Patent Application
20240156745
The field of the invention is that of nanoparticles, comprising two centers of activity or free radical capture/production, for improving the sensitivity to radiation of the nanoparticles, and that of a pathological body part, for example a tumor, comprising nanoparticles or at least one constituent of a composition comprising nanoparticles preferentially with at least two centers of activity or free radical production in the core and coating of the nanoparticle or at least one constituent of the compositions, respectively, which are preferentially irradiated preferentially resulting in an improved treatment efficacy compared with standard irradiation conditions performed in the absence of such composition.
Some nanoparticles are known to be radiosensitive. Here, we propose to introduce two centers of production or capture of free radicals to increase the radiosensitivity of the nanoparticles, i.e. preferentially the production of free radical species is more important when nanoparticles comprising these two centers are exposed to radiation than when nanoparticles comprising only one center or no center are exposed to radiation.
Nanoparticles or at least one constituent of a composition can be associated with a photosensitizing molecule, for example Protoporphyrin IX was associated with a gold nanoparticle or at least one constituent of the composition to yield efficient photosensitizing effect in cervical cancer therapy, (H. Eshghi et al, Photodiagnosis and Photodynamic Therapy, V. 10, P. 304-312 (2013)). Here, we introduce the new feature of preferentially inserting at least two centers activity or of free radical production or radiation amplification, where preferentially one of these centers is inserted in the core of the nanoparticle or at least one constituent of the composition and the other center is inserted in the coating of the nanoparticle or at least one constituent of the composition. Such configuration preferentially presents the advantage of increasing the quantity of free radicals that is produced or of amplifying the radiation compared with a nanoparticle or at least one constituent of the composition comprising only one center of activity or free radical production or radiation amplification. The presence of two centers located in two different parts of the nanoparticle or at least one constituent of the composition preferentially enables applying the treatment through a spatial sequence, i.e. with free radical production or radiation amplification occurring at two different locations, where these two locations are preferentially separated by a nanometric distance or 1 or 5 or 10 nm, hence potentially enabling to reach a large concentration of these centers in the treated body part, hence potentially enabling to achieve a synergic effect of these two centers. The arrangement in chains of the nanoparticles or at least one constituent of the composition can also be a way to increase the number of centers activity or of free radical production or radiation amplification in the treated body part, given that the available surface into which the centers can be introduced is enhanced both by the presence of the nanoparticle or at least one constituent of the composition with a high surface/volume ratio and of the chains with an increased external surface/coating available compared with a single nanoparticle or at least one constituent of the composition. In addition to this spatial sequence or arrangement or assembly, we propose to preferentially use a temporal sequence, i.e. to apply an external radiation to the composition or nano-system in an intermittent or sequential manner in order to reduce potential side effects associated with too long and/or too intense continuous radiation application.
The invention relates to the composition or at least one constituent of the composition preferentially comprising at least one nanoparticle or at least one constituent of the composition, wherein preferentially each nanoparticle or at least one constituent of the composition preferentially comprises an iron oxide mineral core or an iron oxide core or a metallic core or a core preferentially composed of a metal oxide, preferentially surrounded by a coating, wherein preferentially the composition or at least one constituent of the composition further comprises a cryo-protectant or protectant compound, wherein preferentially the dissociating energy between the coating and the core is preferentially larger than the dissociating energy between the cryoprotectant or protectant compound and the core, wherein preferentially the core comprises a first center of activity or free radical production or capture, C1FRPC, and/or wherein preferentially the coating comprises a second center of activity or free radical production or capture, C2FRPC, wherein the composition or at least one constituent of the composition is preferentially in the form of a powder or a liquid suspension, wherein C1FRPC and/or C2FRPC preferentially remain(s) comprised or is/are comprised in the core and/or coating, respectively, preferentially at a temperature that is lower than 0° C.
The invention also relates to a nanoparticle or at least one constituent of a composition for use in a method, preferentially method A, for at least one of the following purposes: i) increasing the production of free radicals ii) amplifying radiation, iii) amplifying the effect of radiation, iv) increasing the destruction of the body part, preferentially pathological or tumor cells or a tumor or cancer or virus or a pathological part of the body part, v) destroying or inactivating at least 1, 1.1, 2, 5 or 10 times more pathological or tumor cells or virus preferentially when the pathological cell or pathological body part is exposed to radiation in the presence of the composition than when the pathological cell or pathological body part is exposed to radiation in the absence of the composition, vi) reducing side effects of radiation or preserving the body part, preferentially non-pathological or healthy cells or healthy body part, preferentially surrounding the pathological body part, vii) preserving or maintaining activated or alive at least 1, 1.1, 2, 5 or 10 times more healthy cells preferentially when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition than when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition in the absence of the composition, viii) sonodynamic therapy, ix) photodynamic therapy, x) radiation therapy, xi) a diagnosis, and x) a treatment,
wherein the method preferentially comprises at least one step of:
In one embodiment of the invention, at least one method according to the invention comprises at least 1, 2, 3 or 4 step(s), which is repeated at least 1, 2, 3, 4, 5 or 10 time(s).
In another embodiment of the invention, at least one method according to the invention comprises at least 1, 2, 3 or 4 step(s), which is repeated less than 10, 5, 2 or 1 time(s).
In one embodiment of the invention, during step 2, the quantity of free radicals produced by the radiation is preferentially increased by compounds C and D combined or by compound D alone, wherein during step 3, compound D is preferentially detaching or disassembling from the coating of the nanoparticle or at least one constituent of the composition and is either destroyed/inactivated or diffuses away from the treated body part, wherein during step 4, the quantity of free radicals produced by the radiation is preferentially increased by compound C alone.
In some cases, free radicals can be oxygen or nitrogen species.
The invention also relates to a nanoparticle or at least one constituent of the composition for use in a method, preferentially method B, for at least one of the following purposes: i) decreasing the production of free radicals, ii) reducing radiation, iii) reducing the effect of radiation, iv) increasing the destruction of the body part, preferentially pathological or tumor cells or a tumor or cancer or virus or a pathological part of the body part, v) destroying or inactivating at least 1, 1.1, 2, 5 or 10 times more pathological or tumor cells or virus preferentially when the pathological cell or pathological body part is exposed to radiation in the presence of the composition than when the pathological cell or pathological body part is exposed to radiation in the absence of the composition, vi) reducing side effects of radiation or preserving the body part, preferentially non-pathological or healthy cells or healthy body part, preferentially surrounding the pathological body part, vii) preserving or maintaining activated or alive at least 1, 1.1, 2, 5 or 10 times more healthy cells preferentially when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition than when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition in the absence of the composition, viii) sonodynamic therapy, ix) photodynamic therapy, x) radiation therapy, xi) a diagnosis, and x) a treatment,
wherein the method preferentially comprises at least one step of:
In one embodiment of the invention, during step 2′, the quantity of free radicals or oxygen or nitrogen species produced by the radiation is preferentially decreased by compounds E and F combined or by compound F alone, wherein during step 3′, compound F is preferentially detaching or disassembling from the coating of the nanoparticle or at least one constituent of the composition and is either destroyed/inactivated or diffuses away from the treated body part, wherein during step 4′, the quantity of free radicals or oxygen or nitrogen species produced by the radiation is preferentially decreased by compound E alone.
The invention also relates to nanoparticle or at least one constituent of the composition for sequentially applying radiation on a body part of an individual, comprising a spatial sequence I and/or a temporal sequence II,
wherein the spatial sequence I preferentially consists in applying the radiation on two different centers of activity or free radical production, C1FRP and C2FRP,
wherein C1FRP and C2FRP are preferentially comprised in the core and coating of the nanoparticle or at least one constituent of the composition, respectively,
wherein C1FRP and C2FRP are preferentially spatially separated by more than 0.1 nm, wherein the temporal sequence II preferentially consists in at least one of the following steps:
wherein the method preferentially aims at increasing the quantity of free radicals produced by radiation preferentially during a limited time of exposure of a body part to radiation preferentially to avoid toxicity induced by a too long continuous exposure of a body part to radiation, preferentially when or after nanoparticles or at least one constituent of the composition is(are) introduced in a body part and the radiation is applied on the body part, preferentially sequentially.
The invention also relates to the nanoparticle or at least one constituent of the composition for sequentially applying radiation on a body part of an individual, comprising a spatial sequence III and/or a temporal sequence IV,
wherein the spatial sequence III preferentially consists in applying the radiation on two different centers of activity or free radical capture, C1FRC and C2FRC,
wherein C1FRC and C2FRC are preferentially comprised in the core and coating of the nanoparticle or at least one constituent of the composition, respectively,
wherein C1FRC and C2FRC are preferentially spatially separated by more than 0.1, 1 or 10 nm,
wherein the temporal sequence IV preferentially consists in or comprises at least one of the following steps:
wherein the method preferentially aims at reducing the quantity of free radicals preferentially produced by radiation preferentially by using at least two one or centers free radical capture, preferentially C1FRC and/or C2FRC, and preferentially a limited time of exposure of a body part comprising the nanoparticle or at least one constituent of the composition to radiation.
In some cases, t1, t2, t3, t4, t5, t6, t7 and/or t8 can be larger than 10−10, 10−5, 10−3, 10−2, 10−1, 0, 1, 5, 10 or 103 second(s).
In some other cases, t1, t2, t3, t4, t5, t6, t7 and/or t8 can be smaller than 1010, 105, 103, 102, 10, 0 or 10−3 second(s).
The invention also relates to nanoparticle or at least one constituent of the compositions for use according to the invention, wherein C1FRP and/or C2FRP has/have at least one property selected in the group consisting of:
In some cases, CA1 can be C1FRP or have at least one property in common with C1FRP.
In some other cases, CA2 can be C2FRP or have at least one property in common with C2FRP.
In some cases, C1FRP and C2FRP can be separated by more than 0.1, 1, 5, 10 or 100 nm.
In some other cases, C1FRP and C2FRP can be separated by less than 105, 100, 50, 10, 5, 2, 1 or 0.1 nm.
In some cases, C1FRP and/or C2FRP can be comprised in the nanoparticle or at least one constituent of the composition(s) at a concentration that is larger than 1, 5, 10, 103 or 105 C1FRP and/or C2FRP per nanoparticle or at least one constituent of the composition or per chain of nanoparticle or at least one constituent of the composition.
In some other cases, C1FRP and/or C2FRP can be comprised in the nanoparticle or at least one constituent of the composition(s) at a concentration that is smaller than 105, 103, 10, 5, 2 or 1 C1FRP and/or C2FRP per nanoparticle or at least one constituent of the composition or per chain of nanoparticle or at least one constituent of the composition.
The invention also relates to the nanoparticle or at least one constituent of the composition for use in the method according to the invention, wherein at least two steps among steps 1), 2), 3) and 4) follow each other preferentially in a specific or in any order, i.e. that preferentially: i) step 2 follows step 1), step 2) follows step 3), step 2) follows step 4), step 3) follows step 1), step 3) follows step 2), step 3) follows step 4), step 4) follows step 1), step 4) follows step 2), step 4) follows step 3), step 1) follows step 2), step 1) follows step 3), and/or step 1) follows step 4).
The invention also relates to nanoparticle or at least one constituent of the compositions for use in a method according to the invention, wherein the nanoparticle or at least one constituent of the composition comprises:
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, wherein at least two nanoparticle or at least one constituent of the compositions are organized in a chain.
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, which is characterized by at least one of the following properties:
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, which is characterized by at least one of the following properties:
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein the radiation is selected in the group consisting of: i) an undulating radiation, ii) a particulate radiation, iii) an acoustic wave, iv) a heating radiation, v) a non-heating radiation, vi) an ionizing radiation, vii) a non-ionizing radiation, viii) a laser, ix) a light, x) a sound radiation, xi) an ultrasound radiation, xii) a hyper-sound radiation, xiii) an infrasound radiation, xiv) an X-ray radiation, xv) an electromagnetic radiation, xvi) a radiation due to an electric, magnetic, electromagnetic field, a magnetic field, an alternating magnetic field, xvii) a thermal radiation, xviii) a radiation due to or producing by cold, xix) a radiation due to or producing heat, xx) a radiation produced by or associated with zero-mass, non-zero-mass, neutrons, photon protons, electrons, positron, alpha particle, beta, gamma, neutrinos or muon particle presence or emission, xxi) a radiation oscillating at least 1 frequency or wavelength, and xxii) a source producing any of the previously cited radiation(s).
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein the nanoparticle or at least one constituent of the composition comprises a metallic core that is synthesized by a living organism or nanoparticle or at least one constituent of the composition producing cells, preferentially a magnetosome or magnetosome mineral, wherein the living organism or nanoparticle or at least one constituent of the composition producing cells is preferentially a magnetotactic bacterium.
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein C and/or D is/are photosensitizer(s), selected in the group consisting of: 1) Acridine, such as Acridine Orange, acridine yellow, 2) ALA (5-Aminolevulinic acid), 3) Aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) Aminolevulinic acid, delta-Aminolevulinic acid, 5) Antihistamines, 6) Azulene, 7) Bavteriochlorin, 8) TOOKAD or TOOKAD Soluble, 9) WST-11, 10) LUZ11, 11) BC19, 12) BC21, 13) porphyrin such as Benzoporphyrin derivative monoacid ring A (BPD-MA), 14) Chlorin such as Chlorin e6, m-tetrahydroxyphenylchlorin 15) Foscan, 16) Verteporfin, 17) benzoporphyrin derivative mono acid ring A, 18) Monoaspartyl chlorin(e6), 19) talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrin, 23) Chlorine e6 porphrin, 24) Coal Tar and Derivatives, 25) Contraceptives, Oral and Estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview, 29) Dyes such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squaraines, 33) BODIPY dyes, 34) Phenalenones, 35) benzophenoxazinium dyes, 36) Erythrosine, 37) Flavins, 38) Foscan, 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH), 45) Hypericin, 46) Hypocrellin, 47) ICG (Indocyanine Green), 48) Levulan, 49) MAL-methyl aminolevulinate), 50) Meta-tetra(hydroxyphenyl)chlorin (m-THPC), 51) Metvix, 52) Methylene Blue, 53) Monoterpene, 54) Motexafin lutetium (Lu-Tex), 54) N-aspartyl chlorin e6 (NPe6), 55) Nanoparticle or at least one constituent of the composition or nanomaterial, 56) Natural products or compounds, 57) Non-Steroidal Anti-Inflammatory Drugs, 58) Palladium bacteriopheophorbide (WST09), 59) Phatalocyanin dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phthalocyanine such as Liposomal ZnPC, 65) Chloroaluminium sulfonated phthalocyanine (CASP), 66) Silicon phthalocyanine (PC4), 67) RLP068, 68) Porfimer sodium, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-Tetrakis(1-methylpyridinium-4-yl) porphyrin tosylate, 70) XF70, 71) Protoporphyrin, 72) ALA-induced protoporphyrin IX, 73) Psoralens, 74) Quantum dots, 75) Quinones, 76) Riboflavin, 77) Rose Bengal, 78) silicon or Silicon phthalocyanine (Pc4), 79) Sulfonamides, 80) Sulfonylureas, 81) Talaporfin or Talaporfin soudium, 82) Temoporfin, 82) Tetrahydropyrroles, 83) Tin ethyl etiopurpurin, 84) Titanium dioxide, 85) Toldudine blue O, 86) Transition metal compounds such as Ruthenium(II), polypyridyl complexes, ruthenium, rhodium, cyclometalated, Rh(II)-Rh(II) bridged dimer compounds, platinum(II), gold(III), 87) Verteporfin, 88) Vulnic based compound such as Aminovulinic, aminovulinic acid, 89) WST11, and 90) Xanthene.
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein C and/or D is/are sono-sensitizer(s), selected in the group consisting of: 1) ABS-FA, 2) Acrylonitrile Butadiene Styrene, 3) Styrene, 4) Folic acid, 5) AIMP NP, aminoacyl tRNA synthetase complex-interacting multifunctional protein, 6) Au Nanomaterial, 7) gold, 8) Au—MnO nanomaterial, 9) manganese oxide, 10) Antineoplastic drugs, 11) NSAIDs, 12) nonsteroidal anti-inflammatory drug, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) Au-doped TiO2, 17) Carbon based nanomaterial, 18) carbon nanotube, 19) Chlorine, 20) Ce6, 21) PTX, Paclitaxel, 22) chemotherapeutic drug or compound, 23) infrared dye or IR783, 24) Curcumin, 25) Cyanine or Cu-Cyanine, 26) DHMS, 27) dimethylsulfure, 28) Docetaxel, 29) chemotherapeutic drug or compound, 30) DOX/Mn-TPPS @RBCS, 31) doxorubicin, 32) manganese, 33) blood cell, 34) red blood cell, cell, 35) polymer, 36) elastomer, 37) Erythosin or Erythosin B, 38) FA or FA-OI or FA-OI NP or folic acid, 39) F3-PLGA@MB/Gd NPs, 40) poly(lactic-co-glycolic acid), 41) gadolinium, 42) Fe—TiO2 or titanium oxide, 43) Fe—VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49) hydrochloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysafflor yellow A, 55) Hypocrellin, Hypocrellin B, 56) IR780, 57) Levofloxacin, 58) LIP3 or Lithium phosphide, 59) Lithium, 60) Liposome or Liposomal nanomaterial, 61) Lomefoxacin, 62) MG@P NPs, 63) MnP or Manganese peroxidase, 64) MnTTP-HSAs, 65) HSA-wrapped metal-porphyrin complex, 66) albumin, 67) MnWOx, 68) MnWOx-PEG, 69), PEG, 70) bimetallic oxide, 71) Mn (III)-HFs, 72) managense, hemoporfin, 73) Nanoroads, 74) Noble metal nanomaterial, 75) 01 NP or oxygen indyocyanine, 76) Phthalocyanines, 77) PIO or Pioglitazone, 78) Polymeric nanomaterial, 79) Porphyrin, 80) Pt-doped TiO2, 81) R837, 82) Rose Bengal, 83) Sparfloxacin, 84) TAPP or 5,10,15,20-tetrakis (4-aminophenyl) porphyrin, 85) TiO2 or titanium dioxide nanomaterial, 86) TCPP, isomer, or Tris(1-chloro-2-propyl) phosphate 87) TPI or Thermoplastic Polyimide or thermoplastic polymer, 88) TPZ or Tirapazamine, 89) Transition metal oxide, 90) nanoparticle or at least one constituent of the composition or Janus nanoparticle or at least one constituent of the composition, and 91) Xanthones.
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein C and/or D is/are radio-sensitizer(s), selected in the group consisting of: 1) AMG102, 2) AQ4N, 3) Apaziquone (E09), 4) Bromodeoxyuridine, 5) Carbogen, 6) Cetuximab, 7) Chemotherapeutic drug or compound, 8) Chlorpromazine, 9) C-reactive peptide, 10) Curcumin, 11) Diamide, 12) Diethylmaeate, 13) Dihydroartemisinin, 14) Docetaxel, 15) ECI301, 16) Etanidazole, 17) Fludarabine, 18) 5-Fluorouracil, 19) Fluorodeoxyuridine, 20) Gadolynium, 21) Gemcitabine, 22) HER-3 ADC, 23) HSP, 24) Hydrogen peroxide, 25) Hydroxyurea, 26) Hyperbaric oxygen, 27) Hyperthermia, 28) Hypoxic cell cytotoxic agent, 29) Irinotecan, 30) lanthanide-doped radiosensitizer-based metal-phenolic network, 31) Lidocaine, 32) Lododeoxyuridine, 33) Metronidazole, 34) misonidazole, 35) etanidazole, 36) nimorazole, 37) N-Ethylmalemide, 38) malmeide, 39) ethylmalmeide, 40) Nanomaterial such as those consisting of or composed of at least partly or fully gold, silver, bismuth, gadolinium, polysiloxane matrix and gadolinium chelates, hafnium, Tantalum, Zinc, Gadolinium, Germanium, Chromium, Praseodymium, Silicon, iron, platinum, cobalt, manganese, magnesium, iron, Titanium, carbon nanotube, quantum dot, nanoroad, Triflate, or metal oxide, 41) Nelfinavir, 42) Nicotinamide, 43) Nimotuzumab, 44) RNA, or miRNA, or miR-201, or miR-205, or miR-144-5p, or miR-146a-5p, or miR-150, or miR-99a, or miR-139-5p, or miR-320a, 45) Membrane active agent, 46) Mitomycin-C or Mitomycin, 47) Motexafin, 48) NBTXR3, 49) Oligonucleotide, 50) Paclitaxel, 51) Papaverine or Papaverine hydrochloride, 52) Paraxonase-2, 53) Pocaine, 54) Porfiromycin (POR), 55) Protein, 56) Peptide, 57) Radiosensitizing nucleosides or compounds, 58) Resveratrol, 59) RRx-001, 60) SiRNa, 61) Suppressors of sulfhydral groups, 62) SYM004, 63) Texaphyrins, 64) TH-302, and 65) Tirapazamine.
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein C and/or D is/are oxidizing compound(s).
The invention also relates to nanoparticle or at least one constituent of the composition for use in a method according to the invention, wherein C and/or D is/are anti-oxidizing compound(s).
The inventions relates to a preparation comprising at least one multiply active nanoparticle or at least one constituent of the composition, where the nanoparticle or at least one constituent of the composition comprises:
In some cases, the center of activity or free radical capture CFRC1 or CFRC2 comprises a compound E.
The invention relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, wherein the coating of the nanoparticle or at least one constituent of the composition preferentially comprises holes, enabling the diffusion of free radicals produced by compound C and/or A preferentially during step 2) or the capture of free radicals by the compound E preferentially during step 2′), hence preferentially increasing the quantity of produced free radicals by compound(s) C and/or A or the quantity of free radicals captured by compound E.
The invention also relates to nanoparticles or at least one constituent of the composition for use in the method according to the invention to detach or disassemble the coating of the nanoparticle or at least one constituent of the composition, compound C or compound E, preferentially by increasing the duration t2 or t5, or by choosing a duration t2 or is that is longer than t1, t4, t3, and/or t6, or by introducing the nanoparticle or at least one constituent of the composition(s) in a degrading/altering medium/material such as acidic/basic pH media or liposomes.
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, wherein the nanoparticle or at least one constituent of the composition comprises:
The invention also relates to nanoparticle or at least one constituent of the compositions for use according to the invention, where the central part or core of the nanoparticle or at least one constituent of the composition has at least one property selected in the group consisting of:
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, where the compound C is selected among zinc and aluminum.
The invention also relates to the nanoparticle or at least one constituent of the compositions for use in a method according to the invention, wherein the proportion of A, B, and C compounds in the central part or core of the nanoparticle or at least one constituent of the composition is such that:
The invention also relates to the nanoparticle or at least one constituent of the composition for use in the method according to the invention, wherein the coating part surrounding the central part or core of the nanoparticle or at least one constituent of the composition has at least one property selected in the group consisting of:
Nanoparticle or at least one constituent of the composition for use in a method according to any of the claims 1 to 9 The preparation according to claim 8, where C and D compounds act synergistically, i.e. that the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising C and D is larger than the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising only C or only D.
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, where C and D are compounds selected in two different groups 1 and 2, where:
The invention also relates to nanoparticle or at least one constituent of the composition for use in the method according to the invention, where C and D act anti-synergistically, i.e. that the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising C and D is lower than the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising only C or only D.
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention, where the nanoparticle or at least one constituent of the composition first comprises the compounds C and D during a time t7 and second comprises the compounds D during a time t8, where is preferentially follows t7.
The invention also relates to nanoparticle or at least one constituent of the compositions for use in the method according to the invention being or comprising: i) a medical device, ii) a drug, iii) a pharmaceutical product or composition or at least one constituent of the composition, iv) a medical product or composition or at least one constituent of the composition, iv) a biological product or composition or at least one constituent of the composition, v) a composition or at least one constituent of the composition, vi) an adjuvant, vii) an excipient, viii) an active principle, ix) a vaccine or vaccine component, and/or vi) a product.
The invention also relates to the nanoparticle or at least one constituent of the compositions for use in a method according to the invention.
The invention relates to the method according to the invention for use in photodynamic therapy, sonodynamic therapy, and/or radiotherapy.
The invention also relates to nanoparticle or at least one constituent of the composition for use in the method according to the invention, for the treatment of an infectious, viral, or cancerous disease.
The invention also relates to a method of fabrication of the nanoparticle or at least one constituent of the composition according to the invention, which comprises at least one of the following steps:
The invention also relates to the method according to the invention, wherein the concentration of the source of zinc, preferentially zinc citrate or zinc sulfate, is comprised between 1 and 100 μM, preferentially 2 and 50 μM, most preferentially 5 and 20 μM.
The invention also relates to the method according to the invention, wherein the nanoparticle or at least one constituent of the compositions fabricated according to the method according to the invention comprise a quantity of metal other than iron, preferentially zinc or aluminum, which is incorporated in or comprised in the metallic core is: i) comprised between 0.1 and 100 mg of metal other than iron comprised in the core per gram of iron comprised in the core, ii) preferentially comprised between 0.5 and 10 mg of metal other than iron comprised in the core per gram of iron comprised in the core, iii) most preferentially comprised between 1 and 5 mg of metal other than iron comprised in the core per gram of iron comprised in the core.
The invention relates to a composition or at least one constituent of the composition comprising at least one nanoparticle or at least one constituent of the composition, wherein each nanoparticle or at least one constituent of the composition preferentially comprises an iron oxide mineral core or an iron oxide core or a metallic core or a core composed of a metal oxide, surrounded by a coating,
In some cases, the nanoparticle or at least one constituent of the composition comprises a core and/or a coating, wherein the coating preferentially stabilizes the core and/or prevents the core to aggregate and/or leads to the chain arrangement of at least two nanoparticle or at least one constituent of the compositions.
In some other case, the nanoparticle or at least one constituent of the composition is amorphous and/or crystallized.
In some cases, an iron oxide core is composed of at least one iron atom and at least one atom of oxygen.
In some other cases, a core composed of a metal oxide is composed of at least one metal atom and at least one oxygen atom.
In still some other cases, a metallic core is composed of at least one metal atom.
In one embodiment of the invention, the composition or at least one constituent of the composition comprises the at least one nanoparticle or at least one constituent of the composition with the cryoprotectant or protectant compound and optionally a liquid such as water.
In some cases, the nanoparticle or at least one constituent of the composition comprises a core and/or a coating, wherein the coating preferentially stabilizes the core and/or prevents the core to aggregate and/or leads to the chain arrangement of at least two nanoparticle or at least one constituent of the compositions.
In some other case, the nanoparticle or at least one constituent of the composition is amorphous and/or crystallized.
In some cases, an iron oxide core is composed of at least one iron atom and at least one atom of oxygen.
In some other cases, a core composed of a metal oxide is composed of at least one metal atom and at least one oxygen atom.
In still some other cases, a metallic core is composed of at least one metal atom.
In one embodiment, the center of activity or free radical production or capture is a compound that increases the quantity of free radicals produced or captured when: i) a radiation is applied on the center, preferentially designated as radio-sensitizer in this case, ii) light is applied on the center, preferentially designated as photo-sensitizer in this case, iii) thermal radiation or temperature increase or temperature decrease is applied on the center, preferentially designated as thermo-sensitizer in this case, iv) cryo treatment or protection applied on the center, preferentially designated as cryo-sensitizer in this case, v) a magnetic field is applied on the center, preferentially designated as magneto-sensitizer in this case, vi) ultrasound is applied on the center, preferentially designated as sonosensitizer.
In one embodiment, the center of activity or free radical production or capture can be a center of physico-chemical disturbance, such as radiation, amplification, where the amplification can correspond to an effect of the physico-chemical disturbance applied on the body part that is more important in the presence than in the absence of the center, for example eradication or death of a tumor could be induced in the presence of the irradiated center while such effect would be absent or less pronounced in the absence of the center.
In one embodiment, the center of activity or free radical production or capture is a compound that amplifies or increases the physico-chemical disturbance or radiation or at least one parameter or property of the physico-chemical disturbance or radiation or at least one effect of a radiation or physico-chemical disturbance. When the center is exposed to radiation, light, thermal variation, cryo-treatment or cryo-protection, magnetic field, or ultrasound, it can be designated as radio-sensitizer, photo-sensitizer, thermo-sensitizer, cryo-sensitizer, magneto-sensitizer, or sonosensitizer, respectively.
In another embodiment of the invention, the composition or at least one constituent of the composition comprises the at least one nanoparticle or at least one constituent of the composition without the cryoprotectant or protectant compound and optionally a liquid such as water.
In another embodiment of the invention, the composition or at least one constituent of the composition comprises an excipient, an active principle, and/or a surfactant.
In another embodiment of the invention, the composition or at least one constituent of the composition is a preparation or a suspension or a powder, preferentially of or with the at least one nanoparticle or at least one constituent of the composition, optionally with the cryo-protectant or protectant compound.
In one embodiment of the invention, the cryo-protectant or protectant compound is or comprises at least one substance or compound or chemical function or molecule or atom, which protects or maintains or keeps at least one property of the composition or at least one constituent of the composition, preferentially selected among: i) the chain arrangement or geometric figure of the at least two nanoparticles or at least one constituent of the composition(s), ii) the at least one center of activity or free radical capture or production, iii) the isotonicity or osmolality, preferentially of or in the composition or at least one constituent of the composition, preferentially at or below the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, −5, −10, −50, −77, −100, −200 or −273° C.
In another embodiment of the invention, the at least one property of the composition or at least one constituent of the composition is maintained, or kept or protected when:
preferentially for or when the temperature of the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition is maintained at or decreased below or at the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, −5, −10, −50, −77, −100, −200 or −273° C.
In some cases, T1 and T2 are two different temperatures.
In some cases, T2 is smaller than T1, preferentially by at least 10−5, 10−3, 10−1, 0, 1, 2, 5, 10 or 103° C.
In some cases, T1 and/or T2 is/are smaller than 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, −5, −10, −50, −77, −100, −200 or −273° C.
In some other cases, T1 and/or T2 is/are larger than −273, −200, −100, −77, −50, −10, −5, 0, 1, 2, 5, 10, 50, 100 or 103° C.
In one embodiment of the invention, the composition or at least one constituent of the composition can be cooled down, preferentially below or at 10, 5, 2, 1, 0, −5, −10, −50, −77, −90, −270, or −273° C., or the composition or at least one constituent of the composition below or at 10, 5, 2, 1, 0, −5, −10, −50, −77, −90, −270, or −273° C., can be reached or obtained preferentially for more than 1 second, 1 minute or 1 hour, preferentially without destroying at least one chain or geometric figure or at least one center of activity or free radical/production, preferentially with at least one chain or geometric figure or at least one center of activity or free radical/production that is maintained or exists. This effect is preferentially due to the presence of the cryo-protectant or protectant compound.
In one embodiment of the invention, the composition or at least one constituent of the composition is lyophilized or desiccated or dehydrated, preferentially for more than 1 second, 1 minute or 1 hour, preferentially without destroying or removing at least one center of activity or free radical capture or production or at least one chain or geometric figure, preferentially with at least one chain or geometric figure at least one center of activity or free radical/production that is maintained or exists. This effect is preferentially due to the presence of the cryo-protectant or protectant compound.
In one embodiment of the invention, the composition or at least one constituent of the composition is prepared or used by following at least one of the following steps:
In one embodiment, the at least one step mentioned above or of a method according to the invention is repeated.
In one embodiment of the invention, the dissociation energy between a chemical group of a molecule of the coating and a chemical group of the core of the nanoparticle or at least one constituent of the composition is larger than 10−5, 1, 0, 10 or 100 Kcal, KJ, or eV, preferentially per mol or per bond.
In one embodiment of the invention, the dissociation energy between a chemical group of a molecule of the coating and a chemical group of the core of the nanoparticle or at least one constituent of the composition is preferentially larger than 10 Kcal/mol, 100 KJ/mol, or 1 eV/bond.
In one embodiment of the invention, the dissociation energy between: i) a chemical group of a molecule of the cryo-protectant or protectant compound and a chemical group of the core of the nanoparticle or at least one constituent of the composition or ii) a chemical group of a molecule of the cryo-protectant or protectant compound and a chemical group of the coating of the nanoparticle or at least one constituent of the composition, is lower than 1020, 105, 103, 100, 10, 1 or 0 Kcal, KJ, or eV, preferentially per mol or per bond.
In one embodiment of the invention, the dissociation energy between: i) a chemical group of a molecule of the cryoprotectant or protectant compound and a chemical group of the core of the nanoparticle or at least one constituent of the composition or ii) a chemical group of a molecule of the cryoprotectant or protectant compound and a chemical group of the coating of the nanoparticle or at least one constituent of the composition, is preferentially lower than 10 Kcal/mol, 100 KJ/mol, or 1 eV/bond.
The invention relates to the composition or at least one constituent of the composition according to invention, wherein the chemical affinity between the coating and the core of the nanoparticle or at least one constituent of the composition is preferentially is larger than: i) the chemical affinity between the cryoprotectant or protectant compound and the coating and/or ii) the chemical affinity between the cryoprotectant or protectant compound and the core of the nanoparticle or at least one constituent of the composition. In one embodiment of the invention, the coating is more strongly bound to or is in stronger interaction with the nanoparticle core or at least one constituent of the composition core than the cryo-protectant or protectant compound.
In one embodiment of the invention, the coating can't be separated or isolated from the nanoparticle core or at least one constituent of the composition core or is inseparable or un-isolable from the nanoparticle core or at least one constituent of the composition, preferentially using a magnet or magnetic separation or centrifugation.
In another embodiment of the invention, the cryo-protectant or protectant compound can be separated or isolated from the nanoparticle core or at least one constituent of the composition core or is separable or isolable from the nanoparticle core or at least one constituent of the composition core, preferentially using a magnet or magnetic separation or centrifugation.
In some cases, this property can be highlighted by the fact that: i) the coating and core of the nanoparticle or at least one constituent of the composition can't be separated by a magnet of low intensity, i.e. preferentially of intensity smaller than 1 T or 1 mT, or of low magnetic field gradient, i.e. preferentially of magnetic field gradient smaller than 1 T or mT per cm preferentially of body part, whereas the cryoprotectant or protectant compound can preferentially be separated from the nanoparticle or at least one constituent of the composition by such magnet, ii) the coating and core of the nanoparticle or at least one constituent of the composition can't preferentially be separated by mixing the nanoparticle core or at least one constituent of the composition and coating in water followed by centrifugation whereas the cryoprotectant or protectant compound can preferentially be separated from the nanoparticle or at least one constituent of the composition by being mixed in water with the suspended nanoparticle or at least one constituent of the compositions followed by centrifugation.
In one embodiment, the coating or at least one constituent of the composition comprises at least one compound or chemical function, which is able to establish interactions or weak interactions or weak bonds or covalent bonds with the core of the nanoparticle or at least one constituent of the composition or at least one chemical function or atom or ion of the core part of the nanoparticle or at least one constituent of the composition, in particular iron oxide or any combination or ionic state of iron and/or oxygen. In some cases, such interactions or bonds maintain the nanoparticle coating or at least one constituent of the composition attached or linked or associated with or to the nanoparticle core or at least one other constituent of the composition, preferentially called associating or attaching or linking bonds or interactions.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition consists of: a core, preferentially a surrounding coating and/or preferentially associating or linking or attaching bonds or interactions.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition consists of a core and/or a surrounding coating, which is preferentially associated or linked or attached to or with the core, preferentially in such a way or through sufficiently strong bonds or interactions that it does not detach or dissociates from the core or that it remains attached or bound to the core.
In some cases, the at least one center of activity or free radical capture or production or chain arrangement or geometric figure of the nanoparticle or at least one constituent of the compositions is maintained or exists or forms in the presence of the coating.
In some cases, in the absence of coating the at least one center of activity or free radical capture or production or chain arrangement or geometric figures does not exist or does not form or does not exist.
In one embodiment of the invention, the composition or at least one constituent of the composition comprises a cryo-protectant or protectant compound, which is associated or linked or bound with or to the nanoparticle or at least one other constituent of the composition, core, coating, preferentially in such a way or through sufficiently weak bonds or interactions that it detaches or dissociates from the nanoparticle or at least one other constituent of the composition, core or coating, preferentially when the cryo-protectant or protectant compound is washed away or removed from the nanoparticle or at least one other constituent of the compositions with water, by centrifugation or by using a magnet.
In some cases, the cryo-protectant or protectant compound can be dissociated or detached or disassembled or unlinked from the nanoparticle or at least one other constituent of the composition, core or coating, preferentially in such a way that the at least one center of activity or free radical capture or production or chain arrangement or geometry of the nanoparticle or at least one constituent of the compositions is maintained optionally comprised in the nanoparticle or at least one other constituent of the composition.
In some cases, the cryo-protectant or protectant compound can be associated or attached or assembled or linked to or with the nanoparticle or at least one other constituent of the composition, core or coating, preferentially in such a way that the at least one center of activity or free radical capture or production or chain arrangement or geometry of the nanoparticle or at least one constituent of the compositions is maintained, optionally in the nanoparticle or at least one constituent of the composition, preferentially when the composition or at least one constituent of the composition is cooled down, preferentially below or at 100, 10, 5, 2, 1, 0, −10, −50, −77, −270, −273° C., preferentially for more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the composition or at least one constituent of the composition is lyophilized or desiccated or when water is removed from the composition or at least one constituent of the composition partly or fully or when the percentage in mass of water in the composition or at least one constituent of the composition is lower than 100, 50, 10, 5, 2, 1, 0, 10−3 or 10-5% or when the composition or at least one constituent of the composition is in powder form.
In some cases, the at least one center of activity or free radical capture or production or chain arrangement or geometric figure of the nanoparticle or at least one constituent of the compositions can be observed by following at least one of the following step: i) removing the cryoprotectant or protectant compound from the composition or at least one constituent of the composition, ii) suspending or resuspending the composition or at least one constituent of the composition, preferentially lyophilized, preferentially in water, iii) by depositing a droplet of the composition or at least one constituent of the composition on top of a substrate, preferentially a carbon grid, iv) by waiting for water evaporation, and v) by observing the nanoparticle or at least one constituent of the composition arrangement under electron microscopy.
In some cases, preferentially in the absence of coating at least one center of activity or free radical capture or production the chains or geometric figures do not form or exist while in the presence of coating the chains or geometric figures or at least one center of activity or free radical/production preferentially form or exist.
In some cases, when the temperature of the at least one nanoparticle or at least one constituent of the composition is lower than or equal to 100, 10, 5, 2, 1, 0, −10, −50, −77, −270, −273° C., preferentially for more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the nanoparticle or at least one constituent of the composition is lyophilized or desiccated or dehydrated or dried or when water is removed from the composition or at least one constituent of the composition or from the at least one nanoparticle or at least one constituent of the composition partly or fully or when the percentage in mass of water in the composition or at least one constituent of the composition the at least one nanoparticle or at least one constituent of the composition is lower than 100, 50, 10, 5, 2, 1, 0, 10−3 or 10−5%, at least one center of activity or free radical capture or production or the chain arrangement or the geometric figure or assembly preferentially does not exist or does not form or is destroyed in the absence of the cryo-protectant or protectant compound, and/or the at least one center of activity or free radical capture or production or chain arrangement or the geometric figure or assembly preferentially exists or forms or is maintained or is not destroyed in the presence of the cryo-protectant or protectant compound.
In one embodiment of the invention, the coating or at least one compound or chemical function comprised in the coating is or remains chemisorbed or physisorbed or adsorbed or attached to or linked to or associated with or bound to the core of the nanoparticle or at least one constituent of the composition, preferentially in the presence of the cryoprotectant or protectant compound, preferentially surrounding or embedding the nanoparticle or at least one constituent of the composition, preferentially for a duration of more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, preferentially for or when the composition or at least one constituent of the composition is in powder form, preferentially for or when the percentage in mass of water in the composition or at least one constituent of the composition is lower than 100, 50, 10, 5, 2, 1, 0, 10−3 or 10−5%.
In one embodiment of the invention, the coating or at least one compound or chemical function comprised in the coating is not or does not remain chemisorbed or physisorbed or adsorbed or attached to or linked to or associated with or bound to the core of the nanoparticle or at least one constituent of the composition, preferentially in the absence of the cryoprotectant or protectant compound, preferentially surrounding or embedding the nanoparticle or at least one constituent of the composition, preferentially for a duration of more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, preferentially for or when the composition or at least one constituent of the composition is in powder form, preferentially for or when the percentage in mass of water in the composition or at least one constituent of the composition is lower than 100, 50, 10, 5, 2, 1, 0, 10−3 or 10−5%.
In one embodiment, the coating or at least one compound or chemical function comprised in the coating is linked through bonds or interactions with or to Fe2+ or Fe3+ ions, hydroxyls OH−, oxides O2−, crystalline defects of the core, which may be in or at the surface of the core of the nanoparticle or at least one constituent of the composition.
In one embodiment, the coating comprises at least one compound, atom, ion, or chemical function such as an acid, carboxylic acid, phosphoric acid, or sulfonic acid function, wherein the compound, atom or ion contained in the coating is able to establish interactions or bonds with the core or with at least one atom of the core, a chemical function of the core, an ion of the core such as Fe2+, Fe3+, Hydroxyl OH−, oxide O2− or a crystalline defect of the core.
In one embodiment of the invention, the interactions or bonds exist or are maintained when the composition or at least one constituent of the composition is cooled down, preferentially below or at 100, 10, 5, 2, 1, 0, −10, −50, −77, −270, −273° C., preferentially for more than 0.001, 0.1, 0, 1, 5, 10, 103 or 105 seconds, or when the composition or at least one constituent of the composition is lyophilized or desiccated or dehydrated or dried or when water is removed from the composition or at least one constituent of the composition partly or fully or when the percentage in mass of water in the composition or at least one constituent of the composition is lower than 100, 50, 10, 5, 2, 1, 0, 10−3 or 10−5% or when the composition or at least one constituent of the composition is in powder form, preferentially in the presence of the cryoprotectant or protectant compound. In some cases, these interactions or bonds keep the at least one center of activity or free radical capture or production comprised in the nanoparticle or at least one constituent of the composition or nanoparticle or at least one constituent of the compositions organized in chain or in geometric figure, i.e. that preferentially in the absence of these interactions or bonds, the nanoparticle or at least one constituent of the compositions are not organized in chain or geometric figure or the at least one center of activity or free radical capture or production is not comprised in the nanoparticle or at least one constituent of the composition.
In some cases, a geometric figure is an assembly or aggregate of nanoparticle or at least one constituent of the compositions forming a geometric figure.
In some cases, a geometric figure is selected from the group consisting of: a Balbis, Concave polygon, Constructible polygon, Convex polygon, Cyclic polygon, Equiangular polygon, Equilateral polygon, Penrose tile, Polyform, Regular polygon, Simple polygon, Tangential polygon, Polygons with specific numbers of sides, Henagon, Digon, Triangle, Acute triangle, Equilateral triangle, Heptagonal triangle, Isosceles triangle, Obtuse triangle, Rational triangle, Right triangle, Kepler triangle, Scalene triangle, Quadrilateral, Cyclic quadrilateral, Kite, Parallelogram, Rhombus, Lozenge, Rhomboid, Rectangle, Square, Tangential quadrilateral, Trapezoid, Isosceles trapezoid, Pentagon, Hexagon, Lemoine hexagon, Heptagon, Octagon, Nonagon, Decagon, Hendecagon, Dodecagon, Tridecagon, Tetradecagon, Pentadecagon, Hexadecagon, Heptadecagon, Octadecagon, Enneadecagon, Icosagon, Swastika, Star polygon, Pentagram—star polygon, Hexagram, Star of David, Heptagram, Octagram, Star of Lakshmi, Decagram—star polygon, Annulus, Arbelos, Circle, Archimedes' twin circles, Bankoff circle, Circumcircle, Disc, Incircle and excircles of a triangle, Nine-point circle, Circular sector, Circular segment, Crescent, Indalo, Lens, Lune, Reuleaux polygon, Reuleaux triangle, Salinon, Semicircle, Tomahawk, Triquetra, Heart, Archimedean spiral, Astroid, Cardioid, Deltoid, Ellipse, Heart, Heartagon, Various lemniscates, Oval, Cartesian oval, Cassini oval, Oval of Booth, Ovoid, Superellipse, Taijitu, Tomoe, and/or Magatama.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition, the suspension, composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the composition is stable, preferentially during a lapse of time, preferentially being its stability duration, which is larger than 10−10, 5, 10, 1050 or 10100 minute(s), 1, 2, 3, 4, 5, 6, 10, 12, 24 or 36 months.
In some cases, the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions can be stable preferentially at a concentration of nanoparticle or at least one constituent of the compositions larger than 1, 5, 10, 50, 100, 200, 500 or 1000 mg of nanoparticle or at least one constituent of the compositions per mL of solvent, water, matrix, or body part surrounding or comprising or embedding the or nanoparticle or at least one constituent of the compositions or composition or at least one constituent of the composition.
In one embodiment of the invention, the body part is the body part of an individual, animal, or human.
In an embodiment of the invention, the body part comprises more than or at least 1, 2, 5, 10, or 100 similar or different organism(s), apparatus, organ(s), tissue(s), cell(s), or biomolecule(s).
In some cases, the body part can be all or part of the head, neck, shoulder, arm, leg, knee, foot, hand, ankle, elbow, trunk, inferior members, or superior members.
In some other cases, the body part can be or belong to an organ, the musculoskeletal, muscular, digestive, respiratory, urinary, female reproductive, male reproductive, circulatory, cardiovascular, endocrine, circulatory, lymphatic, nervous (peripheral or not), ventricular, enteric nervous, sensory, or integumentary system, reproductive organ (internal or external), sensory organ, endocrine glands. The organ or body part can be human skeleton, joints, ligaments, tendons, mouth, teeth, tongue, salivary glands, parotid glands, submandibular glands, sublingual glands, pharynx, esophagus, stomach, small intestine, duodenum, jejunum, ileum, large intestine, liver, gallbladder, mesentery, pancreas, nasal cavity, pharynx, larynx, trachea, bronchi, lungs, diaphragm, kidneys, ureters, bladder, urethra, ovaries, fallopian tubes, uterus, vagina, vulva, clitoris, placenta, testes, epididymis, vas deferens, seminal vesicles, prostate, bulbourethral glands, penis, scrotum, pituitary gland, pineal gland, thyroid gland, parathyroid glands, adrenal glands, pancreas, heart, arteries, veins, capillaries, lymphatic vessel, lymph node, bone marrow, thymus, spleen, gut-associated lymphoid tissue, tonsils, brain, cerebrum, cerebral hemispheres, diencephalon, brainstem, midbrain, pons, medulla, oblongata, cerebellum, spinal cord, choroid plexus, nerves, cranial nerves, spinal nerves, ganglia, eye, cornea, iris, ciliary body, lens, retina, ear, outer ear, earlobe, eardrum, middle ear, ossicles, inner ear, cochlea, vestibule of the ear, semicircular canals, olfactory epithelium, tongue, taste buds, mammary glands, or skin.
In some cases, the body part or organ can belong to the blood circulation or circulatory system.
In some cases, the body part can be or comprise at least one tumor, cancer, virus, bacterium, or pathological cell.
In one embodiment of the invention, the body part is or comprises water, an excipient, a solution, a suspension, at least one chemical element, organic material, or gel, which can be synthetic or produced by a living organis.
Preferably, the body part of an individual, also designated as the body part, represents or is part of an individual or a whole individual, where the individual is preferentially a human, an animal, or an organism, preferentially a living or inactivated or dead organism, comprising at least one prokaryotic or eukaryotic cell.
In one embodiment of the invention, the body part is alive (or not), is any tissue, water, medium, substance, cell, organelle, organ protein, lipid, DNA, RNA, biological material, preferentially localized in a specific region of an individual, preferentially originating or extracted from such region.
In an embodiment of the invention, the body part comprises a pathological site, a healthy site, and/or a nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition region.
In one embodiment of the invention, the body part is or comprises a pathological site or pathological cells.
In some cases, the pathological site can be defined as an unhealthy site, or a site that is in a different condition from a site of a healthy individual, or the site of an unhealthy individual. It can comprise pathological cells, such as tumor cells, bacteria, eukaryotic or prokaryotic cells, as well as viruses or other pathological material. Pathological cells can be cells that are: i) not arranged or working as they usual do in a healthy individual, ii) dividing more quickly than healthy cells, iii) healthy cells having undergone a transformation or modification, iv) dead, sometimes due to the presence of a virus or to other organisms, or v), in contact, in interaction, with foreign material not belonging to the individual, such as viruses, where viruses can possibly penetrate, colonize, or replicate in these cells. In some cases, pathological cells can be assimilated to viruses or to other organisms or entities that colonize cells or target cells or destroy cells or use cells or enter in interaction with cells, preferentially to enable their own reproduction, multiplication, survival, or death. In some cases, a pathological site can comprise healthy cells, preferentially with a lower number, activity or proliferation, than that of pathological cells.
In one embodiment of the invention, the body part is or comprises a healthy site or healthy cells. In some cases, the healthy site can be defined as a site or region that comprises healthy cell(s), where a healthy cell can be defined as a cell that belongs to a healthy individual or to the body part of a healthy individual.
In some cases, the healthy site can surround the pathological site when it is preferentially located at a distance of less than 1 or 10−9 m from the pathological site.
In some cases, the composition or at least one constituent of the composition or body part can be exposed to radiation.
Preferably, the radiation is selected from the group consisting of: i) a magnetic or electric or electromagnetic field or wave, a wave a particulate radiation, ii) laser light, iii) light produced by a lamp, iv) light emitted at a single wavelength, v) light emitted at multiple wavelengths, vi) a ionizing radiation, vii) microwave, viii) radiofrequencies, and ix) a sound, an ultrasound an infrasound, or an acoustic wave. In one embodiment of the invention, the radiation has at least one of the following properties:
In another embodiment of the invention, the radiation has at least one of the following properties:
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition is administered to or in the body part.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition is administered at a distance of less than 1 or 10−9 m away from the body part.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition is administered at a distance of more than 1 or 10−9 m away from the body part.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition is administered to or in the body part preferentially following at least one of the following administration routes: local, enteral, gastrointestinal, parenteral, topical, oral, inhalation, intramuscular, subcutaneous, intra-tumor, in an organ, in a vein, in arteries, in blood, or in tissue.
In some cases, the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, the composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions can be stable preferentially when at least one nanoparticle or at least one constituent of the composition is not degraded or does not lose partly or fully its coating or can be administered to a body part or keeps or maintains its chain arrangement or geometric figure or the presence of at least one center of activity or free radical/production.
In some other cases, the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, the composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions can be stable when the optical density of the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the composition, preferentially mixed in water, preferentially measured at 480 nm or at another fixed wavelength, does not decrease preferentially by more than 1, 5, 10, 50, 75 or 90% or by more than 10−10, 10−3, 10−1, 0.5 or 0.7, preferentially within 1, 5, 10, 103, 107 or 1020 seconds following homogenization or mixing or optical density measurement or absorption measurement of this suspension or composition or at least one constituent of the composition. This percentage can be equal to (ODB−ODA)/ODB or ODA/ODs, where ODs is the optical density of the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, the composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions measured before the homogenization or mixing or optical density measurement or absorption measurement of the nanoparticle or at least one constituent of the composition, the suspension, composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the composition and ODA is the optical density of the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions measured after the homogenization or mixing or optical density measurement or absorption measurement of the nanoparticle or at least one constituent of the compositions, the suspension of nanoparticle or at least one constituent of the compositions, the composition or at least one constituent of the composition, or assembly of nanoparticle or at least one constituent of the compositions.
In some cases, the composition or at least one constituent of the composition can be stable or considered stable preferentially when it is measured stable at a certain first time to and at a certain second time t1, where t1 follows to or is separated from to by a lapse of time ΔT of at least 1 second, 1 minute, 1 hour, 1 day, 1 month, 3 months, 6 months, 1, 2, 5, or 10 years. During ΔT, the composition or at least one constituent of the composition is preferentially being stocked.
In some cases, the nanoparticle or at least one constituent of the composition can be suspended in a liquid or dispersed in a matrix or body part preferentially to yield a homogenous nanoparticle or at least one constituent of the composition dispersion or a highly stable nanoparticle or at least one constituent of the composition or suspension.
In one embodiment of the invention, the cryoprotectant or protectant compound serves to maintain the chain or geometric figure or at least one center of activity or free radical/production stable over a period of time, preferentially ΔT, preferentially at least one or six months. In this case, the chain or geometric figure or at least one center of activity or free radical/production can preferentially be observed or measured, preferentially under electron microscopy measurements, at a first time to and at a second time t1, where t1 is preferentially separated from to by ΔT.
In one embodiment of the invention, the cryoprotectant or protectant compound protects or maintains the arrangement of at least two nanoparticle or at least one constituent of the compositions in at least one chain or in at least one geometric figure or the presence of at least one center of activity or free radical/production in the composition or at least one constituent of the composition.
In some cases, the protection or maintenance of the arrangement of at least two nanoparticle or at least one constituent of the compositions in at least one chain or in at least one geometric figure or the presence of at least one center of activity or free radical/production occurs at a temperature preferentially of the composition or at least one constituent of the composition that is preferentially lower than or equal to 103, 500, 100, 50, 10, 5, 2, 1, 0, −5, −10, −20, −50 −100, −200, or −273° C.
In some other cases, the protection or maintenance of the arrangement of at least two nanoparticle or at least one constituent of the compositions in at least one chain or in at least one geometric figure or the presence of at least one center of activity or free radical/production occurs at a temperature preferentially of the composition or at least one constituent of the composition that is preferentially larger than or equal to −273, −200, −100, −50 −20, −10, −5, 0, 1, 2, 5, 10, 50, 100, 500, or 103° C.
In some cases, the protection or maintenance of the arrangement of at least two nanoparticle or at least one constituent of the compositions in at least one chain or in at least one geometric figure or of the presence of at least one center of activity or free radical/production occurs for a duration that is preferentially larger than 10−10, 10−5, 10−3, 10−1, 0, 1, 2, 5, 10, 102, 103 or 105 minutes or seconds.
In some other cases, the protection or maintenance of the arrangement of at least two nanoparticle or at least one constituent of the compositions in at least one chain or in at least one geometric figure or of the presence of at least one center of activity or free radical/production occurs for a duration that is preferentially smaller than 1010, 105, 103, 10, 5, 2, 1, 0, 10−1, 10−3 or 10−5 minutes or seconds.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are preferentially selected in the group consisting of:
In some cases, C1FRPC and/or C2FRPC can comprises at least one atom that is preferentially linked to at least one oxygen or iron atom of the core preferentially through at least one metallic bound.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein coating comprises a second center of activity or free radical production or capture C2FRPC.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C2FRPC is linked to at least one atom of the coating, preferentially through at least one covalent bond.
In one embodiment of the invention, the center a free radical production or capture is at least one atom, molecule, chemical function, ionized or not, charged or not, which produces or captures at least one free radical.
In one embodiment of the invention, a free radical is at least one atom, molecule, ion or chemical function that has: i) at least one unpaired valence electron, ii) a capacity to dimerize, iii) a short lifetime, and/or iv) two unpaired electrons.
In one embodiment of the invention, a free radical is at least one atom, molecule, ion or chemical function that is: i) a chemically reactive species, ii) a hydroxyl radical HO., iii) a triplet oxygen, iv) a triplet carbene, and/or v) :CH2.
In one embodiment of the invention, radicals are generated or produced when the composition or at least one constituent of the composition is excited in at least one of the following ways: i) by being exposed to redox reactions, ii) by being subjected to radiation, preferentially ionizing or electromagnetic radiation, iii) by being heated, iv) by being exposed to electrical discharges, v) by undergoing electrolysis, vi) by being exposed to pH variation.
In one embodiment of the invention, the center a free radical production is at least one atom, molecule, chemical function, ionized or not, charged or not, which produces at least one free radical, preferentially in such a way that the quantity or concentration of free radicals present when the composition or at least one constituent of the composition comprises the center of activity or free radical production is more important than the quantity or concentration of free radicals present when the composition or at least one constituent of the composition does not comprise the center of activity or free radical production, where the comparison is preferentially being made using the same or similar conditions of excitation for the composition or at least one constituent of the composition comprising the center of activity or free radical production than for the composition or at least one constituent of the composition not comprising the center of activity or free radical production.
In one embodiment of the invention, the center a free radical capture is at least one atom, molecule, chemical function, ionized or not, charged or not, which captures at least one free radical, preferentially in such a way that the quantity or concentration of free radicals present when the composition or at least one constituent of the composition comprises the center of activity or free radical capture is less important than the quantity or concentration of free radicals present when the composition or at least one constituent of the composition does not comprise the center of activity or free radical capture, where the comparison is preferentially being made using the same or similar conditions of excitation for the composition or at least one constituent of the composition comprising the center of activity or free radical capture than for the composition or at least one constituent of the composition not comprising the center of activity or free radical capture.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are at least one anti-oxidizing or reducing compound.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are at least one oxidizing or anti-reducing compound.
In some cases, the oxidizing compound can be a simple body, a compound, or an ion, that receives or is able to receive at least one electron from another chemical species than the oxidizing compound preferentially during a redox reaction.
In some other cases, the reducing compound can be a simple body, a compound, or an ion, that gives or provides or is able to give or provide at least one electron to another chemical species than the oxidizing compound preferentially during a redox reaction.
In some cases, an anti-oxidizing compound is a compound that prevents oxidation preferentially of the nanoparticle or at least one constituent of the composition or of at least one substance of the composition or at least one constituent of the composition.
In some cases, an oxidizing compound is a compound that favors or triggers oxidation preferentially of the nanoparticle or at least one constituent of the composition or of at least one substance of the composition or at least one constituent of the composition.
In some cases, an anti-reducing compound is a compound that prevents reduction preferentially of the nanoparticle or at least one constituent of the composition or of at least one substance of the composition or at least one constituent of the composition.
In some cases, a reducing compound is a compound that favors or triggers reduction preferentially of the nanoparticle or at least one constituent of the composition or of at least one substance of the composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein at least two nanoparticle or at least one constituent of the compositions in the composition or at least one constituent of the composition arrange in a chain or remained arranged in a chain or comprise at least one center of activity or free radical/production, preferentially at or below 0° C.
In some cases, the at least two nanoparticle or at least one constituent of the compositions can be arranged in chains or comprise at least one center of activity or free radical/production at a temperature that is at or lower than 103, 500, 100, 50, 20, 5, 2, 1, 0, −2, −5, −20, −50, −77, −100, −200 or −273° C.
In some cases, the at least two nanoparticle or at least one constituent of the compositions can be arranged in chains or comprise at least one center of activity or free radical/production at a temperature that is at or larger than −273° C., −200, −100, −77, −50, −20, −5, −2, 0, 2, 5, 10 50 or 100° C.
The invention relates to the composition or at least one constituent of the composition according to the invention, where the at least one nanoparticle or at least one constituent of the composition is in a liquid suspension and the composition or at least one constituent of the composition preferentially has at least one of the following properties:
In one embodiment of the invention, the at least one nanoparticle or at least one constituent of the composition is in liquid suspension when it is mixed with a liquid. In some cases, the liquid can be water, an isotonic liquid, a liquid comprising an excipient, a surfactant, or an oil.
In some cases, the composition or at least one constituent of the composition is isotonic, i.e. it preferentially has:
In another embodiment of the invention, OPBP and/or OPComp is/are larger than 10−10, 10−5, 10−3, 10−1, 0, 1, 5, 10, 50 or 100 bar or atm.
In still another embodiment of the invention, OPBP and/or OPComp is/are smaller than 1010, 105, 103, 10, 0, 10−1, 10−3, 10−5, or 10−10 bar or atm.
In one embodiment, the value of abs(OPBP−OPComp)/OPBP is kept low or the composition or at least one constituent of the composition is maintained isotonic to avoid blood pressure increase, preferentially during or after administration of the composition or at least one constituent of the composition to the body part.
In some cases, the composition or at least one constituent of the composition can be hypertonic, i.e. that preferentially in this case, the composition or at least one constituent of the composition causes at least one cell or body part to shrink.
In some other cases, the composition or at least one constituent of the composition can be hypotonic, i.e. that preferentially in this case, the composition or at least one constituent of the composition causes at least one cell or body part to swell.
In still some other cases, the composition or at least one constituent of the composition can be isotonic, i.e. that preferentially in this case, the composition or at least one constituent of the composition produces no change in cell or body part volume or less change in cell or body part volume than for hypertonic or hypotonic composition or at least one constituent of the composition.
In one embodiment of the invention, the solutes or solute compounds in the composition or at least one constituent of the composition, preferentially the nanoparticle or at least one constituent of the composition, chain of nanoparticie or at least one constituent of the composition and/or cryoprotectant or protectant compound have a concentration, preferentially designated as Csolute, preferentially designated as Csolute,outside outside a cell or outside a body part, preferentially designated as Csolute,inside inside a cell or inside a body part, which is such that abs(Csolute,outside−Csolute,inside)/Csolute,outside is lower than or equal to 100, 50, 25, 10, 5, 2, 1, 0, 10−1, 10−3 or 10−5%.
In some other cases, (Csolute,outside−Csolute,inside)/Csolute,outside is larger than or equal to 10−5, 10−3, 10−1, 0, 1, 2, 5, 10, 25, 50, 75 or 99%.
In one embodiment of the invention, the composition or at least one constituent of the composition is isosmotic or the osmolarity or osmotic pressure of the composition or at least one constituent of the composition, preferentially outside at least one cell or body part is the same as the osmolarity or osmotic pressure of an intracellular medium or body part or composition or at least one constituent of the composition comprised inside at least one cell or body part.
In some cases, the body part can designate a liquid or liquid medium comprised in a body part such as the intracellular medium or plasma or blood.
In some cases, the composition or at least one constituent of the composition is comprised in the body part, preferentially after or with administration to the body part.
In some other cases, the composition or at least one constituent of the composition is comprised outside the body part, preferentially before or without administration to the body part.
In some cases, the composition or at least one constituent of the composition can be hypoosmotic.
In some other cases, the composition or at least one constituent of the composition can be hyperosmotic.
In one embodiment of the invention, the osmolality or osmolarity of the composition or at least one constituent of the composition is larger than or equal to 10−10, 10−5, 10−3, 10−1, 0, 1, 5, 10, 50, 100, 200, 250, 290, 300, 310, 350, 400, 500, 103, 105 or 1010 mOsm/kg or mOsm/L.
In another embodiment of the invention, the osmolality or osmolarity of the composition or at least one constituent of the composition is lower than or equal to 1010, 105, 103, 500, 350, 310, 300, 250, 200, 100, 10, 5, 1, 0, 10−3, 10−5 or 10−10 mOsm/kg or mOsm/L.
In another embodiment of the invention, the osmolality of the composition or at least one constituent of the composition is equal to, close to, or does not differ by more than 1, 5, 10, 50, or 100% from the human plasma osmolality, preferentially comprised between 275 and 299 milli-osmoles per kilogram.
In another embodiment of the invention, the hydrostatic pressure or osmotic pressure or arterial pressure or systolic pressure or diastolic pressure or pressure of the composition or at least one constituent of the composition or in the presence of the composition or at least one constituent of the composition or following the administration of the composition or at least one constituent of the composition or applied by the composition or at least one constituent of the composition on the body part or the force pushing the composition or at least one constituent of the composition or at least one compound or solute of the composition or at least one constituent of the composition preferentially per unit surface of the body part, preferentially in some cases from outside the body part or at least one cell of the body part to inside the body part or inside at least one cell of the body part, preferentially in some other cases from inside the body part or inside at least one cell of the body part to outside the body part or outside at least one cell of the body part, is designated as ξ.
In some cases, ξ can be lower than or equal to 105, 103, 500, 200, 180, 150, 140, 100, 50, 20, 10, 1, 0, 10−1, 10−3 or 10−5 mmHg or PSI or bar or mbar.
In some other cases, ξ be larger than or equal to 10−5, 10−3, 10−1, 0, 1, 5, 10, 100, 120, 150, 500, 103, 105, or 1010 mmHg or PSI or bar or mbar.
In one embodiment of the invention, the volume occupied by water or a liquid and preferentially the cryoprotectant or protectant compound in the composition or at least one constituent of the composition is larger than the volume occupied by the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition. In this case, the at least one nanoparticle or at least one constituent of the composition or chain is preferentially suspended in a liquid, preferentially water, which comprises the cryoprotectant or protectant compound. In this case, the volume of liquid is preferentially maintained sufficiently large or larger than the volume occupied by the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition to enable the suspension of the at least one chain in the composition or at least one constituent of the composition or motion or Brownian motion of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition.
In some cases, the volume occupied by water or liquid and preferentially the cryoprotectant or protectant compound in the composition or at least one constituent of the composition is larger than the volume occupied by the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in liquid form.
In some other cases, the volume occupied by water or liquid and preferentially the cryoprotectant or protectant compound in the composition or at least one constituent of the composition is lower than the volume occupied by the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in powder form.
In some cases, a can be equal to Vliq/Vchain, where Vliq and Vchain are the volumes occupied by the liquid preferentially comprising the cryoprotectant or protectant compound and at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, respectively, where the two volumes can preferentially be measured by separating the at least one nanoparticle or at least one constituent of the composition or chain from the liquid, using for example by magnetic separation or using a magnet to attract the nanoparticle or at least one constituent of the composition or chains and separate them from the liquid.
In another embodiment of the invention, the mass of water or liquid and preferentially cryoprotectant or protectant compound in the composition or at least one constituent of the composition is larger than the mass of at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition. In this case, the at least one nanoparticle or at least one constituent of the composition or chain is preferentially suspended in a liquid, preferentially water, which comprises the cryoprotectant or protectant compound. In this case, the mass of liquid is preferentially maintained sufficiently large or larger than the mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition to enable the suspension of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition or motion or Brownian motion of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition.
In some cases, the mass of water or liquid and preferentially cryoprotectant or protectant compound in the composition or at least one constituent of the composition is larger than the mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in liquid form.
In some other cases, the mass of water or liquid and preferentially cryoprotectant or protectant compound in the composition or at least one constituent of the composition is lower than the mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in powder form.
In some cases, α can be equal to mliq/mchain, where mliq and unchain are the masses of the liquid preferentially comprising the cryoprotectant or protectant compound and at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, respectively, where the two masses can preferentially be measured by separating the at least one nanoparticle or at least one constituent of the composition or chain from the liquid, using for example magnetic separation or using a magnet to attract the nanoparticle or at least one constituent of the composition or chains and separate them from the liquid, where unchain and/or mliq can preferentially be measured by or following evaporation of the liquid from the composition or at least one constituent of the composition or lyophilization or desiccation of the composition or at least one constituent of the composition.
In one embodiment of the invention, the percentage in mass of water or liquid, preferentially comprising the cryoprotectant or protectant compound, preferentially not comprising the at least one nanoparticle or at least one constituent of the composition or chain, in the composition or at least one constituent of the composition, is larger than the percentage in mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition.
In some cases, the percentage in mass of water or liquid preferentially comprising the cryoprotectant or protectant compound, preferentially not comprising the at least one nanoparticle or at least one constituent of the composition or chain, in the composition or at least one constituent of the composition is larger than the mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in liquid form.
In one embodiment of the invention, the percentage in mass of water or liquid, preferentially comprising the cryoprotectant or protectant compound, preferentially not comprising the at least one nanoparticle or at least one constituent of the composition or chain, in the composition or at least one constituent of the composition, is smaller than the percentage in mass of the at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition by a factor α of at least 1.1, 2, 5, 10 or 103. In this case, the composition or at least one constituent of the composition is preferentially in powder form.
In some cases, a can be equal to Pliq/Pchain, where Pliq and Pchain are the percentages in masses of the liquid preferentially comprising the cryoprotectant or protectant compound and at least one nanoparticle or at least one constituent of the composition or chain in the composition or at least one constituent of the composition, respectively, where the two percentages in masses can preferentially be measured by separating the at least one nanoparticle or at least one constituent of the composition or chain from the liquid, using for example magnetic separation or using a magnet to attract the nanoparticle or at least one constituent of the composition or chains and separate them from the liquid, where Pchain and/or Pliq can preferentially be measured by or following evaporation of the liquid from the composition or at least one constituent of the composition or lyophilization or desiccation of the composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, where the at least one nanoparticle or at least one constituent of the composition or chain is in powder form, and the composition or at least one constituent of the composition has preferentially at least one of the following properties:
In some cases, the composition or at least one constituent of the composition is lyophilized, desiccated, dried or dehydrated when water or liquid is removed, preferentially partly or fully, from the composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the composition or at least one constituent of the composition, in which the composition or at least one constituent of the composition is lyophilized or desiccated or dried or dehydration or treated by lyophilization or desiccation or water removal or liquid removal or dehydration.
In one embodiment of the invention, the composition or at least one constituent of the composition is dehydrated or water or liquid is removed or absent from the composition or at least one constituent of the composition. In this case, the percentage in mass of water or liquid in the composition or at least one constituent of the composition is preferentially larger before the composition or at least one constituent of the composition is dehydrated or before water or liquid is removed from the composition or at least one constituent of the composition than after the composition or at least one constituent of the composition is dehydrated or after water or liquid is removed from the composition or at least one constituent of the composition. In this case, the volume of water or liquid occupied in the composition or at least one constituent of the composition is preferentially larger before the composition or at least one constituent of the composition is dehydrated or before water or liquid is removed from the composition or at least one constituent of the composition than after the composition or at least one constituent of the composition is dehydrated or after water or liquid is removed from the composition or at least one constituent of the composition. In this case, water or liquid is removed from the composition or at least one constituent of the composition without removing the at least one nanoparticle or at least one constituent of the composition or chain and preferentially also the cryoprotectant or protectant compound from the composition or at least one constituent of the composition.
In one embodiment of the invention, the percentage in mass of water in the composition or at least one constituent of the composition is larger than 10−3, 10−1, 0, 1, 5, 10, 25, 50, 80 or 99%, preferentially before water is or has been removed from the composition or at least one constituent of the composition.
In another embodiment of the invention, the percentage in mass of water in the composition or at least one constituent of the composition is lower than or equal to 100, 99, 80, 50, 25, 10, 5, 2, 1 or 0%, preferentially after water is or has been removed from the composition or at least one constituent of the composition.
In another embodiment of the invention, the composition or at least one constituent of the composition is lyophilized or the composition or at least one constituent of the composition is first cooled down, preferentially below or at 103, 100, 50, 10, 0, −10, −50, −77, −200, −273° C., and second water or liquid is removed from the composition or at least one constituent of the composition, preferentially without removing the at nanoparticle or at least one constituent of the composition or least one nanoparticle or at least one constituent of the composition or chain and/or cryoprotectant or protectant compound from the composition or at least one constituent of the composition.
In another embodiment of the invention, the composition or at least one constituent of the composition is lyophilized, dehydrated, dried and/or desiccated, and water or liquid is preferentially removed from the composition or at least one constituent of the composition preferentially without removing the at least one nanoparticle or at least one constituent of the composition or chain and/or cryoprotectant or protectant compound from the composition or at least one constituent of the composition.
In one embodiment, the composition or at least one constituent of the composition is comprised in a tablet or is a tablet form or powder form, preferentially after water has been removed from the composition or at least one constituent of the composition.
In one embodiment, the composition or at least one constituent of the composition is stoked or stored or unused, preferentially for more than 0, 1, 10, 102, 103, 105, or 1010 hour(s), day(s), month(s), year(s), preferentially without being used or administered to a body part, preferentially in powder form, preferentially without losing at least one its property or without losing its nanoparticle or at least one constituent of the composition coating or nanoparticle or at least one constituent of the composition core, partly or fully.
In one embodiment of the invention, the moisture or water content of the composition or at least one constituent of the composition is lower than or equal to 100, 75, 50, 25, 10, 5, 4, 2, 1 or 0% w/w, preferentially weight of water for weight of composition or at least one constituent of the composition or weight of water by weight of composition or at least one constituent of the composition or mass of water for mass of composition or at least one constituent of the composition or mass of water by mass of composition or at least one constituent of the composition, preferentially after water has been removed from the composition or at least one constituent of the composition.
In another embodiment of the invention, the moisture or water content of the composition or at least one constituent of the composition is larger than or equal to 0, 1, 2, 4, 5, 10, 25, 50, 75 or 100% w/w, preferentially weight of water for weight of composition or at least one constituent of the composition or weight of water by weight of composition or at least one constituent of the composition or mass of water for mass of composition or at least one constituent of the composition or mass of water by mass of composition or at least one constituent of the composition, preferentially before water has been removed from the composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, where the at least one nanoparticle or at least one constituent of the composition or chain and preferentially the cryoprotectant or protectant compound, which is/are preferentially in powder form, is/are suspended or resuspended in a liquid or water or treated by suspension or resuspension in a liquid or water, preferentially more than 1, 2, 5 or 10 times. In this case, water is preferentially removed from the composition or at least one constituent of the composition before the composition or at least one constituent of the composition is suspended or resuspended in a liquid or water.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or at least one constituent of the composition or at least one of its chemical functions or atoms preferentially forms a first type of interaction or chemical bond with an atom or chemical function of the core, wherein the cryoprotectant or protectant compound or at least one of its chemical functions or atoms preferentially forms a second type of interaction or chemical bond with the coating or at least one of its chemical functions or atoms,
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or at least one constituent of the composition is selected in the group of compounds consisting of: 1) citric acid, 2) oleic acid, 3) polymethacrylic acid, 4) poly(ethyleneoxide)-b-poly(methacrylic) acid), 5) polyacrylic acid (PAA), 6) polylactic acid, 7) poly(ethylene oxide)-blockpoly(glutamic) acid, 8) Phosphonic acid 9) Albumin, 10) Alendronate, 11) Alginate, 12) Au, Al2O3, 13) Aluminium, 14) Aluminium hydroxide, 15) Arabinogalactan, 16) Bentonite, 17) Carboxymethylcellulose, 18) Cellulose, 19) Chitosan, 20) Cholesterol, 21) Citrate, 22) Dextran, 23) Dimercaptosuccinic acid, 24) Dopamine, 25) DOPC or Dioleoylphosphatidylcholine or phospholipd, 26) DTAP or Di(tert.-amyl)peroxide, 27) DVB or Divinylbenzene, 28) Ethylcellulose, 29) Erythrocyte, 30) at least one fatty acids, 31) Ferrite, 32) Folic acid, 33) Gelatin, 34) serum albumin, preferentially human, 35) Liposome, 36) MIPS or Inositol-3-phosphate synthase, 37) MnO or manganese oxide, 38) Mn3O4, 39) Oleic acid, 40) at least one polymer or enantiomer, 41) PEI or Polyetherimide, 42) PEG or Polyethylene glycol, 43) Poly(ethylene oxide) or PEO, 44) PGA or Polyglycolic acid, 45) PLA or poly(lactide acid), 46) PLGA or PLG or poly(lactic-co-glycolic acid), 47) Phosphatidylcholine, 48) Phosphorylcholine, 49) Pluronic, 50) Polyacrylamide, 51) Polyacrylic acid or PAA, 52) Polyaniline, 53) Polyethylene glycol with/without terminal carboxyl groups, 54) Peptide or Polypeptide, 55) Poly(vinyl alcohol) or PVA, 56) Ploy(N-isopropylacrylamide) or PIA, 57) Poly(vinylpyrrolidone) or PVP, 58) Poly(oligoethylene oxide) or POO, 59) Poly(N,N-dimethyl ethylamino acrylate, 60) Poly(imine), 61) Poly(acrylic acid), 62) Poly-D-L lactide, 63) Polyalkylcyanoacrylate, 64) Polymer such as PAMAM or Poly(amidoamine) or PDMAEMA or poly(2-(dimethylamino)ethyl methacrylate) or PPEGMA or Poly (ethylene glycol) methyl ether methacrylate, 65) Poly NIPAAM or Poly (N-isopropylacrylamide) or temperature-responsive polymer or temperature-sensitive polymer 66) Polyacrylic acid, 67) Polydipyrrole or dicarbazole, 68) Poly-L-lysine, 69) Polymethylmethaacrylate, 70) Polymersome, 71) Polystyrene, 72) PVA or Polyvinyl Alcohol, 73) PVP or Polyvinylpyrrolidone, 74) Silica, preferentially amorphous or mesoporous, 75) Silane, 76) SiO2, 77) Sodium Oleate, 78) Starch, 79) styrene, preferentially styrene-divinylbenzene, 80) TaOx, 81) ZrO2, 82) at least one metal or semi-metal, 83) at least one metal oxide or semi-metal oxide, 84) at least one alkali metal, 85) at least one alkaline earth metal, 86) at least one transition metal, 87) at least one post-transition metal, 88) at least one metalloid, 89) at least one lanthanide, 90) at least one actinide, 91) at least one non-metal, 92) at least one halogen, 93) at least one noble gas, and 94) any derivative or combination of any of these compounds.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or at least one constituent of the composition is selected in the group of family of compounds consisting of: i) Polysacharides such as Agarose, alginate, carregeenan, chitosan, dextran, haparin, Gum Arabic, Pullulan and Starch, ii) Acids such as citric acids, oleic acids, polymethacrylic acid, poly(ethyleneoxide)-b-poly(methacrylic acid, polyacrylic or PAA acid, polylactic acid, poly(ethylene oxide)-blockpoly(glutamic acid, Phosphonic acid, Dimercaptosuccinic acid, fatty acid, folic acid, oleic acid, poly(lactide acid or PLA, PAA or Polyacrylic acid, compounds comprising at least one carboxylic function, iii) Polymers such as Dextran, Poly(ethylene oxide), Poly(vinyl alcohol), Ploy(N-isopropylacrylamide), Poly(vinylpyrrolidone), Poly(oligoethylene oxide), Poly(N,N-dimethyl ethylamino acrylate), Poly(imine), Poly(acrylic acid), iv) Carboxylates, v) inorganic compounds such as SiO2, Al2O3, ZrO2, ferrites, MnO, Mn3O4, Au, Bentonite, Carbon such as inactivated, activated, graphitized carbon, vi) at least one metals, vii) organic compounds such as MIP, Cellulose, DVB, Ppy, Chitosan, Polyacrylamide, alginate, PEI, surfactants, viii) compounds comprising Phosphate, ix) compounds comprising Silica, x) compounds comprising Gold, xi) compounds comprising Dextran based, xii) compounds comprising PEG, xiii) compounds comprising PVA, xiv) compounds comprising Alginate, xv) compounds comprising Chitosan, xvi) compounds comprising the chemical function Alcohol, xvii) compounds comprising the chemical function Amide, and xviii) compounds comprising the chemical function Aldehyde.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or at least one constituent of the composition has at least one chemical group selected in the group of i) OH−, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acid derivative of at least one of these compounds.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or coating material or at least one constituent of the composition comprises a chemical function, preferentially selected in the group consisting of i) OH—, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acid derivative of at least one of these functions, which has an interaction or forms a chemical bond with an atom or chemical group located at the surface of the core, preferentially a hydroxyl group (Fe—OH), where the interaction or bond is preferentially selected in the group consisting of: i) an electrostatic interaction or bond, i.e. preferentially due to the difference in charge between the coating and the surface or core of the core, ii) a hydrophobic interaction or bond, iii) a Chelating interaction or bond, iv) a metallic interaction or bond, v) a Covalent interaction or bond.
In one embodiment of the invention, the cryoprotectant or protectant compound interacts or forms with the core or coating of the nanoparticle or at least one constituent of the composition hydrogen or van der walls or London bonds or interaction, preferentially when water molecules are displaced or cooled down.
In one embodiment of the invention, as the cryoprotectant or protectant compound replaces the water molecules, preferentially when the composition or at least one constituent of the composition is cooled down, the coating and/or core of the nanoparticle or at least one constituent of the composition preferentially retains its structure and function and/or at least two nanoparticles or at least two constituents of the composition remain arranged in chain and/or the at least one center of activity or free radical capture/production remains comprised in the nanoparticle or at least one constituent of the composition.
In one embodiment of the invention, the cryoprotectant or protectant compound is not used to protect the at least one nanoparticle or at least one constituent of the composition or at least one chain from the denaturation and/or destruction and/or loss of primary, secondary, tertiary or quaternary structure of DNA, RNA, protein(s), lipid(s), enzyme(s), or at least one biological molecule.
In one embodiment of the invention, the cryoprotectant or protectant compound prevents a change, decrease, increase of the size or of at least one nanoparticle or at least one constituent of the composition or chain by more than 100, 50, 10 or 1%, where this percentage is preferentially equal to S2-S1/S1 or P2-P1/P1, where S1, P1, S2 and P2 are preferentially the sizes and properties of at least one nanoparticle or at least one constituent of the composition or chain before (S1, P1) and after (S2, P2) cooling down the at least one nanoparticle or at least one constituent of the composition or chain preferentially by more than 1, 50 or 100° C., preferentially from an initial temperature, preferentially above −273, −100, −50, 0, 5, or 10° C., preferentially down to a final temperature, preferentially lower than 100, 50, 0 or −50° C.
In some cases, the property of at least one nanoparticle or at least one constituent of the composition or chain can be: i) the percentage in mass of at least one metal in the at least one nanoparticle or at least one constituent of the composition or chain, ii) the number of nanoparticle or at least one constituent of the composition in the chain, iii) the surface charge of the at least one nanoparticle or at least one constituent of the composition or chain, iii) the percentage of water in the at least one nanoparticle or at least one constituent of the composition or chain, iv) the coercivity or remanent magnetization of the at least one nanoparticle or at least one constituent of the composition or chain, v) the core diameter of the at least one nanoparticle or at least one constituent of the composition, vi) the coating thickness of the at least one nanoparticle or at least one constituent of the composition, vii) the isotonicity or osmolality of the at least one nanoparticle or at least one constituent of the composition or chain.
In one embodiment, the composition or at least one constituent of the composition comprising the cryoprotectant or protectant compound does not comprise ice, while the composition or at least one constituent of the composition not comprising the cryoprotectant or protectant compound comprises ice or the composition or at least one constituent of the composition comprising the cryoprotectant or protectant compound comprises more ice than the composition or at least one constituent of the composition not comprising the cryoprotectant or protectant compound, preferentially when the composition or at least one constituent of the composition is cooled down and/or he lyophilized,
In one embodiment of the invention, the composition or at least one constituent of the composition comprising the cryoprotectant or protectant compound has a lower melting point than the composition or at least one constituent of the composition not comprising the cryoprotectant or protectant compound.
In one embodiment of the invention, the cryoprotectant or protectant compound is a nonpermeable cryoprotectant or protectant compound.
In some cases, the cryoprotectant or protectant compound can be selected among: a sugar, trehalose, sucrose, starch, hydroxyethyl starch, polyvinyl pyrrolidone, and/or polyethylene oxide.
Preferentially, a nonpermeable cryoprotectant or protectant compound does not enter cells or coatings and thus preferentially stays extracellular or outside the coating when the composition or at least one constituent of the composition is cooled down.
In some cases, the composition or at least one constituent of the composition is cooled during or for cryopreservation of the composition or at least one constituent of the composition.
Preferentially, a nonpermeable cryoprotectant or protectant compound is used to protect cells during the cooling of the composition or at least one constituent of the composition.
Preferentially, when a cooling or slow cooling or slow freezing rate is applied to the composition or at least one constituent of the composition or the composition or at least one constituent of the composition is cooled down, preferentially slowly, there is sufficient time for water or liquid comprised in the nanoparticie or at least one constituent of the composition preferentially in the coating of the nanoparticle or at least one constituent of the composition to move out of the coating, preferentially under the osmotic pressure, preferentially resulting in a reduction in the volume of the coating. In the absence of a cryoprotectant or protectant compound, such behavior can destroy or damage the coating and/or result in the destruction of the nanoparticle or at least one constituent of the composition chain arrangement or of the center of activity or free radical/production.
In one embodiment, a cryoprotectant or protectant compound, preferentially a permeable cryoprotectant or protectant compound can enter a cell or the coating and preferentially preserve the cell or coating, preferentially from the osmotic pressure or damage or destruction.
In some cases, the cryoprotectant or protectant compound can be dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, or propylene glycol, in some cases, the cryoprotectant or protectant compound can be used to induce the vitrification of intracellular environment or of the coating or of the inner part or surface part of the coating or nanoparticle or at least one constituent of the composition or core, preferentially before ice crystal formation, preferentially in the composition or at least one constituent of the composition, preferentially preventing excessive loss volume of cell or coating or nanoparticie or at least one constituent of the composition or core.
In one embodiment of the invention, the cryoprotectant or protectant compound is selected in the group consisting of: i) natural product, ii) soybean flour product, iii) carbohydrate, iv) lipid, v) saccharide, vi) zwitterionic molecule, vii) l-carnitine, and viii) antifreeze compounds such as antifreeze proteins.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the coating or at least one constituent of the composition or at least one of its chemical functions or atoms forms a first type of interaction or chemical bond with an atom or chemical function of the core, wherein the cryoprotectant or protectant compound or at least one of its chemical functions or atoms forms a second type of interaction or chemical bond with the coating or at least one of its chemical functions or atoms,
In some cases, the coating or coating material or at least one constituent of the composition can comprise a chemical function, preferentially selected in the group consisting of i) OH−, ii) NH2, iii) COOH, iv) Thiol, v) Phosphate, and a basic or acid derivative of at least one of these functions.
In some other cases, the core or at least one constituent of the composition can comprise a chemical function, preferentially located at the surface of the core of the nanoparticle or at least one constituent of the composition, preferentially a hydroxyl group (Fe—OH).
The invention also relates to the composition or at least one constituent of the composition according to the composition or at least one constituent of the composition, wherein the cryoprotectant or protectant compound or at least one constituent of the composition is selected in the group consisting of: 1) Acetamide, 2) Acetate, 3) Albumin, 4) Amino acids, 5) Ammonium acetate, 6) Arginine, 7) Alcohols containing at least one or two hydroxyl groups, 8) Bridger, 9) Choline magnesium chloride sodium bromide, 10) Diethyl glycol, 11) Dimethylacetamide, 12) Dimethyl sulfoxide (DMSO), 13) Disaccharide, 14) Erythritol, 15) Ethanol, 16) Ethylene glycol, 17) Formamide, 18) Fructose, 19) Glucose, 20) Glycerol, 21) Glycerol 3-phosphate, 22) Glycol such as Diethyl glycol or Triethylene glycol, 23) Glycine, 24) Lactose, 25) L-tyrosine, 26) lysine hydrochloride, 27) Mannitol, 28) MDP (2-Methyl-2,4-pentanediol), 29) Phenylalanine, 30) Planic, 31) Polymers, 32) Polyethylene glycol such as PEG4000, polyethylene glycol succinate, folate modified distearoylphosphatidyl ethanolamine-polyethylene glycol, 33) Polyethyleneimine (PEI), 34) Polyvinylpyrrolidone (PVP), 35) Proline, 36) Propylene glycol, 37) Protein, 38) Pyridine (Pyridine-N-Oxide), 39) Ribose, 40) Sarcosine, 41) Serine, 42) Serum albumin, 43) Sodium bromide, 44) Sodium chloride, 45) Sodium dodecyl sulfonate, 46) Sodium glutamate, 47) Sodium iodide, 48) Sodium sulfate, 49) Sorbitol, 50) Starch (hydroxyethyl starch), 51) Sugar, 52) Sucrose, 53) The cell bank series, 54) Trehalose, 55) Triethylene glycol, 56) Trimethylamine, 57) Tween 80, 58) Tryptophan, 59) Valine, 60) Xylose, and 61) a combination or derivative of any of these compounds.
In one embodiment of the invention, the composition or at least one constituent of the composition is desiccated or lyophilized or dehydrated or dried. In this case, the percentage in mass of water or liquid in the composition or at least one constituent of the composition is preferentially smaller than 100, 75, 50, 75, 20, 10, 5, 2, 1, 0, 10−1, 10−3 or 10−5%. In this case, the quantity of water or liquid in the composition or at least one constituent of the composition is preferentially smaller than 100, 50, 10, 5, 2, 1, 10−1, 10−3, or 10−5 grams of water or liquid in the composition or at least one constituent of the composition per gram of composition or at least one constituent of the composition.
In some other cases, the composition or at least one constituent of the composition can be non-desiccated, non-dehydrated, or non-dehydrated or dried. In this case, the percentage in mass of water or liquid in the composition or at least one constituent of the composition is preferentially larger than 10−5, 10−3, 10−1, 0, 1, 5, 10, 25, 50, 75 or 100%. In this case, the quantity of water or liquid in the composition or at least one constituent of the composition is preferentially larger than 10−10, 10−5, 10−1, 0, 1, 5, 10 or 50, 10 grams of water or liquid in the composition or at least one constituent of the composition per gram of composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, which is lyophilized or desiccated or dehydrated, wherein the percentage in mass of cryoprotectant or protectant compound is preferentially larger than the percentage in mass of coating in the lyophilized or desiccated or dehydrated composition or at least one constituent of the composition, wherein the lyophilized or desiccated or dehydrated composition or at least one constituent of the composition is preferentially more stable than non-lyophilized or non-desiccated or non-dehydrated composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition, where the percentage in mass of the cryoprotectant or protectant compound is comprised between 0.5 and 50%.
In some cases, the percentage in mass of a substance comprised in the composition or at least one constituent of the composition, e.g. the cryoprotectant or protectant compound, nanoparticle or at least one constituent of the composition core, nanoparticle or at least one constituent of the composition coating, chain, liquid or water, is equal to or proportional to the mass of this substance divided by the mass of all the substances comprised in the composition or at least one constituent of the composition.
In some cases, the percentage in mass of the cryoprotectant or protectant compound, nanoparticle or at least one constituent of the composition coating, nanoparticle or at least one constituent of the composition core, at least one chain, liquid or water, in the composition or at least one constituent of the composition is larger than 10−5, 10−3, 10−1, 0, 1, 5, 10, 25, 50, 75, 90 or 99%.
In some other cases, the percentage in mass of the cryoprotectant or protectant compound, nanoparticle or at least one constituent of the composition coating, nanoparticle or at least one constituent of the composition core, at least one chain, liquid or water, in the composition or at least one constituent of the composition is lower than 100, 75, 50, 25, 10, 5, 2, 1, 0, 10−3 or 10−5%.
In one embodiment of the invention, the percentage in mass of cryoprotectant or protectant compound in the composition or at least one constituent of the composition according to the invention is larger than the percentage in mass of nanoparticle or at least one other constituent of the composition, coating, nanoparticle core, at least one chain, liquid and/or water, in the composition or at least one constituent of the composition.
In another embodiment of the invention, the percentage in mass of cryoprotectant or protectant compound in the composition or at least one constituent of the composition according to the invention is smaller than the percentage in mass of nanoparticle or at least one constituent of the composition coating, nanoparticle or at least one constituent of the composition core, at least one chain, liquid and/or water, in the composition or at least one constituent of the composition.
In one embodiment of the invention, the core of the nanoparticle or at least one constituent of the composition in the composition or at least one constituent of the composition according to the invention has at least one property selected in the group consisting of: a) it is ferrimagnetic, b) it is composed of maghemite or magnetite or of an intermediate composition or at least one constituent of the composition between maghemite and magnetite, c) it has a size comprised between 0.1 and 100 nm, and d) it comprises at least one crystallographic plane.
In another embodiment of the invention, the coating of the nanoparticle or at least one constituent of the composition in the composition or at least one constituent of the composition according to the invention has at least one property selected in the group consisting of: a) it has a thickness lower than 10 μm, b) it comprises a number of crystallographic planes per unit surface that is lower than the number of crystallographic planes per unit surface of the core, c) it has a non-neutral charge, d) it is amorphous and e) it is organic, and f) it has a thickness that is lower than 10 nm or than the diameter of the core.
In another embodiment of the invention, the cryoprotectant or protectant compound in the composition or at least one constituent of the composition according to the invention is comprised in a matrix or volume embedding the core and/or coating of the nanoparticle or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the core or nanoparticle or at least one constituent of the composition core is synthesized by a living organism or nanoparticle or at least one constituent of the composition producing cells, preferentially a magnetotactic bacterium, while preferentially the coating is not synthesized by the nanoparticle-producing cell.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized biologically or by a living organism, designated as synthetizing living organism, which preferentially consists or comprises at least 1, 2, 5, 10, 103, 106 or 109 eukaryotic cell(s), prokaryotic cell(s), or part of these cells. In some cases, part of eukaryotic or prokaryotic cell(s) can be biological material originating or produced by these cells such as RNA, DNA, organelle, nucleolus, nucleus, ribosome, vesicle, rough endoplasmic reticulum, Golgi apparatus, cytoskeleton, smooth endoplasmic reticulum, mitochondrion, vacuole, cytosol, lysosome, centrosome, cell membrane. In some cases, a biological synthesis can be defined as a synthesis involving a majority of steps, or more than 1, 2, 5 or 10 steps, or more than 1, 2, 5, 25, 50, 75 or 90% of steps, which involve chemical reactions occurring with the involvement of at least 1, 2, 10, 103, 106 or 109 living organisms, or parts of living organisms such as DNA, RNA, proteins, enzymes, lipids.
In some cases, the synthetizing living organism can be magnetotactic bacteria, other types bacteria than magnetotactic bacteria or enzymes of certain bacteria, preferentially synthetizing nanoparticle or at least one constituent of the compositions extra-cellularly, such as Mycobacterium paratuberculosis, Shewanella oneidensi, Geothrix fermentans, ants, fungi, or various plants.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized or produced or crystallized or assembled or transformed into a nanoparticle or at least one constituent of the composition by a compartment, organelle, or other biological material, such as protein, lipid, enzyme, DNA, or RNA, which is preferentially produced by or originates from an eukaryotic or prokaryotic cell.
In another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized by or in at least one eukaryotic cell, prokaryotic cell, or part of this cell.
In another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized by or in: i) the matrix or medium or environment located outside of at least one eukaryotic cell, prokaryotic cell, or part of this cell, or ii) the extracellular matrix.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized by a living organism when at least 1, 2, 5, 10 or 100 step(s) of their production, such as crystallization of iron oxide, stabilization of the iron oxide mineral, organization of the nanoparticle or at least one constituent of the compositions, for example in chains or aggregates, involves or is due to a living organism.
The invention also relates to the nanoparticle or at least one constituent of the compositions for use, wherein the nanoparticle or at least one constituent of the compositions are magnetosomes synthesized by, originating from, extracted from, or isolated from magnetotactic bacteria.
In one embodiment of the invention, the magnetosome is synthesized by, produced by, originates from, extracted from, isolated from magnetotactic bacteria.
In one embodiment of the invention, magnetotactic bacteria are selected from the group consisting of: Magnetospirillum magneticum strain AMB-1, magnetotactic coccus strain MC-1, three facultative anaerobic vibrios strains MV-1, MV-2 and MV-4, the Magnetospirillum magnetotacticum strain MS-1, the Magnetospirillum gryphiswaldense strain MSR-1, a facultative anaerobic magnetotactic spirillum, Magnetospirillum magneticum strain MGT-1, and an obligate anaerobe, and Desulfovibrio magneticus RS-1.
In one embodiment of the invention, a magnetotactic bacterium is defined as a bacterium able to synthesize magnetosomes, wherein these magnetosomes are preferentially characterized by at least one of the following properties: i) they are produced intracellularly, ii) they are magnetic, iii) they comprise a mineral, iv) their core is preferentially composed of a metallic oxide such as iron oxide, v) their core is surrounded by biological material such as lipids, proteins, endotoxins, which can preferentially be removed, vi) they are arranged in chains, vii) they produce heat under the application of an alternating magnetic field.
In one embodiment of the invention, the magnetosomes possess one or several property(ies) in common with the nanoparticle or at least one constituent of the compositions such as at least one magnetic, size, composition or at least one constituent of the composition, chain arrangement, charge, core, mineral, coating, or crystallinity property.
In one embodiment of the invention, magnetosomes comprise the mineral part synthesized by magnetotactic bacteria, i.e. preferentially the crystallized iron oxide produced by these bacteria. In this case, magnetosomes or magnetosome mineral parts preferentially do not comprise proteins, lipids, endotoxins, or biological materials comprising carbon or do not comprise more or comprise less than 0.1, 1, 10, 30, 50 or 75% or percent in mass of carbon, which is/are produced by these bacteria.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are is/are photosensitizer(s), preferentially selected in the group consisting of: 1) Acridine, such as Acridine Orange, acridine yellow, 2) ALA (5-Aminolevulinic acid), 3) Aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) Aminolevulinic acid, delta-Aminolevulinic acid, 5) Antihistamines, 6) Azulene, 7) Bavteriochlorin, 8) TOOKAD or TOOKAD Soluble, 9) WST-11, 10) LUZ11, 11) BC19, 12) BC21, 13) porphyrin such as Benzoporphyrin derivative monoacid ring A (BPD-MA), 14) Chlorin such as Chlorin e6, m-tetrahydroxyphenylchlorin 15) Foscan, 16) Verteporfin, 17) benzoporphyrin derivative mono acid ring A, 18) Monoaspartyl chlorin(e6), 19) talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrin, 23) Chlorine e6 porphrin, 24) Coal Tar and Derivatives, 25) Contraceptives, Oral and Estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview, 29) Dyes such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squaraines, 33) BODIPY dyes, 34) Phenalenones, 35) benzophenoxazinium dyes, 36) Erythrosine, 37) Flavins, 38) Foscan, 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH), 45) Hypericin, 46) Hypocrellin, 47) ICG (Indocyanine Green), 48) Levulan, 49) MAL-methyl aminolevulinate), 50) Meta-tetra(hydroxyphenyl)chlorin (m-THPC), 51) Metvix, 52) Methylene Blue, 53) Monoterpene, 54) Motexafin lutetium (Lu-Tex), 54) N-aspartyl chlorin e6 (NPe6), 55) Nanoparticle or at least one constituent of the composition or nanomaterial, 56) Natural products or compounds, 57) Non-Steroidal Anti-Inflammatory Drugs, 58) Palladium bacteriopheophorbide (WST09), 59) Phatalocyanin dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phthalocyanine such as Liposomal ZnPC, 65) Chloroaluminium sulfonated phthalocyanine (CASP), 66) Silicon phthalocyanine (PC4), 67) RLP068, 68) Porfimer sodium, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-Tetrakis(1-methylpyridinium-4-yl) porphyrin tosylate, 70) XF70, 71) Protoporphyrin, 72) ALA-induced protoporphyrin IX, 73) Psoralens, 74) Quantum dots, 75) Quinones, 76) Riboflavin, 77) Rose Bengal, 78) silicon or Silicon phthalocyanine (Pc4), 79) Sulfonamides, 80) Sulfonylureas, 81) Talaporfin or Talaporfin soudium, 82) Temoporfin, 82) Tetrahydropyrroles, 83) Tin ethyl etiopurpurin, 84) Titanium dioxide, 85) Toldudine blue O, 86) Transition metal compounds such as Ruthenium(II), polypyridyl complexes, ruthenium, rhodium, cyclometalated, Rh(II)-Rh(II) bridged dimer compounds, platinum(II), gold(III), 87) Verteporfin, 88) Vulnic based compound such as Aminovulinic, aminovulinic acid, 89) WST11, and 90) Xanthene.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are is/are sonosensitizer(s), preferentially selected in the group consisting of: 1) ABS-FA, 2) Acrylonitrile Butadiene Styrene, 3) Styrene, 4) Folic acid, 5) AIMP NP, aminoacyl tRNA synthetase complex-interacting multifunctional protein, 6) Au Nanomaterial, 7) gold, 8) Au—MnO nanomaterial, 9) manganese oxide, 10) Antineoplastic drugs, 11) NSAIDs, 12) nonsteroidal anti-inflammatory drug, 13) Artemether, 14) 5-ALA (5-aminolevulinic acid), 15) Acridine, Acridine Orange, 16) Au-doped TiO2, 17) Carbon based nanomaterial, 18) carbon nanotube, 19) Chlorine, 20) Ce6, 21) PTX, Paclitaxel, 22) chemotherapeutic drug or compound, 23) infrared dye or IR783, 24) Curcumin, 25) Cyanine or Cu-Cyanine, 26) DHMS, 27) dimethylsulfure, 28) Docetaxel, 29) chemotherapeutic drug or compound, 30) DOX/Mn-TPPS @RBCS, 31) doxorubicin, 32) manganese, 33) blood cell, 34) red blood cell, cell, 35) polymer, 36) elastomer, 37) Erythosin or Erythosin B, 38) FA or FA-OI or FA-OI NP or folic acid, 39) F3-PLGA@MB/Gd NPs, 40) poly(lactic-co-glycolic acid), 41) gadolinium, 42) Fe—TiO2 or titanium oxide, 43) Fe-VS2, 44) iron, 45) vanadium disulfide, 46) FMSNs-DOX, 47) silica, 48) HCQ, 49) hydrochloroquine, 50) HP, 51) hematoporphyrin, 52) HMME, 53) hematoporphyrin monomethyl ether, 54) HSYA or Hydroxysafflor yellow A, 55) Hypocrellin, Hypocrellin B, 56) IR780, 57) Levofloxacin, 58) LIP3 or Lithium phosphide, 59) Lithium, 60) Liposome or Liposomal nanomaterial, 61) Lomefoxacin, 62) MG@P NPs, 63) MnP or Manganese peroxidase, 64) MnTTP-HSAs, 65) HSA-wrapped metal-porphyrin complex, 66) albumin, 67) MnWOx, 68) MnWOx-PEG, 69), PEG, 70) bimetallic oxide, 71) Mn (III)-HFs, 72) managense, hemoporfin, 73) Nanoroads, 74) Noble metal nanomaterial, 75) OI NP or oxygen indyocyanine, 76) Phthalocyanines, 77) PIO or Pioglitazone, 78) Polymeric nanomaterial, 79) Porphyrin, 80) Pt-doped TiO2, 81) R837, 82) Rose Bengal, 83) Sparfloxacin, 84) TAPP or 5,10,15,20-tetrakis (4-aminophenyl) porphyrin, 85) TiO2 or titanium dioxide nanomaterial, 86) TCPP, isomer, or Tris(1-chloro-2-propyl) phosphate 87) TPI or Thermoplastic Polyimide or thermoplastic polymer, 88) TPZ or Tirapazamine, 89) Transition metal oxide, 90) nanoparticle or at least one constituent of the composition or Janus nanoparticle or at least one constituent of the composition, and 91) Xanthones.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein C1FRPC and/or C2FRPC is/are radio-sensitizer, preferentially selected in the group consisting of: 1) AMG102, 2) AQ4N, 3) Apaziquone (E09), 4) Bromodeoxyuridine, 5) Carbogen, 6) Cetuximab, 7) Chemotherapeutic drug or compound, 8) Chlorpromazine, 9) C-reactive peptide, 10) Curcumin, 11) Diamide, 12) Diethylmaeate, 13) Dihydroartemisinin, 14) Docetaxel, 15) ECI301, 16) Etanidazole, 17) Fludarabine, 18) 5-Fluorouracil, 19) Fluorodeoxyuridine, 20) Gadolynium, 21) Gemcitabine, 22) HER-3 ADC, 23) HSP, 24) Hydrogen peroxide, 25) Hydroxyurea, 26) Hyperbaric oxygen, 27) Hyperthermia, 28) Hypoxic cell cytotoxic agent, 29) Irinotecan, 30) lanthanide-doped radiosensitizer-based metal-phenolic network, 31) Lidocaine, 32) Lododeoxyuridine, 33) Metronidazole, 34) misonidazole, 35) etanidazole, 36) nimorazole, 37) N-Ethylmalemide, 38) malmeide, 39) ethylmalmeide, 40) Nanomaterial such as those consisting of or composed of at least partly or fully gold, silver, bismuth, gadolinium, polysiloxane matrix and gadolinium chelates, hafnium, Tantalum, Zinc, Gadolinium, Germanium, Chromium, Praseodymium, Silicon, iron, platinum, cobalt, manganese, magnesium, iron, Titanium, carbon nanotube, quantum dot, nanoroad, Triflate, or metal oxide, 41) Nelfinavir, 42) Nicotinamide, 43) Nimotuzumab, 44) RNA, or miRNA, or miR-201, or miR-205, or miR-144-5p, or miR-146a-5p, or miR-150, or miR-99a, or miR-139-5p, or miR-320a, 45) Membrane active agent, 46) Mitomycin-C or Mitomycin, 47) Motexafin, 48) NBTXR3, 49) Oligonucleotide, 50) Paclitaxel, 51) Papaverine or Papaverine hydrochloride, 52) Paraxonase-2, 53) Pocaine, 54) Porfiromycin (POR), 55) Protein, 56) Peptide, 57) Radiosensitizing nucleosides or compounds, 58) Resveratrol, 59) RRx-001, 60) SiRNa, 61) Suppressors of sulfhydral groups, 62) SYM004, 63) Texaphyrins, 64) TH-302, and 65) Tirapazamine.
In one embodiment of the invention, C1FRPC and C2FRPC act synergistically, i.e. that the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising C1FRPC and C2FRPC is larger than the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising only C1FRPC or C2FRPC.
In some cases, the photo-sensitizing or sono-sensitizing or radio-sensitizing strength is the concentration or quantity of radical species, preferentially free radical species, produced or captured by the at least one photo-sensitizer, radio-sensitizer, sono-sensitizer, center of activity or free radical capture or production, or composition or at least one constituent of the composition, or is or is proportional to the strength or intensity of the acoustic wave, ultrasound, radiation, light, electromagnetic radiation, preferentially applied on the photo-sensitizer, radio-sensitizer, sono-sensitizer, center of activity or free radical capture or production or composition or at least one constituent of the composition.
In one embodiment of the invention, C1FRPC and C2FRPC act anti-synergistically, i.e. that the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising C1FRPC and C2FRPC is lower than the photo-sensitizing or sono-sensitizing or radio-sensitizing strength of the nanoparticle or at least one constituent of the composition comprising only C1FRPC or C2FRPC.
In one embodiment of the invention, the composition or at least one constituent of the composition is introduced or administered to or in the body part, preferentially of an animal or human. Preferentially, following its introduction or administration, the nanoparticle or at least one constituent of the composition firstly comprises C1FRPC and C2FRPC during a time t1 and secondly comprises C1FRPC during a time t2, where t2 follows t1.
In some cases, t1 and/or t2 can be longer than 10−10, 10−5, 10−3, 10−1, 0, 1, 5, 10 or 103 second(s) or minute(s).
In some other cases t1 and/or t2 can be shorter than 1010, 105, 103, 10, 0, 1, 5, 10−1 or 10−3 second(s) or minute(s).
In some cases, t2 can be separated from t1 by more than 10−10, 10−5, 10−3, 10−1, 0, 1, 5, 10 or 103 second(s) or minute(s).
In some other cases, t2 can be separated from t1 by less than 1010, 105, 103, 10, 0, 1, 5, 10−1 or 10−3 second(s) or minute(s).
The invention also relates to the composition or at least one constituent of the composition according to the invention, where the composition or at least one constituent of the composition is or comprises: i) a medical device, ii) a drug, iii) a pharmaceutical product or preparation, iv) a medical product or preparation, iv) a biological product or preparation, vi) an adjuvant, vii) an excipient, viii) an active principle, ix) a vaccine or vaccine component, vi) a product, vii) a suspension, viii) a lyophilized suspension or composition or at least one constituent of the composition or preparation, ix) at least one chain of nanoparticle or at least one constituent of the composition, and/or x) at least one center of activity or free radical capture or production.
In one embodiment, the composition or at least one constituent of the composition according to the invention is used for/in: i) the treatment of an infection, ii) the treatment of a viral disease, iii) the treatment of a disease, preferentially a cancerous disease, iv) radiotherapy, v) photodynamic therapy, and/or vi) sonodynamic therapy.
In some cases, the disease can be due to the malfunction of an organ or body part preferentially of an individual.
In some cases, the treatment can be the therapy and/or the diagnosis of a disease or a cosmetic treatment.
In some cases, the treatment can induce the death, destruction, denaturation, or inactivation of at least 1 biological material, such as cell, preferentially pathological cell, RNA, DNA, protein, lipid, or enzyme, where the death of cell can occur through apoptosis or necrosis.
The invention also relates to nanoparticle or at least one constituent of the compositions for use according to the invention, the disease is selected from the group consisting of: a disease associated with a proliferation of cells that is different from the cellular proliferation in a healthy individual, a disease associated with the presence of pathological cells in the body part, a disease associated with the presence of a pathological site in an individual or body part, a disease or disorder or malfunction of the body part, a disease associated with the presence of radio-resistant or acoustic-resistant cells, an infectious disease, an auto-immune disease, a neuropathology, a cancer, a tumor, a disease comprising or due to at least one cancer or tumor cell, a cutaneous condition, an endocrine disease, an eye disease or disorder, an intestinal disease, a communication disorder, a genetic disorder, a neurological disorder, a voice disorder, a vulvovaginal disorder, a liver disorder, a heart disorder, a heating disorder, a mood disorder, anemia, preferentially iron anemia, and a personality disorder.
In some cases, the disease or disorder can be the disease or disorder of or belonging to the individual or body part, or the disease or disorder from which the individual is suffering.
In one embodiment of the invention, the cancer or tumor selected from the group consisting of: the cancer of an organ, cancer of blood, cancer of a system of a living organism, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colon/rectum cancer, endometrial cancer, esophagus cancer, eye cancer, gallbladder cancer, heart cancer, kidney cancer, laryngeal and hypopharyngeal cancer, leukemia, liver cancer, lung cancer, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma cancer, ovarian cancer, pancreatic cancer, pancreatic penile cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, uterine cancer, uterine sarcoma cancer, vaginal cancer, vulvar cancer, waldenstrom macroglobulinemia wilms tumor, castleman disease ewing family of tumor, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, myelodysplastic syndrome pituitary tumor, and a cancerous disease such as gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, malignant mesothelioma, and multiple myeloma.
The invention also relates to a method for the treatment of anemia or anemia disease or to nanoparticle or at least one constituent of the compositions, in particular magnetosomes, for use in the treatment of anemia disease, preferentially iron anemia disease, wherein magnetosomes are administered to the body part of an individual, preferentially to reduce or stop anemia.
The invention also relates to a method for the treatment of an anemia disease, wherein this disease is selected from the group consisting of: Iron or metal deficiency anemia, Vitamin deficiency anemia, Anemia of chronic disease, Aplastic anemia, Anemia associated with bone marrow disease, Hemolytic anemia, Sickle cell anemia, Thalassaemia, Pernicious anaemia, Fanconi anaemia, Sideroblastic Anemia, Congenital Dyserythropoietic Anemia (CDA), Diamond Blackfan Anemia, and Megaloblastic Anemia.
In some cases, anemia is a decrease in the total amount of red blood cells (RBCs) or hemoglobin in the blood, or a lowered ability of the blood to carry oxygen.
The invention also relates to a method of fabrication of the composition or at least one constituent of the composition according to the invention, which comprises at least one of the following steps:
The invention also relates to the method according to the invention, wherein the concentration of the source of zinc, preferentially zinc citrate or zinc sulfate, is comprised between 1 and 100 μM, preferentially 2 and 50 μM, most preferentially 5 and 20 μM.
The invention also relates to the method according to the invention, wherein the nanoparticle or at least one constituent of the compositions preferentially fabricated according to the method comprise a quantity of metal other than iron, preferentially zinc or aluminum, which is incorporated in or comprised in the metallic core is: i) comprised between 0.1 and 100 mg of metal other than iron comprised in the core per gram of iron comprised in the core, ii) preferentially comprised between 0.5 and 10 mg of metal other than iron comprised in the core per gram of iron comprised in the core, iii) most preferentially comprised between 1 and 5 mg of metal other than iron comprised in the core per gram of iron comprised in the core.
In one embodiment of this invention, the nanoparticle or at least one constituent of the composition is or belongs to or is comprised in the group of nanoparticle or at least one constituent of the compositions selected from: a nanosphere, a nanocapsule, a dendrimer, a carbon nanotube, a lipid/solid nanoparticle or at least one constituent of the composition, a lipid or protein or DNA or RNA based nanoparticle or at least one constituent of the composition, a nanoparticle or at least one constituent of the composition with an inner aqueous environment surrounded by a layer, preferentially a stabilizing layer, most preferentially a phospholipid layer, a multilayer nanoparticle or at least one constituent of the composition, a polymeric nanoparticle or at least one constituent of the composition, a quantum dot, a metallic nanoparticle or at least one constituent of the composition, a polymeric micelle or nanoparticle or at least one constituent of the composition, a carbon based nano-structure, a nanobubble, a nanosome, a pharmacyte, a niosome, a nanopore, a microbivore, a liposome, a virus, preferentially recombinant, a herbal nanoparticle or at least one constituent of the composition, an antibody, and a vesicle.
In another embodiment of this invention, the nanoparticle or at least one constituent of the composition is not or does not belong to or is not comprised in at least one nanoparticle or at least one constituent of the composition belonging to the group of: a nanosphere, a nanocapsule, a dendrimer, a carbon nanotube, a lipid/solid nanoparticle or at least one constituent of the composition, a lipid or protein or DNA or RNA based nanoparticle or at least one constituent of the composition, a nanoparticle or at least one constituent of the composition with an inner aqueous environment surrounded by a layer, preferentially a stabilizing layer, most preferentially a phospholipid layer, a multilayer nanoparticle or at least one constituent of the composition, a polymeric nanoparticle or at least one constituent of the composition, a quantum dot, a metallic nanoparticle or at least one constituent of the composition, a polymeric micelle or nanoparticle or at least one constituent of the composition, a carbon based nano-structure, a nanobubble, a nanosome, a pharmacyte, a niosome, a nanopore, a microbivore, a liposome, a virus, preferentially recombinant, a herbal nanoparticle or at least one constituent of the composition, an antibody, and a vesicle.
In some cases, the nanoparticle or at least one constituent of the composition can be in a liquid, gaseous, or solid state, preferentially before, during or after its presence or administration in the body part.
In some other cases, the nanoparticle or at least one constituent of the composition can't be in one or two of the liquid, gaseous, or solid states, preferentially before, during or after its presence or administration in the body part.
In still some other cases, the nanoparticle or at least one constituent of the compositions can be assimilated to or be comprised in a ferrofluid, a chemical or biological ferrofluid, wherein chemical and biological ferrofluids are fluids containing iron, preferentially forming nanoparticle or at least one constituent of the compositions, which are fabricated through a chemical or biological synthesis, respectively.
In still some other cases, the ferrofluid or nanoparticle or at least one constituent of the composition assembly can comprise the nanoparticle or at least one constituent of the compositions and an excipient, a solvent, a matrix, a gel, which preferentially enables the administration of the nanoparticle or at least one constituent of the compositions to the individual or body part.
In still some other cases, the nanoparticle or at least one constituent of the composition can comprise synthetic material and/or biological material and/or inorganic material and/or organic material.
In one embodiment of the invention, (the) nanoparticle or at least one constituent of the composition(s) is/are or designate: i) a suspension of nanoparticle or at least one constituent of the compositions, ii) a composition or at least one constituent of the composition comprising nanoparticle or at least one constituent of the compositions, iii) an assembly of nanoparticle or at least one constituent of the compositions, iv) a nanoparticle or at least one constituent of the composition region, v) the mineral part or core the nanoparticle or at least one constituent of the composition, vi) the organic part of the nanoparticle or at least one constituent of the composition, vii) the inorganic part of the nanoparticle or at least one constituent of the composition, viii) or the coating of the nanoparticle or at least one constituent of the composition.
In one embodiment of the invention, nanoparticle or at least one constituent of the composition(s) or the nanoparticle or at least one constituent of the composition(s) represent(s) or is or are an assembly or suspension or composition or at least one constituent of the composition of more or comprising more than 10−100, 10−50, 10−10, 10−5, 10−1, 1, 10, 102, 103, 105, 1010, 1020 or 1050 nanoparticle or at least one constituent of the composition(s) or mg of nanoparticle or at least one constituent of the composition(s) or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) or mg of nanoparticle or at least one constituent of the composition(s) per cm3 or mg of nanoparticle or at least one constituent of the composition(s) per cm3 of body part or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) per cm3 or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) per cm3 of body part. In some cases, an assembly or suspension or composition or at least one constituent of the composition comprising a large number of nanoparticle or at least one constituent of the compositions can be used to induce or produce a temperature increase, radical or reactive species, or the dissociation of a compound from the nanoparticle or at least one constituent of the compositions.
In another embodiment of the invention, nanoparticle or at least one constituent of the composition(s) or the nanoparticle or at least one constituent of the composition(s) represent(s) or is or are an assembly or suspension or composition or at least one constituent of the composition of less or comprising less than 10100, 1050, 1020, 1010, 105, 102, 10, 1, 5, 2, 1, 10−1, 10−5, 10−10 or 10−50 nanoparticle or at least one constituent of the composition(s) or mg of nanoparticle or at least one constituent of the composition(s) or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) or mg of nanoparticle or at least one constituent of the composition(s) per cm3 or mg of nanoparticle or at least one constituent of the composition(s) per cm3 of body part or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) per cm3 or mg of iron comprised in nanoparticle or at least one constituent of the composition(s) per cm3 of body part. In some cases, an assembly or suspension or composition or at least one constituent of the composition of nanoparticle or at least one constituent of the compositions comprising a low number of nanoparticle or at least one constituent of the composition(s) can be used to prevent toxicity.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition(s) or nanoparticle or at least one constituent of the composition(s) or nanoparticle or at least one constituent of the composition assembly can represent or be the region, also designated as nanoparticle or at least one constituent of the composition region, volume, surface, length, which comprises the nanoparticle or at least one constituent of the compositions or where nanoparticle or at least one constituent of the compositions are located. In some cases, the volume of the region occupied by the nanoparticle or at least one constituent of the compositions in the body part is designated as nanoparticle or at least one constituent of the composition region.
In some cases, the nanoparticle or at least one constituent of the composition region can be the volume occupied by an assembly of nanoparticle or at least one constituent of the compositions in the body part, where the nanoparticle or at least one constituent of the compositions are preferentially separated by less than 109, 106, 103 or 10 nm.
In some cases, the nanoparticle or at least one constituent of the composition assembly is a more general term than nanoparticle or at least one constituent of the composition region, which could designate any type of nanoparticle or at least one constituent of the composition assembly, before, during, or after nanoparticle or at least one constituent of the composition administration to or in the body part.
In some cases, the separating distance between the nanoparticle or at least one constituent of the compositions within the nanoparticle or at least one constituent of the composition assembly or nanoparticle or at least one constituent of the composition region can correspond to the average or maximum distance separating the nanoparticle or at least one constituent of the compositions within this assembly.
In some cases, the distribution in separating distances between nanoparticle or at least one constituent of the compositions can highlight the presence of a minority of nanoparticle or at least one constituent of the compositions, i.e. preferentially less than 50, 10, 1, 10−2 or 10−5% of the total number of nanoparticle or at least one constituent of the compositions in the individual, with either small separating distances, i.e separating distances preferentially lower than 109, 106, 103 or 10 nm, or with large separating distances, i.e. separating distances preferentially larger than 109, 106, 103 or 10 nm. In this case, the presence of this minority of nanoparticle or at least one constituent of the compositions is preferentially not taken into consideration to estimate the average or maximum separating distance between the nanoparticle or at least one constituent of the compositions.
The invention also relates to nanoparticle or at least one constituent of the compositions for use according to the invention, wherein the nanoparticle or at least one constituent of the compositions are crystallized, metallic, or magnetic.
In an embodiment of the invention, the nanoparticle or at least one constituent of the compositions are crystallized. In this case, they preferentially possess more than or at least 1, 2, 10, 102, 103, 106 or 109 crystallographic plane(s) or regular atomic arrangement(s), preferentially observable by electron microscopy.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are metallic. In this case, they contain at least 1, 10, 103, 105 or 109 metallic atom(s) or contain at least 1, 10, 50, 75 or 90% of metallic atoms, where this percentage can be the ratio between the number or mass of metallic atoms in the nanoparticle or at least one constituent of the composition divided by the total number or mass of all atoms in the nanoparticle or at least one constituent of the composition. The nanoparticle or at least one constituent of the compositions, preferentially metal oxide nanoparticle or at least one constituent of the compositions, can also contain at least 1, 10, 103, 105 or 109 oxygen atom(s), or contain at least 1, 10, 50, 75 or 90% of oxygen atoms, where this percentage can be the ratio between the number or mass of oxygen atoms in the nanoparticle or at least one constituent of the compositions divided by the total number or mass of all atoms in the nanoparticle or at least one constituent of the compositions.
In another embodiment of the invention, the metal or metal atom is selected in the list consisting of: Lithium, Beryllium, Sodium, Magnesium, Aluminum, Potassium, Calcium, Scandium, Titanium, Vanadium, Chromium, Manganese, Iron, Cobalt, Nickel, Copper, Zinc, Gallium, Rubidium, Strontium, Yttrium, Zirconium, Niobium, Molybdenum, Technetium, Ruthenium, Rhodium, Palladium, Silver, Cadmium, Indium, Tin, Cesium, Barium, Lanthanum, Cerium, Praseodymium, Neodymium, Promethium, Samarium, Europium, Gadolinium, Terbium, Dysprosium, Holmium, Erbium, Thulium, Ytterbium, Lutetium, Hafnium, Tantalum, Tungsten, Rhenium, Osmium, Iridium, Platinum, Gold, Mercury, Thallium, Lead, Bismuth, Polonium, Francium, Radium, Actinium, Thorium, Protactinium, Uranium, Neptunium, Plutonium, Americium, Curium, Berkelium, Californium, Einsteinium, Fermium, Mendelevium, Nobelium, Lawrencium, Rutherfordium, Dubnium, Seaborgium, Bohrium, Hassium, Meitnerium, Darmstadtium, Roentgenium, Copernicium, Nihonium, Flerovium, Moscovium, and Livermorium or Livermorium atom.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition contains or comprises less than 1, 10, 103, 105 or 109 metallic atom(s) or contains less than 1, 10, 50, 75 or 90% of metallic atoms, where this percentage can be the ratio between the number or mass of metallic atoms in the nanoparticle or at least one constituent of the composition divided by the total number or mass of all atoms in the nanoparticle or at least one constituent of the composition. It can also contain less than 1, 10, 103, 105 or 109 oxygen atom(s), or contain less than 1, 10, 50, 75 or 90% of oxygen atoms, where this percentage can be the ratio between the number or mass of oxygen atoms in the nanoparticle or at least one constituent of the composition divided by the total number or mass of all atoms in the nanoparticle or at least one constituent of the composition.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition is magnetic when it has a magnetic behavior or property, where the magnetic behavior or property is preferentially selected from the group consisting of a diamagnetic, superparamagnetic, paramagnetic, ferromagnetic, and ferrimagnetic behavior or property.
In some cases, the magnetic behavior or property can be observed or exists at a temperature, which is lower than: i) 105, 103, 500, 350, 200, 100, 50, 20, 10, 1, 0.5 or 1 K (Kelvin), ii) the Curie temperature, or iii) the blocking temperature.
In some other cases, the magnetic behavior or property can be observed or exists at a temperature, which is larger than: i) 0.5, 1, 10, 20, 50, 100, 200, 350, 500, 103 or 105 K, ii) the Curie temperature, or iii) the blocking temperature.
In still some other cases, the magnetic behavior or property can be observed or exists at a temperature, which is between 10−20 and 1020 K, or between 0.1 and 1000 K.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions have or are characterized by at least one of the following properties: i) the presence of a core, preferentially magnetic, preferentially mineral, preferentially composed of a metallic oxide such as iron oxide, most preferentially maghemite or magnetite, or an intermediate composition or at least one constituent of the composition between maghemite and magnetite, ii) the presence of a coating that surrounds the core and preferentially prevents nanoparticle or at least one constituent of the composition aggregation, preferentially enabling nanoparticle or at least one constituent of the composition administration in an organism or in the body part or stabilizing the nanoparticle or at least one constituent of the composition core, where coating thickness may preferably lie between 0.1 nm and 10 μm, between 0.1 nm and 1 μm, between 0.1 nm and 100 nm, between 0.1 nm and 10 nm, or between 1 nm and 5 nm, iii) magnetic properties leading to diamagnetic, paramagnetic, superparamagnetic, ferromagnetic, or ferrimagnetic behavior, iv) a coercivity larger than 0.01, 0.1, 1, 10, 100, 103, 104, 105, 109 or 1020 Oe, v) a ratio between remanent and saturating magnetization larger than 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.75, 0.9 or 0.99, vi) a saturating magnetization larger than 0.1, 1, 5, 10 or 50 emu/g, vii) magnetic properties such as coercivity, remanent and saturating magnetization, preferentially measured or observed at a temperature larger than 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K, viii) a crystallinity, i.e. nanoparticle or at least one constituent of the compositions preferentially possessing at least 1, 2, 5, 10 or 100 crystalline plane(s), preferentially observable or measured by electron microscopy, ix) the presence of a single domain, x) a size that is larger than 0.1, 0.5, 1.5, 10, 15, 20, 25, 30, 50, 60, 70, 80, 100, 120, 150 or 200 nm, xi) a size lying between 0.1 nm and 10 μm, between 0.1 nm and 1 μm, between 0.1 nm and 100 nm, between 1 nm and 100 nm, or between 5 nm and 80 nm, xii) a non-pyrogenicity or a pyrogenicity, which preferentially means that nanoparticle or at least one constituent of the compositions possess an endotoxin concentration lower than 1020, 10000, 1000, 100, 50, 10, 5, 2 or 1 EU (endotoxin unit) per mg of nanoparticle or at least one constituent of the composition or per mg of iron comprised in nanoparticle or at least one constituent of the compositions, or which means that nanoparticle or at least one constituent of the compositions do not trigger fever or an increase in whole body temperature larger than 100, 50, 6.6, 5, 3, 2 or 1° C. following their administration to a living organism or body part, xiii) a synthesis by a synthetizing living organism, preferentially by bacteria, xiv) a chemical synthesis, xv) the presence of less than 50, 25, 15, 10, 5, 2 or 1% of organic or carbon material originating from the synthetizing living organism, xv), the presence of more than 99, 95, 80, 70, 60, 50 or 25% of mineral material originating from the synthetizing living organism, or xvi) a specific absorption rate (SAR) that is larger than 1, 10, 1000 or 104 Watt per gram of nanoparticle or at least one constituent of the composition, preferentially measured under the application of an alternating magnetic field of strength preferentially larger than 0.1, 1, 10 or 100 mT, and/or frequency larger than 1, 10, 100 or 1000 KHz, alternatively preferentially measured under the application of the acoustic wave, alternatively under the application of a radiation such as an electromagnetic acoustic, or light radiation.
In another embodiment of the invention, the nanoparticle or at least one constituent of the compositions have or are characterized by at least one of the following properties: i) a coercivity lower than 0.01, 0.1, 1, 10, 100, 103, 104, 105, 109 or 1020 Oe, ii) a ratio between remanent and saturating magnetization lower than 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.75, 0.9 or 0.99, iii) a saturating magnetization lower than 0.1, 1, 5, 10, 50, 200, 1000 or 5000 emu/g, iv) magnetic properties preferentially measured or observed at a temperature lower than 0.1 K, 1 K, 10 K, 20 K, 50 K, 100 K, 200 K, 300 K, 350 K or 3000 K, v) a size that is lower than 0.1, 0.5, 1.5, 10, 15, 20, 25, 30, 50, 60, 70, 80, 100, 120, 150 or 200 nm, vi) the presence of more than 50, 25, 15, 10, 5, 2 or 1% of organic or carbon material originating from the synthetizing living organism, vii) the presence of less than 99, 95, 80, 70, 60, 50 or 25% of mineral material originating from the synthetizing living organism, or xi), a specific absorption rate (SAR) that is lower than 1, 10, 1000 or 104 Watt per gram of nanoparticle or at least one constituent of the composition, preferentially measured under the application of an alternating magnetic field of strength preferentially lower than 0.1, 1, 10, or 100, 200, 500, 103 or 105 mT, and/or of frequency preferentially lower than 1, 10, 100, 103, 105 or 109 KHz, alternatively preferentially measured under the application of the acoustic wave, alternatively under the application of a radiation such as an electromagnetic acoustic, or light radiation.
In some cases, the mineral can be the part of the nanoparticle or at least one constituent of the composition or magnetosome that does not comprise organic material or comprises a low percentage in mass of organic material, preferentially less than 100, 99, 50, 20, 10, 5, 1, 10−1 or 10−2 percent or percent in mass of organic material. The mineral is preferentially the core of the nanoparticle or at least one constituent of the composition.
In some other cases, the mineral can comprise a percentage in mass of organic material larger than 0, 10−50, 10−10, 10−2, 10−1 or 1 percent or percent in in mass of organic material. This can be the case when the purification step unsuccessfully removes the organic material or when organic material is added to the mineral after the purification step.
In some cases, the nanoparticle or at least one constituent of the compositions can be surrounded by a coating. The coating can be made of a synthetic, organic, or inorganic material or of a substance comprising a function selected in the group consisting of carboxylic acids, phosphoric acids, sulfonic acids, esters, amides, ketones, alcohols, phenols, thiols, amines, ether, sulfides, acid anhydrides, acyl halides, amidines, amides, nitriles, hydroperoxides, imines, aldehydes, and peroxides. In some cases, the coating can be made of carboxy-methyl-dextran, citric acid, phosphatidylcholine (DOPC), or oleic acid. In some cases, the coating can enable the dispersion of the nanoparticle or at least one constituent of the compositions in a matrix or solvent such as water, preferentially without aggregation or sedimentation of the nanoparticle or at least one constituent of the compositions. In some cases, the coating can enable internalization of the nanoparticle or at least one constituent of the compositions in cells. In some other cases, the coating can enable: i) to bind two or more nanoparticle or at least one constituent of the composition(s) together preferentially in a chain, ii) to prevent nanoparticle or at least one constituent of the composition aggregation and/or, iii) to obtain uniform nanoparticle or at least one constituent of the composition distribution, and/or iv) to encapsulate or embed at least one center of activity or of free radical/production.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are non-pyrogenic. Non-pyrogenic nanoparticle or at least one constituent of the compositions preferentially: i) comprise less than 10100, 1050, 1020, 108, 105, 103, or 10 EU (endotoxin unit) or EU per cm3 of body part or EU per mg of nanoparticle or at least one constituent of the composition or EU per cm3 of body part per mg of nanoparticle or at least one constituent of the composition, or ii) induce a temperature increase of the individual or body part of less than 105, 103, 102, 50, 10, 5, 4, 3, 2 or 1° C., preferentially above physiological temperature, preferentially before, after or without the application of the acoustic wave or radiation on the nanoparticle or at least one constituent of the composition.
In one embodiment of this invention, the nanoparticle or at least one constituent of the composition or compound is composed of or comprises a chemical element of the families selected from the group consisting of: metals (alkali metal, alkaline earth metal, transition metals), semimetal, non-metal (halogens element, noble gas), chalcogen elements, lanthanide, and actinide.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition or compound is composed of or comprises a chemical element selected from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, caesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, lanthanide, actinide, titanium, zirconium, hafnium, rutherfordium, vanadium, niobium, tantalum, dubnium, chromium, molybdenum, tungsten, seaborgium, manganese, technetium, rhenium, bohrium, iron, ruthenium, osmium, hessium, cobalt, rhodium, iridium, meitherium, nickel, palladium, platinum, darmstadtium, copper, silver, gold, roentgenium, zinc, cadmium, mercury, copernicum, boron, aluminium, gallium, indium, thallium, ununtrium, carbon, silicon, germanium, tin, lead, fleovium, nitrogen, phosphorus, arsenic, antimony, bismuth, ununpentium, oxygen, sulphur, selenium, tellurium, polonium, livermorium, fluorine, chlorine, bromine, iodine, astatine, ununseptium, helium, neon, argon, krypton, xenon, radon, ununoctium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium, actinium, thorium, proctactinium, uranium, neptunium, plutonium, americium, curium, berkelium, californium, einsteinium, fermium, mendelevium, nobelium, and lawrencium.
In some cases, the nanoparticle or at least one constituent of the composition or compound can also be composed of or comprise an alloy, a mixture, or an oxide of this(these) chemical element(s).
In some cases, the nanoparticle or at least one constituent of the composition or compound can be composed of more than 10−50, 10−20, 10−10, 10−5, 10−2, 1, 5, 10, 50, 75, 80, 90, 95 or 99% of one or several of this(these) element(s), where this percentage can represent the mass or number of this(these) chemical elements comprised in the nanoparticle or at least one constituent of the composition or compound divided by the total number or total mass of all chemical elements comprised in the nanoparticle or at least one constituent of the composition or compound or by the total mass of the nanoparticle or at least one constituent of the composition or compound.
In some other cases, the nanoparticle or at least one constituent of the composition or compound can be composed of or comprise less than 10−50, 10−20, 10−10, 10−5, 10−2, 1, 5, 10, 50, 75, 80, 90, 95 or 99% of one or several of this(these) chemical element(s).
In still some other cases, this(these) chemical element(s) is(are) comprised inside the nanoparticle or at least one constituent of the composition or compound, or at the surface of the nanoparticle or at least one constituent of the composition or compound, or in the mineral or core of the nanoparticle or at least one constituent of the composition or compound, or in the coating of the nanoparticle or at least one constituent of the composition or compound.
In one embodiment of this invention, the nanoparticle or at least one constituent of the composition or compound is not composed of or does not comprise at least one chemical element belonging to the family selected from the group consisting of: metals (alkali metal, alkaline earth metal, transition metals), semimetal, non-metal (halogens element, noble gas), chalcogen elements, lanthanide, actinide.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition or compound is devoid of or does not comprise at least one chemical element selected from the group consisting of: hydrogen, lithium, sodium, potassium, rubidium, caesium, francium, beryllium, magnesium, calcium, strontium, barium, radium, scandium, yttrium, lanthanide, actinide, titanium, zirconium, hafnium, rutherfordium, vanadium, niobium, tantalum, dubnium, chromium, molybdenum, tungsten, seaborgium, manganese, technetium, rhenium, bohrium, iron, ruthenium, osmium, hessium, cobalt, rhodium, iridium, meitherium, nickel, palladium, platinum, darmstadtium, copper, silver, gold, roentgenium, zinc, cadmium, mercury, copernicum, boron, aluminium, gallium, indium, thallium, ununtrium, carbon, silicon, germanium, tin, lead, fleovium, nitrogen, phosphorus, arsenic, antimony, bismuth, ununpentium, oxygen, sulphur, selenium, tellurium, polonium, livermorium, fluorine, chlorine, bromine, iodine, astatine, ununseptium, helium, neon, argon, krypton, xenon, radon, ununoctium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium, actinium, thorium, proctactinium, uranium, neptunium, plutonium, americium, curium, berkelium, californium, einsteinium, fermium, mendelevium, nobelium, and lawrencium.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition or compound is not composed of or does not comprise an alloy, a mixture, or an oxide of this(these) chemical element(s).
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition is defined as a particle with a size in one dimension, which is larger than 104, 1, 2, 5, 10, 20, 50, 70, 100, 200 or 500 nm. A nanoparticle or at least one constituent of the composition with a large size can have a larger coercivity and/or a larger remanent magnetization and/or can more strongly or more efficiently absorb the energy or power of the acoustic wave than a nanoparticle or at least one constituent of the composition with a small size. In some cases, the amount of energy or power absorbed by a nanoparticle or at least one constituent of the composition is increased by a factor of more than 1.001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 103, 105 or 107 by increasing the size of the nanoparticle or at least one constituent of the composition by a factor of more than 1.001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 103, 105 or 107.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition is defined as a particle with a size in one dimension, which is lower than 104, 103, 102, 10, 1 or 10−1 nm. Preferentially, a nanoparticle or at least one constituent of the composition with a small size can more easily be administered, for example intravenously, or can enable the avoidance of some toxicity effects, such as embolism.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition size lies between 10−2 and 1020 nm, 10−2 and 104 nm, between 10−1 and 103 nm, or between 1 and 102 nm. This can be the case when the nanoparticle or at least one constituent of the composition or nanoparticle or at least one constituent of the composition assembly possesses a well-defined, preferentially narrow, distribution in sizes.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition size distribution is lower than 1000, 100, 75, 50, 25, 10, 5, 2 or 1 nm. Preferentially, a narrow nanoparticle or at least one constituent of the composition size distribution may be desired to prevent aggregation, or to favor an organization in chains of the nanoparticle or at least one constituent of the compositions.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition size distribution is larger than 1000, 100, 75, 50, 25, 10, 5, 2 or 1 nm. Preferentially, a large nanoparticle or at least one constituent of the composition size distribution may in some cases enable nanoparticle or at least one constituent of the compositions to be eliminated more rapidly.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition has a surface charge, which is larger than −200, −100, −50, −10, −5, 0.1, 1, 2, 5, 10, 50 or 100 mV, preferentially at a pH lower than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14. Preferentially, a nanoparticle or at least one constituent of the composition can have a large surface charge at low pH when it is surrounded by a coating that enables to reach such charge without being destroyed.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition has a surface charge, which is lower than −200, −100, −50, −10, −5, 0.1, 1, 2, 5, 10, 50 or 100 mV, preferentially at a pH larger than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14. Preferentially, a nanoparticle or at least one constituent of the composition can have a low surface charge at high pH when it is surrounded by a coating that enables to reach such charge without being destroyed.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition has a surface charge comprised between +200 and −200 mV, +100 and −100 mV, +50 and −50 mV, +40 et-40 mV, +20 and −20, +10 and −10 mV, or between +5 and −5 mV, preferentially at a pH lower than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the composition has a surface charge comprised between +200 and −200 mV, +100 and −100 mV, +50 and −50 mV, +40 et-40 mV, +20 and −20, +10 and −10 mV, or between +5 and −5 mV, preferentially at a pH larger than 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14.
In another embodiment of the invention, the nanoparticle or at least one constituent of the composition has a weight or a mass, preferentially expressed in unit such as gram (g), kilogram (kg), or milligram (mg). A gram of nanoparticle or at least one constituent of the composition can be a gram of metal such as iron comprised in the nanoparticle or at least one constituent of the composition. The mass or weight of the nanoparticle or at least one constituent of the composition can correspond to the mass or weight of one nanoparticle or at least one constituent of the composition or to the mass or weight of an assembly of nanoparticle or at least one constituent of the compositions.
In an embodiment, the mass of the nanoparticle or at least one constituent of the composition is larger than 10−20, 10−10, 10−5, 10−2, 1, 10, 103, 109 or 1020 gram. In some cases, a large nanoparticle or at least one constituent of the composition mass may be desired to increase the quantity of acoustic wave energy absorbed by the nanoparticle or at least one constituent of the composition.
In an embodiment, the mass of the nanoparticle or at least one constituent of the composition is lower than 10−20, 10−10, 10−5, 10−2, 1, 10, 103, 109 or 1020 gram. In some cases, a low nanoparticle or at least one constituent of the composition mass may be desired to prevent or minimize nanoparticle or at least one constituent of the composition toxicity.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are arranged in chains comprising more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35 or 40 nanoparticle(s) or constituent(s) of the compositions.
In another embodiment of the invention, the nanoparticle(s) or constituent(s) of the compositions are arranged in chains, which have: i) a length smaller than 2·1010, 2·105, 2·103 or 2·102 nm, or ii) a number of nanoparticle or at least one constituent of the compositions in each chain smaller than 2, 5, 10, 102 or 103. In some cases, short chains of nanoparticle or at least one constituent of the compositions may be desired or obtained, for example to favor nanoparticle or at least one constituent of the composition internalization in cells or after partial or total destruction of long chains.
In another embodiment of the invention, the nanoparticle(s) or constituent(s) of the compositions are arranged in chains, which have: i) a length longer than 10−1, 1, 5, 10, 2·102, 2·103 or 2·105 nm or ii) a number of nanoparticle or at least one constituent of the compositions in each chain larger than 2, 5, 10, 102 or 103. In some cases, long chains of nanoparticle or at least one constituent of the compositions may be desired or obtained to increase the quantity of heat or compounds dissociated from the nanoparticle or at least one constituent of the compositions under the application of an acoustic wave or radiation or to prevent nanoparticle or at least one constituent of the composition aggregation or enable uniform nanoparticle or at least one constituent of the composition distribution.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are arranged in chains, which have: i) a length between 10−1 and 1010 nm, or between 1 and 105 nm, or ii) a number of nanoparticle or at least one constituent of the compositions in each chain between 2 and 105, 2 and 103, 2 and 102, or 2 and 50.
In still another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are arranged in chains when they are bound or linked to each other or when the crystallographic directions of two adjacent nanoparticle or constituents of the compositions in the chain are aligned, wherein such alignment is preferentially characterized by an angle between two crystallographic directions belonging to two adjacent nanoparticle or constituents of the compositions in the chains of less than 90, 80, 70, 60, 50, 20, 10, 3, or 2° (degree).
Preferentially when the nanoparticles or at least one constituent of the compositions are biologically synthesized, the nanoparticle or at least one constituent of the compositions can be arranged in chains: i) inside the organism that synthesizes them, also designated as synthetizing living organism, or ii) outside this organism Preferentially, nanoparticle or at least one constituent of the compositions are arranged in chains after or before their extraction or isolation from this organism.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions are not arranged in chains.
In another embodiment of the invention, the nanoparticle or at least one constituent of the compositions are synthesized chemically or are not synthesized by a living organism when less than 1, 2, 5, 10 or 100 step(s) of their production, such as crystallization of iron oxide, stabilization of the iron oxide mineral, organization of the nanoparticle or at least one constituent of the compositions, involves or is due to a living organism In some cases, a chemical synthesis can be defined as a synthesis involving a majority of steps, or more than 1, 2, 5 or 10 steps, or more than 1, 2, 5, 25, 50, 75 or 90% of steps, which involve chemical reactions occurring without the involvement of living organisms, or parts of living organisms such as DNA, RNA, proteins, enzymes, lipids.
In another embodiment of the invention, a chemical synthesis can be used to produce a chemical substance or compound that mimics, copies, or reproduces the compartment, organelle, or other biological material, wherein this chemical synthesis or chemical substance can be used or can result in the production of the nanoparticle or at least one constituent of the compositions. In some cases, the compartment, organelle, or other biological material, can be a lysosome, an endosome, a vesicle, preferentially biological material that has the capacity or the function either to dissolve or transform crystallized iron into free iron or to transform free iron into crystalized iron. In some cases, this transformation is partial and preferentially results in the destruction or formation of partly crystallized assembly of iron atoms or ions, or preferentially results in a mixture of crystallized iron and non-crystallized iron. In some cases, crystallized iron can be defined as an assembly of iron atoms or ions that leads to the presence of crystallographic planes, preferentially observable using a technique such as transmission or scanning electron microscopy as a characterization method, and free iron can preferentially be defined as one of several iron atoms or ions that do not lead to the presence of crystallographic planes, preferentially highlighted by the absence of diffraction patterns, using for example transmission or scanning electron microscopy as a characterization method.
The invention also relates to nanoparticle or at least one constituent of the compositions for use, wherein nanoparticle or at least one constituent of the compositions are or are assimilated to chemical analogues of magnetosomes, such as iron oxide nanoparticle or at least one constituent of the compositions designated as Sigma nanoparticle or at least one constituent of the compositions (ref: 637106-25G), SPION20 (Nanomag®-D-spio 20, Ref: 79-02-201), SPION50 (synomag-D50, Ref: 104-000-501), SPION100 (Nanomag®-D-spio 100, Ref: 79-00-102) or nanoparticle or at least one constituent of the compositions synthesized using a similar method as for these nanoparticle or at least one constituent of the compositions but yielding improved or additional properties such as an arrangement in chains.
In some cases, chemical analogues of magnetosomes can be synthesizes chemically and/or are not synthesized by magnetotactic bacteria.
In some cases, chemical analogues of magnetosomes possess at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 common property(ies) with the magnetosomes, where these common properties are preferentially a ferrimagnetic behavior, preferentially a coercivity larger that 10−50, 10−10, 10−2, 1, 5, 10 or 100 Oe at a temperature preferentially larger than 0, 5, 10, 50, 100, 200, 300, 500 or 1000 K, a large size, preferentially a size larger than 1, 5, 10, 20, 50 or 70 nm, and/or a chain arrangement, preferentially an arrangement of more than 1, 2, 5 or 10 nanoparticle or at least one constituent of the compositions in chain.
In one embodiment of the invention, the nanoparticle or at least one constituent of the compositions or magnetosomes are purified to remove more than 10, 50 or 90 percent or percent in mass of endotoxins and/or other biological material such as proteins or lipids originating from the synthetizing living organism or magnetotactic bacteria. In some other cases, the nanoparticle or at least one constituent of the compositions or magnetosomes are purified to remove less than 100, 99.9, 99, 95 or 90 percent or percent in mass of endotoxins and/or other biological material. This purification step preferentially yields purified nanoparticle or at least one constituent of the compositions or magnetosomes. In some cases, this percentage can be equal to [QBP−QAP]/QBP or QAP/QBP, where QBP and QAP are the quantities of endotoxins, biological material, proteins, or lipids before and after the purification step, respectively.
In some cases, the purification step can consist in using a method or detergent(s) such as NaOH and/or KOH, which is/are preferentially mixed with the synthetizing living organism or magnetotactic bacteria or bacterial debris, preferentially to remove organic material or separate the organic material from the inorganic material comprised in the nanoparticle or at least one constituent of the compositions or magnetosomes and preferentially then be able to harvest the nanoparticle or at least one constituent of the composition or magnetosome mineral, preferentially comprised in the nanoparticle or at least one constituent of the compositions or magnetosomes.
In some cases, the purified nanoparticle or at least one constituent of the compositions or magnetosomes are nanoparticle or at least one constituent of the composition or magnetosome minerals.
In an embodiment of the invention, the nanoparticle or at least one constituent of the compositions according to the invention are drugs, medical devices, cosmetic products, biological products, products used for research purposes, or products used to determine the properties of biological samples.
The invention relates to a method for storing the composition or at least one constituent of the composition according to the invention, which comprises at least one of the following step(s):
The invention relates to the composition or at least one constituent of the composition according to the invention comprising at least one nanoparticle or at least one constituent of the composition, wherein preferentially each nanoparticle or at least one constituent of the composition comprises preferentially an iron oxide mineral core preferentially surrounded by a coating,
The invention also relates to the composition or at least one constituent of the composition according to invention, wherein preferentially an amplifier of radiation is at least one atom that is the center of production of radical species, which is preferentially activated or produces radical species when it is exposed to radiation.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein preferentially an attenuator of radiation is at least one atom that is the center of capture of radical species, which is preferentially activated or captures radical species when it is exposed to radiation.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein CA1 is preferentially a first center of activity or free radical production or capture, C1FRPC, and/or CA2 is preferentially a second center of activity or free radical production or capture, C2FRPC, which is/are preferentially selected in the group consisting of:
The invention relates to the composition or at least one constituent of the composition according to the invention, wherein CA1 and/or CA2 is/are preferentially selected in the group consisting of: 1) Acridine, such as Acridine Orange, acridine yellow, 2) ALA (5-Aminolevulinic acid), 3) Aluminum phthalocyanine tetrasulfonate (AlPcS4), 4) Aminolevulinic acid, delta-Aminolevulinic acid, 5) Antihistamines, 6) Azulene, 7) Bavteriochlorin, 8) TOOKAD or TOOKAD Soluble, 9) WST-11, 10) LUZ11, 11) BC19, 12) BC21, 13) porphyrin such as Benzoporphyrin derivative monoacid ring A (BPD-MA), 14) Chlorin such as Chlorin e6, m-tetrahydroxyphenylchlorin 15) Foscan, 16) Verteporfin, 17) benzoporphyrin derivative mono acid ring A, 18) Monoaspartyl chlorin(e6), 19) talaporfin sodium, 20) HPPH, 21) Transition metal compounds, 22) Chlorine e6 green porphrin, 23) Chlorine e6 porphrin, 24) Coal Tar and Derivatives, 25) Contraceptives, Oral and Estrogens, 26) Curcumin, 27) Cyanine, 28) Cysview, 29) Dyes such as synthetic dyes, 30) Phenothiazinium salts, 31) Rose Bengal, 32) Squaraines, 33) BODIPY dyes, 34) Phenalenones, 35) benzophenoxazinium dyes, 36) Erythrosine, 37) Flavins, 38) Foscan, 39) Fotoscan, 40) Fullerenes such as cationic fullerenes, 41) Furocoumarins, 42) HAL (Hexaminolevulinate), 43) Hemoporfin, 44) 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH), 45) Hypericin, 46) Hypocrellin, 47) ICG (Indocyanine Green), 48) Levulan, 49) MAL-methyl aminolevulinate), 50) Meta-tetra(hydroxyphenyl)chlorin (m-THPC), 51) Metvix, 52) Methylene Blue, 53) Monoterpene, 54) Motexafin lutetium (Lu-Tex), 54) N-aspartyl chlorin e6 (NPe6), 55) Nanoparticle or at least one constituent of the composition or nanomaterial, 56) Natural products or compounds, 57) Non-Steroidal Anti-Inflammatory Drugs, 58) Palladium bacteriopheophorbide (WST09), 59) Phatalocyanin dyes, 60) Phenothiazines, 61) Photochlor, 62) Photofrin, 63) Photosens, 64) Phthalocyanine such as Liposomal ZnPC, 65) Chloroaluminium sulfonated phthalocyanine (CASP), 66) Silicon phthalocyanine (PC4), 67) RLP068, 68) Porfimer sodium, 69) Porfins, 69) Porphyrins, such as 5,10,15,20-Tetrakis(1-methylpyridinium-4-yl) porphyrin tosylate, 70) XF70, 71) Protoporphyrin, 72) ALA-induced protoporphyrin IX, 73) Psoralens, 74) Quantum dots, 75) Quinones, 76) Riboflavin, 77) Rose Bengal, 78) silicon or Silicon phthalocyanine (Pc4), 79) Sulfonamides, 80) Sulfonylureas, 81) Talaporfin or Talaporfin soudium, 82) Temoporfin, 82) Tetrahydropyrroles, 83) Tin ethyl etiopurpurin, 84) Titanium dioxide, 85) Toldudine blue O, 86) Transition metal compounds such as Ruthenium(II), polypyridyl complexes, ruthenium, rhodium, cyclometalated, Rh(II)-Rh(II) bridged dimer compounds, platinum(II), gold(III), 87) Verteporfin, 88) Vulnic based compound such as Aminovulinic, aminovulinic acid, 89) WST11, and 90) Xanthene, 91) ABS-FA, 92) Acrylonitrile Butadiene Styrene, 93) Styrene, 94) Folic acid, 95) AIMP NP, aminoacyl tRNA synthetase complex-interacting multifunctional protein, 96) Au Nanomaterial, 97) gold, 98) Au—MnO nanomaterial, 99) manganese oxide, 100) Antineoplastic drugs, 101) NSAIDs, 102) nonsteroidal anti-inflammatory drug, 103) Artemether, 104) 5-ALA (5-aminolevulinic acid), 105) Acridine, Acridine Orange, 106) Au-doped TiO2, 107) Carbon based nanomaterial, 108) carbon nanotube, 109) Chlorine, 110) Ce6, 111) PTX, Paclitaxel, 112) chemotherapeutic drug or compound, 113) infrared dye or IR783, 114) Curcumin, 115) Cyanine or Cu-Cyanine, 116) DHMS, 117) dimethylsulfure, 118) Docetaxel, 119) chemotherapeutic drug or compound, 119) DOX/Mn-TPPS @RBCS, 120) doxorubicin, 121) manganese, 122) blood cell, 123) red blood cell, cell, 124) polymer, 125) elastomer, 126) Erythosin or Erythosin B, 127) FA or FA-OI or FA-OI NP or folic acid, 128) F3-PLGA@MB/Gd NPs, 129) poly(lactic-co-glycolic acid), 130) gadolinium, 131) Fe—TiO2 or titanium oxide, 132) Fe-VS2, 133) iron, 134) vanadium disulfide, 135) FMSNs-DOX, 136) silica, 137) HCQ, 138) hydrochloroquine, 139) HP, 140) hematoporphyrin, 141) HMME, 142) hematoporphyrin monomethyl ether, 143) HSYA or Hydroxysafflor yellow A, 144) Hypocrellin, Hypocrellin B, 145) IR780, 146) Levofloxacin, 147) LIP3 or Lithium phosphide, 148) Lithium, 149) Liposome or Liposomal nanomaterial, 150) Lomefoxacin, 151) MG@P NPs, 152) MnP or Manganese peroxidase, 153) MnTTP-HSAs, 154) HSA-wrapped metal-porphyrin complex, 155) albumin, 156) MnWOx, 157) MnWOx-PEG, 158), PEG, 159) metallic or bimetallic or multi-metallic compound preferentially oxide, 160) Mn (III)-HFs, 161) managense, 162) hemoporfin, 163) nano-compound or nanoroad or nanoflower or nanowire or quantum dot, 164) Noble or halogen or Hydrogen or alkali metals or Alkaline earth metals or Triels or Tetrels or Pnicto-gen or Chal-co-gens or metal or gas or liquid or solid preferentially nanomaterial, 165) oxygen indyocyanine preferentially nanoparticle or at least one constituent of the composition, 166) Phthalocyanines, 167) PIO or Pioglitazone, 168) Polymeric nanomaterial, 169) Porphyrin, 170) Pt-doped TiO2, 171) R837, 172) Rose Bengal, 173) Sparfloxacin, 174) TAPP or 5,10,15,20-tetrakis (4-aminophenyl) porphyrin, 175) TiO2 or titanium dioxide nanomaterial, 176) TCPP, isomer, or Tris(1-chloro-2-propyl) phosphate 177) TPI or Thermoplastic Polyimide or thermoplastic polymer, 178) TPZ or Tirapazamine, 179) Transition metal oxide, 180) nanoparticle or at least one constituent of the composition or Janus nanoparticle or at least one constituent of the composition, and 181) Xanthones, 182) AQ4N, 183) Apaziquone (E09), 184) Bromodeoxyuridine, 185) Carbogen, 186) Cetuximab, 187) Chemotherapeutic drug or compound, 188) Chlorpromazine, 189) C-reactive peptide, 190) Curcumin, 191) Diamide, 192) Diethylmaeate, 193) Dihydroartemisinin, 194) Docetaxel, 195) ECI301, 196) Etanidazole, 197) Fludarabine, 198) 5-Fluorouracil, 199) Fluorodeoxyuridine, 200) Gadolynium, 201) Gemcitabine, 202) HER-3 ADC, 203) HSP, 204) Hydrogen peroxide, 205) Hydroxyurea, 206) Hyperbaric oxygen, 207) Hyperthermia, 208) Hypoxic cell cytotoxic agent, 209) Irinotecan, 210) lanthanide-doped radiosensitizer-based metal-phenolic network, 211) Lidocaine, 212) Lododeoxyuridine, 213) Metronidazole, 214) misonidazole, 215) etanidazole, 216) nimorazole, 217) N-Ethylmalemide, 218) malmeide, 219) ethylmalmeide, 220) Nanomaterial such as those consisting of or composed of at least partly or fully gold, silver, bismuth, gadolinium, polysiloxane matrix and gadolinium chelates, hafnium, Tantalum, Zinc, Gadolinium, Germanium, Chromium, Praseodymium, Silicon, iron, platinum, cobalt, manganese, magnesium, iron, Titanium, carbon nanotube, quantum dot, nanoroad, Triflate, or metal oxide, 221) Nelfinavir, 222) Nicotinamide, 223) Nimotuzumab, 224) RNA, or miRNA, or miR-201, or miR-205, or miR-144-5p, or miR-146a-5p, or miR-150, or miR-99a, or miR-139-5p, or miR-320a, 225) Membrane active agent, 226) Mitomycin-C or Mitomycin, 227) Motexafin, 228) NBTXR3, 229) Oligonucleotide, 230) Paclitaxel, 231) Papaverine or Papaverine hydrochloride, 232) Paraxonase-2, 233) Pocaine, 234) Porfiromycin (POR), 235) Protein, 236) Peptide, 237) Radiosensitizing nucleosides or compounds, 238) Resveratrol, 239) RRx-001, 240) SiRNa, 241) Suppressors of sulfhydral groups, 242) SYM004, 243) Texaphyrins, 244) TH-302, and 245) Tirapazamine.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein at least two nanoparticle or at least one constituent of the compositions in the composition preferentially arrange in a chain or remained arranged in a chain, preferentially at or below 0° C.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the nanoparticle or at least one constituent of the composition core is preferentially predominantly or in majority metallic, and preferentially the coating of the nanoparticle or at least one constituent of the composition is predominantly or in majority organic or non-metallic.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein preferentially the core is synthesized by a living organism or nanoparticle or at least one constituent of the composition producing cells, and/or preferentially the coating is not synthesized by a living organism or nanoparticle or at least one constituent of the composition producing cells,
wherein the nanoparticle or at least one constituent of the composition producing cell is preferentially a magnetotactic bacterium.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein preferentially C1FRPC and/or C2FRPC is/are is/are:
The invention also relates to a method of fabrication of the composition or at least one constituent of the composition according to the invention, which comprises at least one of the following steps:
Step 1 of preferentially amplifying magnetotactic bacteria, preferentially in at least one medium, comprising:
Step 2 of preferentially extracting magnetosomes from magnetotactic bacteria and/or of preferentially purifying the extracted magnetosomes preferentially by heating them preferentially to yield magnetosome minerals preferentially comprising a percentage in mass of organic material coming from magnetotactic bacteria that is preferentially lower than 50 or 10 or 1%,
Step 3 of preferentially coating the magnetosome minerals preferentially with a coating material comprising the compound CA2 or C2FRPC by mixing the magnetosome minerals with the coating material, where the mixing is preferentially realized in at least one of the following conditions:
Step 4 of preferentially adding at least one cryoprotectant or protectant compound or protectant compound to the coated magnetosome minerals obtained at the end of step 3,
Step 5 of preferentially lyophilizing or dehydrating or drying or desiccating the composition or at least one constituent of the composition obtained at the end of step 4,
Step 6 of preferentially re-suspending the lyophilized composition or at least one constituent of the composition obtained of step 5, preferentially in water.
The invention also relates to a method for storing the composition or at least one constituent of the composition according to invention, which preferentially comprises at least one of the following step(s):
In some cases, the at least one constituent of the composition is selected in the group consisting of: i) the nanoparticle or at least one constituent of the composition, ii) the nanoparticle or at least one constituent of the composition coating, iii) the nanoparticle or at least one constituent of the composition core, iv) the cryoprotectant or protectant compound, v) the other compound, vi) at least one link or bond or force or interaction of or between at least one or two constituent(s) of the composition, vii) a solute, viii) a solvent, ix) an excipient, x) an active part or principle, xi) an inert part, xii) an organic or carbon or carbonaceous part, xiii) an inorganic or metallic part, xiv) a medical device or drug, xv) a pharmaceutical compound, xvi) an immunological compound, xvii) a metabolic compound, xviii) a chemotherapeutic compound, xix) a surgical compound, xx) a radio-sensitizer, xxi) a contrast agent, and xxii) a sonosensitizer of the composition or at least one constituent of the composition.
In some cases, the constituent is comprised in the organic or inorganic part of the composition or at least one constituent of the composition.
In some cases, the constituent is comprised in the inert or active part of the composition or at least one constituent of the composition.
The invention also relates to a composition or at least one constituent of the composition comprising at least one nanoparticle or at least one constituent of the composition or at least one chain of at least two nanoparticle or at least one constituent of the compositions,
The invention also relates to the composition or at least one constituent of the composition according to the invention comprising at least one nanoparticle or at least one constituent of the composition or at least one constituent of the composition or at least one constituent of the composition or at least one chain of at least two nanoparticle or at least one constituent of the compositions, wherein preferentially each nanoparticle or at least one constituent of the composition in the chain comprises an iron oxide mineral core surrounded by a coating,
In one embodiment, the protectant compound is selected in the group consisting of: i) a cryoprotectant or protectant compound, ii) a thermo-protectant, iii) an oxydo-protectant, iv) a chain-protectant, v) an activity-protectant and vi) a protectant of the composition or at least one constituent of the composition or size or cohesion or at least one property or magnetic property of at least one nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition or constituent of the composition or at least one constituent of the composition.
In some cases, an activity-protectant compound is a compound that protects the activity, preferentially a therapeutic, immunological, pharmacological, chemotherapeutical, metabolic, thermal, and/or vaccine-type activity of the nanoparticle or at least one constituent of the composition or of at least one constituent of the composition, i.e. preferentially without the activity-protectant compound the activity of the nanoparticle or at least one constituent of the composition is diminished or decreased or lost partly or fully preferentially over time or under thermal variation or under the application of a radiation on the composition or at least one constituent of the composition.
In some cases, the composition or at least one constituent of the composition can comprise an active part and an inactive part or different parts with different types of activity, for example one part with a medical or thermal activity such as the part comprising the nanoparticle or at least one constituent of the composition or nanoparticle or at least one constituent of the composition core or nanoparticle or at least one constituent of the composition coating and another part with an activity consisting in preserving the at least one property of the nanoparticle or at least one constituent of the composition such as the part comprising the cryoprotectant or protectant compound.
In some cases, the composition or at least one constituent of the composition is of high purity, preferentially in terms of metallic composition or of composition in iron.
In some cases, a high purity composition or at least one constituent of the composition designates a composition or at least one constituent of the composition that comprises more than 50, 90, 96, 99 or 99% of iron preferentially in terms of metallic composition or at least one constituent of the composition.
In some cases, a cryoprotectant or protectant compound can be a compound that protects or maintains or avoids a change of at least one property of the composition or at least one constituent of the composition or nanoparticle, preferentially when the composition or nanoparticle or at least one constituent of the composition is cooled down, preferentially by at least or below 100, 10, 0, −1, −20, −50, −100, or −273° C.
In some cases, a thermo-protectant can be a compound that protects or maintains or avoids a change of at least one property of the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition, preferentially when the composition or at least one constituent of the composition or nanoparticle is subject to a temperature gradient, preferentially by at least 0, 1, 2, 5, 10 or 100° C.
In some cases, an oxydo-protectant can be a compound that protects or maintains or avoids a change of at least one property of the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition such as the oxidation state of the nanoparticle or at least one constituent of the composition or composition, preferentially when the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition is subject to oxidation or reduction.
In some cases, a chain-protectant can be a compound that protects or maintains or avoids a change or destruction or diminution or increase or variation in size of the chain of nanoparticle or at least one constituent of the composition, preferentially when the composition or at least one constituent of the composition or nanoparticle is subject to oxidation or reduction or subject to a temperature gradient or is cooled down or is exposed to a radiation.
In some cases, a size-protectant can be a compound that protects or maintains or avoids a change or destruction or diminution or increase or variation in size of the nanoparticle or at least one constituent of the composition, preferentially by more than 0, 1, 10, 100, 103 or 105 nm, preferentially when the composition or at least one constituent of the composition or nanoparticle is subject to oxidation or reduction or subject to a temperature gradient or is cooled down or is exposed to a radiation.
In some cases, a composition or at least one constituent of the composition-protectant can be a compound that protects or maintains or avoids a change or destruction or diminution or increase or variation in composition of the nanoparticle or at least one constituent of the composition, preferentially by more than 1, 10, 99, 100, 103, 105 or 1010 atoms or percent of atoms, preferentially when the composition or at least one constituent of the composition or nanoparticle is subject to oxidation or reduction or subject to a temperature gradient or is cooled down or is exposed to a radiation.
In some cases, a cohesion-protectant can be a compound that protects or maintains or avoids a change or destruction or diminution or increase or variation in cohesion of the nanoparticle or at least one constituent of the composition or composition, preferentially when the composition or at least one constituent of the composition or nanoparticle is subject to oxidation or reduction or subject to a temperature gradient or is cooled down or is exposed to a radiation.
In some cases, the cohesion of the nanoparticle or at least one constituent of the composition can be at least one interaction or at least one bond or interaction or force or link between at least two constituents of the composition, which preferentially maintain the cohesion or assembly or at least one property or the content of the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition.
In some cases, a magnetic property-protectant can be a compound that protects or maintains or avoids a change or destruction or diminution or increase or variation in magnetic property or coercivity or magnetization or saturating magnetization of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition, preferentially by more than 1, 10, 99, 100, 103, 105 or 1010 Oe or mT or percent of this property, preferentially when the composition or at least one constituent of the composition or nanoparticle or at least one constituent of the composition is subject to oxidation or reduction or subject to a temperature gradient or is cooled down or is exposed to a radiation.
In some cases, at least one magnetic property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition can be a diamagnetic, paramagnetic, superparamagnetic, ferromagnetic or ferrimagnetic property.
In some cases, the at least one property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition exists or is measured at a temperature lower than 1010, 105, 103, 100, 50, 10, 5, 2, 1, 0.1 or 0 K (Kelvin) or ° C. (Celsius degree).
In some cases, the at least one property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition exists or is measured at a temperature larger than 0, 0.1, 1, 5, 10, 50, 100, 103, 105 or 1010 K (Kelvin) or ° C. (Celsius degree).
In still some other cases, the nanoparticle or at least one constituent of the composition can result from the assembly of atoms, entities, compounds that don't have a nanometer size, preferentially before assembly or taken individually, or are preferentially smaller than 100, 10, 1 or 0.1 nm, and preferentially display a nanometer size following their assembly, or are preferentially larger than 0.1, 1, 10 or 100 nm.
In still some other cases, the nanoparticle or at least one constituent of the composition can result from the dis-assembly of atoms, entities, compounds that don't have a nanometer size, preferentially before dis-assembly, or are preferentially larger than 0.1, 1, 10 or 100 nm 1 μm, and preferentially display a nanometer size following their dis-assembly, or are preferentially smaller 1 μm, 100, 10, 1 or 0.1 nm.
In still some other cases, assembly and/or dis-assembly occur(s) inside the body part or outside the body part.
In still some other cases, the composition or at least one constituent of the composition comprises at least one chemical element or a majority of such element or a percentage of mas of such element larger than or equal to 0, 1, 5, 10, 50, 90, or 99%.
In still some other cases, the composition or at least one constituent of the composition comprises at least one chemical element or a minority of such element or a percentage of mas of such element smaller than or equal to 100, 99, 75, 50, 10, 5, 2, 1 or 0%.
In still some other cases, the chemical element is selected in the group consisting of: Actinium, Aluminum, Americium, Antimony, Argon, Arsenic, Astatine, Barium, Berkelium, Beryllium, Bismuth, Bohrium, Boron, Bromine, Cadmium, Calcium, Californium, Carbon, Cerium, Cesium, Chlorine, Chromium, Cobalt, Copernicium, Copper, Curium, Darmstadtium, Dubnium, Dysprosium, Einsteinium, Erbium, Europium, Fermium, Flerovium, Fluorine, Francium, Gadolinium, Gallium, Germanium, Gold, Hafnium, Hassium, Helium, Holmium, Hydrogen, Indium Iodine, Iridium, Iron, Krypton
Lanthanum, Lawrencium, Lead, Lithium, Livermorium, Lutetium, Magnesium,
Manganese, Meitnerium, Mendelevium, Mercury, Molybdenum, Moscovium, Neodymium, Neon, Neptunium, Nickel, Nihonium, Niobium, Nitrogen, Nobelium, Oganesson, Osmium, Oxygen, Palladium, Phosphorus, Platinum, Plutonium, Polonium, Potassium, Praseodymium, Promethium, Protactinium, Radium, Radon, Rhenium, Rhodium, Roentgenium, Rubidium, Ruthenium, Rutherfordium, Samarium, Scandium, Seaborgium, Selenium, Silicon, Silver, Sodium, Strontium, Sulfur, Tantalum, Technetium, Tellurium, Tennessine, Terbium, Thallium, Thorium, Thulium, Tin, Titanium, Tungsten, Uranium, Vanadium, Xenon, Ytterbium, Yttrium, Zinc, and Zirconium.
In one embodiment of the invention, the composition or at least one constituent of the composition comprises the at least one nanoparticle or at least one constituent of the composition with the cryoprotectant or protectant compound or protectant compound and optionally a liquid such as water or a gas or a solid or at least one substance that is different from the nanoparticle or at least one constituent of the composition and/or cryoprotectant or protectant compound. Such substance preferentially disperses or suspends the at least one nanoparticle or at least one constituent of the composition preferentially with the cryoprotectant or protectant compound. Such substance and/or cryoprotectant and/or protectant compound preferentially embeds and/or surrounds and/or act(s) as a matrix surrounding the at least one nanoparticle or at least one constituent of the composition and/or cryoprotectant or protectant compound. Such substance preferentially has a different function than the cryoprotectant or protectant compound. For example, the substance could enable the dispersion or re(dispersion), preferentially homogenously, of the at least one nanoparticle or at least one constituent of the composition, preferentially before administration of the composition or at least one constituent of the composition to the body part, whereas the cryoprotectant or protectant compound could protect or maintain at least one property of the nanoparticle or at least one constituent of the composition, preferentially during storage of the composition or at least one constituent of the composition preferentially in a lyophilized or dehydrated or powder form.
In one embodiment of the invention, the at least one property of the composition or of at least one constituent of the composition is selected in the group consisting of: i) the chain arrangement or geometric figure or assembly of the at least two nanoparticles or constituents of the composition, ii) the composition of at least one constituent of the composition, preferentially maintained by maintaining at least 1, 5, 10, 50, 75, 100, 103 or 105 atom(s) or metal(s) or % of atom(s) or metal(s) or % in mass of atom(s) or metal(s) preferentially in the composition or at least one constituent of the composition, iii) the size of the nanoparticle or at least one constituent of the composition, preferentially by preventing a change in nanoparticle or at least one constituent of the composition size or in size of the at least one constituent of the composition or at least one constituent of the composition, preferentially by more than 1, 5, 10, 100, 103 or 105 nm, iii) the oxidation state of the nanoparticle or at least one constituent of the composition or of the at least one constituent of the composition or at least one constituent of the composition, iv) the at least one magnetic property, preferentially the ferrimagnetic property, of the nanoparticle or at least one constituent of the composition or of the at least one constituent of the composition or at least one constituent of the composition, v) the at least one center of activity or of free radical capture or production of the nanoparticle or at least one constituent of the composition or of the at least one constituent of the composition or at least one constituent of the composition, and vi) the isotonicity or osmolality of the composition or at least one constituent of the composition or of the at least one constituent of the composition or at least one constituent of the composition, wherein preferentially the at least one property exists or is measured preferentially in some cases at or below the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, −5, −10, −50, −77, −100, −200 or −273° C., in some other cases at or above the temperature of 105, 103, 102, 50, 20, 10, 5, 2, 1, 0, −5, −10, −50, −77, −100, −200 or −273° C.
In another embodiment of the invention, the at least one property of the composition or at least one constituent of the composition is maintained, or kept or protected, preferentially by the cryoprotectant or protectant compound or protectant compound, when:
In some cases, the percentage of variation of the at least one other property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition can be equal to (P2−P1)/P1, preferentially measured in absolute terms or values, where P1 and P2 are two different values of at least one other property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition, preferentially measured when the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition is at two different temperature or in two different conditions.
In some cases, the nanoparticle or at least one constituent of the composition is at least one of the following conditions: i) liquid, ii) solid, and iii) gaseous.
In some cases, the property of at least one nanoparticle or at least one constituent of the composition or chain can be: i) the size, diameter, surface or volume, of the nanoparticle or at least one constituent of the composition, ii) at least one magnetic property of the at least one nanoparticle or at least one constituent of the composition, iii) the composition or at least one constituent of the composition preferentially metallic and/or organic or carbonaceous of the at least one nanoparticle or at least one constituent of the composition, iv) the cohesion of the nanoparticle or at least one constituent of the composition, v) the percentage in mass of at least one metal in the at least one nanoparticle or at least one constituent of the composition or chain, vi) the chain arrangement of the at least one nanoparticle or at least one constituent of the composition, vii) the number of nanoparticle or at least one constituent of the compositions preferentially in the chain or composition, viii) the surface charge of the at least one nanoparticle or at least one constituent of the composition or chain, ix) the percentage of water or humidity or solid or liquid or gaseous material in the at least one nanoparticle or at least one constituent of the composition or chain, x) the oxidation or redox state of the at least one nanoparticle or at least one constituent of the composition, xi) the coercivity or remanent magnetization or magnetization or saturating magnetization of the at least one nanoparticle or at least one constituent of the composition or chain, xiii) the core diameter or size or volume or surface area or ratio surface/volume of the at least one nanoparticle or at least one constituent of the composition, xiii) the coating thickness of the at least one nanoparticle or at least one constituent of the composition, and/or xiv) the isotonicity or osmolality of the at least one nanoparticle or at least one constituent of the composition or chain.
In some cases, the at least one nanoparticle or at least one constituent of the composition and preferentially the cryoprotectant or protectant compound can be surrounded by or embedded in or mixed with water or liquid or solid or gas or matrix, or surfactant or solvent, which preferentially enables or favors or triggers the dispersion or suspension, preferentially within a homogenous manner, of the at least one nanoparticle or at least one constituent of the composition preferentially of or in the composition or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the composition or at least one constituent of the composition comprises an organic part and/or an inorganic part, wherein the inorganic part preferentially comprises the core of the nanoparticle or at least one constituent of the composition, wherein the organic part preferentially comprises the coating of the nanoparticle or at least one constituent of the composition and/or the cryoprotectant or protectant compound, and wherein the percentage in mass of the inorganic part is preferentially larger, preferentially a factor ξ, than the percentage in mass of the organic part.
In some cases, the organic part comprises more than 0, 10−10, 1, 5, 10, 35, 50, 75, 80, 90 or 100% in mass or volume of carbon or carbonaceous material.
In some other cases, the inorganic part comprises less than 100, 99, 80, 75, 50, 25, 10, 5, 2, 1 or 0% in mass or volume of carbon or carbonaceous material.
In still some other cases, the organic part comprises more carbon or carbonaceous material than the inorganic part.
In some cases, the factor ξ is equal to I2/O1, where I2 is the percentage in mass of the inorganic part in the composition or at least one constituent of the composition and O1 is the percentage in mass of the organic part in the composition or at least one constituent of the composition, preferentially measured in the absence of or with minimal water or liquid or gas.
In some cases, ξ is larger than or equal to 1, 2, 5, 10, 100, 103 or 105.
In some other cases, ξ is smaller than or equal to 105, 103, 100, 10, 5, 2 or 1.
The invention also relates to the composition or at least one constituent of the composition according to the invention, comprising i) and/or ii):
In some cases, the percentages in mass of the nanoparticle or at least one constituent of the composition, nanoparticle or at least one constituent of the composition core, nanoparticle or at least one constituent of the composition coating and/or cryoprotectant or protectant compound in the composition or at least one constituent of the composition is/are larger than or equal to 0, 10−10, 1, 5, 10, 25, 30, 50, 75, 80, 90, 99.99 or 100%.
In some other cases, the percentages in mass of the nanoparticle or at least one constituent of the composition, nanoparticle or at least one constituent of the composition core, nanoparticle or at least one constituent of the composition coating and/or cryoprotectant or protectant compound in the composition or at least one constituent of the composition is/are smaller than or equal to 100, 99.99, 90, 80, 70, 75, 50, 25, 10, 1, 0.1 or 0%.
In some cases, a1=PM1/PM2, where PM1 is the percentage in mass of the nanoparticle or at least one constituent of the composition core and PM2 is the percentages in mass of the nanoparticle or at least one constituent of the composition coating and/or cryoprotectant or protectant compound.
In some other cases, a2=PM3/PM4, where PM3 is the percentage in mass of carbon or carbonaceous material of the nanoparticle or at least one constituent of the composition core and PM4 is the percentage in mass of carbon or carbonaceous material of the nanoparticle or at least one constituent of the composition coating and/or cryoprotectant or protectant compound.
In still some other cases, a3=PM5/PM6, where PM5 is the percentage in iron or metal mass of the nanoparticle or at least one constituent of the composition core and PM6 is the percentage in iron or metal mass of the nanoparticle or at least one constituent of the composition coating and/or cryoprotectant or protectant compound.
In some cases, a1, a2, and/or a3 is/are larger than or equal to 0, 10−10, 1, 5, 10, 25, 50, 100, 103 or 105.
In some other cases, a1, a2, and/or a3 is/are smaller than or equal to 1040, 1010, 100, 50, 25, 10, 5, 2, 1 or 0.
In one embodiment of the invention, the strength of the bonds or interactions between the coating and core of the nanoparticle or at least one constituent of the composition is strong when the core and coating of the nanoparticle or at least one constituent of the composition can be attracted or moved preferentially together by a magnet preferentially of strength stronger than 10−6, 10−3, 10−1 or 1 T. In some cases, such bonds or interactions can be designated as bonds or interactions that ensure the cohesion of the nanoparticle or at least one constituent of the composition preferentially by being resistant to or maintained in the presence of a force preferentially a magnetic force preferentially applied by a magnet preferentially of strength larger than 10−6, 10−3, 10−1 or 1 T.
In another embodiment of the invention, the strength of the bond or interactions between the cryoprotectant or protectant compound and the coating and/or core of the nanoparticle or at least one constituent of the composition is weak when the cryoprotectant or protectant compound can't be attracted or moved preferentially together by a magnet preferentially of strength stronger than 10−6, 10−3, 10−1 or 1 T while the core and coating of the nanoparticle or at least one constituent of the composition can be attracted or moved preferentially together by a magnet preferentially of strength stronger than 106, 103, 10−1 or 1 T.
In another embodiment of the invention, the strength of the bonds or interactions between the coating and core of the nanoparticle or at least one constituent of the composition is stronger than the strength of the bond or interaction between the cryoprotectant or protectant compound and the coating and/or core of the nanoparticle or at least one constituent of the composition, when the cryoprotectant or protectant compound can be less or less strongly attracted or moved by a magnet preferentially of strength stronger than 10−6, 10−3, 10−1 or 1 T than the nanoparticle or at least one constituent of the composition.
The invention also relates to the composition or at least one constituent of the composition according to the invention, further comprising a compound, preferentially designated as the other compound, which has at least one property selected in the group consisting of:
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the composition or at least one constituent of the composition is in the form of: i) a powder, ii) a liquid, iii) or a liquid suspension, iv) a solid, and/or v) one or a mixture of liquid, solid and/or gaseous state(s).
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the at least one chain of at least two nanoparticle or constituents of the compositions has at least one property selected in the group consisting of:
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the nanoparticle or at least one constituent of the composition comprises at least one center of activity, preferentially of medical activity, preferentially selected in the group consisting of:
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the nanoparticle or at least one constituent of the composition, preferentially the nanoparticle or at least one constituent of the composition core, is synthesized by a living organism or nanoparticle or at least one constituent of the composition producing cell, preferentially a magnetotactic bacterium.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the center of activity is selected in the group consisting of:
The invention also relates to the composition or at least one constituent of the composition according to the invention combined with at least one nanoparticle or at least one constituent of the composition-producing cell, preferentially a magnetotactic bacterium,
In some cases, the first and second cryoprotectant or protectant compounds can be the same or different compounds.
In some other cases, the combination of the composition or at least one constituent of the composition and nanoparticle or at least one constituent of the composition-producing cell is or belongs to a system or combined system or combined product.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the composition or at least one constituent of the composition preferentially comprises an inert part and/or an active part, wherein the inner part preferentially does not comprise at least one center of activity or the active part preferentially comprises at least one center of activity or the active part preferentially comprises more or a larger number of centers of activity than the inner part, wherein the inner part preferentially ensures the cohesion of the composition or at least one constituent of the composition or preferentially comprises links or bonds or forces or atoms or ions or nanoparticle or at least one constituent of the compositions that maintain the at least one or two constituent(s) of the composition assembled together preferentially within one volume, wherein the center of activity is preferentially comprised in the nanoparticle or at least one constituent of the composition, wherein the center of activity preferentially increases or decreases or amplifies or attenuates the production of heat or cold by the nanoparticle or at least one constituent of the composition, the medical activity of the nanoparticle or at least one constituent of the composition, and/or the radiation or strength or power or wavelength or intensity or frequency of the radiation applied on the nanoparticle or at least one constituent of the composition,
In one embodiment, at least one binding material binds at least two nanoparticles or constituents of the compositions preferentially in chain or geometric figure or assembly.
In some cases, the binding material can be the same material as the coating and/or have at least one property in common with the coating or at least one constituent of the composition or at least one constituent of the composition.
In some cases, the binding material can be a different material from the coating, and/or comprise water or a gel or tissue or cellular or cellular component such as cytoplasm and/or have at least one property different from the coating or at least one constituent of the composition or at least one constituent of the composition.
In one embodiment of the invention, the bonds between: i) the nanoparticle or at least one constituent of the composition different from the cryo-protectant or protectant compound, nanoparticle core, and/or nanoparticle coating and ii) the cryoprotectant or protectant compounds are weak bonds, preferentially Van der Walls bonds, preferentially non-covalent or non-metallic bonds.
In one embodiment of the invention, the bonds between: i) the nanoparticle or at least one constituent of the composition, nanoparticle core, and/or nanoparticle coating and ii) the cryoprotectant or protectant compounds are weaker or weaker in strength than the bonds between the coating and the core of the nanoparticle or at least one constituent of the composition.
In one embodiment of the invention, there exists partly or fully a complexation between the core and the coating of the nanoparticle or at least one constituent of the composition, preferentially within part or the whole lifespan of the composition or at least one constituent of the composition, preferentially during lyophilization or storage of the composition or at least one constituent of the composition, preferentially when the composition or at least one constituent of the composition is in liquid, gas, or solid form, or in one of these sates in a dominant manner.
In another embodiment of the invention, there does not exist partly or fully a complexation between the coating or core of the nanoparticle or at least one constituent of the composition and the cryoprotectant or protectant compound, preferentially within part or the whole lifespan of the composition or at least one constituent of the composition, preferentially during lyophilization or storage of the composition or at least one constituent of the composition, preferentially when the composition or at least one constituent of the composition is in liquid, gas, or solid form, or in one of these sates in a dominant manner.
In one embodiment of the invention, the cryoscopic constant of the composition or at least one constituent of the composition or the Ebullioscopic constant of the composition or at least one constituent of the composition or the depression of freezing point of the solvent or of the composition or at least one constituent of the composition is smaller than 10−10, 105, 103, 100, 50, 20, 10, 5, 2, 1, 0 kg×K/mol.
In one embodiment of the invention, the cryoscopic constant of the composition or at least one constituent of the composition, or the Ebullioscopic constant of the composition or at least one constituent of the composition or the depression of freezing point of the solvent or of the composition or at least one constituent of the composition is larger than 10−10, 10−5, 10−3, 10−1, 0, 1, 5, 10, 50 or 100 kg×K/mol.
The invention also relates to a method for storage or preservation of at least one property of at least one nanoparticle or at least one constituent of the composition such as the chain arrangement, preferentially by following at least one of the following step(s):
wherein the change of state of the state of the composition or at least one constituent of the composition is preferentially selected in the group consisting of: i) liquid to solid, ii) liquid to gas, iii) solid to liquid, iv) solid to gas, v) gas to liquid, and vi) gas to solid.
In one embodiment of the invention, the cryoprotectant or protectant compound occupies a larger volume than the coating or core or nanoparticle or at least one constituent of the composition different from cryoprotectant or protectant compound, preferentially when the composition or at least one constituent of the composition is in a liquid state or is mixed in a liquid.
In one embodiment of the invention, the distance between center of the nanoparticle or at least one constituent of the composition and the external surface of the nanoparticle or at least one constituent of the composition or coating or the thickness of the coating preferentially without the cryoprotectant or protectant compound is smaller than the thickness or diameter of the volume comprising the cryoprotectant or protectant compound.
In one embodiment of the invention, the volume occupied by the coating or nanoparticle or at least one constituent of the composition preferentially without the cryoprotectant or protectant compound is smaller than the volume occupied by the cryoprotectant or protectant compound.
In one embodiment of the invention, the coating material or at least one constituent of the composition has at least one different property from the cryoprotectant or protectant compound, preferentially selected among the group consisting of: i) the coating material is not located in the same region or location than the cryoprotectant or protectant compound in the composition or at least one constituent of the composition, ii) the coating material interacts or complexes more strongly with the nanoparticle core or at least one constituent of the composition core than the cryoprotectant or protectant compound in the composition, and iii) the coating material has a different composition or structure or magnetic property or crystallinity or is different than/from the cryoprotectant or protectant compound or has at least 1, 5, 10, 103 or 105 atom(s) that differ from that(those) of the cryoprotectant or protectant compound.
In one embodiment, the function of the cryoprotectant or protectant compound in the composition or at least one constituent of the composition is to replace water molecule or maintain the size, composition or at least one constituent of the composition, chain arrangement, or magnetic property of the at least one nanoparticle or at least one constituent of the composition, preferentially during cooling or application of a temperature/pressure gradient to the composition or at least one constituent of the composition, preferentially to avoid damages caused by ice on the coating or nanoparticle or at least one constituent of the composition.
In one embodiment, the composition or at least one constituent of the composition is or is maintained at a temperature larger than −273, −50, −1, 0, 2, 5, 10, 100, 103 or 105° C., preferentially for more than 10−10, 0, 1, 5, 10, 103 or 1010 second(s).
In another embodiment, the composition or at least one constituent of the composition is or is maintained at a temperature smaller than 105, 103, 100, 10, 5, 2, 1, 0, −10, −50, −100, −250° C., preferentially for more than 10−10, 0, 1, 5, 10, 103 or 1010 second(s).
In one embodiment, the composition or at least one constituent of the composition is or is maintained under a pressure larger than 10−50, 10−10, 10−5, 10−1, 0, 1, 5, 10, 103, 105, 1010 or 1020 bar, preferentially for more than 10−10, 0, 1, 5, 10, 103 or 1010 second(s).
In another embodiment, the composition or at least one constituent of the composition is or is maintained under a pressure smaller than 1050, 1010, 10, 0, 10−3, 10−5, 10−10 or 10−20 bar, preferentially for more than 10−10, 0, 1, 5, 10, 103 or 1010 second(s).
In one embodiment of the invention, the presence of the cryoprotectant or protectant compound in the composition or at least one constituent of the composition enables to store the composition or at least one constituent of the composition, preferentially in a powder form, preferentially for more than 10−10, 0, 1, 5, 10, 103 or 1010 second(s), preferentially in such a way that the composition or at least one constituent of the composition maintains at least one of its properties.
In one embodiment of the invention, the presence of a nanoparticle or at least one constituent of the composition core, preferentially a metallic one, enhances the effect of the cryoprotectant or protectant compound in the composition or at least one constituent of the composition, preferentially by facilitating the interaction between the coating and the cryoprotectant or protectant compound, preferentially due to a stabilization of the coating on the core.
In one embodiment of the invention, the composition of the or at least one constituent of the composition is a mixture of amorphous and crystalline structure, wherein the nanoparticle or at least one constituent of the composition core is preferentially essentially or dominantly crystalline, wherein the nanoparticle or at least one constituent of the composition coating, cryoprotectant and/or protectant compound is/are preferentially essentially or dominantly amorphous.
In one embodiment of the invention, the cryoprotectant and/or protectant compound protects and/or preserves and/or maintains at least one property of at least one constituent of the composition.
In one embodiment of the invention, at least one constituent of the composition, preferentially amorphous, needs the presence of the cryoprotectant and/or protectant compound to have at least one of its properties protected and/or preserved and/or maintained preferentially following exposure of the composition or at least one constituent of the composition to a temperature and/or pressure gradient(s) and/or radiation, i.e. preferentially without the cryoprotectant and/or protectant compound the at least one property of the at least one constituent of the composition would preferentially be damaged or changed.
In one embodiment of the invention, the at least one constituent of the composition needs the presence of the cryoprotectant and/or protectant compound to have at least one of its properties protected and/or preserved and/or maintained preferentially following exposure of the composition or at least one constituent of the composition to a temperature and/or pressure gradient(s) and/or radiation, i.e. preferentially without the cryoprotectant and/or protectant compound the at least one property of the at least one constituent of the composition would preferentially be damaged or changed.
The invention also relates to a method for the cryo-preservation, size-preservation, composition-preservation, cohesion-preservation, magnetic property-preservation, or preservation of at least one property of the composition or at least one constituent of the composition or nanoparticle by following at least one of the following steps:
In some cases, the preservation of at least one property of the composition or at least one constituent of the composition or nanoparticle can be the variation by less than 105, 103, 102, 90, 75, 60, 50, 25, 10, 5, 2, 1 or 0% of this property, where this variation is preferentially equal to (P2−P1)/P1, where P1 and P2 are preferentially this property before and after the composition or at least one constituent of the composition or nanoparticle have been subjected to a temperature and/or pressure gradient or cooled down or exposed to radiation or oxidized or reduced.
The invention also relates to the composition or at least one constituent of the composition or kit or ensemble or system or assembly comprising:
In some cases, the first and second components or constituents are sold or used separately or at different times or under different conditions.
In some cases, the first component or constituent is necessary or used or serves as starting material for the production or fabrication of the second component.
In some other cases, the first and second components or constituents are sold or used together or at similar times or under similar conditions.
The invention also relates to a method for the cryo-preservation, size-preservation, composition-preservation, cohesion-preservation, magnetic property-preservation, or preservation of at least one property of the composition or at least one constituent of the composition or nanoparticle by following at least one of the following steps:
In some cases, the first cryoprotectant or protectant compound can be used to protect a living organism or can be DMSO or Ethylene glycol or Glycerol or 2-Methyl-2,4-pentanediol (MPD) or Propylene or glycol or Sucrose or Trehalose.
The invention also relates to a method for enabling nanoparticle or at least one constituent of the compositions stored preferentially in powder form with a specific type of assembly or geometric figure preferentially a chain assembly to maintain this type of assembly upon reconstitution preferentially in liquid such as water, where this method preferentially comprises at least one of the following steps of:
The invention also relates to a method of fabrication or cryo-preservation of the composition or at least one constituent of the composition according to the invention and optionally of the at least one cell producing the at least one nanoparticle, which comprises at least one of the following steps:
The invention also relates to a method for storing the composition or at least one constituent of the composition or preserving over time at least one property of the nanoparticle or at least one constituent of the composition according to the invention, or preserving the geometric arrangement or assembly, preferentially chain arrangement of the at least one nanoparticle or at least one constituent of the composition according to the invention, which comprises at least one of the following steps:
The invention also relates to a method for the fabrication, the cryo-preservation, size-preservation, composition-preservation, cohesion-preservation, magnetic property-preservation, or preservation of the composition or at least one constituent of the composition or of at least one property of the composition or at least one constituent of the composition according to the invention, by following at least one of the following steps:
The invention also relates to a method for removing at least one compound from the composition or at least one constituent of the composition according to the invention, preferentially the other compound, by following at least one of the following steps:
The invention also relates to a method for the fabrication, storage, preservation, preservation of the geometric arrangement or assembly, preferentially chain arrangement of the at least one nanoparticle or at least one constituent of the composition, cryo-preservation, size-preservation, composition or at least one constituent of the composition-preservation, cohesion-preservation, magnetic property-preservation, or preservation of at least one property of the composition or at least one constituent of the composition according to the invention, which comprises at least one of the following steps:
In one embodiment, the at least one method or composition or at least one constituent of the composition according to the invention is used for the treatment or detection of a disease or for the preparation or storage or activation or preservation of the composition or at least one constituent of the composition or for the administration of the composition or at least one constituent of the composition to a body part.
In some cases, the composition can be can have at least one common property with the at least one constituent of the composition.
In some other cases, the at least one constituent of the composition is different from or has at least one different property from the composition.
In one embodiment of the invention, the at least one constituent of the composition, preferentially the nanoparticle coating, consists of or comprises at least one compound selected in the group consisting of: i) at least one acid such as citric acid, oleic acid, polymethacrylic acid, poly(ethyleneoxide)-b-poly(methacrylic acid) acid, polyacrylic (PAA) acid, polylactic acid, poly(ethylene oxide)-blockpoly(glutamic acid), Phosphonic acid, ii) Albumin, iii) a bisphosphonate, iv) Alendronate, v) Alginate, vi) a metal, vii) Au, viii) Al2O3, ix) Alginate, x) Aluminium, xi) Aluminium hydroxide, xii) Arabinogalactan, xiii) Bentonite, xiv) cellulose, xv) Carboxymethylcellulose, xvi) Chitosan, xvii) Cholesterol, xviii) Citrate, xix) Dextran, xx) Dimercaptosuccinic acid, xxi) Dopamine, xxii) DOPC, xxiii) DTAP, xxiv) DVB, xxv) Ethylcellulose, xxvi) Erythrocyte, xxvii) Fatty acid, xxviii) Ferrite, xxix) Folic acid, xxx) Gelatin Human, xxxi) serum albumin, xxxii) Liposome, xxxiii) MIPS, xxxiv) MnO, xxxv) Mn3O4, xxxxvi) Oleic acid, xxxvii) PEI, xxxviii) PEG, xxxix) PEO-PGA, xl) PLA (poly(lactide acid), xli) PLGA, xlii) Phosphatidylcholine, xliii) Phosphorylcholine, xliv) Pluronic, xlv) Polyacrylamide, xlvi) Polyacrylic acid, xlvii) PAA, xlviii) Polyaniline Polyethylene glycol preferentially with terminal carboxyl groups, xlix) peptides, L) polypeptides, Li) Poly(ethylene oxide), Lii) Poly(vinyl alcohol), Liii) Ploy(N-isopropylacrylamide), 54) Poly(vinylpyrrolidone), 55) Poly(oligoethylene oxide), 56) Poly(N,N-dimethyl ethylamino acrylate), 57) Poly(imine), 58) Poly(acrylic acid), 59) Poly-D-L lactide, 60) Polyalkylcyanoacrylate, 61) Polymer, 62) PAMAM or PDMAEMA or PPEGMA or PolyNIPAAM, 63) Polyacrylic acid, 64) Polydipyrrole/dicarbazole, 65) Poly-L-lysine, 66) Polymethylmethaacrylate, 67) Polymersomes, 68) Polysaccharide, 69) Agarose or alginate or carrageenan or chitosan or dextran or heparin or Gum Arabic or Pullulan or Starch, 70) Polystyrene, 71) PVA, 72) PVP Silica, 73) an amorphous or mesoporous compound, 74) Silane, 75) SiO2, 76) Sodium Oleate, 77) Starch, 78) Styrene, 79) TaOx, 80), ZrO2, 81) Polysacharides, 82) Agarose or alginate or carrageenan or chitosan or dextran or carboxy-methyl-dextran or heparin or Gum Arabic or Pullulan or Starch, 82) Acide, 83) polymethacrylic acid or poly(ethyleneoxide)-b-poly(methacrylic acid) or polyacrylic acid (PAA) or polylactic acid or poly(ethylene oxide)-blockpoly(glutamic acid) or Phosphonic acid or Dimercaptosuccinic acid or Fatty acids or folic acid or PLA (poly(lactide acid) or Polyacrylic acid PAA 84) Polymer, 85) Dextran or Poly(ethylene oxide) or Poly(vinyl alcohol) or Ploy(N-isopropylacrylamide) or Poly(vinylpyrrolidone) or Poly(oligoethylene oxide) or Poly(N,N-dimethyl ethylamino acrylate) or Poly(imine) or Poly(acrylic acid), 86) Carboxylate, 87) inorganic compound, 88) SiO2 or Al2O3 or ZrO2 or ferrite or MnO or Mn3O4 or Au or Bentonite or Carbon preferentially activated, graphitized, 89) organic metals, 90) MIPs or Celulose or DV8 or Ppy or Chitosan or Polyacrylamide or alginate or PEI or surfactants or Phosphates or Silica or Gold or Dextran or PEG or Alginate or Chitosan, 90) or compound with a chemical function such as Alcohol for example PVA (poly(vinyl alcohol), amide, for example Poly(N-isopropylacrylamide), aldehydes, 91) a compound with a type of interactions with avec hydroxyl groups at the surface of the iron oxide (Fe—OH), 92) a compound with electrostatic interactions, preferentially with a difference in charge between at least two constituents of the composition or at least one constituent of the composition, 93) a compound with a hydrophobic, chelating, covalent, interaction or bond, 93) a compound with a type of functional group that can preferentially be attached at the surface of a metal or iron oxide such as —OH, 94) PEG or Dextran or Polyvinylalcool or pluronic or Dopamine or Amine or Cysteine or phosphonic acid or carboxylic acid or trimethoxy or silane, 95) a compound with a —NH2 group, 96) Chitosan or polyethylenimine or Ploy(L-lysine) or PEG with terminal amine groups or Ethylenamine, 97) a compound with a —COOH group, 98) Polyacrylic acid, 99) carboxymethylcellulose, 100) PEG with terminal carboxyl groups, 101) alginate, 102) Polymethacrylic acid, 103) citrate, and 104) iminodiacetic acid or nitrilotriacetic acid or ethylendiamin or tetraacetic acid or diethylenetrianine pentaacetic acid or folic acid or L-cysteine or an amino acid or Thiol or Dimercaptosuccinic acid or a Phosphate compound or pyridoxal phosphate or adenosine diphosphate or nicotinamide adenine dinucleotide phosphate.
The invention also relates to a method for activating the composition or at least one constituent of the composition or at least one center of activity of the composition or at least one constituent of the composition, according to the invention, preferentially by applying a radiation or physico-chemical disturbance on the composition or at least one constituent of the composition preferentially for a certain or sufficiently long time, preferentially for more than 10−3, 1, 0, 1, 10, 103 second(s),
In one embodiment of the invention, the chelating agent or the at least one constituent of the composition is selected in the group consisting of: (i) chelating agents which have one or more carboxyl groups, (ii) chelating agents which have one or more hydroxyl groups, (iii) chelating agents which have one or more amino and/or carboxyl and/or ketone groups, (iv) chelating agents which have one or more phosphonate and/or phosphonic acid groups, (V) chelating agents which have one or more bisphosphonate and/or trisphosphonate and/or tetra phosphonate groups, (vi) chelating agents which have one or more Sulfonate and/or Sulfonic acid groups, and (vii) chelating agents of monodentate or polydentate type or poly chelating agent that may comprise one or a plurality of functional groups, for example among carboxyl, hydroxyl or amino groups, (viii) of polysaccharide type. (ix) the chelating agents with one or more carboxyl groups or 9) ALA (alpha-lipoic acid) or 10) calcein or 11) carboxyfluorescein or deferasirox or dipicolinic acid or DTPA (diethylenetriaminepen taacetic acid) or EDTA (ethylenediaminetetraacetic acid) or folic acid or vitamin B9 or lactic acid or rhodamine B or carboxymethyl dextran or oxalic acid or citric acid or a compound comprising one or more citric and/or citrate functional groups or phenolic acid or 12) a chelating agent comprising one or more acetate and/or acetic functional groups or BAPTA (aminophenoxyethanetetraacetic acid) or CDTA (cyclohexane-1,2-diaminetetraacetic acid) or EDDHMA (ethylenediaminedi(ohydroxy-p-methylphenyl)acetic acid) or CaNa-EDTA or EDTCA (ethylenediaminetetraacetic acid with Cetavlon® or ammonium-type surfactant or EDDA or ethylenediamine-N,N′-diacetic acid) or EDDHA or ethylenediamine-N,N′-bis(2-hydroxyphenylacetic acid or EGTA or ethylene glycol bis(B-aminoethyl ether)-N,N,N′,N′-tetraacetic acid or HEDTA (N-(2-hydroxyethyl)ethylenediaminetriacetic acid or HEEDTA or hydroxy-2-ethylenediaminetriacetic acid or NTA or nitrile triacetate or 13) a molecule comprising one or more hydroxyl functional groups, such as catechol or derivatives thereof, or else deferiprone or 14) a molecule comprising one or more amino functional groups, such as dopamine and/or deferoxamine or 15) a molecule comprising one or more aminocarboxylic and/or ketone functional groups, such as doxorubicin, caffeine, D-penicil lamine, pyrroloquinoline and HEIDA (hydroxyethylimino N,N-diethanoic acid) or 16) a molecule comprising at least one phosphonate or phosphonic functional group, such as AEPN (2-aminoethylphosphonic acid) or AMP (aminotris(methylenephosphonate)) or ATMP (aminotris(methylenephosphonic acid)), CEPA (2-carboxy ethylphosphonic acid) or DMMP (dimethyl methylphosphonate) or DTPMP (diethylenetriaminepenta(methylenephos phonic acid)) or EDTMP (ethylenediaminetetra (methylenephosphonic acid)) or HEDP (1-hydroxyethylidene 1,1-diphosphonic acid) or HDTMP (hexamethylenediaminetetra(methylenephosphonic acid)) or HPAA (2-hydroxyphosphonocarboxylic acid) or PBTC (phosphonobutanetricarboxylic acid), PMIDA (N-(phosphor nomethyl)iminodiacetic acid) or TDTMP (tetramethylenediamine-tetra(methylenephosphonic acid)) or ADP (adenosine diphosphoric acid) or 1-12-4-(dipyrromethene boron difluoride)butanoyl)aminododecanoyl-2-hydroxy-sn-glycero-3-phosphate or a sodium salt of L-O-phosphatidic acid or a sodium salt of 1-palmitoyl-2-(dipyrromethene boron difluoride)undecanoyl-sn-glycero-3-phospho-L-serine or 17) a molecule containing at least one bisphosphonate, trisphosphonate or tetraphosphonate functional group, such as 1-hydroxymethylene-bis-phosphonic acid, propanetriphosphonic acid, (nitrilotris(methylene))trisphosphonic acid or (phosphinylidynetris(methylene))trisphosphonic, or 18) a molecule comprising one or more Sulfonate or Sulfonic acid functional groups, or else a dimercapto group, such as BPDS (bathophenanthroline disulfonate or 4,7-di(4-phenylsulfonate)-1,10-phenanthroline), DMPS (dimercaptopropane Sulfonate or 2,3-dimercapto-1-propanesulfonic acid), Sulforhodamine 101 or DMSA (dimercaptosuccinic acid) or 19) polydentate ligands, i.e. chelating agents having more than one atom capable of binding to a metal atom, such as hemoglobin, chlorophyll, porphyrins and organic compounds containing pyrrole rings or 20) polymeric compounds, in particular polysaccharide compounds or 21) rhodamine B, ascorbic acid, citric acid, folic acid, erythrosine, hemoglobin, a low-molecular-weight dextran, anthranilic acid, calcein, alendronate, 3-cyclohexy lamino-1-propanesulfonic acid (CAPS) or EDTA.
The invention also relates to the composition or at least one constituent of the composition according to the invention, wherein the composition or at least one constituent of the composition, comprises an organic part and/or an inorganic part, wherein the inorganic part preferentially comprises the core of the nanoparticle or at least one constituent of the composition, wherein the organic part preferentially comprises the coating of the nanoparticle or at least one constituent of the composition and/or the cryoprotectant or protectant compound, and wherein the percentage in mass of the inorganic part is preferentially larger than the percentage in mass of the organic part.
In one embodiment of the invention, a protectant compound is a compound that maintains or prevents the variation of at least one property of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition preferentially by more than 1020, 1010, 105, 10, 5, 2, 10% or preferentially by a factor of more 1020, 1010, 105, 10, 5, 2, 10, preferentially over time, preferentially over more than 1 second or 1 year, preferentially between the time t1 and t2, where the properties of the nanoparticle or at least one constituent of the composition or composition or at least one constituent of the composition at times t1 and t2 are preferentially P1 and P2.
In some cases, P2/P1 and/or t2/t1 can be smaller than 1020, 1010, 105, 10, 5, 2, 10. In some other cases, P2/P1 and/or t2/t1 can be larger than 0, 10−10, 0.1, 0, 1, 10, 50, 103 or 105.
In some cases, the factor α is equal to V2/V1, where V2 is the volume occupied by the cryoprotectant or protectant compound in the composition or at least one constituent of the composition and V1 is the volume occupied by the at the at least one nanoparticle or at least one constituent of the composition or the at least one chain in the composition or at least one constituent of the composition.
In some cases, α is larger than or equal to 1, 2, 5, 10, 100, 103 or 105.
In some other cases, α is smaller than or equal to 105, 103, 100, 10, 5, 2 or 1.
In some other cases, α is larger when the composition or at least one constituent of the composition is partly or fully in a liquid state than when it is partly or fully in a solid or powder form.
In some cases, the percentage in mass of cryoprotectant or protectant compound or nanoparticle or at least one constituent of the composition is larger than or equal to 0, 10−10, 10−1, 1, 5, 10, 50, 75, 99 or 100%.
In some other cases, the percentage in mass of cryoprotectant or protectant compound or nanoparticle or at least one constituent of the composition is smaller than or equal to 100, 99.99, 99, 85, 75, 50, 25, 20, 10, 5, 2, 1 or 0%.
In still some other cases, the percentage in mass of the nanoparticle or at least a first constituent of the composition is larger preferentially by a factor β than the percentage in mass of the cryoprotectant or protectant compound or of at least one second constituent in or of the composition, preferentially when the composition or at least one constituent of the composition is in powder or solid form.
In some cases, the factor β is equal to PM2/PM1, where PM2 is the percentage in mass of the at least one nanoparticle or at least a first constituent of the composition and PM1 is the percentage in mass of the cryoprotectant or protectant compound or at least one second constituent of the composition.
In some cases, β is larger than or equal to 1, 2, 5, 10, 100, 103 or 105.
In some other cases, β is smaller than or equal to 105, 103, 100, 10, 5, 2 or 1.
In one embodiment of the invention, the nanoparticle or nanoparticle core or least one constituent of the composition comprises a percentage in mass in at least one metal preferentially iron preferentially in terms of metallic composition that is larger than 0, 1, 5, 10, 50, 75, 90, 99 or 99.9%.
In one embodiment of the invention, the nanoparticle or nanoparticle coating or least one constituent of the composition comprises a percentage in mass in at least one metal preferentially iron preferentially in terms of metallic composition that is smaller than 100, 99.9, 90, 75, 50, 25, 10, 5, 2 or 1%.
In one embodiment of the invention, the percentage(s) in mass of the nanoparticle or protectant compound, or at least one constituent of the composition is(are) larger than or equal to 0, 1, 5, 10, 50, 75, 90, 99 or 99.9%.
In another embodiment of the invention, the percentage(s) in mass of the nanoparticle or protectant compound, or at least one constituent of the composition, is(are) smaller than or equal to 100, 99.9, 90, 75, 50, 25, 10, 5, 2 or 1%.
In one embodiment of the invention, the percentage(s) in mass of the inorganic and/or organic part(s) in the composition or at least one constituent of the composition, preferentially in the dried composition is(are) larger than or equal to 0, 1, 5, 10, 50, 75, 90, 99 or 99.9%.
In another embodiment of the invention, the percentage(s) in mass of the inorganic and/or organic part(s) in the composition or at least one constituent of the composition, preferentially in the dried composition, is(are) smaller than or equal to 100, 99.9, 90, 75, 50, 25, 10, 5, 2 or 1%.
In one embodiment of the invention, the nanoparticle or at least one constituent of the composition, has/have at least one of the following properties selected in the group consisting of:
In some cases, the coating and core form a complex.
In some other cases, the cryoprotectant or protectant compound and nanoparticle or at least one constituent of the composition don't form a complex or form a weaker or less stable complex than the complex formed by the coating and core.
In one embodiment of the invention, the at least one method of the invention is used for at least one of the following purposes: i) decreasing the production of free radicals, ii) reducing radiation, iii) reducing the effect of radiation, iv) increasing the destruction of the body part, preferentially pathological or tumor cells or a tumor or cancer or virus or a pathological part of the body part, v) destroying or inactivating at least 1, 1.1, 2, 5 or 10 times more pathological or tumor cells or virus preferentially when the pathological cell or pathological body part is exposed to radiation in the presence of the composition than when the pathological cell or pathological body part is exposed to radiation in the absence of the composition, vi) reducing side effects of radiation or preserving the body part, preferentially non-pathological or healthy cells or healthy body part, preferentially surrounding the pathological body part, vii) preserving or maintaining activated or alive at least 1, 1.1, 2, 5 or 10 times more healthy cells preferentially when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition than when the healthy cells are exposed to radiation or subjected to an indirect such as an immune reaction that occurs after the application of radiation on the body part in the presence of the composition in the absence of the composition, viii) sonodynamic therapy, ix) photodynamic therapy, x) radiation therapy, xi) a diagnosis, and x) a treatment.
In one embodiment of the invention, at least one method according to the invention comprises at least 1, 2, 3 or 4 step(s), which is repeated at least 1, 2, 3, 4, 5 or 10 time(s).
In another embodiment of the invention, at least one method according to the invention comprises at least 1, 2, 3 or 4 step(s), which is repeated less than 10, 5, 2 or 1 time(s).
In some cases, CA1 can be C1FRP or have at least one property in common with C1FRP.
In some other cases, CA2 can be C2FRP or have at least one property in common with C2FRP In some cases, the constituent of the composition can be selected in the group consisting of: i) the constituent of the composition, ii) the other constituent of the composition, iii) the first constituent of the composition, and v) the second constituent of the composition.
The description is followed by the following non-limiting example(s):
We report a method to formulate natural iron oxide nanoparticle or at least one constituent of the compositions, called magnetosomes, by amplifying magnetotactic bacteria in non-toxic growth media, by extracting these nanoparticle or at least one constituent of the compositions from magnetotactic bacteria under alkaline lysis, by purifying them by heat above 400° C. This results in pure non-pyrogenic magnetosome minerals, M-uncoated, which are further coated with biocompatible citric acid or carboxy-methyl-dextran compounds, to yield stable M-CA and M-CMD. The last steps of the formulation consist in adding sorbitol, preferentially 5% sorbitol, to M-CMD and a mixture of sucrose and PEG 4000, preferentially 3.75% sucrose and 1.25% PEG 4000, to M-CA, in lyophilizing these mixtures, resulting in NP powders of (M-CMD)f and (M-CA)f that display a long term stability, preferentially over at least 6 months, having preserved their pre-lyophilization properties, i.e. their stability in suspension, their chain arrangement, their carbon content, as well as their surface charge and surface chemical groups. in addition, we have established that (M-CMD)f and (M-CA)f are isotonic, preferentially with osmolalities comprised between 275 and 290 mosm/kg H2O upon NP reconstitution in water, fully biocompatible, i.e. sterile, non-pyrogenic, non-cytotoxic towards 3T3, L929, and V79 healthy cells up to a NP concentration of 1 mg/ml, and efficient in destroying prostate PC3-luc tumor cells when they are heated up to a maximum temperature of 46° C. during 30 minutes in the presence of these cells under the application of low intensity ultrasound or alternating magnetic field. These results demonstrate that (M-CMD)f and (M-CA)f display a long-term storage capacity, a full biocompatibility, as well as a potential to destroy tumor cells under hyperthermia.
Among the different types of nanoparticle or at least one constituent of the compositions (NP), those made of iron oxide (IONP) display one of the highest level of biocompatibility, hence enabling their use for medical applications, in particular cancer treatment. Chemically synthesized IONP often suffer from a series of drawbacks such the use of toxic compounds in their fabrication process, a crystallinity, non-uniform shapes, and/or small sizes with low magnetization. To overcome these disadvantages, a biological synthesis route has been developed, in which IONP, called magnetosomes, are produced intracellularly by gram-negative magnetotactic bacteria (MTB). MTB have adjusted magnetosome properties over millions of years through a Darwinian process to yield an optimized system of magnetic guidance also called magnetotactisme in which the magnetosome magnetic moment aligns parallel to the earth magnetic field, enabling MTB to orientate in this direction, therefore easing their search for an optimal living environment. Magnetosomes thus consist of magnetite (Fe3O4) well crystalized cuboctahedric nanocrystals surrounded by a stabilizing phospholipid bilayer originating from MTB. They display at individual level a size of 35 to 120 nm, leading to ferrimagnetic properties.
At the larger scale of magnetosome assembly, they are observed to for chains, which prevent their agglomeration. While these features led to superior antitumor efficacy for magnetosomes than for their chemical counterparts, such results were obtained with magnetosomes in non-finalized formulations.
Part of our work consists in improving the magnetosome fabrication process to render these NP injectable to humans For that, toxic CMR or animal-based compounds are first removed from MTB growth media. second, the natural organic magnetosome membrane that contains lipopolysaccharides known as endotoxins is removed and further treatments are undertaken to yield magnetosome minerals designated as M-uncoated, which are non-pyrogenic but prone to agglomeration/sedimentation due to strong magnetic dipole interactions. Third, M-uncoated are covered with biocompatible compounds trough coordinate bonds between a coating material, i.e. citric acid (CA) or carboxy-methyl dextran (CMD), and iron cations at mineral surface, to yield stabilized M-CA or M-CMD. The further steps, which are the topic of this example, consist in adding cryoprotectants to M-CA and M-CMD and lyophilizing the obtained mixtures to transform them in a NP powder, designated as (M-CA)f and (M-CMD)f, which can be stored for a long period of time without losing its properties.
To improve long-term storage stability and avoid disassembling the coating from the magnetosome minerals, coated magnetosome minerals should better be stored in powder than liquid suspension. To eliminate water from M-CA and M-CMD, we have lyophilized these NP in the presence of various cryoprotectants, whose cryoprotecting action possibly relies on the creation of an amorphous glassy matrix that immobilizes nanoparticle or at least one constituent of the compositions during freezing and therefore avoids that crystallized ice produced during this step damages the coating. Since the efficacy of cryoprotection presumably depends on the type and concentration of cryoprotectant used in a formulation, we have tested various cryoprotectants, i.e. Glucose, Mannitol, PEG 4000, Sorbitol, Sucrose, Trehalose, at different concentrations, which we have added to M-CA and M-CMD. We have then determined the optimized conditions enabling to obtain stable and isotonic (M-CA)f and (M-CMD)f, which can be stored in powder form and resuspended in water, while maintaining their physico-chemical properties, i.e. their coating thickness/composition or at least one constituent of the composition or magnetosome chain arrangement, their biocompatibility, i.e. their sterility, non-pyrogenicity, non-cytotoxicity towards healthy cells, as well as their faculty to destroy PC3-luc tumor cells under mile hyperthermia conditions of 41-46° C. triggered by the application of low intensity ultrasounds or alternating magnetic field.
Materials: Magnetospirillum gryphiswaldense strain MSR-1 (DSM 6361) was purchased from Deutsche sammlung von mikro-organismen and zellkulturen (brunswick, germany). Pc3-luc tumor cell line derived from PC3 human prostate adenocarcinoma was purchased from Perkinelmer. Mouse fibroblast 3T3 cell line was purchased from American type culture collection (ATCC® CCL-163™) Hamster Chinese lung male V79-4 cell line was purchased from ATCC® (CCL-93). Mouse fibroblast L929 cell line was purchased from ATCC® (CCL-1). Potassium hydroxide (KOH) and citric acid were purchased in pharmaceutical grade from Merck. Carboxymethyl dextran (CMD) was purchased from TDB labs. Sucrose, glucose, trehalose, mannitol, sorbitol and polyethylene glycol 4000 (peg 4000) were purchased in pharmaceutical grade from Merck. Dextran T1 was purchased from pharmacosmos. Tryptic soy broth (TSB) and fluid thioglycolate medium (FTM) were purchased from Merck. Phosphate-buffered saline (PBS), sodium hydroxide (NOH), hydrochloric acid (HCl), nitric acid (HNO3), and Triton X-100 were purchased from Thermo fisher scientific. Dulbecco's modified eagle's medium (DMEM) with/without phenol red, fetal bovine serum (1-BS), penicillin-streptomycin, HEPES buffer solution, 0.25% trypsin—EDTA solution, and trypan blue solution were purchased from Gibco. Resazurin dye was purchased from Invitrogen.
Culture of MSR-1 magnetotactic bacteria: Briefly, MSR-1 bacteria were first amplified in two steps of pre-culture (PC) with the iron-deficient pre-growth medium, then cultivated in the last step with the appropriate conditions to promote the synthesis of magnetosomes. During the PC 1, 300 μl of MSR-1 cryo-stock were incubated in 50 ml of pre-growth medium at 29.5° C. for 6 days in static state followed by 1 day under agitation at 110 rpm. Then, the pc1 was performed by transferring all the cells in a 151 bioreactor (Applikon) containing 61 of pre-growth medium and agitated at 200 rpm and 29.5° C. for 3 days. during the last step of culture, the PC 2 cells were transferred in a 40 L bioreactor (Applikon) filled with 26 l of growth medium. The culture conditions such as oxygen concentration, temperature and agitation speed were respectively maintained between 0.1 and 1%, at 29.5° C. and under 200 rpm during the entire fermentation. the pH was kept at 6.9 by automatically adding an acidic medium (pH=3), called the “fed-batch” solution. The bacterial growth was followed at every 24 h by taking a sample of 5 ml from the culture to determine the optical density at 565 nm (OD565) using a UV-visible spectrophotometer. MSR-1 magnetotactic bacteria (MTB) were cultivated for a total of 19 days, using bacterial growth media composed of pharmaceutical grade chemicals, which are free of toxic compounds and heavy metals other than iron. The pre-growth medium (PGM), growth medium (GM) and fed-batch medium (1-BM) comprise: i) sodium lactate (preferentially 2.6 g/L in PGM, 1.3 g/L in GM, 100 g/L in 1-BM), ii) ammonium chloride (preferentially 0.4 g/L in PGM, 0.2 g/L in GM, 4.8 g/L in FBM), iii) magnesium sulfate heptahydrate (preferentially 0.1 g/L in PGM, 0.03 g/L in GM, 2.4 g/L in 1-BM), dipotassium phosphate (preferentially 0.5 g/L in PGM, 0.07 g/L in GM, 6 g/L in FBM), iron (III) chloride hexahydrate (preferentially 0 g/L in PGM, 0 g/L in GM, 2 g/L in FBM), mineral elixir (preferentially 0.5 mL in PGM, 0.08 mL in FBM, 7 mL in 1-BM), vitamin elixir (preferentially 0.1 mL in PGM, 0.07 mL in GM, 1 mL in FBM). The mineral elixir comprises 1 g/L of iron (II) sulfate heptahydrate and 30 g/L of calcium chloride. The vitamin mixture comprises 0.002 g/L biotin, 0.4 g/L calcium penthotenate, 0.002 g/L folic acid, 2 g/L inositol, 0.4 g/L nicotinic acid, 0.2 g/L p-aminobezoic acid, 0.4 g/L pyridoxine HCl, 0.2 g/L riboflavin, 0.4 g/L Thiamine HCl. The 10 first days include 2 steps of pre-culture used to amplify the bacteria in the pre-growth medium devoid of the iron source. The third step lasts about 9 days. it consists in promoting the intracellular synthesis of magnetosomes by cultivating MSR-1 bacteria in their optimized microaerophilic conditions and by gradually adding iron ion sources via fed-batch solution. At the end of the bacterial culture, 32 l of bacterial culture were concentrated to reduce the liquid volume using a tangential filter column. After that, MSR-1 concentrate (about 51 in total) was stored at −80° C. for further steps of magnetosome production.
Extraction and purification of magnetosomes: Briefly, the concentrated MSR-1 cells were diluted at an OD565 value of 20 and then lysed in 2 m KOH at 80° C. for 1 h under mechanical stirring at 150 rpm. Using magnetic selection, magnetosome chains (MgC) were collected and washed 2 times with 1× phosphate-buffered saline (PBS) followed by 3 times washing with deionized water. After being concentrated in 50 nil-conical tubes, Mgc were frozen at −80° C. for 24 h and next dried at −50° C., 0.003 mbar for 24 h using a lyophilizer (labconco, free zone 70020 2.5 l) to transform them into powder. Subsequently, 100 mg of this powder were heated at 6° C./min in a muffle furnace using a heating program comprised of several heating steps of temperatures comprised between 50° C. and 420° C. (maintained for 2 h), for purifying the organic material of post-extracted magnetosomes and yielding magnetosome minerals or “uncoated magnetosomes” (M-uncoated).
Coating of M-Uncoated with citric acid (CA) and carboxymethyl-dextran (CMD): The coating process was carried out under aseptic conditions, using a biosafety hood and pyrogen-free materials. The solutions of CA and CMD were prepared in pyrogen-free water at 25 mg/ml and 150 mg/ml, respectively. These solutions were then filtered with a 0.22 μm polyethersulfone filter for sterilization. After that, 200 ml of each solution (CA or CMD) were added to the 1 l-glass beaker containing 2 g of M-uncoated (corresponding to 1 g of Fe) and 300 nil of pyrogen-free water. Each suspension was then sonicated for 1 min in pulse mode with a pulse length of 0.1 s and pulse interval of 0.1 s, at 20 W and ambient temperature, using a probe sonicator (PS) (branson, digital sonifier s-250d) with 25 mm-diameter tips. following the short time sonication, the pH values were adjusted at 6 and 4.5 for CA and CMD, respectively, using 1 M NaOH or 1 M Hcl. Subsequently, these two suspensions were re-sonicated for 1 h using the PS (Branson, digital sonifier s-250d) at the same parameters as described earlier to prepare magnetosome minerals coated with CA and CMD, called M-CA and M-CMD respectively. After the sonication, the suspensions of M-CA and M-CMD were centrifuged at 10° C. and 3380 g for 45 min to remove excess of coating agents in the supernatant. each coated magnetosome minerals were then re-suspended in 10 nil of water to obtain a final concentration of 100 mg/ml of iron and stored at +4° C.
Selection of cryoprotectants: Under aseptic conditions, 0.5 nil of M-CMD suspension at 100 mg/ml of iron was mixed with 0.5 ml of various cryoprotectant solutions, such as glucose, mannitol, sorbitol, sucrose, and trehalose, to obtain the formulated samples of 1 ml at 50 mg/ml of iron containing 5% or 10% w/v cryoprotectant. each sample was immediately frozen in nitrogen liquid for 15 min after 30 s of vortexing, then lyophilized at −50° C. (shelf temperature), 0.003 mbar during 20 h for primary desiccation (labconco, free zone 70020 2.5 l). Afterward, the secondary desiccation during 6 h was triggered by increasing the temperature to 40° C. (shelf temperature) with a heating rate of 0.3° C./min while maintaining the pressure at 0.003 mbar. Likewise, M-CA suspension at 100 mg/ml of iron tested with different cryoprotectant solutions to obtain the formulated samples of 1 ml at 50 mg/ml of iron containing 5% or 10% w/v cryoprotectant for glucose, mannitol, sorbitol, trehalose, and sucrose; 0.5%, 2.5%, 5%, 7.5%, 10% and 15% w/v cryoprotectant for peg 4000 and dextran t1; and 2.5%, 3.75%, 5%, 6.25% and 7.5% w/v cryoprotectant for the combinations of sucrose—peg 4000 and sucrose—Dextran T1 with the mass ratios of sucrose to PEG or Dextran of 1:1, 1:2, 1:3, 2:1, 2:2, 2:3, 3:1, 3:2 and 3:3. After lyophilization, some formulated magnetosome samples were resuspended in 1 ml of sterile water for further characterizations. The selected formulations of M-CA and M-CMD with cryoprotectants were called (M-CA)f and (M-CMD)f, respectively.
Transmission electron microscopy (TEM): 1 ml of MSR-1 bacteria at the end of the culture was diluted at an OD565 nm, value of 1 and then rinsed 2 times with deionized water by centrifugation at 2400 g for 10 min. The different magnetosome suspensions were diluted at 50 μg/ml of iron. Afterward, 7 μl of each sample were deposited on a carbon-coated copper grid (300 mesh from oxford instruments). The grid was then left dry at ambient temperature for at least 3 h before being observed under a transmission electron microscope (Jeol JEM-2100) operated at 200 kv. Using the image j software, nanoparticle or at least one constituent of the composition size was estimated by measuring the diameter of around 400 mineral cores randomly selected from magnetosome chains inside the MSR-1 bacteria.
Colloidal stability by colorimetric dosage: 1 ml of magnetosome suspension was vortexed for 30 s then a sample of 50 μl was immediately taken, called “the sample t0”. the suspension was then rested on the bench for 2 hours and another sample of 50 μl was subsequently taken from the superior part of the liquid, called “the sample t2h”. To transform into iron ions, each sample was dissolved overnight in 950 μl of 37% v/v HCl at ambient temperature. Then 20 μl were withdrawn and mixed with 50 μl of 20% v/v H2O2 for 15 min to oxidize Fe2+ to Fe3+ ions. After that, the solution was added with 880 μl of ultrapure water and then mixed with 50 μl of 2 m KSCN to form the complex between fe3+ ions and thiocyanate ions, whose absorption at 476 nm was measured in a spectrophotometer. The concentration in iron was consequently determined using a calibration range. the stability rate of magnetosome suspension was calculated as the ratio of the concentration in iron of the sample t2h to that of the sample to.
Osmolality measurement: 30 μl of magnetosome suspension at 50 mg/ml of iron were introduced into a micro-sample tube and then placed on the plate of the osmometer (advanced osmometer, model 2020) to determine the osmolality of each sample.
CHNS elemental analysis: 5 mg of magnetosome powder were packed in an aluminum capsule and introduced into CHNS elemental analyzer (thermofisher, flash 2000) to determine the rate of carbon and nitrogen from organic materials in each preparation.
Fourier transform infrared spectroscopy (FT-IR): 2 mg of magnetosome powder was deposited on a germanium crystal plate, connected to an FT-IR spectrometer. The tip was lowered until contact with the crystal and thus the powder sample. FT-IR spectra were then recorded in a scanning range of 590-4000 cm−1 with a resolution of 4 cm−1.
Zeta-potential measurement: Each magnetosome suspension was diluted at 50 μg/ml in iron with ultrapure water. Then it was divided into five samples of 2 ml for preparation at different pH varied from 2 to 10 at 25° C. using HCl and NaOH solutions. 1.5 nil of each sample was subsequently introduced in a 4.5 nil-disposable cuvette and measured in a zeta potential analyze.
Residual moisture analysis: 10 mg of lyophilized powder were placed in an aluminum oxide crucible and introduced into thermogravimetric analysis (TGA) instrument. Then under the flow of nitrogen gas, the sample was heated from 30° C. to 120° C. at 6° C./min and maintained at 120° C. for 1 h to determine the weight loss of lyophilized powder.
Sterility test: Under aseptic conditions, 20 mg of each magnetosome powder were incubated in two conditions, 10 ml of tryptic soy broth (TSB) at 25° C. and 10 nil of fluid thioglycolate medium (FTM) at 35° C. for 14 days. the culture media without NP were considered negative controls. on the last day, 1 ml was taken from each sample and placed against the magnet for 10 min to only recuperate the medium. The turbidity was then measured with 900 μl of such medium at 600 nm. And the remaining 100 μl were incubated on a solid Luria-Bertani agar plate at 25° C. or 35° C. for 3 days to possibly detect contaminant colonies. The other tests, i.e. growth promotion test and method suitability test, were also carried out in parallel with the sterility test of magnetosomes to verify the robustness of this test.
Endotoxin quantification by the limulus amebocyte lysate (LAL) assay: Endotoxin quantification was carried out on different magnetosome powders using a “pierce chromogenic endotoxin quant” kit under aseptic conditions. All materials contacting NP samples were sterile and pyrogen-free. Before the assay, each magnetosome powder was prepared at 40 μg/ml in iron in endotoxin-free water and then heated at 70° C. for 15 min to denature any residual proteins interfering with the assay. After that, 25 μl from such suspension was introduced into a 96 well-plate pre-equilibrated at 37° C. for at least 10 min. Keeping the plate at 37° C., 25 μl of the reconstituted amebocyte lysate reagent were added to each well and incubated for 12 min, followed by the incubation with 50 μl of the reconstituted chromogenic substrate for 6 min. Ultimately, 25 μl of 25% acetic acid was added to each well to stop the reaction. The optical density of such mixture was subsequently measured at 405 nm in a microplate spectrophotometer. The concentration of endotoxin was determined by the standard curve of E. coli endotoxin which was established at the same time as the samples.
Growth of various cell lines: 3T3, L929, V-79, PC3-luc: A 2 ml-cryotube of different cell lines (3T3, L929, V-79, or PC3-Luc) was thawed in a water bath at 37° C. for 10 min Each of these cell lines was then introduced into a 75 cm2-culture flask containing 10 ml of suitable culture medium, i.e. DMEM supplemented with 10% v/v of fbs, 1% v/v of penicillin-streptomycin mixture used for 3T3, V-79, and PC3-Luc, while DMEM supplemented with 10% v/v of hs, 1% v/v of penicillin-streptomycin mixture, and 1% v/v of HEPES buffer for L929. After being sed in the flask, the cells were maintained at 37° C. in a 5% CO2 incubator. The culture medium was renovelled twice per week for each flask. when the cells reached about 80% confluence, the cell passage was carried out. For that, all the liquid medium was first removed from the flask, and 1 nil of 0.25% trypsin—EDTA was added to collect all the adherent cells in suspension. then only 0.5 ml of cell suspension was retained to be mixed with 10 ml of fresh medium in a novel 75 cm2-culture flask and re-incubated at 37° C. in the 5% CO2 incubator. Following 3 passages, each cell line was ready for further experiment.
Cytotoxicity test. 3T3, L929, and V-79 cell lines were each seeded on a sterile 96-well plate (104 cells/well) and then incubated overnight at 37° C. in a 5% CO2 incubator for cell adhesion. After that, all the medium was delicately removed from the plate. 100 μl of the suspension of lyophilized (M-CA)f or (M-CMD)f, prepared at various concentrations in the suitable culture medium without phenol red for each cell line, were then added to each well. In 3T3, V-79, L929 cell plate, 0.001, 0.1, 0.25, and 1 mg/ml in iron of magnetosome suspensions were added. After adding the nanoparticle or at least one constituent of the composition suspensions, the plates were re-incubated for 24 h at 37° C. in a 5% CO2 incubator. To determine the cell viability, 10 μl of resazurin reagent were added to each well and homogenized under agitation at 120 rpm for 10 min. The plates were next incubated at 37° C. in the 5% CO2 incubator for approximately 4 h. Once the blue color in the wells of untreated cells turned to pink color, all the liquid in each well was transferred to a 1.5-ml Eppendorf tube. The tube was then centrifuged at 14100 g for 10 min to remove all the nanoparticle or at least one constituent of the compositions which could interfere with the assay. Subsequently, 100 μl of the supernatant was transferred into a novel 96 well-plate to be measured at excitation and emission wavelengths of 530 and 590 nm, respectively, using a fluorescence reader. The cell viability was calculated using the formula:
where, Ft is the fluorescence intensity of treated wells, Fb is the fluorescence intensity of blank measured in the empty wells containing 100 μl of medium mixed with 10 μl of resazurin, and Fc is the fluorescence intensity of control wells measured in the untreated cells incubated in 100 μl of medium mixed with 10 μl of resazurin.
Hyperthermia treatments under AMF (alternating magnetic field) and LIU (low intensity ultrasound) applications. PC3-luc cells were seeded on a sterile 96-well plate (3×104 cells/well) and incubated overnight at 37° C. in a 5% CO2 incubator for cell adhesion. Then all the medium was removed from each well and the adherent cells were rinsed 2 times with DMEM without phenol red (white DMEM). 100 μl of (M-CA)f or (M-CMD)f suspension prepared at 1 mg/ml of iron in white DMEM supplemented with 10% v/v of FBS, 1% v/v of penicillin-streptomycin and 1% v/v of HEPES buffer were added to each well. Subsequently, all the wells were re-incubated for 3 h at 37° C. in a 5% CO2 incubator. After that, the hyperthermia experiment was divided into three conditions in which there were for each condition 3 wells containing only Pc3-luc cells, 3 other wells containing cells incubated with (M-CA)f, and 3 other wells of cells with (M-CMD)f. In the first condition, the wells were not exposed to any hyperthermia source. in the second condition, 3 wells per time were placed in a polystyrene holder positioned at the center of a copper coil, in which an AMF of 42 mt and 195 KHz was applied for 30 min with an easy heat equipment to heat the cells. in the third condition, each of the 3 wells was soaked in an adaptor filled with degassed water to make the connection with an ultrasound plane transducer (3.6 cm in diameter) and thus to be exposed for 30 min to ultrasound intensity lying between 0.3 and 1 w/cm2, at 1 MHz and in continuous mode, produced by ultrasound generator (Primo Therasonic 460). To limit the evaporation of culture medium, each well was covered by a cap when exposed to a hyperthermia source. The temperature was monitored over time using a flexible thermocouple probe (physitemp, it-18) inserted into each well and Dasylab software. the SAR of the sample was calculated using the following equation: SAR (W/gFe)=(Cmedium/CFe)×(Δt/δt)
where, Cmedium=4.2 j·g−1·k−1 is the specific heat capacity of water, Cfe, which is measured in g of iron per g of water, represents the iron concentration in the magnetosome sample, and Δt/δt (measured in ° C./s) is the slope of temperature variation with time of the magnetosome sample after subtracting that of the control samples. Following the hyperthermia treatments, all the wells were incubated for 24 h at 37° C. in a 5% CO2 incubator. After that, the cell viability in each well was determined using resazurin reagent.
Quantification in the iron of magnetosomes internalized by cells. After taking all the supernatant for the cell viability assay, PC3-luc cells adhered at the bottom of the wells were delicately rinsed 3 times with 300 μl of 1×pbs. Each well was then incubated with 50 μl of 0.25% trypsin—EDTA for 1 min at 37° C. and 5% CO2, followed by the introduction of 150 μl of complete DMEM. After that, all the liquid in each well (200 μl) was transferred into a 1.5 ml-eppendorf tube. After a few seconds of vortexing, a sample of 20 μl was taken from the Eppendorf tube and mixed with 20 μl of trypan blue to determine cell concentration using a Malassez hemocytometer. 180 μl of the remaining cell suspension were centrifuged at 14100 g for 20 min to remove all the supernatant. The cell pellet was next dissolved overnight in 20 μl of 37% w/v HCl and 145 μl of 70% w/v HNO3 at ambient temperature. Finally, the solution was diluted with ultrapure water in a total volume of 5 ml and analyzed in an inductively coupled plasma-mass spectrometer (agilent, 7900 icp-ms) to determine the magnetosome iron content internalized into PC3-luc cells.
Statistical analyzes. The data were analyzed using graphpad prism version 8.0. All measurements were performed in triplicate (n=3) and data were reported as the mean±standard deviation (sd). Statistical comparisons were carried out using one-way anova and the differences were considered significant when * p<0.05, ** p<0.01, and *** p<0.001.
Magnetosome formulation: summary of the previously published first steps and presentation of the additional/complementary steps proposed in this study. This study presents additional aspects concerning the implementation of a magnetosome-based formulation for injection in humans. The first steps of this formulation, are summarized. step 1 consists in cultivating magnetotactic bacteria, leading to magnetotactic bacteria of spirilla shape containing chains of magnetosomes in their cytoplasm with magnetosome mean sizes of 36.6 (standard deviation of 6.4 nm). Step 2 involves magnetosome extraction from magnetotactic bacteria issued from step 1, using KOH bacterial lysis and magnetic separation of magnetosomes from bacterial debris, leading to extracted magnetosomes arranged in chains (Mg—Ch), where each magnetosome is surrounded by a bacterial inflammatory membrane, which seems too immunogenic in the absence of specific treatment for injection to human. Step 3 comprises the purification of Mg—Ch through combustion to remove/denature bacterial organic material from Mg—Ch, leading to non-pyrogenic magnetosome minerals (M-uncoated) with a low percentage of carbon of 0.2%, which tend to aggregate due to their bare surface. M-uncoated are coated with two biocompatible compounds, i.e. citric acid (CA) or carboxy-methyl dextran (CMD), yielding non-pyrogenic coated magnetosome minerals (M-CA and M-CMD) reconstituted in chains.
The objective of this example is to add some further steps in the preparation of coated magnetosomes to be able to store M-CA and M-CMD over a long period of time, while preventing coating degradation. To achieve this aim, M-CA and M-CMD have been kept in the form of an anhydrous powder, which can be reconstituted in a NP suspension at any time, depending on need. We have then verified that reconstituted M-CA and M-CMD, designated as (M-CA)f, (M-CMD)f, (M-CA)fw, (M-CMD)fw for lyophilized M-CA and M-CMD before and after NP resuspension in water, respectively, have preserved their physicochemical properties and anti-tumor activity.
Formulation of M-CA and M-CMD in the presence of cryo-protectant for long term storage. Here, we determine the conditions of fabrication of an injectable magnetosome formulation with a long-term stability. The latter is evaluated by measuring the percentage of absorption stability, measured at 480 nm over time, of a suspension of magnetosomes, preferentially of 50 mg of magnetosomes during 2 hours following its homogenization. The percentages of 100% and 0% correspond to stable and unstable suspensions, respectively. we chose to carry out stability measurements with 50 mg of magnetosomes, since we anticipate to administer this amount of magnetosomes in a human prostate tumor of 1-2 cm diameter based on our pre-clinical mouse efficacy data extrapolated to humans. furthermore, a lapse of time of 2 hours was selected for stability assessment, which seems to be long enough for a nurse or doctor to be able to administer a stable magnetosome suspension in tumors. Given that the purified magnetosome minerals devoid of active bacterial organic matter (M-uncoated) were unstable, i.e. their percentage of stability is 0%, they were coated with CA and CMD, leading to M-CA and M-CMD, which are perfectly stable, i.e. their percentage of stability is 100%, which is larger to that of M-uncoated and M-gC. When M-CA and M-CMD are lyophilized in the absence of a specific compound that can preserve their coating, they form NP powders designated as (M-CA)fd and (M-CMD)fd characterized by the presence of agglomerated magnetosomes. When (M-CA)fd and (M-CMD)fd are resuspended in water following the lyophilization step, they rapidly sediment leading to a percentage of stability lower than 5%. To enable the reconstitution of M-CA and M-CMD in stable NP water suspension following lyophilization, i.e. without damaging M-CA/M-CMD coating, M-CA and M-CMD at the foreseen therapeutic dose of 50 mg/ml were mixed with various cryo-protectants, i.e. glucose, mannitol, sorbitol, sucrose, or trehalose, used at percentages in mass of 5% and 10%. after being lyophilized, such formulations were resuspended in water to determine their colloidal stability. Considering M-CMD first, only 5% and 10% sorbitol succeed in maintaining 100% colloidal NP stability. Furthermore, the osmolality of the M-CMD suspension increases with the percentage of sorbitol added to the M-CMD suspension, from 0 mosm/kg H2O in the absence of cryoprotectant to 280 and 560 mosm/kg H2O for 5% and 10% of sorbitol, respectively. M-CMD formulated with 5% sorbitol leads to an osmolality value lying within the range of plasma osmolality (275-290 mosm/kg H2O), which is acceptable for human injection. Turning now to M-CA, M-ca mixed with 10% of glucose, sorbitol or sucrose, leads to 100% colloidal stability. However, such formulations are hypertonic, i.e. their osmolalility is larger than 300 mosm/kg H2O. To solve this issue, we selected M-CA mixed with 10% sucrose that leads to an osmolality of 330 mosm/kg H2O, which is the closest value to the plasma osmolality, and we added to such mixture either PEG 4000 or Dextran T1, to decrease the osmolality of the formulation to the plasma osmolality. Whereas the use of Dextran t1 enables stabilizing the formulation, it yields an osmolality that is lower than the plasma osmolality. Hence, Dextran T1 was abandoned. By contrast, by combining sucrose with PEG 4000, a 100% stable formulation was reached while the sole addition of 7.5% sucrose or 15% PEG 4000 led to a stability at about 90 or 50%, respectively. This result suggests a synergistic effect between sucrose and PEG 4000, which appear to be mutually supportive, i.e. PEG 4000 could effectively protect NP during freezing by encapsulating NP in a glassy matrix, and amorphous sucrose could serve as a “water replacement” reservoir allowing the formation of hydrogen bonds with citric acid on NP surface, and consequently preserving the coating integrity against the stress induced by dehydration. The optimal combination of sucrose and PEG 4000 corresponds to 1.25% of PEG 4000, i.e. the lowest concentration of PEG 4000 to prevent the risk of molecular mobility and degradation of the formulation during storage, which could be a consequence of a too large PEG concentration, and the largest percentage in mass of sucrose, i.e. 3.75%, to yield stable formulated M-CA in suspension. The M-CA formulation with 1.25% of PEG 400 and 3.75% of sucrose displays a osmolality of 225 mOsm/Kg H2O, which can be adjusted to be isotonic upon reconstitution.
M-CA/M-CMD preserve their physicochemical properties following their formulation. To examine if magnetosomes formulated with 5% sorbitol for M-CMD, or 1.25% of PEG 400 and 3.75% of sucrose for M-CA, which are designated as (M-CMD)f and (M-CA)f before removal of cryoprotectant and as (M-CMD)wf and (M-CA)wf after removal of cryoprotectant, have maintained their physicochemical properties, we have compared the percentages of carbon and nitrogen as well as the FT-IR spectra and surface charges of formulated magnetosomes with those of magnetosomes harvested at the various steps preceding formulation, i.e. M-CA, M-CMD, M-gC, and M-uncoated. Concerning the percentages in mass of carbon and nitrogen in magnetosomes at the various formulation steps, they first decrease from carbon and nitrogen percentages of % C=22.7% and % N=2.2% in M-gC down to % C=012% and % N=0.01% in M-uncoated, corresponding to the removal of the magnetosome membrane in M-gC to yield M-uncoated. Then, they increase a first time from M-uncoated to M-CA (% C=1.61%, % N=0.01%) and M-CMD (% C=3.80%, % N=0.01%), which is due to the addition of a coating of CA and CMD to the surface of M-uncoated, and a second time from M-CA and M-CMD to (M-CA)f (% C=18.60% and % N=0.01%) and (M-CMD)f (% C=18.56% and % N=0.01%), as the cryoprotectant is added to the formulated magnetosomes. In the final step, these percentages decrease again following the washing of the cryoprotectant from the formulated magnetosomes, i.e. down to % C=1.6% and % N=0.01% in (M-CA)fw and % C=3.83%, % N=0.01% in (M-CMD)fw. The FT-IR spectra of the magnetosomes at the various steps of their formulation further confirm the CHNS trends described above. Indeed, Mg-C FT-IR spectrum displays vibration bands of P—O (at 1037 cm−1), C—O (at 1410 cm−1), N—H (at 1542 and 1641 cm−1), and O—H (at 3276 cm−1), corresponding to functional groups present in the phospholipid magnetosome membrane. Concerning M-uncoated, its FT-IR spectrum does not indicate the presence of peaks above 1100 cm−1, which is consistent with the removal of most organic material in this sample. The two peaks at 612 and 693 cm−1 are attributed to Fe-0 stretching vibrations, which come from the iron oxide comprised in the magnetosome mineral core, and are therefore present in all FT-IR spectra of the different types of magnetosomes. The coating of M-uncoated with CA and CMD leads to a series of peaks in the range of 1000-1300 cm1 and 1630 −1750 cm−1, which correspond to the stretching vibrations of c-o and c=o, respectively, attributed to the carboxylate groups of CA and CMD in M-CA and M-CMD. Further on in the formulation process, lyophilized magnetosomes (M-CA)f and (M-CMD)f display intense bands of alcohol and alkane groups, e.g. the stretching vibration bands of C—O, O—H and C—H in the range of 970-1250 cm−1, 3200-3550 cm−1 and 2850-3000 cm−1, respectively, corresponding to sucrose and PEG 4000 in (M-CA)f, and sorbitol in (M-CMD)f. Finally, following a washing step, (M-CA)fw and (M-CMD)fw display FT-IR spectra, which are very close to those M-CA and M-CMD, indicating that the washing procedure has easily separated the cryoprotectant from the coated magnetosome, due to the absence of a strong binding between cryoprotectant molecules and magnetosome coating agents. CHNS and FT-IR measurements suggest that the surface of formulated magnetosomes remains unchanged compared with that of coated magnetosomes before formulation. To confirm this deduction, we have measured the surface charges of the formulated magnetosomes that we have compared with those of non-formulated magnetosomes. (M-CA)f and (M-CMD)f display very similar surface charges as those of M-CA and M-CMD when the pH of the suspensions containing these NP is varied between 2 and 10. Such behavior indicates that the cryoprotectant efficiently maintains magnetosome surface charges in (M-CA)f and (M-CMD)f, possibly by efficiently protecting the coating layers in M-CA and M-CMD against degradation/removal during freeze-drying, and by preventing the cryoprotectant to be strongly bound to NP. Furthermore, under conditions of use for human injection, i.e. at the therapeutic dose of 50 mg/ml in iron and at pH of −6.5, (M-CA)f and (M-CMD)f display surface charges that are below −30 my, creating the conditions of strong repulsive electrostatic forces in (M-CA)f/(M-CMD)f suspensions, which can prevent NP agglomeration caused by magnetic dipole interactions and stabilize these NP in suspension. Electron microscopy measurements conducted on formulated magnetosomes washed of their cryoprotectant, i.e. on (M-CA)fw and (M-CMD)fw, show that formulated magnetosomes preserve a chain arrangement after the freeze-drying step, a property complementary to that of their surface charge, ensuring their stability and preventing their aggregation. Finally, the long-term stability of formulated magnetosomes, which is sought for here, is ensured when the amount of water remaining in the freeze-dried product after the freeze-drying process, called residual moisture (RM), is sufficiently small, i.e. typically below 3% for a pharmaceutical product. The RM content of (M-CA)f and (M-CMD)f were measured by introducing these NP in a TGA, by heating them at 120° C. during 1 h with a heating rate of 6° C./min, and by measuring the weight loss of these NP attributed to water evaporation. The RM contents of (M-CA)f and (M-CMD)f lie within the rage of (1.8-2.4)%, which are pharmaceutically acceptable. Furthermore, due to this efficient lyophilization process, (M-CA)f and (M-CMD)f could be resuspended in water, preferentially after 6 months, where they maintain their stability after a storage time of half a year. Sterility/non-pyrogenicity of formulated magnetosomes. Compared to unformulated magnetosomes, formulated magnetosomes must not only preserve their physicochemical properties, but also their biocompatibility. To study this last aspect, we first examined the sterility of the formulated magnetosomes by introducing (M-CA)f and (M-CMD)f during 14 days in solutions of tryptic soy broth (TSB) and fluid thioglycolate medium (FTM) at 30° C. and 37° C., respectively. indeed, these conditions are known to amplify bacterial contaminants when they are initially present in a tested sample, and therefore to enable their detection. To determine the presence (or absence) of bacteria in (M-CA)f and (M-CMD)f, the optical density of these NP in suspension was measured at 600 nm (OD600) at the end of the incubation time. The value of OD600 reflects the turbidity of these suspensions, which is associated with the presence of potential bacterial contaminant (M-CA)f and (M-CMD)f display a low value of OD600<0.1, which is comparable to OD600<0.1 of M-uncoated and of the sterile TSB and FTM media before incubation (NC), indicating the absence of bacterial contaminants in (M-CA)f and (M-CMD)f. This result is further confirms the absence of colonies in agar plates inoculated with (M-CA)f and (M-CMD)f. To complement the assessment of the sterility and non-pyrogenicity of (M-CA)f and (M-CMD)f, the endotoxin concentration of these NP was measured. (M-CA)f and (M-CMD)f appear to contain a very low endotoxin concentration of 2-5 EU/mg of iron, comparable to the values measured for M-uncoated. Taken altogether, these results highlight the efficacy of the magnetosome purification step in removing bacterial contaminants from M-gC, whose initial presence before a specific treatment is revealed by the high M-gC OD600 value of 1-2.5 as well as a large M-gC endotoxin concentration of 50 EU/mg in iron. Such behavior can be attributed on the one hand to a double step de-pyrogenation process, i.e. first by mixing M-gC with KOH at 80° C. and second by heating M-gC above 400° C., and on the second hand to formulation steps carried out under ascetic conditions, i.e. the coating of magnetosome minerals with CA or CMD, the addition of cryoprotectant to coated magnetosomes M-CA and M-CMD, the lyophilization of the obtained mixture, are carried out under sterile hood.
Non-cytotoxicity of formulated magnetosomes. To examine the cytotoxicity of the lyophilized formulated magnetosomes, 100 μl of (M-CA)f and (M-CMD)f suspensions of concentrations varied between 0.001 and 1 mg of NP in iron per ml were brought into contact with various mammalian cell lines, i.e. 3T3, L929, and V-79 cells, during 24 hours at 37° C. Following this treatment, the cell viability was assessed by measuring cellular metabolic activity with the resazurin assay. All cell lines display a viability larger than 70% for all tested concentration of (M-CA)fw and (M-CMD)fw, revealing that these NP are non-cytotoxic under the tested conditions, and that the presence of a cryoprotectant and of a lyophilization step in the magnetosome formulation does not result in added cytotoxicity. Efficient hyperthermia treatment using formulated magnetosome minerals excited under alternating magnetic field and ultrasonic sources. We study if (M-CA)f and (M-CMD)f maintain the therapeutic activity observed in non-fully formulated magnetosomes M-CA and M-CMD. More specifically, we examine if (M-CA)f and (M-CMD)f efficiently destroy prostate tumor cells when 100 μl of (M-CA)f or (M-CMD)f at a concentration of 1 mg of np in iron per nil of water suspension are incubated with PC3-luc cells during 3 h, and the resulting mixture is exposed during 30 minutes to an AMF of 42 mt and 195 KHz or to a low intensity ultrasound of 0.3-1 w/cm2 and 1 MHz. Such conditions of excitation lead to a temperature increase of these mixtures from ambient temperature of 22-25° C. to moderate hyperthermia temperature of 46° C., where the temperature rise is significantly faster with the ultrasonic than with the magnetic excitation, leading to the temperature being maintained at 46° C. for almost 30 minutes and slightly less than 10 minutes with the ultrasound and magnetic field, respectively. The specific absorption rates (SAR) of (M-CA)f and (M-CMD)f are measured under exposure of these NP to AMF/LIU after subtraction of the initial temperature increase due to AMF/LIU in the absence of NP. While (M-CA)f and (M-CMD)f display a large SAR value of 229±16 W/gF, under AMF excitation, they produce a close to zero SAR value under LIU, highlighting the different nature of the heating with the two excitation sources, i.e. a heating originating from NP excitation by the AMF or absorption of ultrasound energy by cells, using AMF or LIU application, respectively. Moreover, the heating properties of the two types of formulated magnetosomes, i.e. (M-CA)f and (M-CMD)f are similar. This indicates that the differences between the two coatings in nature/composition or at least one constituent of the composition, coating thicknesses, carbon contents, do not significantly affect (M-CA)f and (M-CMD)f heating properties. We may conclude that certain heating contributions such as the Brownian one in the induction mechanism, which can depend on the coating properties, are not dominantly involved in the observed heating properties. To further assess the efficacy of the heat treatment in destroying tumor cells, Pc3-luc cells treated as described above were re-incubated overnight at 37° C. under 5% CO2. Their viability was subsequently measured. In the absence of a heat treatment, PC3-Luc cells incubated with (M-CA)f and (M-CMD)f display a viability of 80%, indicating the absence of cytotoxicity of (M-CA)f and (M-CMD)f towards these cells. This behavior agrees with that observed with healthy mammalian cells. By contrast, in the presence of M-CA/M-CMD and a heating session, the cell viability decreases by 30-40% and 60-85% following AMF and LIU application, respectively, compared with the two controlled conditions in which the cells are either only incubated with (M-CA)f/(M-CMD)f without heating or only exposed to AMF/LIU without NP exposure. These results suggest that LIU leads to more efficient cellular destruction than AMF, possibly due to a temperature maintained at 46° C. for a longer period of time with LIU than with AMF. Furthermore, it seems that although both types of formulated magnetosomes form an efficient pair with LIU to destroy prostate tumor cells, M-CMD outperforms M-CA, a behavior that we are currently trying to understand. Given that NP internalization in tumor cells is often correlated with efficient cellular destruction, we estimated the amount of (M-CA)f and (M-CMD)f, which is internalized in PC3-luc cells following a similar treatment as that used to measure cell viability. Following treatments, we have destroyed and dissolved the tumor cells, and measured their iron content by ICP-MS. The quantities of NP internalized in PC3-luc tumor cells, is larger for (M-CMD)f than for (M-CA)f in all tested conditions, i.e. with/without AMF/LIU treatments. Qi are estimated as 17-20 and 30 pg of NP per cell following AMF/LIU treatment for (M-CA)f and (M-CMD)f, respectively. Compared to (M-CMD)f, (M-CA)f displays a lower internalization in PC3-luc cells by a factor of 1.5-2. This behavior may be attributed to the more negative surface charge at physiological pH for (M-CMD)f than for (M-CA)f, possibly resulting in a better affinity of (M-CMD)f for the cationic sites of the plasma cellular membrane and subsequently to the higher cellular internalization possibly via pinocytosis for (M-CMD)f than for (M-CA)f. Considering now the potential impact of NP internalization on cell viability, it can be observed that the increase in NP internalization between M-CMD and M-CA is either correlated with an enhanced cellular mortality following LIU treatment or uncorrelated with a change in cellular mortality under AMF application. Such behaviors could be explained by ultrasounds acting synergistically with internalized NP to destroy tumor cells, i.e. ultrasounds could destroy/inactivate the cell membrane while NP could act perturbatively inside the cells from their intracellular location. By contrast, whereas AMF is expected to produce local heating at NP site, it is not supposed to act perturbatively against the cellular membrane. Thus, it is possible that in the presence of internalized NP, ultrasounds act more perturbatively than AMF, resulting in more efficient cellular destruction.
This example describes the experimental protocol used to dope magnetosome with zinc by growing MSR-1 magnetotactic bacteria (MTB) in 2-liter bottles in presence of a source of zinc, resulting in the production of zinc-doped magnetosomes.
The composition or at least one constituent of the compositions of the pre-growth medium and growth medium are indicated for 1 liter of medium in tables 1, 2 and 3 for the tested conditions. As a whole, 8 different culture conditions Zn0, Zn0.1, Zn1, Zn5, Zn10, Zn20, Zn50 and Zn100 are tested corresponding to 8 different concentrations of zinc added to the growth medium 0, 0.1, 1, 5, 10, 20, 50, 100 micromoles respectively. During the first day of the experiment (D1), a first step consists in collecting the tubes of 2 ml containing 1 mL of MSR-1 cellular stock tube from the freezer at −80° C., in thawing the tubes by letting them at room temperature for 10 minutes. Under a hood, we collected from these tubes 1 ml comprising 5.108 MSR-1 magnetotactic bacteria that we have inserted in 500 mL bottles filled with 250 mL of pre-growth medium. The 500 nil bottles are incubated for 7 days between D1 and D7 in an incubator at 29.5° C. and between D6 and D7 under orbital agitation at 110 rpm. During the seventh day (D7), the growth step started. For that, 2-liter bottles are filled with 1 L of deionized water and autoclaved. The 2-liter bottles are placed under the hood and then filled with 250 mL of filtered growth medium containing an iron source and increasing concentrations of a source of zinc to enable the synthesis of zinc doped-magnetosomes by MSR-1 MTB. The 500 mL bottles from the pre-growth step, are placed under a hood. 40 mL of the pre-growth medium containing the MSR-1 bacteria are transferred in the 2-liter bottles. The 2-liter bottles are incubated for 7 days between D7 and D14 in an incubator at 29.5° C. under orbital agitation at 110 rpm enabling the growth of MSR-1 magnetotactic bacteria and the synthesis of magnetosomes by these bacteria. A positive magnetic response (≥80%) at D14 and a ratio between the optical density at D14 and the optical density at D7 larger than 8.5 is observed for the conditions Zn0, Zn0.1, Zn5, Zn10 and Zn20. In contrast, very low magnetic response is observed for the conditions Zn50 and Zn100, with the highest tested concentrations of zinc of 50 and 100 micromoles respectively. In conclusion the conditions that enable the growth of MSR-1 bacteria with a large increase in optical density (OD565nmD14/OD565nmD7 larger than 8.5) and the synthesis of magnetosomes (positive magnetic response ≥80%) are Zn0, Zn0.1, Zn 1, Zn5, Zn10 and Zn20 with zinc concentrations of 0, 0.1, 1, 5, 10, 20 micromoles respectively as presented in table 4.
Following fermentation at D14, MSR-1 MTB from the 2-liter bottles (conditions Zn0, Zn0.1, Zn1, Zn5, Zn10, Zn20, Zn50 and Zn100) are concentrated by centrifugation at 4000 rpm during 30 min and stored at −20° C. for no less than 24 hours. Concentrated MSR-1 MTB are thawed at 70° C. for 1 h, resuspended with 30 milliliters of deionized water and transferred in 20 mL glass bottles. MSR-1 MTB are then lysed in 2 M KOH at 80° C. for 2 hours, in a sonicating bath at 25 Kilohertz. A direct current field is then applied to the suspension for 12 hours to separate magnetosomes from organic material residues. Magnetosomes are washed 2 times with 1 X, Phosphate-buffered saline (PBS) then 3 times with deionized water. Magnetosomes are collected in 50 ml-conical tubes and centrifuged at 10° C., 3380 g for 30 min. Tubes containing magnetosomes are stored at −80° C. for 24 hours and lyophilized at −50° C., 0.003 mbar for 24 hours. Extracted zinc doped-magnetosomes are thus obtained for the tested conditions Zn0, Zn0.1, Zn1, Zn5, Zn10, Zn20, Zn50 and Zn100.
The content of zinc in extracted zinc doped-magnetosomes is determined by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-AES). One milligram of extracted zinc doped-magnetosomes from conditions Zn0, Zn0.1, Zn1, Zn5, Zn10, Zn20, Zn50 and Zn100 are inserted in 15 mL tubes filled with 200 microliters of hydrochloric acid 12 Mole. The tubes of 15 milliliters containing the magnetosomes are vortexed and incubated at room temperature for 2 hours and then filled with 9.8 mL of nitric acid 2%. Analysis is then carried out by ICP-AES. The Zn/Fe ratio in extracted zinc doped-magnetosomes from the conditions Zn0, Zn0.1, Zn1, Zn5, Zn10, Zn20, Zn50 and Zn100 are presented in FIG. 2. For the condition Zn0 and Zn0.1, when no source of zinc or very low concentration of it is brought to the growth medium the percentage of zinc with respect to the iron in magnetosomes is very low at 0.14% and 0.16% respectively. However, we observed increasing percentage of zinc with respect to the iron in magnetosomes for the conditions Zn1, Zn5, Zn10, Zn20 and Zn50 at 0.26, 0.84, 1.36, 2.4 and 5.8% respectively. Finally for the condition Zn100 no zinc is detected in magnetosomes.
This example describes the experimental protocol used to dope magnetosome with zinc by growing MSR-1 MTB in presence of a source of zinc, resulting in the production of zinc-doped magnetosomes.
In this example, MTB are grown in 6.5-liter Applikon fermenters with an ez control system and piloted by the BioXpert software for the regulation of stirring and airflow rates. The composition or at least one constituent of the compositions of the pre-growth medium, growth medium, and fed-batch medium are indicated for 1 liter of medium in tables 1, 2 and 5 for the tested conditions. As a whole, 2 different culture conditions are tested: Condition A, in absence of a source of zinc and Condition B in presence of a source of zinc in the fed-batch medium at a concentration of 452 micromoles. During the first day of the experiment (Dl), a first step consists in collecting the tubes of 2 ml containing 1 mL of MSR-1 cellular stock tube from the freezer at −80° C., in thawing the tubes by letting them at room temperature for 10 minutes. Under a hood, we collected from these tubes 1 nil comprising 5.108 MSR-1 magnetotactic bacteria that we have inserted in 500 mL bottles filled with 250 mL of pre-growth medium. The 500 nil bottles are incubated for 8 days between D1 and D8 in an incubator at 29.5° C. and between D7 and D8 under orbital agitation at 110 rpm. During the eighth day (D8), the second pre-growth step started. For that, 3-liter fermenters are filled with 1 L of deionized water and autoclaved. The fermenters are placed under the hood and then filled with 500 mL of filtered pre-growth medium. The 500 mL bottles from the first step of pre-growth, are placed under a hood. 500 mL of the pre-growth medium containing the MSR-1 bacteria were transferred in the 3-liter fermenters. The temperature was maintained at 29.5° C. and agitation at 110 rpm in the 3-liter fermenters during 48 hours between D8 and D10. During the tenth day (D10), the growth step started. For that, 6.5-liter fermenters are filled with 3-liter of deionized water and autoclaved. The fermenters were then filled with 500 mL of filtered growth medium. The 3-liter fermenters from the second step of pre-growth, are placed under a hood. About 900 mL of pre-growth medium containing the MSR-1 bacteria originating from the second step of pre-growth is then transferred respectively in the 6.5-liter fermenters. Between day D10 and D18, an acidic fed-batch medium comprising a source of iron (condition A) and sources of iron and zinc (condition B) were added to the growth medium to enable the synthesis of magnetosomes (condition A) and zinc-doped magnetosomes (condition B) by MSR-1 bacteria, while maintaining the pH of the growth medium at 6.9. During the growth step, the temperature was maintained at 29.5° C., the stirring fixed at 150 rpm (rotation per minute) and airflow rates were adjusted to maintain to oxygen concentration above 0.1% and below 1% enabling the growth of MSR-1 magnetotactic bacteria and the synthesis of magnetosomes by these bacteria. The optical densities, measured at 565 nm of the bacterial suspensions, the volume of Fed-batch added to the medium at different days of the pre-growth step (D8 and D10) and growth step (D10, D12, D16, D17, D18) are indicated in table 6.
Following fermentation at D18, MSR-1 MTB are washed and concentrated in deionized water using tangential flow filtration to an OD565˜30 and stored at −80° C. for no less than 24 hours. Concentrated MSR-1 MTB from conditions A and B are thawed at 40° C. for 2 h and diluted with deionized water to an OD565˜20. MSR-1 MTB are then lysed in 2 M KOH at 80° C. for 1 h under agitation (150 rpm). A direct current field is then applied to the suspension for 12 hours to separate magnetosomes from organic material residues. Magnetosomes are washed 2 times with 10× Phosphate-buffered saline (PBS) then 3 times with deionized water. Magnetosomes are collected in 50 nil-conical tubes and centrifuged at 10° C., 3380 g for 30 min. Tubes containing magnetosomes are stored at −80° C. for 24 hours and lyophilized at −50° C., 0.003 mbar for 24 hours. Magnetosomes are then purified in a muffle furnace as followed, 50° C. (1 h)-270° C. (2 h)-310° C. (2 h)-420° C. (2 h). Purified magnetosomes (condition A) and purified Zinc-doped magnetosomes (condition B) are thus obtained.
The content of zinc in purified Magnetosomes (condition A) and in purified zinc-doped magnetosomes (condition B) is determined by inductively coupled plasma-mass spectrometer (ICP-MS). Two milligrams of purified Magnetosomes are dissolved in 35-microliter of 37% w/v HCl and 286-microliter of 70% w/v HNO3 at ˜20° C. for 12 hours. Ultrapure water is added to a volume of 10-milliliter. Analysis is then carried out by ICP-MS. The Zn/Fe ratio in purified magnetosomes (condition A) and purified zinc-doped magnetosomes (condition B) are presented in FIG. 2. When no source of zinc is brought to the growth medium (condition A), the percentage of zinc with respect to the iron in purified magnetosomes is very low at 0.06% while with a concentration of zinc sulfate of 30 micromoles in the growth medium the percentage of zinc with respect to the iron in purified zinc-doped magnetosomes (condition B) is 0.74%.
This example describes the experimental protocol used to coat purified zinc-doped magnetosomes originating from MSR-1 MTB with citric acid and a derivative of Protoporphyrin IX.
In the dark and under an inert atmosphere, the corresponding photosensitizer Protoporphyrin IX (=P1, CAS no 553-12-8) was dissolved in 20-50 mL of chloroform (CAS no 67-66-3). N-hydroxysuccinimide (3 eq., CAS no 6066-82-6) and N,N′ dicyclohexylcarbodiimide (3 eq., CAS n° 538-75-0) are added to the photosensitizer solution. The mixture was stirred at 40° C., for 4-24 hours, under argon. Purification of the crude material is performed using a silica gel column with DCM (Dichloromethane, CAS no 75-09-2). P1-NHS is obtained, after recrystallization in DCM/hexane (CAS no 110-54-3).
P1-NH S (1 eq) and N-Boc-2,2′-(ethylenedioxy)diethylamine (1 eq., CAS no 153086-78-3) are dissolved in 30 mL of THF (Tetrahydrofuran, CAS no 109-99-9). The reaction mixture is stirred at room temperature, under a nitrogen atmosphere and in the dark for 18 hours. The solvent is removed under vacuum. Purification of the crude material is performed using a silica gel column with a solution of ethanol and chloroform and P1-NHBoc is obtained. It is dissolved in 2 mL of TFA (Trifluoroacetic acid, CAS no 76-05-1). The solution is stirred for 2 hours at room temperature, in the dark and under nitrogen. Next, TFA is removed by lyophilization. The residue is solubilized in 10 mL of DCM, and anhydrous potassium carbonate is added until the color changed. After filtration, the organic layer is concentrated. Purification of the crude material is performed using a silica gel column with a solution of DCM and MeOH at different gradients (v/v). P1-NH2 is thus obtained.
In the dark and under an inert atmosphere, folic acid or citric acid (1 eq.) and N, N′ dicyclohexylcarbodiimide (1 eq.) is dissolved in 5 mL of anhydrous DMSO (Dimethyl sulfoxide, CAS no 67-68-5) and 2 mL of pyridine (CAS no 110-86-1) and the mixture is stirred for 15 min at room temperature. Compound P1-NH2 (0.9 eq.) is then added, and the reaction mixture is further stirred for 24 h at room temperature. The solution is slowly poured into vigorously stirred cold diethyl ether (CAS) no. The precipitate obtained by centrifugation is collected and washed with diethyl ether. The powder is dried under a high vacuum. The residue is purified by HPLC on a C18 preparative column using an acetonitrile/water gradient. P1-folic or P1-citric is thus obtained and presented in FIG. 3.
The coating process is carried out in the dark and under sterile conditions, using a biosafety hood. A P1-folic or P1-citric solution was prepared at a concentration of 50 mg/ml in ultrapure water, at pH 6 and is filtered with a 0.22 μm polyether sulfone filter for sterilization. Different volumes of this solution are added to the 50 milliliter-glass bottles containing 80 mg of purified zinc doped-magnetosomes (corresponding to 40 mg of Fe) previously dispersed in 16 ml of ultrapure water. The suspension is sonicated for 5 min in a sonicating bath (SB) (Elma, TI-H 20) at 25 Kilohertz at room temperature. Following sonication, the pH of the suspension is adjusted at 6 using NaOH at 1 Mole and the suspension is then sonicated in the SB at 25 Kilohertz, 80° C. for 10-30 h for preparing P1-folic purified zinc doped-magnetosomes (ZnM-P1-folic) or P1-citric purified zinc doped-magnetosomes (ZnM-P1-citric). After the sonication, the ZnM-P1-folic and ZnM-P1-citric suspension are centrifuged (3380) at 10° C. (Eppendorf, Centrifuge 5810 R) for 45 min to remove excess of P1-citric or P1-folic in the supernatant. ZnM-P1-folic and ZnM-P1-citric are then re-suspended in 20 nil of water and stored in the dark at +4° C.
| Number | Date | Country | Kind |
|---|---|---|---|
| 22020560.3 | Nov 2022 | EP | regional |
| 22020561.1 | Nov 2022 | EP | regional |
