EP3710061A1 - Polymer-prodrugs und subkutane und / oder intramuskuläre verabreichung davon - Google Patents

Polymer-prodrugs und subkutane und / oder intramuskuläre verabreichung davon

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Publication number
EP3710061A1
EP3710061A1 EP18800227.3A EP18800227A EP3710061A1 EP 3710061 A1 EP3710061 A1 EP 3710061A1 EP 18800227 A EP18800227 A EP 18800227A EP 3710061 A1 EP3710061 A1 EP 3710061A1
Authority
EP
European Patent Office
Prior art keywords
polymer
molecule
radical polymerization
active molecule
polymeric
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.)
Pending
Application number
EP18800227.3A
Other languages
English (en)
French (fr)
Inventor
Nicolas Tsapis
Julien Nicolas
Tanguy BOISSENOT
Alexandre BORDAT
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.)
Centre National de la Recherche Scientifique CNRS
Universite Paris Saclay
Original Assignee
Centre National de la Recherche Scientifique CNRS
Universite Paris Sud Paris 11
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 Centre National de la Recherche Scientifique CNRS, Universite Paris Sud Paris 11 filed Critical Centre National de la Recherche Scientifique CNRS
Publication of EP3710061A1 publication Critical patent/EP3710061A1/de
Pending legal-status Critical Current

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Classifications

    • 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
    • A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/58—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F220/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride ester, amide, imide or nitrile thereof
    • C08F220/02—Monocarboxylic acids having less than ten carbon atoms; Derivatives thereof
    • C08F220/52—Amides or imides
    • C08F220/54—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide
    • C08F220/56—Acrylamide; Methacrylamide
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F293/00—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule
    • C08F293/005—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F2438/00—Living radical polymerisation
    • C08F2438/03—Use of a di- or tri-thiocarbonylthio compound, e.g. di- or tri-thioester, di- or tri-thiocarbamate, or a xanthate as chain transfer agent, e.g . Reversible Addition Fragmentation chain Transfer [RAFT] or Macromolecular Design via Interchange of Xanthates [MADIX]

Definitions

  • the present invention relates to novel prodrugs of active molecules. These prodrugs in particular allow the subcutaneous or intramuscular administration of active molecules whose subcutaneous or intramuscular administration is problematic or impossible, particularly because of the toxicity at the injection site.
  • the prodrugs of the invention comprise an active ingredient, covalently bonded to a polymer chain, preferably a hydrophilic and / or heat-sensitive polymer chain.
  • PAs have low bioavailability through the SC pathway. Either their physico-chemical properties (log P, solubility, molar mass, etc.) do not allow them to cross the SC barrier, or they are not stable in a biological environment and are degraded before reaching the circulation, and sometimes even they combine the two disadvantages;
  • SC subcutaneous route
  • IV intravenous
  • Otsuka in collaboration with BMS has developed a technology based on cyclodextrins. These cyclic oligosaccharides have a hydrophobic inner cavity and a hydrophilic outer surface. The hydrophobic PAs will thus encapsulate in the internal cavity, thus inducing an increase in their apparent solubility. This increase in solubility results in an increase in SC bioavailability.
  • Adocia is developing bio-chaperone polymers that will be associated with biomolecules such as insulin through physicochemical interactions. Insulin will be stabilized in individualized form ("monomeric") and will have a higher SC bioavailability than insulin in its classic hexameric form (faster absorption of the "monomer” form because its molar mass is lower). We play here on the stability of the insulin monomer at high concentration.
  • Halozyme develops, for its part, a formulation based on Hyaluronidase; an enzyme that will reversibly degrade SC tissue predominantly composed of hyaluronic acid.
  • the volume of direct injection will therefore be able to be increased from 2 to 5 mL
  • the administration of a high concentration PA to reach effective therapeutic doses for example from 5 mg / ml and in particular from 10 mg / ml, generally has the technical disadvantage that the formulation becomes too viscous and incompatible with SC injection.
  • the present invention aims to solve the technical problems stated above.
  • the object of the present invention is to solve the technical problem of providing an injectable composition, in particular by the subcutaneous (“SC”) or intramuscular (“IM”) route of one or more active ingredients, for example anticancer, avoiding problems of bioavailability and stability of the active ingredient and not causing irritation or necrosis, to the patient at the injection site.
  • SC subcutaneous
  • IM intramuscular
  • the present invention aims to solve the technical problem of providing an injectable composition, particularly subcutaneously (“SC”) or intramuscular (“IM”), with a high concentration of PA, and in particular concentrations 5 mg / mL or 10 mg / mL or more.
  • SC subcutaneously
  • IM intramuscular
  • the object of the present invention is to solve the technical problem of providing an injectable composition, in particular subcutaneously (“SC”) or intramuscular (“IM”), of a therapeutically effective amount of at least one a pharmaceutically active ingredient in a small volume of solution for injection, for example from 1 to 20 ml.
  • SC subcutaneously
  • IM intramuscular
  • the object of the present invention is to solve the technical problem of providing an injectable composition, in particular by the subcutaneous (“SC”) or intramuscular (“IM”) method, of at least one pharmaceutically active principle formulated in a solution. not very viscous, allowing easy injection.
  • SC subcutaneous
  • IM intramuscular
  • the inventors have developed a new polymer prodrug technology that allows the administration of the active principles by SC or IM without skin / local toxicity observed.
  • the prodrug approach makes it possible to inactivate the PA of interest at SC or IM tissue and to release it by cleaving the PA / polymer bond once in the general circulation or at the site. 'action.
  • the chemical coupling between PAs and very water-soluble polymers such as polyacrylamide also makes it possible to modify the physico-chemical properties of the therapeutic molecules. The physicochemical properties of the polymer will be conferred on the PA.
  • polyacrylamide makes it possible to increase the solubility of PA very even when it has a low molecular weight. These characteristics make it possible to maintain a high concentration stability while limiting the viscosity, to maximize its absorption from SC or IM tissue and to limit its metabolism at this level, thus leading to increased bioavailability.
  • the polymeric prodrugs of the invention increase the bioavailability and stability of the AP.
  • the polymeric prodrugs of the invention maintain a high concentration of PA stability, and in particular at a concentration of at least 1 mg / mL, for example at least 2 mg / mL and preferably at least 5 mg / mL in equivalent concentration of AP.
  • the polymeric prodrug is injectable at an equivalent PA concentration of at least 10 mg / mL and preferably at least 20 mg / mL.
  • the polymeric prodrugs of the invention maximize the absorption of AP from SC or IM tissue.
  • the polymeric prodrugs of the invention limit the metabolism of PA, thus leading to increased bioavailability.
  • the polymeric prodrugs of the invention allow the SC injection of high doses of AP while avoiding the phenomena of SC or IM irritation and / or necrosis.
  • the polymeric prodrugs of the invention make it possible to adapt to a large number of active ingredients, which makes the polymeric prodrugs, their preparation and their uses particularly interesting.
  • polymeric prodrugs whose properties allow them to be administered SC or IM without irritation or necrosis reactions.
  • the inventors have found that the physicochemical properties of the polymer are conferred on the prodrug, and in particular the solubility properties in a solution for injection by the SC or IM route.
  • polymeric prodrugs according to the present invention leads to obtaining solutions at equivalent concentrations of active ingredient, for example from 1 to more than 20 mg / ml in comparison with the low solubilities of certain active ingredients around of 1 ⁇ g / mL.
  • the polymeric prodrugs of this invention were chosen for their high solubility in relation to a large number of tested polymers. Because of their extremely hydrophilic nature, they will be able to solubilize hydrophobic PAs while having a low molecular weight. Due to this low molecular weight, the charge rate will be high, the viscosity will be low and the absorption will be rapid at the tissue level. SC (the rate of absorption is inversely proportional to the molar mass).
  • the choice of the bond between the polymer and the PA makes it possible to release the molecule only after absorption from SC or IM tissue (no early release at the tissue level). These 3 characteristics make it possible to inject large quantities of PA without toxicity.
  • in vivo assays show no evidence of toxicity, irritation or necrosis following SC administration of a cytotoxic prodrug that usually leads to this type of reaction when injected without our formulation.
  • the prodrugs used in the invention therefore show adequate properties to be administered SC or IM.
  • the present invention relates to a polymeric prodrug comprising:
  • a second pharmaceutically active molecule covalently coupled to the terminal portion of the polymer.
  • the proximal and terminal portions are arbitrarily defined as the ends of a substantially linear polymer chain, i.e. the pendant chains present are of shorter length than the main chain.
  • the main chain is the chain comprising the reactive groups for the polymerization and propagating during the polymerization.
  • the proximal and end portions denote the ends of the polymer.
  • the proximal portion refers to the non-elongated end and the end portion to the elongated end.
  • the terms "parts" proximal and terminal designate globally the proximal and terminal ends, respectively so the proximal portion may comprise the first PA and the first monomer and the terminal portion may comprise the last monomer and, if present, the second PA.
  • proximal and terminal portions for the radical polymerization control agent are also referred to in the invention.
  • the invention particularly relates to a polymeric prodrug comprising a proximal portion and an end portion and comprising:
  • At least one first pharmaceutically active molecule at least one polymer chain formed at least in part from acrylamide monomers or one of its derivatives,
  • At least one radical polymerization control agent comprising a proximal portion and an end portion; the first pharmaceutically active molecule being located in the proximal portion of the prodrug polymer and covalently bound to the proximal portion of the radical polymerization control agent,
  • the terminal portion of the radical polymerization control agent being located at the end portion of the prodrug polymer and being covalently bonded to the polymer chain.
  • the invention also relates to a water-soluble polymer prodrug comprising a proximal portion and a terminal portion and comprising: at least one first pharmaceutically active molecule, at least one polymer chain, at least one radical polymerization control agent comprising a part proximal and a terminal part; the first pharmaceutically active molecule being located at a proximal portion of the prodrug polymer and covalently bound to the proximal portion of the radical polymerization control agent, the terminal portion of the radical polymerization control agent being located at the terminal portion of the prodrug polymer and being covalently bound to the chain of polymer, said solubility being evaluated at a concentration of 200 mg / ml in distilled water with paclitaxel as the first pharmaceutically active molecule.
  • the polymer is formed at least partly of acrylamide monomer, or one of its derivatives, and of co-monomers to form random or block polymers, for example poly (acrylamide-co). acrylonitrile).
  • the polymer is a poly (acrylamide).
  • the present invention relates to a polymeric prodrug comprising:
  • a water-soluble polymer chain said polymer comprising a proximal portion and an end portion;
  • a first pharmaceutically active molecule covalently coupled to the proximal polymer
  • a second pharmaceutically active molecule covalently coupled to the terminal portion of the polymer
  • solubility being evaluated at a concentration of 200 mg / ml in distilled water with paclitaxel as the first pharmaceutically active molecule.
  • the polymer chain has a polydispersity index of less than 1.5, said polydispersity index determined by steric exclusion chromatography. According to one variant, the polymer chain has a molar mass of 1000 to 1,000,000 g / mol, preferably less than 100,000 g / mol, preferably less than 50,000 g / mol.
  • the polymer comprises a radical polymerization control agent being chosen from the control agents of the controlled radical polymerization controlled by reversible addition-fragmentation chain transfer (in English Reversible Addition-Fragmentation Chain Transfer (RAFT), radical polymerization controlled by ATOM Transfer Radical Polymerization (ATRP)) and its derivatives (controlled radical polymerization by copper (I)), nitroxide-controlled radical polymerization (Nitroxide-Mediated Polymerization (NMP), controlled radical polymerization cobalt (CoMRP), organotelluric controlled radical polymerization (TERP) and organoantimoin controlled radical polymerization (SbRP), and, for example, thiocarbonylthio transfer systems such as dithiocarbonate, xanthate, dithiocarbamate and trithiocarbonate, among the transition metal-based complexes (Cu, Fe, Ru, etc.), among the alkoxyamines, among the cobalt-based complexes, the organotellures and among the organoantimoi
  • the polymer comprises a radical polymerization control agent comprising, in the terminal part, a terminal alkyl chain, for example comprising from 1 to 20 carbon atoms, a carboxylic acid function, an alcohol function, an amine function or an amide function. a thiol function, said function optionally being supported by the terminal alkyl chain, and said function being optionally linked to a second pharmaceutically active molecule.
  • a radical polymerization control agent comprising, in the terminal part, a terminal alkyl chain, for example comprising from 1 to 20 carbon atoms, a carboxylic acid function, an alcohol function, an amine function or an amide function.
  • a thiol function said function optionally being supported by the terminal alkyl chain, and said function being optionally linked to a second pharmaceutically active molecule.
  • the polymer comprises a radical polymerization control agent comprising in part proximally a proximal function chosen from an amide, ester, carbonate, carbamate, succinate, disulfide, acetal, thioether or triazole function; and / or linker diglycolate, succinate, succinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC), glycinate, glucuronate, valine-citrulline, maleimide, said function and / or proximal linker forming a covalent bond with the first pharmaceutically acceptable principle active.
  • a radical polymerization control agent comprising in part proximally a proximal function chosen from an amide, ester, carbonate, carbamate, succinate, disulfide, acetal, thioether or triazole function; and / or linker diglycolate, succinate, succinimidyl 4- (N-maleimidomethyl
  • the active molecule is an anticancer molecule, an antibiotic molecule (bacterio / fungo-static, bacterial / fungo-toxic or antiviral agent), an antidiabetic molecule, a molecule treating vascular or cardiovascular pathologies, a molecule treating pathologies of the central nervous system, an anti-inflammatory molecule, an agonist molecule of a physiological receptor, a molecule antagonist or partially antagonist of a physiological receptor, an immunomodulatory molecule.
  • the active molecule is an anticancer molecule chosen from: paclitaxel, docetaxel, gemcitabine, cladribine, capecitabine, daunorubicin, doxorubicin, epirubicin, idarubicin, actinomycin, amsacrine , dacarbazine, dactinomycin, vincristine, vimblastine, vindesine, methotrexate, colchiccin, cyclophosphamide, azathioprine, 6-mercaptopurine, lomustine, carmustine, dacarbazine, cisplatin, fluorouracil, tenoposide or etoposide, fotemustine, mitomycin C, mitoxantrone, streptozocin, trabectedin, vinflunine, vinorelbine, asernic trioxide, bendamustine, busulfan, cabazitaxel, carboplatin, e
  • the polymeric prodrug induces a time-delayed release of the pharmaceutically active molecule into the bloodstream or at the site of action, for example at the level of a tumor or at the intracellular level.
  • the present invention relates to a polymeric prodrug according to the invention, for its use in a method of therapeutic treatment, or in a method of diagnosis, or in a medical imaging method, in a human or animal by subcutaneous administration or intramuscular.
  • the present invention relates to a polymeric prodrug according to the invention, for its use in a method of therapeutic treatment of a human or animal by subcutaneous or intramuscular administration, said polymeric prodrug comprising a covalent bond between a pharmaceutically active molecule and a polymer, said pharmaceutically active molecule not being administrable subcutaneously or intramuscularly because of its toxicity at the injection site (tissue irritation / necrosis) when not bound by covalently bonding to said polymer, preferably said polymeric prodrug exhibiting a bioavailability of the molecule preventing local toxicities (at the injection site) and releasing the pharmaceutically active molecule into the bloodstream.
  • the treatment method according to the invention is a method of treating cancer.
  • the present invention relates to a method of controlled radical polymerization, in particular by the so-called initiator active principle method, of at least one polymeric prodrug according to the invention, said process comprising the steps of:
  • the present invention also relates to a controlled radical polymerization process of at least one polymeric prodrug according to the invention, said process comprising the steps of:
  • the covalent coupling of a second pharmaceutically active molecule at the terminal end of the polymer prodrug optionally, the covalent coupling of a second pharmaceutically active molecule at the terminal end of the polymer prodrug.
  • the present invention also relates to a composition for injection into a tissue of a mammal, preferably a human being, and in particular formulated for subcutaneous or intramuscular injection, said composition comprising a polymeric prodrug as defined according to the invention. .
  • the present invention relates to a polymeric prodrug for use in a method of treatment.
  • This method comprises subcutaneous or intramuscular administration of a pharmaceutically effective amount of said polymeric prodrug to a patient.
  • Polymer refers to a polymer or copolymer.
  • radical polymerization initiator refers to all of the compounds used to produce radicals and thereby initiate radical polymerization. These compounds possess a chemical function capable of releasing radicals under the action of heat, light irradiation, oxidation-reduction reaction, ionizing radiation, electrochemical reactions and sonication.
  • Non-limiting examples of initiators include azo-type compounds, such as 2,2'-azobis (2-methylpropionitrile), 4,4'-azobis (4-cyanovaleric), 1,1 '- azobis (cyclohexanecarbonyl); inorganic peroxide type; or organic peroxide type such as benzoyl peroxide, lauroyl peroxide, methyl ethyl ketone peroxide, tert-butyl peroxybenzoate.
  • azo-type compounds such as 2,2'-azobis (2-methylpropionitrile), 4,4'-azobis (4-cyanovaleric), 1,1 '- azobis (cyclohexanecarbonyl); inorganic peroxide type; or organic peroxide type such as benzoyl peroxide, lauroyl peroxide, methyl ethyl ketone peroxide, tert-butyl peroxybenzoate.
  • control agent refers to all of the compounds used in a polymerization reaction in order to obtain polymers having a number average molecular weight ratio on weight average molar mass, or dispersity, less than 1. 5.
  • the nature of these compounds depends on the controlled radical polymerization technique implemented.
  • RAFT controlled reversible addition-fragmentation chain transfer polymerization
  • these are compounds of the dithiocarbonate (xanthate), trithiocarbonate, dithioester or dithiocarbamate type.
  • NMP nitroxide-controlled radical polymerization
  • the radical polymerization initiator and the control agent are combined into one and the same alkoxyamine molecule.
  • the radical polymerization initiator is an alkyl halide and the control agent is a halogen atom involved in a reaction reaction.
  • the "active ingredient initiator” method implies a controlled radical polymerization technique using a control agent modified by chemical coupling with an active ingredient.
  • the modified control agent carries a molecule of active principle and the polymer obtained also carries an active ingredient per polymer chain at the proximal end.
  • alkyl any saturated linear or branched hydrocarbon chain, from 1 to 20 carbon atoms, preferably from 1 to 6 carbon atoms, such as for example methyl, ethyl, n-propyl, isopropyl, n-butyl sec-butyl, isobutyl, tert-butyl, pentyl and isomers thereof (eg n-pentyl, iso-pentyl), hexyl and isomers thereof (eg, n-hexyl, isohexyl).
  • arylalkyl refers to an alkyl group substituted with an aryl group and may be written as: aryl-alkyl-.
  • aryl refers to a polyunsaturated aromatic hydrocarbyl group having a single ring (e.g., phenyl) or multiple fused (e.g., naphthyl) or covalently linked aromatic rings typically containing from 5 to 12 carbon atoms; preferably 6 to 10, wherein at least one ring is aromatic.
  • the aromatic ring may optionally include one to two additional rings (either cycloalkyl, heterocyclyl or heteroaryl) fused thereto.
  • Non-limiting examples of aryl groups include phenyl, biphenylyl, biphenylenyl, 5 or 6 tetralinyl, naphthalen-1- or -2-yl, 4, 5, 6 or 7-indenyl, 1- 2-, 3-, 4 or 5-acenaphthylenyl, 3-, 4- or 5-acenaphthenyl, 1- or 2-pentalenyl, 4- or 5-indanyl, 5-, 6-, 7- or 8-tetrahydronaphthyl, 1, 2,3,4 tetrahydronaphthyl, 1,4-dihydronaphthyl, 1-, 2-, 3-, 4- or 5-pyrenyl.
  • “Pharmaceutically acceptable excipient” refers to an inert vehicle or carrier used as a solvent or diluent in which the pharmaceutically active agent is formulated and / or administered, and which does not produce an adverse, allergic or other reaction when administered to an animal, preferably a human being. This includes all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, absorption retardants, surfactants such as surfactant polymers, lipids and other similar ingredients. The selection of pharmaceutically acceptable excipients may be made by those skilled in the art depending on the properties of the nature and properties of the pharmaceutically active agent, the subject to be treated, and the route of administration. For human administration, preparations must respond to standards of sterility, general safety and purity, as required by regulatory authorities, such as the FDA or GEMA, for example.
  • “Pharmaceutically or therapeutically effective amount” means the necessary and sufficient amount of a pharmaceutical or therapeutic agent to be administered to a subject for slowing or stopping progression, aggravation, deterioration of at least one of the symptoms of 'a sickness. This amount administered can allow the relief of the symptoms of a disease or the cure of this disease.
  • “Pharmaceutical” refers to a compound or active principle in the field of health, having for example therapeutic properties and / or useful for a therapeutic diagnosis, especially for the purposes of treatment (curative and / or symptomatic and / or prophylactic) d 'a disease or therapeutic research.
  • the pharmaceutical term therefore includes therapeutic molecules or active principles and those of diagnosis.
  • Aqueous medium relates to a medium based on water molecules (H 2 0), in particular an aqueous solution.
  • an aqueous medium comprises between 50% and 100% water, by weight relative to the total mass of the medium.
  • An aqueous medium may especially be a biological fluid such as blood, lymph, saliva or urine.
  • active molecule and “active principle” are synonymous and relate to a compound for therapeutic use relating to health.
  • an active molecule may be indicated for treating or preventing a disease, preferably in a subject. This is called a pharmaceutically active molecule.
  • treatment of a disease means the reduction or amelioration of at least one undesirable effect or symptom of a disease, disorder or condition associated with a disability. of a function of an organ, a tissue or a cell.
  • preventing a disease or “inhibiting the development of a disease” relates to preventing or preventing the appearance of symptoms.
  • active ingredient is meant a compound having in particular therapeutic properties and / or useful for a therapeutic diagnosis, especially for the treatment (curative and / or symptomatic and / or prophylactic) of a disease or therapeutic research.
  • Prodrug refers to pharmacologically acceptable derivatives of the active molecule compounds, such as, for example, amides or esters, whose in vivo biotransformation product generates the biologically active molecule.
  • Prodrugs are generally characterized by an increase in bioavailability and are readily metabolized to biologically active compounds in vivo.
  • a prodrug is a water-soluble polymer covalently bonded to an active molecule.
  • Polymeric Prodrug This term refers to the polymer conjugated to at least one active ingredient.
  • Subject refers to an animal, including a human being.
  • a subject may be a patient, namely a person receiving medical care, undergoing or having undergone medical treatment, or monitored as part of the development of a disease.
  • the subject is treated for the first time.
  • the subject is resistant to another type of treatment and is treated with the prodrugs of the present invention as part of a second, third or fourth intention.
  • Upper critical temperature of solubility "Upper Critical Solution Temperature” and “UCST” are synonymous and relate to the critical temperature above which a thermosensitive polymer is completely soluble.
  • Thermosensitive relates to a property of a polymer whose physical properties change abruptly as a function of temperature.
  • the property concerned is the solubility of the polymer in an aqueous medium.
  • a heat-sensitive polymer has a higher critical solubility temperature (UCST).
  • the polymeric prodrugs according to the invention are obtained by polymerization of a monomer or comonomers.
  • the polymeric prodrugs according to the invention are obtained by controlled radical polymerization.
  • the polymeric prodrug of the invention comprises an active molecule covalently linked to a hydrophilic polymer chain by the initiator active principle method.
  • the PA is coupled to a polymerization control agent before the polymerization according to the so-called initiator active principle method.
  • PA is covalently coupled to a polymerization control agent. Once this coupling has been carried out, it is possible to grow a vinyl polymer in a controlled manner from this adduct control agent / PA. The PA will eventually end up covalently coupled to the proximal end of the polymer.
  • polymeric prodrugs Unlike other methods of synthesis of polymeric prodrugs (in particular that consisting of a coupling between the PA and a preformed polymer, called post-functionalization or that coupling PA to the monomer before polymerization), this technique makes it possible to position an AP at an end of each polymer chain.
  • the resulting polymeric prodrugs have a well-defined structure, a high loading rate and a simple purification. It is easily transposable to a large number of PAs and polymers.
  • the polymeric prodrug of the invention comprises an active molecule covalently linked to a water-soluble polymer chain by the initiator active principle method.
  • the controlled radical polymerization of the acrylamide monomers is carried out in the presence of the first molecule coupled to the chain transfer agent to form the polymer prodrug.
  • the polymeric prodrug is composed of polyacrylamide, of hydrophilic polyacrylamide derivatives, or of a polyacrylamide copolymer such as poly (acrylamide-co-acrylonitrile).
  • the polymer according to the invention comprises a polyacrylamide structure whose repeat unit has the formula [-CH 2 -CH (CONH 2 ) -] ", where n represents the number of repeating units in the polymer (or copolymer).
  • acrylamide monomer derivatives such as N-hydroxyacrylamide, N- (4-hydroxybutyl) methacrylamide, N- (poly (ethylene glycol)) -acrylamide, N- (3-methoxypropyl) methacrylamide, N - (2- (dimethylamino) ethyl) -N-methylmethacrylamide, N- (2- (diethylamino) ethyl) -N-methylmethacrylamide.
  • the method or method according to the invention advantageously makes it possible to provide a polymer prodrug having a pharmaceutically active molecule (or PA) at one end of the prodrug-polymer molecule and thus advantageously makes it possible to control the loading rate of the AP.
  • a control is not present in the prior art in which the AP is coupled either after polymerization or to the monomer before polymerization.
  • a second pharmaceutically active molecule is coupled by covalent coupling to the terminal portion of the control agent. This coupling takes place after the end of the radical polymerization.
  • the choice of the polymer and its size makes it possible to obtain polymeric prodrugs that can be administered SC or IM.
  • the size of the polymeric prodrugs is controlled by the radical polymerization conditions.
  • the polymeric prodrug comprises an active molecule covalently linked to a polyacrylamide chain by the initiator active ingredient method.
  • the polymer prodrug comprises an active molecule covalently bound to a copolymer by the initiator active ingredient method obtained from acrylamide monomers and one or more other comonomers.
  • the polymeric prodrug comprises an active molecule covalently linked with a copolymer by the initiator active ingredient method obtained from acrylamide and acrylonitrile monomers to provide the UCST poly (acrylamide) thermosensitive polymer prodrug. r -acrylonitrile),
  • the polymeric prodrug comprises an active molecule covalently bound to a polymer by the initiator or copolymer active principle method obtained from hydrophilic monomers derived from acrylamide such as N-hydroxyacrylamide, N- ( 4-hydroxybutyl) methacrylamide, N- (poly (ethylene glycol)) -acryalamide, N- (3-methoxypropyl) methacrylamide, N- (2- (dimethylamino) ethyl) -N-methylmethacrylamide, N- (2- (diethylamino) ethyl) -N-methylmethacrylamide.
  • acrylamide such as N-hydroxyacrylamide, N- ( 4-hydroxybutyl) methacrylamide, N- (poly (ethylene glycol)) -acryalamide, N- (3-methoxypropyl) methacrylamide, N- (2- (dimethylamino) ethyl) -N-methylmethacrylamide, N- (2- (die
  • the copolymer according to the invention is a random copolymer.
  • the copolymer according to the invention is a random copolymer.
  • the polymer of the polymeric prodrug according to the invention is not crosslinked.
  • the polymeric prodrug according to the invention does not form a crosslinked hydrogel.
  • a molar mass that is significantly greater than that of the active molecule improves the maintenance of the properties related to the interactions between the polymer and the solvent, and in particular improves the solubility in the aqueous phase of the active molecule.
  • the polymer of the invention has a molecular weight of 1,000 to 100,000 g / mol.
  • the polymer of the invention has a molecular weight of 2,000 to 70,000 g / mol, 5,000 to 70,000 g / mol, 5,000 to 60,000 g / mol, 5,000 to 50,000 g / mol, 5,000 to 40,000 g / mol, from 5,000 to 30,000 g / mol, from 5,000 to 40,000 g / mol, from 10,000 to 40,000 g / mol, from 15,000 to 40,000 g / mol, from 15,000 to 30,000 g / mol or from 20,000 to 30,000 g / mol.
  • the molar mass of the polymer is 1,000 to 80,000 g / mol.
  • the terminal portion of the polymer prodrug varies the solubility and / or the viscosity of the polymeric prodrug.
  • the polymer chain for example of polyacrylamide, comprises an alkyl terminal chain, for example comprising from 2 to 20 carbon atoms or a -SH, -COOH, -NH 2 , halogen function.
  • the length of the alkyl chain or its nature makes it possible to vary the viscosity of the polymeric prodrug.
  • the polymeric prodrug is thermally sensitive, having a higher critical solubility temperature (UCST) of 0 to 60 ° C in an aqueous medium.
  • the polymeric prodrug comprises an active molecule that is covalently bonded to a thermosensitive polymer chain of poly (acrylamide-co-acrylonitrile).
  • the poly (acrylamide-co-acrylonitrile) chain of these polymeric prodrugs has a molar mass of 1,000 to 100,000 g / mol.
  • the molar percentage of acrylonitrile is more than 0 to 100% preferably from 1 to 50%, and more preferably from 5 to 35% relative to the number of moles of the polymer.
  • the pharmaceutically active molecule to be used for the preparation of the polymeric prodrugs of the invention is a free molecule or a molecule bound with another molecule.
  • the active molecule for the preparation of the polymeric prodrugs of the invention is free.
  • the pharmaceutically active molecule to be used for the preparation of the polymeric prodrugs of the invention has a function capable of reacting with a controlled radical polymerization agent according to the invention so as to covalently couple the active molecule (or AP). and the polymerization agent.
  • the free function is a nucleophilic function.
  • the pharmaceutically active molecule may not carry free functions. It can be chemically treated prior to its coupling to the agent of polymerization so that it has a free function (capable of reacting with the controlled radical polymerization agent).
  • Several approaches are known in the art for functionalizing an active molecule.
  • An illustrative and non-limiting example for the present invention is the treatment of an active molecule with hyperoxides leading to hydroxylation of the active molecule.
  • the free nucleophilic function of the active molecule is selected from the groups -OH, -NH 2 , -NHR and -SH.
  • the nucleophilic function is -OH or -NH 2 .
  • the other nucleophilic functions of the molecule if it has any, can be protected or not.
  • the nucleophilic functions of the active molecule that do not participate in the controlled radical polymerization are not protected.
  • the nucleophilic functions of the active molecule which do not participate in the controlled radical polymerization are protected by groups known in the art such as tert-butoxycarbonyl chloride, d-butylcarbyl dicarbonate, azide or amide of / e / 7-butoxycarbonyl, tert-butyl (dimethyl) silyl chloride, tosyl chloride, alkyls, aryls, alkylaryls, esters, ethers, silyl ethers,
  • groups known in the art such as tert-butoxycarbonyl chloride, d-butylcarbyl dicarbonate, azide or amide of / e / 7-butoxycarbonyl, tert-butyl (dimethyl) silyl chloride, tosyl chloride, alkyls, aryls, alkylaryls, esters, ethers, silyl ethers,
  • the invention can be implemented independently of the polarity of the active molecule. Therefore, the active molecule for use in the polymers of the invention is a polar, amphiphilic or apolar molecule. According to a first embodiment, the active molecule to be used in the polymers of the invention is a polar molecule. According to a second embodiment, the active molecule to be used in the polymers of the invention is an apolar molecule. According to a third embodiment, the active molecule to be used in the polymers of the invention is an amphiphilic molecule.
  • antibiotic molecules bacteri / fungo-static, bacterio / fungo-toxic or anti viral agents
  • the active molecule is an anticancer molecule selected from: paclitaxel, docetaxel, gemcitabine, cladribine, capecitabine, daunorubicin, doxorubicin, epirubicin, ridarubicin, ractinomycin, ramsacrine, dacarbazine, dactinomycin, vincristine, vimblastine, vindesine, methotrexate, colchicine, cyclophosphamide, azathioprine, 6-mercaptopurine, lomustine, carmustine, dacarbazine, cisplatin, fluorouracil, tenoposide or etoposide, fotemustine, mitomycin C, mitoxantrone, streptozocin, trabectedin, vinflunine, vinorelbine, asernic trioxide, bendamustine, busulfan, cabazitaxel, carboplatin, erib
  • the active molecule is chosen from anticancer molecules (paclitaxel, gemcitabine), from peptides (cyclic RGD) and or fluorescent probes (rhodamine and Cyanine 5.5).
  • the active molecule is an anticancer molecule selected from paclitaxel or gemcitabine.
  • the prodrugs of the invention comprise paclitaxel as the active molecule.
  • the first active molecule is paclitaxel.
  • the first active molecule is gemcitabine.
  • a polymeric prodrug according to the invention comprises a molecule active at one end.
  • a polymeric prodrug according to the invention comprises an active molecule at its proximal end.
  • a polymeric prodrug according to the invention comprises an active molecule at its terminal end.
  • a polymeric prodrug according to the invention comprises two active molecules, one at each end.
  • the present invention relates to molecules on which a polymer chain, in particular as described above, is grafted.
  • the polymerization according to the invention is carried out by a controlled radical route.
  • the polymerization according to the invention is carried out by a radical route controlled by the initiator active principle method.
  • the polymer chain is grafted onto the active molecule by applying a controlled radical polymerization process.
  • the polymer prodrug of the invention is obtained by a controlled radical polymerization process chosen from:
  • RAFT Reversible Addition-Fragmentation Chain Transfer
  • NMP nitroxide-controlled radical polymerization
  • ATRP controlled radical polymerization Atom Transfer Radical Polymerization
  • SET-LRP Single Electron Transfer- Living Radical Polymerization
  • the polymeric prodrug further comprises a chain transfer agent.
  • the polymeric prodrug is synthesized by reversible addition-fragmentation transfer controlled radical polymerization (hereinafter "RAFT").
  • RAFT reversible addition-fragmentation transfer controlled radical polymerization
  • the polymeric prodrugs according to the invention are obtained by controlled radical polymerization of controlled radical polymerization type controlled by reversible transfer by addition-fragmentation (hereinafter "RAFT"), by reacting at least one monomer, an initiator of radical polymerization and a controlled radical polymerization control agent (also called chain transfer agent) on which is coupled the pharmaceutically active molecule.
  • RAFT controlled radical polymerization type controlled by reversible transfer by addition-fragmentation
  • a polymeric prodrug according to the invention is prepared by controlled radical polymerization and comprises, according to one variant, a covalent coupling of at least one first pharmaceutically active molecule with a radical polymerization control agent comprising a proximal portion and a terminal portion to form a first coupled molecule.
  • the radical polymerization control agent comprises a proximal portion and an end portion because during the polymerization the polymerization control agent (or chain transfer agent) is cleaved with a portion, here called proximal remaining connected to the PA which will be positioned at the beginning of the polymer chain and a part, here called terminal, which is fixed at the end of the polymer chain to complete.
  • This polymerization control agent makes it possible to precisely and advantageously control the length of the polymer chain.
  • first coupled molecule refers to PA coupled to the polymerization control agent or to the proximal portion of the control agent.
  • the polymer of the prodrug of the invention further comprises a RAFT chain transfer agent selected from:
  • Trithiocarbonates such as 3,5-bis (2-dodecylthiocarbonothioylthio-1-oxopropoxy) benzoic acid, 3-butenyl 2- (dodecylthiocarbonothioylthio) -2-methylpropionate, 2- (2-carboxyethylsulfanylthiocarbonylsulfanyl) propionic acid 4 - ((((2-carboxyethyl) thio) carbonothioylthio) -4-cyanopentanoic acid, 2-cyanobutan-2-yl 4-chloro-3,5-dimethyl-1H-pyrazole-1-carbodithioate 2-cyanobutanyl-2-yl 3,5-dimethyl-1H-pyrazole-1-carbodithioate, 4-cyano-4 - [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid, 2- (butylthiocarbon
  • Dithiocarbamates such as benzyl 1H-pyrrole-1-carbodithioate, cyanomethyl diphenylcarbamodithioate, cyanomethyl methyl (phenyl) carbamodithioate, cyanomethyl methyl (4-pyridyl) carbamodithioate, 2-cyanopropan-2-yl N-methyl-
  • N- (pyridin-4-yl) carbamodithioate methyl 2- [methyl (4-pyridinyl) carbamothioylthio] propionate, 1-succinimidyl-4-cyano-4- [N-methyl-N- (4-pyridyl) carbamothioylthio) pentanoate;
  • Dithioabenzoates such as benzyl benzodithioate, cyanomethyl benzodithioate, 4-cyano-4- (phenylcarbonothioylthio) pentanoic acid, 4-cyano-4- (phenylcarbonothioylthio) pentanoic acid N-succinimidyl ester, 2- cyano-2-propyl benzodithioate, 2-cyano-2-propyl 4-cyanobenzodithioate, G ethyl 2- (4-methoxyphenylcarbonothioylthio) acetate, G ethyl 2-methyl-2- (phenylthiocarbonylthio) propionate, G ethyl 2- (phenylcarbonothioylthio) Phenylacetate, G ethyl 2- (phenylcarbonothioylthio) propionate, 1-
  • Switchable RAFT agents such as cyanomethyl methyl (4-pyridyl) carbamodithioate, 2-cyanopropan-2-yl N-methyl-N- (pyridin-4-yl) carbamodithioate, methyl 2- [methyl (4-pyridinyl) carbamothioylthio] propionate or 1-succinimidyl-4-cyano-4- [N-methyl-N- (4-pyridyl) carbamothioylthio] pentanoate.
  • the chain transfer agent is for example chosen from: 4-cyano-4- [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid, 2- (dodecylthiocarbonothioylthio) -2-methylpropionic acid, 4-cyano-4- acid
  • the conventional radical initiator is chosen from: 2,2'-azobis (2-methylpropionitrile), 1,1'-azobis (cyclohexanecarbonitrile), 4,4'-azobis (4-cyanovaleric acid), 2 , 2'-azobis (2-methylbutyronitrile), benzoyl peroxide, lauroyl peroxide, methyl ethyl ketone peroxide, tert-butyl peroxybenzoate.
  • the chain transfer agent selected from: 4-cyano-4 - [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid, G acid 2-
  • the chain transfer agent is selected from:
  • the chain transfer agent is 4-cyano-4- [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid ("CDP" hereinafter).
  • the transfer agent is directly linked to the active molecule.
  • the transfer agent by virtue of its polymerization control function is split into two parts, one proximal which remains linked to the active ingredient and the other which reacts with the terminal part. growing polymer.
  • the transfer agent is covalently bound to the pharmaceutically active molecule by an ester, amide, carbonate, carbamate, acetal, disulfide, thioether or triazole function; diglycolate linker, succinate, succinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC), glycinate, glucuronate, valine-citrulline, maleimide, said function and / or proximal linker forming a covalent bond with the first pharmaceutically active active ingredient.
  • SMCC succinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate
  • glycinate glycinate
  • glucuronate glucuronate
  • valine-citrulline valine-citrulline
  • maleimide said function and / or proximal linker forming a covalent bond with the first pharmaceutically active active ingredient.
  • the transfer agent is previously functionalized with an oligomer of a hydroxy carboxylic acid.
  • this oligomeric chain which is located after grafting between the active molecule and the transfer agent, makes it possible to control the rate of release of the active molecule of the polymeric prodrug of the invention.
  • the oligomer G is a dimer, preferably diglycolic acid.
  • a radical polymerization initiator is necessary because it allows the initiation of the polymerization and thus to grow the polymer chain from the active molecule functionalized by the RAFT agent.
  • the initiator may be of azo type such as 2,2'-azobis (2-methylpropionitrile), 1,1'-azobis (cyclohexanecarbonitrile), 4,4'-azobis (4-cyanovaleric acid), 2,2'-azobis (2-methylbutyronitrile); inorganic peroxide type; or organic peroxide type such as benzoyl peroxide, lauroyl peroxide, methyl ethyl ketone peroxide, tert-butyl peroxybenzoate.
  • the initiator of free radicals is 2,2'-azobis (2-methylpropionitrile), CAS No. 78-67-1.
  • the polymeric prodrug of the invention is a polymeric prodrug whose active molecule is covalently linked with a block copolymer.
  • the latter comprises the polymer or copolymer as described above and an extension with at least one hydrophilic polymer. Indeed, the presence of the transfer agent at the end of the polymer or copolymer chain allows the addition of an additional polymer chain.
  • the copolymer of the invention may comprise at least two blocks.
  • the prodrug of the invention comprises a polymer whose chain further comprises an extension of its chain by an additional polymer, preferably the additional polymer being a water-soluble polymer.
  • the water-soluble polymer is chosen from poly [oligo (ethylene glycol) methyl ether methacrylate], poly [oligo (ethylene glycol) methyl ether acrylate] poly [oligo (ethylene glycol) methacrylate], polyacrylamide, glycopolymers (synthesized from monomers of methacrylate, acrylate, acrylamide vinyl ether or styrenic type carrying a sugar function), poly (/ V, / V-di methyl acrylamide), polystyrene sulfonate, poly (/ V-vinyl pyrrolidone), hydrophilic polypeptides and polysaccharides.
  • poly [oligo (ethylene glycol) methyl ether methacrylate] poly [oligo (ethylene glycol) methacrylate]
  • polyacrylamide glycopolymers (synthesized from monomers of methacrylate, acrylate, acrylamide vinyl ether or styrenic type carrying a sugar function)
  • the water-soluble polymer is polyacrylamide.
  • the invention relates to the process for preparing the polymer, as described above.
  • Process for obtaining the polymer is described above.
  • the method comprises at least one polymerization step from the active molecule.
  • the process for preparing a polymer according to the invention comprises the steps of:
  • the method comprises at least one controlled radical polymerization step from the active molecule.
  • the controlled radical polymerization may be selected from the techniques known in the art such as:
  • RAFT reversible addition-fragmentation Chain Transfer
  • NMP nitroxide-controlled radical polymerization
  • ATRP atom transfer controlled radical polymerization
  • the process for preparing the polymeric prodrug of the invention comprises at least one RAFT polymerization step.
  • the process for preparing a polymer according to the invention comprises the steps of:
  • step (ii) is carried out in the presence of a radical polymerization initiator (free radical generator).
  • the polymerization initiator allows the initiation of the polymerization and thus to grow the polymer chain from the active molecule functionalized by the RAFT agent.
  • the initiator may be of the azo type such as 2,2'-azobis (2-methylpropionitrile), 1,4-azobis (cyclohexanecarbonitrile), 4,4'-azobis (4-cyanovaleric acid), 2 2'-azobis (2-methylbutyronitrile); inorganic peroxide type; or organic peroxide type such as benzoyl peroxide, lauroyl peroxide, methyl ethyl ketone peroxide, tert-butyl peroxybenzoate.
  • the free radical initiator is 2,2'-azobis (2-methylpropionitrile), CAS No. 78-67-1.
  • step (ii) is performed at room temperature. In another embodiment, step (ii) is carried out at a temperature of 25 ° to 150 ° C.
  • the present invention has the advantage of providing a platform that can be applied to a wide variety of active ingredients, imaging agents and / or targeting agents.
  • a functional group at each end of a polymer chain can be used to chemically link the molecule of interest thus making it possible to manufacture mono- or bifunctional systems called telechelic polymers. It is possible to design a large number of systems that range from the simplest system (a molecule grafted at one end of the polymer chain) to more complex so-called heterobifunctional systems where one can have a pharmaceutically active molecule at a higher level. end and an imaging agent or targeting ligand at the other end of the polymer.
  • various polymeric prodrugs with different active ingredients can also be synthesized and then formulated together to produce combination therapies.
  • This type of telechelic polymers of macromolecular architecture is particularly interesting because it will be possible to create polymeric prodrugs with different properties and applicable to many fields.
  • the heterobifunctional polymers are more particularly interesting because they make it possible to combine two different functionalities in the same compound.
  • These polymeric prodrugs according to the invention are therefore particularly suitable for biomedical applications, where it would be possible to conjugate on the same chain two types of molecules that can be pharmaceutically / biologically active for different purposes, for example:
  • a targeting ligand e.g., antibody, ligand, peptide, etc.
  • a targeting ligand for imaging, by combining a targeting ligand and a tracing molecule such as a fluorophore.
  • a tracing molecule such as a fluorophore.
  • the method for preparing telechelic polymer prodrugs comprises one or more pre- or post-polymerization modifications for coupling the different molecules of interest.
  • a polymeric prodrug comprising a first active ingredient to a peptide, typically a peptide coupling, is coupled by covalent bonding.
  • the first route makes it possible to associate the peptide with the polymer via a maleimide-type linker by a thiol-maleimide coupling
  • the second route makes it possible to carry out the coupling directly by forming a peptide bond. by peptide coupling.
  • the polymerization is initiated by the peptide and then the coupling with the active principle is carried out after polymerization.
  • the invention relates to a composition comprising at least one polymeric prodrug of the invention.
  • compositions comprise a single polymeric prodrug of the invention.
  • the compositions comprise at least two, at least three, at least four or at least five polymeric prodrugs of the invention.
  • the compositions comprise polymeric prodrugs comprising the same active molecule.
  • the presence of polymeric prodrugs with different polymer chains allows a bimodal, tri-modal or multi-modal release of the active molecule.
  • the compositions comprise polymeric prodrugs comprising different active molecules and the same polymer chain. This makes it possible to have in the same formulation two active molecules with a different pharmacodynamic profile.
  • the compositions comprise polymeric prodrugs comprising active molecules and different polymer chains. This allows to have in the same formulation two active molecules with a different pharmacodynamic profile whose release is adapted according to these physicochemical properties.
  • compositions comprise at least one polymeric prodrug of the invention and at least one free active molecule, its pharmaceutically acceptable salts or its prodrugs as known in the art.
  • free active molecule is meant a molecule not bound, or at least not covalently bonded with the polymer.
  • the present invention relates to compositions in the form of an aqueous solution comprising an aqueous medium and at least one polymeric prodrug according to the invention.
  • the polymer prodrug according to the invention is soluble in an aqueous medium.
  • the polymeric prodrug according to the invention is soluble in distilled water at least 100 mg / mL and preferably 150 mg / mL and more preferably at 200 mg / mL.
  • the solubility of the polymeric prodrug is preferably tested according to the following method:
  • the different polymeric prodrugs are dissolved in an equivalent concentration (200 mg / ml) and then centrifuged for 30 min at 16,783 g. Non-solubility is observed by the appearance of a whitish or colored deposit in the bottom of the Eppendorf.
  • the solution comprising the polymeric prodrug according to the invention is not very viscous.
  • the viscosity of the solution of the polymer prodrug can be modulated according to the size, nature / composition of the polymer.
  • the controlled radical polymerization here again has an important technical advantage for the invention.
  • the polymeric prodrug according to the invention makes it possible to concentrate the AP without significantly increasing the viscosity of the formulation, which makes it possible to limit the quantities (in particular by volume) of the injected formulation.
  • a polymeric prodrug according to the invention is formulated in injectable form.
  • the polymer prodrug according to the invention is formulated in aqueous solution easily injectable.
  • the viscosity of the solution comprising a polymeric prodrug according to the invention allows it to be injected via a 26 g syringe.
  • the solution comprising a polymeric prodrug according to the invention requires an injection force through a needle. of 26 G of less than 30 N.
  • the solution comprising a polymeric prodrug according to the invention is injectable through a 26 G needle at a concentration of at least 50 mg / mL, for example from less 100 mg / mL and preferably at least 125 mg / mL.
  • the solution comprising a polymeric prodrug according to the invention is injectable through a 26 G needle at a concentration of at least 150 mg / ml and preferably at least 200 mg / ml.
  • injectability (or syringability) (expressed in newton as a function of concentration) is tested according to the following method:
  • the syringability / injectability of the polymer solutions is estimated by custom-made equipment as described by Burckbuchler et al. (Eur J Pharm Biopharm, 76, 2010, 351-356) coupled to a texture analyzer (TA.XT Plus Texture Analyzer, Stable Micro Systems) having a force sensor of 30 kg. 400 ⁇ l of each solution are taken and then injected through a 1 ml syringe (MeritMedical, Médaillon® Syringe) and a 26G x 1 ⁇ 2 "needle (Terumo Neolus, 0.45x12 mm) at a speed of 1 mm / s. The injection force is measured at 25 measurements per second.
  • TA.XT Plus Texture Analyzer Stable Micro Systems
  • the polymer makes it possible to increase the solubility of PA, to maintain stability at high concentration, to maximize its absorption and to limit its metabolism, thus leading to increased bioavailability.
  • the present invention relates to a polymeric prodrug for use in a method of therapeutic treatment.
  • the method comprises administering a therapeutically effective amount of the polymeric prodrug to a patient.
  • the prodrug approach (where the AP is inactive until its release) makes it possible to eliminate the local undesirable effects of the necrotizing / irritant APs.
  • the active ingredient is released from the polymer by cleavage of the bond and then regains its activity. Cleavage is obtained because of the biological conditions present in the bloodstream.
  • the active ingredient is released from the prodrug by cleavage of the link and then finds its activity.
  • the polymeric prodrug is injected into a tissue (SC or IM in particular) and passes into the bloodstream. The polymeric prodrug is then cleaved to release the PA of interest into the bloodstream.
  • the control of the nature of the monomer, the size and the dispersity of the polymer, the nature of the polymerization control agent and the bond between it and the PA makes it possible to modify the rate and the rate.
  • polymeric prodrugs of the invention as well as their compositions may have several applications in the biomedical field.
  • the AP designates an active molecule linked to a targeting agent making it possible to target a specific area to be treated and for example to direct the released active molecule towards its site of action.
  • the AP designates an active molecule linked to a diagnostic agent that makes it possible to image a specific area to be treated.
  • the AP designates a targeting agent linked to a diagnostic agent making it possible to target the AP towards the tissue or cells to be treated.
  • the invention also relates to a medicament comprising at least one polymeric prodrug of the invention.
  • the invention further relates to the use of the polymeric prodrug according to the invention for the prevention and / or treatment of a disease, in particular a human or animal.
  • the invention also relates to the use of at least one prodrug according to the invention for the preparation of a medicament.
  • the drug comprises at least one polymeric prodrug of the invention in a therapeutically effective amount.
  • the medicament further comprises pharmaceutically acceptable excipients. These excipients correspond to the standards of the European Pharmacopoeia or the FDA.
  • a drug formulation is determined by the skilled person according to the disease to be prevented and / or treated, the route of administration of the drug and the nature of the active molecule.
  • the formulations are injectable formulations.
  • the administration is by bolus or continuous (infusion), preferably the administration is by bolus.
  • the formulations are injectable formulations with administration:
  • a polymeric prodrug according to the invention is administered (or administrable) subcutaneously or intramuscularly.
  • the invention also relates to a polymeric prodrug for use in a method of therapeutic treatment.
  • This method comprises administering a therapeutically effective amount of at least one polymeric prodrug of the invention to a subject, particularly a human or animal.
  • the methods of treatment may relate to the treatment of diseases as described above.
  • the method of treatment is a method of treating cancer.
  • the administration of at least one polymeric prodrug according to the invention may be simultaneous with the administration of other active molecules, formulated according to the invention or not.
  • the administration of at least one polymer prodrug according to the invention may be sequential to the administration of other active molecules, formulated according to the invention or not.
  • these formulations make it possible to increase the bioavailability of the active principle and are administered subcutaneously, intramuscularly, intratumorally or intradermally, preferably subcutaneously or intramuscularly.
  • the diseases that can be prevented or treated by the medicament of the invention are nonlimitingly chosen from cancers, bacterial infections, viral infections, fungal infections, parasitic infections, inflammatory diseases, metabolic diseases, microvascular diseases, macro-vascular diseases, cardiovascular diseases, pulmonary diseases, endocrine diseases or diseases of the central nervous system .
  • the type of cancers that can be treated by the administration of a polymeric prodrug according to the invention are not particularly limited since the treatment depends on the grafted active molecule.
  • the active molecule grafted onto the polymer is chosen according to its biological, pharmacodynamic and pharmacokinetic properties in relation to the cancer to be treated.
  • the treated cancer is a solid cancer such as breast cancer, liver cancer, melanoma, ovarian or endometrial cancer, prostate cancer and / or bladder cancer, stomach, bowel, Kaposi's sarcoma, brain cancer, bone cancer, pancreatic cancer or lung cancer.
  • the cancer is a cancer of blood cells such as leukemia.
  • the active molecule is released rapidly, for example 80% by weight of the active molecule is released in less than 24 hours. According to one embodiment, the active molecule is released slowly, for example 50% by weight of the active molecule is released in addition 72h.
  • FIG. 1 shows three Transmittance spectra as a function of the temperature of 3 copolymers according to the invention having different RAFT agents: CDP-33% -5, PTX-CDP-33% -10 and Gem-33 -5%. Acquisition at 4.5 mg / ml in PBS at 600 nm and at a temperature increase rate of 0.5 ° C / min.
  • FIG. 2 At the top, shows Transmittance vs. curves. Temperature showing the UCST behavior of the CP5-PEGMA and CP7-PEGMA particles at a polymer concentration of 4.5 mg / mL. Bottom, shows the comparative curve UCST behaviors of CP5 and CP5-PEGMA obtained after PEGylation and formulation of CP5.
  • FIG. 3 Top (A), shows the local toxicity effects of subcutaneous administration of a solution of paclitaxel in PBS.
  • Bottom (B) shows the absence of local toxicity following the subcutaneous administration of a paclitaxel solution formulated in prodrug according to the invention PTX-P (AAm-co-AN) with 20% of AN .
  • Figures 4 and 5 Graphs of the results of the viscosity studies of different polymeric prodrugs by seringability (Force (N) versus concentration (mg / mL)).
  • Figures 6 and 7 release in PBS ( Figure 6) and in the murine plasma ( Figure 7) paclitaxel from the prodrug polymers having different chemical bonds between the PTX and the polymer (ester, diglycolate, carbonate).
  • Figure 8 Pharmacokinetics in a mouse model of PTX released into the plasma after degradation of the polymer binding (concentration (mg / mL) versus time in minutes) determined by mass spectrometry.
  • Figure 9 Toxicity study in a murine model. Variation in mouse weight as a function of time for PTX-digly-PAAm, PTX-Ester-PAAm, Taxol and PAAm.
  • Figure 10 Pharmacokinetics and biodistribution of radio-labeled PTX in the commercial formulation of Taxol® or in the form of PTX-PAAm administered at 7 mg / kg equivalent of PTX (0.14 mg total PTX per mouse) intravenously and under -cuta Amsterdam. Results for PTX-PAAm allow for free PTX plus PTX coupled with PAAm.
  • Figure 11 Biodistribution study in a mouse model of free rhodamine and Rhodamine-PAAm administered IV and SC.
  • Figure 12 Top, shows the effects of local toxicity of subcutaneous administration of paclitaxel solution in PBS. Bottom, shows the absence of local toxicity following the subcutaneous administration of a solution of paclitaxel formulated in prodrug according to the invention PTX-PAAm.
  • Figure 13 Viscosity of solutions of prodrug paclitaxel polymer as a function of the concentration and the content of acrylonitrile (AN) in the polymer. These measurements were obtained using a rheometer with plane-plane geometry.
  • AAm refers to acrylamide - CAS No. 79-06-1
  • AIBN designates the radical initiator Azobisisobutyronitrile - CAS n ° 78-67-1
  • AN refers to acrylonitrile - CAS No. 107-13-1
  • CDP refers to the RAFT control agent 4-cyano-4- [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid - CAS No. 870196-80-8
  • CDP-ol refers to the functionalizing agent RAFT 4-cyano-4- [(dodecylsulfanylthiocarbonyl) sulfanyl] pentanol - CAS No. 1394136-26-5
  • Flash chromatography refers to a method of preparative separation.
  • the mobile phase passes through the stationary phase by applying a pressure of 10 to 20 psi
  • DCM refers to anhydrous dichloromethane - CAS No. 75-09-2
  • DMAP refers to 4-Dimethylaminopyridine - CAS No. 1122-58-3
  • DMF refers to N, N-Dimethylformamide - CAS No. 200-679-5
  • DMSO refers to Dimethylsulfoxide - CAS # 67-68-5
  • EDC-HC1 refers to 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride - CAS No. 25952-53-8
  • EtOAC refers to ethyl acetate - CAS No. 7487-88-9
  • Gem refers to Gemcitabine hydrochloride - CAS # 122111-
  • GemTBDMS refers to Gemcitabine whose alcohol functions are protected by tert-Butyldimethylsilyl
  • NHS N-hydroxysuccinimide - CAS No. 6066-82-6
  • PEGMA Poly (ethylene glycol) methyl ether methacrylate, average Mn 300 g / mol - CAS No. 26915-72-0
  • PTX-A% -B denotes the paclitaxel-CDP-Poly polymer (AAm-co-AN) with a molecular weight of B kg / mol and comprising A% acrylonitrile relative to the moles of the polymer.
  • PTX-A% -B-PEGMA-C refers to the paclitaxel-CDP-Poly polymer (AAm-co-AN) with a molecular weight of B kg / mol and comprising A% acrylonitrile and C% PEGMA relative to moles of the polymer
  • PTX refers to paclitaxel - CAS No. 33069-62-4
  • Brine means a saturated aqueous solution of sodium chloride
  • TB AF refers to tetra-n-butylammonium fluoride - CAS No 429-4l-4
  • TBDMS refers to the tert-Butyldimethylsilyl radical
  • TBDMSC1 refers to tert-Butyldimethylsilyl chloride - CAS hr 18162-48-6
  • THF refers to tetrahydrofuran - CAS No. 109-99-9
  • UV-Visible Spectroscopy For transmittance measurements of aqueous polymer solutions at a fixed concentration of 4.5 mg / mL, the absorption spectra were recorded on a Perkin-Elmer Lambda 25 UV-Visible Spectrophotometer using a ramp rise and fall of temperature from 20 to 60 ° C at a rate of 0.5 ° C / min thanks to a Peltier effect system.
  • Viscosity is measured using seringability studies as a function of concentration.
  • the serability of the polymer solutions was estimated by custom-made equipment (Burckbuchler et al., Eur J Pharm Biopharm, 76, 2010, 351-356) coupled to a texture analyzer (TA.XT Plus Texture Analyzer, Stable Micro Systems) with a force sensor of 30 kg. 400 ⁇ l of each solution are taken and then injected through a 1 ml syringe (MeritMedical, Médaillon® Syringe) and a 26G x 1 ⁇ 2 "needle (Terumo Neolus, 0.45x12 mm) at a speed of 1 mm / s. The injection force is measured at 25 measurements per second. Each sample is injected 3 times in a row. By this method it is considered that a solution is difficult to inject if the force required for the injection exceeds 30 N.
  • Example 1 Coupling of paclitaxel (PTX) to the control agent 4-cyano-4-G (dodecylsulfanylthiocarbonylisulfanyllpentanoic (CPP)
  • PTX paclitaxel
  • 4-cyano-4-G dodecylsulfanylthiocarbonylisulfanyllpentanoic
  • This species corresponds to paclitaxel covalently coupled to the CDP control agent named in the following PTX-CDP examples.
  • This example shows the production of a paclitaxel-polyacrylamide polymer prodrug by RAFT type polymerization of acrylamide in the presence of PTX-CDP, the following image corresponds to the chemical structure of the synthesized polymer prodrug:
  • the polymers obtained have the following characteristics: PTX-PAAm M n 21,600, MJM at 1,12 (Example 2); PTX-PDMAAm M n 20,200, MJM n 1.02; PTX-POEGMA M n 24,500; PTX-PGMA M n 20,500, M w / M n 1.11; PTX-PEG M n 21,000, MJM n 1.05.
  • solubility is evaluated at a concentration of 200 mg / ml of polymeric prodrugs, to distinguish the soluble polymeric prodrugs from the polymer prodrugs suspended one or two centrifugations (16 873 g, 30 min) are carried out. Solubility is evaluated by the absence of visible aggregates at the Eppendorf pellet.
  • PAAm therefore allows complete solubilization of paclitaxel at 200 mg / ml.
  • the other prodrugs therefore have a lower solubility than PTX-PAAm.
  • Viscosity studies of different polymeric prodrugs by syringes Viscosity is a reflection of the solubility of a polymer. The phenomena of entanglement of the polymer chains are greater when the polymer prodrug is very soluble, which leads to an increased viscosity. Viscosity is measured through seringability studies as a function of concentration. The serability of the polymer solutions was estimated by custom-made equipment (Burckbuchler et al., Eur J Pharm Biopharm, 76, 2010, 351-356) coupled to a texture analyzer (TA.XT Plus Texture Analyzer, Stable Micro Systems) with a force sensor of 30 kg.
  • TA.XT Plus Texture Analyzer Stable Micro Systems
  • PTX-PAAm The viscosity of PTX-PAAm is higher than that of other polymers which is related to the higher entanglement of the polymer chains and therefore to the higher viscosity.
  • PAAm is the polymer with the highest capacity to solubilize highly hydrophobic PAs such as PTX.
  • Example 2 Then these were used to polymerize acrylamide as described in Example 2 to obtain the prodrugs PTX-PAAm-Cl2 (Example 2), PTX-PAAm-C4 and PTX-PAAm-C2 respectively.
  • the following diagram summarizes these different syntheses:
  • the polymers obtained have the following characteristics: PTX-PAAm-Cl 2 M n 21,600, MJM n 1,12 (Example 1); PTX-PAAm-C4 M n 26,400, M w / M n 1.04; PTX- PAAm-C2 M n 24,100, M w / M n 1.09.
  • the viscosity of the various prodrugs was measured by the same method described in Example 3, the results are grouped together in FIG. 5.
  • IR ⁇ 3270, 2924, 2854, 1635, 1546, 1439, 1379, 1200, 1182, 1075, 1130, 799, 720, 698, 606, 517 cm -1 .
  • Rhodamine Fluorescent Polymers The rhodamine (Rho) piperazine used was synthesized following a synthetic route already described by Nguyen et al. (Organic Letters, 2003, 5, 3245-3248). This is coupled to the RAFT CDP agent to obtain the Rho-CDP whose structure is given in the following figure:
  • This synthesis is carried out in two stages; a first is to react the CDP with glycolic anhydride in the presence of a base (triethylamine). The reaction is carried out at room temperature for 24 hours and quantitatively forms CDP-glycolic acid 1. A coupling between the latter with paclitaxel in the presence of EDC as coupling agent and DMAP as a base for 24 hours at 30 ° C. C gives the product PTX-digly-CDP 2 with a yield of 50%.
  • a base triethylamine
  • the first part consists of the formation of CDP-NH 2 .
  • the latter has never been described in the literature.
  • the CDP is activated in the presence of NHS and DCC to provide the product 4 with 60% yield.
  • the next step is to couple a diaminoethane chain with CDP-NHS to form a peptide bond.
  • diaminoethane is protected by a group (BOC).
  • Diaminoethane-BOC was added to a solutionCDCD-NHS in anhydrous DCM at 0 ° C to give the product in 91% yield.
  • a step of deprotection of the product in the presence of trifluoroacetic acid at room temperature provides the CDP-NH 2 , product 6, in a quantitative manner, this product is engaged in the next step without any purification.
  • the second part is the activation of paclitaxel with paranitrophenyl chloformate to give PTX-PNPh.
  • This reaction is carried out by successive addition of the formate for 4 h on a solution of paclitaxel in anhydrous dichloromethane at -50 ° C and in the presence of pyridine. After purification, PTX-PNPh, product 7, is obtained with a yield of 45%.
  • the third part consists in coupling the activated paclitaxel PTX-PNPh with the CDP-NH 2 in the presence of triethylamine at -20 ° C. in anhydrous DMF, the PTX-carbamate-CDP, product 8, is obtained with 51% yield .
  • This synthesis is carried out from PTX-PNPh and the RAFT CDP agent terminated by an alcohol function, CDP-OH.
  • the mixture is stirred for 48 h at room temperature in dry DCM and in the presence of a base (DMAP), the PTX-carbonate-CDP, product 9 is obtained with 65% yield.
  • paclitaxel-polyacrylamide prodrugs with different chemical bonds
  • the polyacrylamide prodrugs of paclitaxel with different chemical bonds were synthesized following the procedure described in Example 2 and using the synthesized RAFT agents: PTX-digly-CDP, PTX-carbamate-CDP and PTX-carbonate-CDP.
  • the following polymers are obtained: PTX-digly-PAAm M n 27,300, M w / M n 1,10; PTX-carbamate-PAAm M n 27,100, M w / M n 1.17; PTX-carbonate-PAAm M n 27.800, M w / M n 1.09.
  • Paclitaxel release experiments were performed in PBS (Tween 80, 1%) and in mouse plasma.
  • PTX, PTX-ester-PAAm (Example 2), PTX-digly-PAAm and PTX-carbonate-PAAm were incubated in PBS and murine plasma at 37 ° C. at the same concentration (1 mg / mL eq PTX).
  • PTX, 4h, 6h, 24h and 48h 200 ⁇ l of sample is taken for quantification.
  • Sample Preparation 200 ⁇ L aliquots are mixed with 600 ⁇ L of acetonitrile and 20 ⁇ L of deuterated Paclitaxel (PTX-d5) at 1 ⁇ g / mL (internal standard). Samples are shaken for 15 minutes and centrifuged for 10 minutes prior to analysis.
  • PTX-d5 deuterated Paclitaxel
  • ESI-MS / MS conditions The analyzes are performed on a triple quadrupole mass spectrometer detector (TQD) with electrospray ionization interface (ESI) (Quattro Ultima, Waters, Guyancourt, France). Electro spray and mass parameters were optimized by direct infusion of pure analytes into the system.
  • ESI parameters 3.5kV capillary voltage, 35V conical voltage, source temperature 120 0 C desolvation temperature 350 0 C, with a nitrogen flow rate of 506 L / h.
  • Mass parameters The transitions are monitored as follows PTX 854/286; PTX-d5 859/291.
  • the release profiles are illustrated in Figures 6 and 7.
  • Polymeric prodrugs with carbonate or ester bonds have better stability in PBS than with a diglycolate bond. The same trend is observed in murine plasma but the ester and carbonate bonds release quantitatively more active molecule (here PTX).
  • the synthesis is divided into different steps: in a first step, the peptide is coupled to a linker having a maleimide function, the trithiocarbonate function at the end of the polymer chain is then modified to give a thiol and the two elements are coupled by a thioether linkage .
  • the first step consists in coupling the targeting ligand to the linker by peptide coupling.
  • Cyclic RGD of cyclo (-Arg-Gly-Asp-D-Phe-Lys) sequence has a free amine from the residue of lysine which can be used for peptide coupling with the linker.
  • Cyclo-RGD trifluoroacetic salt (50.00 mg, 0.083 mmol) was dissolved in dry DCM (3 mL). DMF was added dropwise until solubilization of the solid. DIPEA (0.023 mL, 0.12 mmol) was added and the solution was stirred. In a separate flask, SMCC (27.70 mg, 0.083 mmol) was dissolved in dry DCM (2 mL) and the solution was added dropwise to the RGD solution. The resulting solution was stirred at room temperature for 24 h. The crude mixture was purified by preparative HPLC (50% H 2 0 / ACN in 15 minutes) and lyophilized to give a white powder (69.2 mg) 51% yield.
  • the second step consists of modifying the RAFT agent at the end of the polymer chain to give a thiol. This reaction is carried out by cleavage of the trithiocarbonate function of the CDP with an amine by aminolysis.
  • PTX-PAAm-CDP was dissolved in DMSO and the solution was degassed with argon for 10 min. Propylamine and n-tributylphosphine were added to the solution and stirred at room temperature for 48 h under an argon atmosphere. PAAm-SH was obtained as a powder after precipitation in cold diethyl ether. The powder was resuspended in DMSO and dialysed in Milli-Q water for 3 days. The solution was lyophilized to give a white solid, PTX-PAAm-SH.
  • the next step of synthesizing the thioether-bonded bioconjugate comprises coupling between the thiol of the maleimide linker with the free thiol of the previously obtained polymer.
  • the second synthetic route for the PTX-PAAm-RGD bioconjugate represents an alternative strategy for coupling the peptide to the polymer via a more stable peptide bond than the thioether linkage.
  • This synthesis consists first of all in modifying the trithiocarbonate group by a radical route to give a chain terminated by a carboxylic acid, and then coupling the targeting ligand by peptide coupling, according to for example the synthesis:
  • PTX-PAAm-CDP 300 mg, 0.015 mmol
  • ACPA 84 mg, 0.30 mmol
  • DMSO DMSO
  • PTX-PAAm-COOH (80.00 mg, 0.004 mmol) is then dissolved in water (8 mL). Then, NHS (0.5000 mg, 0.0048 mmol) and EDC.HCl (1500 mg, 0.008 mmol) are added. The mixture is stirred at t.a. for 24h. Then, cyclo-RGD trifluoroacetate (2.4 mg, 0.004 mmol) and DIPEA (1.4 ⁇ L, 0.008 mmol) are added and the mixture is stirred again for 24 hours. PTX-PAAm-RGD is obtained as a white powder after dialysis in Milli-Q water for 4 days and lyophilization (82% yield).
  • this polymer was carried out by the thiol-maleimide coupling method previously described. In a first step, rhodamine and cyanine were coupled to the maleimide linker, and then the coupling was carried out between the maleimide function and the thiol of the PTX-PAAm polymer obtained after aminolysis.
  • the inventors are, to their knowledge, the first to show that the prodrug polymer approach makes it possible to avoid the irritant / necrotizing effects of AP after SC injection.
  • Toxicity has been studied in mice. Increasing amounts of PTX (Taxol commercial formulation), PAAm (without PA), PTX-ester-PAAm (Example 2) and PTX-digly-PAAm (Example 8) were injected on D0. The weight of the mouse is followed, a weight loss of -10% is a sign of serious toxicity. No toxicity was observed with the various formulations tested. The results are shown in Figure 9.
  • Example 12 Study of Pharmacokinetics and Biodistribution (Radiolabelled PTX) Female BALB / c OlaHsd mice aged 7 weeks ( ⁇ 22 g, Envigo, France) were used. Taxol® radiolabeled and PTX * -PAAm radiolabeled (synthesized as in Example 1 with paclitaxel radiolabelled [3 H] -PTX) were injected at a dose of 1 mg.kg 7 (1 pCi per mouse) to perform pharmacokinetic and biodistribution studies.
  • mice were divided into four groups: (i) Taxol® injected intravenously; (ii) Taxol® injected subcutaneously; (iii) PTX-PAAm injected intravenously and (iv) PTX-PAAm injected subcutaneously.
  • Each group consisted of 40 mice divided into 10 different times (0.25 h, 0.5 h, 1 h, 2 h, 4 h, 4 h, 7 h, 24 h, 48 h, 96 h and 144 h) leading to 4 mice per group.
  • Radiolabelled PTX was added to the Taxol® formulation and radiolabeled PTX-PAAm was added to PTX-PAAm (approximately 1 pCi injected per mouse) to effect pharmacokinetics and biodistribution.
  • the mice were euthanized with pentobarbital and the blood was collected by cardiac puncture before the plasma was recovered by centrifugation in tubes containing EDTA (VACUETTE® K3 EDTA tube, 5 min centrifugation, 3000 ml). g). Livers, kidneys, rats, lungs and some SC tissues at the injection site were also collected. All the samples were stored in a freezer (-20 ° C.) for a maximum of one week before analysis.
  • Paclitaxel was radiolabelled and coupled to the polymer by an ester linkage (stable binding) so that it could be followed in the blood and in various organs (liver, lungs, kidneys, spleen, SC tissue).
  • ester linkage stable binding
  • Taxol is used as a control. At a dose of 7 mg / kg (paclitaxel equivalent), Taxol IV injected has a short half-life (a few tens of minutes).
  • Coupling to the polymer therefore increases the circulation time of paclitaxel by preventing its metabolism.
  • Biodistribution studies show predominantly hepatic elimination of paclitaxel. This is delayed for the paclitaxel polymer in accordance with pharmacokinetics. In the other organs, the quantities are small without disturbing accumulation.
  • the amounts of paclitaxel-polymer at the injection site decrease rapidly, which confirms the rapid passage of the prodrug into the circulation and the good bioavailability of the polymer-Paclitaxel.
  • the present invention therefore makes it possible to increase the solubility, the high concentration stability and bioavailability of the active ingredient.
  • the polyacrylamide was labeled with a fluorescent probe (Rhodamine, Example 6) and the fluorescence followed in vivo in the mouse by means of a Lumina imaging system (PerkinElmer) with excitation filters between 500 and 535 nm, and the filters emission between 575 and 650 nm.
  • a fluorescent probe Rhodamine, Example 6
  • Lumina imaging system PerkinElmer
  • Free rhodamine injected SC has good bioavailability (fluorescence decreases in 24 hours at the injection site) and is rapidly eliminated.
  • Rhodamine-PAAm is injected IV, rhodamine is still detectable 4 days after injection. Polymer grafting protects the probe against metabolism and excretion, the half-life is greatly increased.
  • the rhodamine-SC polymer is bioavailable and has an increased circulation time (still present at 4 days). These results show that coupling to the polymer increases the circulation time of the active molecule.
  • the present invention is therefore advantageous for slowing down the elimination of the active ingredients once coupled.
  • the diglycolate bond which releases the PTX more rapidly makes it possible to have higher concentrations in the course of time.
  • the ester bond which liberates more slowly allows for a prolonged release.
  • the carbonate bond weakly releases.
  • the following example shows the synthesis of a polymer having, as grafted active molecule, a polar molecule. Protection of the hydroxyl functions of Gemcitabine
  • the transmittance results as a function of temperature are summarized in Table 2.
  • the UCST is determined as the temperature where the transmittance reaches about 100%, ie when the polymer is fully solubilized.
  • the polymer is dialyzed against 1 L of water for 24 hours. The water is changed in the day every 4 hours. At the end of the dialysis, the polymer is lyophilized. White pasty flakes are obtained. The 1 H-NMR spectrum of the product obtained confirms the presence of the polymerization of
  • a PEGMA chain with a length of 1400 to 2000 g / mol was added.
  • CP5-PEGMA and CP7-PEGMA were analyzed by UV-visible spectrometer to determine their UCST as 411 ° C and 52 ° C respectively with a low hysteresis (of the order of 1 to 2 ° C).
  • Viscosity protocol (depending on the shear rate):
  • mice with PTX-21% -5 A local toxicity test is performed in mice with PTX-21% -5. On a group of 3 "nude" mice, 0.6 mg of PTX in 200 ⁇ l of PBS solution are administered subcutaneously. This injection is repeated 4 times over 4 consecutive days. At the end of the third day, local toxicity (acute irritation, mild necrosis) occurred near the injection site for all mice, as shown in Figure 3.

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