[go: up one dir, main page]

WO2018007827A1 - Glycopolymères sulfatés - Google Patents

Glycopolymères sulfatés Download PDF

Info

Publication number
WO2018007827A1
WO2018007827A1 PCT/GB2017/052003 GB2017052003W WO2018007827A1 WO 2018007827 A1 WO2018007827 A1 WO 2018007827A1 GB 2017052003 W GB2017052003 W GB 2017052003W WO 2018007827 A1 WO2018007827 A1 WO 2018007827A1
Authority
WO
WIPO (PCT)
Prior art keywords
sulfated
glycopolymer
glycopolymers
groups
galactose
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2017/052003
Other languages
English (en)
Inventor
Luisa MARTINEZ-POMARES
Francesca MASTROTTO
Giuseppe Mantovani
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Nottingham
Original Assignee
University of Nottingham
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Nottingham filed Critical University of Nottingham
Publication of WO2018007827A1 publication Critical patent/WO2018007827A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/74—Synthetic polymeric materials
    • A61K31/795—Polymers containing sulfur
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0036—Galactans; Derivatives thereof

Definitions

  • the present invention relates to novel sulfated glycopolymers and to the use of these glycopolymers.
  • the novel sulfated glycopolymers may be used in the treatment and/or modulation of inflammation, or for the treatment and/or prevention of ischaemia, including treatment and/or prevention of ischaemia reperfusion injury or ischaemic stroke.
  • Mannose receptor a 175kDa type 1 transmembrane receptor, also referred to as CD206 (Martinez-Pomares, L., 2012).
  • the mannose receptor is expressed in selected populations of key immune cells such as macrophages and other cells (for example, Kaposi's sarcoma spindle cells, dendritic cells and lymphatic endothelial cells and kidney mesangial cells).
  • CD206 consists of the following 5 domains: an amino terminal cysteine-rich region (CR domain), a fibronectin type II repeat (FNII), a series of eight tandem C-type lectin-like carbohydrate recognition domains, a transmembrane domain and an intracellular carboxy -terminal tail.
  • CR domain amino terminal cysteine-rich region
  • FNII fibronectin type II repeat
  • CD206 consists of the following 5 domains: an amino terminal cysteine-rich region (CR domain), a fibronectin type II repeat (FNII), a series of eight tandem C-type lectin-like carbohydrate recognition domains, a transmembrane domain and an intracellular carboxy -terminal tail.
  • FNII fibronectin type II repeat
  • CD206 has been associated with a range of diseases and conditions. For example, previous studies have shown a role of CD206 in the development of kidney crescentic glomerulonephritis in mice whilst other studies have shown that CD206 is overexpressed on alveolar macrophages in lungs of patients with severe chronic obstructive pulmonary disease and suggested its involvement in the pathogenesis of this condition.
  • the present invention uses novel sulfated glycopolymers specific for CD206 which selectively bind to the CR domain and modulate, or even suppress, the endocytic activity of selected CD206 expressing cells.
  • the invention provides a sulfated glycopolymer comprising a polymeric backbone wherein at least one of the monomeric units which form the backbone have a pendant galactose and/or N-acetyl galactosamine group, wherein one or more of the pendant galactose groups are sulfated at one or more of positions 2, 3, 4 and 6 and/or one or more of the N-acetyl galactosamine groups are sulfated at one or more of positions 3, 4 and 6.
  • one or more of the pendant galactose and/or N-acetyl galactosamine groups are sulfated at position 3 and/or 4. In another embodiment, one or more of the pendant N-acetyl galactosamine groups are sulfated at position 3 and/or 6. The one or more of the pendant galactose and/or N-acetyl galactosamine groups may be sulfated at only one position, preferably at position 3 or 4.
  • the sulfated glycopolymer may comprise further pendant groups, these groups may include non- sulfated galactose and/or N-acetyl galactosamine groups or other groups.
  • Further pendant groups may include fluorescent molecules such as fluorescein, rhodamine, coumarin and Bodipy analogues; residues that can react with amine (such as activated (e.g. N-hydroxysuccinimide and pentafluorophenol) esters; residues that can react with thiol such as maleimide, pyridine disulphide, and Michael acceptors e.g. vinyl sulfone, derivatives; hydrophobic groups such as aromatic and aliphatic groups; or other hydrophilic molecules such as glycerol, other sugars like glucose, and poly(ethylene glycol).
  • fluorescent molecules such as fluorescein, rhodamine, coumarin and Bodipy analogues
  • residues that can react with amine such as activated (e.g. N-hydroxysuccinimide and pentafluorophenol) esters
  • residues that can react with thiol such as maleimide, pyridine disulphide, and Michael acceptors e
  • the pendant groups are sulfated galactose and/or N-acetyl galactosamine groups.
  • the invention provides a sulfated glycopolymer consisting of a polymeric backbone wherein at least one of the monomeric units which form the backbone have a pendant galactose and/or N-acetyl galactosamine group, wherein one or more of the pendant galactose groups are sulfated at one or more of positions 2, 3, 4 and 6 and/or one or more of the N-acetyl galactosamine groups are sulfated at one or more of positions 3, 4 and 6.
  • Sulfated galactose or sulfated N-acetyl galactosamine groups which are sulfated at positions 3 or 4 may also be referred to as (S0 4 -3/4)-Gal and (S0 4 -3/4)-GalNAc sulfated groups respectively.
  • At least about 20% of the monomeric units in the glycopolymer have a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • At least about 30% of the monomeric units in the glycopolymer have a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • At least 40%, 50%, 60%, 70%, 80%, 90% or more of the monomeric units have a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • At least about 20% of the monomeric units in the glycopolymer have a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • at least about 30% of the monomeric units in the glycopolymer have a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • at least 40%, 50%, 60%, 70%, 80%, 90% or more of the monomeric units have a pendant galactose and/or N- acetyl galactosamine group which are sulfated at position 3 and/or 4.
  • the invention provides a glycopolymer wherein at least 20 monomeric units carry a pendant galactose and/or N-acetyl galactosamine group wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • At least 30 monomeric units carry a pendant galactose and/or N- acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3 , 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • the glycopolymer may contain 40, 50, 60, 70, 80, 90, 100 or more monomeric units which carry a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • the invention provides a glycopolymer wherein at least 20 monomeric units carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4. More preferably, at least 30 monomeric units carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • the glycopolymer may contain 40, 50, 60, 70, 80, 90, 100 or more monomeric units which carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • the monomeric units carrying a sulfated galactose and/or sulfated N-acetyl galactosamine group may be located consecutively in the glycopolymer.
  • the glycopolymer includes a region wherein at least about 20% of the monomeric units in that region carry a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • the glycopolymer includes a region wherein at least about 30% of the monomeric units in that region carry a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • a region of the glycopolymer has at least 40%, 50%, 60%, 70%, 80%, 90% or more of the monomeric units carrying a pendant galactose and/or N-acetyl galactosamine groups, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetyl galactosamine group is sulfated at one or more of positions 3, 4 and 6.
  • the glycopolymer includes a region wherein at least about 20% of the monomeric units in that region carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4. In another embodiment the glycopolymer includes a region wherein at least about 30% of the monomeric units in that region carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • a region of the glycopolymer has at least 40%, 50%, 60%, 70%, 80%, 90% or more of the monomeric units carrying a pendant galactose and/or N-acetyl galactosamine groups which are sulfated at position 3 and/or 4.
  • the invention provides a glycopolymer which includes a region wherein at least 20 of the monomeric units in that region carry a pendant galactose which is sulfated at one or more of positions 2, 3, 4 and 6 and/or a pendant N-acetyl galactosamine group which is sulfated at one or more of positions 3, 4 and 6, preferably the region comprises at least 30, 40, 50, 60, 70, 80, 90, 100 or more monomeric units which carry a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at position 2, 3 , 4 and/or 6 and the N-acetyl galactosamine group is sulfated at position 3, 4 or 6.
  • the invention provides a glycopolymer which includes a region wherein at least 20 of the monomeric units in that region carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4, preferably the region comprises at least 30, 40, 50, 60, 70, 80, 90, 100 or more monomeric units which carry a pendant galactose and/or N-acetyl galactosamine group which is sulfated at position 3 and/or 4.
  • the region may comprise at least 20 consecutive monomer units, preferably at least 30, 40, 50, 60, 70, 80, 90, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 200 or more monomeric units.
  • the region comprises between about 60 and about 180 consecutive monomeric units, preferably between about 80 and about 180 consecutive monomeric units.
  • the glycopolymer may comprise sufficient pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group is sulfated at one or more of positions 2, 3, 4 and 6 and the N-acetylgalactosamine group is sulfated at one or more of positions 2, 4 and 6 so that the polymer binds to CD206 with an affinity sufficient to inhibit the activity of CD206.
  • the glycopolymer may comprise sufficient pendant galactose and/or N-acetyl galactosamine group which are sulfated at position 3 or 4 so that the polymer binds to CD206 with an affinity sufficient to inhibit the activity of CD206.
  • the activity inhibited is inhibited by at least 10%, more preferably by at least about 25 %, 50% or more.
  • the activity inhibited may be the ability of CD206 to be recycled, which may reduce cell uptake of CD206 ligands and could have consequences in the biological activity of the cells such as response to stimulation and migration.
  • the inhibition is reversible, minimising potential adverse effects of CD206 inhibition.
  • the inhibition may last for at least 12 hours, or for at least 24 hours or for at least 36 hours or more.
  • the inhibition may last no more than 36 hours, no more than 24 hours or no more than 12 hours.
  • the sulfated glycopolymer comprises a polymeric backbone wherein at least 90% of the monomeric units which form the backbone have a pendant galactose and/or N-acetyl galactosamine group, wherein the pendant galactose group and/or N-acetyl galactosamine group is sulfated at position 3 and/or 4.
  • the percentage of pendant groups in the glycopolymer can be determined by 3 ⁇ 4 NMR.
  • the polymeric backbone may comprise at least 20 repeating monomeric units, preferably at least 30, 40, 50, 60, 70, 80, 90, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 200, or more wherein reference to repeating monomeric units means a monomeric unit of the backbone, for example, acrylamide or methacrylate unit.
  • the backbone may comprise 30- 100 repeating monomeric units.
  • the glycopolymer may also comprise galactose and/or N-acetyl galactosamine groups which are not sulfated.
  • the polymeric backbone may comprise any suitable monomeric units, this may include carbohydrates such as a sugar (polysaccharide), poly(vinyl monomer) including poly(meth)acrylate and poly(meth)acrylamide, polyester, polycarbonate, polyurethane, and polyamides.
  • the polymeric backbone may include blocks which do not include carbohydrate pendant units, such as PEG-glycopolymers and other block copolymers.
  • the sulfated glycopolymer may be at least about 50kDa, at least about 60kDa, at least about 70kDa, at least about 80kDa, at least about 90kDa.
  • the sulfated glycopolymer may be less than about l OOkDa, less than about 90kDa, less than about 80kDa, less than about 70kDa, or less than about 60kDa.
  • the sulfated glycopolymer may have an M n of between about 1 and about 50kDa, or between about 5 and about 25kDa. In some embodiments, the sulfated glycopolymer may have a M n of at least 60kDa.
  • Sulfated glycopolymers having a smaller M n may be appropriate for instances where it is intended that the sulfated glycopolymer will cross the blood-brain barrier.
  • the M n of sulfated glycopolymers according to the invention may be calculated using any standard method, this may include by determining the number of repeat units by l NMR or by using size exclusion chromatography (SEC).
  • the pendant sulfated galactose and/or sulfated N-acetyl galactosamine groups are attached to the monomeric units of the polymeric backbone via a linker.
  • a linker in used, all pendant groups may be attached using the same linker type, or more than one type of linker may be used.
  • the linker is a glycoside linker. In some embodiments, the linker is an amino-terminated linker. In some embodiments, the linker includes a 1 ,2,3-triazole group. In some embodiments, the linkers may comprise - 0(CH 2 )3 S(CH 2 ) 2 NH-. In some embodiments, the linker may be a chain of from 1 to 20 member atoms selected from carbon, oxygen, sulfur, nitrogen and phosphorus. The linker may be a straight chain or branched.
  • the pendant sulfated galactose and/or sulfated N-acetyl galactosamine groups may be attached to the backbone of the glycopolymer via a glycosylic linker connected to the anomeric carbon (C I) of the galactose and/or N- acetyl galactosamine molecule.
  • the sulfated glycopolymer of the invention may specifically bind to CD206 also known as the mannose receptor. Avidity for CD206 can be estimated by surface plasmon resonance (SPR) analysis, to provide K D values in the micromolar-nanomolar range.
  • the sulfated glycopolymer may be specific for the CR domain of the CD206 receptor.
  • the sulfated glycopolymers of the invention may prevent or delay mannose receptor recycling.
  • Sulfated glycopolymers of the invention may be used to treat, prevent, alleviate, ameliorate, relieve, delay onset of, inhibit progression of, reduce severity of, and/or reduced incidence of one or more symptoms or features of ischaemia reperfusion injury (IRI), acute kidney injury (AKI), or ischaemic stroke.
  • IRI ischaemia reperfusion injury
  • AKI acute kidney injury
  • ischaemic stroke ischaemia reperfusion injury
  • treat treatment
  • treating means the management and care of a subject for the purpose of combating a condition, such as a disease or a disorder.
  • the term is intended to include the full spectrum of treatments for a given condition from which the subject is suffering, such as administration of the sulfated glycopolymer to alleviate the symptoms or complications, to delay the progression of the disease, disorder or condition, to alleviate or relief the symptoms and complications, and/or to cure or eliminate the disease, disorder or condition as well as to prevent the condition, wherein prevention is to be understood as the management and care of a subject for the purpose of combating the disease, condition, or disorder and includes the administration of the mannose receptor (CD206) modulators to prevent the onset of the symptoms or complications.
  • the subject to be treated is preferably a mammal, in particular a human, but it may also include animals, such as dogs, cats, horses, cows, sheep and pigs.
  • IRI Ischaemia reperfusion injury
  • ATI Acute Tubular Injury
  • IRI is of particular concern after transplantation or acute ischaemic episodes in a range of organs including the kidney, liver and heart. Whilst a range of potential treatments have been investigated in preclinical and clinical trials, including drugs such as cyclosporine and hydrogen sulphide, and strategies based on hyperthermia or stem cell infusions, to date, no consistent clinically acceptable means of attenuating IRI is available. Thus devising a means to attenuate or minimise IRI represents a significant unmet clinical need that could produce substantial healthcare and economic benefits across many diverse medical specialities.
  • the invention provides the use of sulfated glycopolymers according to the invention in the manufacture of a medicament for the treatment and/or prevention of IRI, acute kidney injury, myocardial ischaemia, ischaemic stroke, cancer or autoimmune disease.
  • the invention provides sulfated glycopolymers according to the invention for use in the treatment of IRI, acute kidney injury, myocardial ischaemia, ischaemic stroke, cancer or autoimmune disease.
  • the invention provides a method of treating IRI, acute kidney injury, myocardial ischaemia, ischaemic stroke, cancer or autoimmune disease comprising administering to a subject in need thereof a therapeutically effective amount of a sulfated glycopolymer according to the invention.
  • the invention provides the use of sulfated glycopolymers according to the invention in the manufacture of a medicament for the treatment of a macrophage-related or mannose-binding C-type lectin receptor high expressing cell related disease.
  • the invention provides sulfated glycopolymers according to the invention for use in the treatment of a macrophage-related or mannose-binding C-type lectin receptor high expressing cell related disease .
  • the invention provides a method of treating a macrophage-related or a mannose-binding C-type lectin receptor high expressing cell related disease comprising administering to a subject in need thereof a therapeutically effective amount of a sulfated glycopolymer according to the invention.
  • Macrophage-related and other mannose-binding C-type lectin receptor high expressing cell-related diseases for which the glycopolymers and/or compositions herein may be used include, but are not limited to: acquired immune deficiency syndrome (AIDS), acute disseminated encephalomyelitis (AD EM), Addison's disease, agammaglobulinemia, allergic diseases, alopecia areata, Alzheimer's disease, amyotrophic lateral sclerosis, ankylosing spondylitis, antiphospholipid syndrome, antisynthetase syndrome, arterial plaque disorder, asthma, atherosclerosis, atopic allergy, atopic dermatitis, autoimmune aplastic anemia, autoimmune cardiomyopathy, autoimmune enteropathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune hypothyroidism, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral neuropathy, autoimmune pancreatitis, autoimmune polyendocrine syndrome,
  • the sulfated glycopolymer may be used to treat arthritis including enthesitis-related arthritis, inflammatory arthritis, rheumatoid arthritis inflammatory bowel disease, inflammatory dementia, interstitial cystitis, interstitial pneumonitis, juvenile idiopathic arthritis (aka juvenile rheumatoid arthritis), psoriatic arthritis; cancer; cardiac conditions; ischaemia reperfusion injury; stroke; transplantation; infection; and autoimmunity.
  • a sulfated glycopolymer according to the invention may be used in organ transplantation.
  • the sulfated glycopolymer may be used to perfuse the organ prior to transplantation, this may minimise activation of resident macrophages.
  • the sulfated glycopolymer may be used to treat the organ recipient, it may modulate activation of macrophages recruited to the transplanted organ.
  • the transplanted organ may be a heart or liver.
  • the invention provides the use of sulfated glycopolymers according to the invention in the manufacture of a medicament for targeting tumor- associated macrophages, such as for the treatment of cancer.
  • the invention provides sulfated glycopolymers according to the invention for use in targeting tumor-associated macrophages, such as for the treatment of cancer.
  • the invention provides a method of treating a cancer comprising administering to a subject in need thereof a therapeutically effective amount of a sulfated glycopolymer according to the invention.
  • the invention also provides a pharmaceutical composition comprising a sulfated glycopolymer according to the invention and a pharmaceutically acceptable excipient.
  • compositions to be used comprise a therapeutically effective amount of a sulfated glycopolymer according to the invention together with one or more pharmaceutically acceptable excipients, such as carriers, diluents, fillers, disintegrants, lubricating agents, binders, colorants, pigments, stabilizers, preservatives, antioxidants, and/or solubility enhancers.
  • pharmaceutically acceptable excipients such as carriers, diluents, fillers, disintegrants, lubricating agents, binders, colorants, pigments, stabilizers, preservatives, antioxidants, and/or solubility enhancers.
  • the pharmaceutical compositions can be formulated by techniques known in the art, such as the techniques published in Remington's Pharmaceutical Sciences, 20th Edition.
  • the glycopolymers and pharmaceutical compositions of the invention may be formulated as for oral, parenteral, such as intramuscular, intravenous, subcutaneous, intradermal, intraarterial, intracardial, intracavity, intraperitoneal, transdermal, rectal, nasal, topical, aerosol or vaginal administration.
  • the pharmaceutical composition may be formulated as a dosage form for oral administration.
  • the disclosed glycopolymers or compounds can be administered via any suitable method.
  • the disclosed glycopolymers or compounds can be administered parenterally into the parenchyma or into the circulation so that the disclosed glycopolymers or compounds reach target tissues.
  • the disclosed glycopolymers or compounds can be administered directly into or adjacent to a tumour mass.
  • a physician will determine the actual dosage which will be most suitable for an individual subject.
  • the specific dose level and frequency of dosage for any particular individual subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual subject undergoing therapy.
  • the sulfated glycopolymer according to the present invention may be administered systemically, i.e. intravenously.
  • the sulfated glycopolymer according to the present invention may be administered directly to the organ and/or systemically. It will be appreciated that optional features applicable to one aspect or embodiment of the invention can be used in any combination, and in any number. Moreover, they can also be used with any other aspects or embodiments of the invention in any combination and in any number. This includes, but is not limited to, the dependent claims from any claim being used as dependent claims for any other claims of this application.
  • Figures 1A-E - demonstrates the structure and ligand-binding modalities of the mannose receptor/CD206 and the structure of the glycopolymers of the present invention.
  • Figure 1A - shows a schematic representation of the CD206 receptor.
  • CR Cysteine-rich domain which binds 3/4-O-sulfate galactose (S0 4 -3/4)-Gal) and
  • N-(acetyl galactosamine) (S0 4 -3/4)-GalNAc); FNII: collagen-binding Fibronectin type II domain; CTLD: C-type lectin-like domain which binds mannose (Man), fucose (Fuc), N-(acetyl glucosamine) (GlcNAc).
  • Figure IB - shows the chemical structure, composition, and molecular weight of sulfated glycopolymers ( 1 st generation).
  • M n (kDa) is calculated from the number of repeating units as estimated by : H NMR, while D was obtained by SEC analysis. All glycopolymers were fluorescently labelled with 2,4,5,7,7'- pentafluorofluorescein (Oregon Green, l ex/em 488/530 nm) to enable detection in in vitro assays.
  • Figure 1C - shows details of the polymers specifically tested with human monocyte-derived macrophages.
  • Figure ID - demonstrates that sulfated polymers on the invention bind to the CR domain of the mouse mannose receptor (moMR-CR or CD206-CR) .
  • Hydrophobic S0 4 -3 -Gal sulfated glycopolymers known to interact specifically with the CR domain (2 ⁇ g/mL) were immobilized on wells of a 96 well ELISA plate .
  • Figure IE - demonstrates that sulfated polymers on the invention, exemplified by S 100-DP I 87 bind to the CR domain of the human mannose receptor (huMR-CR).
  • MAXISORP 96-well plates were coated with polymers (5 ⁇ g/ml in PBS) and incubated overnight at 4 °C. Wells were washed 3x with TBS buffer and huCR-Fc chimeric protein consisting of the CR-domain of human MR fused to the Fc portion of mouse IgG (Martinez-Pomares et al., 2005) was added at different concentrations (0.2, 1 , 5, 10 ⁇ g/ml in TBS) and incubated for 2 hours.
  • Washing step was repeated 3x with TBS buffer.
  • To detect binding anti-mouse IgG Fc-specific antibody (alkaline phosphatase conjugated and diluted 1 : 1000) was added and incubated for 1 hour.
  • Wells were washed again 3x with TBS buffer and 2x with AP buffer.
  • Alkaline phosphatase activity was detected using SIGMAFAST p-nitrophenyl phosphate diluted in AP buffer.
  • OD405 was measured in a Multiskan FC (Thermo Scientific) . Data was processed in Excel and Graphpad Prism 6.
  • Figures 2A-D - demonstrates the internalisation of sulfated glycopolymers by CD206-presenting cells.
  • Figure 2A - shows flow cytometry analysis of the uptake of 3-O-sulfo- galactosylated glycopolymers (GPs) with different sugars epitope densities.
  • CD206 + -CHO and CD206 -CHO cells were incubated with the glycopolymers ( 1.0 ⁇ g mL "1 ) for 30 minutes at 37°C. After washing, cells were collected and analysed by FACS . The data presented shows a reduced uptake with increasing density of S0 4 -3-Gal by CHO-MR cells.
  • Figure 2B - shows confocal microscopy analysis of CD206 + -CHO following incubation with Oregon Green (OG) tagged glycopolymers ( 100 ⁇ g mL "1 ) for 1 h at 37°C.
  • Cells were stained with HOECHST (H) for nuclei and imaged under bright field (BF) and fluorescence (FL) mode.
  • OG Oregon Green
  • FL fluorescence
  • Figure 2C - shows the uptake of glycopolymers by WT and CD206-KO macrophages quantified by FACS after incubation with 1.0 ⁇ g mL "1 glycopolymers solutions for 30 minutes at 37°C.
  • Figure 2D - shows time-dependent uptake of glycopolymers ( 1.0 ⁇ g mL "1 ) by CD206 + -CHO cells. After treatment at various incubation times, cells were washed and uptake of Oregon Green-tagged glycopolymers was quantified by FACS .
  • Figures 3A-C - demonstrate the mechanism of modulation of CD206 endocytic activity in vitro, and CD206 activity inhibition in vivo.
  • Figure 3A - shows the quantification of CD206 receptor at the cell surface.
  • Cells were incubated for 120 min with 480 ⁇ (S0 4 -3-Galioo%) 32 glycopolymers (concentration refers to individual sugar repeating units), or cell medium only (positive control cells), then CD206 was immunostained and membrane-bound CD206 quantified by FACS .
  • Data are the means ⁇ SD of duplicates of two independent experiments.
  • Figure 3B - shows the quantification of total cell CD206 (MR). Blot analysis of cell lysates from MR + -CHO cells treated with (S0 4 -3-Galioo%)32 solutions
  • Figure 3C - shows the pH does not affect binding of sulfated glycopolymers to the MR-CR domain.
  • the 7.4-6.0 pH range was chosen to simulate the conditions which GP-CD206 complexes encounter by going from cell membrane to endosomes.
  • Dissociation constant K D values were estimated by surface plasmon resonance (SPR) analysis, using immobilized CD206, in 10 mM HEPES, 5 mM CaCl 2 , 0.005% tween-20, 150 mM NaCI, pH 7.4, 6.5, 6. Higher values indicate more favorable dissociation of GP-CD206 (MR) complexes, which would facilitate the recycling of CD206 receptor to the cell membrane.
  • SPR surface plasmon resonance
  • Figures 4A-E - demonstrate the sulfated glycopolymers of the invention inhibit CD206 endocytic activity in vitro.
  • Figure 4A - shows glycopolymers of the invention can direct cell uptake of gelatine mediated by CD206 in MR + -CHO through its FN II domain.
  • Cells were pre-treated for (A) 30 minutes or (B) 2 hours with a range of glycopolymers with different sugar epitopes and molecular weight (480 ⁇ in sugar binding units). Control cells were incubated over the same period with cell medium. Cells were then incubated for further 2 hours with fluorescently- tagged gelatin (texas red gelatin, 80 ⁇ g mL "1 ) in the presence (co-incubation) or in the absence of glycopolymers.
  • fluorescently- tagged gelatin texas red gelatin, 80 ⁇ g mL "1
  • FIG. 4B - shows glycopolymers of the invention can reduce cell uptake of gelatine mediated by CD206 in human IL-4 treated macrophages.
  • the IL4 treated macrophages (M-CSF+IL-4) display elevated levels of expression of the mannose receptor (MR) compared to the control macrophages (M-CSF).
  • Human CD 14+-monocyte-derived macrophages were grown on low adherence 24-well plates for 7 days in the presence of M-CSF or M-CSF+IL-4. Polymers were added to a final concentration of 48 ⁇ , media with M-CSF (50 ng/ml) and IL-4 (25 ng/ml) was used to give each well a final volume of 1.1ml and incubated at 37 °C for 6 hours. Polymers S 100-DP 187, M 100-DP 187, G100-
  • DP 187, S 100-DP240 and G100-DP240 were tested (see Figure 1 C).
  • Cells were then incubated in the presence of gelatin ( 10 ⁇ g/ml) for 1 hour at 37 °C in total volume of 1.2 ml per well. Plates were placed on ice for 20 minutes for the cells to detach. Macrophages were harvested by gently pipetting, collected into 15 ml Falcon tubes and centrifuged at 350 x g at 4 °C for 5 minutes. The number of cells was adjusted to 200.000 per 100 ⁇ in X-Vivo 15. Cells were transferred to 1.5 ml Eppendorf tubes (200 ⁇ /tube).
  • Fluorescence quantification was done in a MoFlo Astrios (Beckman Coulter) and analysed using Kaluza software. Oregon Green channel (GPs): Laser 488nm, emission spectrum 500-526nm. Texas Red channel (Gelatine): Laser 561nm, emission spectrum 604-624nm.
  • Collagen is a MR-ligand so it is clear these polymers inhibit MR-mediated endocytosis in human macrophages.
  • the effect of the S I 00 polymers is less prominent in control macrophages because these cells also express the collagen receptor Endo l 80 which is downregulated by IL4.
  • Figure 4C - shows inhibition of CD206 endocytic activity elicited by a single initial treatment with sulfated glycopolymers of the invention.
  • MR + -CHO cells were incubated for 2 hours with (A) (SO 4 -3 -Gal l 00%) 32 or (B) (S0 4 -3 - Gal l 00%) i 87 (480 ⁇ in sugar binding units). Control cells were incubated over the same period with cell medium. Cells were then washed and incubated with cell medium. At a scheduled time the medium was replaced with a 80 ⁇ g ml/ 1 of TR-Gelatin solution and incubated for 1 hour.
  • Data is expressed as a percentage of gelatin uptake of untreated CD206 + -CHO control cells (right column of each panel, TR-Gelatin), as quantified by flow cytometry. Data were analyzed by two-way ANOVA with Bonferroni post-tests, *p ⁇ 0.05, * *p ⁇ 0.01.
  • Figure 4D shows fluorescent microscopy of kidney and liver sections from animals injected with glycopolymer (GP) followed by TR-Gelatin and control TR-Gelatin only injected animal.
  • GP glycopolymer
  • TR-Gelatin control TR-Gelatin only injected animal.
  • CD206 is expressed on mesangial cells only.
  • the figures show in the left panel a glomerulus from a mouse injected with sulfated GP followed by TR- Gelatin, demonstrating little (red) gelatin glomerular uptake, in contrast to the galactosylated GP-treated and control mice, in which there is significant TR-Gelatin uptake into the mesangial cells of the glomerulus.
  • liver sections from the same animals show attenuated TR-Gelatin uptake in the sulfated GP-treated animals, in contrast to the other two control animals in which there is more substantial uptake.
  • Figure 4E - shows that in vivo in mice polymers of the invention co-localise with the mannose receptor in at least the liver. Wt mice were injected with OG-labelled polymer S 100-DP 187 (green) and sacrificed 1 h post injection.
  • Liver was collected, processed for immunolabelling for Mannose receptor (MR, grey) and analysed by confocal microscopy.
  • Cell nucleus was labelled with DAPI (blue).
  • the DAPI image shows cells in the liver section by staining the nucleus.
  • MR (CD206) staining shows the location of the mannose receptor in Kupffer cells and liver sinusoidal endothelial cells.
  • S0 4 -3-Gal-DP 187 fluorescence shows where the polymer has bound.
  • the composite image overlays the S0 4 -3-Gal-DP 187 staining with the MR staining and shows the two co-localise.
  • FIG. 5 - shows (SO 4 -3 -Gal l 00%) i 87 reduces Acute Tubular Injury after IRI.
  • A 200 ⁇ , iv, 2 h prior IRI and 24 h after IRI.
  • Figure 6 - demonstrates the internalisation of polymers of the invention by human macrophages. Furthermore the data demonstrates that human macrophages are not killed in the process, that the uptake occurs in a dose dependent manner and that there is more uptake in IL-4-treated macrophages (IL-4 upregulates expression of MR). This data demonstrates that the polymers of the invention may be used to target macrophages and to modulate macrophages.
  • Human CD 14+-monocyte-derived macrophages grown on low adherence 24- well plate for 7 days in the presence of M-CSF or M-CSF+IL-4 were placed on ice for 20 minutes to allow cells to detach. Macrophages were harvested by gently pipetting and collected into 15 ml Falcon tubes and centrifuged at 350 x g, 4 °C for 5 minutes. The number of cells was adjusted to 100.000 per 200 ⁇ in X-Vivo 15 (Lonza). Cells were transferred to 1.5 ml Eppendorf tubes (200 ⁇ /tube) and 200 ⁇ of polymers (S 100-DP 187, M 100-DP 187 and G100-DP 187, see Figure 1 C) were added to each tube .
  • the tubes were incubated at 37 °C for one hour. After incubation, cells were washed with 400 ⁇ of X-Vivo 15 and centrifuged at 1000 rpm (model Prism R), 4 °C, for 5 minutes. The pellet was washed in 400 ⁇ of ice-cold PBS and centrifuged at 1000 rpm, at 4 °C for 5 minutes. 200 ⁇ of PBS was added to each tube and the content was transferred to FACS tubes containing 200 ⁇ of 1 % formaldehyde in PBS . Fluorescence quantification was done in FC 500 (Beckman Coulter) and analysed using Kaluza software.
  • Figure 7 - demonstrates that the internalisation, as previously shown in Figure 6, is increased in the presence of IL4, which increases the expression of the mannose receptor, and is temperature dependent. That is, a temperature of 37 ° C is needed for the internalisation of the polymers, demonstrating active uptake process.
  • the data demonstrates that the polymers are able to target intracellular compartments of the macrophages and thus are able to change their phenotype and modulate inflammation.
  • the tubes were incubated at 37 °C or 4 °C for one hour. After incubation, cells were washed with 400 ⁇ of X-Vivo 15 and centrifuged at 1000 rpm (model Prism R), 4 °C, for 5 minutes. The pellet was washed in 400 ⁇ of ice-cold PBS and centrifuged at 1000 rpm, at 4 °C for 5 minutes. 200 ⁇ of PBS was added to each tube and the content was transferred to FACS tubes containing 200 ⁇ of 1 % formaldehyde in PBS . Fluorescence quantification was done in FC 500 (Beckman Coulter) and analysed using Kaluza software.
  • Figure 8 - demonstrates that both 1 st and 2 nd generation sulfated polymers of the invention act on the same receptor, as demonstrated by unlabelled S I 00- DP240 inhibiting uptake of OG-labelled S 100-DP I 87 by human macrophages.
  • Human CD 14+-monocyte-derived macrophages grown on low adherence 24- well plate for 7 days in the presence of M-CSF were pre-treated with DP-240 series GPs (2 nd generation, S 100-DP240 or G100-DP240; 20 ⁇ , 40 ⁇ , 80 ⁇ ) for 1 hour and some were left untreated. After one hour pre-treatment the Oregon Green-labelled polymer S 100-DP I 87 ( 1 st generation) was added at 10 ⁇ to the appropriate wells. Samples were incubated for 2 h or 6 h. The plates were placed on ice for 20 min and macrophages were collected in 1.5 ml Eppendorf tubes, centrifuged at 400 x g, 4oC, 5 min.
  • Pellets were washed with lml of ice cold X-Vivo 15 to each tube and centrifuged at 350 x g, 4°C, 5 min. Cells were washed again. Supernatant was poured off and 200 ⁇ cold X-Vivo 15 was added to each tube. Cells were transferred to FACS tubes containing 200 ⁇ of 1 % formaldehyde in PBS . Cells were analysed on FACS500.
  • polymer S 100-DP240 (which is unlabelled) inhibits association of S 100-DP I 87 (which is labelled with Oregon green) with human macrophages. This effect is not seen with the control G100-DP240.
  • Figure 9 - demonstrates the effect of polymers of the invention of macrophage activation.
  • Figure 9A summarises the hypothesis of the involvement of macrophages and IRI.
  • Figure 9B shows the effect of polymers of the invention in an in vitro system intended to mimic macrophage activation.
  • the data presented shows that in the presence of the sulfated polymers the concentration of mannose receptor on the surface of macrophages decreases. Whereas, with a non-sulfated polymer the level of mannose receptor is unchanged.
  • the sulfated polymers are changing the macrophage phenotype by trapping the mannose receptors within the cell.
  • the polymers also change the surface expression levels of CD l ib and MHCII. Normally, in the presence of IFNy macrophages increase the level of MHCII expression on their surface . However, in the presence of the sulfated polymers of the invention no increase in MHCII expression is observed.
  • sulfated polymers of the invention prevent the normal activation of macrophages and that the polymers may be used to modulate macrophages and inflammation.
  • the polymers could be used to treat inflammation driven diseases, perhaps by preventing the activation of M l macrophages, this may be useful in the treatment of IRI.
  • Trimethylsilyl propargyl methacrylate was synthesized starting from 3 -(trimethylsilyl) propargyl alcohol following the method described by Ladmiral, V. et al., Journal of the American Chemical Society 2006, 128 ( 14), 4823-4830. Synthesis of N-(Ethyl)-2-pyridylmethanimide (2)
  • N-(Ethyl)-2-pyridylmethanimide was prepared as described by Haddleton, D.
  • the maleimide-protected initiator was synthesized as previously reported by Mantovani, G. et al, Journal of the American Chemical Society 2005, 127 (9), 2966- 2973.
  • 3-O-sulfo-galactose (7) was synthesized adopting a modified version of the method reported by Manning, D. D., et al , Journal of the American Chemical Society 1997, 119 ( 13), 3 161 -3 162 and Uzawa, H., et al , Chemical Communications 1998, (21), 23 1 1 -23 12.
  • 2 '-azidoethyl-0--D-galactopyranoside 1.00 g, 4.02 mmol
  • MeOH/EtOAc 2 1 ( 15 mL) and phenylboronic acid (490 mg, 4.02 mmol) were added.
  • trimethylsilylpropargylmethacrylate (1) (2.00 g, 10.2 mmol), N-(ethyl)-2-pyridylmethanimine ligand (2) (4.5 mg, 0.34 mmol) and maleimide-protected initiator (3) (30.4 mg, 0.849 mmol) were dissolved in anhydrous toluene (6 mL) and transferred in a Schlenk tube. The tube was sealed with rubber septum and subjected to five freeze-pump-thaw cycles. A second Schlenk tube containing Cu(I)Br (24 mg, 0.17 mmol) was evacuated and filled with nitrogen.
  • the toluene solution was cannulated into the second tube then placed into an oil bath and maintained at 30°C under stirring.
  • the polymerization kinetic was monitored by scheduled withdraw of samples analyzed by : H NMR and GPC.
  • the polymerization was stopped at 70% of conversion by opening the tube to the air.
  • the polymer solution was passed through a short pad of alumina, the column was washed several times with toluene and the solvent removed under reduced pressure .
  • Fluorescent dye and sugars were attached to the polymer backbone by clicking the required components to the poly (propargyl methacrylate) polymeric precursor.
  • the preparation of Oregon green 100% galactose sulfate glycopolymer (S0 4 - 3-Galioo%) 32 is described below.
  • Gelatin-Texas Red conjugate was prepared following the procedure described by Hummert, E., et al , Cellulose 2013, 20 (2), 919-93 1 .
  • a 10 mg mL -1 gelatin solution was prepared by dispersing 100 mg of gelatin in 10 mL of 0. 1 M sodium bicarbonate buffer, pH 9, at RT. The suspension was left under mild stirring for 3 hours . Then the mixture was gently heated under stirring in a water bath at 45 °C, until gelatin complete dissolution.
  • a stock solution of Sulforhodamine 101 acid chloride (Texas Red) was prepared dissolving 2.5 mg of the fluorescent dye in 250 ⁇ . of anhydrous MeCN.
  • 1-thyoglycerol (10.0 g, 92.5 mmol) was dissolved in anhydrous THF (70 mL), the solution was cooled to 10°C in a water bath and a 60% w/w dispersion of NaH in mineral oil (3.7 g, 92 mmol) was added portion-wise. After each addition the suspension was degassed under N 2 for 10 minutes. The reaction mixture was left under stirring for 1 hour, then CS 2 (14.1 g, 185 mmol) was slowly added drop-wise. After 18 hour the organic solvent was removed under reduced pressure and the excess CS 2 was removed under high vacuum. The crude product was re-dissolved in methanol and precipitated in Et 2 0 to obtain 9b as a yellow viscous oil (15 g, 73 mmol, 79%).
  • D-galactosyloxyethyl acrylamide 10 (5.00 g, 18.0 mmol) was dissolved in extra-dry MeOH (30 mL) and phenylboronic acid (2.2 g, 18 mmol) was added to the flask followed by the addition of EtOAc (15 mL).
  • EtOAc 15 mL
  • a Dean-Stark apparatus was attached to the reaction vessel and filled with EtOAc.
  • the reaction mixture was heated to reflux and maintained under stirring for 3 hours.
  • the mixture was then cooled down to RT and the volume was reduced to 10 mL.
  • a fresh portion of EtOAc (15 mL) was added together with dibutyltyn oxide (5.66 g, 19.9 mmol) and the mixture was heated to reflux for further 2 hours under stirring.
  • 3-O-sulfo-D-galactosyloxyethyl acrylamide (1.65 g, 3.97 mmol) was placed in a Schlenk tube equipped with a magnetic bar and dissolved in 1.52 mL of milliQ water. 64.7 ⁇ ⁇ (6.47 mg, 22.1 ⁇ ) of a freshly prepared 100 mg mL "1 dioxane stock solution of 2,3-dihydroxypropyl (l-oxo-l-(prop-2-yn-l-ylamino)propan-2-yl) carbonotrithioate RAFT Agent CTA (9) were added to the reaction vessel. The tube was sealed with a rubber septum and degassed by bubbling argon at RT for 20 minutes.
  • VA-044 stock solution aliquots prepared as described above, were performed until at least 90% of monomer conversion was reached.
  • the mixture was cooled down to RT and diluted to 10 mL with milliQ water.
  • Ethanolamine 27 mg, 27. 1 ⁇ , 448 ⁇
  • iodoacetic acid 250 mg, 1.34 mmol.
  • the solution was stirred at RT for 18 hours, and then further diluted with water (final volume 15 mL) and dialysed with a 1 kDa MWCO regenerated cellulose membrane against 5 L of DI water for 3 days, with at least 2 water exchange per day.
  • the aqueous solution inside the dialysis bag was then freeze-dried to yield (S0 4 -3-Gal) i8o as white powder (912 mg, 13.4 ⁇ , 61.2% mol/mol).
  • Glycopolymers were fluorescently labelled via the CuAAC click reaction.
  • (Gal) i 80 (630 mg, 13.9 ⁇ ) was dissolved in 8 mL of milliQ water, added of CuS0 4 pentahydrate ( 1.74 mg, 6.97 ⁇ ) and degassed by bubbling nitrogen for 20 minutes.
  • a stock solution of Oregon Green Azide ( 16.8 mg mL "1 ) was prepared in MeOH/Acetone 1 : 1 and 768 ⁇ ⁇ ( 12.9 mg, 27.9 ⁇ ) were added to the polymer solution.
  • a freshly made 10 mg mL "1 stock solution of sodium ascorbate in milliQ water was degassed by bubbling argon for 20 minutes.
  • (S0 4 -3-Gal / 0 ) 3 2 glycopolymer with average DP (degree of polymerisation) 32, and 66 % of polymer 3-O-sulfated galactose (S0 4 -3-Gal) repeating units.
  • the remaining 34% of total repeating units are galactose (Gal) moieties.
  • In (SO ⁇ -Ga ⁇ the remaining 67% of total repeating units are galactose (Gal) moieties.
  • mice were kept under specific, pathogen-free conditions and used at 8- 12 weeks of age . Mice were housed at Queen Medical Centre, Central Animal House, University of Nottingham Medical School. All animals were handled in accordance with institutional guidelines issued by the Home Office, United Kingdom.
  • Chinese Hamster Ovary (CHO and CHO-MR) cell lines were maintained in Dulbecco's modified Eagle 's medium/Ham's F- 12 nutrients (Gibco, Eggenstein, Germany) containing 10% Fetal Bovine Serum (FBS), 2 mM L-glutamine, 100 U mL " penicillin and 100 ⁇ g mL "1 streptomycin, at 37°C, 5% C0 2 and 95 % relative umidity.
  • Stable transductants for CHO-MR were selected using 0.6 mg mL "1 geneticin.
  • Bone marrow derived macrophages were obtained from WT and MR ⁇ ' ⁇ C57BL/6 femurs and tibias bone marrow cells using L929 cell conditioning media (LCCM) or M-CSF as source of granulocyte/macrophage colony stimulating factor.
  • LCCM L929 cell conditioning media
  • M-CSF M-CSF
  • Cells were cultured in 20 cm bacteriologic plastic (BP) petri dishes by re-suspension in 25 mL of R10 media (RMPI supplemented with 10% Fetal bovine serum, 15 % LCCM or 50 ng mL "1 M-CSF, 2 mM L-glutamine, 100 U mL ⁇ penicillin and 100 ⁇ g mL _1 streptomycin).
  • the cells were centrifuged at 1000 rpm for 5 minutes, washed with 40 and 10 mL of opti-MEM® and re-suspended in 10 mL of opti-MEM® at a concentration of 1.5xl 0 6 cells mL "1 for seeding.
  • CHO or CHO-MR cells were seeded in 24 wells/plate (400 ⁇ , 6.25xl 0 5 cells mL "1 ) and grown overnight at 37°C, 5% C0 2 . The medium was then removed, the wells were washed with 2x500 of PBS and incubate for 30 minutes with opti- MEM®; the media was aspirated and low and high molecular weight Oregon Green labeled glycopolymers solutions ( 1 ⁇ g mL "1 , 400 ⁇ ) diluted in opti-MEM® supplemented with 2 mM L-glutamine, 100 U mL ⁇ penicillin and 100 ⁇ g mL " Streptomycin were added to the wells.
  • MFI mean fluorescence intensity
  • CHO and CHO-MR cells were seeded in 24 wells/plate (400 ⁇ , 6.25xl 0 5 cells mL "1 ) and grown overnight at 37°C, 5% C0 2 .
  • CHO and CHO-MR cells were seeded in 24 wells/plate (400 ⁇ , 3.75xl 0 5 cells mL "1 ) and grown overnight at 37°C, 5% C0 2 .
  • the media was then discharged, the wells washed with 2x500 ⁇ , of PBS, and incubated with opti-MEM®. After 30 minutes, the medium was removed and low and high molecular weight 100% sulfated Oregon Green labeled glycopolymers were added to the wells (400 480 ⁇ sugar units concentration diluted in opti-MEM®). After 2 h of incubation at 37°C the cells were washed with 3x lmL PBS, and 400 ⁇ ⁇ of opti-MEM® were added to each well.
  • CHO-MR cells were seeded in 24 wells/plate (400 ⁇ , 6.25xl0 5 cells mL "1 ) and grown overnight at 37°C, 5% C0 2 . The media was then discharged, the wells washed with 2x500 ⁇ ⁇ of PBS, and incubated for 30 minutes with opti-MEM®. The medium was then replaced with low or high molecular weight 100% sulfated Oregon Green labeled glycopolymers (400 ⁇ , 480 ⁇ sugar units concentration, diluted in opti-MEM ® ).
  • Proteins from cell lysates (2.5 ⁇ g protein content) were separated on 6% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels under non-reducing conditions and then transferred to nitrocellulose membrane using a Trans Blot® Cell apparatus overnight at 200 mA. After blocking with blocking buffer (0.1% tween 20, 5% low fat milk in PBS) for 1 h, membranes were incubated with anti-MR mAb (MR5D3, 2 ⁇ g mL _1 in blocking buffer) for 1 h.
  • blocking buffer (0.1% tween 20, 5% low fat milk in PBS
  • Membranes were washed 3 times with 0.1% tween 20 in PBS and MR was detected using an HRP- conjugated goat-anti-Rat IgG (1 : 1000 dilution in blocking buffer, 1 h at RT). Bound Abs were visualized by enhanced chemiluminescence reagent (ECL system, Amersham Pharmacia Biotech, Bucks, UK) and recorded on a film.
  • ECL system Amersham Pharmacia Biotech, Bucks, UK
  • CHO-MR cells were seeded in 12 wells/plate ( 1 mL/well, 650xl 0 3 cells mL "1 ) and grown overnight at 37°C, 5% C0 2 . The media was then discharged, the wells washed with 2x500 of PBS, and incubated with opti-MEM®. After 30 minutes the medium was replaced with low or high molecular weight sulfated Oregon Green labeled glycopolymers ( 1.00 mL/well, 480 ⁇ sugar units concentration diluted in opti-MEM®).
  • CHO-MR cells were allowed to adhere to glass slide (pre-treated with HC1 1 N and extensively rinsed with RPMI) overnight. Cells were subsequently incubated for 1 h at 37°C with 100 mg mL "1 solutions of 100% sulfated glycopolymers, diluted in opti-MEM®. After gentle washing with PBS (3x lmL) cells were fixed with 4% paraformaldehyde in PBS for 10 minutes at RT. Cells were washed as described above and nuclei were stained with 1 ⁇ g/mL HOECHST solution in PBS for 15 minutes at RT.
  • coverslips were mounted with DAKO fluorescent mounting medium and viewed by confocal microscopy using a Zeiss LSM 700 microscope and ZEN201 1 software for images elaboration. Unstained control samples or samples treated with 100% galactosylated glycopolymers were analyzed to check for cell autofluorescence (data not shown).
  • a 63x oil objective was used for acquiring all images. Section images in the z- dimension were collected.
  • Binding experiments were performed by Surface Plasmon Resonance (SPR) on a BIAcore 3000. Approximately 1300-2000 response units (RU) sMR were immobilized on a CM-5 sensor chip surface by amine coupling, which according to formula ( 1 ) would result in an R max of approximately 100 RU during kinetic binding experiments:
  • R max analyte MW x immobilized ligand level (1)
  • C57BL/6 mice underwent a unilateral renal ischaemia reperfusion injury (IRI), by clamping the left renal artery for 35 minutes, followed by 48-hour recovery.
  • IRI renal ischaemia reperfusion injury
  • Glycopolymers were injected according to different regimens which included one intravenous injection prior to IRI, one IV injection before and one afterwards and a three dose protocol with one injection prior to IRI, 2 hours later and 24 hours later. All injections were through the tail vein.
  • Urine was collected 12 hours before sacrifice, which was performed after 48 hours.
  • the ischaemic left kidney and control right kidney were scored on histology using a tubular injury score, assessing four variables, tubular dilatation, cast formation, tubular necrosis and loss of tubular brush border (each scored from 0-5).
  • Urine was used for biomarker studies including assessment of KIM- 1 and NGAL. A significant decrease in tubular injury scores was observed in ischaemic kidneys from animals treated with the sulfated but galactosylated controlglycopolymers , with the greatest inhibition observed using the three dose regimen.
  • CD206 binding modalities and glycosylated multivalent ligands
  • Figure 1A shows the structure of the CD206 endocytic receptor, which has three distinct extracellular binding domains: / ' .
  • C Lectin-Type Domain (CLTD) which recognizes Man, Fuc, and GluNAc carbohydrates in a Ca 2+ -dependent manner, / ' / ' . collagen-binding Fibronectin type II (FN II) domain, and / ' / ' / ' .
  • Cysteine-Rich (CR) lectin domain which recognizes (S0 4 -3/4)-Gal and (S0 4 -3/4)-GalNAc sulfated sugars.
  • a tyrosine-based motif in the cytoplasmic intracellular tail directs the delivery of mannosylated ligands to early endosomes (Figure 1A).
  • Two families of multivalent glycoligands for selective targeting of the different lectin- type domains of CD206 were prepared as shown in Figure I B. Macroligands displaying D-galactopyranose-3 -O-sulfate (S0 4 -3 -Gal) repeating units were expected to provide selective binding to CR domain. All polymers had a mucin-like structure, with the sugar binding units grafted to the polymer backbone through a glycoside linker, leaving groups at C3 and C4 of the sugar rings, available for lectin binding.
  • the avidity of binding of carbohydrate-containing polymers to lectin receptors often depends on the polymer chain length, which also directly affects the ability of these multivalent ligands to span over multiple copies of receptors at cell membranes.
  • CD206 mediates clathrin-dependent endocytosis of CD206 ligands to endosomal compartments. Once there the ligand-receptor complexes dissociate and CD206 recycles back to the cell membrane as shown in Figure IB. At every given time most of CD206 is intracellular, with recycling turnover typically occurring in ca. 20 min. Results suggested that after initial cell uptake S0 4 -3 -Gal multivalent ligands prevented further CD206-mediated endocytosis.
  • S0 -3-Gal multivalent ligands were found to bind very strongly to CD206 and induce a significant decrease in the amount of receptor at the cell membrane.
  • the strength of binding of glycopolymers to CD206 was investigated under a range of conditions which simulated the change in environmental conditions which occurs when the ligands bind the receptor at the cell membrane and are subsequently internalised into relatively acidic endosomal compartments.
  • SPR analysis showed that the avidity of binding of S0 -3-Gal GPs for CD206 slightly decreased from pH 7.4, typical of extracellular environment, to more acidic pH (6.5 and 6.0), with dissociation constants K D in the ⁇ range (Figure 3D). Modulation of CD206 (MR) endocytic activity in vitro and in vivo.
  • CD206 + -CHO cells were chosen as they are more robust than WT macrophages, which facilitated prolonged time-course experiments to be carried out.
  • CD206 + -CHO were pre-treated with either (S0 4 -3-Galioo%) 32 or (S0 -3 - Galioo%) i87 ( 15 ⁇ in sugar binding units) for 30 or 120 min, then Gelatin-TR was added to a final concentration of 80 ⁇ g ml/ 1 . After 2 hours of co-incubation, cell uptake was quantified by FACS . Data clearly showed that internalisation of Gelatin- TR was significantly reduced as compared with untreated cell incubated with Gelatin- TR for 2 h (positive control) .
  • CD206 + -CHO pre-incubated with analogous non CD206- binding Gal-glycopolymers gave a Gelatin-TR uptake similar to that observed with untreated cells (as shown in Figure 5A).
  • the uptake patterns did not significantly change by increasing the duration of polymer pre-incubation from 30 to 120 min. Similar trends were also observed when after the initial pre-incubation glycopolymers were removed from the culture media, and cells were treated with Gelatin-TR in the absence of glycopolymers, suggesting that after initial polymer pre-incubation, sustained inhibition of CD206-mediated endocytosis could be attained even in the absence of sulfated glycopolymers.
  • the duration of the inhibition of CD206 endocytic activity was investigated via time course inhibition experiments.
  • lectins In addition to CD206 (MR), a range of other lectins are known to recognise mannose- rich molecular patterns in vivo - e.g. Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin (DC-SIGN, CD209) and Mannose-Binding Lectin (MBL), which can activate the lectin pathway of the complement system. Conversely, lectins able to selectively recognise S0 4 -3-Gal motifs with sufficient avidity are much less known. Targeting the CR domain therefore offers a potent route for selective binding of CD206 in vivo.
  • DC-SIGN Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin
  • MBL Mannose-Binding Lectin
  • the data demonstrates how the endocytic activity of the CD206 mannose receptor can be modulated both in vitro and in vivo by selectively targeting its distinct lectin-type domains with appropriately designed synthetic glycopolymers according to the invention.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des glycopolymères sulfatés comprenant un squelette polymère, au moins l'une des unités monomères qui forment le squelette possédant un groupe pendant galactose et/ou un groupe N-acétyl galactosamine, un ou plusieurs des groupes pendants galactose étant sulfatés en une ou plusieurs des positions 2, 3, 4 et 6 et/ou un ou plusieurs des groupes N-acétyl galactosamine étant sulfatés en une ou plusieurs des positions 3, 4 et 6. L'invention concerne également l'utilisation de glycopolymères sulfatés dans le traitement de maladies, par exemple les lésions d'ischémie-reperfusion, l'atteinte rénale aigüe, l'ischémie myocardique, l'accident vasculaire cérébral ischémique, le cancer ou les maladies auto-immunes.
PCT/GB2017/052003 2016-07-07 2017-07-07 Glycopolymères sulfatés Ceased WO2018007827A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1611849.9A GB201611849D0 (en) 2016-07-07 2016-07-07 Sulfated glycopolymers
GB1611849.9 2016-07-07

Publications (1)

Publication Number Publication Date
WO2018007827A1 true WO2018007827A1 (fr) 2018-01-11

Family

ID=56890934

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2017/052003 Ceased WO2018007827A1 (fr) 2016-07-07 2017-07-07 Glycopolymères sulfatés

Country Status (2)

Country Link
GB (1) GB201611849D0 (fr)
WO (1) WO2018007827A1 (fr)

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108530570A (zh) * 2018-02-08 2018-09-14 中国海洋大学 聚降冰片烯骨架糖聚合物的制备方法及其在岩藻聚糖硫酸酯模拟物合成中的应用
US11038771B2 (en) 2019-04-26 2021-06-15 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing a metadata driven rules engine on blockchain using distributed ledger technology (DLT)
US11144335B2 (en) 2020-01-30 2021-10-12 Salesforce.Com, Inc. System or method to display blockchain information with centralized information in a tenant interface on a multi-tenant platform
US11244313B2 (en) 2019-01-31 2022-02-08 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing declarative smart actions for coins and assets transacted onto a blockchain using distributed ledger technology (DLT)
US11257073B2 (en) 2018-01-31 2022-02-22 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing machine learning models for smart contracts using distributed ledger technologies in a cloud based computing environment
CN114207117A (zh) * 2019-04-12 2022-03-18 激流生物科学有限公司 用于调节巨噬细胞活性的方法
US11288280B2 (en) 2018-10-31 2022-03-29 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing consumer data validation, matching, and merging across tenants with optional verification prompts utilizing blockchain
US11431693B2 (en) 2018-01-31 2022-08-30 Salesforce.Com, Inc. Systems, methods, and apparatuses for seeding community sidechains with consent written onto a blockchain interfaced with a cloud based computing environment
US11488176B2 (en) 2019-01-31 2022-11-01 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing certificates of authenticity of digital twins transacted onto a blockchain using distributed ledger technology (DLT)
US11568437B2 (en) 2018-10-31 2023-01-31 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing commerce rewards across tenants for commerce cloud customers utilizing blockchain
US11611560B2 (en) 2020-01-31 2023-03-21 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing consensus on read via a consensus on write smart contract trigger for a distributed ledger technology (DLT) platform
US11783024B2 (en) 2019-01-31 2023-10-10 Salesforce, Inc. Systems, methods, and apparatuses for protecting consumer data privacy using solid, blockchain and IPFS integration
US11803537B2 (en) 2019-01-31 2023-10-31 Salesforce, Inc. Systems, methods, and apparatuses for implementing an SQL query and filter mechanism for blockchain stored data using distributed ledger technology (DLT)
US11811769B2 (en) 2019-01-31 2023-11-07 Salesforce, Inc. Systems, methods, and apparatuses for implementing a declarative, metadata driven, cryptographically verifiable multi-network (multi-tenant) shared ledger
US11824970B2 (en) 2020-01-20 2023-11-21 Salesforce, Inc. Systems, methods, and apparatuses for implementing user access controls in a metadata driven blockchain operating via distributed ledger technology (DLT) using granular access objects and ALFA/XACML visibility rules
US11824864B2 (en) 2019-01-31 2023-11-21 Salesforce, Inc. Systems, methods, and apparatuses for implementing a declarative and metadata driven blockchain platform using distributed ledger technology (DLT)
US11875400B2 (en) 2019-01-31 2024-01-16 Salesforce, Inc. Systems, methods, and apparatuses for dynamically assigning nodes to a group within blockchains based on transaction type and node intelligence using distributed ledger technology (DLT)
US11876910B2 (en) 2019-01-31 2024-01-16 Salesforce, Inc. Systems, methods, and apparatuses for implementing a multi tenant blockchain platform for managing Einstein platform decisions using distributed ledger technology (DLT)
US11880349B2 (en) 2019-04-30 2024-01-23 Salesforce, Inc. System or method to query or search a metadata driven distributed ledger or blockchain
US11886421B2 (en) 2019-01-31 2024-01-30 Salesforce, Inc. Systems, methods, and apparatuses for distributing a metadata driven application to customers and non-customers of a host organization using distributed ledger technology (DLT)
US11899817B2 (en) 2019-01-31 2024-02-13 Salesforce, Inc. Systems, methods, and apparatuses for storing PII information via a metadata driven blockchain using distributed and decentralized storage for sensitive user information
US11971874B2 (en) 2019-01-31 2024-04-30 Salesforce, Inc. Systems, methods, and apparatuses for implementing efficient storage and validation of data and metadata within a blockchain using distributed ledger technology (DLT)
US11995647B2 (en) 2019-04-30 2024-05-28 Salesforce, Inc. System and method of providing interoperable distributed and decentralized ledgers using consensus on consensus and delegated consensus
US12344594B2 (en) 2019-12-09 2025-07-01 Howard Hughes Medical Institute Red-shifted fluorophores
WO2025147580A1 (fr) * 2024-01-05 2025-07-10 Isp Investments Llc Copolymères séquencés comprenant des unités de répétition et obtenus par polymérisation de diester cyclique et de monomères acryliques, et leurs applications
US12493862B2 (en) 2018-01-31 2025-12-09 Salesforce, Inc. Systems, methods, and apparatuses for implementing smart flow contracts using distributed ledger technologies in a cloud based computing environment
US12526155B2 (en) 2022-06-06 2026-01-13 Salesforce, Inc. Multi-signature wallets in public trust ledger actions via a database system

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5821312A (en) * 1996-09-20 1998-10-13 Geltex Pharmaceuticals, Inc. Acid-functionalized saccharides as polyvalent anti-infectives
WO2000078821A1 (fr) * 1999-06-17 2000-12-28 Wisconsin Alumni Research Foundation Procedes et reactifs formant des jeux ordonnes polyvalents et bibliotheques combinatoires de jeux ordonnes polyvalents
WO2007104948A2 (fr) * 2006-03-10 2007-09-20 Warwick Effect Polymers Ltd. Polymères

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5821312A (en) * 1996-09-20 1998-10-13 Geltex Pharmaceuticals, Inc. Acid-functionalized saccharides as polyvalent anti-infectives
WO2000078821A1 (fr) * 1999-06-17 2000-12-28 Wisconsin Alumni Research Foundation Procedes et reactifs formant des jeux ordonnes polyvalents et bibliotheques combinatoires de jeux ordonnes polyvalents
WO2007104948A2 (fr) * 2006-03-10 2007-09-20 Warwick Effect Polymers Ltd. Polymères

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
CHERNYAK, A. Y. ET AL., CARBOHYDRATE RESEARCH, vol. 223, no. 0, 1992, pages 303 - 309
GORDON E J ET AL: "Synthesis of end-labeled multivalent ligands for exploring cell-surface-receptor-ligand interactions", CHEMISTRY AND BIOLOGY, CURRENT BIOLOGY, LONDON, GB, no. 7, 10 December 1999 (1999-12-10), pages 9 - 16, XP002318565, ISSN: 1074-5521 *
GRONBSEK H ET AL., J HEPATOL, 2016, Retrieved from the Internet <URL:http://dx.doi.org/10.1016/j.jhep.2015.11.021>
HADDLETON, D. M. ET AL., MACROMOLECULES, vol. 32, no. 7, 1999, pages 2110 - 2119
HUMMERT, E. ET AL., CELLULOSE, vol. 20, no. 2, 2013, pages 919 - 931
LADMIRAL, V. ET AL., JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 128, no. 14, 2006, pages 4823 - 4830
LU SU ET AL: "Glycocalyx-Mimicking Nanoparticles for Stimulation and Polarization of Macrophages via Specific Interactions", SMALL, vol. 11, no. 33, 1 September 2015 (2015-09-01), DE, pages 4191 - 4200, XP055406228, ISSN: 1613-6810, DOI: 10.1002/smll.201403838 *
MANNING, D. D. ET AL., JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 119, no. 13, 1997, pages 3161 - 3162
MANTOVANI, G. ET AL., JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 127, no. 9, 2005, pages 2966 - 2973
SUN, W.-C. ET AL., THE JOURNAL OF ORGANIC CHEMISTRY, vol. 62, no. 19, 1997, pages 6469 - 6475
UZAWA, H. ET AL., CHEMICAL COMMUNICATIONS, no. 21, 1998, pages 2311 - 2312
YAMAGUCHI S ET AL: "Inhibition of PrP^S^c formation by synthetic O-sulfated glycopyranosides and their polymers", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ELSEVIER, AMSTERDAM, NL, vol. 349, no. 2, 20 October 2006 (2006-10-20), pages 485 - 491, XP024924462, ISSN: 0006-291X, [retrieved on 20061020], DOI: 10.1016/J.BBRC.2006.08.072 *
YOSHIHIRO NISHIDA ET AL: "A Facile Synthetic Approach to L- and P-Selectin Blockers via Copolymerization of Vinyl Monomers Constructing the Key Carbohydrate Modules of Sialyl Lewis X Mimics", BIOMACROMOLECULES, vol. 1, no. 1, 1 March 2000 (2000-03-01), pages 68 - 74, XP055406119, ISSN: 1525-7797, DOI: 10.1021/bm990011o *

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12493862B2 (en) 2018-01-31 2025-12-09 Salesforce, Inc. Systems, methods, and apparatuses for implementing smart flow contracts using distributed ledger technologies in a cloud based computing environment
US11257073B2 (en) 2018-01-31 2022-02-22 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing machine learning models for smart contracts using distributed ledger technologies in a cloud based computing environment
US11431693B2 (en) 2018-01-31 2022-08-30 Salesforce.Com, Inc. Systems, methods, and apparatuses for seeding community sidechains with consent written onto a blockchain interfaced with a cloud based computing environment
CN108530570B (zh) * 2018-02-08 2020-03-27 中国海洋大学 聚降冰片烯骨架糖聚合物的制备方法及其在岩藻聚糖硫酸酯模拟物合成中的应用
CN108530570A (zh) * 2018-02-08 2018-09-14 中国海洋大学 聚降冰片烯骨架糖聚合物的制备方法及其在岩藻聚糖硫酸酯模拟物合成中的应用
US11288280B2 (en) 2018-10-31 2022-03-29 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing consumer data validation, matching, and merging across tenants with optional verification prompts utilizing blockchain
US11568437B2 (en) 2018-10-31 2023-01-31 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing commerce rewards across tenants for commerce cloud customers utilizing blockchain
US11811769B2 (en) 2019-01-31 2023-11-07 Salesforce, Inc. Systems, methods, and apparatuses for implementing a declarative, metadata driven, cryptographically verifiable multi-network (multi-tenant) shared ledger
US11876910B2 (en) 2019-01-31 2024-01-16 Salesforce, Inc. Systems, methods, and apparatuses for implementing a multi tenant blockchain platform for managing Einstein platform decisions using distributed ledger technology (DLT)
US11488176B2 (en) 2019-01-31 2022-11-01 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing certificates of authenticity of digital twins transacted onto a blockchain using distributed ledger technology (DLT)
US11244313B2 (en) 2019-01-31 2022-02-08 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing declarative smart actions for coins and assets transacted onto a blockchain using distributed ledger technology (DLT)
US11971874B2 (en) 2019-01-31 2024-04-30 Salesforce, Inc. Systems, methods, and apparatuses for implementing efficient storage and validation of data and metadata within a blockchain using distributed ledger technology (DLT)
US11783024B2 (en) 2019-01-31 2023-10-10 Salesforce, Inc. Systems, methods, and apparatuses for protecting consumer data privacy using solid, blockchain and IPFS integration
US11803537B2 (en) 2019-01-31 2023-10-31 Salesforce, Inc. Systems, methods, and apparatuses for implementing an SQL query and filter mechanism for blockchain stored data using distributed ledger technology (DLT)
US11899817B2 (en) 2019-01-31 2024-02-13 Salesforce, Inc. Systems, methods, and apparatuses for storing PII information via a metadata driven blockchain using distributed and decentralized storage for sensitive user information
US11886421B2 (en) 2019-01-31 2024-01-30 Salesforce, Inc. Systems, methods, and apparatuses for distributing a metadata driven application to customers and non-customers of a host organization using distributed ledger technology (DLT)
US11824864B2 (en) 2019-01-31 2023-11-21 Salesforce, Inc. Systems, methods, and apparatuses for implementing a declarative and metadata driven blockchain platform using distributed ledger technology (DLT)
US11875400B2 (en) 2019-01-31 2024-01-16 Salesforce, Inc. Systems, methods, and apparatuses for dynamically assigning nodes to a group within blockchains based on transaction type and node intelligence using distributed ledger technology (DLT)
CN114207117A (zh) * 2019-04-12 2022-03-18 激流生物科学有限公司 用于调节巨噬细胞活性的方法
US11038771B2 (en) 2019-04-26 2021-06-15 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing a metadata driven rules engine on blockchain using distributed ledger technology (DLT)
US11880349B2 (en) 2019-04-30 2024-01-23 Salesforce, Inc. System or method to query or search a metadata driven distributed ledger or blockchain
US11995647B2 (en) 2019-04-30 2024-05-28 Salesforce, Inc. System and method of providing interoperable distributed and decentralized ledgers using consensus on consensus and delegated consensus
US12344594B2 (en) 2019-12-09 2025-07-01 Howard Hughes Medical Institute Red-shifted fluorophores
US11824970B2 (en) 2020-01-20 2023-11-21 Salesforce, Inc. Systems, methods, and apparatuses for implementing user access controls in a metadata driven blockchain operating via distributed ledger technology (DLT) using granular access objects and ALFA/XACML visibility rules
US11144335B2 (en) 2020-01-30 2021-10-12 Salesforce.Com, Inc. System or method to display blockchain information with centralized information in a tenant interface on a multi-tenant platform
US11611560B2 (en) 2020-01-31 2023-03-21 Salesforce.Com, Inc. Systems, methods, and apparatuses for implementing consensus on read via a consensus on write smart contract trigger for a distributed ledger technology (DLT) platform
US12526155B2 (en) 2022-06-06 2026-01-13 Salesforce, Inc. Multi-signature wallets in public trust ledger actions via a database system
WO2025147580A1 (fr) * 2024-01-05 2025-07-10 Isp Investments Llc Copolymères séquencés comprenant des unités de répétition et obtenus par polymérisation de diester cyclique et de monomères acryliques, et leurs applications

Also Published As

Publication number Publication date
GB201611849D0 (en) 2016-08-24

Similar Documents

Publication Publication Date Title
WO2018007827A1 (fr) Glycopolymères sulfatés
Bhattacharya et al. Sulfation modulates the targeting properties of hyaluronic acid to P-selectin and CD44
JP7177131B2 (ja) アポトーシスを誘導するための組成物及び方法
EP3142684A2 (fr) Conjugués de ligands peptidiques se liant à icam/vcam et à la sélectine
Zhang et al. ROS-triggered nanoinducer based on dermatan sulfate enhances immunogenic cell death in melanoma
Tang et al. TGF-β blocking combined with photothermal therapy promote tumor targeted migration and long-term antitumor activity of CAR-T cells
Zaiden et al. Inhibition of CD44v3 and CD44v6 function blocks tumor invasion and metastatic colonization
Vongchan et al. N, N, N-Trimethyl chitosan nanoparticles for the delivery of monoclonal antibodies against hepatocellular carcinoma cells
WO2022187631A1 (fr) Hydrogels thermosensibles libérant des micelles utilisés en tant que système d&#39;administration thérapeutique
EP2946793A1 (fr) Cellules tumorales de ciblage de particules de silicium
EP3793610A1 (fr) Conjugués comprenant un polymère sensible à la température et un ligand capable de se lier à l&#39;ergostérol
ES2684969T3 (es) Inductor de tolerancia inmune
US12433857B2 (en) Conjugate and cancer therapeutic agent
Yang et al. Redox-sensitive targeted nanoparticles synthesized from heparin mimetics for potential glioma treatment application
JP7454263B2 (ja) 高分子薬剤
IL322871A (en) Compositions and methods for targeting tumor-associated macrophages
JP7587583B2 (ja) 新たな免疫賦活剤及び免疫療法におけるその使用
JP7555827B2 (ja) アントラサイクリン系化合物を含むミセル剤と免疫賦活剤との組み合わせ医薬
Chen et al. Supramolecular self-assembly of multifunctional carrageenan-based Nanomicelles for effective tumor therapy via apoptosis and immunogenic cell death
JP2025519467A (ja) 化学的に修飾されたヘパリン
US20170067012A1 (en) Hyaluronic acid derivatives and composition for cell-surface engineering using the same
CZ2020137A3 (cs) Glykopolymer, způsob jeho přípravy a jeho použití jako léčivo
Guțoaia Chitosan Nanoplexes For Target siRNA Delivery
CZ2020133A3 (cs) Glykopolymer, způsob jeho přípravy a jeho použití jako léčivo
CZ2019572A3 (cs) Glykopolymer, způsob jeho přípravy a jeho použití jako léčivo

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17745857

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17745857

Country of ref document: EP

Kind code of ref document: A1