Medical use of functionalized polymers

Abstract

The present disclosure relates to functionalized polymers for use in the treatment of diseases associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity in a subject in need thereof wherein the functionalized polymers are soluble in an aqueous solution, having a weight average molecular weight of 30 kDa to 5000 kDa, and wherein a portion of the monomer units are functionalized with chelating moieties, wherein a therapeutically effective amount of the functionalized polymer is administered orally to the subject.

Description

Technical Field

[0001] The present invention relates to medical uses of functionalized polymers. In particular, the present invention relates to functionalized polymers for treating diseases associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity in a subject in need thereof, wherein the functionalized polymer is soluble in aqueous solution, has a weight average molecular weight of 30 kDa to 5000 kDa, and wherein a portion of the monomer units are functionalized with a chelating moiety, wherein a therapeutically effective amount of the functionalized polymer is orally administered to the subject. Background Art

[0002] Heavy metals are prevalent in our environment due to human industrialization (waste disposal, factory pollution, nuclear reactions, etc.), and humans are inevitably exposed to these heavy metals due to polluted air, water and food (animals, crops, etc.). Lead and cadmium are found in trace amounts in the blood of humans throughout the world, and dietary intake has been directly linked to a number of serious conditions, including cancer and chronic kidney disease (CKD) (Satarug, S.; Vesey, D. A; Gobe, G. C. Health Risk Assessment of Dietary Cadmium Intake: Do Current Guidelines Indicate How Much Is Safe? Environ Health Perspect 2017, 125(3), 284–288; Satarug, S.; C. Gobe, G.; A. Vesey, D.; Phelps, K. R. Cadmium and Lead Exposure, Nephrotoxicity, and Mortality. Toxics 2020, 8(4), 86; European Food Safety Authority. Cadmium Dietary Exposure in the European Population. EFSA Journal No. 2012; 10(1):2551; Boon, P. E.; Pustjens, A. M.; Te Biesebeek, JD; Brust, GMH; Castenmiller, JJMDietary Intake and RiskAssessment of Elements for 1-and 2-Year-Old Children in the Netherlands. Food and Chemical Toxicology 2022, 161, 112810).

[0003] High blood concentrations of heavy metals, especially lead and cadmium, are most common in people who are exposed to high levels of heavy metals while working in mines and factories. However, even low exposures can lead to significant increases in blood concentrations over time. Once these heavy metals enter the body, most are eliminated through natural excretion pathways, but a small portion is absorbed into the blood and accumulates in organs (liver, kidneys, bones) over a long period of time. This long-term exposure to heavy metals has been found in different regions, especially in children, due to contaminated food and water. Equally worrying is the vicious cycle of heavy metals in the elderly or in people with certain medical conditions, such as chronic kidney disease (CKD). The natural elimination pathways in these populations are less efficient, so the heavy metals that accumulate in these systems are greater in quantity and are more toxic.

[0004] These metals and other heavy metals have been linked to a variety of conditions, including but not limited to neurodegeneration and neurodegenerative diseases such as Parkinson's disease, cardiovascular disease, renal failure and chronic kidney disease (CKD), male fertility impairment, miscarriage, and childhood stunting (Reuben, A. Childhood Lead Exposure and Adult Neurodegenerative Disease. JAD 2018, 64(1), 17–42; Kumar, A.; Kumar, A.; MMs, C.-P.; Chaturvedi, A. K.; Shabnam, A. A.; Subrahmanyam, G.; Mondal, R.; Gupta, D. K.; Malyan, S. K.; Kumar, S. S.; A. Khan, S.; Yadav, K. K. Lead Toxicity: Health Hazards, Influence on Food Chain, and Sustainable Remediation Approaches. International Journal of Environmental Research and Public Health 2020, 17(7), 2179; Leggett, R. W. An Age-SpecificKinetic Model of Lead Metabolismin Humans.Environ Health Perspect 1993,101(7),598–616; Rossi, E.Low Level Environmental Lead Exposure–A ContinuingChallenge.Clin Biochem Rev 2008,29(2),63–70).

[0005] At the cellular level, these metals use their strong affinity for sulfur, nitrogen, and oxygen to bind and inhibit a variety of enzymes, proteins, and cell signaling pathways, while increasing the concentration of reactive oxygen species (ROS) (Chen, P.; Bornhorst, J.; Diana Neely, M.; Avila, D.S. Mechanisms and Disease Pathogenesis Underlying Metal-Induced Oxidative Stress. Oxidative Medicine and Cellular Longevity 2018, 2018, 1–3; Ercal, N.; Gurer-Orhan, H.; Aykin-Burns, N. Toxic Metals and Oxidative Stress Part 1: Mechanisms Involved in Metal Induce Oxidative Damage. Current Topics in Medicinal Chemistry. 1st ed. 2001, pp 529–539; Flora, S. J. S.; Mittal, M.; Mehta, A. Heavy Metal Induced Oxidative Stress & Its Possible Reversal by Chelation Therapy. INDIAN J MED RES 2008, 24). Globally, these metals are highly nephrotoxic because they are directly related to a decrease in glomerular filtration rate (GFR) and accumulate in large quantities in organs after being absorbed in the blood (Reyes, JL; Molina-Jijón, E.; R.; Bautista-García, P.; Debray-García, Y.; Namorado, M. del C. Tight Junction Proteins and Oxidative Stress in HeavyMetals-Induced Nephrotoxicity. BioMed Research International 2013, 2013, 1–14; Johri, N.; Jacquillet, G.; Unwin, R. Heavy Metal Poisoning: The Effects of Cadmium on the Kidney.Biometals:an international journal on the role of metal ions inbiology,biochemistry,and medicine 2010,23,783–792;Lentini,P.;Zanoli,L.;Granata,A.;Signorelli,SS;Castellino,P.;Dellaquila,R.Kidney and HeavyMetals-The Role of Environmental Exposure.Molecular Medicine Reports 2017, 15(5), 3413–3419). During osteoporosis and similar conditions associated with the aging anatomical system, accumulation within the bone can reintroduce these metals into the blood.

[0006] Current treatments for acute heavy metal poisoning (>200 μg / L) involve administration of chelating moieties, also known as chelators, such as EDTA and DMSA. However, this approach is not applicable to lower blood concentration levels (<100 μg / L) due to low efficiency. Trace amounts of lead (<50 μg / L) and cadmium (<1 μg / L) have been associated with occult symptoms in humans, although these values ​​are considered "acceptable" and do not require treatment strategies defined by the WHO and CDC. The only treatment strategy proposed for this population is to move away from the source of exposure (ABLES-Reference Blood Lead Levels(BLLs) for Adults in the US|NIOSH|CDC. https: / / www.cdc.gov / niosh / topics / ables / ReferenceBloodLevelsforAdults.html(accessed 2022-05-23); Center for Disease Control(CDC). ReferenceValue|Lead|CDC.https: / / www.cdc.gov / nceh / lead / data / blood-lead-reference-value.htm(accessed 2022-05-19); World Health Organization; Regional Office for Europe; Joint WHO / Convention Task Force on the Health Aspects of AirPollution.Health Risks of Heavy Metals from Long-Range Transboundary AirPollution; World Health Organization Regional Office Europe: Copenhagen, 2007).

[0007] Therefore, it is very difficult to stay away from the source of exposure to these harmful substances when they come from food and water.

[0008] WO2019 / 122790 discloses a medical device for maintaining metal homeostasis that can be placed in the body for therapeutic purposes, which comprises a chelating part for extracting metals.

[0009] WO2022 / 023677 describes statistical polysaccharides with a weight-average molecular weight of 100 kDa-1000 kDa and their use in dialysis processes for capturing at least one metal during MRI imaging, during brachytherapy or during marking of food to prevent counterfeiting.

[0010] Therefore, it remains a challenge to design a safe and effective therapeutic approach to meet the clinical need for treating diseases associated with heavy metal-induced toxicity or carcinogenicity in subjects in need thereof.

[0011] Brief description of the invention

[0012] A first aspect of the present disclosure relates to a functionalized polymer for use in treating a disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity in a subject in need thereof, wherein the functionalized polymer is soluble in aqueous solution, has a weight average molecular weight of 30 kDa-5000 kDa, and wherein a portion of the monomer units are functionalized with a chelating moiety, wherein a therapeutically effective amount of the functionalized polymer is orally administered to the subject.

[0013] A second aspect of the present disclosure relates to an oral formulation comprising a functionalized polymer as defined above and one or more pharmaceutically acceptable excipients, wherein the unit dose of the functionalized polymer is 0.1 mg-500 mg, preferably 1 mg-100 mg, more preferably 2 mg-10 mg, even more preferably about 5 mg. DETAILED DESCRIPTION OF THE INVENTION

[0015] definition

[0016] Hereinafter, the terms used herein are defined with their meanings.

[0017] The term "about" or "around" has the following meaning in this document: the following values ​​may vary by ±20%, preferably ±10%, more preferably ±5%, even more preferably ±2%, even more preferably ±1%.

[0018] Unless otherwise defined, "%" herein has the meaning of weight percent (wt %), also referred to as weight percent by weight (w / w %).

[0019] Unless otherwise indicated, the terms "treating" or "treatment" as used herein mean reversing, alleviating, inhibiting the progression of a disorder or condition to which the term applies, or preventing, alleviating a disorder or condition to which the term applies, or reversing, alleviating, inhibiting the progression of one or more symptoms of a disorder or condition to which the term applies, or preventing one or more symptoms of a disorder or condition to which the term applies. More preferably, in the context of the present invention, "treatment" may include reducing heavy metals in the blood of a subject, or preventing the increase of certain heavy metals in the blood due to the entry of heavy metals into the blood circulation from water or food.

[0020] As used herein, the term "prevent" refers to at least significantly limiting the absorption of certain heavy metals in the blood of a subject. For example, significantly limiting means that the absorption of heavy metals in a subject exposed to heavy metals is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or even 100% compared to a subject not treated with the functionalized polymer of the present disclosure and exposed to heavy metals.

[0021] As used herein, the terms "heavy metals", "heavy metals" and "heavy metal ions" are used interchangeably to refer to metal atoms and cations that are toxic to humans. These toxic metals are completely inactive in biological systems and pose a threat to the health and function of the system even at trace levels. Such threats include enzyme and protein inhibition, oxidation of cell membranes and degradation of signal transduction pathways, inhibition of DNA repair and promotion of DNA degradation, apoptosis, mutagenesis, etc., leading to cardiovascular disease, neurodegeneration and cognitive impairment, renal dysfunction, cancer and death. For example, the heavy metal can be selected from the group consisting of lead (Pb), cadmium (Cd), arsenic (As), chromium (Cr), mercury (Hg), cobalt (Co), aluminum (Al), antimony (Sb), barium (Ba), bismuth (Bi), gallium (Ga), germanium (Ge), gold (Au), indium (In), nickel (Ni), platinum (Pt), silver (Ag), strontium (Sr), tellurium (Te), thallium (Tl), tin (Sn), titanium (Ti), vanadium (V), plutonium (Pu), cesium (Cs), cerium (Ce), zirconium (Zr), ruthenium (Ru), technetium (Tc), radium (Ra), thorium (Th), americium (Am), iridium (Ir), californium (Cf), polonium (Po) and uranium (U), or a mixture thereof, preferably selected from the group consisting of lead, cadmium, arsenic, chromium and mercury, more preferably lead or cadmium. In a specific embodiment, the heavy metal can be radioactive and can be selected from the following group: Co (60), U (232, 233, 234, 235, 236 or even 238), Pu (238, 239, 240, 241), Sr (90), Cs (135, 137), Ce (144), Zr (93, 95), Ru (106), Tc (99), Sn (126), Am (241), Ir (192), Po (210), Ra (226), Pu (238), Am (241), Cf (252).

[0022] As used herein, the term "chelator" or "chelating agent" or "chelating moiety" is used to define a chemical structure or part that exhibits a relatively high affinity for certain elements and displays at least two coordination sites. The affinity enables chelators to semi-permanently chelate metals and can eliminate any affinity they display with biomolecules. The main function of chelators is to "neutralize" heavy metals, which can be achieved by maintaining their chelation with heavy metals and preventing them from reacting and interacting with other biomaterials. Chelators can have a pincer-type structure or part, which has two or more relative parts, formed by chemical groups (sulfhydryl, keto, carboxyl, hydroxyl, etc.) or neutral charges (amino groups) that have a negative charge in a biological environment. These groups are appropriately spaced to comfortably accommodate metal ions in their structure.

[0023] In the context of the present disclosure, the chelating moiety may be:

[0024] DOTA: 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid;

[0025] NOTA: 1,4,7-triazacyclononane-1,4,7-triacetic acid;

[0026] NODAGA: 1,4,7-triazacyclononane-1-pentanedioic acid-4,7-diacetic acid;

[0027] DOTAGA: 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid;

[0028] DOTAM: 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclodecane;

[0029] NOTAM: 1,4,7-tetrakis(carbamoylmethyl)-1,4,7-triazacyclononane;

[0030] DOTP: 1,4,7,10-tetraazacyclododecane 1,4,7,10-tetrakis(methylenephosphonate);

[0031] NOTP: 1,4,7-tetrakis(methylenephosphonate)-1,4,7-triazacyclononane;

[0032] TETA: 1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid;

[0033] TETAM: 1,4,8,11-tetraazacyclotetradecane-N,N',N",N''-tetrakis(carbamoylmethyl);

[0034] DTPA: diethyltriaminepentaacetic acid;

[0035] Bz-DFO: benzyldeferoxamine;

[0036] DFO: deferoxamine.

[0037] As used herein, the term "effective amount" or "therapeutically effective amount" of a compound refers to an amount of a compound that causes a biological or medical response in a subject, such as improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease. In specific embodiments, such a therapeutically effective amount is an amount sufficient to prevent absorption of the heavy metal in the subject.

[0038] As used herein, with respect to the Rc moiety (i.e., the chelating moiety), the term "independently" means that each Rc of the functionalized statistical chitosan of formula (I) is a chelating moiety and may be different or the same from each other. For example, each Rc is the same, i.e., there is one type of Rc in the entire functionalized statistical chitosan, or there may be more than one type of Rc in the entire functionalized statistical chitosan, i.e., two, three, four, five or even n different Rc, n being an integer.

[0039] As used herein, the term "pharmaceutically acceptable excipient" refers to an inactive substance added with a drug and is part of a formulation mixture. Pharmaceutically acceptable excipients are, for example, fillers, solvents, diluents, carriers, adjuvants, dispersants and sensor, delivery agents, such as preservatives, disintegrants, wetting agents, emulsifiers, suspending agents, thickeners, sweeteners, flavoring agents, aromatics, antibacterial agents, fungicides, lubricants and extended delivery control agents, antioxidants, glidants. Their selection and appropriate proportions depend on the nature and mode of administration and the dosage.

[0040] As used herein, "combination" refers to a fixed combination in one dosage unit form, or a combined administration, wherein a compound of the present disclosure and a combination partner (e.g., another drug, also referred to as a "therapeutic agent" or "adjuvant") as described below) can be administered independently simultaneously or separately at time intervals, especially wherein these time intervals allow the combination partners to exhibit a cooperative effect, such as a synergistic effect. The individual components can be packaged in a kit or packaged separately. One or both components (e.g., powders or liquids) can be reconstituted or diluted to the desired dose prior to administration. As used herein, the terms "co-administration" or "combined administration" and the like encompass administration of selected combination partners to a single subject (e.g., patient) in need thereof, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

[0041] The terms "patient", "subject", "individual", etc. are used interchangeably herein and refer to humans. In some embodiments, patients, subjects, or individuals in need of treatment include people who already have a disease, condition, or disorder, i.e., a disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity.

[0042] Functionalized polymers for use thereof according to the present disclosure

[0043] A first aspect of the present disclosure relates to a functionalized polymer for use in treating a disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity in a subject in need thereof, wherein the functionalized polymer is soluble in aqueous solution, has a weight average molecular weight of 30 kDa-5000 kDa, and wherein a portion of the monomer units are functionalized with a chelating moiety, wherein a therapeutically effective amount of the functionalized polymer is orally administered to the subject.

[0044] In certain embodiments, the weight average molecular weight of the functionalized polymer is 100 kDa-1000 kDa, preferably 150 kDa-750 kDa, more preferably 200 kDa-600 kDa, even more preferably 250 kDa-400 kDa, even more preferably about 300 kDa.

[0045] In another embodiment, the weight average molecular weight of the functionalized polymer is 30 kDa-100 kDa, preferably 32 kDa-80 kDa, more preferably 34 kDa-60 kDa, even more preferably 35 kDa-50 kDa, even more preferably about 40 kDa.

[0046] In one embodiment, the functionalized polymer comprises at least 1 w% chelating moieties, such as 1 w%-40 w%, preferably 5 w%-30 w%, more preferably 10 w%-25 w%, even more preferably about 10 w% or 17 w% or 22 w%.

[0047] The chelating moiety can chelate one or more metals. Each chelating moiety can include two or more coordination sites. Preferably, the coordination site is a nitrogen or oxygen atom. Advantageously, each chelating moiety includes 4-8 coordination sites, more preferably 6-8 coordination sites, and even more preferably each chelating moiety includes 6 coordination sites.

[0048] As used herein, the term "coordination site" refers to a single functional group capable of complexing a metal. For example, an amine functional group represents a coordination site by forming a coordinate bond between the nitrogen atom and the metal, and a hydroxamic acid functional group also represents a coordination site by forming a coordinate bond between the oxygen of the carbonyl unit and by forming a covalent bond with the oxygen of the N-oxide unit, thus forming a five-membered ring.

[0049] In one embodiment, each chelating moiety is selected from the group consisting of DOTA (1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane-1-pentanedioic acid-4,7-diacetic acid), DOTAGA (2-(4,7,10-tris(carboxymethyl)-1,4 ,7,10-tetraazacyclododecane-1-yl)pentanedioic acid), DOTAM (1,4,7,10-tetra(carbamoylmethyl)-1,4,7,10-tetraazacyclodecane), NOTAM (1,4,7-tetra(carbamoylmethyl)-1,4,7-triazacyclononane), DOTP (1,4,7,10-tetraazacyclododecane 1,4,7,10-tetra(methylenephosphonate)), NOTP (1 ,4,7-tetrakis(methylenephosphonate)-1,4,7-triazacyclononane), TETA (1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid), TETAM (1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetrakis(carbamoylmethyl)), DTPA (diethyltriaminepentaacetic acid), Bz-DFO (benzyldeferoxamine) and DFO (deferoxamine) and mixtures thereof, preferably selected from the group consisting of DOTAGA, Bz-DFO, DFO, DOTAM and DTPA, and more preferably, the chelating moiety is DOTAGA, or DOTAM, or DTPA, or a mixture of DOTAGA and Bz-DFO, or a mixture of DOTAGA and NODAGA, or a mixture of DOTAGA and DTPA, or a mixture of DOTAGA and DOTAM.

[0050] In one embodiment, the chelating moiety is selected from the group consisting of:

[0051]

[0052]

[0053] In a specific embodiment, the heavy metal chelated by the chelated portion or a mixture thereof is selected from the group consisting of lead (Pb), cadmium (Cd), arsenic (As), chromium (Cr), mercury (Hg), cobalt (Co), aluminum (Al), antimony (Sb), barium (Ba), bismuth (Bi), gallium (Ga), germanium (Ge), gold (Au), indium (In), nickel (Ni), platinum (Pt), silver (Ag), strontium (Sr), tellurium (Te), thallium (Tl), tin (Sn), titanium (Ti), vanadium (V), plutonium (Pu), cesium (Cs), cerium (Ce), zirconium (Zr), ruthenium (Ru), technetium (Tc), radium (Ra), thorium (Th), americium (Am), iridium (Ir), californium (Cf), polonium (Po) and uranium (U), or a mixture thereof, preferably selected from the group consisting of lead, cadmium, arsenic, chromium and mercury, more preferably lead or cadmium.

[0054] In a specific embodiment, the heavy metal is radioactive, for example selected from the following group: Co (60), U (232, 233, 234, 235, 236 or even 238), Pu (238, 239, 240, 241), Sr (90), Cs (135, 137), Ce (144), Zr (93, 95), Ru (106), Tc (99), Sn (126), Am (241), Ir (192), Po (210), Ra (226), Pu (238), Am (241), Cf (252).

[0055] The functionalized polymer may be selected from the group consisting of polysaccharides, polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polyallylamine (PAH), wherein a portion of the monomer units are functionalized with a chelating moiety.

[0056] In a specific embodiment, the polysaccharide may be selected from digestible polysaccharides, such as starch; or non-digestible polysaccharides, such as chitosan, cellulose, chitin, β-glucan, xylan, pectin, mucilage, gum, lignin, galactan, agar, levan, fucoidan galactoglucan, sulfated polysaccharides or mixtures thereof, preferably chitosan. In a preferred embodiment, the polysaccharide is selected from non-digestible polysaccharides. In fact, non-digestible polysaccharides have β-glycosidic bonds and are more difficult to digest compared to digestible polysaccharides with α-glycosidic bonds, while digestible polysaccharides can be digested more quickly by α-amylase present in the digestive tract.

[0057] In one embodiment, the functionalized polymer is polyethylene glycol (PEG), preferably with DTPA or DOTAM or a mixture thereof as the chelating moiety, and more preferably wherein the weight average molecular weight of the polyethylene glycol is 30 kDa-100 kDa, preferably 32 kDa-80 kDa, more preferably 34 kDa-60 kDa, even more preferably 35 kDa-50 kDa, even more preferably about 40 kDa.

[0058] In a specific embodiment, the functionalized polymers of the present disclosure are selected from polymers that do not enter the blood after oral administration. As used herein, a polymer is considered to not enter the blood after oral administration when less than 5%, preferably less than 3%, and more preferably less than 1% of the polymer is in the blood after oral administration, which is typically determined by animal experiments as described in the Examples, i.e., using ICPMS to perform fluorescence quantification studies of gadolinium in a red filter.

[0059] In fact, without being bound by any theory, the inventors believe that the polymer acts locally by chelating a portion of the heavy metals present in water and / or food and / or air pollution, for example from pollution, contamination, accidents, war zones such as dirty bombs, by being retained in the gastrointestinal tract, before being absorbed, thus preventing them from passing the intestinal barrier. Therefore, the fact that the polymer does not pass the intestinal barrier allows a local action in the gastrointestinal tract and avoids the common adverse events of the presence of the chelated moiety in the systemic compartment as observed in prior art treatments.

[0060] In one embodiment, at least 90%, preferably at least 95%, more preferably at least 99% of said polymer is excreted in the feces within 7 days after oral administration to said subject.

[0061] In another embodiment, the polymers for use thereof according to the present disclosure are selected from polymers that degrade poorly in the gastrointestinal tract, and whose weight average molecular weight does not substantially change after they pass through the gastrointestinal tract.

[0062] Preferred embodiments of polymers: Functionalized PEG

[0063] In a specific embodiment, the functionalized polymer is polyethylene glycol (PEG), more preferably an eight-arm PEG amine according to the following formula (V):

[0064]

[0065] wherein n is 100-120, more preferably 105-115, even more preferably 109-113, even more preferably about 111.

[0066] In one embodiment, the eight-arm PEG of Formula V is a functionalized PEG, PEG-DTPA, of Formula (VI):

[0067]

[0068] wherein n is 100-120, more preferably 105-115, even more preferably 109-113, even more preferably about 111.

[0069] In another embodiment, the eight-arm PEG of Formula V is a functionalized PEG of Formula (VII), PEG-DOTAM, which has Formula (VII):

[0070]

[0071] wherein n is 100-120, more preferably 105-115, even more preferably 109-113, even more preferably about 111.

[0072] Preferred embodiment of polymer: functionalized statistical chitosan

[0073] In a specific embodiment, the functionalized polymer is chitosan, preferably functionalized chitosan, having a statistical macromolecular structure with a weight average molecular weight of 100 kDa to 1000 kDa and formula (I):

[0074]

[0075] in

[0076] Each Rc is a chelating moiety,

[0077] Each Z is independently a linker which is a single bond or a hydrocarbon chain containing 1 to 12 carbon atoms, which is linear or branched and optionally contains one or more unsaturated groups and can contain one or more heteroatoms, preferably selected from nitrogen, oxygen, sulfur and halogens,

[0078] x is 0.005-0.7, preferably 0.05-0.7,

[0079] y is 0.01-0.7, preferably 0.05-0.2,

[0080] The ratio y / x is greater than or equal to 0.05, preferably greater than or equal to 0.15, and

[0081] The sum of x+y is greater than or equal to 0.15, preferably greater than or equal to 0.30, more preferably greater than or equal to 0.35.

[0082] It should be understood that in the above formula I, there may be more than one group in the functionalized statistical chitosan Rc. These Rc groups may be the same or different from each other. Each is independently selected from a group carrying a chelating moiety. The same applies to Z-linkers, there may be multiple Z-linkers, and they may be the same or different from each other.

[0083] In one embodiment, in Formula I, x is 0.005-0.6; y is 0.1-0.9; the ratio y / x is greater than 0.16; and the sum of x+y is greater than 0.30.

[0084] In a specific embodiment, the functionalized statistical chitosan of the present disclosure has at least 10 15 The complexation constant.

[0085] In one embodiment, the functionalized statistical chitosan of formula I is a functionalized statistical chitosan of formula (II):

[0086]

[0087] in

[0088] Rc1 and Rc2 are different chelating moieties,

[0089] Z1 and Z2, identical or different, are linkers which are single bonds or hydrocarbon chains containing 1 to 12 carbon atoms, which are linear or branched and optionally contain one or more unsaturated groups and one or more heteroatoms, which are preferably selected from nitrogen, oxygen, sulfur and halogens,

[0090] x is 0.005-0.7, preferably 0.05-0.7, and more preferably 0.2-0.6,

[0091] y=z+w is 0.01-0.7, preferably 0.05-0.2,

[0092] The ratio y / x is greater than or equal to 0.05, preferably greater than or equal to 0.15,

[0093] The sum of x+y is greater than or equal to 0.15, preferably greater than or equal to 0.30, more preferably greater than or equal to 0.35, and

[0094] z / y is 0.5-1.

[0095] In this embodiment, the functionalized statistical chitosan of formula (II) may comprise a single type of chelating moiety Rc1 when z equals 1, or 2 types of moieties, including chelating moieties Rc1 and Rc2, when 0.5≤z<1.

[0096] In one embodiment, z / y is from 0.8 to 0.99, so that the Rc1 moiety is in the majority.

[0097] In another embodiment, in Formula II, x is 0.005-0.6; y is 0.1-0.9; the ratio y / x is greater than 0.3; and z is 0.5-1.

[0098] Rc part (Rc, Rc1 and Rc2)

[0099] As used herein, the term "Rc moiety" refers to the Rc moiety of Formula I, and when Rc2 is present, the term "Rc1 and Rc2 moieties" refers to Rc1 and Rc2 of Formula II. According to the present disclosure, the Rc1 and Rc2 groups are chelating moieties. In other words, the Rc, Rc1 and Rc2 moieties are capable of chelating one or more metals by forming a complex.

[0100] Each Rc, Rc1 and Rc2 part may contain two or more coordination sites. Preferably, the coordination sites are nitrogen or oxygen atoms. Advantageously, each Rc, Rc1 and Rc2 part contains 4-8 coordination sites, more preferably 6-8 coordination sites, and even more preferably each Rc, Rc1 and Rc2 part contains 6 coordination sites.

[0101] As used herein, the term "coordination site" refers to a single functional group capable of complexing a metal. For example, an amine functional group forms a coordination site by forming a coordinate bond between the nitrogen atom and the metal, and a hydroxamic acid functional group also forms a coordination site by forming a coordinate bond at the oxygen of the carbonyl unit and by forming a covalent bond with the oxygen of the N-oxide unit, thus forming a five-membered ring.

[0102] In one embodiment, for the functionalized statistical chitosan of formula I, each Rc moiety is independently selected from the group consisting of DOTA (1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane-1-pentanedioic acid-4,7-diacetic acid), DOTAGA (2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)pentanedioic acid), DOTAM (1,4,7,10-tetra(carbamoylmethyl)-1,4,7,10-tetraazacyclodecane), NOTAM (1,4,7-tetra(carbamoylmethyl)-1,4,7 -triazacyclononane), DOTP (1,4,7,10-tetraazacyclododecane 1,4,7,10-tetra(methylenephosphonate)), NOTP (1,4,7-tetra(methylenephosphonate)-1,4,7-triazacyclononane), TETA (1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetrachloroacetic acid), TETAM (1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetra(carbamoylmethyl)), DTPA (diethyltriaminepentaacetic acid), Bz-DFO (benzyldeferoxamine) and DFO (deferoxamine), preferably selected from the group consisting of DOTAGA, Bz-DFO, DFO, DOTAM and DTPA, more preferably the Rc group is DOTAGA.

[0103] In another embodiment, for the functionalized statistical chitosan of formula II, Rc1 and Rc2 are independently selected from the group consisting of DOTA, NOTA, NODAGA, DOTAGA, DOTAM, NOTAM, DOTP, NOTP, TETA, TETAM, DTPA, Bz-DFO and DFO, preferably selected from the group consisting of DOTAGA, Bz-DFO, DFO, DOTAM and DTPA.

[0104] In one embodiment, the chelating moiety is selected from the group consisting of:

[0105]

[0106]

[0107] In one embodiment, for the functionalized statistical chitosan of formula I, the group Rc is DOTAGA, and preferably, z / y=1.

[0108] In another embodiment, for the functionalized statistical chitosan of formula II, the group Rc1 is DOTAGA and the group Rc2 is Bz-DFO.

[0109] Z linkers (Z, Z1 and Z2)

[0110] As used herein, the term "Z linker" refers to the Z linker of Formula I, and when a Z2 linker is present, the term "Z1 and Z2 linkers" refers to the Z1 and Z2 linkers of Formula II.

[0111] The choice of Z, Z1 and Z2 linkers in formulae I and II depends mainly on the Rc, Rc1 and Rc2 moieties and the metal to be chelated. In practice, in particular, due to stearic acid, the Rc, Rc1 and Rc2 moieties can be more or less close to the nitrogen 6-membered ring of the glucosamine unit.

[0112] In formula I, each Z is independently a linker which is a single bond or a hydrocarbon chain containing 1 to 12 carbon atoms, which is linear or branched and optionally contains one or more unsaturations and one or more heteroatoms, preferably selected from nitrogen, oxygen, sulfur and halogens.

[0113] In one embodiment, in Formula I, each Z is independently selected from the group consisting of a bond, a linear or branched alkyl chain having 1 to 12 carbon atoms, and a linear or branched alkenyl chain having 2 to 12 carbon atoms, the alkyl and alkenyl chains being interrupted by one or more C6-C10 aryl groups and / or by one or more heteroatoms or groups selected from the group consisting of -O-, -S-, -C(O)-, -NR'-, -C(O)NR'-, -NR'-C(O)-, -NR'-C(O)-NR'-, -NR'-C(O)-O-, -OC(O)NR', ​​-C(S)NR'-, -NR'-C(S)-, -NR'-C(S)-NR, the alkyl and alkenyl chains being substituted by one or more groups selected from the group consisting of halogen, -OR', -COOR', -SR', -NR'2, and each R' is independently H or C1-C6 alkyl.

[0114] Advantageously, in formula I, each Z is independently selected from the group consisting of a bond and a linear or branched alkyl chain having 1 to 12 carbon atoms, which alkyl chain may be interrupted by one or more C6-C10 aryl groups and / or by one or more heteroatoms or groups selected from the group consisting of -O-, -S-, -C(O)-, -NR'-, -C(O)NR'-, -NR'-C(O)-, -C(S)NR'-, -NR'-C(S)-NR', and each R' is independently H or a C1-C6 alkyl group.

[0115] In one embodiment, each Z is an alkyl chain having 1 to 12 carbon atoms.

[0116] In another embodiment, each Z is polyethylene glycol (PEG).

[0117] Advantageously, in formula II, Z1 and Z2 are independently a single bond or a hydrocarbon chain having 1 to 12 carbon atoms, wherein the chain may be linear or branched and may have one or more unsaturations and may have one or more heteroatoms, preferably selected from nitrogen, oxygen, sulfur and halogens.

[0118] In one embodiment, in Formula II, Z1 and Z2 are independently selected from the group consisting of a bond, a straight or branched alkyl chain having 1 to 12 carbon atoms, and a straight or branched alkenyl chain having 2 to 12 carbon atoms, wherein the alkyl and alkenyl chains may be interrupted by one or more C6-C10 aryl groups and / or by one or more heteroatoms or groups selected from the group consisting of -O-, -S-, -C(O)-, -NR'-, -C(O)NR'-, -NR'-C(O)-, -NR'-C(O)-NR'-, -NR'-C(O)-O-, -OC(O)NR', ​​-C(S)NR'-, -NR'-C(S)-, -NR'-C(S)-NR, wherein the alkyl and alkenyl chains may be substituted by one or more groups selected from the group consisting of halogen, -OR', -COOR', -SR', -NR'2, and each R' is independently H or C1-C6 alkyl.

[0119] In one embodiment, in Formula II, Z1 and Z2 are independently selected from the group consisting of a bond and a straight or branched alkyl chain having 1 to 12 carbon atoms, wherein the alkyl chain may be interrupted by one or more C6-C10 aromatic groups and / or by one or more heteroatoms or groups selected from the group consisting of -O-, -S-, -C(O)-, -NR'-, -C(O)NR'-, -NR'-C(O)-, -C(S)NR'-, -NR'-C(S)-NR', and each R' is independently H or C1-C6 alkyl.

[0120] In a specific embodiment, Z1 and / or Z2 is an alkyl chain having 1 to 12 carbon atoms.

[0121] In another specific embodiment, Z1 and / or Z2 is polyethylene glycol (PEG).

[0122] Functionalized statistical chitosan monomer units

[0123] The functionalized statistical chitosan in Formula I of the present disclosure consists of three different monomer units, namely N-acetylglucosamine A-type unit, glucosamine B-type unit and glucosamine C-type unit, which are functionalized by a chelating moiety (Rc type) that is linked to the nitrogen of glucosamine via a linker (Z type).

[0124] The functionalized statistical chitosan is a statistical polymer. In other words, the order of the individual monomer units A, B and type C is random.

[0125] The functionalized statistical chitosan in formula II of the present disclosure is composed of 4 different monomer units, namely N-acetylglucosamine A-type unit, glucosamine B-type unit and two glucosamine C-type units, namely C1 and C2, which are functionalized by a chelating part (Rc1 type or Rc2 type) connected to the nitrogen of glucosamine through a linker (Z1 type or Z2 type).

[0126] The functionalized statistical chitosan in formula II is a statistical polymer. In other words, the order of the individual monomer units A, B, C1 and C2 is random.

[0127] In formula I and II, x represents the proportion of A units, and x is 0.005-0.7, preferably 0.05-0.7, more preferably 0.2-0.6, even more preferably x is 0.25-0.4, typically about 0.3. In one embodiment, x is 0.025-0.075, more preferably 0.04-0.06, typically about 0.05.

[0128] In formula I and II, y represents the proportion of C-type units, and y is 0.01-0.7, preferably 0.05-0.2. In one embodiment, y is 0.03-0.2, preferably 0.05-0.1, even more preferably 0.07-0.08, typically about 0.072. In another embodiment, y is 0.05-0.3, more preferably 0.1-0.2, typically about 0.15 or 0.12.

[0129] In another embodiment, when Rc1 and Rc2 of formula II are independently selected from the group consisting of DOTA, NOTA, NODAGA, DOTAGA, DOTAM, NOTAM, DOTP, NOTP, TETA, TETAM, DTPA, Bz-DFO and DFO, preferably selected from the group consisting of DOTAGA, Bz-DFO, DFO, DOTAM and DTPA, and y is 0.05-0.3, more preferably 0.1-0.2, typically about 0.12, Rc1 is 0.06-0.08, typically about 0.07, and Rc2 is 0.04-0.06, typically about 0.05.

[0130] The remaining monomer units in formulae I and II are B units. Therefore, in formulae I and II, the ratio of B units is equal to 1-xy.

[0131] According to the present disclosure, in formula I and II, the ratio y / x is greater than or equal to 0.05, preferably greater than or equal to 0.15. In fact, the effectiveness of functionalized statistical chitosan is determined by the number of chelation sites, which is directly related to the amount of metal required, for example, for reducing local inflammation caused by metal homeostasis disorders and / or for inducing local inflammation of metal homeostasis disorders in subjects in need, and for reducing oxidative stress.

[0132] In order to be administered as a solution to a subject in need thereof and at the same time be effective, the functionalized statistical chitosan must be soluble at physiological pH, i.e. pH 4.8-8. For this purpose, the sum of x+y may be greater than or equal to 0.15, preferably greater than or equal to 0.30, more preferably greater than or equal to 0.35.

[0133] The combination of the specific ratio of the number of A units to the number of C units and the sum of the ratio of A units to the ratio of C units enables sufficient chelation and solubility to allow the functionalized statistical chitosan to be used to treat diseases characterized by local inflammation-induced and / or induced metal homeostasis disorders and increased oxidative stress in subjects in need thereof.

[0134] According to the present invention, z / y is 0.5 to 1. In other words, the C-type unit may be only a unit containing Z1 as a linker and Rc1 as a group containing a chelating moiety.

[0135] The weight average molecular weight of the functionalized statistical chitosan is 100 kDa-1000 kDa, preferably 150 kDa-750 kDa, more preferably 200 kDa-600 kDa, even more preferably 250 kDa-400 kDa, even more preferably about 300 kDa.

[0136] In one embodiment, the functionalized statistical chitosan is selected from the following functionalized statistical chitosans:

[0137] - functionalized statistical chitosan of formula II, wherein z / y=1, Rc1 is DOTAGA, and Z1 is a bond;

[0138] - functionalized statistical chitosan of formula II, wherein z / y=1, Rc1 is DTPA and Z1 is a bond; and

[0139] - Functionalized statistical chitosan of formula II, wherein 0.5≤z / y≤1, Rc1 is DOTAGA, Z1 is a bond, Rc2 is Bz-DFO, Z2 is selected from the group consisting of a bond and a linear or branched alkyl chain having 1 to 12 carbon atoms, wherein the alkyl chain may be interrupted by one or more C6-C10 aromatic groups and / or by one or more heteroatoms or groups selected from the group consisting of -O-, -S-, -C(O)-, -NR'-, -C(O)NR'-, -NR'-C(O)-, -C(S)NR'-, -NR'-C(S)-NR', and each R' is independently H or a C1-C6 alkyl group.

[0140] In one embodiment, the functionalized statistical chitosan has the following formula (III):

[0141]

[0142] wherein x is 0.25-0.4, typically about 0.3, and y is 0.05-0.2, typically about 0.07.

[0143] In one embodiment, the functionalized statistical chitosan is soluble in aqueous solution at physiological pH (ie, pH 4.8-8) and responds as follows: (DS DOTAGA (%) + 3.5) * (DA (%) + 8) ≥ 150, where DS is the degree of substitution of DOTAGA, and DA is the degree of acetylation of the functionalized statistical chitosan.

[0144] In another embodiment, the functionalized statistical chitosan has formula (III) wherein x is 0.025-0.075, more preferably 0.04-0.06, typically about 0.05, and y is 0.05-0.3, more preferably 0.1-0.2, typically about 0.15.

[0145] In another embodiment, the functionalized statistical chitosan has the following formula (IV):

[0146]

[0147] wherein x is 0.2-0.6, more preferably x is 0.25-0.4, typically about 0.3, and y=z+w is 0.05-0.3, more preferably 0.1-0.2, typically about 0.12.

[0148] Synthesis of functionalized polymers of formula I and II

[0149] The compounds of formula I and II can be synthesized using the methods disclosed in WO2022 / 023677 and the reference Natuzzi, M., Grange, C., Gréa, T. et al. Feasibility study and direct extraction of endogenous freemetallic cations combining hemodialysis and chelating polymer. Sci Rep 11, 19948 (2021).

[0150] Pharmaceutical compositions of functionalized polymers

[0151] The functionalized polymer used may be orally administered per se or may be formulated together with one or more pharmaceutically acceptable excipients.

[0152] Any suitable excipient for use in pharmaceutical compositions known to those of ordinary skill in the art can be used in the compositions described herein.

[0153] In a specific embodiment, the pharmaceutical composition can be a liquid form suitable for oral administration, such as an aqueous solution of the functionalized polymer. In another specific embodiment, the pharmaceutical composition can be a solid dosage form suitable for oral administration. Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In a preferred embodiment, the pharmaceutical composition is a capsule or a tablet.

[0154] In one embodiment, the release of the capsule or tablet contents may be immediate or modified, such as delayed, targeted or extended. In a preferred embodiment, the solid dosage form is an immediate release dosage form.

[0155] A second object of the present disclosure relates to an oral formulation comprising the functionalized polymer of the present disclosure and one or more pharmaceutically acceptable excipients.

[0156] In one embodiment, pharmaceutically acceptable excipients include fillers, disintegrants, lubricants, glidants.

[0157] Fillers

[0158] Fillers (also called dilutants or thinners) are substances added to drug substances to make them suitable for oral administration (e.g., capsules, tablets). Fillers themselves should not produce any pharmacological effects in the human body. Examples of fillers include mannitol, microcrystalline cellulose, lactose monohydrate, anhydrous lactose, corn starch, xylitol, sorbitol, sucrose, dicalcium phosphate, maltodextrin, and gelatin.

[0159] Disintegrants

[0160] Disintegrants are added to oral solid dosage forms to help their depolymerization. Disintegrants are formulated so that the solid dosage form disintegrates rapidly when it comes into contact with moisture. Disintegration is typically considered the first step in the dissolution process. Examples of disintegrants include modified starches, such as sodium starch glycolate, sodium carboxymethyl starch, and pregelatinized starch, cross-linked polymers, such as cross-linked polyvinyl pyrrolidone (crospovidone) or cross-linked sodium carboxymethyl cellulose (croscarmellosesodium) and calcium silicate.

[0161] Lubricants

[0162] Lubricants are substances that we use in tablet and capsule formulations to reduce friction. Lubricants facilitate the extrusion of tablets from the matrix, thereby preventing scratches on its surface. Lubricants can be divided into two groups based on their properties: a) fats and fat-like substances; b) powdered substances. Powdered substances are more suitable than fat-like substances because the latter affect tablet solubility and chemical stability. Powdered lubricants are added by pulverizing the granules. They provide a constant rate of mass outflow for tableting from the hopper to the matrix, thereby ensuring accurate and constant dosage of the drug substance.

[0163] Examples of lubricants include magnesium stearate, hydrogenated castor oil, glyceryl behenate, calcium stearate, zinc stearate, mineral oil, silicone oil, sodium lauryl sulfate, L-leucine, and sodium stearyl fumarate.

[0164] Glidants

[0165] Glidants are mixed with the formulation to enhance the flowability of the tablet-core blend. In the early stages of tableting, glidants are mixed into the particle arrangement of the tablet powder mixture to improve flowability and uniformity within the tablet press die cavity. Glidants promote the flow of tablet particles by reducing the friction of the particles. The effect of glidants on particle flow depends on the size and shape of the particles and the glidant particles. Above a certain concentration, the glidant actually acts to inhibit flowability. In tablet production, glidants are usually added just before compression. Examples of glidants include colloidal silicon dioxide, starch, magnesium stearate, and talc.

[0166] Any suitable excipients for pharmaceutical compositions known to those of ordinary skill in the art may further be used in the compositions described herein.

[0167] The unit dosage of the functionalized polymer is 0.1 mg-500 mg, preferably 1 mg-100 mg, more preferably 2 mg-10 mg, even more preferably about 5 mg.

[0168] Preferred subject populations targeted by the treatment methods

[0169] The treatment methods disclosed herein are suitable for use in subjects suffering from a disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity.

[0170] Without being bound by any theory, it is believed that the present treatment methods disclosed herein prevent the absorption of the heavy metals into the body of the subject.

[0171] In specific embodiments, the subject is a mammal, such as a human subject.

[0172] Thus, the methods of the present disclosure are particularly useful for subjects exposed to heavy metals present in contaminated water and / or food and / or air pollution, for example from contamination, pollution, accidents, war zones such as dirty bombs.

[0173] Examples of diseases associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity include, but are not limited to, kidney disease, liver disease, gastrointestinal disease, cardiovascular disease, respiratory disease, bone disease, brain disease, developmental abnormalities, nervous system and neurobehavioral disorders, diabetes, hearing loss, blood and immune disorders, and cancer disorders.

[0174] The renal disease may in particular be selected from the group consisting of chronic tubulointerstitial nephritis, end-stage renal disease, Fanconi syndrome, chronic kidney disease.

[0175] The liver disease may in particular be selected from the group consisting of non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver fibrosis.

[0176] The gastrointestinal disease may in particular be selected from the group consisting of dysbacteriosis, inflammatory bowel disease such as Crohn's disease or ulcerative colitis.

[0177] The cardiovascular disease may in particular be selected from the group consisting of hypertension, stroke, atherosclerosis, peripheral vascular or arterial disease, coronary artery disease, congestive heart failure.

[0178] The respiratory disease may in particular be selected from the group consisting of pneumonia, pulmonary edema, acute tracheobronchitis, pulmonary fibrosis, asthma, lung cancer.

[0179] The bone disease may in particular be selected from the group consisting of osteomalacia, osteoarthritis, degenerative disc disease and osteoporosis.

[0180] The brain disease may in particular be selected from the group consisting of dementia.

[0181] The developmental abnormality may in particular be selected from the group consisting of congenital and neurological defects, developmental delays and learning disabilities.

[0182] The neurological and neurobehavioral disorders may in particular be selected from the group consisting of autism spectrum disorders.

[0183] The blood and immune disorders may in particular be selected from the group consisting of autoimmune diseases.

[0184] In a specific embodiment, the disease is end-stage renal disease or inflammatory bowel disease.

[0185] In a specific embodiment, the disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity is due to the absorption of radioactive isotopes of heavy metals as described herein in contaminated areas after nuclear accidents or terrorism or war, thereby polluting and contaminating water and / or air and / or food.

[0186] Treatment

[0187] Functionalized polymers as disclosed in part above, and more preferably functionalized statistical chitosan, and pharmaceutical compositions thereof, can be used as medicaments in a method of treating a disease in a subject in need thereof, and more particularly in a subpopulation of subjects as defined above, which disease is associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity.

[0188] The functionalized polymer is orally administered to a subject in an amount sufficient to prevent absorption of the heavy metal in the subject.

[0189] The pharmaceutical compositions may be administered orally in any manner determined by one of ordinary skill in the medical arts to be appropriate for the disease or condition to be treated.

[0190] The appropriate dosage and the appropriate duration and frequency of administration will be determined by factors as discussed herein, including the condition of the subject, the type and severity of the subject's disease, the particular form of the active ingredient and the method of administration.

[0191] In general, an appropriate dose (or effective dose) and treatment regimen provides the pharmaceutical composition in an amount sufficient to achieve a therapeutic effect, e.g., an improved clinical outcome, such as more complete or partial remissions, or longer disease-free and / or overall survival, or a reduction in symptom severity or other benefits described in detail herein.

[0192] In one embodiment, the functionalized polymer is administered to a subject in need thereof once, twice, or three times per day.

[0193] A typical daily dosage may comprise 0.1 mg-500 mg, preferably 1 mg-100 mg, more preferably 2 mg-10 mg, even more preferably about 5 mg, preferably of the functionalized statistical chitosan. BRIEF DESCRIPTION OF THE DRAWINGS

[0194] Figure 1: Fluorescence qualitative biodistribution in kidney, liver, brain, spleen, heart, urine, lung, bone, skin, blood, muscle, stomach, intestine and colon after 1, 2, 4 and 24 hours (Example 1).

[0195] Figure 2 : Average percentage of administered Gd within the digestive tract over 24 hours (Example 1).

[0196] Figure 3 : Body weight changes over 14 days in mice administered MEX-CD1 daily (Example 2).

[0197] Figure 4 : Hematology after 14 days of daily administration of MEX-CD1 in mice (Example 2). Hematology was determined after the mice were sacrificed. These results provide evidence of acute heavy metal poisoning in saline-treated mice. All relationships are not significant except for the following definitions: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0198] Figure 5 : Blood concentrations of lead and cadmium after 14 days of daily administration of MEX-CD1 in mice (Example 2). All mice were sacrificed after 15 days of exposure. NS p>0.05, *p<0.05, **p<0.01, ***p<0.001.

[0199] Figure 6 : Graphical representation of the chelating capacity of MEX-CD1 for different concentrations of lead and cadmium (Example 4). Technique used: HPLC-MS.

[0200] Figure 6a : HPLC-MS chromatogram detecting the interaction between lead and MEX-CD1 (Example 4).

[0201] Figure 6b : HPLC-MS chromatogram detecting the interaction between cadmium and MEX-CD1 (Example 4).

[0202] Figure 7 : High performance liquid chromatography-mass spectrometry (HPLC-MS) was used to directly prove that MEX-CD1 has lead chelating effect (Example 4).

[0203] Figure 8 : Graphical representation of the lead chelating capacity of MEX-CD1 (Example 4). Technique used: ICP-MS.

[0204] Fig. 9 : Graphical representation of the chelating capacity of MEX-CD1 for lead and cadmium (Example 4). Technique used: ICP-MS. DETAILED DESCRIPTION

[0205] The present disclosure is described in more detail and more specifically below in conjunction with embodiments, but these embodiments are not intended to limit the present invention.

[0206] Materials and methods:

[0207] Solution preparation:

[0208] Solution 1: 10g / L MEX-CD1

[0209] Lyophilized MEX-CD1 was dissolved in ultrapure water at a concentration of 10 g / L.

[0210] Solution 2: 10 g / L MEX-DTPA

[0211] Lyophilized MEX-DTPA was dissolved in ultrapure water at a concentration of 10 g / L.

[0212] Solution 3: 10 g / L MEX-DOTAM

[0213] Lyophilized MEX-DOTAM was dissolved in ultrapure water at a concentration of 10 g / L.

[0214] Solution 4: Biodistribution of MEX-CD1 mixture

[0215] The lyophilized MEX-CD1 obtained by the synthesis method described below was further labeled with gadolinium and dissolved in ultrapure water at a concentration of 10 g / L. The lyophilized MEX-CD1 labeled with Cyanine 5.5 was dissolved in ultrapure water at a concentration of 10 g / L. 1 mL of each solution was combined to obtain a 5 g / L mixture solution of each labeled MEX-CD1.

[0216] Solution 6: 5g / L MEX-CD1 + 3.5g / L NaCl

[0217] 3.5 g / L NaCl was dissolved in ultrapure water. Lyophilized MEX-CD1 was then dissolved in the solution to a final concentration of 5 g / L. The solution was sterilized at 121°C for 20 minutes.

[0218] Solution A: Concentrated saline solution for in vitro laboratory testing

[0219] 263g / L NaCl, 3.35g / L KCl, 6.24g / L CaCl2, 2.14g / L MgCl2, 10.8g / L acetic acid, 45g / L glucose in ultrapure water

[0220] Solution B: 0.1 M acetate buffer, pH 4.6

[0221] Acetate buffer, containing 11.4 mL acetic acid (MS grade), 15.4 g ammonium acetate and 2 L ultrapure water.

[0222] Experimental protocol:

[0223] Solution 1 :High pressure liquid chromatography-mass spectrometry (HPLC-MS)

[0224] The eluent used in this method was acetate buffer (solution B, prepared with MS-grade qualified products), the injection volume was 10uL, and the flow rate was fixed at 0.4mL / min. The chromatographic column used was SEC Polysep GFC-P 4000 series. HPLC was coupled to an inductively coupled plasma mass spectrometer (ICP-MS) for detection of the selected isotopes.

[0225] Solution 2 :Vivaspin ultrafiltration experiment

[0226] A 20 ml Vivaspin centrifuge tube was used, which has a 30 kDa inner membrane. Centrifugation was performed at a speed of 4000 rpm, with each cycle lasting 10-30 minutes. After each centrifugation, for each sample, the solution that passed through the membrane was collected, which was later referred to as the lower supernatant. This operation was performed once, and ideally, about 0.5 mL of the supernatant remained and was collected for ICP-MS analysis.

[0227] The samples (including the original solution, the supernatant and the lower supernatant) were analyzed by ICP-MS to determine the concentration of the target metal in each solution. The samples were diluted (at least) 10 times using 1% HNO3. An internal standard (indium) was added to each sample to a final concentration of 2ppb In. The analysis was performed in KED (Kinetic Energy Discrimination) mode or standard mode, where helium is introduced into the chamber. KED was used for analysis, which was combined with analysis of other more sensitive elements, but most samples were performed in standard mode: 208Pb, 206Pb, 111Cd, 112Cd, 114Cd.

[0228] Calibration curves ranging from 0.01-10 ppb were prepared for lead and cadmium so that the system would convert the counts per second recorded for each sample into concentrations in ppb.

[0229] Synthesis of functionalized polymers of the present invention:

[0230] MEX-CD1 (chitosan as polymer and DOTAGA as chelating moiety)

[0231] In the first step, 60 g of chitosan, 4 L of ultrapure water and 45 mL of glacial acetic acid were added to a 10 L reactor and stirred at a pH of 4.5 ± 0.5 for 16 h. A light yellow solution was observed.

[0232] In the second step, 1.2 L of 1,2-propylene glycol were added to the light yellow solution obtained from step 1 and stirred for 1 hour. A solution consisting of 14 mL of acetic anhydride and 600 mL of 1,2-propylene glycol was added slowly over 10 minutes to obtain uniform acetylation along the polymer chains. The medium was kept and stirred for 4 hours.

[0233] The acetylation rate can be determined by elemental analysis. The molar mass of the non-acetylated monomer (monomer B) of the polymer is 161.2 gmol -1 (C6NO4H 11 ), and the molar mass of the acetylated monomer (monomer A) of the polymer is 203.2 gmol -1 (C8NO5H 13 ). The elemental analysis of the polysaccharide obtained after step 2 was as follows: C 39.22%; H 7.55%; and N 6.77%, which corresponds to an acetylation rate of 29% (x=0.29), as determined by 1H NMR.

[0234] In the third step, 2L of the solution obtained after the second step was stirred. 120g of DOTAGA anhydride was added and stirred for 16 hours. At the end of the reaction, the solution was diluted 10 times in ultrapure water and purified by tangential purification using a 100kDa membrane. After the first dilution, the final volume was 16L, and the solution was filtered with 480L of 0.1M acetic acid, maintaining a constant volume of 16L, and then filtered with 320L of ultrapure water. The volume was 8L when the purification was re-concentrated. HPLC-UV can verify that unreacted DOTAGA has been successfully removed from the solution. Then, before freeze-drying, a solution with a concentration of 10g / L was filtered with a nylon filter (0.4μm).

[0235] 1 H NMR can determine the percentage of monomer functionalized with DOTAGA(y) relative to a polymer of known acetylation percentage x.

[0236] The amount of grafted DOTAGA was determined by spectrophotometric UV dosimetry at 295 nm. MEX-CD1 contained 0.345 mmol DOTAGA / g polymer.

[0237] The mole fraction of N-acetylglucosamine repeating unit x was determined to be x=0.29 using 1H-NMR, the mole fraction of DOTAGA grafted repeating unit y was determined to be y=0.075 using 1H-NMR, and the dosage of copper was determined using HLPC-SEC.

[0238] MEX-DTPA (chitosan is the polymer and DTPA is the chelating moiety)

[0239] In the first step, 1 g of chitosan, 66 mL of ultrapure water and 0.84 mL of glacial acetic acid were added and stirred at a pH of 4.5 ± 0.5 for 16 h. A light yellow solution was observed.

[0240] In the second step, 20 mL of 1,2-propylene glycol was added to the light yellow solution obtained from step 1 and stirred for 1 hour. A solution consisting of 0.704 mL of acetic anhydride in 30 mL of 1,2-propylene glycol was added slowly over 10 minutes to obtain uniform acetylation along the polymer chains. The solution was kept and stirred for 4 hours.

[0241] In the third step, 0.8 g of DTPA dianhydride was added to the previous solution and stirred for 16 hours. At the end of this reaction, the solution was diluted 10 times in ultrapure water and purified by tangential purification using a 100 kDa membrane. After the first dilution, the final volume was 1 L, and the solution was filtered with 5 L of 0.1 M acetic acid, keeping the volume constant at 1 L, and then filtered with 5 L of ultrapure water. The volume at the end of the purification was re-concentrated to ~100 mL. HPLC-UV can verify that the unreacted DTPA has been successfully removed from the solution.

[0242] The amount of grafted DTPA was determined using spectrophotometric UV dose at 295 nm. MEX-DTPA contained 0.21 mmol DTPA / g polymer.

[0243] The molar fraction of the N-acetylglucosamine repeating unit x was determined to be x=0.58 using 1H-NMR, and the molar fraction of the DTPA-grafted repeating unit y was determined to be y=0.045 using 1H-NMR.

[0244] The MEX-DTPA synthesized herein can be used in the therapeutic methods described in the present disclosure.

[0245] MEX-DOTAM (chitosan is the polymer and DOTAM is the chelating moiety)

[0246] In the first step, 1 g of chitosan, 66 mL of ultrapure water and 0.84 mL of glacial acetic acid were added and stirred at a pH of 4.5 ± 0.5 for 16 h. A light yellow solution was observed.

[0247] In the second step, 20 mL of 1,2-propylene glycol was added to the light yellow solution obtained from step 1 and stirred for 1 hour. A solution consisting of 0.704 mL of acetic anhydride in 30 mL of 1,2-propylene glycol was slowly added over 10 minutes to obtain uniform acetylation along the polymer chain. The medium was kept and stirred for 4 hours. The solution was then purified by tangential purification using a 100 kDa membrane and 7 L of ultrapure water to remove the solvent and re-concentrate to about 100 mL. The pH of the solution was raised to 7 ± 0.1 using NaOH. 8 ml of DMSO was added to the solution to form a 5% DMSO aqueous solution environment and stirred for 1 hour.

[0248] In the third step, 11 mL of a 100 g / L solution of DOTAM NHS-ester in DMSO was added over 10 minutes. The solution was then diluted 2-fold with ultrapure water and stirred for 16 hours. At the end of the reaction, the solution was diluted 10-fold in ultrapure water and purified by tangential purification using a 100 kDa membrane. After the first step of dilution, the final volume was 1 L, and the solution was filtered with 5 L of 0.1 M acetic acid, keeping the volume constant at 1 L, and then filtered with 5 L of ultrapure water. The volume was about 100 mL when the purification was re-concentrated. HPLC-UV can verify that unreacted DOTAM has been successfully removed from the solution.

[0249] The amount of grafted DOTAM was determined using spectrophotometric UV dose at 295 nm. The MEX-DOTAM contained 0.049 mmol DOTAM / g polymer.

[0250] The molar fraction of the N-acetylglucosamine repeating unit x was determined to be x=0.58 using 1H-NMR, and the molar fraction of the DOTAGA-grafted repeating unit y was determined to be y=0.0097 using 1H-NMR.

[0251] The MEX-DOTAM synthesized herein can be used in the therapeutic methods described in the present disclosure.

[0252] PEG-DTPA (PEG is the polymer and DTPA is the chelating moiety)

[0253] The PEG used was an eight-arm PEG amine with a molecular weight of 40 kDa and having the following formula (V):

[0254]

[0255] Where n is 111.3, purchased from Creative PEGWork. DTPA dianhydride was provided by Chematech. Synthesis was performed in DMSO (Fischer Chemicals). The VIVAFLOW 30 kDa kit used for purification was purchased from Sartorius.

[0256] 0.36 g of DTPA dianhydride was dissolved in 10.26 mL of DMSO (35 g / L) and stirred at 40°C for 1 hour. At the same time, 0.27 g of PEG was weighed and dissolved in 43.36 mL of DMSO. Then, the PEG solution was directly added to the DTPA solution and stirred at 40°C for 1.5 hours. Ultrapure water and 10 -2 M HCl and VIVAFLOW 30 kDa cassette purification solution, final purification factor ~10,000.

[0257] The amount of grafted DTPA was determined using a spectrophotometric UV dose at 295 nm. PEG-DTPA contained 0.18 mmol DTPA / g polymer. Using 1H-NMR, the molar fraction of the repeating unit y grafted with DTPA was y=1.

[0258] The structure of the synthesized PEG-DTPA has the following formula (VI):

[0259]

[0260] Where n is 111.3.

[0261] The PEG-DTPA synthesized herein can be used in the therapeutic methods described in the present disclosure.

[0262] PEG-DOTAM (PEG is the polymer and DOTAM is the chelating moiety)

[0263] The PEG used was an eight-arm PEG amine with a molecular weight of 40 kDa, purchased from Creative PEGWork. DOTAM NHS-ester was provided by Chematech as a 100 g / L solution in DMSO. Synthesis was performed in DMSO (Fischer Chemicals). The kit VIVAFLOW 30 kDa used for purification was purchased from Sartorius.

[0264] 3.5 mL of 100 g / DOTAM NHS-ester / DMSO solution was added to a round-bottom flask and stirred at 40 ° C for 1 hour. At the same time, 0.27 g of PEG was weighed and dissolved in 43.36 mL of DMSO. Then, the PEG solution was directly added to the DTPA solution and stirred at 40 ° C for 1.5 hours. Ultrapure water and 0.1 M acetic acid and VIVAFLOW 30 kDa box were used to purify the solution, and the final purification factor was about 10,000.

[0265] The amount of grafted DOTAM was determined using a spectrophotometric UV dose at 295 nm. The PEG-DOTAM contained 0.036 mmol DOTAM / g polymer. Using 1H-NMR, the molar fraction of DOTAM grafted on the amine functional groups of the ethylene glycol monomer y was y=0.21.

[0266] The structure of the synthesized PEG-DOTAM has the following formula (VII):

[0267]

[0268] Where n is 111.3.

[0269] The PEG-DOTAM synthesized herein can be used in the therapeutic methods described in the present disclosure.

[0270] Example 1: Biodistribution of Orally Administered MEX-CD1 in Mice

[0271] 8 mice were orally administered 0.2 ml of solution 4. 2 mice were sacrificed at each time point: 1, 2, 4 and 24 hours. Fluorescence studies were performed with a CCD camera and the following parameters: 2 sets of spotlights at 633 and 470 nm excitation, and 2 sets of filters at 680 ± 20 and 520 ± 20 nm excitation, and then the organs were digested in nitric acid for gadolinium quantification using ICPMS.

[0272] result

[0273] Fluorescence qualitative biodistribution

[0274] like Figure 1 As shown, no fluorescence was observed at any time point in the kidney, liver, brain, spleen, heart, urine, lung, bone, skin, blood, and muscle after 1, 2, 4, and 24 hours. Fluorescence was only observed in the stomach, intestine, and colon after 1, 2, and 4 hours. A small amount of autofluorescence was observed in the stomach of control mice.

[0275] Fluorescence results showed that MEX-CD1 was only present in the stomach, intestines and colon (digestive tract). Some autofluorescence was observed in the stomach of control mice, which explained the remaining fluorescence in both stomachs after 24 hours.

[0276] Quantitative biodistribution by ICP-MS

[0277] like Figure 2 As shown, there were two experimental mice at each time point, so the average gadolinium percentage was taken. Figure 1 The trend shows that within the first hour, most of the MEX-CD1 is already present in the intestine, and within 24 hours, all of the MEX-CD1 is excreted from the digestive system of the mice.

[0278] Other organs were also evaluated, and gadolinium concentrations in all organs were less than 1% of the administered gadolinium and were therefore considered zero.

[0279] Therefore, orally administered MEX-CD1 remained in the digestive tract, did not pass through the intestinal membrane into the blood, and was completely eliminated after one day.

[0280] Example 2: Oral administration of MEX-CD1 to mice exposed to heavy metals

[0281] The study was conducted as follows:

[0282] Control group - fed normal rodent chow and water, no treatment

[0283] Saline group - fed contaminated rodent chow, normal water, and oral administration of saline solution once daily for 14 days

[0284] MEX-CD1 group - fed contaminated rodent chow, normal water, and orally administered MEX-CD1 solution once daily for 14 days

[0285] Contaminated rodent feed = 7ppm (mg / kg) Cd + 50ppm (mg / kg) Pb

[0286] Oral administration = 0.2 mL 3.5 NaCl / H2O or 0.2 mL 5 g / L MEX-CD1 + 3.5 NaCl / H2O

[0287] result

[0288] weight

[0289] like Figure 3 As shown, the body weight reduction observed in the saline and MEX-CD1 treated groups can be attributed to exposure to contaminated rodent chow. Body weights of the groups stabilized after 6 days. This demonstrates that oral administration of MEX-CD1 did not affect the health of the mice, as evidenced by changes in body weight over time.

[0290] Liver enzyme activity

[0291] Table 1: Liver enzyme activities in mice after daily oral administration of saline or MEX-CD1 for 14 days

[0292]

[0293] The enzyme activity levels of mice in all groups were not statistically significant, demonstrating that oral administration of MEX-CD1 did not affect liver function.

[0294] hematology

[0295] like Figure 4As shown, some hematological parameters in saline-treated mice (particularly white blood cells (WBC) and MCHC, which showed significant differences, and platelets (PLT) and MCH with slight changes) indicated the presence of acute heavy metal poisoning. This acute heavy metal poisoning occurred in a manner similar to observations previously described in the literature. After oral administration of MEX-CD1, the degree of poisoning returned to normal levels. This not only proves that this treatment has no negative impact on hematological parameters, but also indicates the beneficial effect of this oral treatment in preventing exposure to heavy metals.

[0296] Lead and cadmium blood levels

[0297] like Figure 5 As shown, when mice were exposed to contaminated rodent feed, blood concentrations of lead and cadmium increased significantly. For lead, MEX-CD1 helped mitigate blood concentrations in mice by once-daily oral administration. Since MEX-CD1 was present at trace levels, it had little effect on cadmium concentrations.

[0298] Example 3: In vitro chelation of lead and cadmium by MEX polymers in a complex environment

[0299] Polymer 1-MEX-CD1

[0300] Samples were prepared using Solution A as defined in the Materials and Methods section. Metal standards purchased from SCP Science were used, with a concentration of 1000 ppm Pb in 5% HNO3. 2+ or Cd 2+ , and solution 1, prepare 4 samples. Use 1M NaOH to adjust pH. According to scheme 2, the samples are experimentally analyzed.

[0301] Table 2

[0302]

[0303] Table 3

[0304]

[0305] Polymer 2-MEX-DTPA

[0306] Samples were prepared using Solution A. Metal standards purchased from SCP Science were used, with a concentration of 1000 ppm Pb in 5% HNO3. 2+ or Cd 2+ , and solution 2, prepare 7 samples. Adjust pH with 1M NaOH. Perform experimental analysis on the samples according to Scheme 2.

[0307] Table 4

[0308] sample Alkali solution used MEX-DTPA concentration Pb and Cd concentration (ppb) pH 1.1 Solution A 0 100 2 1.2 Solution A 0 50 2 2 Solution A 0.1g / L 100 2 3 Solution A 1g / L 50 2 4 Solution A 1g / L 50 3 5 Solution A 1g / L 50 4 6 Solution A 1g / L 50 5

[0309] Table 5

[0310]

[0311] Polymer 3-MEX-DOTAM

[0312] Samples were prepared using Solution A. Metal standards purchased from SCP Science were used with a concentration of 1000 ppm Pb in 5% HNO3 2+ or Cd 2+ , and solution 3, prepare 7 samples. Use 1M NaOH to adjust pH. According to scheme 2, the samples are experimentally analyzed.

[0313] Table 6

[0314]

[0315] Table 7

[0316]

[0317] Since the original solution was centrifuged and passed through the membrane once, the original concentration and supernatant concentration were comparable. For each polymer, effective chelation of cadmium and lead was observed at pH 4 and 5, as indicated by lower metal concentrations in the supernatant compared to the original concentration. Minimal lead chelation was observed for MEX-DOTAM and MEX-DTPA at pH 2 and 3, while effective chelation was observed at pH 4 and 5. Since MEX-DTPA and MEX-DOTAM have more effective chelation abilities in low pH environments compared to MEX-CD1, these polymers may show similar or improved chelation abilities to MEX-CD1 in vivo.

[0318] Example 4: MEX-CD1 Chelates Lead and Cadmium in Simple and Complex Environments in Vitro MEX-CD1 Chelates Lead and Cadmium in Water

[0319] Using a Pb and Cd metal mixture purchased from SCP Science at a concentration of 1000 ppm in 5% HNO3, and Solution 1, the following 6 samples were prepared in ultrapure water and analyzed by HPLC-MS according to Protocol 1.

[0320] Table 8

[0321]

[0322] Figure 6 , 6a and 6b show that the polymer can effectively chelate lead at a concentration of 0.1 g / L in a concentrated solution composed of salt and acid. Figure 6a and6b A larger polymer peak was observed at 11 min, corresponding to a higher degree of chelation of these metals on the polymer. Figure 6 A linear relationship can be seen in the Figure 3, which demonstrates the chelation of these metals by MEX-CD1 within the desired concentration range.

[0323] MEX-CD1 chelation in solution A

[0324] Add 77 ppb of Pb to solution A 2+ This doping solution, as well as solution 1, solution B, and ultrapure water, were used to generate different acidic environments. Chelation was tested in the following different environments: concentrated acid (pH 2), pH 5, and pH 7.4. The pH was adjusted with NaOH, and the samples were analyzed by HPLC-MS according to protocol 1.

[0325] Table 9

[0326] sample Dilution factor of solution A pH Concentrated solution A 1 2 Concentrated solution A + 0.1g / L MEX-CD1 1 2 Solution A + 0.1 g / L MEX-CD1 5 5 Solution A + 0.1 g / L MEX-CD1 5 7.4

[0327] above Figure 7 It was shown that the polymer can effectively chelate lead at a concentration of 0.1 g / L in a concentrated solution composed of salt and acid. At pH 5 and 7.4, most of the lead was chelated and coupled to the polymer, which was indicated by the chromatogram of HPLC / SEC-ICP-MS (lead detection) with an increase in peak area at 15 minutes, specifically corresponding to the polymer.

[0328] MEX-CD1 in water

[0329] Ultrapure water, solution 1 and 50000ppm Pb 2+ The samples were prepared in 5% HNO3 solution. According to protocol 2, the samples were subjected to experimental analysis.

[0330] Table 10

[0331]

[0332] above Figure 8 Shows a linear relationship between increasing lead concentration and MEX-CD1 chelation. MEX-CD1 in water, ultra-trace chelation

[0333] A metal mixture solution containing 10 ppm of Pb and Cd in 5% HNO3 (purchased from SCP Science) was used to prepare the samples along with Solution 1 and ultrapure water. Protocol 2 was used to experimentally analyze these samples.

[0334] Table 11

[0335]

[0336] Fig. 9The results show that ICP-MS analysis shows that the chelation efficiency of lead and cadmium starts from 0.05 ppb (50 ppt). Even in the range of 0.01 ppb (10 ppt), lead shows effective chelation.

Claims

1. A functionalized polymer for use in treating a disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity in a subject in need thereof, wherein the functionalized polymer is soluble in aqueous solution, has a weight average molecular weight of 30 kDa to 5000 kDa, and wherein a portion of the monomer units are functionalized with a chelating moiety, wherein a therapeutically effective amount of the functionalized polymer is orally administered to the subject.

2. The functionalized polymer for use thereof according to claim 1, wherein the subject in need is a subject exposed to heavy metals present in contaminated water and / or food and / or air pollution.

3. The functionalized polymer for use thereof according to claim 1 or 2, wherein the functionalized polymer comprises at least 1 w% of the chelating moiety, such as 1 w%-40 w%.

4. A functionalized polymer for use according to any one of claims 1 to 3, wherein the chelating moiety is selected from the group consisting of DOTA, NOTA, NODAGA, DOTAGA, DOTAM, NOTAM, DOTP, NOTP, TETA, TETAM, DTPA, Bz-DFO, DFO and mixtures thereof, preferably selected from the group consisting of DOTAGA, Bz-DFO, DFO, DOTAM, DTPA and mixtures thereof.

5. The functionalized polymer for use thereof according to any one of claims 1 to 4, wherein the disease associated with heavy metal-induced toxicity or heavy metal-induced carcinogenicity is selected from the group consisting of kidney disease, liver disease, gastrointestinal disease, cardiovascular disease, respiratory disease, bone disease, brain disease, developmental abnormalities, nervous system and neurobehavioral disorders, diabetes, hearing loss, blood and immune diseases, and cancer diseases.

6. The functionalized polymer for use according to claim 5, wherein: a. The kidney disease is selected from the group consisting of chronic tubulointerstitial nephritis, end-stage renal disease, Fanconi syndrome, chronic kidney disease; b. The liver disease is selected from the group consisting of non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), and liver fibrosis; c. The gastrointestinal disease is selected from the group consisting of dysbacteriosis, inflammatory bowel disease such as Crohn's disease or ulcerative colitis; d. The cardiovascular disease is selected from the group consisting of hypertension, stroke, atherosclerosis, Peripheral vascular or arterial disease, coronary artery disease, congestive heart failure; e. The respiratory disease is selected from the group consisting of pneumonia, pulmonary edema, acute tracheobronchitis, pulmonary fibrosis, asthma, lung cancer; f. The bone disease is selected from the group consisting of osteomalacia, osteoarthritis, degenerative disc disease and osteoporosis; g. The brain disease is selected from the group consisting of: dementia; h. The developmental abnormality is selected from the group consisting of congenital and neurological defects, developmental delays and learning disabilities; i. The nervous system and neurobehavioral disorders are selected from the group consisting of autism spectrum disorders; or, j. The blood and immune diseases are selected from the group consisting of autoimmune diseases.

7. A functionalized polymer for use according to any one of claims 1 to 6, wherein the heavy metal is selected from the group consisting of lead (Pb), cadmium (Cd), arsenic (As), chromium (Cr), mercury (Hg), cobalt (Co), aluminum (Al), antimony (Sb), barium (Ba), bismuth (Bi), gallium (Ga), germanium (Ge), gold (Au), indium (In), nickel (Ni), platinum (Pt), silver (Ag), strontium (Sr), tellurium (Te), thallium (Tl), tin (Sn), titanium (Ti), vanadium (V), plutonium (Pu), cesium (Cs), cerium (Ce), zirconium (Zr), ruthenium (Ru), technetium (Tc), radium (Ra), thorium (Th), americium (Am), iridium (Ir), californium (Cf), polonium (Po) and uranium (U), or a mixture thereof, preferably selected from the group consisting of lead, cadmium, arsenic, chromium and mercury, more preferably lead or cadmium.

8. The functionalized polymer for use thereof according to any one of claims 1 to 7, wherein the functionalized polymer is administered to the subject in a unit dose of 0.1 mg-500 mg, preferably 1 mg-100 mg, more preferably 2 mg-10 mg, even more preferably about 5 mg.

9. A functionalized polymer for use thereof according to any one of claims 1 to 8, wherein the functionalized polymer is selected from the group consisting of polysaccharides such as chitosan, polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polyallylamine (PAH), wherein a portion of the monomer units are functionalized with a chelating moiety.

10. The functionalized polymer for use according to any one of claims 1 to 9, wherein the functionalized polymer is a functionalized statistical chitosan having a weight average molecular weight of 100 kDa to 1000 kDa and having formula (I): in Each Rc is a chelating moiety, Each Z is independently a linker which is a single bond or a hydrocarbon chain containing 1 to 12 carbon atoms, the hydrocarbon chain being linear or branched and optionally containing one or more unsaturated groups and one or more heteroatoms, the heteroatoms being preferably selected from nitrogen, oxygen, sulfur and halogens, x is 0.005-0.7, preferably 0.05-0.7, y is 0.005-0.5, preferably 0.01-0.2, The ratio y / x is greater than or equal to 0.01, preferably greater than or equal to 0.02, and The sum of x+y is greater than or equal to 0.15, preferably greater than or equal to 0.

30.

11. The functionalized polymer for use thereof according to claim 10, which has the following formula (III): wherein x is 0.25-0.4, typically about 0.3, and y is 0.05-0.2, typically about 0.

07.

12. The functionalized polymer for use thereof according to any one of claims 1 to 11, wherein the functionalized polymer is formulated into a solid dosage form.

13. The functionalized polymer for use thereof according to any one of claims 1 to 12, wherein the functionalized polymer is administered once, twice or three times a day.

14. An oral formulation comprising the functionalized polymer according to any one of claims 1 to 13 and one or more pharmaceutically acceptable excipients, wherein the unit dose of the functionalized polymer is 0.1 mg-500 mg, preferably 1 mg-100 mg, more preferably 2 mg-10 mg, even more preferably about 5 mg.

15. The oral formulation according to claim 14, which has the following formula (III): wherein x is 0.25-0.4, typically about 0.3, and y is 0.05-0.2, typically about 0.07.

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