Patent Application
20220118164
The invention relates to the field of medical devices, more particularly to devices enabling the joint extraction, from an organism, of at least one metal cation and at least one target molecule. The use of these devices makes it possible, for example, to prevent and/or treat pathologies linked to deregulation of the homeostasis of metals and/or of target molecules in the body, for example neurological diseases and/or proteinopathies.
Maintaining the homeostasis of an organism's internal environment, meaning of all the biological fluids or liquids of the organism, is necessary to the proper functioning of the organism. Systemic or local dysregulation of the homeostasis of metals and/or peptides or proteins has been demonstrated in many diseases.
For metals, chelation therapies aimed at reducing the concentration of metal ions have already been in use for many years in cases of acute poisoning. A number of chelating agents are thus already accepted for use in humans, each associated with a particular group of metals (G. Crisponi et al., Coordination Chemistry Reviews, 2015).
More and more scientific studies highlight the important role that metals could have in a number of neurological impairments, in particular iron, but also copper, zinc, manganese, and even aluminum and lead (E. J. McAllum et al., J. Mol. Neurosci., 2016). This is particularly the case for neurodegeneration with iron overload which is a rare disease associated with a genetic anomaly linked to an accumulation of iron in certain areas of the brain and which so far has only been treated palliatively (S. Wiethoff et al., Handb. Clin. Neurol., 2017). In addition, many studies have shown that iron tends to accumulate in the brain with age (J. Acosta-Cabronero et al., Journal of Neuroscience, 2016). Wilson's disease is also a genetic disease, causing an accumulation of copper in the body and leading to various problems, particularly hepatic and/or neurological (Anna Cztonkowskal et al., Nature Rev., 2018).
Several neurological diseases such as Alzheimer's, Parkinson's, and Huntington's disease are also accompanied by an increase in the amount of iron in specific areas, leading to cellular damage as well as oxidative stress (A. A. Belaidi et al., Journal of Neurochemistry, 2016). For example, Huntington's disease is a neurodegenerative disease resulting in movement disorders, cognitive decline, and psychiatric problems. In this disease, many markers of oxidative stress are observed in the brain, which may be linked to a deregulation of iron homeostasis (S. J. A. van den Bogaard et al., International Review of Neurobiology, 2013). The increase in the level of iron in several regions of the brain (putamen, caudate nucleus, and pallidum) has thus been validated by several MRI studies including that of Bartzorkis (G. Bartzorkis et al., Archives of Neurology, 1999).
In these same pathologies, the homeostasis of other biological compounds is also disrupted. In Alzheimer's disease for example, the A-β (amyloid-beta) peptide, a peptide of about 42 amino acids (39 to 43), accumulates to form amyloid-beta aggregates. Treatments for amyloid diseases by extracting the Aβ peptide from biological fluids have thus been proposed (US2013/0045216 A1; M. Menendez-Gonzalez et al., Hypothesis and Theory, 2018). Still with the aim of treating or slowing the development of Alzheimer's disease, it has also been proposed to dilute the cerebrospinal fluid by replacing and filtering this fluid, in order to decrease the levels of Aβ peptide and of abnormally phosphorylated Tau protein (phospho-Tau) (M. M. Gonzalez, Cureus, 2017).
In addition to Alzheimer's disease, in many other amyloid pathologies such as Parkinson's disease and prion disease, a conformational conversion of normal soluble proteins into insoluble proteins has been demonstrated, leading to the formation of amyloid fibrils or plaques. Antibodies or small molecules specifically targeting these proteins are thus being studied with the aim being to inhibit the key stages of the aggregation process of these abnormal proteins, reduce the conversion of proteins into their pathological conformation, reduce the toxicity of pathological proteins, or increase the selective clearance of abnormal proteins (N. Cremades et al, Neurobiol Dis., 2018).
In addition, in Alzheimer's disease in particular, interactions have been demonstrated between Aβ peptides and certain metal ions, in particular ions from metals such as zinc, iron, or copper, which can lead to increased protein aggregation (Tougu et al. Metallomics, 2010).
It is thus accepted that in many proteinopathies, metal cations play an important role in the formation of abnormal configurations of certain proteins: in particular, some promote the formation of aggregates, fibrils, or other solid deposits. In proteinopathies, there would therefore locally be a two-fold deregulation of homeostasis: deregulation of the homeostasis of certain metals and deregulation of the homeostasis of protein-type target molecules, causing aggregates and other solid deposits.
Although scientific knowledge relating to these various pathologies is advancing (Pfaender S. et al., 2014; Boland B. et al., 2018; Iadanza M. G. et al., 2018), to date there is no effective treatment for Alzheimer's disease or Parkinson's disease, and more generally for neurodegenerative diseases and more inclusively for diseases involving multiple deregulations causing dyshomeostasis.
There is therefore currently a need to develop new means enabling the prevention and/or treatment of pathologies involving multiple deregulations causing dyshomeostasis and the formation of aberrant protein conformations leading to the formation of deposits, aggregates, fibrils, or plaques comprising said proteins. These means would thus have one or more of the following advantages:
These advantages and many others are described in the present disclosure.
A device is proposed for the joint extraction of at least one metal cation and at least one target molecule from a biological fluid, a biological aggregate, an organ, or tissue, for diagnostic or therapeutic purposes, characterized in that it comprises:
Also proposed is a microdialysis system comprising said device for extraction and such that said system comprises at least:
According to an alternative embodiment, a dialysis system is provided comprising said device for extraction and such that it comprises at least:
The features set forth in the following paragraphs may optionally be implemented. They may be implemented independently of one another or in combination with one another.
The means for extraction of the metal cation may be a perfusion fluid used in a dialysis or microdialysis system, said perfusion fluid further comprising said ligand exhibiting specific affinity for the target molecule.
Advantageously, the complexation constant log(KC1) of the chelating agent for at least one metal cation is greater than 10, preferably greater than or equal to 15, and said at least one cation is selected from the cations of metals Cu, Fe, Zn, Hg, Cd, Pb, Mn, Co, Gd, and Al, alone or in combination, and more particularly Cu, Fe, and Zn, alone or in combination.
According to a preferred embodiment, the means for extraction makes it possible to extract the cations from a biological fluid, biological aggregate, organ, or tissue, when the content of said metal cations is less than 1 ppm, preferably less than 0.1 ppm, more preferably less than 0.01 ppm, and even more preferably less than 1 ppb.
The means for extraction may make it possible to extract a quantity of metal cations representing at least 1% of its mass, and preferably more than 10% of its mass.
The device is advantageously a perfusion fluid comprised in a dialysis system comprising a dialysis membrane and a reservoir comprising the perfusion fluid, said perfusion fluid being selected among:
Preferably, the device comprises a dialysis system comprising a dialysis membrane and a reservoir comprising a perfusion fluid, said perfusion fluid being selected among:
According to an advantageous embodiment, the chelating agents are obtained by grafting, onto the nanoparticles or onto the polymer, one of the following complexing molecules or derivatives thereof: DOTA, DTPA, EDTA, EGTA, BAPTA, NOTA, DOTAGA, DFO, DOTAM, NOTAM, DOTP, NOTP, TETA, TETAM, TETP and DTPABA, or mixtures thereof.
Preferably, the nanoparticles are nanoparticles based on polysiloxane with an average diameter of between 3 and 50 nm, comprising the chelating agent obtained by grafting DOTA, DOTAGA, EDTA, or DTPA onto the nanoparticles.
According to one particular embodiment, the chelating agent contains at least one alkaline earth cation, preferably a cation of metals selected among Ca and Mg.
Advantageously, at least 10% of the chelating agents of said device are precomplexed with an alkaline earth cation; preferably 20%, more preferably 30%, and even more preferably more than 50% of the chelating agents of said device are precomplexed with an alkaline earth cation.
According to one embodiment, the nanoparticles or polymers, comprising the chelating agent obtained by grafting DOTA, DOTAGA, EDTA, or DTPA, have an average diameter greater than 20 kDa and less than 1 MDa.
Advantageously, the nanoparticles are based on polysiloxane or the polymers are based on chitosan or polyethylene glycol or polyvinyl alcohols.
According to one embodiment, the target molecule is selected among proteins, peptides, and glycoproteins. Advantageously, the target molecule is selected among amyloidogenic proteins and components of amyloid structures (in native monomeric form, or in the form of oligomers, or fibrils or aggregates or molecules responsible for their formation or their accumulation).
According to a preferred embodiment, the target molecule is selected among molecules involved in amyloidoses, tauopathies, or any pathology involving a deposit based on one or more proteins. Advantageously, the target molecule is selected among proteins and/or their precursors such as immunoglobulin light and heavy chains, serum amyloid A protein, transthyretin, apolipoprotein AI, AII, AVI, CII, or CIII, beta-2 microglobulin, gelsolin, lysozyme, fibrinogen, cystatin C, atrial natriuretic factor, calcitonin, amylin, insulin, prolactin, lactoferrin, cadherin, ABri, ADan, amyloid-beta peptide, prion protein, alpha-synuclein, tau protein, superoxide dismutase, huntingtin, neuroserpin, actin, ferritin, or mixtures thereof.
Preferably, the ligand is an antibody or an engineered protein ligand of the target molecule. Advantageously, the ligand is selected among:
According to a preferred embodiment, the device is used in the treatment of:
Other features, details and advantages of the invention will be apparent from reading the detailed description below, and from analyzing the accompanying drawings, in which:
For the most part, the drawings and the description below contain elements that are certain in nature. They can therefore serve not only to better understand the invention, but also contribute to its definition where appropriate.
The inventors have developed a medical device enabling the joint extraction of at least one metal cation and at least one target molecule, preferably at least two target molecules, from a fluid, a biological aggregate, an organ, or tissue, for diagnostic or therapeutic purposes.
The term “joint extraction of at least one metal cation and at least one target molecule” is understood to mean the simultaneous extraction of said metal cation and of said target molecule or the successive extraction of said metal cation and of said molecule target in any order whatsoever, the extraction of these two compounds being carried out with a short period of time between the two extractions, meaning preferably less than 24 hours, more preferably less than 12 hours, even more preferably less 1 hour.
The term “a metal cation” is understood to mean at least one metal cation. If several metal cations are involved, they may be metal cations of the same type or of different types.
The term “a target molecule” is understood to mean at least one target molecule. If several target molecules are involved, they may be target molecules of the same type or of different types.
The term “ligand” is understood to mean a molecule which binds, preferably reversibly, to the target molecule in a specific manner. Advantageously, the specific binding of ligand-target molecule is achieved by virtue of forces between molecules, such as ionic bonds, hydrogen bonds, hydrophobic interactions, and van der Waals forces. The ligand-target molecule interaction is thus reversible and more or less strong depending on the number and nature of the bonds formed. The strength of this interaction is defined by the affinity for the target molecule, which is linked to the dissociation constant.
The extraction of said biological compounds, meaning of said metal cation and of said target molecule, has the goal of maintaining homeostasis in said compounds, for therapeutic or diagnostic purposes. Maintaining homeostasis means regulating the content of said compounds within an organism, in particular with the aim being to extract said compounds in excess, which can be responsible for pathologies. Said compounds may be in excess within a biological fluid or within a biological aggregate. The extraction of one of the components may also have the aim of bringing the concentration of at least one of the biological compounds to below the solubility threshold of the biological aggregates linked to the pathology and thus to reduce their formation and/or lead to their dissolution.
The term “biological fluid” is understood to mean any fluid produced by the organism to which it relates. It may or may not be a circulating fluid. More particularly, it may be blood, lymph, bone marrow, chyle, any interstitial fluid, cerebrospinal fluid (CSF) or more specifically cerebrospinal fluid or spinal fluid, synovial fluid, peritoneal fluid.
The term “biological aggregate” is understood to mean any accumulation of target molecules and/or metals in the form of fibrils, matrix compounds, or plaques. As an example, they may be amyloid components, for example in the form of fibrils or plaques, accumulations of tau proteins, fatty plaques in particular such as atheromatous plaques, etc.
According to the invention, the term “organ” means all organs which can be brought into contact with the device of the invention or within which said device can be implanted or inserted. Preferably, the organ(s) are selected among the brain, the liver, the pancreas, the intestines, and the lungs.
According to the invention, the term “tissue” means all tissues which can be brought into contact with the device of the invention or within which said device can be implanted or inserted. Preferably, the tissue or tissues are selected among the peritoneum and tumor tissue (where appropriate from a tumor). For example, said device can be placed in contact, inserted, or implanted by endoscopy, in particular within a tumor.
“At least one” is understood to mean one or more of the compounds in question, of the same type or of a different type.
The term “dialysis” is also understood to mean specific dialyses such as, for example, microdialysis.
The extraction device comprises a ligand exhibiting specific affinity for the target molecule. Thus, said compound is able to bind specifically to said target molecule. It may be an antibody, a nanobody, a peptide, a protein, or any other ligand able to bind specifically to the target molecule.
The term “antibody” is understood to mean an immunoglobulin formed of 4 polypeptide chains, two heavy H and two light L, capable of specifically binding an antigen, also called a target molecule in the context of the invention.
The term “nanobody” is understood to mean an antibody element capable of specifically binding an antigen or target molecule in the context of the invention.
“Engineered protein ligand” (“scaffold protein” or “engineered protein”) is understood to mean a compound or protein fragments selected for their affinity towards specific target molecules. They are generally lighter than antibodies, often easier to produce as well, and chemically stable. Advantageously, the engineered protein ligands are less than 50 kDA, preferably less than 30 kDa, and more preferably less than 3 kDa. Such ligands have a good specific surface area. These engineered protein ligands may be selected among: ABD, Adhiron, Adnectin, Affibody, Affilin, Affimer, Affitin, Alphabody, Anticalin, Armadillo repeat proteins, Atrimer/tetranectin, Avimer/Maxibody, Centyrin, DARPin1.
The extraction device further comprises a means for extraction of at least one metal cation, said means being a perfusion fluid comprising at least one chelating agent, said perfusion fluid being contained in a dialysis system.
According to the invention, the term “chelating agent” means an organic group capable of complexing with at least one metal cation. The complexation reaction can be a transmetalation, meaning an exchange of two metal cations. In such a case, the chelating agent may be precomplexed with a first metal cation which will subsequently be exchanged with the target metal cation.
In an advantageous embodiment, at least 10% of the chelating agents of said device are precomplexed with an alkaline earth cation, preferably 20%, more preferably 30%, and even more preferably more than 50% of the chelating agents of said device are precomplexed with an alkaline earth cation.
According to a preferred embodiment, the complexation constant log(KC1) of said chelating agent for at least one of said metal cations is greater than 10, in particular 11, 12, 13, 14, 15, and is preferably greater than or equal to 15. When the chelating agent is precomplexed with a first metal cation, the complexation constant log(KCl′) for the first metal cation is less than the complexation constant log(KCl) of the target metal cation.
Advantageously, the chelating agent, with a constant at least greater than or equal to 10 and preferably greater than or equal to 15, complexes at least one of the cations of the metals Copper (Cu), Iron (Fe), Zinc (Zn), Mercury (Fig), Cadmium (Cd), Lead (Pb), Aluminum (Al), Manganese (Mn), Arsenic (As), Mercury (Hg), Cobalt (Co), Nickel (Ni), Vanadium (V), Tungsten (W), Zirconium (Zr), Titanium (Ti), Chromium (Cr), Silver (Ag), Bismuth (Bi), Tin (Sn), Scandium (Sc), Yttrium (Y), Lanthanum (La), Cerium (Ce), Praseodymium (Pr), Neodymium (Nd), Samarium (Sm), Europium (Eu), Gadolinium (Gd), Terbium (Tb), Dysprosium (Dy), Holmium (Ho), Erbium (Er), Thulium (Tm), Ytterbium (Yb), Lutecium (Lu), Actinium (Ac), Uranium (U), Plutonium (Pu), Americium (Am), alone or in combination. Even more advantageously, the chelating agent complexes at least one of the cations of the metals Copper, Iron, Zinc, Mercury, Cadmium, Lead, Aluminum, Manganese, and Gadolinium, in particular Manganese and Gadolinium. Even more advantageously, the chelating agent complexes at least one of the cations of the metals Copper, Iron, and Zinc, alone or in combination.
Advantageously, the specificity of the chelating agent for said metal cation to be extracted is high relative to the other cationic trace elements, in particular the difference between the complexation constants is preferably greater than 3; and, more particularly, the difference between the complexation constants with calcium and magnesium is preferably greater than 3 and even greater than 5.
According to a preferred embodiment, said device, or more specifically the means for extraction of at least one metal cation, also contains trace elements selected among calcium, magnesium, iron, copper, zinc, and manganese within the perfusion fluid or even directly on said chelating agent. In the latter case, this involves a transmetalation reaction and the cations are specifically chosen to allow such a reaction. Such an embodiment makes it possible, for example, to regulate the homeostasis of essential metals. The chelating agent contains at least one alkaline earth cation, preferably a cation of metals selected among Ca and Mg.
According to one embodiment of the invention, the means for extraction of the metal cation allows extracting said metal cation from a biological fluid, a biological aggregate, an organ, or tissue, when its content is less than 1 ppm, in particular 0.1 ppm, 0.01 ppm, and is preferably less than 1 ppb.
According to an advantageous embodiment, said means for extraction of the metal cation allows extracting a quantity of metal cations representing at least 1% of its mass, and preferably more than 10% of its mass.
In addition, the extraction device comprises at least one ligand exhibiting specific affinity for a target molecule. Said target molecule is preferably selected among proteins, peptides, and glycoproteins, more preferably it is selected among components of amyloid structures. According to an advantageous embodiment, the target molecule comprises a specific peptide sequence recognized by the ligand.
According to one embodiment of the invention, the target molecule is selected among proteins, peptides, and glycoproteins.
Advantageously, the target molecule is selected among amyloidogenic proteins and components of amyloid structures, in particular in native monomeric form or in the form of oligomers, fibrils, or biological aggregates. The target molecule may also be one or more molecules responsible for the formation or accumulation of said oligomers, fibrils, or biological aggregates.
According to a preferred embodiment, the target molecule is selected among molecules involved in amyloidoses, tauopathies, or any pathology presenting a deposit based on one or more proteins.
According to one embodiment compatible with the preceding embodiments, the target molecule is selected among proteins and/or their precursors such as immunoglobulin light and heavy chains, serum amyloid A protein, transthyretin, apolipoprotein AI, AII, AVI, CII, or CIII, beta-2 microglobulin, gelsolin, lysozyme, fibrinogen, cystatin C, atrial natriuretic factor, calcitonin, amylin, insulin, prolactin, lactoferrin, cadherin, ABri, ADan, amyloid-beta peptide, prion protein, alpha-synuclein, tau protein, superoxide dismutase, huntingtin, neuroserpin, actin, ferritin, or mixtures thereof.
Preferably, the ligand is an antibody or an engineered protein ligand of the target molecule. Advantageously, the ligand is selected among:
According to one embodiment, the extraction device comprises a perfusion fluid comprising at least one chelating agent, said perfusion fluid being contained in a dialysis or microdialysis system or any miniaturized dialysis device, in particular with a fixed exchange reservoir.
Advantageously and according to a preferred embodiment, the dialysis or microdialysis system comprises:
According to one embodiment, said means for extraction of the metal cation is a perfusion fluid used in a dialysis or microdialysis system which further comprises the ligand exhibiting specific affinity for the target molecule.
According to the invention, the term “dialysis system” means any system allowing the passage of metal cations and/or at least one target molecule of interest from the extraction device through a dialysis 18 or microdialysis 2 membrane semipermeable to water and to the cations and/or molecules mentioned above.
The term “microdialysis system” is understood to mean a very small-scale dialysis system. For example, a microdialysis technique requires the insertion of a small microdialysis catheter, also called a microdialysis probe 1, into the tissue. The microdialysis probe is designed to mimic a blood capillary and consists of a tube with a semi-permeable membrane at its end, such as a hollow fiber membrane, which is connected to the inlet and outlet tubing. Microdialysis makes it possible to extract or deliver only the compounds capable of passing through a semi-permeable membrane whose cut-off threshold is selected according to the intended application. In the case of dialysis, this is often a dynamic diffusion phenomenon, guided by the difference in concentration of the diffusing species between each side of the dialysis membrane.
When using dialysis systems to extract compounds in low concentrations (metal cations and/or target molecules), the driving force is often quickly limited or saturated and the trapping of the compound(s) concerned is limited by the equilibrium concentration. Advantageously, a microdialysis system makes it possible to circumvent the problems of conventional chelating agents or ligands and to extract, locally or more generally, a very high proportion of the targeted metal cations (or of metal cations and target molecules), due to the maintaining inside the dialysis membrane of complexing chemical species (chelating agent(s) and/or ligand(s)) for at least one target metal cation and/or one target molecule. The chelating agents are advantageously grafted onto macromolecules or nanoparticles which have a mass greater than the cut-off threshold of the membrane, so that the complexing species remain within the perfusion fluid on one side of the dialysis membrane. Similarly, the ligands are advantageously present within or grafted onto macromolecules or nanoparticles which possess, or possess by their very nature, a mass greater than the cut-off threshold of the membrane. The dialysis system containing the complexing species is placed at the area of interest, for example at the brain (
Dialysis or microdialysis devices known to those skilled in the art may be used, on condition that they contain a semi-permeable dialysis membrane and a reservoir comprising a perfusion fluid containing at least one chelating agent and/or a ligand as mentioned above. As examples, devices which can be used are the medical devices developed by the companies M Dialysis AB, Sweden; Integra Life Sciences; in particular such as the microdialysis catheters (references 8010509, P000049, 8010337, this list not being exhaustive).
Dialysis systems, advantageously compact dialysis or microdialysis systems, can be used with an exchange reservoir of fixed volume 22 or 23. The specific capture of the cations to be extracted due to the chelating agent and of the target molecule due to the ligand makes it possible to maintain a purification gradient between the biological fluids to be purified and the perfusion fluid, even for small volumes of fluids with no circulation. In such an embodiment, the components of the biological fluids which one wishes to purify are preserved.
According to one embodiment, the dialysis system comprises a dialysis membrane 18 and a reservoir with a fixed volume 23, preferably with no circulation of fluid. Advantageously, the reservoir has a volume of less than 100 ml, preferably less than 20 ml, and more preferably less than 10 ml. Such a dialysis system is particularly suitable for a biological fluid such as cerebrospinal fluid.
Advantageously, and according to a preferred embodiment, the dialysis or microdialysis system comprises a microdialysis probe 1 continuously perfused with a perfusion fluid in the form of an aqueous solution (perfusate) which resembles the composition (ionic and/or molecular) of the surrounding biological fluid, at a low flow rate of less than 1 mL/min and preferably less than 0.1 mL/min. According to another embodiment, the means for extraction comprises a dialysis probe 1 continuously perfused with a perfusion fluid at a flow rate of less than 10 mL/min and preferably between 1 and 5 mL/min.
In one embodiment illustrated in
The microdialysis probe is preferably a linear or concentric probe. According to one embodiment illustrated in
In another embodiment illustrated in
In another embodiment, illustrated in
According to an advantageous embodiment compatible with any one of the preceding embodiments, the reservoirs 7/8, 22 or 23 are comprised in a housing 6 further comprising at least one of the following elements:
According to a preferred embodiment, the perfusion fluid comprises: i) a solution of nanoparticles comprising as active ingredient at least one chelating agent, and ii) a solution of at least one ligand exhibiting specific affinity for the target molecule, the average diameter of said nanoparticles and of the ligand being greater than the pores of the dialysis 18 or microdialysis 2 membrane. In one aspect, the cut-off threshold of the porous dialysis 18 or microdialysis 2 membrane is less than the mass of the chelating agent, i.e. the mass of the nanoparticle comprising at least one chelating agent.
Alternatively, the perfusion fluid comprises a solution of polymers, said polymers being grafted with at least one active ingredient which is a chelating agent, and a solution of at least one ligand exhibiting specific affinity for the target molecule, the average diameter being greater than the pores of said dialysis or microdialysis membrane. In this aspect, the cut-off threshold of the dialysis 18 or microdialysis 2 membrane is less than the mass of the chelating agent, i.e. the mass of the polymer onto which at least one chelating agent is grafted.
According to the invention, the term “solution” means a mixture of liquid and solid particles which remain evenly dispersed, the particles often being sufficiently small (microscopic or nanoscopic) for the mixture to remain stable and homogeneous.
According to one advantageous embodiment, the perfusion fluid is an “artificial cerebrospinal fluid” type of liquid comprising chelating agents based on polysiloxane and/or chitosan. Advantageously, it contains about 1 to 10 millimoles per liter of chelating agents of type EDTA, DTPA, and DOTA. According to a preferred embodiment, it may be one of the MetAEx® or proMetAEx® solutions marketed by Mexbrain.
According to one embodiment, said average diameter is greater than the pores of said dialysis or microdialysis membrane by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.
According to the invention, the term “average diameter” means the harmonic mean of the diameters of the compounds, in particular of nanoparticles or of polymers, of ligands. The compound size distribution is for example measured using a commercial particle size analyzer, such as a Malvern Zetasizer Nano-S particle size analyzer based on PCS (Photon Correlation Spectroscopy) which is characterized by an average hydrodynamic diameter. A method of measuring this parameter is also described in the 1996 ISO 13321 standard.
In one embodiment, the solution contains more than 1% by mass of nanoparticles or polymers, in particular more than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and preferably more than 10% by mass.
According to one embodiment, the chelating agent can be grafted onto said ligand specific for the target molecule. The chelating agents can thus be directly attached to the ligands, for example by covalence and covalent bonds, and more particularly by peptic bonds.
The nanoparticles which can be used in the extraction device advantageously comprise two characteristics:
In one embodiment, the nanoparticle comprises, as active ingredient, at least one chelating agent capable of complexing the metal cations, said chelating agent having a complexing constant log(KC1) for at least one of said metal cations that is greater than 10, and preferably greater than or equal to 15.
According to the invention, the term “silica-based nanoparticles” means nanoparticles characterized by a mass percentage of silicon of at least 8%.
According to the invention, the term “polysiloxane-based nanoparticles” means nanoparticles characterized by a mass percentage of silicon of at least 8%.
According to the invention, the term “polysiloxane” means an inorganic crosslinked polymer consisting of a chain of siloxanes.
The structural units of polysiloxane, which are identical or different, are of the following formula:
Si(OSi)nR4-n
where:
As a preferred example, the term “polysiloxane” includes in particular the polymers resulting from condensation of tetraethyl orthosilicate (TEOS) and of aminopropyltriethoxysilane (APTES) by a sol-gel process.
Advantageously, said nanoparticle thus comprises:
In one embodiment, the nanoparticles which can be used according to the invention do not comprise metal elements. In other words, in the above definition, said nanoparticle comprises only elements a. (polysiloxanes or silicon) and b. (chelating agents).
In one embodiment, the chelating agents complex the cations of metals Cu, Fe, Zn, Hg, Cd, Pb, Mn, Al, Ca, Mg, Gd.
In one embodiment, the chelating agents are obtained by grafting (covalent bond) onto the nanoparticle of one of the following complexing molecules or its derivatives, such as aminopolycarboxylic acids and their derivatives, in particular selected among: DOTA (1,4,7,10-tetraazacyclododecane-N,N′,N″,N′″-tetraacetic acid), DTPA (diethylenetriaminepentaacetic acid), D03A-pyridine of formula (I) below:
EDTA (2,2′,2″,2′″-(ethane-1,2-diyldinitrilo)tetraacetic acid), EGTA (ethylene glycol-bis(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid), BAPTA (1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTAGA ((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)pentanedioic acid), DFO (deferoxamine), amide derivatives such as for example DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10 tetraazacyclododecane) or NOTAM (1,4,7-tetrakis(carbamoylmethyl)-1,4,7-triazacyclononane), as well as mixed carboxylic acid/amide derivatives, phosphonic derivatives such as for example DOTP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(methylene phosphonate)) or NOTP (1,4,7-tetrakis(methylene phosphonate)-1,4,7-triazacyclononane), cyclam derivatives such as TETA (1,4,8,11-tetraazacyclotetradecane-N,N′,N″,N′″-tetraacetic acid), TETAM (1,4,8,11-tetraazacyclotetradecane-N,N′,N″,N′″-tetrakis(carbamoylmethyl)), TETP (1,4,8,11-tetraazacyclotetradecane-N,N′,N″,N′″-tetrakis(methylene phosphonate)), or mixtures thereof.
Preferably, said chelating agents above are linked directly or indirectly by covalent bond to the silicons of the polysiloxanes of the nanoparticle. The term “indirect” bond means the presence of a molecular “linker” or “spacer” between the nanoparticle and the chelating agent, said linker or spacer being covalently linked to one of the components of the nanoparticle.
According to a preferred embodiment, said nanoparticle is a polysiloxane-based nanoparticle with an average hydrodynamic diameter of between 3 and 100 nm, comprising the chelating agent obtained by grafting DOTA, DOTAGA, EDTA, or DTPA onto the nanoparticle.
According to a preferred embodiment, said nanoparticle is a nanoparticle with an average diameter greater than 20 kDa and less than 1 MDa, comprising the chelating agent obtained by grafting DOTA, DOTAGA, EDTA, or DTPA onto the nanoparticle.
According to a preferred embodiment, said solution comprising said nanoparticles also contains trace elements selected among Calcium, Magnesium, Iron, Copper, Zinc, or Manganese.
In another embodiment of the invention, polymers may be used in place of the aforementioned nanoparticles. In such case, said polymers are grafted with at least one chelating agent.
The term “polymer” is understood to mean any macromolecule formed from the covalent chaining of a very large number of repeating units which derive from one or more monomers. The polymers preferably used are, for example, from the family of chitosans, polyacrylamides, polyamines or polycarboxylics, polyethylene glycols, polyvinyl alcohols (PVA). For example, they can be polymers containing amino groups such as chitosan. According to a preferred embodiment, said polymer is biocompatible.
In one embodiment, the chelating agents or their derivatives grafted onto said polymers are aminopolycarboxylic acids and their derivatives, in particular selected among: DOTA, DTPA, DO3A-pyridine of formula (I) above, EDTA, EGTA, BAPTA, NOTA, DOTAGA, DFO, DOTAM, NOTAM, DOTP, NOTP, TETA, TETAM, and TETP or mixtures thereof.
Preferably, said chelating agents above are linked directly or indirectly by covalent bond to the polymer or to a polymer chain of more than 10 kDa and preferably more than 100 kDa. The term “indirect” bond means the presence of a molecular “linker” or “spacer” between the polymer and the chelating agent, said linker or spacer being covalently linked to one of the components of said polymer.
In one embodiment, the chelating agents or their derivatives grafted onto said polymers will comprise dithiocarbamate functional groups.
According to a preferred embodiment, said polymer grafted with a chelating agent is selected among: chitosan grafted with DPTA-BA or chitosan grafted with DFO or chitosan grafted with EDTA-BA or chitosan grafted with DOTAGA-A.
According to a preferred embodiment, said solution comprising said polymers also contains trace elements, selected among Calcium, Magnesium, Iron, Copper, Zinc, or Manganese.
Alternatively, the perfusion fluid is a solution of chelating molecules. Said chelating molecules may have an average diameter greater than the pores of said dialysis or microdialysis membrane, i.e. greater than the cutoff threshold of the membrane, in order to be retained within the liquid of the dialysis membrane. In another embodiment, they may have an average diameter smaller than the pores of said dialysis or microdialysis membrane, and in this case they can pass through the pores of the membrane before passing into the body and be naturally eliminated by the kidneys or liver.
The invention may find applications in particular in maintaining homeostasis, particularly in maintaining the homeostasis of two target compounds such as a metal cation and a target molecule.
According to a preferred embodiment, the device for the joint extraction of at least one metal cation and at least one target molecule of a biological fluid or of a biological aggregate, mentioned above, is used in the treatment of a disease selected among:
According to a preferred embodiment, the device for the joint extraction of at least one metal cation and at least one target molecule of a biological aggregate mentioned above is used to slow down the formation of, dissociate, or dissolve a biological aggregate, preferably in the form of oligomers, fibrils, or plaques comprising at least the target molecule; it is used in diagnosis, prevention, and/or therapy.
The invention also relates to a method for extracting metal cations and target molecules in a subject, comprising the administration of an implant onto which is grafted at least one chelating agent, or the use of a perfusion fluid containing at least one chelating agent within a device such as those mentioned above.
According to the invention, said “subject” is understood to mean a human or an animal in which prevention or treatment is to take place.
The invention is not limited to the preceding description, but encompasses all variants conceivable to a person skilled in the art within the framework of the protection sought.
For all appropriate purposes, the following patent document(s) is (are) cited:
For all appropriate purposes, the following non-patent element(s) is (are) cited:
| Number | Date | Country | Kind |
|---|---|---|---|
| 19 00699 | Jan 2019 | FR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/FR2020/050104 | 1/24/2020 | WO | 00 |
