at 560nm and 520nm. BNF-DiI shows three absorption peaks at different wavelengths of 595nm, 535nm and 490nm. BNF-DiI supernatant does not absorb at the absorption wavelength of DiI. The emission spectrum of DiI excited at 530nm shows two peaks with maximum intensity at 640nm and 591nm. For the absorption spectrum, the emission spectrum of BNF-DiI excited at 530nm (its peak with maximum intensity is at 670nm and 600nm) and the emission spectrum of DiI are observed to shift the spectrum. In addition, BNF-DiI supernatant does not produce luminous intensity at the emission wavelength of DiI. The absorption and emission spectrum shifts observed between BNF-DiI and DiI show its changes when DiI is associated with BNF-starch. The lack of absorption and emission in BNF-DiI supernatant shows that there is no DiI in the supernatant and the association of DiI with BNF in BNF-DiI.

[0276] MC-DiI showed relatively similar behavior to BNF-DiI with two absorption peaks at 570 nm and 520 nm and two emission peaks at 680 nm and 600 nm. For BNF-DiI, the differences observed between the absorption and emission peaks of MC-DiI and those of DiI indicate that DiI is altered when it associates with MC. The absence of absorption and emission in the MC-DiI supernatant indicates that DiI is associated with MC in MC-DiI.

[0277] We can conclude from this example:

[0278] (i) We present a method for producing BNF-DiI and MC-DiI.

[0279] (ii) We highlighted the association of DiI with BNF and MC.

[0280] (iii) BNF-DiI and MC-DiI are characterized by absorption and emission peaks at wavelengths different from those of DiI.

[0281] Example 3: Using MCR400 as a temperature probe.

[0282] The solution of the Rhodamine B of concentration 5 μM and pH 7 contained in the volume of 800 μL is excited at 405nm and heated at different temperatures for 30 minutes using a dry hot bath. When the temperature of the solution is increased to 80 ℃ from 20 ℃, the wavelength corresponding to the maximum luminescence of Rhodamine B remains relatively unchanged, and the maximum luminous intensity of Rhodamine B is strongly reduced by 73%, which is consistent with previously disclosed results (Journal of Luminescence, Vol. 27, p. 455 (1982)). The MCR400 suspension of 250 μg / mL iron oxide concentration and pH 7 contained in the volume of 800 μl is excited at 405nm and heated at different temperatures for 30 minutes using a dry hot water heating bath. The MCR400 excited at 405nm produces maximum luminous intensity at 569nm. The behavior of MCR400 is different from that of Rhodamine B. In fact, at 20°C to 80°C, the maximum of the luminescence peak was observed at 569 nm and the luminescence intensity of MCR400 increased dramatically by 66%. The luminescence intensity of Rhodamine B will be quenched initially when Rhodamine B associates with magnetosomes and will increase as Rhodamine B dissociates from magnetosomes under the effect of increasing temperature.

[0283] The dissociation of Rhodamine B from MCR400 by heating was confirmed by absorption measurement. A MCR400 suspension of 40 μg / mL iron oxide concentration was heated at pH 7 for a period of 0 to 240 minutes and the absorption of the supernatant of MCR400 was measured at 550 nm. The change in the absorption of the supernatant of MCR400 was measured over time and heating temperature. The longer the heating time of the MCR400 suspension or the higher the heating temperature of the MCR400 suspension, the greater the absorption of the supernatant. At 20°C, the absorption of the supernatant of MCR400 increased from 0 to about 0.004 in 240 minutes. At 60°C, the absorption of the supernatant increased from about 0.01 to about 0.028 in 240 minutes. At 90°C, the absorption of the supernatant increased from about 0.022 to about 0.044 in 240 minutes. When MCR400 was heated at different temperatures for 240 min, the concentration of rhodamine B in the supernatant of the MCR400 suspension increased from 20 nM at 20 °C to 560 nM at 90 °C.

[0284] We can conclude from this example:

[0285] (i) It is possible to establish a relationship between changes in the luminous intensity of the MCR 400 and changes in the temperature of the MCR 400 and thus use the MCR 400 as a temperature probe.

[0286] (ii) The luminescence intensity of MCR400 increases with increasing temperature, a behavior opposite to that of free Rhodamine B in solution. This makes it possible in particular to distinguish the presence of free Rhodamine B from the presence of Rhodamine B associated with magnetosomes.

[0287] (iii) The luminescence intensity of MCR400 increases with increasing temperature, which can be attributed to the dissociation of rhodamine B from MCR400 when MCR400 is heated.

[0288] (iv) The amount of rhodamine B dissociated from the magnetosomes by heating can be evaluated by absorption measurement.

[0289] Example 4: Using MCR400 as a probe to detect irradiation

[0290] Three suspensions were maintained at a temperature of pH 7 and 25° C. The three suspensions contained 400 μg / mL of MCR400, 125 μM of Rhodamine B, or 400 μg / mL of ferric oxide MC mixed with 125 μM of Rhodamine B. These suspensions were irradiated with faxitron irradiation dosimeter (160 kV, 6.3 mA, without filter, 67.5 Gy / min). The irradiation dose was 0 Gy to 1350 Gy. When these suspensions were irradiated, the luminescence measured at 550 nm for the suspension containing Rhodamine B alone or the supernatant of the Rhodamine B mixed with MC was not changed or reduced. In contrast, when the MCR400 suspension was irradiated, the luminescence of the supernatant of the suspension, excited at 550 nm and measured at 576 nm, increased dramatically from 250 a.u. in the absence of irradiation to 600 a.u.-950 a.u. in the presence of irradiation greater than 250 Gy. These results indicate that Rhodamine B dissociates from MCR400 under the effect of irradiation. Furthermore, the emission wavelength of free Rhodamine B in the supernatant of MCR400 and in solution remained unchanged at 576 nm, regardless of the irradiation dose received by MCR400 or Rhodamine B. This indicates that free Rhodamine B in solution or Rhodamine B associated with magnetosomes was not altered by irradiation.

[0291] We can conclude from this example:

[0292] (i) It is possible to establish a relationship between changes in the luminescence of the MCR400 and changes in irradiance and thus use the MCR400 as an irradiance probe.

[0293] (ii) The luminescence intensity of the supernatant of the MCR400 suspension increases with increasing irradiation, a behavior that is different from that of free Rhodamine B in solution. This makes it possible in particular to distinguish the presence of free Rhodamine B from the presence of Rhodamine B associated with magnetosomes.

[0294] (iii) The enhanced luminescence of the MCR400 supernatant under irradiation can be attributed to the dissociation of Rhodamine B from MCR400 under irradiation.

[0295] (iv) The amount of rhodamine B dissociated from the magnetosomes by irradiation can be estimated by luminescence measurement.

[0296] Example 5: Using BNF-DiI as a temperature probe.

[0297] A suspension of DiI at a concentration of 5.4 μM and pH 7 was heated for 30 minutes at various temperatures between 20°C and 90°C using a dry heat bath. The maximum luminescence intensity of DiI excited at 530 nm increased between 20°C and 60°C, then decreased between 60°C and 100°C. When a suspension containing BNF-DiI mixed in water at a concentration of 80 μg / mL iron oxide was heated using a dry heat bath at temperatures between 20°C and 90°C, the behavior was different from that observed with DiI alone. In fact, the maximum luminescence intensity of BNF-DiI excited at 530 nm decreased between 20°C and 60°C, then remained almost stable between 60°C and 100°C.

[0298] In order to determine whether the luminescence change of BNF-DiI is due to the dissociation of DiI from BNF, the maximum luminescence intensity of the supernatant of BNF-DiI and DiI alone excited at 530nm was measured for suspensions containing BNF-DiI or DiI alone heated at different temperatures from 20°C to 90°C for a period of 0 to 30 minutes. When BNF-DiI was heated between 20°C and 90°C for a period of 0 to 30 minutes, the luminescence intensity of the supernatant of BNF-DiI did not increase. Given that the luminescence intensity of DiI did not significantly decrease when DiI was heated between 20°C and 90°C for a period of 0 to 30 minutes, this indicates that the concentration of DiI in the supernatant of the BNF-DiI suspension did not increase when the temperature was increased and DiI was therefore not dissociated from BNF.

[0299] We can conclude from this example:

[0300] (i) It is possible to establish a relationship between the change in the luminescence intensity of BNF-DiI and the temperature change of BNF-DiI and thus use BNF-DiI as a temperature probe.

[0301] (ii) The luminescence intensity of the BNF-DiI supernatant did not increase with increasing temperature, indicating that DiI did not dissociate from BNF due to the increase in temperature. In this case, the decrease in the luminescence of BNF-DiI with increasing temperature can be explained by the chemical modification of BNF-DiI.

[0302] Example 6: Using MCR400 as a pH probe.

[0303] MCR400 was tested as an in vivo pH probe on rat brain. For this purpose, 4 rats were used. Rat 1 was a healthy rat euthanized at the same time as rat 2. Rats 2 to 4 received 5 μl of 3×10 3Suspension of RG-2 cells, which were implanted at coordinates 2mm dorsal, 2mm lateral and 4mm depth using a stereotactic helmet. Rat 2 was not treated and euthanized 18 days after cell implantation. Rat 3 and rat 4 received 10μl of MCR400 suspension with a concentration of 668μg / ml iron oxide at the site of cell implantation for fourteen days after tumor cell implantation. Rat 3 and rat 4 were euthanized 2 hours and 4 days after the administration of magnetosomes, respectively. The brain was then extracted and cut into 3mm thick layers. A pulsed laser diode with a power of 40MHz, 1mW, and a wavelength of 405nm was then used to generate the light emission of the slices. The laser was coupled to an optical fiber with a diameter of 200μm positioned 1.5mm above the brain slice. The light emitted by the tissue was collected by a second optical fiber with a diameter of 365μm located at 600μm of the first optical fiber and positioned 1.5mm above the brain slice. The luminescence is detected by a spectrometer (B. Leh et al., J. Biomed. Opt. (2012) 17 (10), 108001), which enables the measurement of the luminescence intensity of the brain slice as a function of the wavelength of the emitted radiation. The two optical fibers can be moved laterally to different parts of the brain slice to identify the luminescence location.

[0304] For different rats, the luminescence spectrum of the healthy area without MCR400 was measured, and the luminescence spectrum was very similar, showing a luminescence band with the maximum intensity observed at about 500nm. This band is attributed to a small contribution of NADH luminescence at 450nm and the main contribution of flavoprotein at 525nm. At 575nm, a shoulder peak was observed, which will be the luminescence of the fat pigment also known as lipofuscin lipid fragments. At 625nm and 690nm, the luminescence of protoporphyrin IX and derivative molecules was observed. The spectrum of the healthy area is averaged, so that it is possible to normalize all the measured luminescence spectra of all slices of the rat brain.

[0305] The luminescence spectra of tumor areas without MCR400 were also measured for different rats and were very similar, showing the presence of peaks similar to those observed for healthy areas with lower intensities.

[0306] For rat 3, which was euthanized 2 hours after the administration of MCR400, the luminescence spectrum of the tumor site that received MCR400 showed the presence of two interesting peaks: the first at 569 nm, which was absent in the spectra of tumor areas and healthy areas without MCR400, and the second at 576 nm. We attribute these two peaks to the emission of MCR400 at 569 nm and the emission of rhodamine B and fat pigment at 576 nm. The peak at 576 nm is unlikely to come from the luminescence of fat pigment alone, because its relative luminescence intensity I 576 / I500 About 1, which is higher than the I measured in the luminescence spectra of healthy and tumor sites without MCR400. 576 / I 500 The relative luminous intensity is about 0.5, where I 576 and I 500 are the luminescence intensities measured at 576 nm and 500 nm, respectively.

[0307] For rat 4, which was euthanized four days after administration of MCR400, the luminescence spectrum of the tumor site that received MCR400 showed the presence of rhodamine B and lipochrome peaks at 576 nm. The MCR400 peak at 569 nm was no longer present. This can be explained by the dissociation of rhodamine B from MCR400 after administration of MCR400 in the rat brain.

[0308] In order to study whether this dissociation is attributed to the change of pH value, we first studied whether MCR400 is internalized in living cells, which can cause pH value changes.MCR400 and 500,000 MDA-MB-231 cells are placed in cell viability probe, i.e., in the presence of calcein-acetoxy-methyl ester (Ca), which is not a natural luminescent and luminescent in the presence of living cells.Cells are incubated for 6 hours in the presence of Ca or in the presence of Ca and MCR400, wherein the iron oxide concentration of MCR400 suspension is 31.5 μg / ml.Flow cytometer enables detection of the luminescence of Ca (FL1-H signal) or the luminescence of rhodamine B (FL3-H signal).FL1-H signal is weak (2-3) for single cells and increases to approximately 100 when cells are in the presence of Ca, which shows the vigor of MDA-MB-231 cells in the absence of MCR400. When the cells were incubated in the presence of Ca and MCR400, the FL1-H signal was greater than about 10 and most of the cells were alive. For the FL3-H signal, it showed that 99% of the cells were luminous in the presence of MCR400. In order to study MCR400 internalization, MCR400 with a concentration of 100 μg / ml iron oxide was placed in the presence of U87-Luc cells and macrophages for 2 hours at 37°C, and the cells were in their respective DMEM culture medium and RPMI culture medium containing 10% complement-free fetal bovine serum. U87-Luc cells were washed 5 times with PBS (phosphate buffered saline) and fixed, embedded in resin, and imaged by TEM (transmission electron microscopy). Macrophages were washed 5 times with PBS and then imaged by epifluorescence optical microscopy. In both cases, internalization of MCR400 in lysosomes was observed. During internalization of magnetosomes in lysosomes, the pH will acidify to values ​​typically between 3.5 and 5, which may induce the dissociation of Rhodamine B from the magnetosomes.

[0309] A suspension of MCR400 with 400 μg / mL iron oxide and a solution of 125 μM rhodamine B were mixed with hydrochloric acid solution or sodium hydroxide solution at concentrations in the range of 0.1 M to 12 M to adjust the pH value to a value between 2 and 12. Then, the sample containing the thus-treated MCR400 suspension was placed against a 0.6 T magnet at 4 °C for 12 hours and the supernatant of the treated MCR400 suspension was removed. The maximum luminescence intensity of the supernatant of the MCR400 suspension excited at 550 nm decreased from 140 a.u. at pH 2 to 20 a.u. at pH 12. On the other hand, the maximum luminescence intensity of rhodamine B alone excited at 550 nm increased from 440 a.u. to 580 a.u. This difference in behavior indicates that rhodamine B associated with MCR400 undergoes a transformation such as dissociation or chemical modification during the pH value change, which is different from rhodamine B. In addition, for pH values > 4, the emission wavelength of MCR400 is between 568 nm and 576 nm, which is lower than the emission wavelength of the supernatant of rhodamine B alone and MCR400 (for pH values > 4, the emission wavelength is between 577 nm and 580 nm). In contrast, for 2 < pH value < 4, the emission wavelengths of rhodamine B alone, MCR400, and the supernatant of MCR400 are similar, between 580 nm and 584 nm. These behaviors can be explained by the association of rhodamine B with magnetosomes at pH values > 4 and the dissociation of rhodamine B from magnetosomes at pH values < 4.

[0310] We can conclude that:

[0311] (i) It is possible to detect the luminescence change of MCR400 within a rat tumor. This luminescence change can be attributed to the pH value change induced by the internalization of MCR400 in cells, especially within organelles such as lysosomes or endosomes at acidic pH values, as we observed by electron transmission microscopy.

[0312] (ii) This luminescence change of MCR400 as a function of pH value can be attributed to the dissociation of rhodamine B from magnetosomes at acidic pH values.

[0313] Example 7: Using MCR400, BNF-Dil and MC-Dil as temperature and magnetic field probes.

[0314] Materials and methods:

[0315] For the experiment conducted on a mouse brain, MCR400 was introduced into the brain extracted from a mouse. In this case, 2 μl or 20 μl of an MCR400 suspension with a concentration of 20 mg / ml of maghemite was introduced at a depth of 1 mm in the brain.

[0316] For experiments performed in disrupted tissue, MCR400 magnetosomes were mixed with previously disrupted brain tissue. In this case, 2 μl or 20 μl of a 20 mg / ml MCR400 suspension was mixed with 2 mM 3 The brain tissue contained in the volume is mixed.

[0317] Then use laser diode to excite the fluorescence of the MCR400 suspension introduced into mouse brain or mixed with tissue at 405nm and detect by the fiber optic luminescence spectrometer described in Example 6.Fiber optic is positioned at the distance of 3mm above the surface of brain or broken tissue.This distance has been optimized to allow detection of the maximum fluorescence intensity emitted by MCR400.Under the situation that alternating magnetic field is not applied or frequency is 198kHz and intensity is the existence of the alternating magnetic field of 8mT or 25mT, measure the fluorescence of MCR400.Apply alternating magnetic field to produce the magnetic force excitation to MCR400.Also use Easir 2 thermal imaging infrared cameras and microprobe thermocouple (IT-18, Physitemp company (Physitemp, Clifton, USA) of Clifton, USA) to measure the temperature in the zone where MCR400 is located.

[0318] For the experiment carried out in water, three kinds of suspensions of MCR400, MC-DiI or BNF-Dil containing 3.6mg / mL maghemite concentration of 300 μ l are placed in tube and exposed to the application of the alternating magnetic field of intensity 25mT and frequency 198kHz, continue the different durations of 0 second, 100 second, 200 second, 300 second, 600 second, 900 second or 1200 second.For these suspensions mixed with their supernatant, the supernatant of these suspensions and these suspensions mixed in water, measure the luminescence of these suspensions.The suspension mixed in their supernatant is the suspension obtained after applying the alternating magnetic field.The supernatant of these suspensions is to obtain by placing 0.6T neodymium magnet against the quartz tube that holds these suspensions to keep different nanoparticles sticking to the tube wall.Then use pipette to suck supernatant.The suspension mixed in water is to obtain by removing supernatant from suspension and replacing it with water. The luminescence of the three sample types was excited at 405 nm (for MCR400) and 550 nm (for BNF-DiI and MC-DiI).

[0319] Results and discussion:

[0320] When 2 μl (40 μg) or 20 μl (400 μg) of MCR400 suspension was introduced into the mouse brain without applying a magnetic field, we have confirmed that neither the luminescence intensity nor the temperature of MCR400 changed over time. Similar results were obtained when MCR400 was exposed to the application of an alternating magnetic field with a frequency of 198 kHz and an intensity of 8 mT.

[0321] On the other hand, when 2 μl of the MCR400 suspension was introduced into the mouse brain and MCR400 was exposed to the application of an alternating magnetic field of frequency 198 kHz and intensity 25 mT for the first time (lane 1), the luminescence intensity and temperature of MCR400 increased from 5 arbitrary units (au) to 30 a.u. (i.e., an increase of 80%) and from 13° C. to 14° C., respectively, during the first 100 seconds of the application of the magnetic field. After the first magnetic treatment (lane 1) of 30 minutes, MCR400 was exposed to 4 other continuous magnetic excitations (lanes 2 to 5). For lanes 2 to 5, the luminescence changes were always present, but less obvious than in lane 1. No temperature changes were observed.

[0322] When 20 μ l of MCR400 suspension is introduced into the mouse brain and MCR400 is exposed to the application of the alternating magnetic field of frequency 198kHz and average intensity 25mT for 30 minutes, the luminous intensity and temperature increase. During the first 100 seconds of magnetic force excitation, for the applied MCR400 of 20 μ l, the luminous intensity increased from 20 a.u. to 90 a.u., i.e. increased by about 80%. For the applied MCR400 of 2 μ l and 20 μ l, the luminous increase percentage is similar, i.e. about 80%. During the first 100 seconds of magnetic force excitation, the temperature of MCR400 increased from 9 ℃ to 12 ℃. The temperature increase percentage of about 33% is significantly lower than the luminous increase percentage.

[0323] When 2 μl of MCR400 suspension was mixed with brain tissue and MCR400 was exposed to the application of an alternating magnetic field with a frequency of 198 kHz and an intensity of 25 mT for the first time within 30 minutes, the luminescence and temperature of MCR400 increased by 30 times and 2°C, respectively, during the first 100 seconds of magnetic excitation, both increases being greater than those observed in the brain.

[0324] When 300 μl of suspensions of MCR-400, BNF-Dil and MC-Dil containing 3.6 mg / ml maghemite were exposed to the application of a magnetic field with an intensity of 25 mT and a frequency of 198 kHz for different times of 0, 100, 200, 300, 600, 900 and 1200 s, a strong increase in the luminescence of the supernatant was observed during the first 100 s of magnetic excitation, with the supernatant of MCR400 increasing from 1 a.u. to 1 9a.u., the supernatant of BNF-Dil increased from 0a.u. to 0.32au and the supernatant of MC-Dil increased from 0a.u. to 0.2au, and then after 100 seconds, the luminescence of the supernatant of MCR-400 was relatively stable at 18.5au-19.5au, the luminescence of the supernatant of BNF-Dil was relatively stable at 0.31au-0.33au and the luminescence of the supernatant of MC-Dil was relatively stable at 0.17au-0.2au. In contrast to the luminescence of the supernatant without particles, when an alternating magnetic field with an intensity of 25 mT and a frequency of 198 kHz was applied for different durations of 0 s, 100 s, 200 s, 300 s, 600 s, 900 s and 1200 s, the luminescence of MCR-400, BNF-Dil and MC-Dil mixed with their supernatants or water remained stable at a low value of about 4 for MCR400 or remained at zero for BNF-Dil and MC-Dil.

[0325] We also measured the slope ΔF / δt at the starting point of the luminescence change of MCR400 introduced into the mouse brain or mixed with tissue as a function of the slope ΔT / δt at the starting point of the temperature change. As ΔT / δt increases, ΔF / δt increases and a relationship close to a linear relationship can be established between ΔF / δt and ΔT / δt, where the coefficient (ΔF / δt) / (ΔT / δt) is about 45 a.u. / °C. In addition, when ΔT / δt is about 0, a non-zero fluorescence change can be detected, and ΔF / δt is about 0.03.

[0326] We can conclude from this example:

[0327] (i) When the intensity of the alternating magnetic field is below a certain threshold (B<8mT), it produces neither luminescence change nor temperature change of MCR400. Magnetic excitation of MCR400 that causes changes in luminescence and / or temperature requires the application of an alternating magnetic field with an intensity greater than 8mT.

[0328] (ii) When MCR400 is introduced into the brain or mixed with tissue and exposed to an alternating magnetic field of an intensity greater than about 25 mT, this can induce an increase in luminescence and a rise in temperature. The change in luminescence is more obvious than the change in temperature. In addition, the luminescence change can be detected under conditions where temperature changes cannot be detected, particularly at low concentrations of iron oxide nanoparticles. This makes this luminescent probe a more sensitive local probe than a macroscopic temperature probe that measures average temperature on a larger volume.

[0329] (iii) The increase in luminescence of MCR400 induced by the application of an alternating magnetic field can be related to the dissociation of rhodamine B from the magnetosomes. The luminescence is initially quenched when rhodamine B associates with iron oxide nanoparticles and increases when rhodamine B dissociates from MCR400 after the application of an alternating magnetic field. Rhodamine B then diffuses into the probe environment, which can explain the occasional random changes in luminescence intensity observed after 100 seconds.

[0330] (iv) The increase in luminescence of the supernatant of MCR400 can be attributed to the temperature increase caused by the magnetic field. In contrast, the increase in luminescence of the supernatant of BNF-DiI is not due to the temperature increase induced by the magnetic field, but to the non-thermal effect induced by the magnetic field.

[0331] (v) After exposing MCR400 to an alternating magnetic field of 25 mT and 198 kHz twice for more than 30 minutes, the luminescence increase of MCR400 produced by the application of the alternating magnetic field decreased. This quenching can be attributed to the diffusion of MCR400 under the alternating magnetic field or the reduction in the number of Rhodamine B molecules associated with the magnetosomes.

[0332] (vi) The luminescence change and temperature change of MCR400 induced by applying an alternating magnetic field of intensity 25mT and frequency 198kHz are related. The greater the increase in luminescence change, the greater the increase in temperature change. In addition, a quasi-linear relationship between ΔF / δt and ΔT / δt can be established for a series of different conditions: (i) for different amounts of iron oxide nanoparticles tested (40μg or 400μg); (ii) for different types of nanoparticles (magnetosomes and BNF); (iii) for different types of fluorophores (rhodamine B and Dil) and for different types of preparations (MCR400 mixed with tissue, inserted into the brain or mixed in water).

[0333] (vii) In order to establish the relationship between the luminescence variation and the temperature variation, it is possible to consider the variation that occurs during the first 100 to 200 seconds of excitation due to the alternating magnetic field. In fact, after this initial period of 100 to 200 seconds, the luminescence variation of the MCR400 may be random.

[0334] (viii) When the suspension of MCR400, MC-Dil, BNF-Dil was exposed to the application of an alternating magnetic field of intensity 25 mT and frequency 198 kHz, this produced an increase in the luminescence intensity of the supernatant observed during the first 100 seconds of magnetic excitation. This luminescence increase can be explained by the dissociation of Rhodamine B or Dil from the magnetosomes or BNF. When Rhodamine B or Dil is free in the solution and the iron oxide nanoparticles have been removed, their luminescence will increase because it will no longer be quenched by the iron oxide nanoparticles.

[0335] (ix) After applying an alternating magnetic field of 25 mT and 198 kHz, the luminescence of MCR400, BNF-Dil and MC-Dil mixed with supernatant or water did not change. This behavior can be explained by the absorption of light by iron oxide nanoparticles.

[0336] Example 8: Measurement of the lifetime of Rhodamine B and MCR400.

[0337] The life experiment was carried out using the frequency method. The configuration used a modulated laser diode as an excitation source, a confocal fluorescence spectroscope, a control unit and camera for enhancing the image, a frequency generator, and a spectrometer controller. The modulation frequency varied between 30MHz and 180MHz. The excitation wavelength was 445nm. The laser was modulated by a radio frequency signal. An interference filter suppressed parasitic fluorescence from the laser diode and a high-pass filter was placed in the detection path to reduce scattering. A suspension of 1mL volume of 5mg / mL iron oxide MCR400 and 5μM concentration of rhodamine B was placed in a dry heat bath for 5 minutes to heat the suspension at 25°C and 45°C. 10μl of a suspension of 5mg / ml iron oxide MCR400 and 5μM concentration of rhodamine B was deposited on a slide and then placed in front of a ×40 objective to measure the fluorescence lifetime of each suspension.

[0338] At 25° C., the fluorescence lifetime of MCR400 is 4.8 ns and the fluorescence lifetime of Rhodamine B is 1.6 ns. When the sample is heated to 45° C., the lifetime of MCR400 is 1.6 ns and the lifetime of Rhodamine B is 1.3 ns.

[0339] We can conclude from this example:

[0340] (i) The fluorescence lifetime of Rhodamine B is shorter than that of MCR400 at 25°C and 45°C.

[0341] (ii) Rhodamine B is well associated with magnetosomes in MCR400 because the fluorescence lifetime of Rhodamine B is shorter than that of MCR400.

[0342] (iii) When the temperature increases, the lifetime of MCR400 decreases more significantly than that of Rhodamine B.

[0343] (iv) At 45°C, the lifetime of MCR400 is the same as that of Rhodamine B alone, indicating that Rhodamine B dissociates from the magnetosomes.

[0344] Example 9: Use of NANOF in association with a drug compound Uses

[0345] When a drug compound is mixed with NANOF, the compound associates with NANOF by a chemical reaction. The particles are then delivered or inserted into an organism or a part of an organism, such as a tumor or tumor cell. The particles are then exposed to a magnetic field, such as an alternating magnetic field, or to changes in the pH value, temperature or chemical composition of the medium surrounding the particles, which induces the dissociation of the drug compound from NANOF and / or chemical modification of NANOF. After this dissociation, the compound is activated and used as a diagnostic or therapeutic treatment tool.

[0346] Example 10: Non-pyrogenic and luminescent proteins synthesized by magnetotactic bacteria coated with poly-l-lysine and Dil Synthesis of NANOF(M-PLL-DiI)

[0347] The pyrogen-free and luminescent NANOF suspension of M-PLL-DiI was prepared under pyrogen-free conditions under a fume hood using the central portion of the MSR-1 magnetosomes prepared as follows. The MSR-1 bacteria were first grown on an agar gel at 30°C in the presence of iron and under low oxygen concentration (about 0.5% O 2 ) for 5 to 7 days. The magnetic colonies were removed and cultured in an iron-free preculture medium containing a carbon source, a nitrogen source, a mineral source, a trace element source and a yeast extract source in the presence of air at 30°C for several days. The magnetotactic bacteria collected from the preculture were cultured in a 50-liter fermentor at 30°C in a medium similar to the preculture medium. During the growth period, the pH was maintained at 6.8-7 by adding an acidic nutrient medium containing an iron source, and compressed air was introduced into the culture medium to promote bacterial growth while maintaining the oxygen concentration below 0.2% to allow the synthesis of magnetosomes. The MSR-1 bacteria derived from the fermentation were concentrated to an optical density (OD) of 110-120 measured at 565nm. 565). 100 ml of the bacterial concentrate was then mixed with 400 ml of 5M NaOH and heated at 60° C. for 1.5 to 2 hours to lyse the bacteria. The treated magnetosomes were then separated from the bacterial fragments by placing a neodymium magnet against the wall of the container containing the lysed bacterial suspension overnight and replacing the supernatant containing NaOH and bacterial fragments with 1×PBS. The resulting suspension was then sonicated at 10 W for 20 seconds in the presence of 1×PBS, placed against a neodymium magnet for 15 minutes, the supernatant was removed and the treated magnetosomes were resuspended in 1×PBS. The sequence of sonication and magnetic separation was repeated 4 times. 100 mL of the resulting suspension was then mixed with 200 mL of a solution containing 1% Triton X-100 and 1% SDS, the mixture was heated at 50° C. overnight, placed against a neodymium magnet, the supernatant was removed and replaced with 80 mL of phenol (pH 8). The resulting suspension was heated for 2 hours by ultrasonic treatment at 60°C, maintained overnight at 60°C without ultrasonic treatment, placed against a magnet, the supernatant of the suspension removed and replaced with 80 mL of chloroform. The suspension containing chloroform was placed against a neodymium magnet, the supernatant removed and the residual chloroform adsorbed on the surface of the treated magnetosomes removed by heating the magnetosomes (without supernatant) for 2 hours in a fume hood. Finally, the central portion of the obtained magnetosomes was desorbed from the glass wall of the tube containing them by adding 80 ml of 1M NaOH heated at 60°C for 1 hour in an ultrasonic bath. The suspension containing the central portion of the magnetosomes was placed against a neodymium magnet, the supernatant removed and replaced with sterile MilliQ water, and the suspension was ultrasonically treated for 20 seconds at 10 W. This washing sequence was repeated 4 times. The suspension containing the central portion of the magnetosomes was degassed with nitrogen to avoid oxidation, sterilized by autoclaving and stored at -80°C.

[0348] The suspension containing the central part of the magnetosomes was characterized by an endotoxin concentration of 10 to 100 EU / mg / ml; low stability and rapid sedimentation; and the absence of a coating as observed by transmission electron microscopy.

[0349] A 10 ml suspension containing 56 mg of the central part of the magnetosome was mixed with 373 mg of poly-L-lysine hydrobromide with a molar mass of 21000 g / mol (Gmac, catalog number: 25988-63-0) and 250 μg of Dil. The mixture was sonicated at 25 kHz in an ultrasonic bath at 30°C for 15 minutes. The pyrogen-free mixture was then stirred at 4°C on a rotating wheel at a speed of 13 rpm for 19 hours. Then, in the dark and at 5°C, the mixture was sonicated at 10W for 10 minutes with a 1 minute pulse and a time interval of 1 minute between pulses. The suspension was concentrated by centrifugation at 14,000 rpm at 5°C for 30 minutes, then washed twice with 0.5% ethanol and washed with sterile The suspension thus obtained was finally resuspended in sterile water to obtain a pyrogen-free suspension of M-PLL-DiI.

[0350] 3T3 cells were cultured in DMEM containing 10% NCBS at a density of 5 × 10 cells / 2 ml. 5 The concentration of cells was inoculated into a Petri dish. After 24 hours, they were rinsed with PBS and then treated with a suspension of M-PLL-DiI (250 μg / ml (in terms of iron)) for 24 hours. After treatment, the cells were rinsed three times with a medium containing 10% NCBS and then photographed using an epifluorescence microscope. Cellular internalization of MPLL-DiI was observed by fluorescence.

[0351] Example 11: Characteristics of MCR400 compared with those of MC

[0352] Electron transmission microscopy measurements revealed that MCR400 had an average size of 60 nm and a size distribution of 15 nm to 90 nm, while MC had an average size of 40 nm and a size distribution of 5 nm to 60 nm. Since the magnetosome size of MCR400 is increased by 50% compared to MC, we can expect that the coercive force, the ratio between the residual magnetization and the saturation magnetization, or SAR, of MCR400 will increase by at least 0.01%, 0.1%, 1%, 2%, 5%, 10%, 25%, or 50% compared to MC.

Claims

1. A method for detecting a change in luminescence, comprising placing a particle probe comprising a particle at a location of interest and detecting a change in luminescence of the particle probe, wherein the particle comprises: at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle having a non-zero coercivity associated with at least one compound by weak bonds, wherein the weak bond allows dissociation of the compound from the nanoparticle upon physicochemical perturbation, in, The compound is a luminescent substance having at least one chemical functional group selected from the group consisting of carboxylic acids, esters, thiols, amines, ethers, and acidic, basic, oxidized, reduced, neutral and positively or negatively charged derivatives of these functions, wherein the nanoparticles comprise a crystalline core and a coating of non-biological origin, Wherein, the luminescent substance is associated with the nanoparticle in at least one of the following ways: (i) the luminescent substance is in direct contact with the nanoparticles, so that there is no substance between the luminescent substance and the nanoparticles, (ii) the luminescent substance is incorporated into or adsorbed onto the crystalline core of the nanoparticle, and (iii) the luminescent substance is incorporated into the coating of the nanoparticle or adsorbed thereon, wherein the association is such that the luminescence intensity of the luminescent substance associated with the ferrimagnetic or ferromagnetic iron oxide nanoparticles is quenched by the ferrimagnetic or ferromagnetic iron oxide nanoparticles compared to the luminescence intensity of the luminescent substance alone, wherein, when the luminescent substance is separated from the nanoparticle and ends up in the environment of the nanoparticle, the luminescent substance dissociates from the nanoparticle, wherein the dissociation results in an increase in the luminescence intensity of the luminescent substance, and Optionally, the environment of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle is a liquid medium, a solid medium or a gaseous medium or at least one substance different from the compound, which surrounds or includes the particle at a distance greater than 10 nm and less than 1 m from the center or outer surface of the particle, wherein the dissociation of the luminescent substance from the iron oxide nanoparticles results in or causes: i) a dissociation or modification associated with a localized luminescence change that occurs when the luminescent species diffuses at a distance measured from the center of the ferrimagnetic or ferromagnetic iron oxide nanoparticle, and ii) The luminescent substance is located in a liquid medium, a solid medium or a gaseous medium, which surrounds or includes the particle at a distance of 10 nm to 1 m measured from the center or the outer surface of the particle.

2. The method according to claim 1, wherein the luminescence, luminescence variation and luminescence characteristics are defined as follows: - luminescence is the light-emitting property of said luminescent substance, - the luminescence change is a change in the luminescence properties of the luminescent substance, - the luminescence property is selected from the group consisting of: luminescence wavelength, luminescence intensity, luminescence lifetime, and luminescence anisotropy of the luminescent substance.

3. The method of claim 1, wherein the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle is synthesized by a living organism.

4. The method of claim 1, wherein said dissociation of said compound does not result in: partial or total destruction of said compound, denaturation of the compound, The compound loses at least 10 10 atoms, said compound loses at least 90% of its weight, The luminescence intensity of the compound is reduced by more than 90%, or The compound loses its therapeutic or diagnostic properties.

5. The method according to claim 1, wherein the luminescent substance is a substance selected from the group consisting of: i) a substance which is capable of luminescence when it is or has been exposed to optical radiation, ii) fluorescent substances, iii) substances having luminescent or luminescent properties, iv) luminophore, v) chromophore, and vi) Fluorophores, The luminescence or luminescence characteristic is optionally selected from the group consisting of: luminescence wavelength, luminescence intensity, luminescence lifetime, and luminescence anisotropy.

6. The method of claim 1, further comprising chemical modification of the compound, wherein the chemical modification is a compositional and / or structural change that induces a change in at least one property of the at least one compound.

7. The method of claim 1, wherein the particles do not contain the following: crystalline liquids, dendritic polymers, greater than 90% by mass of carbon materials or organic materials derived from living organisms, or greater than 10 3 Endotoxin units.

8. The method of claim 1, wherein the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle has at least one property selected from the group consisting of: a crystalline core of iron oxide, wherein the crystallinity of the crystalline core is characterized by at least one crystal plane, A coating of non-biological origin, wherein the coating provides at least one property selected from the group consisting of: enabling the transfer of energy or particles between the compound and the at least one ferrimagnetic or ferrimagnetic iron oxide nanoparticle, stabilizing the at least one ferrimagnetic or ferrimagnetic iron oxide nanoparticle, rendering the at least one ferrimagnetic or ferrimagnetic iron oxide nanoparticle biocompatible, promoting cellular internalization of the at least one ferrimagnetic or ferrimagnetic iron oxide nanoparticle, and coupling the at least one ferrimagnetic or ferrimagnetic iron oxide nanoparticle to a different substance, Ferrimagnetic or ferromagnetic behavior, Single magnetic domain, Chain arrangement, Measured in sizes ranging from 1 nm to 1 μm in the presence or absence of a coating, an electric charge, which optionally varies with pH in the presence or absence of a coating and is capable of promoting cellular internalization, The at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle comprises at least one iron atom and at least one oxygen atom, The at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle comprises at least one iron atom and at least one oxygen atom, the magnetic moments of which are antiparallel, thereby providing ferrimagnetic properties, The at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle comprises at least one iron atom and at least one oxygen atom whose magnetic moments are parallel, thereby providing ferromagnetic properties, and Said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle is responsible for the dissociation of said compound from said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle and / or the chemical modification of said compound after applying a physicochemical disturbance to said particle.

9. The method of claim 1, wherein the modification of the particles is selected from the group consisting of: The pH of the particles increases or decreases within 10 -3 and 10, The temperature of the particles increases or decreases within 10 -13 and 10 3 ℃, The charge of the particles increases or decreases within 10 -3 and 10 2 The range between volts, The number of atoms contained in the particles increases or decreases between 1 and 10 10 between atoms.

10. The method of claim 1, wherein the physicochemical interference has at least one effect selected from the group consisting of: i) inducing a greater change to said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle than to said compound, ii) inducing a change in said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle without inducing such a change in said compound, iii) reversibly changing at least one property of said compound or said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle, iv) causing a temperature increase of less than 300°C to the particles, and v) The particles suffer a mass loss of less than 90%.

11. The method of claim 1, wherein the physicochemical interference is, is caused by, or induces at least one effect selected from the group consisting of: i) a change in the environment of the particle, wherein the environment of the particle is a liquid medium, a solid medium or a gaseous medium, or at least one substance surrounding or including the particle, ii) a change in the environment of the particle selected from the group consisting of: a change in pH of the environment within 10 -3 and 10 pH units, the temperature of the environment changes by 10 -13 and 10 3 ℃, the redox potential of the environment changes between 0.001 and 100 V, and the viscosity of the environment changes between 10 -9 and 10 20 The concentration of at least one substance in the environment varies between 10 Pa.s and -13 and 10 10 mol / L, micromol / L, nanomol / L, mol / mL, micromol / mL, nanomol / mL, mol / m3, mol / dm3, mol / cm3, or mol / mm3, iii) a change in at least one state of the particles selected from the group consisting of: a change in pH of the particles within 10 -3 and 10 pH units, and a temperature change of 10 -13 and 10 3 ℃, the standard potential changes between 0.001V and 100V, the charge of the particles increases or decreases between 0.001 and 100 volts, and the number of atoms contained in the particles changes between 1 and 10 10 Between atoms, iv) the concentration of at least one substance in the environment of the particle changes by more than 10 -13 mol / L, micromol / L, nanomol / L, mol / ml, micromol / ml, nanomol / ml, mol / m3, mol / dm3, mol / cm3, or mol / mm3, v) the chemical composition of at least one substance in the environment of the particle changes by less than 10 10 mol / L, micromol / L, nanomol / L, mol / ml, micromol / ml, nanomol / ml, mol / m3, mol / dm3, mol / cm3, or mol / mm3, vi) a change in at least one substance in the environment of the particle selected from the group consisting of a chemical change, a structural change, the appearance of at least one substance in the environment, the disappearance of at least one substance in the environment, and combinations thereof, and vii) the environment of the particles is less than 10 10 a change in the chemical composition of a substance in the environment, wherein the change is selected from the group consisting of: a chemical change, a structural change, the appearance of at least one substance in the environment, or the disappearance of at least one substance in the environment, and Its combination.

12. The method of claim 1, wherein the particles undergo a transition from an inactivated state to an activated state under the physicochemical perturbation, wherein the transition is characterized by at least one property selected from the group consisting of: Constant K D In 10 -9 and 1, the constant K D is equal to the ratio of the concentration of said compound associated with said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle measured when said particle is activated divided by the concentration of said compound associated with said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle measured when said particle is deactivated, The particles have a weight loss of less than 90%, and The luminescence intensity I of the compound connected to the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle L less than the luminescence intensity I of the compound not attached to the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle NL .

13. The method according to claim 1, wherein the surface / volume ratio of the particles is between 10 -10 and 10 3 nm -1 between.

14. The method of claim 1, wherein the dissociation of the luminescent substance from the iron oxide nanoparticles has or results in at least one property selected from the group consisting of: (i) reversible dissociation involving the dissociation of the luminescent substance from the ferrimagnetic or ferromagnetic iron oxide nanoparticles, followed by a return to the initial state of the particles obtained when the particles were deactivated, (ii) global dissociation, involving the entire luminescent substance, (iii) dissociation or modification associated with the movement, translation, oscillation, or vibration of the luminescent substance or the ferrimagnetic or ferromagnetic iron oxide nanoparticles, (iv) dissociation or modification associated with the coupling between the electromagnetic field, the luminescent substance and the ferrimagnetic or ferromagnetic iron oxide nanoparticles, (v) dissociation or modification due to changes in the environment of the particle, and (vi) This is followed by dissociation or modification of the luminescent substance or a portion thereof which diffuses into the particle environment.

15. The method of claim 1, wherein the dissociation of the luminescent substance from the ferrimagnetic or ferromagnetic iron oxide nanoparticles results in the luminescence change having at least one characteristic selected from the group consisting of: (i) irreversible dissociation, involving the dissociation of the luminescent substance from the ferrimagnetic or ferromagnetic iron oxide nanoparticles without the luminescent substance returning to its initial state, and (ii) Partial, involving the dissociation or modification of a part, chemical group or atom of the luminescent substance.

16. The method of claim 1, wherein the luminescent substance has or causes at least one property selected from the group consisting of: i) transferring energy or particles to the ferromagnetic or ferrimagnetic iron oxide nanoparticles, or receiving energy or particles from the ferromagnetic or ferrimagnetic iron oxide nanoparticles, ii) the separation distance between the luminescent substance and the ferromagnetic or ferrimagnetic iron oxide nanoparticles is less than 1 cm or less than 100 times the Förster distance, iii) the characteristics of the donor or recipient in the terms used to describe Forster transfer, iv) the ability to emit light at one or more wavelengths at which the ferromagnetic or ferrimagnetic iron oxide nanoparticles absorb light, (v) a lifetime or luminescence intensity measured when the luminescent substance is alone, which is different from the lifetime or luminescence intensity of the luminescent substance combined with the ferromagnetic or ferrimagnetic iron oxide nanoparticles, so that the change in the lifetime or luminescence intensity is measurable; and (vi) the lifetime of the light-emitting substance alone is greater than 0.5 ns, and Wherein the at least one property causes a change in at least one luminescence property of the luminescent material associated with the ferrimagnetic or ferromagnetic iron oxide nanoparticles exposed to the physicochemical disturbance, which change is different from the change in the same at least one luminescence property of the luminescent material alone exposed to the same physicochemical disturbance.

17. The method of claim 1, wherein the iron oxide nanoparticles have or cause at least one property selected from the group consisting of: i) a non-zero absorption, absorption coefficient, absorption index, or extinction coefficient at the emission wavelength of the luminescent substance, ii) a material contained in its core and / or coating that absorbs at the emission wavelength of said luminescent substance, iii) a size larger than that of superparamagnetic iron oxide nanoparticles or a size larger than 0.1 nm, iv) ferrimagnetic or ferromagnetic properties, which increase the absorption, absorption coefficient, absorption index, or extinction coefficient of the ferrimagnetic or ferromagnetic iron oxide nanoparticles and / or promote the coupling between the dipole and / or magnetic moment of the iron oxide nanoparticles and the dipole and / or magnetic moment of the luminescent substance, and v) crystallinity, which enables energy and particle transfer between the ferrimagnetic or ferromagnetic iron oxide nanoparticles and the luminescent substance, and wherein the at least one property results in a change in at least one luminescence property of the luminescent material associated with the iron oxide nanoparticles exposed to the physicochemical disturbance that is different from the change in the same at least one luminescence property of the luminescent material alone exposed to the same physicochemical disturbance.

18. The method of claim 1, wherein the luminescent substance dissociates from the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle, the physicochemical disturbance induces dissociation of weak bonds between the luminescent substance and the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle, and the luminescent substance has at least one luminescent property that changes during the dissociation between the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle and the luminescent substance.

19. The method of claim 1, wherein the environment of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle is a liquid medium, a solid medium or a gas medium or at least one substance different from the luminescent substance, which surrounds or includes the particle at a distance of 10 nm to 1 m measured from the center or outer surface of the particle.

20. The method of claim 1, wherein the physicochemical disturbance is a disturbance caused by radiation and / or changes in the environment of the particles.

21. The method of claim 20, wherein the radiation is wave radiation or particle radiation, sound wave radiation, ionizing radiation, X-ray radiation, electromagnetic radiation, radiation due to electric, magnetic or electromagnetic fields, or radiation caused by neutrons, protons, electrons, positrons, alpha particles, beta, gamma, neutrinos or muons.

22. The method of claim 20, wherein the change in the particle's environment is a change in pH, temperature, redox potential, viscosity or chemical composition of the environment.

23. The method of claim 1, wherein the compound and the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle are separated by a K D Weak bonds, or hydrogen bonds, bonds due to van der Waals, London, or Debye forces, intermolecular or inter-nanoparticle forces, or hydrophobic, hydrophilic, or electrostatic interactions, Where K D is equal to the ratio of the concentration of said compound associated with said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle measured when said particle is activated divided by the concentration of said compound associated with said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle measured when said particle is deactivated, wherein the particles are inactivated when no physicochemical disturbance is applied to the particles, wherein the particles are activated when a physicochemical disturbance is applied to the particles, wherein optionally when said compound is in direct or indirect contact with said ferrimagnetic or ferromagnetic iron oxide nanoparticle, said compound binds to said at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle, wherein optionally when the distance between the compound and the ferrimagnetic or ferromagnetic iron oxide nanoparticles is less than 10 μm, 1 μm or 100 nm, the compound is in direct contact with the ferrimagnetic or ferromagnetic iron oxide nanoparticles, Wherein optionally at least one substance, polymer or solvent is located between the ferrimagnetic or ferromagnetic iron oxide nanoparticles and the compound, the compound is in indirect contact with the ferrimagnetic or ferromagnetic iron oxide nanoparticles.

24. The method of claim 1, wherein the compound and the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle are bound by a weak bond, and the weak bond is not a strong bond, wherein the strong bond is a covalent bond or an ionic bond, or a bond that prevents the compound from dissociating from the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle.

25. The method of claim 1, wherein the luminescent substance is selected from the group consisting of: lissamine rhodamine B LDS 751, rhodamine red-x, rox, red 613, rhodamine, rhodamine B, TRITC X-rhodamine.

26. A method according to claim 18, wherein the luminescent material has at least one luminescent property, and the luminescent property changes under the operating conditions of the particles, wherein the operating conditions of the particles correspond to the following conditions: the particles are transformed from an inactivated state in which no physicochemical disturbance is applied to the particles to an activated state in which a physicochemical disturbance is applied to the particles, or vice versa from an activated state to an inactivated state.

27. The method of claim 1, wherein the particle is a drug, a therapeutic substance, a contrast agent, or a diagnostic agent.

28. The method of claim 18, wherein the luminescence change is: i) an increase or decrease in at least one luminescence property of the particles or luminescent substances, ii) Continuous increase in luminescence properties, over a period longer than 10 -10 seconds, a continuous increase of at least 1 and a decrease of at least 1 occurring during the random interaction of the luminescent material, iii) characterized by the slope ΔL / δt at the starting point, where δt represents the duration originating from the onset of the luminescence change, which is determined during the first seconds or minutes after the physicochemical perturbation applied to the particles, iv) determined in an area that enables detection of a change in luminescence or an area containing a sufficient number of particles or an area containing more than 1 particle per unit volume, v) due to: (a) a change in the transfer of energy or particles between the luminescent substance and the ferrimagnetic or ferromagnetic iron oxide nanoparticles; (b) a change in the distance separating the luminescent substance from the ferrimagnetic or ferromagnetic iron oxide nanoparticles; or (c) a change in the chemical composition or structure of the luminescent substance, vi) occurs when the particles interact with synthetic or non-synthetic substances, and / or vii) causing a physicochemical interference induction of greater than 10 on the luminescent substance associated with the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle -10 % change in luminescence, whereas the same perturbation induces less than 90% change in luminescence when the same perturbation is applied to the luminescent substance alone that is not associated with the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle.

29. The method of claim 28, wherein the change in the transfer of energy or particles between the luminescent substance and the ferrimagnetic or ferromagnetic iron oxide nanoparticles is starting, increasing, stopping or decreasing.

30. The method of claim 8, wherein the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle has ferrimagnetic or ferromagnetic behavior characterized by at least one of the following properties: i) Coercivity greater than 1Oe; ii) a ratio between the residual magnetization and the saturation magnetization greater than 0.01, and iii) a saturation magnetization greater than 0.1 emu per gram of said ferrimagnetic or ferromagnetic iron oxide nanoparticles.

31. The method of claim 8, wherein the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle has ferrimagnetic or ferromagnetic behavior measured at a temperature above 0.1 K.

32. The method of claim 1, wherein the physicochemical perturbation applied to the particles does not significantly change at least one property of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle, wherein the significant change in the property of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle is selected from the group consisting of: i) the size of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle increases or decreases by more than 10 μm, ii) the size distribution of the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle increases or decreases by more than 10 μm, iii) the nanoparticles are transformed from single-domain magnetic nanoparticles into multi-domain magnetic nanoparticles, iv) converting nanoparticles consisting of iron oxide into nanoparticles consisting only of iron or oxygen, and v) Transformation of nanoparticles from crystalline state to amorphous state.

33. The method of claim 1, wherein the physicochemical disturbance is imparted to the particles by radiation.

34. The method of claim 33, wherein the radiation is selected from the group consisting of: i) wave radiation, ii) particle radiation, iii) acoustic radiation, iv) ionizing radiation, v) X-ray radiation, vi) electromagnetic radiation, vii) radiation due to electric, magnetic or electromagnetic fields, or radiation caused by neutrons, protons, electrons, positrons, alpha particles, beta, gamma, neutrinos or muons.

35. The method of claim 1, wherein dissociation of the luminescent species from the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle suppresses or reduces quenching of luminescence intensity of the luminescent species.

36. The method of claim 1, wherein the ferrimagnetic or ferromagnetic iron oxide nanoparticles have a size of 20 nm to 150 nm.

37. The method of claim 1, wherein the particles allow for energy and particle transfer between the ferrimagnetic or ferromagnetic iron oxide nanoparticles and the luminescent substance.

38. The method of claim 1, wherein the luminescent substance is in direct contact with the nanoparticle.

39. The method of claim 8, wherein the at least one ferrimagnetic or ferromagnetic iron oxide nanoparticle has a chain arrangement, wherein the chain has at least one property selected from the group consisting of: i) the chain comprises from 2 to 500 nanoparticles, ii) the nanoparticles belonging to the chain have a crystallographic direction, easy axis of magnetization, or field lines oriented in the direction of chain elongation, and iii) The size of the chains is less than 10 μm.

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