WO2021016924A1 - 可用于治疗银屑病的阳离子聚合物及颗粒 - Google Patents

可用于治疗银屑病的阳离子聚合物及颗粒 Download PDF

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WO2021016924A1
WO2021016924A1 PCT/CN2019/098604 CN2019098604W WO2021016924A1 WO 2021016924 A1 WO2021016924 A1 WO 2021016924A1 CN 2019098604 W CN2019098604 W CN 2019098604W WO 2021016924 A1 WO2021016924 A1 WO 2021016924A1
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polyquaternium
polymer
cationic
segment
copolymer
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PCT/CN2019/098604
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French (fr)
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刘利新
梁慧怡
阎燕子
伍静娇
陈永明
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中山大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics

Definitions

  • the invention belongs to the field of macromolecules and biomedical materials, and relates to the use of cationic polymers or particles containing the polymers as effective ingredients to treat psoriasis.
  • Psoriasis is an immune-mediated chronic inflammatory skin disease. Its pathological features include abnormal proliferation of epidermal keratinocytes, dermal lymphocyte infiltration and vascular proliferation changes, and the affected area is often accompanied by erythema, scale and itching. symptom. At present, there are many treatments for psoriasis, but they all have certain drawbacks. Traditional treatment drugs such as commonly used glucocorticoid preparations can prevent the development of the disease and promote skin cell regeneration, but they are only suitable for skin lesions less than 10%.
  • cationic polymers and particles containing the polymers can reduce the activation of immune cells and reduce the concentration of cytokines, thereby alleviating the symptoms of psoriasis (see Figure 1), thereby providing the following invention:
  • the present invention provides the use of a polymer or particles containing the polymer as an active ingredient for the preparation of a medicine for the prevention or treatment of psoriasis, and the polymer is selected from the following cationic polymerization One or more of the following: poly-2-(dimethylamino)ethyl methacrylate (PDMA), poly- ⁇ -aminovalerolactone, polyetherimide (PEI), polyamide-amine dendritic Polymer (PAMAM-dendrimer), poly(2-dimethylaminoethylsulfide) caprolactone, polylysine (PLL), cationic starch, polycationic Q-glucan (ie Q-glucan ( Polycationic)), polyquaternary ammonium salt, polyacrylamide, protamine, Haidi membronium bromide (HDMBr), or the polymer is a cationic copolymer, which contains any one or more of the above cationic polymers The repeating unit.
  • the cationic copolymer is a cationic amphiphilic copolymer.
  • the cationic copolymer is a block copolymer, random copolymer, alternating copolymer, or graft copolymer.
  • the proportion of repeating units of any one or more of the above cationic polymers is not less than 30%, for example, 30%-50%, 50%-70%. % Or 70%-90%.
  • the cationic copolymer is a cationic block copolymer.
  • the cationic block copolymer comprises electrically neutral polymer segments and any one or more of the above cationic polymer segments.
  • the electrically neutral polymer segment is selected from one or more of the following polymer segments: polylactic acid-glycolic acid random copolymer (PLGA), hydrophobic polyphosphate, hydrophobic Polycarbonate, polyethylene glycol, polycaprolactone.
  • PLGA polylactic acid-glycolic acid random copolymer
  • hydrophobic polyphosphate hydrophobic Polyphosphate
  • hydrophobic Polycarbonate polyethylene glycol
  • polycaprolactone polycaprolactone
  • the cationic starch is 2-hydroxy-3-(trimethylamino)propyl ether starch chloride.
  • the polyquaternium salt is selected from:
  • a homopolymer of quaternary ammonium salt cationic monomers selected from the group consisting of dimethyl diallyl ammonium chloride, methacrylamide propyl trimethyl ammonium chloride and methacrylic acid Amidopropyl dodecyl dimethyl ammonium chloride;
  • the polyquaternium salt is selected from: polyquaternium-1, polyquaternium-2, polyquaternium-5, polyquaternium-6, polyquaternium-7 , Polyquaternium-10, Polyquaternium-11, Polyquaternium-12, Polyquaternium-13, Polyquaternium-14, Polyquaternium-16, Polyquaternium-17, Polyquaternium-19, Polyquaternium-20, Polyquaternium-22, Polyquaternium-27, Polyquaternium-28, Polyquaternium-29, Polyquaternium-30, Polyquaternium Quaternium-32, Polyquaternium-37, Polyquaternium-38, Polyquaternium-39, Polyquaternium-44, Polyquaternium-46, Polyquaternium-47, Polyquaternium Quaternium-51, Polyquaternium-53, Polyquaternium-55, Polyquaternium-58, Polyquaternium-60, Polyquaternium-67.
  • the cationic polymer or its segment has a number average molecular weight of 200-500, 500-5k, or 5k-500k, such as 200-300, 300-400, 400-500, 500-1k, 1k-5k, 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k- 200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • the cationic polymer or its segment has a weight average molecular weight of 200-500, 500-5k, or 5k-500k, such as 200-300, 300-400, 400-500, 500-1k, 1k-5k, 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k- 200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • the cationic polymer or its segment has a polymer dispersibility index (PDI) greater than 1 and less than 1.5, for example greater than 1 and less than 1.1, greater than 1 and less than 1.2, greater than 1 and less than 1.3 Or greater than 1 and less than 1.4.
  • PDI polymer dispersibility index
  • the electrically neutral polymer segment has a number average molecular weight of 5k-500k, such as 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k-200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • 5k-500k such as 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k-200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • the electrically neutral polymer segment has a weight average molecular weight of 5k-500k, such as 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k-200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • 5k-500k such as 5k-10k, 10k-20k, 20k-30k, 30k-40k, 40k-50k, 50k-60k, 60k-70k, 70k-80k, 80k-90k, 90k-100k, 100k-150k, 150k-200k, 200k-250k, 250k-300k, 300k-350k, 350k-400k, 400k-450k or 450k-500k.
  • the electrically neutral polymer segment has a polymer dispersibility index (PDI) greater than 1 and less than 1.5, for example greater than 1 and less than 1.1, greater than 1 and less than 1.2, greater than 1 and less than 1.3 Or greater than 1 and less than 1.4.
  • PDI polymer dispersibility index
  • the cationic block copolymer is a cationic amphiphilic block copolymer, which comprises a hydrophilic segment and a hydrophobic segment, wherein the hydrophilic segment is a cationic polymer segment,
  • the hydrophobic segment is an electrically neutral polymer segment.
  • the PLGA is an ester group (e.g., C 12 H 25 COO-) capped PLGA.
  • the polyethylene glycol is a methoxy-terminated polyethylene glycol.
  • the cationic polymer segments are PDMA segments.
  • the electrically neutral polymer segment is a PLGA segment.
  • the electrically neutral polymer segment is a C 12 H 25 COO-terminated PLGA segment.
  • the ratio of the number of lactic acid structural units to glycolic acid structural units is 90:10-10:90, such as 90:10-50:50 or 50:50-10: 90, such as 90:10, 75:25, 50:50, 25:70, or 10:90.
  • the number of lactic acid structural units is 10-100, such as 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90 or 90-100.
  • the number of glycolic acid structural units in the PLGA segment is 10-100, for example, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70 , 70-80, 80-90 or 90-100.
  • the degree of polymerization of the PDMA segment is 200-500, such as 200-250, 250-300, 300-350, 350-400, 400-450, or 450-500.
  • the hydrophilic cationic polymer segment is a PDMA segment
  • the hydrophobic, electrically neutral segment is a PLGA segment (for example, C 12 H 25 COO-terminated PLGA segment).
  • the number average molecular weight of the PDMA segment is 30k-80k (e.g., 30k-40k, 40k-50k, 50k-60k, 60k-70k, or 70k-80k).
  • the number average molecular weight of the PLGA segment is 8k-10k.
  • the numbers of lactic acid structural units and glycolic acid structural units are each independently selected from a positive integer of 80-100.
  • the particles comprising the polymer are nanoparticles or microparticles.
  • the particles have a particle size of 1 nm to 1 ⁇ m (e.g., 30 nm to 200 nm).
  • the particle size of the particles has a polydispersity index of 0.1-0.3, such as 0.1, 0.15, 0.2, 0.25, or 0.3.
  • the particles have a Zeta potential of +15mV to +20mV (e.g., +15mV, +16mV, +17mV, +18mV, +19mV, or +20mV).
  • the particles are spherical.
  • the particles are solid particles or micelles.
  • the particles are composed of the polymer.
  • the drug is prepared into any pharmaceutically acceptable dosage form, such as a transdermal formulation (such as ointment, plaster, patch, paint, aerosol, etc.) or intravenous injection Preparation (for example, liquid injection, powder for injection, or tablet for injection).
  • a transdermal formulation such as ointment, plaster, patch, paint, aerosol, etc.
  • intravenous injection Preparation for example, liquid injection, powder for injection, or tablet for injection.
  • the drug contains one or more pharmaceutically acceptable excipients (e.g., bases, excipients, carriers, stabilizers, or solubilizers).
  • pharmaceutically acceptable excipients e.g., bases, excipients, carriers, stabilizers, or solubilizers.
  • the pharmaceutically acceptable excipient is a carrier or excipient.
  • the carrier or excipient is selected from the group consisting of ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum proteins), sorbic acid, potassium sorbate, saturated plants Partial glyceride mixture of fatty acids, water, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose material, polyethylene glycol, carboxymethyl Base cellulose sodium, polyacrylate, beeswax, glycerin, gelatin, starch, polyethylene-polyoxypropylene block polymer and lanolin.
  • the adjuvant is a base, such as an ointment base, such as one or more of a grease base, a water-soluble base, and an emulsion base.
  • a base such as an ointment base, such as one or more of a grease base, a water-soluble base, and an emulsion base.
  • the ointment base is a water-soluble base, such as one or more selected from glycerin gelatin, starch glycerin, cellulose derivatives, and polyethylene glycols.
  • the glycerin gelatin is made by mixing and heating 10%-30% glycerin, 10%-15% gelatin and water.
  • the starch glycerin is made by mixing and heating 7%-10% starch, 70% glycerin and water.
  • the present invention provides a polymer micelle comprising a cationic amphiphilic copolymer (such as a cationic amphiphilic block copolymer) as defined in any one of the above, or It consists of a cationic amphiphilic copolymer (for example a cationic amphiphilic block copolymer) as defined above.
  • a cationic amphiphilic copolymer such as a cationic amphiphilic block copolymer
  • consists of a cationic amphiphilic copolymer for example a cationic amphiphilic block copolymer
  • the micelles have a particle size of 30nm-200nm, such as 30nm-50nm, 50nm-100nm, 100nm-150nm, or 150nm-200nm, such as 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm , 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm or 200nm.
  • 30nm-200nm such as 30nm-50nm, 50nm-100nm, 100nm-150nm, or 150nm-200nm, such as 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm , 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm or 200nm.
  • the particle size of the micelles has a polydispersity index of 0.1-0.3, such as 0.1, 0.15, 0.2, 0.25, or 0.3.
  • the micelles have a Zeta potential of +15mV to +20mV (e.g., +15mV, +16mV, +17mV, +18mV, +19mV, or +20mV).
  • the polymer micelles are spherical micelles.
  • the present invention provides a method for preparing the polymer micelle as described in any one of the above, including the following steps:
  • Step 1 Provide the cationic amphiphilic copolymer (for example, cationic amphiphilic block copolymer) as described in any one of the above;
  • Step 2 Self-assemble the cationic amphiphilic copolymer (for example, cationic amphiphilic block copolymer) in solution to form polymer micelles.
  • cationic amphiphilic copolymer for example, cationic amphiphilic block copolymer
  • the step 1 includes:
  • Step 1-1 Provide a hydrophobic electrically neutral polymer
  • Step 1-2 Making the hydrophobic electrically neutral polymer into a macromolecular initiator
  • Step 1-3 Use the macroinitiator obtained in step 1-2 to initiate polymerization of monomers to obtain a cationic amphiphilic block copolymer.
  • the step 1 also includes separation and/or purification operations (such as extraction, washing, precipitation or drying) of the hydrophobic electrically neutral polymer, macroinitiator or cationic amphiphilic block copolymer. ).
  • the step 2 includes:
  • Step 2-1 Dissolve the cationic amphiphilic copolymer in a solvent to obtain solution A;
  • Step 2-2 Drop solution A into the acidic solution, perform self-assembly, and obtain solution B;
  • Step 2-4 Adjust the pH of solution B after dialysis to obtain solution C;
  • Step 2-5 The solution C is concentrated by ultrafiltration.
  • the step 2 has one or more of the following characteristics:
  • step 2-1 The solvent in step 2-1 is tetrahydrofuran or dimethylformamide;
  • Step 2-2 is carried out under ultrasound
  • the acidic solution in step 2-2 is an aqueous hydrochloric acid solution
  • step 2-2 The pH of the acidic solution in step 2-2 is 2.5-4.5;
  • the concentration of solution B in step 2-2 is 1mg/mL-5mg/mL;
  • step 2-3 dialysis is performed for 24h-48h;
  • dialysis uses a dialysis bag with a molecular weight cut-off of 3000-4000 (for example, 3500);
  • step 2-3 dialysis is performed in dilute hydrochloric acid
  • step 2-3 dialysis is performed in dilute hydrochloric acid with a pH of 2.5-4.5;
  • steps 2-4 use potassium dihydrogen phosphate, disodium hydrogen phosphate and optionally sodium hydroxide to adjust the pH of the dialysis solution B;
  • steps 2-4 add potassium dihydrogen phosphate and disodium hydrogen phosphate until the total phosphate ion concentration is 0.01M-0.05M (for example, 0.01M);
  • step 2-5 ultrafiltration and concentration of solution C to a concentration of 15mg/mL-30mg/mL;
  • step 2-5 the centrifugation temperature of ultrafiltration and concentration is 0-5°C (for example, 4°C);
  • step 2-5 the centrifugal speed of ultrafiltration concentration is 2000 g/min-5000 g/min (for example, 4000 g/min).
  • the step 2 includes:
  • the centrifugal temperature of ultrafiltration concentration is 4°C
  • the centrifugal speed is 4000g/min.
  • the invention provides a composition comprising the polymer micelle of the invention.
  • the composition further includes a solvent, such as water.
  • the present invention provides the use of the above-mentioned polymer micelles in the preparation of a medicament for the prevention or treatment of psoriasis in a subject.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the polymer micelles of the present invention, and one or more pharmaceutically acceptable excipients (such as bases, excipients, carriers, stabilizers or enhancers). Solvent).
  • the pharmaceutically acceptable excipient is a carrier or excipient.
  • the carrier or excipient is selected from the group consisting of ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (for example, human serum proteins), sorbic acid, potassium sorbate, saturated plants Partial glyceride mixture of fatty acids, water, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose material, polyethylene glycol, carboxymethyl Base cellulose sodium, polyacrylate, beeswax, glycerin, gelatin, starch, polyethylene-polyoxypropylene block polymer and lanolin.
  • the adjuvant is a base, such as an ointment base, such as one or more of a grease base, a water-soluble base, and an emulsion base.
  • a base such as an ointment base, such as one or more of a grease base, a water-soluble base, and an emulsion base.
  • the ointment base is a water-soluble base, for example, one or more selected from glycerin gelatin, starch glycerin, cellulose derivatives, and polyethylene glycols.
  • the glycerin gelatin is made by mixing and heating 10%-30% glycerin, 10%-15% gelatin and water.
  • the starch glycerin is made by mixing and heating 7%-10% starch, 70% glycerin and water.
  • the pharmaceutical composition of the present invention can be made into any pharmaceutically acceptable dosage form.
  • the medicament or pharmaceutical composition of the present invention is formulated into a transdermal administration formulation (e.g., ointment, plaster, patch, paint, aerosol, etc.) or intravenous injection (e.g., liquid injection) , Powder for injection or tablet for injection).
  • a transdermal administration formulation e.g., ointment, plaster, patch, paint, aerosol, etc.
  • intravenous injection e.g., liquid injection
  • Powder for injection or tablet for injection e.g., Powder for injection or tablet for injection.
  • the present invention provides a method of preventing or treating psoriasis, comprising administering to a subject in need thereof a polymer as defined in any one of the above, particles comprising the polymer, polymer micelles Or pharmaceutical composition.
  • the method includes: administering the polymer, particle, polymer micelle or pharmaceutical composition to the subject by transdermal administration (e.g., 50 mg/kg-250 mg /kg to the subject).
  • transdermal administration e.g., 50 mg/kg-250 mg /kg to the subject.
  • the method includes: administering the polymer, particle, polymer micelle or pharmaceutical composition to the subject by intravenous injection (e.g., 1 mg/kg-6.25 mg/ The dose of kg was administered to the subject).
  • intravenous injection e.g., 1 mg/kg-6.25 mg/ The dose of kg was administered to the subject.
  • the method includes: administering the polymer, particle, polymer micelle or pharmaceutical composition once a day for 5-7 days.
  • the present invention provides a polymer, particle, polymer micelle or pharmaceutical composition as defined in any one of the above, which is used to prevent or treat psoriasis.
  • the polymer, particle, polymer micelle or pharmaceutical composition is administered to the subject by transdermal administration (for example, administered at a dose of 50 mg/kg-250 mg/kg). To subjects).
  • the polymer, particle, polymer micelle or pharmaceutical composition is administered to the subject by intravenous injection (for example, administered at a dose of 1 mg/kg-6.25 mg/kg Subject).
  • the polymer, particle, polymer micelle or pharmaceutical composition is administered once a day for 5-7 days.
  • the polymer, particle or polymer micelle can be administered to a subject as a single active ingredient, or can be co-administered with other active ingredients for the treatment of psoriasis.
  • the subject is preferably a mammal, such as bovine, equine, swine, canine, feline, rodent, primate, for example, human.
  • block copolymer refers to a linear polymer composed of two or more structurally different segments.
  • cationic amphiphilic block copolymer refers to a copolymer that is (mainly) composed of a hydrophilic segment and a hydrophobic segment that has a positive charge or carries a positive charge after being ionized in water.
  • particle refers to a state of matter characterized by the presence of discrete particles, pellets, beads, or pellets, regardless of their size, shape, or morphology.
  • nanoparticle and “microparticle” refer to particles with a particle size of nanometer or micrometer, for example, particles with a particle size of 1nm-1000nm or 1 ⁇ m-1000 ⁇ m, which can be solid particles or Particles in the form of micelles or vesicles.
  • polymer micelle refers to an amphiphilic polymer (e.g., amphiphilic block copolymer) in a solvent (e.g., water), when its concentration exceeds a certain critical value, due to
  • the solvophobic part (for example, the hydrophobic part) or the solvophilic part (for example, the hydrophilic part) attract each other to form a solid core-shell structure whose core is composed of the solvophobic part of the molecule (for example, the hydrophobic part)
  • the shell is composed of the solvophilic part (for example, the hydrophilic part) of the molecule.
  • a solution self-assembly method can be used to form polymer micelles.
  • particle size refers to “equivalent particle size”, which refers to when a certain physical property or physical behavior of the measured particle is closest to a homogeneous sphere (or combination) of a certain diameter, Just take the diameter (or combination) of the sphere as the equivalent particle size (or particle size distribution) of the measured particle.
  • pharmaceutically acceptable excipients refers to those used in the production of drugs and formulating prescriptions, except for the active ingredients, which have been reasonably evaluated in terms of safety, and are included in pharmaceutical preparations. Substances, which can have one or more of the functions of shaping, acting as a carrier, improving stability, solubilizing, solubilizing, and slow and controlled release.
  • polyquaternary ammonium salt refers to a polymer with a quaternary ammonium salt structure on all or part of the repeating unit, which usually includes semi-synthetic polyquaternary ammonium salts (such as natural polymers (such as cellulose) that have undergone quaternization modification The obtained polymer), and synthetic polyquaternary ammonium salt (for example, a polymer synthesized by homopolymerization of cationic monomers containing double bonds or copolymerization with other monomers).
  • semi-synthetic polyquaternary ammonium salts such as natural polymers (such as cellulose) that have undergone quaternization modification The obtained polymer
  • synthetic polyquaternary ammonium salt for example, a polymer synthesized by homopolymerization of cationic monomers containing double bonds or copolymerization with other monomers.
  • the present invention has discovered that cationic polymers can reduce the activation of immune cells, reduce the concentration of cytokines, and have obvious therapeutic effects on psoriasis.
  • the invention also prepares micelles containing cationic amphiphilic polymers, which have obvious therapeutic effects on psoriasis.
  • Figure 1 exemplarily describes the mechanism and process of cationic polymer or its particles in the treatment of psoriasis.
  • cationic polymer can enter the affected skin by transdermal administration methods including but not limited to drip coating, reducing
  • the activation of immune cells reduces the concentration of cytokines, relieves the symptoms of psoriasis, and plays a therapeutic role.
  • Figure 2 shows the structural formula (a), 1 HNMR spectrum (b) and GPC elution time curve (c) of the amphiphilic block copolymer PLGA-b-PDMA 474 (all with PDMA 480 as a reference).
  • Figure 3 shows the particle size distribution diagram (a) and TEM photograph (b) of the cationic polymer micelle cNP.
  • Figure 4 shows the PASI score of the back skin of mice in each group in the in vivo efficacy experiment of transdermal administration.
  • Figure 5 shows the H&E staining results of pathological sections of the back skin of mice in each group in the in vivo efficacy experiment of transdermal administration.
  • Figure 6 shows the results of the PASI score of the back skin of each group of mice in the in vivo efficacy experiment of intravenous injection.
  • Figure 7 shows the H&E staining results of pathological sections of the back skin of mice in each group in the in vivo efficacy experiment of intravenous injection.
  • Figure 8 shows the contents of various immune factors in the back skin of mice in each group after treatment in the in vivo efficacy experiment.
  • Figure 9 shows the results of the PASI score of the back skin of the psoriasis cynomolgus monkey in the in vivo efficacy experiment of transdermal administration of cNP.
  • Figure 10 shows photos of the skin of cynomolgus monkeys with psoriasis on day 0, and from day 8 to day 13.
  • Figure 11 shows the H&E staining results of the pathological section of the back skin of the psoriasis cynomolgus monkey on the 13th day after treatment for five days in the in vivo efficacy experiment of cNP percutaneous administration.
  • Figure 12 shows the results of liver function and kidney function tests of mice in each group.
  • Figure 13 shows the H&E staining results of pathological sections of the heart, liver, spleen, lung, and kidney of each group of mice.
  • Figure 14 shows the changes in liver and kidney functions of cynomolgus monkeys before and 7 days after applying cNP paint.
  • the cationic amphiphilic block copolymer used to prepare micelles in this example is PLGA-b-PDMA 474 , and its structural formula is as follows:
  • the product obtained by S2 was extracted once with 1mol/L dilute hydrochloric acid, twice with 1mol/L sodium bicarbonate, and once with saturated sodium chloride solution. After the organic phase was dried with anhydrous MgSO 4 , it was rotated After steaming to 20mL, it was precipitated three times in a mixed precipitation agent of 200mL cold ether and cold methanol, and the precipitate was dried in a vacuum oven to obtain the macromolecular initiator PLGA-Br;
  • Figure 2 shows the proton nuclear magnetic resonance ( 1 HNMR) spectrum of PLGA-b-PDMA 474 and the gel permeation chromatography (GPC) elution time curve (both are based on PDMA 470 , and the preparation process of PDMA 470 is shown in Example 2).
  • 1 HNMR proton nuclear magnetic resonance
  • GPC gel permeation chromatography
  • cNP cationic polymer micelles
  • the prepared cationic polymer micelle cNP has an average particle size of 44nm and a Zeta potential of +18mV.
  • Figure 3a is a particle size distribution diagram of cNP measured by dynamic light scattering (DLS), and Figure 3b is a transmission electron microscope (TEM) photo of cNP.
  • cNPs are spherical micelles with relatively uniform particle size distribution.
  • the PBS solution containing cNP was concentrated by ultrafiltration and centrifugation to 50 mg/mL, and then mixed with a PBS solution containing 30% gelatin and 20% glycerol at a volume ratio of 1:1 to obtain 25 mg/mL cNP, 15% gelatin A cNP paint with a certain viscosity with 10% glycerin.
  • the water-soluble cationic polymer PDMA 480 is prepared by referring to the method of Macromolecules 31, 5167-5169 (1998), and its structural formula is as follows:
  • Figure 2 shows the proton nuclear magnetic resonance ( 1 HNMR) spectrum and the gel permeation chromatography (GPC) elution time curve.
  • Example 1 a PDMA-based paint was prepared.
  • PAMAM Generation 3 dendrimer (PAMAM-G3), molecular weight 6900, purchased from Sigma and post-processed (product name PAMAM dendrimer, ethylenediamine core, generation 3.0 solution 20wt.% in methanol), the polymer solution after purchase After dialysis in pure water with a dialysis bag with a molecular weight cut-off of 3500 for 24 hours, it was freeze-dried to obtain PAMAM-G3 solid.
  • a coating agent based on PAMAM-G3 was prepared.
  • Protamine purchased from Aladdin. Referring to the preparation method of Example 1, a protamine-based paint was prepared.
  • Polyquaternium-10 purchased from Shanghai Anyan Trading Co., Ltd. Referring to the preparation method of Example 1, a coating agent based on polyquaternium-10 was prepared.
  • Polylysine (PLL) is prepared by referring to the method of J. Am. Chem. Soc. 1999, 121, 5919-5929, and the structure is as follows:
  • Example 1 a polylysine-based paint was prepared.
  • Grouping Divide Bal B/C mice weighing 20g into one model group, one blank group, and two intravenous injection groups (ie, PDMA(iv) administration group and cNP(iv) administration Group), eight transdermal administration groups (ie, PDMA paint administration group, cNP paint administration group, protamine paint administration group, polyquaternium salt administration group, Haiti membronium bromide The paint administration group, the PAMAM-G3 paint administration group and the polylysine paint administration group), each with 10 animals.
  • intravenous injection groups ie, PDMA(iv) administration group and cNP(iv) administration Group
  • transdermal administration groups ie, PDMA paint administration group, cNP paint administration group, protamine paint administration group, polyquaternium salt administration group, Haiti membronium bromide
  • the paint administration group, the PAMAM-G3 paint administration group and the polylysine paint administration group each with 10 animals.
  • mice are anesthetized with isoflurane gas, the hair on the back 2cm*2cm skin is shaved with a razor on the first day, and 62.5mg of imiquine of Idalla is applied to this skin for seven consecutive days Mott cream was absorbed to induce psoriasis-like skin lesions in mice. The psoriasis model was established seven days after application.
  • Transdermal administration On the 8th day when the model is formed, that is, when the psoriasis mice have redness and/or scaly symptoms, apply 250mg/kg or 1.25mg/cm 2 to the corresponding group of different paints. Put gauze on the skin of the rat's back and fix it with a bandage. The mice were administered percutaneously for 5 consecutive days; the mice in the model group were smeared with a glycerin gelatin base on the back skin as a control.
  • Intravenous administration On the 8th day when the model is established, that is, when the psoriasis mice have redness, swelling and/or scaly symptoms, pass the cNP micellar PBS solution or PDMA 480 PBS solution through the tail vein at a dose of 12.5 mg/kg, respectively It was injected into mice; mice were administered via tail vein injection for 5 consecutive days; mice in the model group were injected with an equal volume of PBS via tail vein as a control.
  • the PCR method was used for semi-quantitative analysis of the mRNA levels of TNF- ⁇ , IL-6, IL-17A, IL17-R, IL22, and IL23.
  • Figure 4 shows the PASI score of the back skin of mice in each group in the in vivo efficacy experiment of transdermal administration. among them:
  • Figure 4a shows the blank group, the model group, the PDMA paint administration group, the cNP paint administration group, the protamine paint administration group, the polyquaternium salt administration group, the Haitian membronium bromide paint administration group
  • the drug group PAMAM-G3 paint administration group, polylysine paint administration group, scores for the degree of redness and swelling of the back skin from day 1 to day 13;
  • Figure 4b shows the blank group, the model group, the PDMA paint administration group, the cNP paint administration group, the protamine paint administration group, the polyquaternium salt administration group, and the Haitian membronium bromide paint administration group.
  • Figure 4c shows the blank group, the model group, the PDMA paint administration group, the cNP paint administration group, the protamine paint administration group, the polyquaternium salt administration group, the Haitian membronium bromide paint administration group
  • Figure 4d shows the blank group, the model group, the PDMA paint administration group, the cNP paint administration group, the protamine paint administration group, the polyquaternium salt administration group, the Haitian membronium bromide paint administration group
  • the total scores of the back skin of the drug group, PAMAM-G3 paint administration group, and polylysine paint administration group from day 1 to day 13.
  • Figure 5 shows the H&E staining results of pathological sections of the back skin of mice in each group in the in vivo efficacy experiment of transdermal administration, including the blank group, model group, PDMA paint administration group, and cNP paint administration group. H&E staining of the protamine paint administration group, the polyquaternium salt paint administration group, the Haitian mebromide paint administration group, the PAMAM-G3 paint administration group, and the polylysine paint administration group result.
  • Figure 6 shows the PASI score results of the back skin of each group of mice in the in vivo efficacy experiment of intravenous injection, where:
  • Figure 6a shows the blank group, the model group, the PDMA (i.v.) administration group, and the cNP (i.v.) administration group was scored by the degree of skin redness and swelling from day 1 to day 13;
  • Figure 6b shows the back skin scale scores of the blank group, model group, PDMA (i.v.) administration group, and cNP (i.v.) administration group from day 1 to day 13;
  • Figure 6c shows the back skin thickness scores of the blank group, model group, PDMA (i.v.) administration group, and cNP (i.v.) administration group from day 1 to day 13;
  • Figure 6d shows the total score of the back skin of the blank group, model group, PDMA (i.v.) administration group, and cNP (i.v.) administration group from day 1 to day 13.
  • Figure 7 shows the H&E staining results of pathological sections of the back skin of mice in each group in the in vivo efficacy experiment of intravenous injection, including blank group, model group, PDMA(iv) administration group, and cNP(iv) administration The staining result of the group.
  • Figure 8 shows the in vivo efficacy experiment, the blank group, model group, PDMA paint administration group, cNP paint administration group, PDMA(iv) administration group, and cNP(iv) administration group on the 13th day.
  • PDMA paint administration group After five days of treatment, PDMA paint administration group, cNP paint administration group, PDMA(iv) administration group, cNP(iv) administration group TNF- ⁇ , IL-6, IL-17A, IL-17F,
  • PDMA(iv) administration group After five days of treatment, PDMA(iv) administration group, cNP(iv) administration group TNF- ⁇ , IL-6, IL-17A, IL-17F,
  • TNF- ⁇ IL-6
  • IL-17A IL-17F
  • experiment (2) intends to use primate cynomolgus monkeys to conduct experiments to better evaluate the therapeutic effects of cationic polymers or micelles.
  • Cynomolgus monkeys are similar to humans in their metabolism, function and structure, so they are a better model animal for studying human-related diseases.
  • Using cynomolgus monkeys as experimental animals to conduct animal experiments can more realistically and effectively evaluate the actual efficacy of cationic polymers or their micelles, and evaluate the feasibility of treatment of actual human diseases.
  • Grouping 3 cynomolgus monkeys, male, weighing about 4-5kg, and the feeding period is 8 weeks.
  • Modeling After shaving each cynomolgus monkey, apply about 1g of imiquimod cream to the skin of 2cm*2cm on the upper and lower sides of the animal's back to make a model. Apply until the skin is basically absorbed, and apply for 7 days. Later the model is established.
  • Figure 9 shows the results of the PASI score of the back skin of the psoriasis cynomolgus monkey in the in vivo efficacy experiment of transdermal administration of cNP. among them:
  • Figure 9a shows the back skin redness and swelling scores of the blank group, model group, and cNP paint administration group from day 1 to day 13;
  • Figure 9b shows the back skin scale scores of the blank group, model group, and cNP paint administration group from day 1 to day 13;
  • Figure 9c shows the back skin thickness scores of the blank group, model group, and cNP paint administration group from day 1 to day 13;
  • Figure 9d shows the total score of the back skin of the blank group, model group, and cNP paint administration group from day 1 to day 13.
  • Figure 10 shows photos of the skin of cynomolgus monkeys with psoriasis on day 0, day 8 to day 13.
  • Three of the cynomolgus monkeys (No. 0126, 0148 and 0150) took photos of the cNP-treated psoriasis skin on the lower back of the back and the self-control model skin on the upper side on day 0, day 8 to day 13.
  • the photo on day 0 is the photo of the skin before the model is created
  • the photo on the 9th to the 13th day is the photo of the skin after administration.
  • the yellow dashed box is the range of 2cm*2cm selected during model building.
  • Figure 11 shows the H&E staining results of the pathological section of the back skin of the psoriasis cynomolgus monkey on the 13th day after treatment for five days in the in vivo efficacy experiment of transdermal administration of cNP, including the blank group (normal monkey skin), H&E staining results of model group (skin not coated with cNP on the upper side of psoriasis monkeys) and cNP paint administration group (skin coated with cNP on the underside of psoriasis monkeys).
  • the big picture is a picture magnified 4 times, and the small picture is a picture magnified 20 times.
  • the experimental process is shown in the process of experiment (1) in experimental example 1.
  • the blank group model group, PDMA paint administration group, cNP paint administration group, PDMA(iv) administration group, cNP(iv) The drug group received blood from the orbit, centrifuged the blood to take the upper serum, and measured the indicators of alkaline phosphatase (ALP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) reflecting liver function with an automatic biochemical analyzer and reflecting renal function Of uric acid, urea, and creatinine.
  • ALP alkaline phosphatase
  • ALT alanine aminotransferase
  • AST aspartate aminotransferase
  • mice in each group were euthanized on the 13th day, and the heart, liver, spleen, lung, and viscera were taken for paraffin sections and H&E stained.
  • Figure 12 shows the results of liver function and kidney function tests of mice in each group.
  • the liver and kidney functions of the cNP administration group are closer to those of the normal group, while the PDMA group has a certain gap from the normal group of mice.
  • Such as PDMA paint group AST, ALT, uric acid, urea are all higher.
  • the results show that cNP is less hepatotoxic and nephrotoxic than PDMA.
  • Figure 13 shows the H&E staining results of pathological sections of the heart, liver, spleen, lung, and kidney of each group of mice.
  • the cNP administration group had no obvious damage to internal organs regardless of intravenous or transdermal administration, while the PDMA injection group caused hepatocyte apoptosis, alveolar hemorrhage, glomerular atrophy and other symptoms
  • the PDMA transdermal administration group caused symptoms such as hepatocellular edema and glomerular atrophy.
  • the results showed that cNP is less toxic to internal organs than PDMA.
  • Figure 14 shows the changes in liver and kidney functions of cynomolgus monkeys before and 7 days after applying cNP paint. As shown in the figure, the cynomolgus monkeys did not have much changes in ALP, AST, ALT, creatinine, uric acid, and urea after applying cNP paint 7 days after applying cNP paint. Obvious in vivo toxicity, and will not affect normal liver and kidney functions.

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Abstract

聚合物或包含所述聚合物的颗粒作为有效成分用于制备预防或治疗银屑病的药物的用途，聚合物胶束及其制备方法和药物组合物。所述聚合物为选自聚甲基丙烯酸2-(二甲氨基)乙酯、聚α-氨基戊内酯、聚醚酰亚胺、聚酰胺-胺型树枝状聚合物、聚(2-二甲氨基乙基硫)己内酯、聚赖氨酸、阳离子淀粉、聚阳离子型Q-葡聚糖、聚季铵盐、聚丙烯酰胺、鱼精蛋白、海地美溴铵的阳离子聚合物，或包含以上任意一种或多种阳离子聚合物重复单元的阳离子型共聚物。

Description

可用于治疗银屑病的阳离子聚合物及颗粒 技术领域
本发明属于高分子和生物医学材料领域，涉及阳离子聚合物或包含所述聚合物的颗粒作为有效成分治疗银屑病的用途。
背景技术
银屑病(psoriasis)是一种免疫介导的慢性炎症性皮肤病，其病理特征包括表皮角质形成细胞异常增殖，真皮淋巴细胞浸润和血管增生性改变，患处多伴有红斑、鳞屑及瘙痒等症状。目前治疗银屑病的疗法有很多，但都存在一定的弊端，传统治疗药物例如常用的糖皮质激素制剂，虽然可阻止病情发展并促进皮肤细胞再生色素，但只适用于皮损面积<10％的患者，且激素类药物长期使用容易产生依赖性，停药后常迅速复发甚至加重，并且还会产生应激性溃疡、高血压、糖尿病、骨质疏松等副作用；而一些生物制剂如抗IL-12和IL-23药物优特克单抗、以及抗IL-17的单克隆抗体药物苏金单抗则存在价格昂贵等缺点，限制了这类药物的广泛使用。因此，亟需开发更加有效、低毒、廉价的抗银屑病药物来满足市场需求，为银屑病的治疗提供新策略。
发明内容
本发明的发明人发现，阳离子聚合物及包含所述聚合物的颗粒可减少免疫细胞的激活，降低细胞因子的浓度，从而缓解银屑病的症状(参见图1)，由此提供了下述发明：
在一个方面，本发明提供了以聚合物或包含所述聚合物的颗粒作为有效成分用于制备药物的用途，所述药物用于预防或治疗银屑病，所述聚合物选自以下阳离子聚合物中的一种或多种：聚甲基丙烯酸2-(二甲氨基)乙酯(PDMA)、聚α-氨基戊内酯、聚醚酰亚胺(PEI)、聚酰胺-胺型树枝状聚合物(PAMAM-dendrimer)、聚(2-二甲氨基乙基硫)己内酯、聚赖氨酸(PLL)、阳离子淀粉、聚阳离子型Q-葡聚糖(即Q-葡聚糖(聚阳离子型))、聚季铵盐、聚丙烯酰胺、鱼精蛋白、海地美溴铵(HDMBr)，或者所述聚合物为阳离子型共聚物，其包含以上任意一种或多种阳离子聚合物的重复单元。
在某些实施方案中，所述阳离子型共聚物为阳离子型两亲性共聚物。
在某些实施方案中，所述阳离子型共聚物为嵌段共聚物、无规共聚物、交替共聚物 或接枝共聚物。
在某些实施方案中，所述阳离子型共聚物中，以上任意一种或多种阳离子聚合物的重复单元所占的比例不低于30％，例如为30％-50％、50％-70％或70％-90％。
在某些实施方案中，所述阳离子型共聚物为阳离子型嵌段共聚物。
在某些实施方案中，所述阳离子型嵌段共聚物包含电中性聚合物链段和以上任意一种或多种阳离子聚合物的链段。
在某些实施方案中，所述电中性聚合物链段选自以下一种或多种聚合物的链段：聚乳酸-羟基乙酸无规共聚物(PLGA)、疏水性聚磷酸酯、疏水性聚碳酸酯、聚乙二醇、聚己内酯。
在某些实施方案中，所述阳离子淀粉为2-羟基-3-(三甲基氨根)丙基醚淀粉氯化物。
在某些实施方案中，所述聚季铵盐选自：
阳离子纤维素；
阳离子瓜尔胶；
季铵盐类阳离子单体的均聚物，所述季铵盐类阳离子单体选自二甲基二烯丙基氯化铵、甲基丙烯酰胺丙基三甲基氯化铵和甲基丙烯酰胺丙基十二烷基二甲基氯化铵；
所述季铵盐类阳离子单体之间形成的共聚物；以及
一种或多种所述季铵盐类阳离子单体与一种或多种其他单体形成的共聚物，所述其他单体选自：丙烯酰胺、丙烯酸、丙烯酸酯、乙烯基吡咯烷酮及其任意组合。
在某些实施方案中，所述聚季铵盐选自：聚季铵盐-1、聚季铵盐-2、聚季铵盐-5、聚季铵盐-6、聚季铵盐-7、聚季铵盐-10、聚季铵盐-11、聚季铵盐-12、聚季铵盐-13、聚季铵盐-14、聚季铵盐-16、聚季铵盐-17、聚季铵盐-19、聚季铵盐-20、聚季铵盐-22、聚季铵盐-27、聚季铵盐-28、聚季铵盐-29、聚季铵盐-30、聚季铵盐-32、聚季铵盐-37、聚季铵盐-38、聚季铵盐-39、聚季铵盐-44、聚季铵盐-46、聚季铵盐-47、聚聚季铵盐-51、聚季铵盐-53、聚季铵盐-55、聚季铵盐-58、聚季铵盐-60、聚季铵盐-67。
在某些实施方案中，所述阳离子聚合物或其链段具有200-500、500-5k或5k-500k的数均分子量，例如200-300、300-400、400-500、500-1k、1k-5k、5k-10k、10k-20k、20k-30k、30k-40k、40k-50k、50k-60k、60k-70k、70k-80k、80k-90k、90k-100k、100k-150k、150k-200k、200k-250k、250k-300k、300k-350k、350k-400k、400k-450k或450k-500k。
在某些实施方案中，所述阳离子聚合物或其链段具有200-500、500-5k或5k-500k 的重均分子量，例如200-300、300-400、400-500、500-1k、1k-5k、5k-10k、10k-20k、20k-30k、30k-40k、40k-50k、50k-60k、60k-70k、70k-80k、80k-90k、90k-100k、100k-150k、150k-200k、200k-250k、250k-300k、300k-350k、350k-400k、400k-450k或450k-500k。
在某些实施方案中，所述阳离子聚合物或其链段具有大于1且小于1.5的聚合物分散性指数(PDI)，例如大于1且小于1.1、大于1且小于1.2、大于1且小于1.3或大于1且小于1.4。
在某些实施方案中，所述电中性聚合物链段具有5k-500k的数均分子量，例如5k-10k、10k-20k、20k-30k、30k-40k、40k-50k、50k-60k、60k-70k、70k-80k、80k-90k、90k-100k、100k-150k、150k-200k、200k-250k、250k-300k、300k-350k、350k-400k、400k-450k或450k-500k。
在某些实施方案中，所述电中性聚合物链段具有5k-500k的重均分子量，例如5k-10k、10k-20k、20k-30k、30k-40k、40k-50k、50k-60k、60k-70k、70k-80k、80k-90k、90k-100k、100k-150k、150k-200k、200k-250k、250k-300k、300k-350k、350k-400k、400k-450k或450k-500k。
在某些实施方案中，所述电中性聚合物链段具有大于1且小于1.5的聚合物分散性指数(PDI)，例如大于1且小于1.1、大于1且小于1.2、大于1且小于1.3或大于1且小于1.4。
在某些实施方案中，所述阳离子型嵌段共聚物为阳离子型两亲性嵌段共聚物，其包含亲水段和疏水段，其中，所述亲水段为阳离子聚合物的链段，所述疏水段为电中性聚合物链段。
在某些实施方案中，所述PLGA为酯基(例如C 12H 25COO-)封端的PLGA。
在某些实施方案中，所述聚乙二醇为甲氧基封端的聚乙二醇。
在某些实施方案中，所述阳离子聚合物的链段为PDMA链段。
在某些实施方案中，所述电中性聚合物链段为PLGA链段。
在某些实施方案中，所述电中性聚合物链段为C 12H 25COO-封端的PLGA链段。
在某些实施方案中，所述PLGA链段中，乳酸结构单元和羟基乙酸结构单元的个数比为90:10-10:90，例如90:10-50:50或50:50-10:90，例如90:10、75:25、50:50、25:70或10:90。
在某些实施方案中，所述PLGA链段中，乳酸结构单元的个数为10-100，例如10-20、 20-30、30-40、40-50、50-60、60-70、70-80、80-90或90-100。
在某些实施方案中，所述PLGA链段中，羟基乙酸结构单元的个数为10-100，例如10-20、20-30、30-40、40-50、50-60、60-70、70-80、80-90或90-100。
在某些实施方案中，所述PDMA链段的聚合度为200-500，例如200-250、250-300、300-350、350-400、400-450或450-500。
在某些实施方案中，所述阳离子型两亲性嵌段共聚物中，亲水的阳离子聚合物链段为PDMA链段，疏水的电中性链段为PLGA链段(例如C 12H 25COO-封端的PLGA链段)。在某些实施方案中，PDMA链段的数均分子量为30k-80k(例如30k-40k、40k-50k、50k-60k、60k-70k或70k-80k)。在某些实施方案中，PLGA链段的数均分子量为8k-10k。在某些实施方案中，PLGA链段中，乳酸结构单元和羟基乙酸结构单元的个数各自独立地选自80-100的正整数。
在某些实施方案中，包含所述聚合物的颗粒为纳米颗粒或微米颗粒。
在某些实施方案中，所述颗粒具有1nm-1μm(例如30nm-200nm)的粒径。
在某些实施方案中，所述颗粒的粒径具有0.1-0.3的多分散指数，例如0.1、0.15、0.2、0.25或0.3。
在某些实施方案中，所述颗粒具有+15mV至+20mV(例如+15mV、+16mV、+17mV、+18mV、+19mV或+20mV)的Zeta电位。
在某些实施方案中，所述颗粒为球形。
在某些实施方案中，所述颗粒为实心颗粒或胶束。
在某些实施方案中，所述颗粒由所述聚合物组成。
在某些实施方案中，所述药物被制成药学上可接受的任一剂型，例如经皮给药制剂(例如软膏剂、硬膏剂、贴剂、涂剂、气雾剂等)或静脉注射制剂(例如液体注射剂、注射用粉剂或注射用片剂)。
在某些实施方案中，所述药物包含一种或多种药学上可接受的辅料(例如基质、赋形剂、载体、稳定剂或增溶剂)。
在某些实施方案中，所述药学上可接受的辅料为载体或赋形剂。
在某些实施方案中，所述载体或赋形剂选自：离子交换剂，氧化铝，硬脂酸铝，卵磷脂，血清蛋白(例如人血清蛋白)，山梨酸，山梨酸钾，饱和植物脂肪酸的部分甘油酯混合物，水，磷酸氢二钠，磷酸氢钾，氯化钠，锌盐，胶态氧化硅，三硅酸镁，聚乙 烯吡咯烷酮，纤维素物质，聚乙二醇，羧甲基纤维素钠，聚丙烯酸酯，蜂蜡，甘油，明胶，淀粉，聚乙烯-聚氧丙烯嵌段聚合物和羊毛脂。
在某些实施方案中，所述辅料为基质，例如软膏剂基质，例如油脂性基质、水溶性基质和乳剂型基质的一种或多种。
在某些实施方案中，所述软膏剂基质为水溶性基质，例如选自甘油明胶、淀粉甘油、纤维素衍生物类、聚乙二醇类的一种或多种。在某些实施方案中，所述甘油明胶由10％-30％甘油、10％-15％明胶以及水经混合加热制成。在某些实施方案中，所述淀粉甘油由7％-10％的淀粉、70％甘油与水经混合加热制成。
在一个方面，本发明提供了一种聚合物胶束，所述聚合物胶束包含如上任一项定义的阳离子型两亲性共聚物(例如阳离子型两亲性嵌段共聚物)，或由如上定义的阳离子型两亲性共聚物(例如阳离子型两亲性嵌段共聚物)组成。
在某些实施方案中，所述胶束具有30nm-200nm的粒径，例如30nm-50nm、50nm-100nm、100nm-150nm或150nm-200nm，例如30nm、40nm、50nm、60nm、70nm、80nm、90nm、100nm、110nm、120nm、130nm、140nm、150nm、160nm、170nm、180nm、190nm或200nm。
在某些实施方案中，所述胶束的粒径具有0.1-0.3的多分散指数，例如0.1、0.15、0.2、0.25或0.3。
在某些实施方案中，所述胶束具有+15mV至+20mV(例如+15mV、+16mV、+17mV、+18mV、+19mV或+20mV)的Zeta电位。
在某些实施方案中，所述聚合物胶束为球形胶束。
在一个方面，本发明提供了制备如上任一项所述的聚合物胶束的方法，包括以下步骤：
步骤1：提供如上任一项所述的阳离子型两亲性共聚物(例如阳离子型两亲性嵌段共聚物)；
步骤2：将所述阳离子型两亲性共聚物(例如阳离子型两亲性嵌段共聚物)在溶液中进行自组装，形成聚合物胶束。
在某些实施方案中，所述步骤1包括：
步骤1-1：提供疏水的电中性聚合物；
步骤1-2：将所述疏水的电中性聚合物制成大分子引发剂；
步骤1-3：以步骤1-2得到的大分子引发剂引发单体进行聚合，得到阳离子型两亲性嵌段共聚物。
任选地，所述步骤1还包括对疏水的电中性聚合物、大分子引发剂或阳离子型两亲性嵌段共聚物进行分离和/或纯化的操作(例如萃取、洗涤、沉淀或干燥)。
在某些实施方案中，所述步骤2包括：
步骤2-1：将阳离子型两亲性共聚物溶于溶剂中，得到溶液A；
步骤2-2：将溶液A滴入酸性溶液中，进行自组装，得到溶液B；
步骤:2-3：对溶液B进行透析；
步骤2-4：对透析后的溶液B的pH进行调节，得到溶液C；
步骤2-5：对溶液C进行超滤浓缩。
在某些实施方案中，所述步骤2具有以下特征中的一个或多个：
(1)步骤2-1中的溶剂为四氢呋喃或二甲基甲酰胺；
(2)步骤2-1中的溶液A的浓度为5mg/mL-15mg/mL；
(3)步骤2-2在超声作用下进行；
(4)步骤2-2中的酸性溶液为盐酸水溶液；
(5)步骤2-2中的酸性溶液的pH值为2.5-4.5；
(6)步骤2-2中溶液B的浓度为1mg/mL-5mg/mL；
(7)步骤2-3中，透析进行24h-48h；
(8)步骤2-3中，透析使用截留分子量为3000-4000(例如3500)的透析袋；
(9)步骤2-3中，透析在稀盐酸中进行；
(10)步骤2-3中，透析在pH值为2.5-4.5的稀盐酸中进行；
(11)步骤2-4中，使用磷酸二氢钾、磷酸氢二钠和任选的氢氧化钠对透析后的溶液B的pH进行调节；
(12)步骤2-4中，加入磷酸二氢钾和磷酸氢二钠至磷酸根离子总浓度为0.01M-0.05M(例如0.01M)；
(13)步骤2-4中，溶液C的pH为7-8(例如7.4)；
(14)步骤2-5中，将溶液C超滤浓缩至浓度为15mg/mL-30mg/mL；
(15)步骤2-5中，超滤浓缩的离心温度为0-5℃(例如4℃)；和
(16)步骤2-5中，超滤浓缩的离心速度为2000g/min-5000g/min(例如4000g/min)。
在某些实施方案中，所述步骤2包括：
S1.将阳离子型两亲性共聚物加入四氢呋喃中，溶解，配成5mg/mL-15mg/mL的溶液A；
S2.在超声破碎仪的超声作用下，将溶液A逐滴缓慢加入到稀盐酸水溶液中，自组装得到1mg/mL-5mg/mL的溶液B；
S3.将溶液B装入截留分子量为3500的透析袋，在稀盐酸中透析24h-48h；
S4.收集透析后的溶液B，往里面加入磷酸二氢钾、磷酸氢二钠、氢氧化钠，调节pH至7.4得C溶液；
S5.使用超滤离心管浓缩溶液C，浓缩到15-30mg/mL；
其中，S2、S3中，稀盐酸的pH值为2.5-4.5；
S4中，加入磷酸二氢钾和磷酸氢二钠至磷酸根离子总浓度为0.01M；
S5中，超滤浓缩的离心温度为4℃，离心速度为4000g/min。
在一个方面，本发明提供了一种组合物，其包含本发明的聚合物胶束。
在某些实施方案中，所述组合物还包含溶剂，例如水。
在一个方面，本发明提供了上述聚合物胶束在制备药物中的用途，所述药物用于预防或治疗受试者的银屑病。
在一个方面，本发明提供了一种药物组合物，其包含本发明的聚合物胶束，以及一种或多种药学上可接受的辅料(例如基质、赋形剂、载体、稳定剂或增溶剂)。
在某些实施方案中，所述药学上可接受的辅料为载体或赋形剂。
在某些实施方案中，所述载体或赋形剂选自:离子交换剂，氧化铝，硬脂酸铝，卵磷脂，血清蛋白(例如人血清蛋白)，山梨酸，山梨酸钾，饱和植物脂肪酸的部分甘油酯混合物，水，磷酸氢二钠，磷酸氢钾，氯化钠，锌盐，胶态氧化硅，三硅酸镁，聚乙烯吡咯烷酮，纤维素物质，聚乙二醇，羧甲基纤维素钠，聚丙烯酸酯，蜂蜡，甘油，明胶，淀粉，聚乙烯-聚氧丙烯嵌段聚合物和羊毛脂。
在某些实施方案中，所述辅料为基质，例如软膏剂基质，例如油脂性基质、水溶性基质和乳剂型基质的一种或多种。
在某些实施方案中，所述软膏剂基质为水溶性基质，例如选自甘油明胶、淀粉甘油、 纤维素衍生物类、聚乙二醇类的一种或多种。在某些实施方案中，所述甘油明胶由10％-30％甘油、10％-15％明胶以及水经混合加热制成。在某些实施方案中，所述淀粉甘油由7％-10％的淀粉、70％甘油与水经混合加热制成。
可以将本发明的药物组合物制成药学上可接受的任一剂型。在某些实施方案中，本发明的药物或药物组合物被制成经皮给药制剂(例如软膏剂、硬膏剂、贴剂、涂剂、气雾剂等)或静脉注射制剂(例如液体注射剂、注射用粉剂或注射用片剂)。
在一个方面，本发明提供了一种预防或治疗银屑病的方法，包括向有此需要的受试者施用如上任一项定义的聚合物、包含所述聚合物的颗粒、聚合物胶束或药物组合物。
在某些实施方案中，所述方法包括：将所述聚合物、颗粒、聚合物胶束或药物组合物通过经皮给药的方式施予所述受试者(例如以50mg/kg-250mg/kg的剂量施予受试者)。
在某些实施方案中，所述方法包括：将所述聚合物、颗粒、聚合物胶束或药物组合物通过静脉注射的方式施予所述受试者(例如以1mg/kg-6.25mg/kg的剂量施予受试者)。
在某些实施方案中，所述方法包括：将所述聚合物、颗粒、聚合物胶束或药物组合物以每天一次的方式施用，连续给药5-7天。
在一个方面，本发明提供了如上任一项定义的聚合物、颗粒、聚合物胶束或药物组合物，其用于预防或治疗银屑病。
在某些实施方案中，所述聚合物、颗粒、聚合物胶束或药物组合物通过经皮给药的方式被施予所述受试者(例如以50mg/kg-250mg/kg的剂量施予受试者)。
在某些实施方案中，所述聚合物、颗粒、聚合物胶束或药物组合物通过静脉注射的方式被施予所述受试者(例如以1mg/kg-6.25mg/kg的剂量施予受试者)。
在某些实施方案中，所述聚合物、颗粒、聚合物胶束或药物组合物以每天一次的方式施用，连续给药5-7天。
本发明中，所述聚合物、颗粒或聚合物胶束可以作为单一有效成分施用于受试者，也可以与其他治疗银屑病的活性成分共同施用。
本发明中，所述受试者优选哺乳动物，例如牛科动物、马科动物、猪科动物、犬科动物、猫科动物、啮齿类动物、灵长类动物，例如，人。
在本发明中，除非另有说明，否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，本文中所涉及的实验室操作步骤均为相应领域内广泛使用的常规步骤。同时，为了更好地理解本发明，下面提供相关术语的定义和解释。
如本文中使用的，术语“嵌段共聚物”是指由两种或两种以上结构不同的链段组成的线性聚合物。
术语“阳离子型两亲性嵌段共聚物”是指(主要)由亲水性链段和疏水性链段组成的带有正电荷或者在水中电离后携带正电荷的共聚物。
如本文中使用的，术语“颗粒”是指以离散颗粒、丸粒、珠粒或团粒存在为特征的物质状态，而不管其大小、形状或形态如何。
如本文中使用的，术语“纳米颗粒”和“微米颗粒”分别指粒径在纳米或微米级别的颗粒，例如粒径为1nm-1000nm或1μm-1000μm的颗粒，可以是实心颗粒，也可以是胶束或囊泡等形式的颗粒。
如本文中使用的，术语“聚合物胶束”是指两亲性聚合物(例如两亲性嵌段共聚物)在溶剂(例如，水)中，当其浓度超过某一临界值时，由于疏溶剂部分(例如，疏水部分)或亲溶剂部分(例如，亲水部分)相互吸引，而形成的具有实心核-壳结构的组合体，其核由分子的疏溶剂部分(例如，疏水部分)组成，其壳由分子的亲溶剂部分(例如，亲水部分)组成。可通过例如，溶液自组装的方法，形成聚合物胶束。
如本文中使用的，术语“粒径”是指“等效粒径”，是指当被测颗粒的某种物理特性或物理行为与某一直径的同质球体(或组合)最相近时，就把该球体的直径(或组合)作为被测颗粒的等效粒径(或粒度分布)。
如本文中使用的，术语“药学上可接受的辅料”是指在生产药品和调配处方时使用的，除活性成分以外，在安全性方面已进行了合理的评估的，包含在药物制剂中的物质，其可以具有赋形、充当载体、提高稳定性、增溶、助溶和缓控释等功能中的一种或多种。
术语“聚季铵盐”是指全部或部分重复单元上带有季铵盐结构的聚合物，通常包括半合成类聚季铵盐(例如天然高分子(例如纤维素)经过季铵化改性得到的聚合物)，以及合成类聚季铵盐(例如含有双键的阳离子单体发生均聚或与其他单体发生共聚而合成的聚合物)。
有益效果
本发明发现了，阳离子型聚合物可以减少免疫细胞的激活，降低细胞因子的浓度，对银屑病有明显的治疗效果。本发明还制备了包含阳离子型两亲性聚合物的胶束，所述胶束对银屑病有明显的治疗效果。
下面将结合附图和实施例对本发明的实施方案进行详细描述，但是，本领域技术人员将理解，下列附图和实施例仅用于说明本发明，而不是对本发明的范围的限定。根据附图和优选实施方案的下列详细描述，本发明的各种目的和有利方面对于本领域技术人员来说将变得显然。
附图说明
图1示例性地描述了阳离子聚合物或其颗粒治疗银屑病的机理和过程，如图所示，阳离子聚合物可通过包括但不限于滴涂的经皮给药方式进入到患处皮肤，减少免疫细胞的激活，降低细胞因子的浓度，缓解银屑病的症状，起到治疗作用。
图2显示了两亲性嵌段共聚物PLGA-b-PDMA 474的结构式(a)、 1HNMR谱图(b)及GPC流出时间曲线(c)(均以PDMA 480作为参照)。
图3显示了阳离子型聚合物胶束cNP的粒径分布图(a)以及TEM照片(b)。
图4显示了经皮给药的体内药效实验中，各组小鼠的背部皮肤PASI评分结果。
图5显示了经皮给药的体内药效实验中，各组小鼠的背部皮肤病理切片H&E的染色结果。
图6显示了静脉注射给药的体内药效实验中，各组小鼠的背部皮肤PASI评分结果。
图7显示了静脉注射给药的体内药效实验中，各组小鼠的背部皮肤病理切片H&E的染色结果。
图8显示了体内药效实验中，各组小鼠经过治疗后后背皮肤中各种免疫因子的含量。
图9显示了cNP经皮给药的体内药效实验中，银屑病食蟹猴的背部皮肤PASI评分结果。
图10显示了食蟹猴银屑病皮肤在第0天、以及8天到第13天的照片。
图11显示了cNP经皮给药的体内药效实验中，银屑病食蟹猴的背部皮肤在第13 天即治疗五天后的病理切片H&E染色结果。
图12显示了各组小鼠肝功能和肾功能检测的结果。
图13显示了各组小鼠心、肝、脾、肺、肾的病理切片H&E染色结果。
图14显示了涂抹cNP涂剂前和涂抹cNP涂剂7天后食蟹猴肝功能和肾功能的变化情况。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例1制备两亲性嵌段共聚物PLGA-b-PDMA 474及包含其的阳离子型聚合物胶束cNP
本实施例中用于制备胶束的阳离子型两亲性嵌段共聚物为PLGA-b-PDMA 474，其结构式如下所示：
Figure PCTCN2019098604-appb-000001
1、PLGA-b-PDMA 474的制备过程如下：
S1.向9g酯封端的PLGA(市售；M η＝8000；以 1HNMR计算的M n＝12000，摩尔量为0.75mmol)中加入90mL甲苯，共沸除水；
S2.将除完水后的酯封端的PLGA冷却至室温，加入50mL CH 2Cl 2，0.08mL三乙胺(1.28mmol)；取0.06mL 2-溴代异丁酰溴(1.28mmol)，用2.5mL CH 2Cl 2稀释，冰水浴冷却下，逐滴滴加到盛有PLGA和三乙胺的烧瓶中，滴完后4小时后撤走冰浴，反应48个小时；
S3.将S2得到的产物用1mol/L的稀盐酸萃取一次，用1mol/L的碳酸氢钠萃取两次，用饱和氯化钠溶液萃取1次，有机相用无水MgSO 4干燥后，旋蒸至20mL后 在200mL冷乙醚与冷甲醇的混合沉淀剂中沉淀三次，沉淀物在真空烘箱内烘干，得到大分子引发剂PLGA-Br；
S4.将1g的大分子引发剂PLGA-Br(0.083mmol)与DMA、PMDETA、CuBr按照摩尔比1:530:1:1进行ATRP反应，加入3.5mL甲苯，冷冻-抽真空-通氮气循环三次后封管，在70℃下无水无氧反应53h；
S5.将S4得到的产物用40mL四氢呋喃稀释后过碱性氧化铝柱子除铜，并加入500mL四氢呋喃继续洗脱，收集洗脱液并旋转蒸发至15mL，滴入250mL冷的正己烷中沉淀三次，将沉淀物放入真空烘箱内烘干得到阳离子型两亲性嵌段共聚物PLGA-b-PDMA 474，数均分子量为86000，聚合物分散性指数(PDI)为1.22，DMA单体转化率为89％。
图2显示了PLGA-b-PDMA 474的核磁共振氢谱( 1HNMR)谱图和凝胶渗透色谱(GPC)流出时间曲线(均以PDMA 470为参照，PDMA 470制备过程参见实施例2)。如图2b所示，相对于PDMA 480，PLGA-b-PDMA 474的H 1NMR谱图上出现了PLGA的信号；如图2c所示，相对于PDMA 480，PLGA-b-PDMA 474的分子量有明显增加。上述结果证明了嵌段共聚物的形成。
2、基于PLGA-b-PDMA 474的阳离子型聚合物胶束(cNP)的制备过程如下：
S1.将50mg PLGA-b-PDMA 474用4mL四氢呋喃溶解，配成12.5mg/mL的聚合物溶液A；
S2.在超声破碎仪的超声作用下，将溶液A逐滴缓慢加入到pH＝3的20mL的稀盐酸水溶液中，自组装得到2.5mg/mL的PLGA-b-PDMA 474胶束溶液B；
S3.将溶液B装入截留分子量为3500的透析袋，在pH＝3的稀盐酸中透析24h；
S4.收集透析后的B溶液，往里面加入磷酸二氢钾、磷酸氢二钠、氢氧化钠并调节pH至7.4得溶液C；
S5.将溶液C加到截留量为100K的超滤离心管中，用4000g/min的转速在4℃下离心30min，浓缩到15-30mg/mL；
制备得到的阳离子型聚合物胶束cNP平均粒径为44nm，Zeta电位为+18mV。
图3a为cNP通过动态光散射(DLS)测得的粒径分布图，图3b为cNP的透射电镜(TEM)照片。如图所示，cNP为球形胶束，粒径分布较为均一。
3、基于cNP的涂剂制备过程如下：
将含有cNP的PBS的溶液超滤离心浓缩至50mg/mL，之后，与含有30％的明胶及20％的甘油的PBS溶液以体积比1:1混合，得到含有25mg/mL cNP、15％明胶和10％甘油的具有一定粘度的cNP涂剂。
实施例2
参照文献Macromolecules 31,5167-5169(1998)的方法制备水溶性阳离子聚合物PDMA 480，其结构式如下所示：
Figure PCTCN2019098604-appb-000002
核磁共振氢谱( 1HNMR)谱图和凝胶渗透色谱(GPC)流出时间曲线见图2。
参照实施例1的制备方法，制备基于PDMA的涂剂。
实施例3
PAMAM 3代树枝状聚合物(PAMAM-G3)，分子量为6900，于Sigma购买并后处理得到(货品名为PAMAM dendrimer,ethylenediamine core,generation 3.0 solution 20wt.％in methanol)，购买后的聚合物溶液于纯水中用截留分子量为3500的透析袋透析24小时后冻干得到PAMAM-G3固体。参照实施例1的制备方法，制备基于PAMAM-G3的涂剂。
实施例4
海地美溴铵(hexadimethrine bromide，HDMBr)，于阿拉丁购买得到。参照实施例1的制备方法，制备基于海地美溴铵的涂剂。
实施例5
鱼精蛋白，于阿拉丁购买得到。参照实施例1的制备方法，制备基于鱼精蛋白的涂剂。
实施例6
聚季铵盐-10，于上海安研商贸有限公司购买得到。参照实施例1的制备方法，制备基于聚季铵盐-10的涂剂。
实施例7
聚赖氨酸(PLL)参照文献J.Am.Chem.Soc.1999,121,5919-5929的方法制备，结构如下：
Figure PCTCN2019098604-appb-000003
参照实施例1的制备方法，制备基于聚赖氨酸的涂剂。
实验例1阳离子聚合物或阳离子型聚合物胶束对银屑病的体内药效实验
实验(1)：阳离子聚合物或阳离子型聚合物胶束对银屑病小鼠的体内药效实验
实验过程：
1、分组：将体重为20g的Bal B/C小鼠，分为1组模型组、1组空白组、两组静脉注射组(即，PDMA(i.v.)给药组和cNP(i.v.)给药组)，八组经皮给药组(即，PDMA涂剂给药组、cNP涂剂给药组、鱼精蛋白涂剂给药组、聚季铵盐涂剂给药组、海地美溴铵涂剂给药组、PAMAM-G3涂剂给药组和聚赖氨酸涂剂给药组)，每组各10只。
2、造模：小鼠用异氟烷气体麻醉后，第一天先用剃毛刀剃掉背部2cm*2cm皮肤的毛发，并连续七天在这块皮肤上涂抹62.5mg艾达乐的咪喹莫特乳膏至吸收，以诱导小鼠银屑病样皮损，涂抹七天后银屑病模型建成。
3、给药：
经皮给药：在第8天成模型时，即银屑病小鼠出现红肿和/或鳞屑等症状时，分别以250mg/kg即1.25mg/cm 2将不同涂剂涂抹在对应组别的小鼠背部皮肤上，并敷上纱布，以绷带缠绕来固定。连续对小鼠经皮给药5天；对模型组小鼠在背部皮肤涂抹甘油明胶基底作为对照。
静脉给药：在第8天成模型时，即银屑病小鼠出现红肿和/或鳞屑等症状时，分别以剂量12.5mg/kg将cNP胶束的PBS溶液或PDMA 480的PBS溶液通过尾静脉注射到小鼠体内；连续对小鼠通过尾静脉注射给药5天；对模型组小鼠通过尾静脉注射等体 积的PBS作为对照。
空白组小鼠不做任何处理。
4、记录PASI评分：在给药期间，每一天用肉眼观测小鼠背部皮肤状况，根据文献J Immunol 2009；182:5836-5845的评分标准分别对红肿程度、鳞屑数量、背部皮肤厚度进行评分，分数为0-4分：0分：无；1分：情况轻微；2分：情况中等；3分：情况严重；4分：情况非常严重。并由这三项的分数的总分得到总的评分。
5、采血与组织切片：在第13天即治疗五天后采血取血清测量测肝功肾功，并安乐死小鼠，取其皮肤、心、肝、脾、肺、肾做组织切片研究，并测量细胞因子含量。
细胞因子的含量的测量方法：
对第13天处死小鼠取得的皮肤取部分浸泡于TRIzol试剂中，将其用分散机研磨成匀浆后，使用Rneasy-universal Tissue Kit试剂盒提取RNA，将RNA逆转录为cDNA后用RT-PCR方法对TNF-α、IL-6、IL-17A、IL17-R、IL22、IL23的mRNA水平进行半定量分析。
实验结果：
图4显示了经皮给药的体内药效实验中，各组小鼠的背部皮肤PASI评分结果。其中：
图4a显示了空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组，鱼精蛋白涂剂给药组，聚季铵盐涂剂给药组，海地美溴铵涂剂给药组，PAMAM-G3涂剂给药组，聚赖氨酸涂剂给药组在第1天至第13天的后背皮肤红肿程度得分；
图4b显示了空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组，鱼精蛋白涂剂给药组，聚季铵盐涂剂给药组，海地美溴铵涂剂给药组，PAMAM-G3涂剂给药组，聚赖氨酸涂剂给药组在第1天至第13天的后背皮肤鳞屑程度得分；
图4c显示了空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组，鱼精蛋白涂剂给药组，聚季铵盐涂剂给药组，海地美溴铵涂剂给药组，PAMAM-G3涂剂给药组，聚赖氨酸涂剂给药组在第1天至第13天的后背皮肤厚度程度得分；
图4d显示了空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组，鱼精蛋白涂剂给药组，聚季铵盐涂剂给药组，海地美溴铵涂剂给药组，PAMAM-G3涂剂给药组，聚赖氨酸涂剂给药组在第1天至第13天的后背皮肤总得分。
图5显示了经皮给药的体内药效实验中，各组小鼠的背部皮肤病理切片H&E的 染色结果，包括空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组，鱼精蛋白涂剂给药组，聚季铵盐涂剂给药组，海地美溴铵涂剂给药组，PAMAM-G3涂剂给药组，聚赖氨酸涂剂给药组的H&E染色结果。
图6显示了静脉注射给药的体内药效实验中，各组小鼠的背部皮肤PASI评分结果，其中:
图6a显示了空白组、模型组、PDMA(i.v.)给药组，cNP(i.v.)给药组在第1天至第13天的后被皮肤红肿程度得分；
图6b显示了空白组、模型组、PDMA(i.v.)给药组，cNP(i.v.)给药组在第1天至第13天的后背皮肤鳞屑程度得分；
图6c显示了空白组、模型组、PDMA(i.v.)给药组，cNP(i.v.)给药组在第1天至第13天的后背皮肤厚度程度得分；
图6d显示了空白组、模型组、PDMA(i.v.)给药组，cNP(i.v.)给药组在第1天至第13天的后背皮肤总得分。
图7显示了静脉注射给药的体内药效实验中，各组小鼠的背部皮肤病理切片H&E的染色结果，包括空白组、模型组、PDMA(i.v.)给药组，cNP(i.v.)给药组的染色结果。
图8显示了体内药效实验中，空白组、模型组、PDMA涂剂给药组，cNP涂剂给药组、PDMA(i.v.)给药组，cNP(i.v.)给药组在第13天后背皮肤中TNF-α、IL-6、IL-17A、IL-17F、IL-22、IL-23A免疫因子的mRNA水平。
综合图4-图8的结果，可以看出，不管是用阳离子聚合物涂剂或阳离子聚合物胶束涂剂涂抹皮肤治疗，还是将阳离子聚合物或其胶束静脉注射治疗，都能对银屑病小鼠起到一定的治疗效果，表现为相对于模型组来说，给药组皮肤的红肿情况有所缓解，鳞屑减少，角质化程度减少，炎症细胞浸润减少，表皮和真皮的厚度更接近于正常组，棘层增生的情况得到改善。治疗五天以后，PDMA涂剂给药组，cNP涂剂给药组、PDMA(i.v.)给药组，cNP(i.v.)给药组TNF-α、IL-6、IL-17A、IL-17F、IL-22、IL-23A免疫因子的mRNA水平相对于模型组有一定下降，说明有效地抑制了银屑病的炎症反应。
实验(2)：阳离子聚合物胶束涂剂对银屑病食蟹猴的体内药效实验
由于不同种属之间皮肤组织结构差异较大，实验(2)拟选用灵长类动物食蟹猴进行实验，以更好地评价阳离子聚合物或其胶束的治疗效果。食蟹猴代谢、功能和结构等生物学特性与人类相似，因此是研究人类相关疾病的较佳模式动物。以食蟹猴作 为实验动物对象进行动物实验，可以更真实有效地评价阳离子聚合物或其胶束的实际药效，评估在实际人类疾病治疗的可行性。
实验过程：
1、分组：食蟹猴3只，雄性，体重约为4-5kg，饲养周期为8周。
2、造模：每只食蟹猴剃毛后使用咪喹莫特乳膏约1g分别涂抹动物的后背上下两侧各2cm*2cm皮肤进行造模，涂至皮肤基本吸收，连续涂抹7天以后模型成立。
3、给药：
对每只食蟹猴的下侧发炎皮肤连续5天涂抹2mg/kg即2.5mg/cm 2含有cNP的甘油明胶涂剂0.1mL进行治疗，每天涂抹两次，并对上侧发炎皮肤涂抹0.21mL无cNP的甘油明胶基底进行自身对照(模型皮肤)，每天涂抹两次。发炎皮肤采用PASI评分并拍照记录。在治疗终点即鳞屑不再出现或不再红肿时，对动物用“舒泰50”全身麻醉后剪下相应皮肤部位的0.5cm*1cm皮肤做病理切片并缝合创口。另取一块0.5cm*1cm正常部位皮肤作为空白对照。
实验结果：
图9显示了cNP经皮给药的体内药效实验中，银屑病食蟹猴的背部皮肤PASI评分结果。其中：
图9a显示了空白组、模型组、cNP涂剂给药组在第1天至第13天的后背皮肤红肿程度得分；
图9b显示了空白组、模型组、cNP涂剂给药组在第1天至第13天的后背皮肤鳞屑程度得分；
图9c显示了空白组、模型组、cNP涂剂给药组在第1天至第13天的后背皮肤厚度程度得分；
图9d显示了空白组、模型组、cNP涂剂给药组在第1天至第13天的后背皮肤总得分。
图10显示了食蟹猴银屑病皮肤在第0天、第8天到第13天的照片。其中三只食蟹猴(编号0126、0148和0150)背部下侧cNP治疗的银屑病皮肤以及上侧自身对照的模型皮肤的照片在第0天、第8天到第13天的照片。其中第0天的照片是造模前的皮肤照片，第8天是模型建立但没开始给药的皮肤照片，第9天到第13天是给药后的皮肤照片。黄色虚线方框为造模时选定的2cm*2cm的范围。
图11显示了cNP经皮给药的体内药效实验中，银屑病食蟹猴的背部皮肤在第13天即治疗五天后的病理切片H&E染色结果，包括空白组(正常猴子的皮肤)、模型组(银屑病猴子上侧没涂cNP的皮肤)、cNP涂剂给药组(银屑病猴子下侧涂cNP的皮肤)的H&E染色结果。其中大图为放大4倍的的图片，小图为放大20倍的图片。
综合图9-图11的结果，可以看出，用cNP涂剂涂抹皮肤治疗银屑病对银屑病食蟹猴能起到一定的治疗效果，表现为相对于自身对照的模型组皮肤来说来说，cNP给药后的皮肤红肿有消退，鳞屑减少并且没有新鳞屑出现，而模型组的皮肤随着时间延长鳞屑不断增加，皮肤越来越厚且硬。从H&E染色结果来看，cNP治疗后的皮肤角质化程度减少，角化不全程度下降，表皮和真皮的厚度比模型组的要薄。说明cNP涂剂能有效渗入灵长类的皮肤，并能有效地抑制银屑病的炎症反应从而缓解病情。
实验例2阳离子聚合物或阳离子型聚合物胶束的体内安全性评价
实验(1)：阳离子聚合物或阳离子型聚合物胶束在小鼠体内安全性评价
实验过程见实验例1中实验(1)的过程，在第13天对空白组模型组、PDMA涂剂给药组、cNP涂剂给药组、PDMA(i.v.)给药组、cNP(i.v.)给药组进行眼眶取血，离心血液取上层血清，用全自动生化分析仪测量反映肝功能的碱性磷酸酶(ALP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)指标及反映肾功能的尿酸、尿素、肌酐指标。另外在第13天安乐死各组小鼠，取心、肝、脾、肺、脏做石蜡切片并进行H&E染色。
实验(2)：阳离子型聚合物胶束在食蟹猴体内安全性评价
实验过程：本实验选取3只食蟹猴，先用剃毛刀在动物的后背剃掉2cm*2cm面积的毛发，再于皮肤上涂抹0.2mL阳离子聚合物胶束明胶溶液，连续7天给药，于第8天进行静脉取血，分离血清，用全自动生化分析仪测量肝功能的碱性磷酸酶(ALP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)指标及反映肾功能的尿酸、尿素、肌酐指标。
实验结果：
图12显示了各组小鼠肝功能和肾功能检测的结果。如图所示，cNP给药组不管是静脉注射给药还是经皮给药的肝功能和肾功能都更接近于正常组的大鼠，而PDMA组则离正常组小鼠的指标有一定差距，如PDMA涂剂组AST、ALT、尿酸、尿素都偏高。结果说明cNP比PDMA的肝毒性、肾毒性小。
图13显示了各组小鼠心、肝、脾、肺、肾的病理切片H&E染色结果。如图所示，cNP给药组不管是静脉注射给药还是经皮给药的都对内脏没有明显损伤，而PDMA注射组则引起了肝细胞的凋亡、肺泡出血、肾小球萎缩等症状，PDMA经皮给药组引起了肝细胞水肿、肾小球萎缩等症状，结果说明cNP比PDMA对内脏的毒性小。
图14显示了涂抹cNP涂剂前和涂抹cNP涂剂7天后食蟹猴肝功能和肾功能的变化情况。如图所示，食蟹猴在涂抹cNP涂剂7天以后与自身给药前对比，ALP、AST、ALT、肌酐、尿酸、尿素都没有太大的变化，说明cNP涂剂经皮给药没有明显体内毒性，并不会影响正常的肝功能和肾功能。
尽管本发明的具体实施方式已经得到详细的描述，但本领域技术人员将理解：根据已经公开的所有教导，可以对细节进行各种修改和变动，并且这些改变均在本发明的保护范围之内。本发明的全部范围由所附权利要求及其任何等同物给出。

Claims (10)

  1. 聚合物或包含所述聚合物的颗粒作为有效成分用于制备药物的用途，所述药物用于预防或治疗银屑病，所述聚合物选自以下阳离子聚合物中的一种或多种：聚甲基丙烯酸2-(二甲氨基)乙酯、聚α-氨基戊内酯、聚醚酰亚胺、聚酰胺-胺型树枝状聚合物、聚(2-二甲氨基乙基硫)己内酯、聚赖氨酸、阳离子淀粉、聚阳离子型Q-葡聚糖、聚季铵盐、聚丙烯酰胺、鱼精蛋白、海地美溴铵，或者，所述聚合物为阳离子型共聚物，其包含以上任意一种或多种阳离子聚合物的重复单元；
    优选地，所述阳离子型共聚物为阳离子型两亲性共聚物；
    优选地，所述阳离子型共聚物为阳离子型嵌段共聚物；
    优选地，所述阳离子型嵌段共聚物包含电中性聚合物链段和以上任意一种或多种阳离子聚合物的链段；
    优选地，所述电中性聚合物链段选自以下一种或多种聚合物的链段：聚乳酸-羟基乙酸无规共聚物、疏水性聚磷酸酯、疏水性聚碳酸酯、聚乙二醇、聚己内酯；
    优选地，所述阳离子型嵌段共聚物为阳离子型两亲性嵌段共聚物，其包含亲水段和疏水段，其中，所述亲水段为阳离子聚合物的链段，所述疏水段为电中性聚合物链段；
    优选地，所述阳离子淀粉选自：2-羟基-3-(三甲基氨根)丙基醚淀粉氯化物；
    优选地，所述聚季铵盐选自：聚季铵盐-1、聚季铵盐-2、聚季铵盐-5、聚季铵盐-6、聚季铵盐-7、聚季铵盐-10、聚季铵盐-11、聚季铵盐-12、聚季铵盐-13、聚季铵盐-14、聚季铵盐-16、聚季铵盐-17、聚季铵盐-19、聚季铵盐-20、聚季铵盐-22、聚季铵盐-27、聚季铵盐-28、聚季铵盐-29、聚季铵盐-30、聚季铵盐-32、聚季铵盐-37、聚季铵盐-38、聚季铵盐-39、聚季铵盐-44、聚季铵盐-46、聚季铵盐-47、聚季铵盐-51、聚季铵盐-53、聚季铵盐-55、聚季铵盐-58、聚季铵盐-60、聚季铵盐-67。
  2. 权利要求1的用途，所述阳离子聚合物或其链段具有以下特征中的一个或多个：
    (1)所述阳离子聚合物或其链段具有200-500、500-5k或5k-500k的数均分子量；
    (2)所述阳离子聚合物或其链段具有200-500、500-5k或5k-500k的重均分子量；
    (3)所述阳离子聚合物或其链段具有大于1且小于1.5的聚合物分散性指数。
  3. 权利要求1或2的用途，所述电中性聚合物链段具有以下特征中的一个或多个：
    (1)所述电中性聚合物链段具有5k-500k的数均分子量；
    (2)所述电中性聚合物链段具有5k-500k的重均分子量；
    (3)所述电中性聚合物链段具有大于1且小于1.5的聚合物分散性指数。
  4. 权利要求1-3任一项的用途，所述聚合物为阳离子型两亲性嵌段共聚物，其中，亲水的阳离子聚合物链段为PDMA链段，疏水的电中性链段为PLGA链段；优选地，所述聚合物具有以下特征中的一个或多个：
    (1)PDMA链段的数均分子量为30k-80k；
    (2)PLGA链段的数均分子量为8k-10k；
    (3)PLGA链段中，乳酸结构单元和羟基乙酸结构单元的个数各自独立地选自80-100的正整数。
  5. 权利要求1-4任一项的用途，所述药物被制成药学上可接受的剂型，例如经皮给药制剂或静脉注射制剂。
  6. 一种聚合物胶束，其包含权利要求1-4任一项定义的阳离子型两亲性共聚物，或由权利要求1-4任一项定义的阳离子型两亲性共聚物组成；优选地，所述聚合物胶束具有以下特征中的一个或多个：
    (1)所述聚合物胶束具有30nm-200nm的粒径；
    (2)所述聚合物胶束的粒径具有0.1-0.3的多分散指数；
    (3)所述聚合物胶束具有+15mV至+20mV的Zeta电位；
    (4)所述聚合物胶束为球形胶束。
  7. 制备权利要求6的聚合物胶束的方法，包括以下步骤：
    步骤1：提供权利要求1-4任一项定义的阳离子型两亲性共聚物；
    步骤2：将所述阳离子型两亲性共聚物在溶液中进行自组装，形成所述聚合物胶束。
  8. 一种组合物，其包含权利要求6的聚合物胶束；
    优选地，所述组合物还包含溶剂，例如水。
  9. 权利要求6的聚合物胶束在制备药物中的用途，所述药物用于预防或治疗受试者的银屑病。
  10. 一种药物组合物，其包含权利要求6的聚合物胶束，以及一种或多种药学上可接受的辅料。
PCT/CN2019/098604 2019-07-31 2019-07-31 可用于治疗银屑病的阳离子聚合物及颗粒 WO2021016924A1 (zh)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040009503A1 (en) * 2002-07-03 2004-01-15 Molecular Staging, Inc. Immune modulatory activity of human ribonucleases
CN101265312A (zh) * 2008-05-07 2008-09-17 天津大学 两亲性三嵌段共聚物及制备方法和应用
CN102639142A (zh) * 2009-09-16 2012-08-15 杜克大学 内体Toll样受体活化的抑制
US20120328544A1 (en) * 2003-12-22 2012-12-27 Nevada Naturals, Inc. Dermatological Treatment Methods And Formulations
CN110179749A (zh) * 2018-04-27 2019-08-30 中山大学 可用于治疗类风湿性关节炎的聚合物纳米颗粒

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040009503A1 (en) * 2002-07-03 2004-01-15 Molecular Staging, Inc. Immune modulatory activity of human ribonucleases
US20120328544A1 (en) * 2003-12-22 2012-12-27 Nevada Naturals, Inc. Dermatological Treatment Methods And Formulations
CN101265312A (zh) * 2008-05-07 2008-09-17 天津大学 两亲性三嵌段共聚物及制备方法和应用
CN102639142A (zh) * 2009-09-16 2012-08-15 杜克大学 内体Toll样受体活化的抑制
CN110179749A (zh) * 2018-04-27 2019-08-30 中山大学 可用于治疗类风湿性关节炎的聚合物纳米颗粒

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