Use of hemocompatible porous polymer bead sorbents to remove endotoxemia-inducing molecules.

Abstract

To provide a porous polymer based sorbent that can decrease endotoxin levels in biological fluids without potential leaching of harmful substances.SOLUTION: A biocompatible porous polymer based sorbent comprises at least one polymer, the polymer comprising a polyol or zwitterionic functional group designed to adsorb endotoxins and other inflammatory mediator molecules. The sorbent is applicable to the field of broadly reducing endotoxins in blood and blood products that can cause endotoxemia, and also to the field of broadly removing endotoxins by perfusion or hemoperfusion.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Related Applications

[0001] This application is a US Patent Application No. 62 / 341,676, filed May 26, 2016. No. 60 / 699,999, filed on Dec. 1, 2003, the disclosure of which is incorporated herein by reference in its entirety.

[0002] Technical Field

[0002] The disclosed invention is in the field of porous polymer-based sorbents. The disclosed invention is also in the field of broad reduction of endotoxins that can cause endotoxemia in blood and blood products. Furthermore, the disclosed invention is in the field of broad removal of endotoxins by perfusion or hemoperfusion. [Background technology]

[0003]

[0003] Gram-negative bacterial cell walls contain bound toxic substances known as endotoxins or lipopolysaccharides (LPS). Structurally, LPS consists of three distinct regions: the O antigen, the core, and lipid A. The O antigen is a repeating glycan polymer containing hydrophilic outermost domain molecules, the composition of which varies for each LPS lineage. The core links the O antigen to lipid A, a bioactive phosphorylated glucosamine disaccharide containing multiple hydrophobic fatty acid tails. These fatty acid tails are responsible for anchoring LPS within the bacterial cell wall. Both the core and lipid A are highly conserved across various LPS lineages, and lipid A is the major toxic component.

[0004]

[0004] There are two major routes by which endotoxin can enter the bloodstream and induce an inflammatory response in humans at intravenous doses as low as 1 ng / kg body weight / hour. The first is through local or systemic infection with foreign gram-negative bacteria, and the second is through translocation of endogenous gram-negative bacteria or their fragments across the intestinal membrane. Once in the circulation, LPS binds to lipopolysaccharide binding protein (LPB) to form LPS-LPB complexes, which can then induce an inflammatory response by triggering immune system and tissue cell responses. A prolonged upregulated inflammatory response can lead to sepsis or systemic inflammatory response syndrome (SIR). S), both of which can progress to potentially fatal septic shock and multiple organ dysfunction syndrome (MODS). There is a possibility that this will happen.

[0005] Endotoxins have also been associated with a myriad of syndromes and diseases, including complications from trauma, burns, and invasive surgical procedures, as well as organ-specific diseases such as liver disease, kidney dialysis complications, and autoimmune diseases.

[0006]

[0006] Currently, there are many commercially available endotoxin adsorbents. Several products based on polymyxin B (PMB) immobilized on agarose gel are available, including Detoxi-Gel Endotoxin Removing Gel (Thermo Fisher Scientific), AffiPrep Polymyxin Matrix (BioRad), Polymyxin B agarose (Sigma-Aldrich), and Endotoxin Affisorbent (bioWORLD). Toraymyxin (Toray Medical Co.) is a PMB-based extracorporeal device designed for selective endotoxin blood purification by direct hemoperfusion and has been approved as a therapeutic device by the Japanese Health Insurance System. Polymyxin B is characterized by a heptapeptide ring, a tripeptide group, and a fatty acid tail and is an antibiotic primarily used against resistant Gram-negative bacterial infections. The positively charged diaminobutyric acid group of PMB interacts with the negatively charged phosphate group of LPS, leading to an interaction between the N-terminal fatty acid chain of PMB and the lipid A fatty acid tail, forming a highly stable PMB-LPS complex (Harm, Stephan, Dieter Falkenhagen, and Jens Hartmann). “Endotoxin Adsorbents in Extracorporeal Blood Purification: Do They Fulfill Expectations?” Int J Artif Organs 37.3 (2014): 222-32.).

[0007]

[0007] Taking advantage of the negatively charged groups of LPS, anion exchange resins can also be used to remove LPS. Diethylaminoethyl-cellulose (DEAE-cellulose) resins are positively charged as a result of tertiary amine functional groups, and Bengsch et al. reported that DEAE-cellulose adsorbents bind endotoxin with high affinity and capacity in patients with sepsis. However, a transient but reversible prolongation of prothrombin time accompanied the reduction in endotoxin levels (Bengsch S, Boos KS, Nagel D, Seidel D, Inthorn D. Extracorporeal plasma treatment for the removal of endotoxin in patients with sepsis: clinical results of a pilot study. Shock. 2005; 23(6): 494-500.). The Alteco LPS adsorbent (Alteco Medical AB) consists of a polyethylene slab onto which the unique cationic peptide HAE 27 is immobilized, which selectively binds and adsorbs LPS. Furthermore, the EndoTrap (Profos AG) adsorbent consists of bacteriophage proteins immobilized on Sepharose beads, where the bacteriophage proteins have a high affinity for LPS molecules.

[0008] The adsorption capacity of a number of commercially available endotoxin adsorbents was assessed in a study by Harm et al. The adsorbents tested included Toraymyxin PMX-20R, Alteco LPS adsorbent, diethylaminoethyl-Sepharose (DEAE-Sepharose), Polymyxin B agarose, and EndoTrap red, and the mobile phases tested included buffer, protein solution, serum, heparinized plasma, and whole blood. Because only Alteco LPS Adsorber and Toraymyxin PMX-20R are hemocompatible, only these adsorbents were tested in whole blood. These two adsorbents are also the only two of the products designed for hemoperfusion applications. In batch adsorption studies using 10% adsorbent in a 100 ng FITC-LPS / mL solution, DEAE-Sepharose exhibited the best adsorption capacity compared with other materials tested, reducing LPS levels to 18 ± 8.5% of the control in 10 mM PBS buffer and 37 ± 4% of the control in a 4% (w / v) human serum albumin (HSA) solution. Toraymyxin was the only other adsorbent that reduced LPS activity by 70% and less than 95% in PBS and HSA solutions, respectively, resulting in a reduction of 21 ± 2% in PBS and 87 ± 6% in HSA. Batch studies were performed in serum and heparinized plasma using 10% adsorbent in serum or plasma spiked with 5 ng LPS / mL. DEAE-Sepharose was the most effective at removing LPS from serum, reducing levels to 28 ± 0.8% of control; however, DEAE-Sepharose's heparin-binding ability led to plasma clotting, making it unsuitable for testing in heparinized plasma. PMB-Agarose was the second most effective, reducing LPS levels to 36 ± 3.6% and 64 ± 6.8% of control in serum and heparinized plasma, respectively. Toraymyxin was the only other adsorbent able to reduce levels to 75% of control, reducing LAL activity to 41 ± 3.5% and 65 ± 4.5% of control in serum and heparinized plasma, respectively.In a batch study using 5% (w / v) adsorbent and an LPS concentration of 3 ng / mL in whole blood, Toraymyxin reduced activity to 60±14% of the control, whereas Alteco LPS Adsorber failed to reduce activity below 90%. (Harm, Stephan, Dieter Falkenhagen, and Jens Hartmann. “Endotoxin Adsorbents in Extracorporeal Blood Purification: Do They Fulfill Expectations?” Int J Artif Organs 37.3 (2014): 222-32.)

[0009] For hemoperfusion applications, a concern with the use of PMB immobilized on a polymeric support is the possibility that non-covalently bound PMB may leach from the support into the recirculating blood. Polymyxin B has been shown to induce neurotoxicity in some patients undergoing intravenous treatment (Weinstein, L, TL Doan, and MA Smith. "Neurotoxicity in patients treated with "Intravenous polymyxin B: Two case reports." Am J Health Syst Pharm 2009 Feb 15; 66(4): 345-7. Furthermore, PMB has been shown to induce nephrotoxicity in some patients undergoing intravenous treatment (Sobieszczyk, ME, et. al. "Combination therapy with polymyxin B for the treatment of multidrug-resistant Gram-negative respiratory tract infections." J Antimicrob Chemother. 2004 Aug; 54(2): 566-9). In the previously referenced study by Harm et al., noncovalently bound PMB from Toray fibers and PMB-agarose beads was separated by a series of washing steps and quantified using HPLC. The fibers or beads were incubated 10 times in saline, followed by five times in 0.1 N HCl solution. 42 ± 12 ng of PMB was obtained from Toray fibers after the fifth 0.1 N HCl washing step. From the PMB-Agarose beads, 27±6 ng PMB / mL was found after the fourth 0.1 N HCl washing step.

[0010] As mentioned above, endotoxins are components of the cell walls of Gram-negative bacteria. Gram-negative bacteria are commonly used for recombinant protein production, and many of the techniques used to extract desired recombinant proteins from bacterial cells also release lipopolysaccharides. Purification of recombinant proteins using ion exchange columns is not always completely successful because LPS tends to form complexes with proteins through specific or nonspecific interactions, resulting in the entire complex becoming immobilized on the exchange column. Ropp et al. developed a technique using alkanediols to separate LPS from protein-LPS complexes, leaving the protein immobilized on the ion exchange column (PCT Int. Appl. (2005), WO 2005003152 A1 20050113). Alkanediols were chosen because of their lower toxicity and inflammatory potential compared to other reagents that achieve similar separations.

[0011]

[0011] Furthermore, alkanediols exhibit broad-spectrum antimicrobial activity and are used as moisturizing antimicrobial agents in cosmetics. In the optimized structures of the dimer and trimer of 1,2-hexanediol and (S)-3-(hexyloxy)propane-1,2-diol in water, the proximity of two hydroxyl groups and the length of the aliphatic chain increase the amphiphilicity of the alkanediol, which may allow it to penetrate more easily into the membrane bilayer of microbial cells (Yoo IK, JII Kim, YK Kang. "Conformational preferences and antimicrobial activities of alkanediols." Computational and Theoretical Chemistry 2015 vol. 1064, 15-24.).

[0012] Lipoteichoic acid (LTA) is a major component of the cell wall of Gram-positive bacteria. It has many of the same pathogenic properties as LPS. LTA is anchored in the cell wall by glycolipids, which play a role similar to that of lipid A in LPS (Morath S, et al. "Structure / function relationships of lipoteichoic acids." J Endotoxin Res. 2005;11(6):348-56.). Upon dissociation from the cell wall, it binds nonspecifically to membrane phospholipids of target cells or specifically to toll-like receptors, activating the complement cascade or triggering the release of reactive species and cytokines, which may amplify cellular damage. LTA plays an important role in infections caused by Gram-positive bacteria, and in animal studies, it has been shown to trigger a cascade that leads to meningitis, encephalomyelitis, and arthritis as well as multiple organ failure and septic shock (Ginsburg I. "Role of lipoteichoic acid in infection and inflammation." Lancet Infect Dis. 2002 Mar;2(3):171-9.). [Prior art documents] [Patent documents]

[0013] [Patent Document 1] PCT Int. Appl. (2005), WO 2005003152 A1 20050113 [Non-patent literature]

[0014] [Non-Patent Document 1] Harm, Stephan, Dieter Falkenhagen, and Jens Hartmann. “Endotoxin Adsorbents in Extracorporeal Blood Purification: Do They Fulfill Expectations?” Int J Artif Organs 37.3 (2014): 222-32. [Non-patent document 2] Bengsch S, Boos KS, Nagel D, Seidel D, Inthorn D. Extracorporeal plasma treatment for the removal of endotoxin in patients with sepsis: clinical results of a pilot study. Shock. 2005; 23(6): 494-500. [Non-patent document 3] Weinstein, L, TL Doan, and MA Smith. “Neurotoxicity in patients treated with intravenous polymyxin B: Two case reports.” Am J Health Syst Pharm 2009 Feb 15; 66(4): 345-7. [Non-patent document 4] Sobieszczyk, ME, et. al. “Combination therapy with polymyxin B for the treatment of multidrug-resistant Gram-negative respiratory tract infections.” J Antimicrob Chemother. 2004 Aug; 54(2): 566-9. [Non-Patent Document 5] Yoo IK, JII Kim, YK Kang. “Conformational preferences and antimicrobial activities of alkanediols.” Computational and Theoretical Chemistry 2015 vol 1064, 15-24. [Non-patent document 6] Morath S, et. al. “Structure / function relationships of lipoteichoic acids. J Endotoxin Res. 2005; 11(6): 348-56. [Non-Patent Document 7] Ginsburg I. “Role of lipoteichoic acid in infection and inflammation.” Lancet Infect Dis. 2002 Mar; 2(3): 171-9. Summary of the Invention

[0015] The novel sorbent materials described herein offer advantages over existing technologies in that they reduce endotoxin levels in biological fluids without the potential for leaching of harmful substances, resulting in a safe and effective method. The net neutral charge of the functional groups covalently attached to the polymer matrix makes these sorbents different from other existing technologies. LPS can be retained by the novel sorbent materials through tortuous pathways, sorption, and pore entrapment. Several routes are available for synthesizing the resins, which contain either polyol or zwitterionic groups covalently attached to the poly(styrene-co-divinylbenzene) backbone. For hemoperfusion applications, hemocompatibility of the polymer is a requirement. Using the unactivated partial thromboplastin time (uPTT) assay as a measure of thrombogenicity, the polymers described herein exhibit minimal activation, which is indicative of plasma-like interactions. Furthermore, these sorbents can simultaneously remove cytokines and inflammatory protein moieties while removing endotoxins, and potentially exhibit antimicrobial activity. Removing either endotoxins or cytokines from endotoxemic patients may be an insufficient treatment, as residual endotoxins may trigger further cytokine production, and residual cytokines may still lead to sepsis. By removing both the source of infection and the subsequent excessive inflammatory response, this novel sorbent offers advantages over existing technologies specifically designed for endotoxin removal.

[0016] In one aspect, the present invention relates to a biocompatible polymer system comprising at least one polymer, the polymer being either a polyol or a zwitterionic functional group. The polymer systems include any of the above; the polymer systems are capable of adsorbing endotoxins. Preferred polymers are also capable of adsorbing a wide range of toxins and inflammatory mediators having molecular weights from less than about 0.5 kDa to about 1,000 kDa (or in some embodiments, from about 1 kDa to about 1,000 kDa). Some preferred polymers are hemocompatible. Some preferred polymer systems have a spherical bead geometry.

[0017]

[0015] Certain preferred polymers are also capable of adsorbing one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, including but not limited to lipopolysaccharide (LPS). Other preferred polymers are also capable of adsorbing one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, including but not limited to lipoteichoic acid (LTA). In some embodiments, toxins and inflammatory mediators include one or more of the following: cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies.

[0018] The polymer can be made by any means known in the art for making suitable porous polymers. In one preferred embodiment, the polymer is made using suspension polymerization. In other embodiments, the polymer is made by emulsion polymerization, bulk polymerization, or precipitation polymerization.

[0019]

[0017] The polymer is in the form of a solid support. In some preferred embodiments, the solid support is a bead. In other embodiments, the solid support is a fiber, a monolithic column, or a film.

[0020] Some polymer systems have a polymer pore structure with a total volume of greater than 0.1 cc / g and less than 5.0 cc / g (dry polymer) for pore sizes from 10 Å to 40,000 Å, while other polymer systems are non-porous. Other embodiments have a polymer pore structure with a total volume of greater than 0.1 cc / g and less than 3.0 cc / g (dry polymer) for pore sizes from 10 Å to 40,000 Å, while other polymer systems are non-porous.

[0021] In some embodiments, the polymer is in the form of hypercrosslinked or macroreticular porous polymer beads containing polyol groups. In some other embodiments, the polymer is in the form of hypercrosslinked or macroreticular porous polymer beads containing zwitterionic groups. In a preferred embodiment, the polymer is in the form of hypercrosslinked or macroreticular porous polymer beads containing diol groups.

[0022] In some embodiments, the polymer is in the form of non-porous polymer beads containing polyol groups. In some other embodiments, the polymer is in the form of non-porous polymer beads containing zwitterionic groups. In a preferred embodiment, the polymer is in the form of non-porous polymer beads containing diol groups.

[0023] In some embodiments, the polymeric beads contain polyol groups. Polymeric beads containing polyol groups can be prepared by ring-opening reactions of preformed polymers containing epoxide groups. In a preferred embodiment, the polyol groups are diol groups.

[0024] In another embodiment, polymeric beads containing polyol groups can be prepared by the ester hydrolysis reaction of preformed polymers containing residual acetate groups. In a preferred embodiment, the polyol groups are diol groups.

[0025] In certain other embodiments, the polymeric beads contain zwitterionic functional groups. Polymeric beads containing zwitterionic functional groups can be prepared by free radical reactions in the presence of zwitterionic monomers that contain double bonds that are readily available for polymerization.

[0026]

[0024] Some polymer systems are constructed from polymerizable vinyl monomers containing epoxide groups and are combined with a crosslinker, a hemocompatible monomer, a monomer, and a suitable porogen in the presence of the monomer. The epoxides are then copolymerized in the presence of a base to yield porous polymers containing epoxide functional groups. These epoxides are then converted to polyols by a ring-opening reaction in the presence of a base. In a preferred system, the epoxides are converted to diols.

[0027]

[0025] Still other polymer systems are constructed from polymerizable vinyl monomers containing acetate groups, which are copolymerized in the presence of a crosslinker, a hemocompatible monomer, a monomer, and a suitable porogen to obtain porous polymers containing acetate groups. These acetate groups are converted to polyols by ester hydrolysis in the presence of a base. In a preferred embodiment, the polyol groups are diol groups.

[0028]

[0026] Some polymer systems are constructed from polymerizable vinyl monomers containing epoxide groups, which are copolymerized in the presence of a crosslinker, a hemocompatible monomer, and a monomer to obtain nonporous polymerized polymers containing epoxide functional groups. These epoxides are then converted into polyols by ring-opening reaction in the presence of a base. In a preferred system, the epoxides are converted into diols.

[0029]

[0027] Another polymer system is constructed from polymerizable vinyl monomers containing acetate groups, which are copolymerized in the presence of a crosslinking agent, a hemocompatible monomer, and a monomer to obtain a non-porous polymer containing acetate groups. These acetate groups are converted to polyols by ester hydrolysis in the presence of a base. In a preferred embodiment, the polyol groups are diol groups.

[0030]

[0028] Some polymers are formed and then modified to become biocompatible. Some modifications include forming a biocompatible surface coating or layer. Yet another aspect relates to a device for removing endotoxins from physiological fluids, comprising the biocompatible polymer system described herein. Another aspect relates to a device for removing a wide range of protein-based toxins, from less than 0.5 kDa to 1,000 kDa, from physiological fluids, comprising the biocompatible polymer system described herein.

[0031] Another aspect relates to a device for removing one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components from a physiological fluid, comprising a biocompatible polymer system as described herein. Yet another aspect relates to a device for removing one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components from a physiological fluid, comprising a biocompatible polymer system as described herein.

[0032] Yet another aspect relates to a device for removing endotoxins from non-physiological fluids, comprising the biocompatible polymer system described herein. Another aspect relates to a device for removing a wide range of protein-based toxins, from less than 0.5 kDa to 1,000 kDa, from non-physiological fluids, comprising the biocompatible polymer system described herein.

[0033] Another aspect relates to a device for removing one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components from non-physiological fluids, comprising a biocompatible polymer system as described herein. Yet another aspect relates to a device for removing one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components from non-physiological fluids, comprising a biocompatible polymer system as described herein.

[0034] Another aspect includes a method of perfusion comprising passing physiological fluid through a device containing the biocompatible polymer system described herein, one or multiple times, via a suitable extracorporeal circuit.

[0035] Yet another aspect relates to enteral or rectal administration applications of the polymers described herein. In one aspect, the present invention relates to a non-biocompatible polymer system comprising at least one polymer, the polymer comprising either a polyol or a zwitterionic functional group; the polymer system is capable of adsorbing endotoxins from physiological fluids, laboratory or manufacturing fluids, or aqueous systems in one or more of health care facilities, home health care applications, medical facilities, biotechnology facilities, biological manufacturing processes, cell culture manufacturing processes, and laboratories. Preferred polymers are also capable of adsorbing one or more of a wide range of toxins, bacteria, bacterial fragments, and bacterial components. Aspects of the present invention also include the following. Aspect 1 A biocompatible polymer system comprising at least one polymer, wherein the polymer comprises either a polyol or a zwitterionic functional group; and wherein the polymer system is capable of adsorbing endotoxins. Aspect 2 2. The biocompatible polymer system according to embodiment 1, wherein the polymer system is also capable of adsorbing a wide range of toxins and inflammatory mediators. Aspect 3 3. The biocompatible polymer system of embodiment 2, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 1,000 kDa. Aspect 4 3. The biocompatible polymer system of embodiment 2, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 60 kDa. Aspect 5 3. The biocompatible polymer system of embodiment 2, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies. Aspect 6 2. The biocompatible polymer system according to embodiment 1, wherein the polymer system is also capable of adsorbing one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS). Aspect 7 2. The biocompatible polymer system according to embodiment 1, wherein the polymer system is also capable of adsorbing one or more of Gram-positive bacteria, Gram-positive bacterial fragments, and Gram-positive bacterial components, such as lipoteichoic acid (LTA). Aspect 8 2. The biocompatible polymer system of embodiment 1, wherein the polymer is produced using suspension polymerization, emulsion polymerization, bulk polymerization, or precipitation polymerization. Aspect 9 The polymers are created by modification of cellulosic polymers, and the modification is carried out by free radicals or S N 2. A biocompatible polymer system according to embodiment 1, comprising polyol or zwitterionic substrates attached by two types of chemistry, and optionally the attachment of lipophilic substrates comprising aryl or alkyl groups. Aspect 10 2. The biocompatible polymer system of embodiment 1, wherein the polymer system has the form of a solid support, which may include, but is not limited to, a bead, a fiber, a monolith column, a film, a membrane, or a semipermeable membrane. Aspect 11 11. The biocompatible polymer system according to embodiment 10, wherein the solid support has a biocompatible hydrogel coating. Aspect 12 2. The biocompatible polymer system of embodiment 1, wherein the polymer comprises a large number of pores, and the pore structure of the polymer has a total volume of greater than 0.1 cc / g and less than 5.0 cc / g (dry polymer) for pore sizes ranging from 10 Å to 40,000 Å. Aspect 13 2. The biocompatible polymer system of embodiment 1, wherein the polymer is non-porous. Aspect 14 2. The biocompatible polymer system according to embodiment 1, wherein the polymer is a highly crosslinked polymer. Aspect 15 2. The biocompatible polymer system according to embodiment 1, wherein the polymer is hemocompatible. Aspect 16 2. The biocompatible polymer system of embodiment 1, wherein the agent used to confer biocompatibility is either (i) heparin or (ii) a heparin-mimetic polymer. Aspect 17 2. The biocompatible polymer system of embodiment 1, wherein the polymer is formed and then modified to become biocompatible. Aspect 18 The modification that confers biocompatibility according to embodiment 17, wherein the agent used to confer biocompatibility is either (i) heparin or (ii) a heparin-mimetic polymer. Aspect 19 A device for removing endotoxins from physiological fluids, comprising the biocompatible polymer system according to any one of embodiments 1 to 18. Aspect 20 20. The device of embodiment 19, wherein the device also removes a broad range of toxins and inflammatory mediators. Aspect 21 21. The device of embodiment 20, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 1,000 kDa. Aspect 22 21. The device of embodiment 20, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 60 kDa. Aspect 23 21. The device of embodiment 20, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies. Aspect 24 20. The device of embodiment 19, wherein the device also removes one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS). Aspect 25 20. The device of embodiment 19, wherein the device also removes one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, such as lipoteichoic acid (LTA). Aspect 26 A device for removing endotoxins from non-physiological fluids, comprising the biocompatible polymer system according to any one of embodiments 1 to 18. Aspect 27 27. The device of embodiment 26, wherein the device also removes a broad range of toxins and inflammatory mediators. Aspect 28 28. The device of embodiment 27, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 1,000 kDa. Aspect 29 28. The device of embodiment 27, wherein the toxin and inflammatory mediator have a molecular weight of less than about 0.5 kDa to about 60 kDa. Aspect 30 28. The device of embodiment 27, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies. Aspect 31 27. The device of embodiment 26, wherein the device also removes one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS). Aspect 32 27. The device of embodiment 26, wherein the device also removes one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, such as lipoteichoic acid (LTA). Aspect 33 19. The biocompatible polymer system according to any of embodiments 1-18, in a device suitable for holding the polymer and suitable for incorporation into an extracorporeal circuit. Aspect 34 A method of perfusion comprising passing a physiological fluid through a device containing a biocompatible polymer system according to any one of aspects 1 to 18, one or more times, by means of a suitable extracorporeal circuit. Aspect 35 19. The biocompatible polymer system of any of embodiments 1 to 18, contained in a container suitable for holding the polymer for transfusion of a blood product comprising whole blood, packed red blood cells, platelets, albumin, plasma, or any combination thereof. Aspect 36 19. The biocompatible polymer system according to any of embodiments 1 to 18, for removing endotoxins from blood products including whole blood, plasma or serum, or from other physiological fluids. Aspect 37 19. The biocompatible polymer system according to any of embodiments 1 to 18, wherein the polymer is administered enterally or rectally. Aspect 38 A polymer system comprising at least one polymer, wherein the polymer comprises either a polyol or a zwitterionic functional group; and wherein the polymer system is capable of adsorbing endotoxin. Aspect 39 39. The polymer system of embodiment 38, wherein the polymer system is also capable of adsorbing a wide range of toxins, gram-negative bacteria, gram-negative bacterial fragments, gram-negative bacterial components such as lipopolysaccharide (LPS), gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components such as lipoteichoic acid (LTA). Aspect 40 40. The polymer system of embodiment 39, wherein the toxin has a molecular weight of from less than about 0.5 kDa to about 1,000 kDa. [Brief explanation of the drawings]

[0036]

[0035] The accompanying drawings, which are used to provide a further understanding of the present disclosure, are incorporated in and constitute a part of this specification, and together with the detailed description showing aspects of the present disclosure, are intended to explain the principles of the present disclosure. No structural details of the present disclosure are intended to be more detailed than is deemed necessary for a basic understanding of the present disclosure and various ways in which it can be implemented. The drawings show: [Figure 1] FIG. 1 shows the log differential pore volume plot for the modified polymer. [Figure 2] FIG. 2 shows the log differential pore volume plot for the modified polymer. [Figure 3] FIG. 3 shows the log differential pore volume plot for the modified polymer. [Figure 4] FIG. 4 shows the log differential pore volume plot for the modified polymer. [Figure 5]

[0037] Figure 5 shows endotoxin removal data from the kinetic model in human plasma for modified polymers CY15129, CY15154, and CY16000, expressed as a percentage determined by the amount of endotoxin remaining after 120 minutes compared to the pre-circulation concentration. [Figure 6]

[0038] Figure 6 shows endotoxin clearance data from the kinetic model in human plasma for polymer CY15154 and its unmodified precursor CY15077, expressed as a percentage determined by the amount of endotoxin remaining after 120 minutes compared to the pre-circulating concentration. [Figure 7]

[0039] FIG. 7 shows cytokine clearance data from the dynamic model in whole blood, expressed as a percentage determined by the amount of cytokine remaining at a particular time point compared to the pre-circulating concentration. DETAILED DESCRIPTION OF THE INVENTION

[0037]

[0040] Detailed embodiments of the present invention are disclosed herein as necessary; it should be understood that the disclosed embodiments are merely exemplary of the present invention, which can be embodied in various embodiments. Therefore, the specific structural and functional details disclosed herein should not be construed as limitations, but merely as a basis for teaching those skilled in the art how to use the present invention. The present invention may be better understood by the following specific examples, which are provided for guidance only and are not intended to be limiting in any way.

[0038]

[0041] The present invention may be more readily understood by reference to the following detailed description in conjunction with the accompanying drawings and examples, which form a part of this disclosure. It is to be understood that the present invention is not limited to the particular materials, devices, methods, applications, conditions, or parameters described and / or suggested herein, and that the terminology used herein is for the purpose of describing particular embodiments, by way of example, only, and is not intended to limit the invention as defined by the claims. As used herein, the term "plurality" means more than one. When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value constitutes another embodiment. All ranges are inclusive and combinable.

[0039]

[0042] For example, it should be recognized that certain features of the invention that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination. Furthermore, values ​​stated in ranges include each and every value and combination of values ​​within that range.

[0040]

[0043] The following definitions are provided to aid in the understanding of the present invention.

[0044] The term "biocompatible" means that the sorbent is biocompatible with physiological fluids, biological tissues, or "Sorbent" is defined to mean that the sorbent material is capable of contacting physiological fluids, living tissues, or organisms without producing unacceptable clinical changes during the time that the sorbent is in contact with the physiological fluids, living tissues, or organisms.

[0041]

[0045] The term "hemocompatible" is defined as a condition in which a biocompatible material undergoes a clinically acceptable physiological change when contacted with whole blood or plasma.

[0046] As used herein, the term "physiological fluid" refers to fluids derived from the body. The fluids that are involved in the production of blood include, but are not limited to, nasopharyngeal fluid, oral fluid, esophageal fluid, gastric fluid, pancreatic fluid, hepatic fluid, pleural fluid, pericardial fluid, peritoneal fluid, intestinal fluid, prostatic fluid, semen, vaginal secretions, as well as tears, saliva, lung or bronchial secretions, mucus, bile, blood, lymph, plasma, serum, synovial fluid, cerebrospinal fluid, urine, and interstitial, intracellular, and extracellular fluids, such as fluids exuding from burns or wounds.

[0042]

[0047] As used herein, the term "laboratory or manufacturing fluid" is defined as a liquid used in life science applications and can include, but is not limited to, tissue culture and cell culture media and additives, chemical and biological assay media, sample preparation buffers, biological manufacturing media, growth media, and bioreactor media.

[0043]

[0048] As used herein, the term "sorbent" includes adsorbents and absorbents. It can be enjoyed.

[0049] For purposes of this invention, the term "sorb" is defined as "to take up and bind by absorption and adsorption."

[0044]

[0050] For purposes of this invention, the term "perfusion" is defined as the single passage of physiological fluid through a suitable extracorporeal circuit through a device containing a porous polymeric adsorbent material to remove toxic molecules from the fluid.

[0045]

[0051] The term "hemoperfusion" refers to a specific example of perfusion in which the physiological fluid is blood. is.

[0052] The term "dispersant" or "dispersing agent" is defined as a substance that imparts a stabilizing effect to an array of finely divided immiscible droplets suspended in a fluidizing agent. do.

[0046]

[0053] The term "heparin-mimetic polymer" refers to any polymer that has the same anticoagulant and / or antithrombogenic properties as heparin.

[0054] The term "macroreticular synthesis" is defined as the polymerization of monomers into polymers in the presence of an inert precipitant that extrudes the growing polymer molecules from the monomer liquid at specific molecular sizes governed by phase equilibrium, resulting in spherical or nearly spherical, symmetrically packed solid nano-sized microgel particles that form beads with open-cell physical pores. [US Patent 4,297,220, Meitzner and Oline, October 27, 1981; R.L. Albright, Reactive Polymers, 4, 155-174(1986)].

[0047]

[0055] The term "hypercrosslinked" describes polymers in which a single repeat unit has more than two connectivity. Hypercrosslinked polymers are produced by crosslinking swollen or dissolved polymer chains with multiple robust crosslink spacers rather than by copolymerization of monomers. Crosslinkers can include bis(chloromethyl) derivatives of aromatic hydrocarbons, methylal, monochlorodimethyl ether, and other bifunctional compounds, which react with polymers in the presence of Friedel-Crafts catalysts [Tsyurupa, MP, ZK Blinnikova, NA Proskurina, AV Pastukhov, LA Pavlova, and VA Davankov. "Hypercrosslinked Polystyrene: The First Nanoporous Polymeric Material." Nanotechnologies in Russia 4 (2009): 665-75.]

[0048]

[0056] Some preferred polymers comprise residues from one or more monomers selected from the following, or comprise monomers or mixtures thereof: acrylonitrile, allyl glycidyl ether, butyl acrylate, butyl methacrylate, cetyl acrylate, cetyl methacrylate, 3,4-dihydroxy-1-butene, dipentaerythritol diacrylate, dipentaerythritol dimethacrylate, dipentaerythritol tetraacrylate, dipentaerythritol tetramethacrylate, dipentaerythritol triacrylate, dipentaerythritol trimethacrylate, divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone, 3,4-epoxy-1-butene, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, Ethyl acrylate, ethyl methacrylate, ethylstyrene, ethylvinylbenzene, glycidyl methacrylate, methyl acrylate, methyl methacrylate, octyl acrylate, octyl methacrylate, pentaerythritol diacrylate, pentaerythritol dimethacrylate, pentaerythritol tetraacrylate, pentaerythritol tetramethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, styrene, trimethylolpropane diacrylate, trimethylolpropane dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, trivinylbenzene, trivinylcyclohexane, vinyl acetate, vinylbenzyl alcohol, 4-vinyl-1-cyclohexene 1,2-epoxide, vinylformamide, vinylnaphthalene, 2-vinyloxirane, and vinyltoluene.

[0049]

[0057] Some embodiments of the present invention use organic solvents and / or polymeric porogens as porogens or pore-forming agents to induce phase separation during polymerization to produce porous polymers. Some preferred porogens are mixtures selected from the following, or any combination thereof: benzyl alcohol, cyclohexane, cyclohexanol, cyclohexanone, decane, dibutyl phthalate, di-2-ethylhexyl phthalate, di-2-ethylhexyl phosphate, ethyl acetate, 2-ethyl-1-hexanoic acid, 2-ethyl-1-hexanol, n-heptane, n-hexane, isoamyl acetate, isoamyl alcohol, n-octane, pentanol, poly(propylene glycol), polystyrene, poly(styrene-co-methyl methacrylate), tetralin, toluene, tri-n-butyl phosphate, 1,2,3-trichloropropane, 2,2,4-trimethylpentane, and xylene.

[0050]

[0058] In still other embodiments, the dispersing agent is selected from a mixture consisting of: hydroxyethyl cellulose, hydroxypropyl cellulose, poly(diethylaminoethyl acrylate), poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl acrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(vinyl alcohol), salts of poly(acrylic acid), salts of poly(methacrylic acid), and mixtures thereof.

[0051]

[0059] Preferred sorbents are biocompatible. In other further embodiments, the polymer is biocompatible. In yet other embodiments, the polymer is hemocompatible. In yet still further embodiments, the biocompatible polymer is hemocompatible. In yet still further embodiments, the polymer's geometric shape is a spherical bead.

[0052]

[0060] In other embodiments, the biocompatible polymer comprises poly(N-vinylpyrrolidone).

[0061] In other embodiments, the biocompatible polymer comprises a 1,2-diol. In other embodiments, the biocompatible polymer comprises a 1,3-diol.

[0053]

[0062] In other further embodiments, the biocompatible polymer comprises a heparin-mimetic polymer.

[0063] The coating / dispersion on the poly(styrene-co-divinylbenzene) resin will impart improved biocompatibility to the material.

[0054]

[0064] In still other embodiments, the following group of crosslinkers can be used to form the hemocompatible hydrogel coating: dipentaerythritol diacrylate, dipentaerythritol dimethacrylate, dipentaerythritol tetraacrylate, dipentaerythritol tetramethacrylate, dipentaerythritol triacrylate, dipentaerythritol trimethacrylate, divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone, pentaerythritol diacrylate, pentaerythritol dimethacrylate, pentaerythritol tetraacrylate, pentaerythritol tetramethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, trimethylolpropane diacrylate, trimethylolpropane dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, trivinylbenzene, trivinylcyclohexane, and mixtures thereof.

[0055]

[0065] In some embodiments, the polymer comprises at least one crosslinker and at least one dispersing agent. The dispersing agent may be biocompatible. The dispersing agent may be selected from chemicals, compounds, or substances such as hydroxyethyl cellulose, hydroxypropyl cellulose, poly(diethylaminoethyl acrylate), poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl acrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(vinyl alcohol), salts of poly(acrylic acid), salts of poly(methacrylic acid), and mixtures thereof; the crosslinker may be selected from the group consisting of dipentaerythritol diacrylate, dipentaerythritol dimethacrylate, dipentaerythritol tetraacrylate, and dipentaerythritol tetramethacrylate. dipentaerythritol triacrylate, dipentaerythritol trimethacrylate, divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone, pentaerythritol diacrylate, pentaerythritol dimethacrylate, pentaerythritol tetraacrylate, pentaerythritol tetramethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, trimethylolpropane diacrylate, trimethylolpropane dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, trivinylbenzene, trivinylcyclohexane, and mixtures thereof. Preferably, the polymer is developed simultaneously with the formation of the coating, whereupon the dispersing agent becomes chemically bonded to or entangled with the surface of the polymer.

[0056]

[0066] In still other embodiments, the biocompatible polymer coating is selected from the group consisting of poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(N-vinylpyrrolidone), poly(vinyl alcohol), salts of poly(acrylic acid), salts of poly(methacrylic acid), and mixtures thereof.

[0057]

[0067] In still other embodiments, the biocompatible oligomeric coating is selected from the group consisting of poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(N-vinylpyrrolidone), poly(vinyl alcohol), salts of poly(acrylic acid), salts of poly(methacrylic acid), and mixtures thereof.

[0058]

[0068] Certain biocompatible sorbent compositions of the present invention are composed of a multitude of pores. The biocompatible sorbent is designed to adsorb a wide range of toxins, from less than 0.5 kDa to 1,000 kDa. Without wishing to be bound by theory, it is believed that the sorbent works by sequestering molecules of a predetermined molecular weight within the pores. The size of molecules that a polymer can sorb will increase as the pore size of the polymer increases. Conversely, as the pore size exceeds the optimal pore size for adsorption of a particular molecule, adsorption of that protein can be or will decrease.

[0059]

[0069] In some methods, the solid form is porous. Some solid forms are characterized by a polymeric pore structure with a total volume of greater than 0.1 cc / g and less than 5.0 cc / g (dry polymer) of pore sizes ranging from 10 Å to 40,000 Å.

[0060]

[0070] In some other methods, the solid form is non-porous.

[0071] In some embodiments, the polymers can be made into beads with diameters ranging from 0.1 micrometers to 2 centimeters. Some polymers are powders, beads, or other regularly or irregularly shaped particles.

[0061]

[0072] In some embodiments, the multiple solid forms comprise particles having diameters ranging from 0.1 micrometers to 2 centimeters.

[0073] In some methods, the undesired molecules include endotoxins, gram-negative bacteria, gram-negative bacterial fragments, gram-negative bacterial components, gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, as well as inflammatory mediators and stimulators including cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, and peptides including bradykinin. Desiccant, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies.

[0062]

[0074] In certain embodiments, the sorbent material includes a crosslinked polymeric material derived from the reaction of a crosslinker with one or more of the following polymerizable monomers, followed by epoxidation and ring-opening to form a polyol: acrylonitrile, allyl glycidyl ether, butyl acrylate, butyl methacrylate, cetyl acrylate, cetyl methacrylate, 3,4-dihydroxy-1-butene, dipentaerythritol diacrylate, dipentaerythritol dimethacrylate, dipentaerythritol tetraacrylate, dipentaerythritol tetramethacrylate, dipentaerythritol triacrylate, dipentaerythritol trimethacrylate, divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone, 3,4-epoxy-1-butene, 1,2-epoxy-9-decene, 1,2- Epoxy-5-hexene, ethyl acrylate, ethyl methacrylate, ethylstyrene, ethylvinylbenzene, glycidyl methacrylate, methyl acrylate, methyl methacrylate, octyl acrylate, octyl methacrylate, pentaerythritol diacrylate, pentaerythritol dimethacrylate, pentaerythritol tetraacrylate, pentaerythritol tetramethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, styrene, trimethylolpropane diacrylate, trimethylolpropane dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, trivinylbenzene, trivinylcyclohexane, vinyl acetate, vinylbenzyl alcohol, 4-vinyl-1-cyclohexene 1,2-epoxide, vinylformamide, vinylnaphthalene, 2-vinyloxirane, and vinyltoluene. In a preferred sorbent material, the polyol formed is a diol.

[0063]

[0075] In another embodiment, the polymeric sorbent is prepared by the reaction of a crosslinker with vinyl acetate, which is subsequently modified to form beads containing polyol groups. The reaction can be a copolymerization or a one-pot reaction in which vinyl acetate is added near the end of the initial polymerization and a second free radical polymerization is initiated using unused initiator to add vinyl acetate groups to the surface of the polymer beads. Subsequent modification of the vinyl acetate-containing polymer involves, in order: hydrolysis to convert acetate groups to hydroxyl groups, reaction with epichlorohydrin to form polymer beads containing epoxide groups, and ring-opening to convert the epoxide groups to polyol groups. In a preferred embodiment, the polyol is a diol.

[0064]

[0076] One embodiment of the present invention involves the direct synthesis of polymeric beads containing epoxide groups, followed by ring-opening of the epoxide groups to form polyols. Polymeric beads containing the aforementioned functional groups can be obtained by polymerizing one or more of the following polymerizable vinyl monomers containing epoxide groups in the presence of a crosslinker and a monomer: allyl glycidyl ether, 3,4-dihydroxy-1-butene, 3,4-epoxy-1-butene, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, glycidyl methacrylate, 4-vinyl-1-cyclohexene 1,2-epoxide, and 2-vinyloxirane. Vinyl monomers containing epoxide groups can also be copolymerized with hemocompatible monomers (e.g., NVP, 2-HEMA) to obtain hemocompatible beads containing epoxide groups. In a preferred embodiment, the polyol is a diol.

[0065]

[0077] Yet another embodiment comprises a highly cross-linked polymeric sorbent material containing polyol groups on the surface of the beads, which can be used to treat free radicals or S NThe chemical modification of the sorbent bead surface described above is facilitated by a distinctive feature of highly cross-linked polystyrene; namely, the reactive functional groups of the polymer are primarily present on its surface. Highly cross-linked polystyrene is generally produced by cross-linking polystyrene chains with a large amount of bifunctional compounds, particularly those bearing two reactive chloromethyl groups. The latter alkylates two phenyl groups of adjacent polystyrene chains in a two-step reaction via the Friedel-Crafts reaction, generating two molecules of HCl to form cross-links. During this cross-linking reaction, the three-dimensional network formed acquires rigidity. This characteristic gradually slows down the rate of the second cross-linking reaction; the reduced mobility of the second pendant functional group of the initial cross-linking reagent makes it increasingly difficult to add a second partner suitable for the alkylation reaction. This is particularly characteristic of second functional groups accidentally exposed on the surface of the beads. Therefore, the largest portion (if not the majority) of the pendant unreacted chloromethyl groups in the final highly cross-linked polymer are located on the surface of the beads (or on the surface of the pores). This situation allows the surface of the polymeric beads to be primarily modified by the involvement of said chloromethyl groups in various chemical reactions that allow the attachment of biocompatible and hemocompatible monomers and / or crosslinkers or low molecular weight oligomers. Subsequent introduction of hydroxyl groups followed by reaction with epichlorohydrin produces polymeric sorbents containing epoxide groups on the surface of the beads. These epoxide groups can then be ring-opened to form polyol groups. In one preferred embodiment, the polyol is a diol.

[0066]

[0078] In other embodiments, highly cross-linked polystyrene containing pendant unreacted chloromethyl groups is directly modified in the presence of one or more of the following reagents to form sorbent polymer beads containing a polyol on the surface of the beads (or on the surface of the pores): (±)-3-amino-1,2-propanediol, glycerol, and other polyols. In preferred embodiments, the polyol is a diol.

[0067]

[0079] In yet another embodiment, the surface coating biocompatible and hemocompatible agent poly(vinyl alcohol) also serves as the polyol functional group.

[0080] In certain other embodiments, the sorbent material includes a crosslinked polymeric material derived from the reaction of a crosslinker with one or more of the following polymerizable monomers, followed by a polymerizable zwitterionic monomer in the presence of a free radical initiator: acrylonitrile, allyl glycidyl ether, butyl acrylate, butyl methacrylate, cetyl acrylate, cetyl methacrylate, 3,4-dihydroxy-1-butene, dipentaerythritol diacrylate, dipentaerythritol dimethacrylate, dipentaerythritol tetraacrylate, dipentaerythritol tetramethacrylate, dipentaerythritol triacrylate, dipentaerythritol trimethacrylate, divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone, 3,4-epoxy-1-butene, 1,2-epoxy-9- Decene, 1,2-epoxy-5-hexene, ethyl acrylate, ethyl methacrylate, ethylstyrene, ethylvinylbenzene, glycidyl methacrylate, methyl acrylate, methyl methacrylate, octyl acrylate, octyl methacrylate, pentaerythritol diacrylate, pentaerythritol dimethacrylate, pentaerythritol tetraacrylate, pentaerythritol tetramethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, styrene, trimethylolpropane diacrylate, trimethylolpropane dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, trivinylbenzene, trivinylcyclohexane, vinyl acetate, vinylbenzyl alcohol, 4-vinyl-1-cyclohexene 1,2-epoxide, vinylformamide, vinylnaphthalene, 2-vinyloxirane, and vinyltoluene.Zwitterionic monomers that are polymerizable include one or more of the following: 2-acrylamido-2-methyl-1-propanesulfonic acid sodium salt, [3-(acryloylamino)propyl]-trimethylammonium chloride, 3-[[2-(acryloyloxy)ethyl]-dimethylammonio]-propionate, [2-(acryloyloxy)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide, 2-acryloyloxyethyl phosphorylcholine, [3-(methacryloylamino)propyl]-trimethylammonium chloride, 3-[[2-(methacryloyloxy)ethyl]-dimethylammonio]-propionate, [2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide, and 2-methacryloyloxyethyl phosphorylcholine.

[0068]

[0081] In one embodiment, the polymers of the present invention are prepared by free radical initiated suspension polymerization in a formulated aqueous phase in the presence of an aqueous phase dispersant selected to impart a biocompatible and hemocompatible exterior surface to the formed polymer beads. In some embodiments, the beads are rendered porous by macroreticular synthesis using an appropriately selected porogen (pore former) and a time-temperature profile for polymerization to generate the proper pore structure.

[0069]

[0082] In other embodiments, polymers prepared by suspension polymerization can be made biocompatible and hemocompatible by further grafting biocompatible and hemocompatible monomers or low-molecular-weight oligomers. It has been shown that radical polymerization does not consume all of the vinyl groups of DVB introduced into the copolymer. On average, approximately 30% of the DVB species are not used as crosslinks and remain involved in the network via only one of the two vinyl groups. Therefore, the presence of a relatively large amount of pendant vinyl groups is a distinctive feature of this adsorbent. These pendant vinyl groups can be expected to be preferably exposed on the surface of the polymer beads, and their macropores (if present) should be readily accessible for chemical modification. Chemical modification of the DVB-copolymer surface relies on chemical reactions of the surface-exposed pendant vinyl groups, with the aim of converting these groups into more hydrophilic functional groups. This conversion by free-radical grafting of monomers and / or crosslinkers or low-molecular-weight oligomers provides the first hydrophobic adsorbent with hemocompatible properties.

[0070]

[0083] In yet another embodiment, the radical polymerization initiator is first added to the dispersed organic phase, rather than to the aqueous dispersion medium, as is common in suspension polymerization. During polymerization, many growing polymer chains present their chain-end radicals at the phase interface, which can initiate polymerization in the dispersion medium. Furthermore, radical initiators such as benzoyl peroxide generate radicals relatively slowly. This initiator is only partially consumed during bead formation, even after several hours of polymerization. This initiator readily migrates toward the surface of the beads, activating the pendant vinyl groups of the divinylbenzene moieties exposed on the surface of the beads, thus initiating the graft polymerization of other monomers added after the reaction has proceeded for a period of time. Thus, free radical grafting can occur during the migration of monomer droplets into the polymer beads, thereby incorporating monomers and / or crosslinkers or low-molecular-weight oligomers that impart biocompatibility or hemocompatibility as a surface coating.

[0071]

[0084] Hemoperfusion and perfusion devices consist of a bead bed of polymeric beads packed into a flow-through vessel with either retention screens at both the inlet and outlet ends to retain the bead bed within the vessel or a subsequent retention screen to collect the beads after mixing. Hemoperfusion and perfusion operations are performed by passing whole blood, plasma, or physiological fluid through the packed bead bed. Upon perfusion through the bead bed, toxic molecules are retained by sorption, tortuous path, and / or pore entrapment, while the remaining fluid and intact cellular components pass through essentially unchanged in concentration.

[0072]

[0085] In certain other embodiments, the in-line filter consists of a packed bead bed of polymeric beads in a flow-through vessel with retention screens at both the outlet and inlet ends to retain the bead bed inside the vessel. During a single gravity pass through the packed bead bed, toxic molecules are retained by sorption, tortuous path, and / or pore entrapment, while the remaining fluid and intact cellular components pass through essentially unchanged in concentration.

[0073]

[0086] Certain polymers useful in the present invention (either by themselves or after further modification) are macroporous polymers made from the polymerizable monomers styrene, divinylbenzene, ethylvinylbenzene, and acrylate and methacrylate monomers, such as those listed below by manufacturer. Rohm and Haas Company (now part of Dow Chemical Company): Macroporous polymer-based sorbents, such as Amberlite™ XAD-1, Amberlite™ XAD-2, Amberlite™ XAD-4, Amberlite™ XAD-7, Amberlite™ XAD-7HP, Amberlite™ XAD-8, Amberlite™ XAD-16, Amberlite™ XAD-16 HP, Amberlite™ XAD-18, Amberlite™ XAD-200, Amberlite™ XAD-1180, Amberlite™ XAD-2000, Amberlite™ XAD-2005, Amberlite™ XAD-2010, Amberlite™ XAD-761, and Amberlite™ XE-305, and chromatography grade sorbents such as Amberchrom™ CG 71,s,m,c, Amberchrom™ CG 161,s,m,c, Amberchrom™ CG 300,s,m,c, and Amberchrom™ CG 1000,s,m,c. The Dow Chemical Company: Dowex™ Optipore™ L-493, Dowex™ Optipore™ V-493, Dowex™ Optipore™ V-502, Dowex™ Optipore™ L-285, Dowex™ Optipore™ L-323, and Dowex™ Optipore™ V-503.Lanxess (formerly Bayer and Sybron): Lewatit(TM) VPOC 1064 MD PH, Lewatit(TM) VPOC 1163, Lewatit(TM) OC EP 63, Lewatit(TM) S 6328A, Lewatit(TM) OC 1066, and Lewatit(TM) 60 / 150 MIBK. Mitsubishi Chemical Corporation:Diaion(TM) HP 10, Diaion(TM) HP 20, Diaion(TM) HP 21, Diaion(TM) HP 30, Diaion(TM) HP 40, Diaion(TM) HP 50, Diaion(TM) SP70, Diaion(TM) SP 205, Diaion(TM) SP 206, Diaion(TM) SP 207, Diaion(TM) SP 700, Diaion(TM) SP 800, Diaion(TM) SP 825, Diaion(TM) SP 850, Diaion(TM) SP 875, Diaion(TM) HP 1MG, Diaion(TM) HP 2MG, Diaion(TM) CHP 55A, Diaion(TM) CHP 55Y, Diaion(TM) CHP 20A, Diaion(TM) CHP 20Y, Diaion™ CHP 2MGY, Diaion™ CHP 20P, Diaion™ HP 20SS, Diaion™ SP 20SS, Diaion™ SP 207SS. Purolite Company: Purosorb™ AP 250 and Purosorb™ AP 400, and Kaneka Corp. Lixelle beads.

[0074]

[0087] Certain other polymers useful in the present invention (either by themselves or after further modification) are cellulosic porous materials. Such modifications include free radical or S N Two types of chemistries are used to attach polyols or zwitterionic substrates, along with aryl groups or Alternatively, the addition of a lipophilic substrate containing an alkyl group may be included.

[0075]

[0088] A variety of proteins can be adsorbed by the compositions of the present disclosure, some of which proteins and their molecular weights are listed in the table below.

[0076] [Table 1] [Example]

[0077]

[0089] The following examples are offered by way of illustration and not by way of limitation. Example 1: Base sorbent synthesis CY14175 and CY15077

[0090] Reactor Setup: A four-neck glass lid was attached to a 3 L jacketed cylindrical glass reactor using stainless steel flange clamps and a PFTE gasket. The lid was fitted with a PFTE stirrer bearing, RTD adapter, and water-cooled reflux condenser. A stainless steel stirring shaft with five 60° agitators was threaded through the stirrer bearing and inserted into a digital overhead stirrer. The RTD was threaded through a corresponding adapter and connected to a PolyStat circulating heating and cooling unit. Matching tubing was used to connect the reactor jacket inlet and outlet to the appropriate ports on the PolyStat. The unused port on the lid was used for reactor loading and was plugged at all other times.

[0078]

[0091] Polymerization: The compositions of the aqueous and organic phases are shown in Tables I and II below, respectively. Ultrapure water was dispensed in approximately equal portions into two separate Erlenmeyer flasks, each containing a PFTE-coated magnetic stir bar. The degree of hydrolysis was 85.0-89.0 mole percent, and Poly(vinyl alcohol) (PVA), which has a viscosity of 23.0–27.0 cP in a 4% aqueous solution at 20°C, was dispersed in water in a first flask and heated to 80°C on a hot plate with stirring. Salts (see Table 1, MSP, DSP, TSP, and sodium nitrite) were dispersed in water in a second flask and heated to 80°C on a hot plate with stirring. Circulation of heat transfer fluid from the PolyStat through the reactor jacket was initiated, and the fluid temperature was raised to 60°C. Once the PVA and salts were dissolved, both solutions were simultaneously charged into the reactor using a glass funnel. The digital overhead stirrer was turned on and the rpm was set to a value that would produce an appropriate droplet size during the organic phase addition. The temperature of the aqueous phase in the kettle was set to 70°C. The organic phase was prepared by adding benzoyl peroxide (BPO) to divinylbenzene (DVB) in a 2 L Erlenmeyer flask and stirring until completely dissolved. 2,2,4-Trimethylpentane and toluene were added to the flask and shaken to mix thoroughly. When the temperature of the aqueous phase in the reactor reached 70°C, the organic phase was charged to the reactor using a fine-necked glass funnel. The temperature of the reaction volume decreased as the organic phase was added. A temperature program on the PolyStat was initiated, heating the reaction volume from 60°C to 77°C over 30 minutes, from 77°C to 80°C over 30 minutes, holding the temperature at 80°C for 960 minutes, and then cooling to 20°C over 60 minutes.

[0079] [Table 2]

[0080]

[0092] Finishing: The reaction volume level in the reactor was marked. The overhead stirrer was stopped, the remaining liquid was siphoned out of the reactor, and the reactor was filled up to the mark with room temperature ultrapure water. The overhead stirrer was restarted, and the slurry was heated to 70°C as quickly as possible. After 30 minutes, the stirring was stopped, and the remaining liquid was siphoned out. The polymer beads were washed five times in this manner. For the final wash, the slurry temperature was cooled to room temperature. After the final wash, the polymer beads were washed with 99% isopropyl alcohol (IPA) in the same manner. The 99% IPA was siphoned out and replaced with 70% IPA, and the slurry was transferred to a clean 4 L glass container. Unless otherwise stated, polymers were steam stripped in stainless steel tubes for 8 hours, rewetted in 70% IPA, transferred to DI water, sieved to collect only the bead fraction with diameters between 300 and 600 μm, and dried at 100°C until no further weight loss was observed upon drying.

[0081]

[0093] Polymer CY141 measured by nitrogen desorption isotherm and mercury intrusion porosimetry, respectively The cumulative pore volume data for 75 and CY15077 are shown in Tables III and IV, respectively.

[0082] [Table 3-1]

[0083] [Table 3-2]

[0084] Example 2: Polymer-modified CY15129

[0094] Epoxidation: 50.8 g of dried base polymer CY14175 was added to a 1 L jacketed glass reactor equipped with a Teflon-coated agitator and RTD probe. 300 mL of acetic anhydride (99%) was added to the reactor containing the dried base polymer. The mixture was cooled to 5°C with constant stirring at 100 RPM. 30 mL of hydrogen peroxide solution (30% in water) was added over 30 minutes. The reaction temperature was maintained at 10-15°C for 24 hours with stirring at 100 RPM.

[0085]

[0095] Workup: The reaction mixture was washed with acetic acid and then with DI water until the pH of the reaction supernatant was neutral. The polymer was then dried at 80°C until no further loss was observed upon drying. The dried polymer yield was 61.6g.

[0086]

[0096] The above epoxidation procedure should not be scaled up to reaction volumes greater than 1 L. The formation of diacetyl peroxide can occur when the desired intermediate compound, peracetic acid, combines with excess acetic anhydride. Diacetyl peroxide is known to be shock-sensitive explosive (http: / / cen.acs.org / articles / 89 / i2 / Chemical-Safety-Synthesis-Procedure.html). Therefore, it is emphasized that an alternative epoxidation procedure should be used whenever possible.

[0087]

[0097] Ring opening: 20.0 g of dry epoxide-functionalized polymer was added to a 500 mL jacketed glass reactor equipped with a Teflon-coated agitator and an RTD probe. 70 mL of 70% isopropanol (IPA) was charged to the reactor and the mixture was stirred at 100 RPM. 70 mL of 1 M aqueous NaOH was slowly added. The reaction temperature was increased to 70°C and held at 70°C for 24 hours with stirring at 100 RPM.

[0088]

[0098] Workup: The reaction mixture was cooled to room temperature and washed with DI water until the pH of the reaction supernatant was neutral, resulting in a poly(styrene-co-divinylbenzene) resin functionalized with 1,2-diol groups.

[0089]

[0099] The cumulative pore volume data for polymer CY15129 measured by nitrogen desorption isotherm is shown in Table V below. Table VI shows the atomic concentrations for polymer CY15129 measured by XPS. A logarithmic differential pore volume plot for polymer CY15129 is presented in accompanying Figure 1.

[0090]

[0100] Thrombogenicity was measured by uPTT assay, in which materials were compared to a negative control (plasma only), a positive control (glass beads), and reference beads to determine the degree of contact activation activity. In the uPTT assay, the percent change in clot formation over time compared to the reference material was determined and then grouped according to the following: <25% activator of the intrinsic coagulation pathway, 25-49% moderate activator, 50-74% mild activator, 75-100% minimal activator, and >100% non-activator. Polymer CY15129 (97%) was the minimal activator.

[0091] [Table 4]

[0092] Example 3: Polymer-modified CY15154

[0101] 20.05 g of dried base polymer CY15077 was placed in a Teflon-coated The base polymer was added to a 500 mL jacketed glass reactor equipped with an agitator and RTD probe. A 100 mL slurry was prepared in the reactor by rewetting the dried polymer in DI water. 9.00 g of the zwitterionic neutral methacrylate monomer, [(2-methacryloyloxy)ethyl]-dimethyl-3-(sulfopropyl)ammonium hydroxide, and 1.1 g of ammonium persulfate were dissolved in 100 mL of DI water and the solution was added to the reactor containing the base polymer slurry. The mixture was heated to 75°C and held at 75°C for 24 hours with stirring at 100 RPM.

[0093]

[0102] Workup: The reaction mixture was cooled to room temperature and washed with DI water until the pH of the reaction supernatant was neutral, resulting in a sulfobetaine-functionalized poly(styrene-co-divinylbenzene) resin.

[0094]

[0103] The cumulative pore volume data for polymer CY15154 measured by mercury intrusion porosimetry is shown in Table VII below. Table VIII presents the atomic concentrations for polymer CY15154 measured by XPS. A logarithmic differential pore volume plot for polymer CY15154 is presented in accompanying Figure 2.

[0095]

[0104] Thrombogenicity was measured by uPTT assay, in which materials were compared to a negative control (plasma only), a positive control (glass beads), and reference beads to determine the degree of contact activation activity. In the uPTT assay, the percent change in clot formation over time compared to the reference material was determined and then grouped according to the following: <25% activator of the intrinsic coagulation pathway, 25-49% moderate activator, 50-74% mild activator, 75-100% minimal activator, and >100% non-activator. Polymer CY15154 (89%) was the minimal activator.

[0096] [Table 5]

[0097] Example 4: Polymer-modified CY16029

[0105] 200 mL of base polymer CY14175 wetted in DI water was added to Tef The mixture was added to a 1000 mL jacketed glass reactor equipped with a lon-coated agitator and RTD probe. Excess water was removed from the reactor using a vacuum pump and filter tube. 500 mL of 1.0 M sodium hydroxide was added to the reactor. The mixture was heated to 50°C and held at 50°C for 24 hours with stirring at 100 RPM.

[0098]

[0106] Workup: The reaction mixture was cooled to room temperature and washed with DI water until the pH of the reaction supernatant was neutral, resulting in a diol-functionalized poly(styrene-co-divinylbenzene) resin.

[0099] Example 5: Base sorbent synthesis CY15186

[0107] Reactor Setup: A four-neck glass lid was attached to a 1 L jacketed cylindrical glass reactor using stainless steel flange clamps and a PFTE gasket. The lid was fitted with a PFTE stirrer bearing, RTD adapter, and water-cooled reflux condenser. A stainless steel stirring shaft with four 60° agitators was threaded through the stirrer bearing and inserted into a digital overhead stirrer. The RTD was threaded through a corresponding adapter and connected to a PolyStat circulating heating and cooling unit. Matching tubing was used to connect the reactor jacket inlet and outlet to the appropriate ports on the PolyStat. The unused port on the lid was used for reactor loading and was plugged at all other times.

[0100]

[0108] Polymerization: The compositions of the aqueous and organic phases are shown in Tables IX and X below. Ultrapure water was added to an Erlenmeyer flask containing a PFTE-coated magnetic stir bar. Poly(vinyl alcohol) (PVA), with a degree of hydrolysis of 85.0–89.0 mole percent and a viscosity of 23.0–27.0 cP in a 4% aqueous solution at 20°C, was dispersed in the water in the flask and heated to 80°C with stirring on a hot plate. Circulation of heat transfer fluid from a PolyStat through the reactor jacket was initiated, and the fluid temperature was increased to 60°C. Once the PVA was dissolved, the solution was charged into the reactor using a glass funnel. The digital overhead stirrer was turned on and the rpm was set to a value that would produce the appropriate droplet size during the organic phase addition. The temperature of the aqueous phase in the kettle was set to 70°C. The organic phase was prepared by adding benzoyl peroxide (BPO) to divinylbenzene (DVB) and allyl glycidyl ether (AGE) in a 1 L Erlenmeyer flask and stirring until completely dissolved. 2,2,4-Trimethylpentane and toluene were added to the flask and shaken to mix thoroughly. When the temperature of the aqueous phase in the reactor reached 70°C, the organic phase was charged to the reactor using a fine-necked glass funnel. The temperature of the reaction volume decreased as the organic phase was added. A temperature program on the PolyStat was initiated, heating the reaction volume from 60°C to 77°C over 30 minutes, from 77°C to 80°C over 30 minutes, holding the temperature at 80°C for 960 minutes, and then cooling to 20°C over 60 minutes.

[0101] [Table 6]

[0102]

[0109] Workup: The reaction volume level in the reactor was marked. The overhead stirrer was stopped, the remaining liquid was siphoned out of the reactor, and the reactor was filled up to the mark with room temperature ultrapure water. The overhead stirrer was restarted, and the slurry was heated to 70°C as quickly as possible. After 30 minutes, the stirring was stopped, and the remaining liquid was siphoned out. The polymer beads were washed five times in this manner. For the final wash, the slurry temperature was cooled to room temperature. After the final wash, the polymer beads were washed with 99% isopropyl alcohol (IPA) in the same manner. The 99% IPA was siphoned out and replaced with 70% IPA, and the slurry was transferred to a clean 2 L glass container. Unless otherwise stated, polymers were steam stripped in stainless steel tubes for 8 hours, rewetted in 70% IPA, transferred to DI water, sieved to collect only the bead fraction with diameters between 300 and 600 μm, and dried at 100°C until no further weight loss was observed upon drying.

[0103]

[0110] The cumulative pore volume data for polymer CY15186 measured by mercury intrusion porosimetry is shown in Table XI below.

[0104] [Table 7]

[0105] Example 6: Polymer-modified CY16000

[0111] Ring-opening: 100 mL of polymer CY15186 wetted in 70% IPA was added to a 1 L jacketed glass reactor equipped with a Teflon-coated agitator and an RTD probe. 300 mL of 1 M aqueous NaOH was added slowly. The reaction temperature was increased to 80°C and held at 80°C for 24 hours with stirring at 100 RPM.

[0106]

[0112] Workup: The reaction mixture was cooled to room temperature and washed with DI water until the pH of the reaction supernatant was neutral, resulting in a poly(allyl glycidyl ether-co-divinylbenzene) resin functionalized with 1,2-diol groups.

[0107]

[0113] The cumulative pore volume data for polymer CY16000 measured by mercury intrusion porosimetry is shown in Table XII below. Table XIII presents the atomic concentrations for polymer CY16000 measured by XPS. A logarithmic differential pore volume plot for polymer CY16000 is presented in accompanying Figure 3.

[0108]

[0114] Thrombogenicity was measured by uPTT assay, in which materials were compared to a negative control (plasma only), a positive control (glass beads), and reference beads to determine the degree of contact activation activity. In the uPTT assay, the percent change in clot formation over time compared to the reference material was determined and then grouped according to the following: <25% activator of the intrinsic coagulation pathway, 25-49% moderate activator, 50-74% mild activator, 75-100% minimal activator, and >100% non-activator. Polymer CY16000 (84%) was the minimal activator.

[0109] [Table 8-1]

[0110] [Table 8-2]

[0111] Example 7: Base sorbent synthesis CY16207

[0115] Reactor Setup: A four-neck glass lid was attached to a 3 L jacketed cylindrical glass reactor using stainless steel flange clamps and a PFTE gasket. The lid was fitted with a PFTE stirrer bearing, RTD adapter, and water-cooled reflux condenser. A stainless steel stirring shaft with five 60° agitators was threaded through the stirrer bearing and inserted into a digital overhead stirrer. The RTD was threaded through a corresponding adapter and connected to a PolyStat circulating heating and cooling unit. Matching tubing was used to connect the reactor jacket inlet and outlet to the appropriate ports on the PolyStat. The unused port on the lid was used for reactor loading and was plugged at all other times.

[0112]

[0116] Polymerization: The compositions of the aqueous and organic phases are shown in Tables XIV and XV below, respectively. Ultrapure water was dispensed approximately equally into two separate Erlenmeyer flasks, each containing a PFTE-coated magnetic stir bar. Poly(vinyl alcohol) (PVA), with a degree of hydrolysis of 85.0–89.0 mole percent and a viscosity of 23.0–27.0 cP in a 4% aqueous solution at 20°C, was dispersed in water in the first flask and heated to 80°C on a hot plate with stirring. Salts (see Table 1, MSP, DSP, TSP, and sodium nitrite) were dispersed in water in the second flask and heated to 80°C on a hot plate with stirring. Circulation of heat transfer fluid from the PolyStat through the reactor jacket was initiated, and the fluid temperature was raised to 60°C. Once the PVA and salts were dissolved, both solutions were simultaneously charged into the reactor using a glass funnel. The digital overhead stirrer was turned on and the rpm was set to a value that would produce the appropriate droplet size during the organic phase addition. The temperature of the aqueous phase in the kettle was set to 60°C. The organic phase was prepared by adding benzoyl peroxide (BPO) to divinylbenzene (DVB) and vinyl acetate (VA) in a 2 L Erlenmeyer flask and shaking until completely dissolved. 2,2,4-Trimethylpentane and toluene were added to the flask and shaken to mix thoroughly. When the temperature of the aqueous phase in the reactor reached 60°C, the organic phase was charged to the reactor using a narrow-necked glass funnel. The temperature of the reaction volume decreased as the organic phase was added. The temperature program for the PolyStat was started, heating the reaction volume from 50°C to 67°C over 30 minutes, from 67°C to 70°C over 30 minutes, holding the temperature at 70°C for 960 minutes, and then cooling to 20°C over 60 minutes.

[0113] [Table 9]

[0114]

[0117] Finishing: The reaction volume level in the reactor was marked. The overhead stirrer was stopped, the remaining liquid was siphoned off from the reactor, and the reactor was filled to the mark with room-temperature ultrapure water. The overhead stirrer was restarted, and the slurry was heated to 70 °C as quickly as possible. After 30 minutes, the stirring was stopped, and the remaining liquid was siphoned off. The polymer beads were washed five times in this manner. For the final wash, the slurry temperature was cooled to room temperature. After the final wash, the polymer beads were washed with 99% isopropyl alcohol (IPA) in the same manner. The 99% IPA was siphoned off and replaced with 70% IPA, and the slurry was transferred to a clean 4-L glass container. Unless otherwise noted, the polymer was steam-stripped in a stainless steel tube for 8 hours, rewetted in 70% IPA, transferred into DI water, and sieved to collect only the bead fraction with diameters between 300 and 600 μm, then stored in 70% IPA.

[0115] Example 8: Polymer-modified CY16083

[0118] Ring-opening: 100 mL of polymer CY16207 wetted in 70% IPA was added to a 1 L jacketed glass reactor equipped with a Teflon™ coated agitator and an RTD probe. 300 mL of 1 M aqueous NaOH was added slowly. The reaction temperature was increased to 50° C. and held at 50° C. for 24 hours with stirring at 100 RPM.

[0116]

[0119] Workup: The reaction mixture was cooled to room temperature and washed with DI water until the pH of the reaction supernatant was neutral, resulting in a poly(vinyl acetate-co-divinylbenzene) resin functionalized with diol groups.

[0117]

[0120] The cumulative pore volume data for polymer CY16083 measured by mercury intrusion porosimetry is shown in Table XVI below. A log differential pore volume plot for polymer CY16083 is presented in accompanying Figure 4.

[0118] [Table 10]

[0119] Example 9: Pore structure summary and classification

[0121] The pore structure, including pore size, pore size distribution, and surface properties, is crucial for the adsorption properties of porous materials. IUPAC defines pores as micropores. They are classified into pores, mesopores, and macropores; these are terms widely used in adsorption, catalysis, and other areas.

[0120]

[0122] The micropores have a width of less than 2 nm (<20 Å).

[0123] The mesopores have a width of 2 to 50 nm (20 Å to 500 Å).

[0124] Macropores have widths greater than 50 nm (>500 Å).

[0121]

[0125] Under the IUPAC definition, pore size (or pore diameter) is the distance between two opposing walls of a pore and thus represents the diameter of a cylindrical pore or the width of a slit-shaped pore. In addition to the IUPAC classification, the term transport pore (with a pore diameter greater than 250 Å) is used in the field of activated carbon adsorption. For the purposes of this description, the term "large transport pore" refers to pores with a diameter greater than 2,000 Å as a subgroup of macropores; the term "capacity pore" refers to pores with a diameter greater than 100 Å, which can adsorb small and medium-sized biomolecules and proteins (up to 50 kDa); and the term "effective pore" refers to pores with a diameter in the range of 100 to 250 Å, which are a subgroup of mesopores and have been shown to be the most effective pores for adsorption of small and medium-sized proteins.

[0122]

[0126] Mesopores are useful adsorption sites, while macropores are the internal adsorbent sites. Large transport pores provide a migration pathway for large molecules to reach the adsorption site. Large transport pores provide a more efficient migration pathway, which is particularly important for large molecules, such as large proteins, endotoxins, and other large toxic molecules. The present invention discloses methods to create a wide range of pore size distributions, including large transport pores, general macropores, mesopores, and micropores, to meet the specialized needs of adsorption applications. Table XVII summarizes the pore size distributions of example polymers disclosed in this application, including IUPAC classifications and size fractions important for protein and biomolecule adsorption.

[0123] [Table 11]

[0124] Example 10: Endotoxin removal from plasma in a recirculation model

[0127] 20 mL of heat-inactivated citrated human plasma was spiked with 3 EU / mL endotoxin purchased from Associates of Cape Cod (East Falmouth, MA) and then spiked into a plasma reservoir. The endotoxin-containing plasma was mixed on a stir plate for 15 minutes and recirculated through a 1.5 mL polymer column at a flow rate of 2.5 mL / min. Samples were collected from the reservoir using a sterile pipette tip and diluted 1:20 in endotoxin-free water. All diluted samples were tested with the Pierce LAL Endotoxin Assay from Life Technologies, Corp. (Grand Island, NY). Data for endotoxin removal from plasma in a dynamic recirculation model using modified polymers CY15129, CY15154, CY16000, and CY16029 are shown in Figure 5. Figure 6 presents endotoxin removal data for polymer CY15154 and its unmodified precursor polymer CY15077.

[0125] Example 11: Cytokine removal from bovine whole blood in a recirculation model

[0128] The purified protein was dissolved in 300 mL of 3.8% citrated bovine whole blood (Lamphi Plasma was analyzed by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer's instructions (R&D Systems). Data for cytokine removal from whole blood in a dynamic recirculation model using polymer CY15129 are shown in Figure 7.

Claims

1. A biocompatible polymer system comprising at least one polymer, the polymer comprising a diol functional group; the polymer system being capable of adsorbing endotoxin, the adsorption of the endotoxin occurring with hemoperfusion or by enteral or rectal administration of the polymer; the polymer comprises a solid support having a biocompatible hydrogel coating, the coating comprising poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(N-vinylpyrrolidone), poly(vinyl alcohol), a salt of poly(acrylic acid), a salt of poly(methacrylic acid), or a mixture thereof; Biocompatible polymer systems.

2. 10. The biocompatible polymer system of claim 1, wherein the polymer system is also capable of adsorbing toxins and inflammatory mediators having molecular weights from less than 0.5 kDa up to 1,000 kDa.

3. The biocompatible polymer system of claim 2 , wherein the toxins and inflammatory mediators have a molecular weight of less than 0.5 kDa to 60 kDa.

4. 3. The biocompatible polymer system of claim 2, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies.

5. 10. The biocompatible polymer system of claim 1, wherein the polymer system is also capable of adsorbing one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS).

6. 2. The biocompatible polymer system of claim 1, wherein the polymer system is also capable of adsorbing one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, such as lipoteichoic acid (LTA).

7. 10. The biocompatible polymer system of claim 1, wherein the polymer is produced using suspension polymerization, emulsion polymerization, bulk polymerization, or precipitation polymerization.

8. The polymer is prepared by modification of a cellulosic polymer, and the modification is carried out by free radicals or S Ｎ 2. The biocompatible polymer system of claim 1, comprising two types of chemically attached polyol or zwitterionic substrates, and optionally the attachment of lipophilic substrates containing aryl or alkyl groups.

9. 2. The biocompatible polymer system of claim 1, wherein the polymer system has the form of a solid support, which may include, but is not limited to, beads, fibers, monolithic columns, films, membranes, or semipermeable membranes.

10. The biocompatible polymer system of claim 9 , wherein the solid support has a biocompatible hydrogel coating.

11. 2. The biocompatible polymer system of claim 1, wherein the polymer comprises a large number of pores, and the pore structure of the polymer has a total volume of greater than 0.1 cc / g and less than 5.0 cc / g (dry polymer) of pore sizes ranging from 10 Å to 40,000 Å.

12. The biocompatible polymer system of claim 1 , wherein the polymer is non-porous.

13. The biocompatible polymer system of claim 1 , wherein the polymer is a highly crosslinked polymer.

14. The biocompatible polymer system of claim 1 , wherein the polymer is hemocompatible.

15. 10. The biocompatible polymer system of claim 1, wherein the agent used to impart biocompatibility is either (i) heparin or (ii) a heparin-mimetic polymer.

16. 10. The biocompatible polymer system of claim 1, wherein the polymer is formed and then modified to become biocompatible.

17. 17. The modification that confers biocompatibility of claim 16, wherein the agent used to confer biocompatibility is either (i) heparin or (ii) a heparin-mimetic polymer.

18. A device for removing endotoxins from physiological fluids, comprising a biocompatible polymer system according to any one of claims 1 to 17.

19. 20. The device of claim 18, wherein the device also removes toxins and inflammatory mediators having a molecular weight of less than 0.5 kDa up to 1,000 kDa.

20. 20. The device of claim 19, wherein the toxins and inflammatory mediators have a molecular weight of less than 0.5 kDa to 60 kDa.

21. 20. The device of claim 19, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies.

22. 20. The device of claim 18, wherein the device also removes one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS).

23. 20. The device of claim 18, wherein the device also removes one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, such as lipoteichoic acid (LTA).

24. A device for removing endotoxins from non-physiological fluids, comprising a biocompatible polymer system according to any one of claims 1 to 17, The device, wherein the non-physiological fluid is selected from a laboratory or manufacturing fluid or an aqueous system in one or more of a healthcare facility, a home healthcare application, a medical facility, a biotechnology facility, a biological manufacturing process, a cell culture manufacturing process, and a laboratory.

25. 25. The device of claim 24, wherein the device also removes toxins and inflammatory mediators having a molecular weight of less than 0.5 kDa up to 1,000 kDa.

26. 26. The device of claim 25, wherein the toxins and inflammatory mediators have a molecular weight of less than 0.5 kDa to 60 kDa.

27. 26. The device of claim 25, wherein the toxins and inflammatory mediators comprise one or more of cytokines, pathogen-associated molecular pattern molecules (PAMPs), damage-associated molecular pattern molecules (DAMPs), superantigens, monokines, chemokines, interferons, proteases, enzymes, peptides including bradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin, growth factors, glycoproteins, prions, toxins, bacterial and viral toxins, drugs, vasoactive substances, foreign antigens, and antibodies.

28. 25. The device of claim 24, wherein the device also removes one or more of gram-negative bacteria, gram-negative bacterial fragments, and gram-negative bacterial components, such as lipopolysaccharide (LPS).

29. 25. The device of claim 24, wherein the device also removes one or more of gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components, such as lipoteichoic acid (LTA).

30. The biocompatible polymer system according to any one of claims 1 to 17, in a device suitable for holding the polymer and suitable for incorporation into an extracorporeal circuit.

31. A method of perfusion comprising passing a physiological fluid through a device comprising a biocompatible polymer system according to any one of claims 1 to 17, one or more times, by means of a suitable extracorporeal circuit.

32. 18. The biocompatible polymer system according to any one of claims 1 to 17, contained in a container suitable for holding the polymer for transfusion of a blood product comprising whole blood, packed red blood cells, platelets, albumin, plasma or any combination thereof.

33. The biocompatible polymer system according to any one of claims 1 to 17, for removing endotoxins from blood products including whole blood, plasma or serum, or from other physiological fluids.

34. The biocompatible polymer system according to any one of claims 1 to 17, wherein the polymer is administered enterally or rectally.

35. A polymer system comprising at least one polymer, the polymer comprising a diol functional group; the polymer system being capable of adsorbing endotoxin, the adsorption of the endotoxin occurring with hemoperfusion or by enteral or rectal administration of the polymer; the polymer comprises a solid support having a biocompatible hydrogel coating, the coating comprising poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropyl methacrylate), poly(N-vinylpyrrolidone), poly(vinyl alcohol), a salt of poly(acrylic acid), a salt of poly(methacrylic acid), or a mixture thereof; Polymer system.

36. 36. The polymer system of claim 35, wherein the polymer system is also capable of adsorbing one or more of toxins and inflammatory mediators having a molecular weight of less than 0.5 kDa to 1,000 kDa, gram-negative bacteria, gram-negative bacterial fragments, gram-negative bacterial components such as lipopolysaccharide (LPS), gram-positive bacteria, gram-positive bacterial fragments, and gram-positive bacterial components such as lipoteichoic acid (LTA).

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