A macromolecular heavy metal complexing agent containing 8-hydroxyquinoline and amido phosphate structure and a preparation method thereof

Abstract

The present application relates to a kind of macromolecular heavy metal complexing agent containing 8-hydroxyquinoline and amido phosphate structure and its preparation method, the present application also relates to a kind of heavy metal antidote, and the use of the macromolecular heavy metal complexing agent of the present application in preparing drug for treating heavy metal poisoning.The macromolecular complexing agent of the present application has excellent binding capacity with cadmium, lead, mercury and other heavy metal ions, and is high in safety, has excellent and long-acting heavy metal protection effect.The preparation method of the present application can mass, simply and efficiently prepare the macromolecular complexing agent of the present application, and preparation process is simple, high in yield, raw material is cheap and easy to obtain, reaction condition is mild.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of new chemical materials, and particularly relates to a macromolecular heavy metal complexing agent containing 8-hydroxyquinoline and amine phosphonate structure. BACKGROUND

[0002] Cadmium, lead, mercury and other heavy metals are important pollutants in the environment, which are extremely toxic and have characteristics of long-term, hysteresis, high risk and the like, and are difficult to be degraded through physical, chemical and biological approaches. Heavy metals mainly enter the human body through respiratory tract, digestive tract and skin, and can be long-term accumulated in the body. A large number of studies have confirmed that heavy metals can act on various proteins and enzymes in the human body, affecting their physiological functions; at the same time, heavy metals in the environment can induce the body to produce excessive active oxygen, stimulate the body to occur oxidative stress phenomenon, and then induce cell and body damage. Therefore, it has very important research significance and application value to develop safe and efficient heavy metal antidotes to rescue heavy metal poisoning population.

[0003] At present, most of the heavy metal antidotes used in clinical research are small molecule compounds, such as dimercaprol, sodium dimercaptosuccinate, ethylenediaminetetraacetic acid and the like, which can effectively complex heavy metals.

[0004] In the field of heavy metal adsorption materials, there are reports of immobilizing small molecule ligands on the surface of resins. For example, patent document 1 discloses a solidified 8-hydroxyquinoline type chelating adsorption material, which is obtained by bonding the ligand 8-hydroxyquinoline to the surface of silica gel microparticles of polyhydroxyethyl methacrylate. The chelating adsorption material can be used for removing metal ion pollution and trace enrichment analysis in water bodies.

[0005] CITATIONS

[0006] Patent document 1: CN102773082A. SUMMARY

[0007] Problems to be solved by the application

[0008] The small molecule heavy metal antidotes in the prior art all have defects of large toxicity, fast in vivo metabolism and the like, and therefore have limited use effect in the clinic. Although the chelating adsorption material obtained by immobilizing small molecule ligands on the surface of resins in patent document 1 can be used for water purification, it cannot be dissolved or dispersed, and cannot be used as a heavy metal antidote.

[0009] Therefore, it is urgent to develop a safe, effective and clinically applicable heavy metal complexing agent.

[0010] Solution to the problem

[0011] In view of the above problems of heavy metal antidotes in the prior art, the present inventors have developed a macromolecular heavy metal complexing agent containing 8-hydroxyquinoline and amine phosphate ester structures and a preparation method thereof.

[0012] Specifically, the present application solves the problems of the present application by the following solutions.

[0013] [1] A polymer, characterized by containing a structural unit I represented by Formula I and a structural unit II represented by Formula II:

[0014]

[0015] wherein R1 and R2 each independently represent an alkyl group having 1 to 10 carbon atoms or a phenyl group optionally substituted with an alkyl group, R3 and R4 each independently represent hydrogen or a methyl group, and Z represents a single bond, -O-, -S-, or -NR 0 - wherein R 0 is hydrogen or an alkyl group having 1 to 5 carbon atoms, B represents a single bond or an alkylene group having 1 to 10 carbon atoms, X represents a single bond, -O-, -S-, -CO-, -NR 0 -, -C(=O)NH-, -C(=O)O-, -OC(=O)-, or an alkylene group having 1 to 10 carbon atoms, and G represents a hydrophilic group or a hydrophilic segment.

[0016] [2] The polymer according to [1], characterized in that the weight average molecular weight thereof is 1000 to 500000; and the molar ratio of the structural unit I to the structural unit II is (0.5 to 1.5):(0.5 to 10).

[0017] [3] The polymer according to [1] or [2], characterized in that, in Formula I, Z represents -O- or -NR 0 -, B represents an alkylene group having 1 to 10 carbon atoms; and in Formula II, G represents a hydroxyl group, a carboxyl group, a sulfonic acid group, a phosphoric acid group, a sulfuric acid group, an amide group, an amine group, a quaternary ammonium salt group, a morpholino group, or a polyethylene glycol segment, and X represents a single bond, -CO-, -C(=O)NH-, or -C(=O)O-.

[0018] [4] The polymer according to [3], characterized in that, in Formula I, R1 and R2 each independently represent a methyl group, an ethyl group, an n-butyl group, an iso-butyl group, a phenyl group, or a benzyl group, Z represents -NH-, and B represents an alkylene group having 1 to 5 carbon atoms; and in Formula II, X represents -CO-, -CONH-, or -COO-; and G is a polyethylene glycol segment, an amine group, or a morpholino group.

[0019] [5] A method for producing the polymer according to any one of [1] to [4], characterized by comprising the following steps:

[0020] Step 1: reacting compound 1 or its hydrochloride, compound 2 and compound 3 in the presence of an acid-binding agent to obtain monomer 1;

[0021]

[0022] Step 2: polymerizing monomer 1 and monomer 2 in the presence of a free radical polymerization initiator;

[0023]

[0024] wherein each group is defined in any one of [1] to [4].

[0025] [6] The preparation method according to [5], characterized in that the acid-binding agent is triethylamine, pyridine or 4-dimethylaminopyridine; and the free radical initiator is one or more selected from azo initiators and peroxide initiators.

[0026] [7] The preparation method according to [5] or [6], characterized in that in step 1, the molar ratio of compound 1 or its hydrochloride, the acid-binding agent, compound 3 and compound 2 is (0.8-1.2):(0.8-1.2):(0.9-1.3):(1.0-1.4), preferably 1:1:1.1:1.2; and in step 2, the molar ratio of the free radical initiator and monomer 1 is (0.01-0.1):1, and the molar ratio of monomer 1 and monomer 2 is (0.5-1.5):(0.5-10).

[0027] [8] The preparation method according to [5] or [6], characterized in that the reaction temperature of step 1 is 35-80℃, and the reaction time is 2-12h; and the polymerization reaction of step 2 is carried out under anaerobic conditions, the polymerization temperature is 50-100℃, and the polymerization time is 4-24h.

[0028] [9] A heavy metal antidote, characterized in that it comprises the polymer according to any one of [1] to [4] and a pharmaceutically acceptable carrier.

[0029]

[10] Use of the polymer according to any one of [1] to [4] in the preparation of a medicament for treating heavy metal poisoning.

[0030] Effects of the invention

[0031] The macromolecular complexing agent of the present application has excellent binding capacity for heavy metal ions such as cadmium, lead and mercury, is highly safe, and has excellent and long-acting heavy metal protection effect. The preparation method of the present application can mass-produce, simply and efficiently produce the macromolecular complexing agent of the present application, and has the advantages of simple preparation process, high yield, cheap and readily available raw materials, and mild reaction conditions. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 is the synthetic route map of Example 1.

[0033] Figure 2 a) is the nuclear magnetic resonance spectrum of monomer M1 in Example 1; b) is the nuclear magnetic resonance hydrogen spectrum of polymer P1 in Example 1.

[0034] Figure 3 is the synthetic route map and the nuclear magnetic resonance hydrogen spectrum of polymers P2 and P3 in Examples 2 and 3.

[0035] Figure 4 is the ultraviolet-visible light absorption spectrum of polymer P1 in Example 1 after complexing with three heavy metal ions of cadmium (Cd 2+ ), mercury (Hg 2+ ), and lead (Pb 2+ ).

[0036] Figure 5 is the fluorescence emission spectrum of polymer P1 in Example 1 after complexing with three heavy metal ions of Cd 2+ , Hg 2+ , and Pb 2+ .

[0037] Figure 6 a) is the cell safety evaluation result of polymer P1 in Example 1 at different concentrations; b) is the cell safety evaluation result of Cd 2+ ion at different concentrations.

[0038] Figure 7 is the maximum safe heavy metal Cd 2+ detoxification amount evaluation map of polymer P1 in Example 1.

[0039] Figure 8 is the survival curve of mice after intraperitoneal injection of Cd 2+ ion and intraperitoneal injection of polymer P1 to protect against Cd 2+ ion.

[0040] Figure 9 a) is the liver tissue HE pathological tissue section map of mice after intraperitoneal injection of polymer P1 to protect against Cd 2+ ion, b) is the kidney tissue HE pathological tissue section map of mice after intraperitoneal injection of polymer P1 to protect against Cd 2+ ion.

[0041] Figure 10 is the GPC outflow curve of polymer P1 in Example 1. DETAILED DESCRIPTION

[0042] Hereinafter, the present application will be described in detail. The description of the technical features described below is based on representative embodiments, specific examples of the present application, but the present application is not limited to these embodiments, specific examples.

[0043] <Definitions>

[0044] In the present specification, the term "alkyl group" includes straight, branched or cyclic alkyl groups, unless otherwise explicitly stated.

[0045] In the present specification, the term "hydrophilic group" means a group that imparts hydrophilicity to a compound or a polymer, and has a molecular weight of 300 or less, and the term "hydrophilic segment" means a molecular chain having a molecular weight of more than 300 composed of repeating units having hydrophilicity.

[0046] In the present specification, the numerical range represented by "numerical value A to numerical value B" means a range including the end point numerical values A and B.

[0047] In the present specification, the numerical range represented by "above" or "below" means a range including the present numerical value.

[0048] In the present specification, the meaning represented by "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0049] In the present specification, "optionally" or "optional" means that a certain substance, component, execution step, application condition, etc. is used or not used.

[0050] In the present specification, the unit names used are international standard unit names, and if not specifically stated, "%" used means a percentage by weight or mass.

[0051] In the present specification, the "preferred embodiments", "embodiments", and the like referred to mean that the particular element (e.g., feature, structure, property, and / or characteristic) described in connection with that embodiment is included in at least one embodiment described herein, and can or can not be present in other embodiments. In addition, it should be understood that the described elements can be combined in any suitable manner in various embodiments.

[0052] <Macromolecular Complexing Agent>

[0053] One of the objects of the present application is to provide a polymer containing structural unit I represented by formula I and structural unit II represented by formula II:

[0054]

[0055] In this context, R1 and R2 each independently represent an alkyl group having 1 to 10 carbon atoms or a phenyl group optionally substituted with an alkyl group, R3 and R4 each independently represent hydrogen or a methyl group, and Z represents a single bond, -O-, -S-, or -NR. 0 -, where R 0 It is a hydrogen or an alkyl group having 1 to 5 carbon atoms; B represents a single bond or an alkylene group having 1 to 10 carbon atoms; X represents a single bond, -O-, -S-, -CO-, or -NR. 0 -, -C(=O)NH-, -C(=O)O-, -OC(=O)- or alkylene groups with 1 to 10 carbon atoms, where G represents a hydrophilic group or hydrophilic segment.

[0056] In the polymers of the present invention, structural unit I, containing an aminophosphate structure in its side chain as shown in Formula I, endows the polymer with the ability to coordinate with heavy metals, and structural unit II, containing a hydrophilic group in its side chain as shown in Formula II, endows the polymer with water solubility. The polymers of the present invention, by simultaneously containing structural unit I and structural unit II, possess both good water solubility and heavy metal coordination ability.

[0057] The polymers of the present invention have a weight-average molecular weight of 1,000 to 500,000, preferably 5,000 to 200,000, more preferably 20,000 to 100,000. The number-average molecular weight is 500 to 300,000, preferably 2,000 to 80,000, more preferably 20,000 to 70,000. If the molecular weight is too low, the polymer's metabolism time is too fast, resulting in decreased safety; if the molecular weight is too high, the polymer's solubility in water decreases, viscosity increases, the rate of entry into cells slows down, and the efficacy decreases.

[0058] The polymer of the present invention has a molecular weight distribution of 1.1 to 3, for example 1.2 to 2.0.

[0059] In the polymer of the present invention, the molar ratio of structural unit I to structural unit II is (0.5-1.5):(0.5-10), preferably (0.8-1.2):(1-8). By keeping the molar ratio of these two structural units within the above range, both the water solubility and heavy metal complexing ability of the polymer can be taken into account.

[0060] The ratio of the content of structural unit I to structural unit II can be specifically selected depending on the molecular weight of the hydrophilic group or hydrophilic segment in structural unit II. In the case where structural unit II is a hydrophilic segment having a relatively large molecular weight such as a polyethylene glycol segment, a suitable ratio of the content of structural unit I to structural unit II is 1:(0.8 to 1.2), preferably 1:(0.9 to 1.1). In the case where structural unit II is a hydrophilic group having a relatively small molecular weight such as N,N-diethylacrylamide and 4-acryloylmorpholine, etc., a suitable ratio of the content of structural unit I to structural unit II is 1:(3 to 10), preferably 1:(4 to 8).

[0061] In one embodiment, the polymer is composed of structural unit I and structural unit II.

[0062] The following describes the structural units of the polymer of the present application, respectively.

[0063] Structural Unit I

[0064] In the polymer of the present application, structural unit I is represented by formula I, wherein the parameters appearing therein have one of the meanings given above and below. Unit I is derived from monomer 1 described below.

[0065] In one embodiment, Z represents a single bond, -O-, -S- or -NR 0 , preferably -O- or -NR 0 , wherein R 0 is hydrogen or alkyl having 1 to 5 carbon atoms, preferably hydrogen, methyl or ethyl.

[0066] In one embodiment, B represents a single bond or alkylene having 1 to 10 carbon atoms, preferably alkylene having 1 to 5 carbon atoms, such as methylene, ethylene, propylene, isopropylene, butylene, isobutylene, t-butylene, pentylene, isopentylene, neopentylene, cyclopentylene, hexylene, cyclohexylene, etc.

[0067] In one embodiment, R1and R2each independently represent alkyl having 1 to 5 carbon atoms, phenyl or alkyl-substituted phenyl, preferably R1and R2each independently represent methyl, ethyl, n-butyl, isobutyl, phenyl or benzyl.

[0068] In one embodiment, the number of repeating units of structural unit I in the molecular chain of the polymer of the present application is 1 to 300, preferably 5 to 150, more preferably 20 to 70.

[0069] In one embodiment, the ratio of the content of structural unit I in the polymer of the present application is 5 to 70 mol%, preferably 10 to 60 mol%, relative to the total number of moles of the structural units.

[0070] In a preferred embodiment, the structural unit I has the structure shown in the following formula (1):

[0071]

[0072] Structural Unit II

[0073] In the polymer of the present application, the structural unit II is shown in the following formula II, wherein the parameters appearing therein have one of the meanings given above and below. The unit II is derived from the following monomer 2.

[0074] In one embodiment, G is a hydroxyl group, a carboxyl group, a sulfonic acid group, a phosphoric acid group, a sulfuric acid group, an amide group, an amine group, a quaternary ammonium salt group, a morpholino group, or a polyethylene glycol segment, wherein a hydroxyl group, a carboxyl group, an amide group, an amine group, a morpholino group, or a polyethylene glycol segment is preferred, and a polyethylene glycol segment, an amine group, or a morpholino group is more preferred from the viewpoint of imparting better water solubility and safety to the polymer.

[0075] In one embodiment, the above-mentioned polyethylene glycol segment is an alkyl-terminated polyethylene glycol segment, wherein the terminal alkyl group is an alkyl group having 1 to 5 carbon atoms, preferably a methyl group, an ethyl group, or a propyl group, and more preferably a methyl group; and the number of repeating units of oxyethylene groups in the segment is 10 to 30, preferably 15 to 25.

[0076] In one embodiment, the above-mentioned amine group is a group represented by -NR'R", wherein R' and R" each independently represent hydrogen or an alkyl group having 1 to 10 carbon atoms, preferably hydrogen or an alkyl group having 1 to 5 carbon atoms, and more preferably hydrogen, a methyl group, an ethyl group, or a propyl group.

[0077] In one embodiment, the above-mentioned quaternary ammonium salt group is a trialkylammonium halide group, wherein the alkyl group has 1 to 5 carbon atoms, preferably a methyl group, an ethyl group, or a propyl group, and the halide counterion is a chloride ion or a bromide ion.

[0078] In one embodiment, X represents a single bond, -O-, -S-, -CO-, -NR 0 -, -C(=O)NH-, -C(=O)O-, -OC(=O)-, or an alkyl group having 1 to 10 carbon atoms, preferably a single bond, -CO-, -C(=O)NH-, or -C(=O)O-.

[0079] In the embodiment where G in formula II is a hydrophilic segment, the number of repeating units of the structural unit II in the molecular chain of the polymer of the present application is 1 to 300, preferably 5 to 150, and more preferably 20 to 70.

[0080] In the embodiment where G in formula II is a hydrophilic group, the number of repeating units of structural unit II in the molecular chain of the polymer of the present application is 1 to 2000, preferably 30 to 1000, more preferably 200 to 500.

[0081] In one embodiment, the polymer of the present application has a content ratio of structural unit II of 30 to 95 mole%, preferably 40 to 90 mole%, relative to the total number of moles of the structural units.

[0082] In one embodiment, structural unit II is a structural unit derived from (meth)acrylic acid, (meth)acrylamide, or polyethylene glycol monomethyl ether (meth)acrylate, N,N-diethyl (meth)acrylamide, (meth)acryloyl morpholine, or a vinyl alcohol structural unit.

[0083] In a preferred embodiment, structural unit II has one or more of the structures represented by formula (2), formula (3), or formula (4) below:

[0084]

[0085] Other Structural Units

[0086] The polymer of the present application also optionally contains other structural units, such as structural units derived from monomers selected from the group consisting of (meth)acrylates, vinyl acetate, styrenes.

[0087] In one embodiment, the polymer of the present application has a content ratio of other structural units of 20 mole% or less, preferably 10 mole% or less, more preferably 5 mole% or less, relative to the total number of moles of the structural units.

[0088] In one preferred embodiment, the polymer of the present application has the structure represented by formula (5) below:

[0089]

[0090] In formula (5), x and y each independently represent an integer of 1 to 300, preferably an integer of 5 to 150, more preferably an integer of 20 to 70, and n is 10 to 30, preferably 15 to 25.

[0091] In one preferred embodiment, the polymer of the present application has the structure represented by formula (6) below:

[0092]

[0093] In formula (6), x represents an integer of 1 to 300, preferably an integer of 5 to 150, more preferably an integer of 20 to 70; y represents an integer of 1 to 2000, preferably an integer of 30 to 1000, more preferably an integer of 200 to 500.

[0094] In one preferred embodiment, the polymer of the present application has a structure represented by the following formula (7):

[0095]

[0096] In formula (7), x represents an integer of 1 to 300, preferably an integer of 5 to 150, more preferably an integer of 20 to 70; y represents an integer of 1 to 2000, preferably an integer of 30 to 1000, more preferably an integer of 200 to 500.

[0097] <Preparation method>

[0098] One of the objects of the present application is to provide a preparation method of the polymer of the present application, comprising the steps of:

[0099] Step 1: reacting compound 1 or its hydrochloride salt, compound 2 and compound 3 in the presence of an acid-binding agent to obtain monomer 1;

[0100]

[0101] Step 2: polymerizing monomer 1 with monomer 2 and optionally other monomers in the presence of a free radical polymerization initiator,

[0102]

[0103] wherein each group has one of the meanings given in the context.

[0104] The individual steps of the preparation method of the present application are described in detail below.

[0105] Step 1

[0106] In step 1, monomer 1 is obtained by subjecting compound 1 or its hydrochloride salt, phosphite compound 2 and aromatic aldehyde compound 3 to Kabachnik-Fields reaction in the presence of an acid-binding agent.

[0107] The reaction of step 1 can be carried out in an organic solvent, and as the organic solvent, a mixed solvent of ethanol and acetonitrile (preferably in a volume ratio of 1:1), a dichloromethane solvent, an ethyl acetate solvent, a tetrahydrofuran solvent, etc. can be exemplified.

[0108] In one embodiment, the reaction temperature of the reaction of Step 1 is 35-80°C, preferably 50-70°C; the time is 2-12h, preferably 3-8h. In one embodiment, the control of the reaction temperature can be achieved by placing the reaction system in an oil bath.

[0109] In one embodiment, in Step 1, the molar ratio of Compound 1 or its hydrochloride, acid binding agent, Compound 3 to Compound 2 is (0.8-1.2):(0.8-1.2):(0.9-1.5):(1.0-2.0), preferably 1:1:1.1:1.2.

[0110] For the acid binding agent used, the present application is not particularly limited, which can be an acid binding agent known in the art for Kabachnik-Fields reaction, including but not limited to triethylamine, pyridine, 4-dimethylaminopyridine, etc.

[0111] In Step 1, the operation of separating and purifying the product of Kabachnik-Fields reaction is also optionally included.

[0112] In one embodiment, in Step 1, the operation of purifying the crude product by column chromatography after the end of the Kabachnik-Fields reaction, followed by the operation of rotary evaporation and freeze drying is also included. Specifically, in the column chromatography purification, the eluent can be a system of dichloromethane and methanol, and the volume ratio can be 20:1; the temperature of the rotary evaporation is 30-60°C, and the rotation speed is 0.5-10r / s; the temperature of the freeze drying is -20--50°C, the vacuum degree is 0.1-30Pa, the time is 12-72h, preferably 20-30h.

[0113] Step 2

[0114] In Step 2, the monomer 1 obtained in Step 1 is subjected to a free radical polymerization reaction with a water-soluble monomer 2 in the presence of a free radical polymerization initiator to obtain the polymer of the present application.

[0115] The free radical polymerization reaction of Step 2 is preferably carried out under anaerobic conditions, more preferably under anhydrous and anaerobic conditions. For the method of forming anaerobic conditions, it can be any method known in the art, for example, by the operation of multiple vacuum-inert gas introduction to the reaction vessel, bubbling inert gas into the reaction system or the operation of multiple liquid nitrogen freezing-vacuuming-thawing cycles. For the method of bubbling inert gas into the reaction system, the flow rate of the inert gas can be 10-100mL / min, and the bubbling time can be 5-60min.

[0116] In one embodiment, the radical polymerization reaction of Step 2 is preferably carried out in an anhydrous organic solvent, and N,N-dimethylformamide, dimethyl sulfoxide, and the like can be exemplified as the organic solvent.

[0117] In one embodiment, the reaction temperature of the radical polymerization reaction of Step 2 is 50 to 100°C, preferably 55 to 75°C, and the reaction time is 4 to 24 hours, preferably 8 to 16 hours, and the control of the reaction temperature can be achieved by placing the reaction system in an oil bath.

[0118] In one embodiment, the molar ratio of the radical initiator to monomer 1 in Step 2 is (0.01 to 0.1) : 1, and the molar ratio of monomer 1 to monomer 2 is (0.5 to 1.5) : (0.5 to 10), preferably (0.8 to 1.2) : (1 to 8).

[0119] In the case where G represents a hydrophilic segment, the molar ratio of monomer 1 to monomer 2 in Step 2 is 1 : (0.8 to 1.2), preferably 1 : (0.9 to 1.1).

[0120] In the case where G represents a hydrophilic segment, the molar ratio of monomer 1 to monomer 2 in Step 2 is 1 : (0.8 to 1.2), preferably 1 : (0.9 to 1.1).

[0121] The radical polymerization initiator used is not particularly limited in the present application, and can be any suitable radical polymerization initiator known in the art, including but not limited to azo-based initiators and peroxide initiators. For the azo-based initiators, azobisisobutyronitrile (abbreviated as AIBN), azobisisoheptyl nitrile (abbreviated as ABVN), and the like can be exemplified; and for the peroxide initiators, dibenzoyl peroxide (abbreviated as BPO), t-butyl hydroperoxide, and the like can be exemplified. These initiators can be used alone or in combination.

[0122] In one embodiment, Step 2 is to polymerize monomer 1, monomer 2, and other monomers in the presence of a radical polymerization initiator. The other monomer is not particularly limited in the present application, and can be any monomer known in the art that can be copolymerized with monomer 1 and monomer 2, including but not limited to (meth)acrylate monomers, vinyl acetate, styrene monomers, and the like.

[0123] Step 2 also optionally includes the operation of separating and purifying the reaction system after the radical polymerization reaction.

[0124] In one embodiment, after the radical polymerization reaction, the reaction solution is cooled, for example, the reaction vessel is cooled in ice water, and then the reaction solution is purified by dialysis in an organic solvent, followed by rotary evaporation and freeze-drying. Specifically, in the dialysis purification, the molecular weight cut-off of the dialysis membrane used is 1000-10000, and the dialysis time is 24-72 h; the organic solvent used in the dialysis purification is one or more selected from the group consisting of ethanol, methanol, acetonitrile, dichloromethane. Specifically, the temperature of the rotary evaporation is 30-60°C, and the rotation speed is 0.5-10 r / s. Specifically, the temperature of the freeze-drying is -20 to -50°C, the vacuum degree is 0.1-30 Pa, the time is 12-72 h, preferably 20-30 h.

[0125] <heavy metal antidote>

[0126] One of the objects of the present application is to provide a heavy metal antidote comprising the polymer of the present application and a pharmaceutically acceptable carrier.

[0127] The "pharmaceutically acceptable carrier" described in the present application can also be referred to as "excipient", "pharmaceutically acceptable adjuvant", "adjuvant" or "additional agent", etc., which refers to the general term of all additional materials used in the formulation of the prescription and the production of pharmaceuticals in addition to the active ingredient, which is generally a pharmaceutically acceptable inert ingredient that has been reasonably evaluated in terms of safety. The pharmaceutically acceptable carrier can enhance the handling properties of the pharmaceutical preparation, for example, by increasing the flowability and / or adhesiveness to make the preparation meet the process requirements. Further, the "pharmaceutically acceptable carrier" should have good compatibility with the active ingredient, i.e. the carrier itself or the impurities contained therein should not chemically react with the structural groups in the active ingredient or cause degradation of the active ingredient, resulting in a decrease in the content of the active ingredient.

[0128] Examples of the pharmaceutically acceptable carrier include, but are not limited to, solvents, binders, disintegrants, lubricating adjuvants (lubricants, glidants, anti-adhesion agents), stabilizers, fillers (or diluents), as well as flavoring agents, thickening agents, dispersing agents, coloring agents, bacteriostatic agents, antioxidants, pH adjusting agents, surfactants, spices and coating materials (plasticizers, light shielding agents, pigments) and the like.

[0129] In a preferred embodiment, the pharmaceutically acceptable carrier is a solvent, preferably water.

[0130] <use>

[0131] The present application also correspondingly relates to the use of the polymer of the present application for complexing or adsorbing heavy metals for non-therapeutic purposes.

[0132] The present application also correspondingly relates to the use of the polymer of the present application in the preparation of a medicament for the treatment of heavy metal poisoning.

[0133] Example

[0134] The following specific embodiments further illustrate the present invention. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the contents of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by this invention.

[0135] Example 1: Preparation of polymer P1

[0136] according to Figure 1 The synthetic route was used to prepare polymer P1.

[0137] N-(3-aminopropyl)methacrylamide hydrochloride (3.56 g, 20 mmol), triethylamine (2.02 g, 20 mmol), 8-hydroxyquinoline-2-carboxaldehyde (3.81 g, 22 mmol), and diethyl phosphite (3.31 g, 24 mmol) were dissolved in 10 mL of a 1:1 mixture of ethanol and acetonitrile. The mixture was reacted in an oil bath at 65 °C for 5 hours. After the reaction, the product was purified by column chromatography using dichloromethane and methanol as eluents (20:1 v / v). The monomer M1 was then collected by rotary evaporation and freeze-drying, resulting in a yellow viscous liquid with a yield of 74%.

[0138] Monomer M1 1 H-NMR spectrum as shown Figure 2 As shown in figure a), the target monomer M1 was successfully prepared.

[0139] Polyethylene glycol monomethyl ether methacrylate (9.5 g, 10 mmol), monomer M1 (4.35 g, 10 mmol), and azobisisobutyronitrile (50 mg, 0.2 mmol) were dissolved in 20 mL of N,N-dimethylformamide. After deoxygenation under nitrogen for 30 minutes, the mixture was reacted in an oil bath at 65 °C for 12 hours. After the reaction was completed, the reaction solution was cooled in ice water and purified by dialyzing in methanol solution using a dialysis bag with a molecular weight cutoff of 3500 for 48 hours. Polymer P1 was collected by rotary evaporation and freeze-drying as a light yellow solid powder with a yield of 82%. Polymer P1 was characterized by NMR.

[0140] Polymer P1 1 H-NMR spectrum as shown Figure 2 As shown in b), the target polymer P1 was successfully obtained.

[0141] The molecular weight of polymer P1 was determined by gel permeation chromatography (GPC) and the number average molecular weight was 48,000, the weight average molecular weight was 78,000, and the molecular weight distribution was 1.63. The elution curve of GPC is shown in FIG. 1. The GPC system was composed of a pump (Shimadzu LC-20AD), an automatic sampler, a guard column, an analytical column, and a differential refractive index detector (Shimadzu RID-10A). The mobile phase was DMF (1 mL / min), and the column temperature was 50°C. The system was calibrated using polystyrene standards with molecular weights of 200 to 10,000 g / mol. 6 1 Figure 10

[0142] Example 2: Preparation of polymer P2

[0143] N,N-diethylacrylamide (1.02 g, 8 mmol), monomer M1 (0.50 g, 2 mmol), and azobisisoheptanenitrile (25 mg, 0.1 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and after deoxygenation by nitrogen for 30 minutes, the reaction was placed in a 65°C oil bath for 12 hours. After the reaction was completed, the reaction solution was cooled in ice water, and the reaction solution was purified by dialysis in a methanol solution using a dialysis bag with a molecular weight cut-off of 3500 for 48 hours. The polymer P2 was collected as a light yellow solid powder by rotary evaporation and freeze-drying, with a yield of 78%. The polymer P2 was characterized by nuclear magnetic resonance.

[0144] The GPC spectrum of polymer P2 is shown in FIG. 2, and from the spectrum, it can be seen that the target polymer P2 was successfully obtained. Figure 3 Figure 3

[0145] The molecular weight of polymer P2 was determined by gel permeation chromatography (GPC) and the number average molecular weight was 32,000, the weight average molecular weight was 57,000, and the molecular weight distribution was 1.45.

[0146] Example 3: Preparation of polymer P3

[0147] 4-acryloylmorpholine (1.12 g, 8 mmol), monomer M1 (0.50 g, 2 mmol), and azobisisoheptanenitrile (25 mg, 0.1 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and after deoxygenation by nitrogen for 30 minutes, the reaction was placed in a 65°C oil bath for 12 hours. After the reaction was completed, the reaction solution was cooled in ice water, and the reaction solution was purified by dialysis in a methanol solution using a dialysis bag with a molecular weight cut-off of 3500 for 48 hours. The polymer P3 was collected as a light yellow solid powder by rotary evaporation and freeze-drying, with a yield of 81%. The polymer P3 was characterized by nuclear magnetic resonance.

[0148] The GPC spectrum of polymer P3 is shown in FIG. 3, and from the spectrum, it can be seen that the target polymer P3 was successfully obtained. 1 ​​​​​H-NMR spectrum is shown in Figure 3 As can be seen from Figure 3 , the target polymer P3 is successfully obtained.

[0149] The molecular weight of polymer P3 is determined by gel permeation chromatography (GPC), and the number average molecular weight is 37,000, the weight average molecular weight is 49,000, and the molecular weight distribution is 1.31.

[0150] Complexation Heavy Metal Ion Test

[0151] 1. UV-visible light absorption spectrum test

[0152] The polymer P1 obtained in Example 1 is dissolved in deionized water to prepare a polymer solution of 2 mg / mL. The polymer solution is mixed with 1 mg / mL aqueous solution of cadmium (Cd 2+ ), mercury (Hg 2+ ), and lead (Pb 2+ ) heavy metal ions in equal volume, and the complexation of the polymer with the three heavy metal ions is characterized by UV-visible light absorption spectrum, and the results are shown in Figure 4 .

[0153] Among them, the aqueous solution of cadmium (Cd 2+ ) ion is prepared by using cadmium nitrate tetrahydrate, and the aqueous solutions of mercury (Hg 2+ ) and lead (Pb 2+ ) ions are prepared by using standard solutions, as follows:

[0154] Lead standard solution 1000 ug / ml Item: GSB 04-1742-2004 Beijing Songyuan Kechuang Technology Co., Ltd.

[0155] Mercury standard solution 1000 ug / ml Item: 1821622-100 mL Shanghai Maikelin Biochemical Technology Co., Ltd.

[0156] As can be seen from Figure 4 , the absorption of polymer P1 combined with Hg 2+ is significantly enhanced at about 300 nm, the absorption of polymer P1 combined with Cd 2+ is enhanced to a certain extent at about 350 nm, and the absorption of polymer P1 combined with Pb 2+ is significantly enhanced at about 400 nm; the above results show that polymer P1 has good complexing ability with Cd 2+ , Hg 2+ , and Pb 2+ .

[0157] 2. Fluorescence emission spectrum test

[0158] The polymer P1 obtained in Example 1 was dissolved in deionized water to prepare a polymer solution of 2 mg / mL. The polymer solution was mixed with an aqueous solution of three heavy metal ions, Cd 2+ , Hg 2+ , Pb 2+ , each of 1 mg / mL, in equal volume, and the complexation of the polymer with the three heavy metal ions was characterized by fluorescence emission spectrum.

[0159] It can be seen from Figure 5 that the fluorescence of the polymer combined with Hg 2+ was significantly quenched, and the fluorescence was significantly red-shifted after being combined with Cd 2+ and Pb 2+ ; the above results further indicate that the polymer P1 has good complexing ability with Cd 2+ , Hg 2+ and Pb 2+ .

[0160] Safety Evaluation Test

[0161] The polymer P1 obtained in Example 1 was dissolved in 1640 medium at different concentrations (5 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL), and after co-culturing with mouse fibroblasts for 24 hours, the cells were repeatedly washed 3 times with PBS buffer, CCK-8 reagent (10%) was added, and the cells were further cultured for 2 hours. The absorbance at 450 nm wavelength was read by an enzyme-labeled instrument, and the safety of the polymer P1 was evaluated by the CCK-8 test method, and the results are shown in Figure 6 a). The safety of nitric acid cadmium was evaluated by the same method, and the results are shown in Figure 6 b).

[0162] It can be seen from Figure 6 a) that the polymer P1 has good safety, and the cell survival rate of 15 mg / mL polymer P1 is greater than 90%; it can be seen from Figure 6 b) that the toxicity of cadmium ions is very large, and the cell survival rate is greater than 90% only at a concentration of 2 μg / mL or less.

[0163] Cell Heavy Metal Detoxification Capacity Test

[0164] Based on the high toxicity of cadmium ions in the safety evaluation test, the safe concentration of polymer P1 (15 mg / mL, dissolved in 1640 medium) determined in the safety evaluation test was mixed with different concentrations (50 μg / mL, 100 μg / mL, 120 μg / mL, 140 μg / mL, 160 μg / mL, 180 μg / mL, 200 μg / mL, prepared by dissolving cadmium nitrate tetrahydrate in 1640 medium) of cadmium ions, and after 24 hours of co-culture with mouse fibroblasts, the cell safety was evaluated by CCK-8 test method to evaluate the maximum safe heavy metal detoxification amount of polymer P1 as a macromolecular detoxification agent, and the results are shown in Figure 7 .

[0165] From Figure 7 it can be seen that the maximum safe detoxification amount of polymer P1 for cadmium ions is about 160 μg / mL, while the safe concentration of cadmium ions is only 2 μg / mL, and polymer P1 can increase the safe dose of cadmium ions by 80 times, which indicates that polymer P1 has excellent effect as a macromolecular heavy metal detoxification agent.

[0166] Mouse Heavy Metal Detoxification Capacity Test

[0167] Cadmium nitrate tetrahydrate (50 μg of cadmium ions) was dissolved in 0.2 mL of normal saline and injected intraperitoneally into Balb / C mice to construct a mouse heavy metal poisoning model; after intraperitoneal injection of cadmium ions, 0.8 mL of 250 mg / mL polymer normal saline solution (250 mg of polymer dissolved in 1 mL of normal saline) was injected intraperitoneally into the poisoned mice 5 minutes later to construct a mouse heavy metal detoxification model; 0.2 mL of normal saline was injected as a control group to evaluate the heavy metal detoxification effect of polymer P1 as a macromolecular detoxification agent, and the results are shown in Figure 8 .

[0168] From Figure 8 it can be seen that intraperitoneal injection of 50 μg of cadmium ions will cause all Balb / C mice to die within 2 days, indicating that the mouse heavy metal model is successfully constructed. After intraperitoneal injection of cadmium ions for 5 minutes, further intraperitoneal injection of 0.8 mL of 250 mg / mL polymer normal saline solution, all mice survived for more than 7 days.

[0169] HE staining analysis of liver and kidney tissues of mice, the results are shown in Figure 9 a) liver tissue sections and Figure 9 b) kidney tissue sections, the liver and kidney tissue sections of the polymer protection group are consistent with those of healthy mice, and no pathological inflammation occurs, which indicates that polymer P1 has good detoxification effect as a macromolecular heavy metal detoxification agent.

[0170] Industrial applicability

[0171] The polymers of the present application can be widely used for complexing, adsorbing heavy metals and treating heavy metal poisoning.

Claims

1. A polymer, characterized in that, Contains structural unit I as shown in Equation I and structural unit II as shown in Equation II: In this context, R1 and R2 each independently represent an alkyl group having 1 to 10 carbon atoms or a phenyl group optionally substituted with an alkyl group, R3 and R4 each independently represent hydrogen or a methyl group, and Z represents a single bond, -O-, -S-, or -NR. 0 -, where R 0 It is a hydrogen or an alkyl group having 1 to 5 carbon atoms; B represents a single bond or an alkylene group having 1 to 10 carbon atoms; X represents a single bond, -O-, -S-, -CO-, or -NR. 0 -, -C(=O)NH-, -C(=O)O-, -OC(=O)- or alkylene groups with 1 to 10 carbon atoms, where G represents a hydrophilic group or hydrophilic segment.

2. The polymer according to claim 1, characterized in that, Its weight-average molecular weight is 1,000 to 500,000; the molar ratio of structural unit I to structural unit II is (0.5 to 1.5): (0.5 to 10).

3. The polymer according to claim 1 or 2, characterized in that, In Equation I, Z represents -O- or -NR. 0 - B represents an alkylene group with 1 to 10 carbon atoms; in Formula II, G represents a hydroxyl group, carboxyl group, sulfonic acid group, phosphate group, sulfate group, amide group, amino group, quaternary ammonium salt group, morpholino group or polyethylene glycol segment, and X represents a single bond, -CO-, -C(=O)NH- or -C(=O)O-.

4. The polymer according to claim 3, characterized in that, In Formula I, R1 and R2 each independently represent methyl, ethyl, n-butyl, isobutyl, phenyl, or benzyl, Z represents -NH-, and B represents an alkylene group with 1 to 5 carbon atoms; in Formula II, X represents -CO-, -CONH-, or -COO-; G is a polyethylene glycol segment, amino group, or morpholino group.

5. The method for preparing the polymer according to any one of claims 1 to 4, characterized in that, Includes the following steps: Step 1: React compound 1 or its hydrochloride salt, compound 2 and compound 3 in the presence of an acid-binding agent to obtain monomer 1; Step 2: Polymerize monomer 1 and monomer 2 in the presence of a free radical polymerization initiator; Each of the groups has the definition in any one of claims 1 to 4.

6. The preparation method according to claim 5, characterized in that, The acid-binding agent is triethylamine, pyridine, or 4-dimethylaminopyridine; the free radical polymerization initiator is one or more selected from azo initiators and peroxide initiators.

7. The preparation method according to claim 5 or 6, characterized in that, In step 1, the molar ratio of compound 1 or its hydrochloride salt, acid-binding agent, compound 3 and compound 2 is (0.8~1.2):(0.8~1.2):(0.9~1.3):(1.0~1.4); in step 2, the molar ratio of free radical polymerization initiator and monomer 1 is (0.01~0.1):1, and the molar ratio of monomer 1 and monomer 2 is (0.5~1.5):(0.5~10).

8. The preparation method according to claim 7, characterized in that, In step 1, the molar ratio of compound 1 or its hydrochloride salt, acid-binding agent, compound 3 and compound 2 is 1:1:1.1:1.

2.

9. The preparation method according to claim 5 or 6, characterized in that, The reaction temperature in step 1 is 35–80°C, and the reaction time is 2–12 h; the polymerization reaction in step 2 is carried out under anaerobic conditions, with a polymerization temperature of 50–100°C and a reaction time of 4–24 h.

10. A heavy metal detoxifying agent, characterized in that, It comprises the polymer according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier.

11. Use of the polymer according to any one of claims 1 to 4 in the preparation of a medicament for treating heavy metal poisoning.

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