WO2025031991A1 - Lipo-polyaminoacides contenant du soufre stabilisés et furitfs - Google Patents

Lipo-polyaminoacides contenant du soufre stabilisés et furitfs Download PDF

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WO2025031991A1
WO2025031991A1 PCT/EP2024/072025 EP2024072025W WO2025031991A1 WO 2025031991 A1 WO2025031991 A1 WO 2025031991A1 EP 2024072025 W EP2024072025 W EP 2024072025W WO 2025031991 A1 WO2025031991 A1 WO 2025031991A1
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formula
cio
lipo
moiety
alkyl
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Inventor
Timothy Jon DEMING
Carles FELIP LEON
Josep GARCIA GARCIA
Vicent Josep Nebot Carda
Lidia HERRERA MUÑOZ
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Polypeptide Therapeutic Solutions SL
University of California Berkeley
University of California San Diego UCSD
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Polypeptide Therapeutic Solutions SL
University of California Berkeley
University of California San Diego UCSD
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    • A—HUMAN NECESSITIES
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    • 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
    • A61K47/69—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
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    • A61K47/6935—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
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    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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    • 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
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    • 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
    • A61K47/54—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 compound
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • A—HUMAN NECESSITIES
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    • 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
    • A61K47/54—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 compound
    • A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
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    • 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
    • A61K47/54—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 compound
    • A61K47/55—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug
    • A61K47/551—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
    • 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
    • A61K47/62—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 a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • 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
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • 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
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/5123—Organic compounds, e.g. fats, sugars
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G69/00—Macromolecular compounds obtained by reactions forming a carboxylic amide link in the main chain of the macromolecule
    • C08G69/02—Polyamides derived from amino-carboxylic acids or from polyamines and polycarboxylic acids
    • C08G69/08—Polyamides derived from amino-carboxylic acids or from polyamines and polycarboxylic acids derived from amino-carboxylic acids
    • C08G69/10—Alpha-amino-carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle

Definitions

  • the present disclosure relates to lipo-polyamino acid conjugates which comprise a sulfur-containing polyamino acid (PAA) moiety conjugated to a lipid-like moiety.
  • PAA sulfur-containing polyamino acid
  • These lipo-polyamino acid conjugates may form self-assembled nanoparticles, such as for example lipid nanoparticles (LNPs), and can be used as non- viral vectors for delivery of active ingredients, including nucleic acids, to cells.
  • LNPs lipid nanoparticles
  • Nucleic acids have emerged in recent years to yield promising drug candidates for a wide range of diseases, including, among others, cancer, infectious diseases, and cardiovascular, inflammatory, and neurodegenerative diseases. Nucleic acids exert their action via gene inhibition, addition, replacement or editing. However, their clinical efficacy is very much limited because of their inability to cross physiological barriers. The negative charge and hydrophilicity of nucleic acids hinder their passive diffusion across plasma membranes. Moreover, the association of nucleic acids with serum proteins as well as their susceptibility to enzymatic degradation by endogenous nucleases also interferes with their efficient translation to functional response.
  • Non-viral vectors such as LNPs are known to permeate plasma membranes and thus to be able to deliver nucleic acids to cells.
  • LNPs are vesicles composed of mixtures of lipids that are physically associated with each other by intermolecular forces forming a micellar structure. LNPs typically have a small diameter of less than 200 nm. LNPs are used as drug delivery system thanks to their ability to encapsulate biologically active agents and deliver them to specific locations within the body at a desired time.
  • the lipid components of LNPs may include ionizable or cationic lipids, helper lipids such as phospholipids and structural lipids such as sterols or cholesterol. lonizable/cationic lipids, which have either a permanent or ionizable positively charged head group followed by a hydrophobic tail, can form complexes with negatively charged nucleic acids by means of electrostatic bonds.
  • LNPs One of the potential problems of LNPs is that once they are injected, they may be recognised by the immune system as foreign particles and subsequently cleared from the bloodstream by phagocytes.
  • different strategies may be used including decreasing the particle size of the vesicles or modifying their surface.
  • the LNP surface may be modified by conjugating the hydrophilic polymer polyethylene glycol (PEG) to lipids such as phospholipids or long-chain fatty acids to form PEG-lipids.
  • PEG is a hydrophilic polymer widely used in drug delivery and nanotechnology because of its stealth properties and biocompatibility. Thanks to these properties, particulate delivery systems containing PEG are able to evade the immune system, and, as a result, have a prolonged circulation time within the body.
  • hydrophilic polymers such as poly (vinylpyrrolidones), poly (acryloyl-morpholines) or polyoxazolines have been reported as alternatives to PEG-lipids, no LNP containing such polymers have been marketed yet.
  • Polysarcosine is a polypeptoid based on the endogenous amino acid sarcosine (N-methylated glycine), which has previously shown potent stealth properties.
  • Lipid nanoparticles (LNPs) for therapeutic application of messenger RNA assembled using pSarcosinylated lipids have been reported as a promising tool for particle engineering (ACS Appl. Nano Mater., 2020, 3, 10634—10645) but this is still under development and no clinical data including this polymer into LNPs is presented yet.
  • the inventors have developed new lipo-polyamino acid conjugates which comprise a sulfur-containing polyamino acid (PAA) moiety conjugated to at least one lipid-like moiety. These conjugates may form selfassembled nanoparticles, such as for example lipid nanoparticles (LNPs), and can be effectively used as non- viral vectors for delivery of active ingredients, including nucleic acids, to cells, thanks to their stealth properties and consequent improved plasma half-life time.
  • PAA sulfur-containing polyamino acid
  • LNPs lipid nanoparticles
  • the sulfur-containing lipo-polyamino acid conjugates show low cytotoxicity, high biodegradability and may confer to the final nanosystem enhanced efficiency, high stability due to limited or even completely suppressed aggregation issues in the bloodstream and potential different cell and tissue tropism.
  • a first aspect of the invention relates to a lipo-polyamino acid conjugate of formula (I), a salt thereof, or any stereoisomer or mixtures of stereoisomers, either of the compound of formula (I), or of any of its salts, wherein:
  • PAAi is a repetitive unit of formula (II)
  • PAA2 is a repetitive unit of formula (III) wherein though the repeating units PAA1 and PAA2 are shown in a particular order for convenience of description, the repeating units may be present in any order and may be block or randomly present; and wherein each of the repeating units PAA1 and PAA2 may comprise blocks of monomer units which may be the same or different between each other; n is an integer from 5 to 250; m is an integer from 0 to 250; p is 0 or 1; s Is O or 1 ;
  • X is selected from the group consisting of CH, N, S, and 0;
  • X' is selected from the group consisting of N and 0;
  • R1 and R1' are independently selected from the group consisting of H, -(Ci-Ci8)alkyl, and -(C2-Ci8)alkenyl, with the condition that R1 is absent when X is 0 or S, and R is absent when X' is 0;
  • R2 is a radical selected from the group consisting of (IV), (V) and (VI)
  • R3 and R5 are independently selected from the group consisting of H and -(Ci-Cejalky I;
  • R4 is any amino acid side chain optionally functionalized with an active moiety
  • A is selected from the group consisting of
  • A1 is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, -(C1-C10) alkoxy, -(C6-Cio)aryloxy, -(C6-Cio)aralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alkyl-0-(C6-Cio)aryloxy, an amino protective group, a lipid- like moiety R , an amino acid-like moiety Rs, and an active moiety; with the condition that:
  • A1 is selected from the group consisting of H, -(Ci-Cejalkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, an amino protective group, a lipid-like moiety R , an amino acid-like moiety Rs, and an active moiety; ii) when p is 1 and s is 0, A1 is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-
  • a and Ai are independently optionally substituted by one or more groups selected from the group consisting of -OH, halogen, -CF3, -NH2, -NH-(Ci-C4)alkyl, -NH-CO-(Ci-C6)alkyl, -(Ci-Cejalkyl, -NO2, -N3, -CO-(Ci-C 6 )alkyl, -CO-O-(Ci-C 6 )alkyl, -SO 3 H, -SO2NH2, -SO 2 -N((Ci-C 6 )alkyl) 2 , -COOH, CONH 2 , and -CON((Ci-C 6 )alkyl) 2 ; each lipid-like moiety Rz is independently selected from the group consisting of -(Ci-C2o)alkyl, -(C2-C2o)alkenyl, each Rz' is independently selected from the group consisting of -
  • Y and Y' are independently selected from -OH, -OCORx and -COORx;
  • Q and Q' are independently selected from -OCORy and -COORy;
  • Z and Z' are independently selected from -O-, -OCO-, -COO-, -NRz'CO-, and -CONRz'-; each Rz' is H or Rz; each Rx, Ry, and Rz is independently -(Ci-Ci8)alkyl or -(C2-Ci8)alkenyl; each g is independently an integer from 0 to 18; each h is independently an integer from 1 to 18;
  • R22, R33 and R44 are independently selected from the group consisting of hydrogen, fluorine, methyl, -CH2F, -CHF2, and -CF3; each of the dashed bonds — is independently a single bond or, alternatively, a double bond;
  • a second aspect of the invention relates to a lipo-polyamino acid conjugate of formula (I), a salt thereof, or any stereoisomer or mixtures of stereoisomers, either of the compound of formula (I), or of any of its salts, wherein the lipo-polyamino acid conjugate of formula (I) is as defined in the first aspect wherein:
  • a and Ai may also be independently optionally substituted by one or more groups selected from the group consisting of -(CHg -SO-CHg, and NH(Ci-Ce)alkyl; and
  • Ai is other than a lipid-like moiety Rz of formula (XVII) wherein each j is16 and i is 2.
  • the lipo-polyamino acid conjugates of the present disclosure may be decorated with cell-targeting groups in any of A, Ai, A2 or A3 that can actively target cells and aid in cellular entry, resulting in a conjugate which has improved cell-specific delivery.
  • the lipo-polyamino acid conjugates may also be decorated with penetration enhancing agents when an increased cell or skin penetration is desired, Further, the lipo-polyamino acid conjugates can also be used for delivering pharmaceutically, cosmetically or diagnostically active agents attached to the backbone of the conjugate in any of A, Ai, A2 or A3.
  • the lipo-polyamino acid conjugates of the invention may form self-assembled particles in solution such as for example liposomes or lipid nanoparticles which may be used to encapsulate and deliver molecules of a different nature.
  • the lipo-polyamino acid conjugates of the invention allow carrying and/or delivering different active agents at the same time, not only bonded to the structure of the conjugate but also contained in self-assembled particles formed from the conjugated.
  • another aspect of the invention relates to a self-assembled particle comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, and optionally one or more active agents selected from the group consisting of pharmaceutically active agents, cell-targeting agents, penetration enhancing agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • the lipo-polyamino acid conjugates of the invention may be formulated in a variety of compositions, including pharmaceutical, cosmetic and diagnostic compositions, with excipients and carriers.
  • another aspect of the invention relates to a composition comprising the lipo-polyamino acid conjugate as defined herein; or alternatively, the self-assembled particle containing them, together with one or more appropriate excipients or carriers.
  • the lipo-polyamino acid conjugates of the invention may be used in medicinal, cosmetic and diagnostic applications.
  • a therapeutic product which is or comprises: a) a lipo- polyamino acid conjugate of formula (I) as defined herein, wherein at least one of A, A1, A2 or A3 is a pharmaceutically active agent; or alternatively, b) a self-assembled particle containing the lipo-polyamino acid conjugate a); or alternatively, c) a composition containing the lipo-polyamino acid conjugate a) or the selfassembled particle b); or alternatively, d) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more active agents selected from the group consisting of pharmaceutically active agents, nucleic acids, peptides, proteins, and mixtures thereof; or alternatively, e) a composition containing the self-assembled particle d), for use in medicine.
  • the invention relates to a diagnostic product which is or comprises: a') a lipo- polyamino acid conjugate as defined herein, wherein at least one of A, A1, A2 or A3 is a diagnostically active agent; or alternatively, b') a self-assembled particle containing the lipo-polyamino acid conjugate a'); or alternatively, c') a composition containing the lipo-polyamino acid conjugate a') or the self-assembled particle b'); or alternatively, d) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more diagnostically active agents; or alternatively, e') a composition containing the self- assembled particle d'), for use in diagnostics.
  • the invention relates to the use in cosmetics of a cosmetic product which is or comprises: a”) a lipo-polyamino acid conjugate as defined herein, wherein at least one of A, Ai, A2 or A3 is a cosmetically active agent; or alternatively, b”) a self-assembled particle containing the lipo-polyamino acid conjugate a'); or alternatively, c”) a composition containing a”) or the self-assembled particle b”); or alternatively, d”) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more cosmetically active agents; or alternatively, e”) a composition containing the selfassembled particle d”).
  • Another aspect of the invention relates to the use of the lipo-polyamino acid conjugate of formula (I) as defined herein, wherein A, A1, A2 and A3 are other than an active moiety, as a carrier.
  • FIG. 1 shows an agarose gel (2%) for the detection of free mRNA from different LNPs formulations: M) free mRNA; 1) benchmark LNPs formulation; 2) benchmark LNPs formulation + 1% triton X-100; 3) PMetO5 LNPs formulation; 4) PMetO5 LNPs formulation + 1% triton X-100.
  • FIG 2 shows an agarose gel (2%) for the detection of free mRNA from different LNPs formulations: M) free mRNA; 1) benchmark LNPs formulation; 2) benchmark LNPs formulation + 1% triton X-100; 3) PMetOI LNPs formulation; 4) PMetOI LNPs formulation + 1% triton X-100.
  • moiety refers to a specific segment or functional group of a molecule or compound.
  • the term "subject” refers to any mammal, including both human and other mammals.
  • the term -(Ci-C x )alkyl refers to a saturated linear or branched hydrocarbon chain which contains from 1 to x carbon atoms and only single bonds. Examples of alkyl groups may include without limitation methyl, ethyl, n- propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, neopentyl, n-hexyl, decyl, undecyl, dodecyl, tetradecyl and hexadecyl.
  • alkenyl refers to an unsaturated branched or linear hydrocarbon chain which comprises from 2 to x carbon atoms and at least one or more double bonds.
  • alkenyl groups may include without limitation ethenyl (i.e. vinyl), allyl, propenyl, butenyl, pentenyl and hexenyl, dodecenyl, tetradecenyl and hexadecenyl.
  • alkoxy refers to an alkyl group as defined herein comprising 1 to 10 carbon atoms which is connected to the rest of the molecule via an oxygen atom.
  • alkoxy groups may include without limitation methoxy, ethoxy, 1 -propoxy, and 2-propoxy.
  • -(C5-Cio)aryl refers to an aromatic carbocyclic mono- or bicyclic ring system comprising 5 to 10 carbon ring atoms.
  • aryl groups may include without limitation phenyl, biphenyl, and naphthyl.
  • -(C6-Cio)aryloxy refers to an aryl group as defined herein comprising 6 to 10 carbon ring atoms, which is connected to the rest of the molecule via an oxygen atom.
  • aryloxy groups may include without limitation phenoxy and biphenyloxy.
  • -(C5-Cio)heteroaryl refers to an aromatic carbocyclic mono- or bicyclic ring system comprising 5 to 10 ring atoms, wherin at least one aromatic ring containing one, two, or three ring heteroatoms selected from N, O, and S, the remaining ring atoms being carbon atoms.
  • heteroaryl groups may include without limitation pyridyl, azetidinyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl, isoquinolinyl, benzthiazolyl, benzimidazolyl, benzthiazolyl, quinolyl, and quinazolyl.
  • -(C6-Cio)aralkyl refers to a lower alkyl group as defined herein (e.g. -(Ci-Ce)alkyl) which is substituted by an aryl group (e.g. “-(C5-Cio)aryl) as defined herein.
  • aralkyl groups may include without limitation benzyl, phenethyl, and methylbenzyl.
  • -(C6-Cio)aralkoxy refers to a lower alkoxy group as defined herein (e.g. -(Ci-Ce)alkoxy) which is substituted by an aryl group (e.g. “-(C5-Cio)aryl) as defined herein.
  • aralkoxy groups may include without limitation benzyloxy, phenethoxy, and methylbenzyloxy.
  • heteroaralkoxy refers to a lower alkoxy group as defined herein (e.g. -(Ci-Ce) alkoxy) which is substituted by an heteroaryl group (e.g. "-(C5-Cio)heteroaryl) as defined herein.
  • heteroaralkoxy groups may include without limitation furyloxy, pyridyloxy, and azetidinyloxy.
  • heterocycloalkyl refers to a 5-10 membered mono- or bicylic (fused or bridged) saturated o unsaturated ring structure, in which one or more of the ring atoms is a heteroatom selected from N, O, and S.
  • heterocycloalkyl groups may include without limitation include piperidyl, piperazyl, tetrahydropyranyl, tetrahydrofuryl, 1,4-dioxolanyl, oxazolidyl, isoxazolidyl, morpholinyl, thiomorpholyl.
  • protecting group refers to a grouping of atoms that when attached to a reactive group in a molecule masks, reduces or prevents that reactivity.
  • substituted means that one or more hydrogen atoms on the designated atom or group are replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded. Combinations of substituents and/or variables are permissible.
  • optionally substituted means that the number of substituents can be equal to or different from zero. Unless otherwise indicated, it is possible that optionally substituted groups are substituted with as many optional substituents as can be accommodated by replacing a hydrogen atom with a non-hydrogen substituent on any available carbon or nitrogen atom. It is possible that groups in the conjugates according to the invention are substituted with one, two, three, four or five identical or different substituents, particularly with one, two or three substituents.
  • a first aspect of the invention refers to a lipo-polyamino acid conjugate of formula (I), a salt thereof, or any stereoisomer or mixtures of stereoisomers, either of the compound of formula (I), or of any of its salts.
  • A, X, X', Ri, R/, n, m, p, s and Ai are as defined herein, and PAAi and PAA2 are repetitive units of formulas (II) and (III), respectively
  • repeating units PAAi and PAA2 are shown in a particular order for convenience of description, the repeating units may be present in any order and may be block or randomly present; and wherein each of the repeating units PAA1 and PAA2 may comprise blocks of monomer units which may be the same or different between each other.
  • I ipo-poly amino acid conjugate refers to a molecule which comprises a) at least one polyamino acid-based moiety and b) at least one lipid-like moiety.
  • lipid- like moiety refers to a moiety which, while it cannot be considered to be a lipid as such, structurally and/or functionally resembles a lipid.
  • lipid-like moiety is intended to include moieties that are able to form amphiphilic layers.
  • lipid refers to a group of organic compounds such as fatty acids, glycerolipids, glycerophospholipids, sphingolipids, saccharolipids, sterols, and the like, which are insoluble in water.
  • the lipid-like moiety can be placed on any of the terminal ends of the (PAAi) n -(PAA2) m -based moiety. It is also possible that the lipo-polyamino acid conjugates of the invention comprise a lipid-like moiety at both ends of the molecule.
  • the lipid like moiety can be placed at the left side of the drawn molecule of formula (I) when A is R7, or at the right side when A1 is R , or at both ends when A is R7, and A is R .
  • at least one of A and A1 must be a lipid-like moiety (R7 in the case of A or R in the case of A1) .
  • such salts can be prepared by reacting the free acid or base form of a lipo-polyamino acid conjugate of the invention with a stoichiometric amount of an appropriate base or acid, respectively, in a suitable solvent such as water, an organic solvent or a mixture of them.
  • suitable solvent such as water, an organic solvent or a mixture of them.
  • pharmaceutically, cosmetically or diagnostically acceptable salts include acid addition salts formed with inorganic acids e.g. hydrochloric, hydrobromic, sulfuric, nitric, hydroiodic, metaphosphoric, or phosphoric acid; and organic acids e.g.
  • succinic maleic, acetic, fumaric, citric, tartaric, benzoic, trifluoroacetic, malic, lactic, formic, propionic, glycolic, gluconic, camphorsulfuric, isothionic, mucic, gentisic, isonicotinic, saccharic, glucuronic, furoic, glutamic, ascorbic, anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic), ethanesulfonic, pantothenic, stearic, sulfinilic, alginic and galacturonic acid; and arylsulfonic, for example benzenesulfonic, p-toluenesulfonic, oxalic, methanesulfonic or naphthalenesulfonic acid; and base addition salts formed with alkali metals and alkaline earth metals and organic bases such as N,N- dibenz
  • stereoisomer refers to all isomers of individual lipo- polyamino acid conjugates that differ only in the orientation of their atoms in space.
  • stereoisomer includes mirror image isomers (enantiomers), mixtures of mirror image isomers (racemates, racemic mixtures), geometric (cis/trans or syn/anti or E/Z) isomers, and isomers of lipo-polyamino acid conjugates with more than one chiral center that are not mirror images of one another (diastereoisomers).
  • the present invention relates to each of these stereoisomers and also mixtures thereof.
  • the definition of the lipo- polyamino acid conjugates of formula (I) is also intended to encompass all R- and S-isomers of a chemical structure in any ratio, e.g. with enrichment (i.e. enantiomeric excess or diastereomeric excess) of one of the possible isomers and corresponding smaller ratios of other isomers.
  • enrichment i.e. enantiomeric excess or diastereomeric excess
  • they may acquire L-configuration or D-configuration.
  • Diastereoisomers and enantiomers can be separated by conventional techniques such as chromatography or fractional crystallization. Optical isomers may be obtained using enantiospecific synthesis. Alternatively, optical isomers can be resolved by conventional techniques to give optically pure isomers. The resolution can be carried out on any chiral synthetic intermediates or on the polymers of the invention.
  • the pharmaceutically, cosmetically or diagnostically acceptable salts thereof and the stereoisomers or mixtures of stereoisomers either of any of the lipo-polyamino acid conjugates or of any of their pharmaceutically, cosmetically or diagnostically acceptable salts are always contemplated even if they are not specifically mentioned.
  • the lipo-polyamino acid conjugates of the invention may be in crystalline form either as free solvation lipo- polyamino acid conjugates or as solvates (e.g. hydrates). It is intended that all these forms are within the scope of the present invention. Methods of solvation are generally known within the art. For the purposes of the invention, the solvated forms with pharmaceutically, cosmetically or diagnostically acceptable solvents such as water, ethanol and the like are equivalent to the unsolvated form.
  • the lipo-polyamino acid conjugates of the present disclosure also include lipo- polyamino acid conjugates that differ only in the presence of one or more isotope-enriched atoms.
  • isotope-enriched atoms without limitation, are deuterium, tritium, 13 C or 14 C, or a nitrogen atom enriched in 15 N or a fluorine atom enriched in 18 F.
  • the lipo-polyamino acid conjugate of formula (I) comprise “n” PAAi repetitive units of formula (II) and optionally “m” PAA2 repetitive units of formula (III).
  • the weight ratio of the PAA1 monomer to the PAA2 monomer in the conjugate is from 100/0 to 50/50, more particularly from 100/0 to 75/25.
  • the term “homopolymer” refers to a polymer derived from a single monomer.
  • copolymer refers to a polymer derived from more than one monomer.
  • the copolymer may be a random or a block copolymer.
  • random copolymer refers to a copolymer in which the monomer units are located randomly in the polymer molecule.
  • block copolymer refers to a copolymer that comprises at least two different monomer units that upon polymerization form at least two chemically distinct regions, segments or blocks that are chemically distinguishable from one another.
  • block copolymer includes linear block copolymers, multiblock copolymers and star shaped block copolymers.
  • the lipo-polyamino acid conjugates of the invention are characterized by a molecular weight, which can be average molecular weight (MW) or number average molecular weight (Mn), a degree of polymerization, and a polydispersity index.
  • the molecular weight can be measured by methods well-known in the art such as size exclusion chromatography (SEC) (also referred to as gel permeation chromatography (GPC)) matrix assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS).
  • SEC size exclusion chromatography
  • GPC gel permeation chromatography
  • MALDI-TOF MS matrix assisted laser desorption ionization-time-of-flight mass spectrometry
  • the term "repeating unit” refers to the monomeric amino acid-based unit and is defined by square brackets ("[ ]”) depicted around it.
  • the number on the lower right of the brackets (represented by letters “n” or “m”) refers to the degree of polymerization (DP) for each monomeric unit as a statistical number.
  • the DP is calculated by dividing the corrected molecular weight (subtracting the initiator MW) of the polymer determined by SEC-MALS technique by the molecular weight of the monomer unit.
  • the DP value is reported as the central value of a gaussian distribution which comprises polymers of variable DP (depending on intrinsic polydispersity).
  • the DP value is subject to a reasonable uncertainty, due to the ring-opening polymerization mechanism, which, in the context of the present invention, may be considered within the range ⁇ 20%, preferably ⁇ 15%, more preferably ⁇ 10%, even more preferably ⁇ 5%, being particularly preferred ⁇ 2%.
  • polydispersity index (PD I ) is used as a measure of broadness of molecular weight distribution. The larger the PDI, the broader the molecular weight. PDI of a polymer is calculated as the ratio of weight average (MW) by number average (Mn) molecular weight.
  • the lipo-polyamino acid conjugate of formula (I) is a copolymer comprising “n” PAAi repetitive units of formula (II) and “m” PAA2 repetitive units of formula (III), more particularly wherein the weight ratio of the PAA1 monomer to the PAA2 monomer in the conjugate is from 95/5 to 50/50, for example about 10/5, about 20/5, about 30/5, about 40/5, about 50/5, about 60/5, about 70/5 or about 80/5, particularly measured 1 H-NMR spectroscopy. More particularly, the copolymer is a random or a block copolymer.
  • the lipo-polyamino acid conjugate has the formula (IB): wherein A, A1, X, X', R1, R/, R2, R3, R4, R5, n, m, p and s are as defined herein.
  • the lipo-polyamino acid conjugate of formula (I) is a homopolymer comprising “n” PAA1 repetitive units of formula (II), i.e., wherein PAA2 repetitive units of formula (III) are absent.
  • the lipo-polyamino acid conjugate according has the formula (IA) wherein A, A1, X, X', R1, R/, R2, R3, n, and p and s are as defined herein.
  • the lipo-polyamino acid conjugate of formula (I) has the formula (IA'): wherein A, Ai, X, X', Ri, R/, R2, R3, n, and p and s are as defined herein.
  • n and m are independently an integer from 5 to 250, particularly measured by 1 H-NMR spectroscopy.
  • n is an integer from 5 to 150, more particularly from 5 to 100, even more particularly from 5 to 50, and even more particularly from 5 to 30 or from 5 to 25. In another embodiment, optionally in combination with any of the embodiments provided above or below, n is a value from 5 to 200, from 10 to 200, from 10 to 150, or from 10 to 100.
  • m is an integer from 5 to 150, more particularly from 5 to 100, even more particularly from 5 to 50, and even more particularly from 5 to 30 or from 5 to 25.
  • n is a value from 5 to 200, from 5 to 80, from 10 to 80, from 5 to 70, from 10 to 70, from 5 to 60 or from 10 to 60.
  • n and m are independently about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 10, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 210, about 220, about 230, about 240, or about 250, particularly measured by 1 H-NMR spectroscopy.
  • X is selected from CH, N, S, and O. In one embodiment, optionally in combination with any of the embodiments provided above or below, X is N and R1 is H. In another embodiment, optionally in combination with any of the embodiments provided above or below, X is N and R1 is -(Ci-Cisjalkyl, more particularly -(Ci-Ci2)alkyl, even more particularly -(Ci-Cejalkyl.
  • X is N and R1 is -(C2-Ci8)alkenyl, more particularly -(C2-Ci2)alkenyl, even more particularly -(C2-C6)alkenyl.
  • X is 0 and Ri is absent.
  • X is S and Ri is absent.
  • X is CH and Ri is H.
  • p and s are 0 or 1 depending on the meaning of Ai.
  • p is 0 and s is 0, and Ai is selected from the group consisting of H, -(Ci-Cejalkyl, -(Cs-C jaryl, -(Cs-Ciojheteroaryl, -(Ce-Ciojaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(Cs-Ciojheterocycloalkyl, an amino protective group, a lipid-like moiety R , an amino acid-like moiety Rs, and an active moiety, wherein and Ai is as defined herein.
  • p is 1 and s is 0, and Ai is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(Cs-C jaryl, -(Cs-Ciojheteroaryl, -(Ce-Ciojaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(Cs-Ciojheterocycloalkyl, -(Ci-C jalkoxy, -(Ce-Cwjaryloxy, -(C6-Cio)aralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alkyl-0-(C6-Cio)aryloxy, a lipid-like moiety R , and an active moiety, wherein and Ai is as defined herein.
  • p is 1 and s is 1, and Ai is selected from the group consisting of H, -(Ci-Cejalkyl, -(Cs-C jaryl, -(Cs-Ciojheteroaryl, -(Ce-Ciojaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(Cs-Ciojheterocycloalkyl, -(Ci-Cio)alkoxy, -(Ce-Cwjaryloxy, -(Ce-Cwjaralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alkyl-0-(C6-Cio)aryloxy, a lipid-like moiety R , and an active moiety, wherein and Ai is as defined herein.
  • X' is selected from N and 0.
  • s is 1, X' is N and Ri' is H.
  • s is 1, X' is N and R/ is -(Ci-Cisjalkyl, more particularly -(Ci-Cejalkyl even more particularly -(Ci-Cejalkyl.
  • X' is N and R/ is -(C2-Ci8)alkenyl, more particularly -(C2-Ci2)alkenyl, even more particularly -(C2-C6)alkenyl.
  • s is 1, X' is 0 and R/ is absent.
  • R2 is a radical selected from the group consisting of (IV) and (V), wherein a and b are independently an integer from 0 to 3; and R is -(Ci-C6)alkyl.
  • R2 is a radical of formula (IV) and a is 0. In another embodiment, optionally in combination with any of the embodiments provided above or below, in the lipo-polyamino acid conjugate of formula (I), R2 is a radical of formula (IV) and a is 1. In another embodiment, optionally in combination with any of the embodiments provided above or below, in the lipo-polyamino acid conjugate of formula (I), R2 is a radical of formula (IV) and a is 2. In another embodiment, optionally in combination with any of the embodiments provided above or below, in the lipo-polyamino acid conjugate of formula (I), R2 is a radical of formula (IV) and a is 3.
  • the radical of formula (IV) is selected from the formulas (iv1)-(iv24):
  • R2 is a radical of formula (V) and b is 0.
  • R2 is a radical of formula (V) and b is 1.
  • R2 is a radical of formula (V) and b is 2.
  • R2 is a radical of formula (V) and b is 3.
  • R2 is a radical of formula (VI) and c is 0.
  • R2 is a radical of formula (VI) and c is 1 .
  • R2 is a radical of formula (VI) and c is 2.
  • R2 is a radical of formula (VI) and c is 3.
  • radical of formula (VI) is selected from the formulas (vi1)-(vi72):
  • R3 is H or -(Ci-Cejalkyl. In one embodiment, optionally in combination with any of the embodiments provided above or below, in the lipo-polyamino acid conjugate of formula (I), R3 is H. According to another embodiment, optionally in combination with any of the embodiments provided above or below, in the lipo-polyamino acid conjugate of formula (I), R3 is -(Ci-Cejalkyl, more particularly R3 is -CH3.
  • R4 is any amino acid side chain optionally functionalized with an active moiety. According to another embodiment, optionally in combination with any of the embodiments provided above or below, R4 is any amino acid side chain.
  • amino acid refers to a molecule containing both an amino group and a carboxyl group attached through a carbon atom to which a side chain is attached.
  • Suitable amino acid side chains in R4 include, without limitation, the side chains of alpha amino acids, such as the L-isomers of alpha-amino acids of the 20 common naturally occurring alpha-amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine; natural betaamino acids (e.g., beta-alanine); and unnatural amino acids.
  • alpha amino acids such as the L-isomers of alpha-amino acids of the 20 common naturally occurring alpha-amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine
  • unnatural amino acid comprises D-isomers of the 20 common naturally occurring alpha-amino acids as defined above or other amino acids as defined in Table A.
  • unnatural amino acids include 2-Aminoadipic acid (Aad), 3- Aminoadipic acid (bAad), beta-Alanine, beta-Aminopropionic acid (bAla), 2-Aminobutyric acid (Abu), 4- Aminobutyric acid, piperidinic acid (4Abu), 6-Aminocaproic acid (Acp), 2-Aminoheptanoic acid (Ahe), 2- Aminoisobutyric acid (Aib), 3-Aminoisobutyric acid (bAib), 2-Aminopimelic acid (Apm), 2,4 Diaminobutyric acid (Dbu), Desmosine (Des), 2,2’-Diaminopimelic acid (Dpm), 2,3-Diaminopropionic acid (Dpr), N-Ethylglycine (EtGly), N-Ethylasparagine (EtAsn), Hydroxylysine (Hyl), all
  • Each one of the amino acids forming the peptide of the invention can have, independently from the others, L- or D-configuration.
  • R4 corresponds to the amino acid the side chains of phenylalanine, alanine, leucine, glycine, isoleucine, or valine, more particularly R4 corresponds to the amino acid the side chain of alanine.
  • R5 is H or -(Ci-Ce) alkyl. In one embodiment, optionally in combination with any of the embodiments provided above or below, R5 is H. According to another embodiment, optionally in combination with any of the embodiments provided above or below, R5 is -(Ci-C6)alkyl, more particularly R5 is -CH3.
  • A is selected from the group consisting of H, -(Ci-C6)alkyl, -CO(Ci-C6)alkyl, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(Ce-
  • A1 is selected from the group consisting of H, -(Ci-Cejalkyl, -(Cs-Ciojaryl, -(Cs-Ciojheteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, an amino protective group, a lipid-like moiety R , an amino acid-like moiety Rs, and an active moiety; ii) when p is 1 and s is 0, A1 is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(Cs-Ciojaryl, -(Cs-Ciojheteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C
  • one of A and A1 is a lipid-like moiety Rz or Rz' as defined herein, and the other one of A and A1 is other than a lipid-like moiety Rz or R , respectively.
  • A is a lipid-like moiety Rz
  • A1 is other than a lipid-like moiety R ; more particularly, A1 is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(Cs-Ciojaryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, -(C1-C10) alkoxy, -(C6-Cio)aryloxy, -(C6-Cio)aralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alky
  • A1 is a lipid-like moiety Rz', and A is other than a lipid-like moiety Rz; more particularly, A is selected from the group consisting of A is selected from the group consisting of H, -(Ci-Cejalkyl, -CO(Ci-C6)alkyl, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(Ce- Cio)aralkyl,
  • the lipid-like moiety Rz in A in the lipo-polyamino acid conjugate of formula (I) is selected from the group consisting of -(Ci-C2o)alkyl, -(C2-C2o)alkenyl, and a radical of formula (IX), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII).
  • the lipid- like moiety Rz is -(Ci-C2o)alkyl, more particularly -(Ce-Cisjalkyl, and even more particularly -(Ci2-Cis)alkyl.
  • lipid-like moiety Rz has the formula (XXIII):
  • k is an integer from 1 to 16, more particularly k is 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, or 16.
  • the lipid-like moiety Rz is -(C2-C2o)alkenyl, more particularly -(Ce-Cisjalkenyl, and even more particularly -(Ci2-Cis)alkenyl.
  • the lipid-like moiety Rz has the formula (XXIV): wherein k is an integer from 1 to 16, more particularly k is 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, or 16, and q is an integer from 0 to 18, more particularly q is 0, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, or 18.
  • the lipid-like moiety Rz has the formula (XXIV), wherein k is an integer from 10 to 16, and q is an integer from 0 to 18.
  • the lipid-like moiety Rz has the formula (XXV): wherein p is an integer from 1 to 6, more particularly k is 1, 2, 3, 4, 5, 6.
  • the lipid-like moiety Rz has the formula (XXVI): wherein k is an integer from 1 to 16, more particularly k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16, and p is an integer from 1 to 6. More particularly, the lipid-like moiety R7 has the formula (XXVI) wherein k is an integer from 12 to 16, and p is an integer from 1 to 6.
  • the lipid-like moiety Rz has the formula (IX), wherein g is an integer from 0 to 16, more particularly, g is from 0 to 12, and even more particularly g is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and each I is independently an integer from 0 to 18, more particularly, each I is independently from 0 to 12, and even more particularly, each I is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. Even more particularly, the lipid-like moiety Rz has the formula (IX), wherein g is an integer from 10 to 16; and each I is independently an integer from 0 to 18.
  • the lipid-like moiety Rz has the formula (X), wherein each g is independently an integer from 0 to 16, more particularly, each g is independently from 0 to 12, and even more particularly each g is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and I is an integer from 0 to 18, more particularly, I is from 0 to 12, and even more particularly, I is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. Even more particularly, the lipid-like moiety Rz has the formula (X), wherein each is independently from 8 to 12; and I is from 0 to 12
  • the lipid-like moiety Rz has the formula (XI), wherein I is an integer from 0 to 18, more particularly, I is from 0 to 12, and even more particularly, I is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; Y and Y' are independently selected from -OH, -COORx, and -OCORx; and each Rx is independently -(Ci-Ci8)alkyl or -(C2-Ci8)alkenyl.
  • the lipid-like moiety Rz has the formula (XI), wherein I is an integer from 0 to 18; Y and Y' are independently selected from -OH, -COORx, and -OCORx; and each Rx is independently -(Cio-Ci8)alkyl or -(Cio-Ci8)alkenyl.
  • each Rx is a radical independently selected from formula (xia) and (xib):
  • each Rx is a radical selected from formula (xia) and (xib) :
  • xia (xia) (xib) wherein the wavy bonds mean that the stereochemistry of the double bonds is undefined (i.e. it includes cis and trans isomers), k is an integer from 12 to 16, p is an integer from 12 to 18, q is an integer from 0 to 18; and r is an integer from 0 to 6.
  • Y and Y' are the same. In another particular embodiment, optionally in combination with any of the embodiments provided above or below, in the lipid-like moiety R?of formula (IX), Y and Y' are different.
  • the lipid-like moiety Rz of formula (XI) is selected from the formulas (xi1), (xi2), (xi3), (xi4), (xi5), (xi6), wherein each k is independently an integer from 1 to 16, more particularly each k is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; I is an integer from 0 to 18, more particularly, I is from 0 to 12, and even more particularly, I Is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; each p and each q are independently an integer from 0 to 18, more particularly each p and each q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18; and each r is independently an integer from 0 to 6, more particularly each r is independently 0, 1, 2, 3, 4, 5, or 6.
  • the lipid-like moiety Rz of formula (XI) is selected from the formulas (xi1), (xi2), (xi3), (xi4), (xi5), (xi6), (xi7) and (xi8), wherein each k is independently an integer from 12 to 16, 1 is an integer from 0 to 18, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • each k has the same meaning.
  • each p has the same meaning
  • each r has the same meaning
  • each q has the same meaning.
  • the lipid-like moiety Rz has the formula (XII), wherein Q and Q' are independently selected from -OCORy and -COORy; each Ry is independently -(Ci-Ci8)alkyl or -(C2-Ci8)alkenyl; each h is independently an integer from 1 to 18, more particularly, each h is independently from 1 to 12, and even more particularly each h is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and I is an integer from 0 to 18, more particularly, I is from 0 to 12, and even more particularly, I Is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rz has the formula (XII), wherein Q and Q' are independently selected from -OCORy and -COORy; each Ry is independently -(Cio-Ci8)alkyl or -(Cio-Ci8)alkenyl; each h is independently an integer from 1 to 18, and I is an integer from 0 to 18.
  • each Ry is a radical independently selected from formula (xiia), (xiib), and (xiic):
  • k is an integer from 1 to 16, more particularly k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16;
  • p and q are independently an integer from 0 to 18, more particularly p and q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18;
  • r is an integer from 0 to 6, more particularly r is 0, 1, 2, 3, 4, 5, or 6.
  • each Ry is a radical independently selected from formula (xiia), (xiib), and (xiic), wherein the wavy bonds mean that the stereochemistry of the double bonds is undefined (i.e. it includes cis and trans isomers), k is an integer from 12 to 16, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • the lipid-like moiety Rz of formula (XII) is selected from the formulas (xii1), (xii2), (xii3), and (xii4): wherein k is an integer from 1 to 16, more particularly k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; each p and each q are independently an integer from 0 to 18, more particularly each p and each q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18; each h is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rzof formula (XII) is selected from the formulas (xii 1 ), (xii2), (xii3), and (xii4), wherein k is an integer from 12 to 16, each h is independently an integer from 1 to 18, i is an integer from 0 to 18, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • the lipid-like moiety Rz has the formula (XIII), wherein Q and Q' are independently selected from -OCORy and -COORy; each Ry is independently -(Ci-Cis)alkyl or -(C2-Cis)alkenyl; each h is independently an integer from 1 to 18, more particularly, each h is independently from 1 to 12, and even more particularly each h is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rz has the formula (XIII), wherein Q and Q' are independently selected from -OCORy and -COORy; each Ry is independently -(Cio-Ci8)alkyl or -(Cio-Ci8)alkenyl; each h is independently an integer from 1 to 18, and i is an integer from 0 to 18. Even more particularly, in the lipid-like moiety Rz of formula (XIII), each Ry is a radical independently selected from formula (xiiia), (xiiib), and (xiiic):
  • k is an integer from 1 to 16, more particularly k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16;
  • p and q are independently an integer from 0 to 18, more particularly p and q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18;
  • r is an integer from 0 to 6, more particularly r is 0, 1, 2, 3, 4, 5, or 6.
  • each Ry is a radical independently selected from formula (xiiia), (xiiib), and (xiiic), wherein the wavy bonds mean that the stereochemistry of the double bonds is undefined (i.e. it includes cis and trans isomers), k is an integer from 12 to 16, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • Q and Q' are both -OCORy.
  • Q and Q' are both -COORy.
  • the lipid-like moiety Rz of formula (XIII) has the formula (xiiil):
  • each p and each q are independently an integer from 0 to 18, more particularly each p and each q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18;
  • r is an integer from 0 to 6, more particularly each r is independently 0, 1, 2, 3, 4, 5, or 6;
  • each h is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
  • i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rz of formula (XIII) is has the formula (xiiil), each h is independently an integer from 1 to 18, i is an integer from 0 to 18, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • the lipid-like moiety Rz has the formula (XIV), wherein i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and each j is independently an integer from 0 to 18, more particularly, each j is independently selected from 0 to 12, and even more particularly each j is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. More particularly, the lipid-like moiety Rz has the formula (XIV), wherein i is an integer from 0 to 18, and each j is independently an integer from 10 to 18.
  • the lipid-like moiety Rz has the formula (XV), wherein i is an integer from 0 to 18; more particularly, i is from 0 to 12, and even more particularly i is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and each j is independently an integer from 0 to 18, more particularly, each j is independently selected from 0 to 12, and even more particularly each j is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. More particularly, the lipid-like moiety Rz has the formula (XV), wherein i is an integer from 0 to 18, and each j is independently an integer from 10 to 18.
  • the lipid-like moiety Rz has the formula (XVI), wherein Q and Q' are independently selected from -OCORy and -COORy; Z is selected from -O-, -OCO-, -COO-, -NRz'CO-, and -CONRz'-; each Rz' is H or Rz; each Ry and Rz is independently -(Ci-Cis)alkyl or -(C2-Cis)alkenyl; and i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly i is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rz has the formula (XVI), wherein Q and Q' are independently selected from -OCORy and -COORy; Z is selected from -O-, -OCO-, -COO-, -NRz'CO-, and -CONRz'-; each Rz' is H or Rz; each Ry is independently -(Cio-Ci8)alkyl or -(Cio-Ci8)alkenyl; each Rz is independently -(Ci-Ci8)alkyl or -(C2-Ci8)alkenyl, and i is an integer from 0 to 18. Even more particularly, in the lipid-like moiety Rz of formula (XVI), each Ry is a radical independently selected from formula (xvia) and (xvib):
  • k is an integer from 1 to 16, more particularly k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16;
  • p and q are independently an integer from 0 to 18, more particularly p and q are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18;
  • r is an integer from 0 to 6, more particularly r is 0, 1, 2, 3, 4, 5, or 6.
  • each Ry is a radical independently selected from formula (xvia) and (xvib), wherein the wavy bonds mean that the stereochemistry of the double bonds is undefined (i.e. it includes cis and trans isomers), k is an integer from 12 to 16, p is an integer from 12 to 18, q is an integer from 0 to 18, and r is an integer from 0 to 6.
  • Q and Q' are both -OCORy.
  • Q and Q' are both -COORy.
  • Z is -O-.
  • Z is -OCO-.
  • Z is -CONRz’.
  • the lipid-like moiety Rz of formula (XVI) is selected from the formulas (xvi1), (xvi2), (xvi3), and (xvi4):
  • each k is independently an integer from 1 to 16, more particularly each k is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; and i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
  • the lipid-like moiety Rz of formula (XVI) is selected from the formulas (xvi1), (xvi2), (xvi3), and (xvi4), wherein each k is an integer from 12 to 16, and i is an integer from 0 to 18.
  • the lipid-like moiety Rz has the formula (XVII), wherein I is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is 0, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , or 12; and each j is independently an integer from 0 to 18, more particularly, each j is independently selected from 0 to 12, and even more particularly each j is independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , or 12. More particularly, the lipid-like moiety Rz has the formula (XVII), wherein i is an integer from 0 to 18, and each j is independently an integer from 10 to 18.
  • the lipid-like moiety Rz of formula (XVII) has the formula (xviil):
  • i is an integer from 0 to 18, more particularly, i is from 0 to 12, and even more particularly, i is 0, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , or 12.
  • the lipid-like moiety Rz has the formula (XVIII),
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a single bond
  • R22, R33 and R44 are selected from the following meanings: (i) R22, R33 and R44 are methyl, (ii) R22, R44 are methyl and R33 is hydrogen, (iii) R33 and R44 are methyl and R22 is hydrogen, and (iv) R44 is methyl and R22 and R33 are hydrogen.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a single bond and R22, R33 and R44 are methyl.
  • the formula (XVIII) has the formula (xviiil):
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a double bond
  • R22, R33 and R44 are selected from the following meanings: (I) R22, R33 and R44 are methyl, (II) R22, R44 are methyl and R33 is hydrogen, (ill) R33 and R44 are methyl and R22 is hydrogen, and (iv) R44 is methyl and R22 and R33 are hydrogen.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a double bond and R22, R33 and R44 are methyl.
  • the linker L is a biradical selected from the group consisting of -(Ci-Ci2)alkylene- optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -NR aa Rbb, -SH, -NHNH2, -OOOR C c, -CF3, and -OCF3, wherein R aa , Rbb and R cc are radicals independently selected from the group consisting of H, -phenyl, -(Ci-C6)alkyl, -(C2-C6)alkenyl, -(Ci-Cejalkylphenyl, and -phenyl(Ci-Ce)alkyl; -(Ci-Ce)alkyl-CO-; -CO-(Ci-Ce)alkyl-; and a biradical
  • L is a -(Ci-Ci2)alkylene-, more particularly a -(Ci-C6)alkyl-, and is optionally substituted as defined herein.
  • L is -(Ci-Ce)alkyl-CO- or -CO-(Ci-Ce)alkyl- and is optionally substituted as defined herein.
  • L is a biradical of formula (HID), more particularly a biradical of formula (HID) wherein b1 Is O, and a1 + c1 are selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12, and even more particularly, L is a biradical of formula (HID) selected from the group consisting of formula (IIIDa), formula (IIIDb), and formula (lllDc):
  • L is a biradical selected from the formulas (IVD), (VD), and (VID), more particularly, in the biradical of formula (IVD), a1 is 1 and the dashed bond is a double bond, i.e., the biradical of formula (IVD) has the formula (IVDa), and in the biradical of formula (VD), the dashed bond is more particularly a double bond, i.e., the biradical of formula (VD) has the formula (VDa).
  • L is a biradical selected from the formulas (VID), (VIID), and (VIIID), more particularly, in the biradical of formula (VIIID), a1 is 1.
  • L is a biradical of formula (IXD), more particularly a biradical of formula (IXD) wherein b1 and g1 are 0; and a1, d, d1, e1, f1 and hi are 1, i.e., a biradical of formula (IXDa).
  • L is a biradical selected from the formulas (XD) and (XID), more particularly in the biradicals of formula (XD) and (XID), b1 and g1 are 0; and a1, d, d1, e1, f1 and hi are 1, i.e. the biradicals of formula (XD), and (XID) have the formulas (XDa), and (XIDa), respectively.
  • L is a biradical selected from the formulas (XIID) and (XIIID), more particularly in the biradicals of formula (XIID) and (XIIID), b1 is O; and a1, d and d1 are 1.
  • L is a biradical selected from the formulas (XIVD), (XVD), (XVID), (XVIID), and (XVIIID), more particularly in the biradicals of formula (XIVD), (XVD), (XVID), (XVIID), and (XVIIID), b1 Is O; and a1, and d are 1.
  • L is a biradical of formula (XIXD), wherein p1 is 0, and j1 and k1 are 1.
  • L is a biradical of formula (XIXD), wherein p1 is 1, X1' is -NH-, and j1 and k1 are 1.
  • L is a biradical selected of formula (XIXD), wherein p1 is 1, X1' is -O-, and j1 and k1 are 1.
  • A is a lipid-like moiety Rz which has the formula (XVIII), t is 0, X is 0 and Ri is absent, i.e., the lipo-polyamino acid conjugate of formula (I) has the formula (Ih): wherein Ai, PAAi, PAA2, n, m, p, R22, R33, R44, and the dashed bonds — are as defined herein.
  • A is a lipid-like moiety Rz which has the formula (XVIII), wherein t is 1 , i.e., the lipo-polyamino acid conjugate of formula (I) has the formula (Ij): wherein Ai, PAAi, PAA2, X, R1, L, n, m, p, R22, R33, R44, and the dashed bonds — are as defined herein.
  • L is -CO-(Ci-C6)alkyl
  • X is N
  • R1 is H.
  • L is a biradical of formula (HID).
  • A is a lipid-like moiety Rz selected from the group consisting of: a) -(Ci-Ci8)alkyl, more particularly -(Ce-C jalkyl, and even more particularly, a radical of the formula (XXIII) as previously defined; b) -(C2-Ci8)alkenyl, more particularly -(Ce-C jalkenyl, and even more particularly, a radical of the formula (XXIV) as previously defined; c) a radical of formula (XXV) as defined herein; d) a radical of formula (XXVI) as defined herein; e) a radical of formula (IX) as defined herein; f) a radical of formula (X) as defined herein; g) a radical of formula (XI) as defined herein, more particularly, a radical selected from the formula
  • the lipo-polyamino acid conjugate of formula (I) has the formula (IC), wherein Ai, PAAi, PAA2, m, n, and p are as defined herein, and each k is independently an integer from 1 to 16, more particularly each k is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. Even more particularly, each k is independently an integer from 12 to 16. Even more particularly, in the lipo-polyamino acid conjugate of formula (IC), each k has the same meaning.
  • A is a lipid-like moiety Rz as defined herein, p is 0, and A1 is H.
  • A is a lipid-like moiety Rz as defined herein, p is 0, and A1 is an amino protective group.
  • Suitable amino protective groups known in the art may be used without limitation.
  • Non-limiting examples of amino protective groups include acyl-based groups, carbamate-based groups, imide-based groups, sulfonamide-based groups, and the like.
  • the amino protective group is selected from the group consisting of acetyl, methyloxycarbonyl, benzyloxycarbonyl (Cbz), p-methoxybenzyloxycarbonyl, t-butyloxycarbonyl (Boc), 9-fluorenyl methyloxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl group (Troc), benzoyl (Bz), benzyl (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), tosyl (Ts), trimethylsilylethoxycarbonyl (Teoc), benzhydryl, triphenylmethyl (Trityl), (4-methoxyphenyl)diphenylmethyl (MMT), dimethoxytrityl (DMT), and diphenylpho
  • A is a lipid-like moiety Rz as defined herein, p is 0, and Ai is an amino acid-like moiety Rs, being Rs a radical independently selected from the group consisting of (XX), (XXI) and (XXII)
  • each Rg is independently -(Ci-Cejalkyl, and Rw is selected from the group consisting of -(Ci-Cejalkyl-SOs- and -(Ci-Cejalkyl-COg-; and
  • A is a lipid-like moiety Rz as defined herein, p is 0, and Ai is selected from the group consisting of H, -(Ci-Cejalkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(Cs-C jaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, and -(C5-Cio)heterocycloalkyl; being Ai optionally substituted as defined herein, more particularly Ai is methyl.
  • A is a lipid-like moiety Rz as defined herein, p is 1 , Ai is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(Ce-C jaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, -(C1-C10) alkoxy, -(Ce-Cwjaryloxy, -(Ce-Cwjaralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alkyl-0-(C6-Cio)aryloxy; being Ai optionally substituted as defined herein, more
  • p is 1
  • A is a lipid-like moiety Rz
  • Ai is a lipid-like moiety Rz' as defined herein.
  • A is a lipid-like moiety Rz and Ai is a lipid-like moiety Rz' as defined herein.
  • A is a lipid-like moiety Rz as defined herein, p is 1
  • Ai is an active moiety selected from the group consisting of a pharmaceutically active agent, a cell-targeting agent, a penetration enhancing agent, a cosmetically active agent, and a diagnostically active agent.
  • the active moiety in Ai may be directly bound to the nitrogen atom of the lipo-polyamino acid conjugate or, alternatively, it may be bound through a linker.
  • a linker There is no limitation in the nature of the linker provided that it can be attached to the rest of the molecule by chemically feasible bonds.
  • the linker may contain from 1-50 moieties, more particularly from 2 to 30 or from 2 to 20 or from 2 to 15 moieties as defined above, and may be attached to the nitrogen atom of the PAA by a chemically feasible bond which is selected from the group consisting of an amine bond, an amide bond, a carbamate bond or a urea bond.
  • the lipid-like moiety Rz' in Ai in the lipo-polyamino acid conjugate of formula (I) is selected from the group consisting of -(Ci-C 2 o)alkyl, -(C 2 -C 2 o)alkenyl, and a radical of formula (IX), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII’).
  • the lipid- like moiety Rz' is -(Ci-C 2 o)alkyl, more particularly -(Ce-C jalkyl. More particularly, the lipid-like moiety Rz' has the formula (XXIII) as defined herein.
  • the lipid-like moiety Rz' is -(C 2 -C 2 o)alkenyl, more particularly -(Ce-Cisjalkenyl. Even more particularly, the lipid-like moiety Rz' has the formula (XXIV) as defined herein.
  • the lipid-like moiety Rz' has the formula (XXV) as defined herein.
  • the lipid-like moiety Rz' has the formula (XXVI) as defined herein.
  • the lipid-like moiety Rz' has the formula (IX) as defined herein.
  • the lipid-like moiety Rz' has the formula (X) as defined herein.
  • the lipid-like moiety R has the formula (XI) as defined herein.
  • Y and Y' are the same. In another particular embodiment, optionally in combination with any of the embodiments provided above or below, in the lipid-like moiety R of formula (XI), Y and Y' are different.
  • the lipid-like moiety R of formula (XI) is selected from the formulas (xi 1 ), (xi2), (xi3), (xi4), (xi5), (xi6), (xi7) and (xi8) as defined herein.
  • each k has the same meaning.
  • each p has the same meaning
  • each r has the same meaning
  • each q has the same meaning
  • the lipid- like moiety R has the formula (XII) as defined herein.
  • Q and Q' are both -OCORy.
  • Q and Q' are both -COORy.
  • the lipid-like moiety R of formula (XII) is selected from the formulas (xii1), (xii2), (xii3), and (xii4) as defined herein.
  • the lipid-like moiety R has the formula (XIII) as defined herein.
  • Q and Q' are both -OCORy.
  • Q and Q' are both -COORy.
  • the lipid-like moiety R of formula (XIII) has the formula (xiiil) as defined herein.
  • the lipid- like moiety R has the formula (XIV) as defined herein. In one embodiment, optionally in combination with any of the embodiments provided above or below, the lipid- like moiety R has the formula (XV) as defined herein.
  • the lipid-like moiety R has the formula (XVI) as defined herein.
  • Q and Q' are both -OCORy.
  • Q and Q' are both -COORy.
  • Z' is -O-.
  • Z' is -OCO-.
  • Z' is -CONRz'.
  • the lipid-like moiety R of formula (XVI) is selected from the formulas (xvi1), (xvi2), (xvi3), and (xvi4) as defined herein.
  • the lipid-like moiety R has the formula (XVII) as defined herein. In one particular embodiment, optionally in combination with any of the embodiments provided above or below, the lipid-like moiety R of formula (XVII) has the formula (xviil) as defined herein.
  • the lipid-like moiety R has the formula (XVIII'), wherein t', R22, R33, R44, the dashed bonds — , and L are as defined herein.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a single bond
  • R22, R33 and R44 are selected from the following meanings: (i) R22, R33 and R44 are methyl, (ii) R22, R44 are methyl and R33 is hydrogen, (iii) R33 and R44 are methyl and R22 is hydrogen, and (iv) R44 is methyl and R22 and R33 are hydrogen.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a single bond and R22, R33 and R44 are methyl.
  • the formula (XVIII') has the formula (xviiil '): wherein t', the dashed bonds — , and L are as defined herein.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a double bond
  • R22, R33 and R44 are selected from the following meanings: (I) R22, R33 and R44 are methyl, (II) R22, R44 are methyl and R33 is hydrogen, (ill) R33 and R44 are methyl and R22 is hydrogen, and (iv) R44 is methyl and R22 and R33 are hydrogen.
  • each of the dashed bonds of the alkyl chain attached to the chroman ring is a double bond and R22, R33 and R44 are methyl.
  • the linker L is a biradical selected from the group consisting of -(Ci-Ci2)alkylene- optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -NR aa Rbb, -SH, -NHNH2, -OOOR C c, -CF3, and -OCF3, wherein R aa , Rbb and R cc are radicals independently selected from the group consisting of H, -phenyl, -(Ci-C6)alkyl, -(C2-C6)alkenyl, -(Ci-Cejalkylphenyl, and -phenyl(Ci-C6)alkyl; -(Ci-C6)alkyl-CO-; and a biradical of formula (HID), (IVD), (VD), (VID
  • L is a -(Ci-Ci2)alkylene-, more particularly a -(Ci-Cejalkyl-, and is optionally substituted as defined herein.
  • L is -(Ci-Ce)alkyl-CO- or -(Ci-Ce)alkyl- and is optionally substituted as defined herein.
  • L is a biradical of formula (HID)
  • L is a biradical of formula (HID), more particularly a biradical of formula (HID) wherein b1 is 0, and a1 + c1 are selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , and 12, and even more particularly, L is a biradical of formula (HID) selected from the group consisting of formula (IIIDa), formula (IIIDb), and formula (IHDc):
  • L is a biradical selected from the formulas formulas (IVD), (VD), and (VID), more particularly, in the biradical of formula (IVD), a1 is 1 and the dashed bond is a double bond, i.e., the biradical of formula (
  • L is a biradical selected from the formulas (VID), (VIID), and (VIIID) as defined herein, more particularly, in the biradical of formula (VIIID), a1 is 1.
  • L is a biradical of formula (IXD), more particularly a biradical of formula (IXD) wherein b1 and g1 are 0; a1, d, d1, e1, f1 and hi are 1, i.e. a biradical of formula (IXDa) as defined herein.
  • L is a biradical selected from the formulas (XD') and (XID'), more particularly in the biradicals of formula (XD) and (XID), b1 and g1 are 0; and a1, d, d1, e1, f1 and hi are 1, i.e. the biradicals of formula (XD), and (XID) have the formulas (XDa), and (XIDa), respectively, as defined herein.
  • L is a biradical selected from the formulas (XIID) and (XIIID), more particularly in the biradicals of formula (XIID) and (XIIID) as defined herein, b1 is 0; a1, d and d1 are 1.
  • L is a biradical selected from the formulas (XIVD), (XVD), (XVID), (XVIID), and (XVIIID), as defined herein, more particularly in the biradicals of formula (XIVD), (XVD), (XVID), (XVIID), and (XVIIID), b1 is O; and a1, and d are 1.
  • L is a biradical selected from the formulas (XIXD), wherein p1 is 0, and j1 and k1 are 1.
  • L is a biradical of formula (XIXD) as defined herein, wherein p1 is 1, X1' is -NH-, and j1 and k1 are 1.
  • L is a biradical selected of formula (XIXD), wherein p1 is 1 , X1’ is -O-, and j1 and k1 are 1.
  • Ai is a lipid-like moiety R which has the formula (XVIII'), p is 0, s is 0, and t' is 1 , i.e., the lipo-polyamino acid conjugate of formula (I) has the formula (Ih'): wherein A, X, Ri, PAAi, PAA2, n, m, R22, R33, R44, L, and the dashed bonds — are as defined herein.
  • A1 is a lipid-like moiety R which has the formula (XVIII'), wherein p is 1 , i.e., the lipo-polyamino acid conjugate of formula (I) has the formula (lj'): wherein A, X, R1, PAA1, PAA2, n, m, R22, R33, R44, L, t', and the dashed bonds — are as defined herein
  • L is -(Ci-Ce)alkyl-CO- or a biradical of formula (HID).
  • the lipid-like moiety R has the formula (XVIII”):
  • lipid-like moiety R has the formula (XVIII”), wherein b1 is 0, and a1 and d are independently selected from 2 to 6.
  • A1 is a lipid-like moiety R selected from the group consisting of: a) -(Ci-Ci8)alkyl, more particularly -(Ce-Cisjalkyl, and even more particularly, a radical of the formula (XXIII) as previously defined; b) -(C2-Ci8)alkenyl, more particularly -(Ce-Cisjalkenyl, and even more particularly, a radical of the formula (XXIV) as previously defined; c) a radical of formula (XXV) as defined herein; d) a radical of formula (XXVI) as defined herein; e) a radical of formula (IX) as defined herein; f) a radical of formula (X) as defined herein; g) a radical of formula (XI) as defined herein, more particularly, a radical selected from
  • A1 is a lipid-like moiety R as defined herein, p is 1, and A is H.
  • A1 is a lipid-like moiety R as defined herein, p is 1, and A is a protective group.
  • Ai is a lipid-like moiety R as defined herein, p is 1, X is N, and A is an amino protective group; more particularly the amino protective group is selected from the group consisting of acetyl, methyloxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, t-butyloxycarbonyl (Boc), 9- fluorenylmethyloxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2,2,2-trichloro-ethoxycarbonyl group (Troc), benzoyl (Bz), benzyl (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), tosyl (Ts), trimethylsilylethoxy-carbonyl (Teoc),
  • Ai is a lipid-like moiety R as defined herein, p is 1, X is 0, and A is a carboxy protective group.
  • Suitable carboxy protective groups known in the art may be used without limitation.
  • Representative carboxy protective groups include alkyl, aryl or benzyl esters, silyl esters, amides or hydrazides.
  • the carboxy protective group is selected from the group consisting of -(Ci-Cejalkyl, benzyl, p-methoxyphenyl, trimethylsilyl and [2-(trimethylsilyl)ethoxy]methyl (SEM).
  • Ai is a lipid-like moiety R as defined herein, p is 1, X is S, and A is selected from the group consisting of -(Ci-Cejalkyl, benzyl, p-methoxyphenyl, trimethylsilyl and [2-(trimethylsilyl)ethoxy] methyl (SEM).
  • Ai is a lipid-like moiety R as defined herein, p is 1, and A is selected from the group consisting of H, -(Ci-Cejalkyl, -CO(Ci-C6)alkyl, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(Ce-C jaralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, a radical (IV), a radical (V), a radical (VI), a protective group, a lipid-like moiety Rz, an active moiety; being A optionally
  • Ai is a lipid-like moiety Rz' as defined herein, p is 1, and A is an active moiety selected from the group consisting of a pharmaceutically active agent, a cell-targeting agent, a penetration enhancing agent, a cosmetically active agent, and a diagnostically active agent.
  • the active moiety in A may be directly bound to X or, alternatively, it may be bound through a linker.
  • the linker may contain from 1-50 moieties, more particularly from 2 to 30 or from 2 to 20 or from 2 to 15 moieties as defined above and may be attached to X by a chemically feasible bond such as an ether bond, a thioether bond, or an amide bond.
  • PAA1 is a repetitive unit of formula (II”) wherein though the repeating units PAA1” and PAA2” are shown in a particular order for convenience of description, the repeating units may be present in any order and may be block or randomly present; and wherein each of the repeating units PAA1” and PAA2” may comprise blocks of monomer units which may be the same or different between each other; n' is an integer from 5 to 250; m' is an integer from 5 to 250;
  • Xi is selected from the group consisting of CH, N, S, and 0;
  • Ria is a radical which has any of the meanings of R1, with the condition that Ri a is absent when Xi is 0 or S;
  • R 2 ' is a radical which has any of the meanings of R 2 ;
  • R3' is a radical which has any of the meanings of R3;
  • R4' is a radical which has any of the meanings of R4;
  • R5' is a radical which has any of the meanings of R5;
  • B and B’ are independently S or CH 2 ;
  • e and f are independently an integer from 1 to 18;
  • A3 is selected from the group consisting of H, -(Ci-C6)alkyl, -CO(Ci-C6)alkyl, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, a radical (IV), a radical (V), a radical (VI), a protective group, a lipid-like moiety Rz, and an active moiety; wherein is optionally substituted by one or more groups selected from the group consisting of -OH, halogen, -CF3, -NH2, -NH-(Ci-C4)alkyl, -NH-CO-(Ci- C 6
  • the lipo-polyamino acid conjugate of formula (ID) is a dimer wherein A and A3 have the same meaning, X and Xi have the same meaning, R1 and Ri a have the same meaning, R 2 and R 2 ' have the same meaning, R3 and R3' have the same meaning, R4 and R4’ have the same meaning, R5 and R5’ have the same meaning, B and B’ have the same meaning, e and f have the same meaning, n and n' have the same meaning, and m and m' have the same meaning, respectively.
  • PAA/ is a repetitive unit of formula (II')
  • PAA2' is a repetitive unit of formula (III') wherein R2', R3', R4', and R5' are as defined herein; p' is 0 or 1 ; s' is 0 or 1 ;
  • X is selected from the group consisting of N, and 0;
  • Ri is a radical which has any of the meanings of R1', with the condition that R is absent when X” is 0;
  • A2 is selected from the group consisting of H, -OH, -(Ci-Cejalkyl,
  • A2 is selected from the group consisting of H, -(Ci-C6)alkyl, -(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, -(Ci-Cio)alkoxy, -(C6-Cio)aryloxy, -(C6-Cio)aralkoxy, -(C5-Cio)heteroaralkoxy, -(Ci-Cio)alkyl-0-(C6-Cio)aryloxy, an amino protective group, a lipid-like moiety R , an amino acid-like moiety Rs, and an active moiety; with the condition that: when p' is 0 and s' is 0, A2 is selected from the group consisting of H, -(Ci-C6)alkyl, -(C5-Cio)aryl, -(C5-Cio)heteroaryl, -(C6-Ci
  • the lipo-polyamino acid conjugate of formula (ID') is a dimer wherein A1 and A2 have the same meaning, X and Xi have the same meaning, X' and X” have the same meaning, R/ and R1” have the same meaning, R1 and Ri a have the same meaning, R2 and R2' have the same meaning, Rs and R3' have the same meaning, R4 and R4' have the same meaning, Rs and R5' have the same meaning, B and B’ have the same meaning, e and f have the same meaning, n and n' have the same meaning, and m and m' have the same meaning, p and p' have the same meaning, and s and s' have the same meaning, respectively.
  • the lipo-polyamino acid conjugate of formula (I) has the formula (IA) wherein: n is an integer from 5 to 100; X is N; R1 is selected from the group consisting of H, and -(Ci-Ce)alkyl;
  • R2 is a radical selected from the group consisting of (IV) and (VI); R3 is H; p is 0 or 1 ; s is 0 or 1 ; X' is N;
  • R1' is selected from the group consisting of H, and -(Ci-Ce)alkyl;
  • A is H, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, or a lipid-like moiety Rz which is selected from the group consisting of H, -(Ci-C2o)alkyl, -(C2-C2o)alkenyl, and a radical of formula (XI), (XII), (XIV), (XV), (XVI), (XVII) or (XVIII); and
  • A1 is H or a lipid-like moiety Rz' which is selected from the group consisting of -(Ci-C2o)alkyl, -(C2-C2o)alkenyl, and a radical of formula (XI), (XII), (XIV), (XV), (XVI), (XVII) or (XVIII').
  • the lipo-polyamino acid conjugate of formula (I) has the formula (IB) (IB) wherein: n is an integer from 5 to 100; m is an integer from 5 to 100; X is N; Ri is selected from the group consisting of H, and -(Ci-Ce)alkyl; R2 is a radical selected from the group consisting of (IV) and (VI); R3 is H; R4 corresponds to the amino acid the side chain of phenylalanine, alanine, leucine, glycine, isoleucine, or valine; R5 is H; p is 0 or 1 ; s is 0 or 1 ; X' is N; R/ is selected from the group consisting of H, and -(Ci-Cejalkyl; A is H, -(Ci-Ce)alkyl- CO-N[(Ci-Ci8)alkyl]
  • the lipid-like moiety Rz is -(Ci-C2o)alkyl, a radical of formula (XI), or a radical of formula (XVIII); and the lipid-like moiety Rz’ is -(Ci-C2o)alkyl, a radical of formula (XI), or a radical of formula (XVIII').
  • t is 1
  • L is -CO-(Ci-C6)alkyl, or a biradical of formula (HID)
  • t is 1
  • L is -(Ci-CeJalkyl-CO-, or a biradical of formula (HID).
  • the lipo-polyamino acid conjugate of formula (I) has the formula (IB'): wherein:
  • X is N or CH
  • A is selected from the group consisting of butyl, (Ci2-Ci8)alkyl, a radical of formula (XXV), and a radical of formula (XXVI);
  • R1 is selected from the group consisting of H and (Ci-Cis)alkyl
  • R is selected from the group consisting of H, -CO(Ci-C6)alkyl, a radical of formula (XVIII”), and -CONH(Ci-Ci8)alkyl;
  • R2 is as defined herein, particularly R2 is a radical of formula (VI) or a radical of formula (VI), wherein more particularly in the radical of formula (VI), c is an integer from 0 to 3;
  • R is -(Ci-C6)alkyl, and R' is -(Ci-Ce)alkyl- CO2-.
  • R4 is selected from the group consisting of H, methyl; isopropyl, isobutyl, and sec-butyl; n is a value from 5 to 200, or from 5 to 150, or from 10 to 150, particularly measured by 1 H-NMR spectroscopy; and m is 0 or a value from 5 to 150, or from 5 to 100, or from 5 to 80, or from 10 to 80, particularly measured by 1 H-NMR spectroscopy.
  • the lipo-polyamino acid conjugates of formula (I) must contain at least one lipid-like moiety, which may be either a lipid-like moiety Rz in A, a lipid-like moiety R/ in A1, or both.
  • the process for the preparation of the lipo-polyamino acid conjugates of formula (I) generally comprises polymerizing N-carboxy anhydrides (NCA) of protected or non-protected amino acids, to produce a poly(amino acid), or protected poly amino acid ester, carbamate, S-alkylsulfonyl, or trifluoroacetyl derivative. Then a deprotection step or thiol exchange should be carried out by methods well known for a person skilled in the art. The different radicals present in the repeating units may be introduced at desired ratios by changing the ratios of the respective block or random copolymers.
  • NCA N-carboxy anhydrides
  • the process comprises: i) reacting an amine or tetrafluoroborate or trifluoroacetate ammonium salt form of an initiator i.1) with an appropriate N-carboxyanhydride (NCA); alternatively, reacting the amine or tetrafluoroborate or trifluoroacetate ammonium salt form of initiator of step i) with an appropriate N- carboxyanhydrides in a sequential manner to obtain a block co-polymer; or alternatively, i.2) reacting the amine or tetrafluoroborate or trifluoroacetate ammonium salt form of initiator of step i) with an appropriate NCA mixture in a statistical manner to obtain random co-polymers; ii) optionally, reacting the amine group at the N-terminal position with an amine reactive group to introduce R1; iii) optionally, orthogonally removing amino acid side chain protecting groups;
  • Step I) above may include: a) ring opening polymerization of amino acids N-carboxyanydride (NCA) monomer by reacting the amine or tetrafluoroborate or trifluoroacetate ammonium salt form of initiator with the selected NCA, wherein the ratio of monomer/initiator allows the control of the degree of polymerization (DP); b) a sequential polymerization, wherein block co-polypeptides are prepared following the polymerization reaction a') in a sequential manner, allowing the first NCA monomer to be consumed and the resulting product may be purified or not before adding the next monomer to build the following polypeptidic block; or c) a statistical polymerization a') wherein random copolypeptides are prepared following the polymerization reactionin a statistical manner, mixing all the NCA monomers before starting the polymerization by the addition of an amine or tetrafluoroborate or trifluoroacetate ammonium salt form of initiator.
  • Step II) above corresponds to the end-capping, wherein the amine group at the N-terminal position is reacted with an amine reactive group to introduce Ri.
  • Step iii) above corresponds to the exchange reaction or deprotection, wherein amino acid side chains are removed orthogonally depending on the protecting group.
  • Step iv) corresponds to the conjugation, reacting the amine or carboxylic acid group at side chain terminal position to achieve a shielding moiety, by chloracetylation, methylation, thiol exchange, nucleophilic substitution or peptide coupling reactions in a sequential manner if needed.
  • the lipo-polyamino acid conjugates of the invention have amphiphilic nature since they comprise a polyamino acid-based moiety (hydrophilic part) and at least one lipid-like moiety (hydrophobic part). Thanks to their amphiphilic nature, the lipo-polyamino acid conjugates of the invention may form selfassembled particles in solution.
  • the present invention also relates to a self-assembled particle comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, and optionally one or more active agents selected from the group consisting of pharmaceutically active agents, penetration enhancing agents, cell-targeting agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • the present invention relates to a self-assembled particle comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, and one or more nucleic acids.
  • self-assembled particles refers to the arrangement of the lipo-polyamino acid conjugate molecules in a solvent, in particular, small, self-assembled particle may be enclosed structures of any shape, typically spherical and/or tubular.
  • the term “self-assembled particles” intends to encompass any of a number of structures that are known in the art to be formed from amphiphilic polymers.
  • Non-limiting examples of self-assembled particles include micelles (also being referred to interchangeably herein as micellar worms or simply “worms”), inverted micelles, planar bilayers, crystal nanoparticles, liposomes, microbubbles or lipid nanoparticles.
  • the self-assembled nanoparticles and microparticles can also form gels.
  • the self-assembled particles of the invention are non-viral particles, which means that they are not able to virally infect cells.
  • the self-assembled particles may show a variety of sizes, in particular, they can be nanoparticles or microparticles.
  • a “nanoparticle”, as defined herein, is any particle of nanometric size, in particular having smallest end-to-end diameter of between 1 and 900, more particularly, between 1 and 700 nm, between 1 and 500 nm, between 1 and 300 nm, between 1 and 200 nm, and between 1 and 100 nm in size.
  • a "microparticle”, as defined herein, is typically any particle of micrometric size, having a smallest end-to-end between 1 and 100 m in size.
  • the relevant diameter is the number average diameter.
  • the term "size" refers to a characteristic physical dimension.
  • the size of the particle corresponds to the diameter of the particle.
  • the size of the particle is determined by the diameter of the two relevant cross-section dimensions of the particle.
  • the size of the nanoparticle corresponds to the maximum edge length.
  • the set of particles can have a distribution of sizes around the specified size.
  • a size of a set of particles can refer to a mode of a distribution of sizes, such as a peak size of the distribution of sizes.
  • the term "diameter” refers to the average diameter and is also designated as Z-average or Z- ave.
  • the average diameter corresponds to the mean hydrodynamic diameter (Dh) and can be measured by dynamic light scattering (DLS) as shown in the examples below.
  • the self-assembled particles of the invention have a hydrodynamic diameter (Dh) in water from 5 to 1200 nm, more particularly, from 10 to 1100 nm, from 10 to 1000 nm, from 10 to 700, from 20 to 500 nm, from 20 to 400 nm, from 20 to 300 nm, from 30 to 200 nm, or from 50 to 150 nm.
  • the lipid-like moieties face inward and the polar head portion (the PAA) face outward.
  • the self-assembled particle is a core-shell structure comprising an inner core and an external shell. More particularly, the lipo-polyamino acid conjugate of formula (I) forms the external shell of the self-assembled particle, and an agent is optionally encapsulated or loaded in the inner core.
  • the self-assembled particle is a nanoparticle, more particularly selected from the group consisting of a micelle, an inverted micelle, a planar bilayer, a crystal nanoparticle, a liposome, microbubbles, and a lipid nanoparticle, and even more particularly, the self-assembled particle is a lipid nanoparticle or a liposome.
  • liposome refers to artificially prepared self-assembled particles composed of concentric lipidic bi-layers enclosing one or more aqueous compartments.
  • Non-limiting examples of liposomes include monolamellar liposome, multi-lamellar liposomes, multi-vesicular-liposomes and polymer-coated liposomes.
  • lipid nanoparticle refers to particles typically spherical on the order of nanometers (e.g., 1-1000 nm) that includes lipids and that is stable and dispersible in aqueous media.
  • crystal nanoparticle refers to artificially prepared crystals composed of solid lipids enclosing one or more hydrophobic compartments.
  • crystal nanoparticle refers to particles of different shapes on the order of nanometers (e.g., 1-1000 nm) that includes lipids and that is stable and dispersible in aqueous media.
  • the invention in another embodiment, optionally in combination with one or more features of the various embodiments described above or below, relates to a lipid nanoparticle (LNP) or a liposome comprising the lipo-polyamino acid conjugate of formula (I) as defined herein in the first or the second aspect, and optionally one or more active agents selected from the group consisting of pharmaceutically active agents, cell-targeting agents, penetration enhancing agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • active agents selected from the group consisting of pharmaceutically active agents, cell-targeting agents, penetration enhancing agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • the liposomes and lipid nanoparticles disclosed herein may comprise any suitable lipids, including ionizable lipids, cationic lipids, zwitterionic lipids, neutral lipids, or anionic lipids.
  • Suitable ionizable lipids include, without limitation, (2S)-2,5- bis(3-aminopropylamino)- N-[2- (dioctadecylamino)acetyl]pentanamide (DOGS), N1-[2-((1 S)-1-[(3-aminopropyl)amino]-4-[di(3-aminopropyl)- amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide (MVL5), 3
  • Suitable cationic lipids include, without limitation, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N- distearyl-N,N-dimethylammonium bromide (DDA13), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(1-(2,3-dioleyloxy)-propyl)-N,N,N-trimethylammonium chloride (DOTMA), N-[1-(2,3,- ditetradecyloxy)propyll-N,N-dimethyl-N-hydroxyethylammonium bromide (DMRIE), N-[1-(2,3,dioleyloxy)- propyl]-N, N-dimethyl-N-hydroxy ethylammonium bromide (DORIE), 3
  • anionic lipids include, but are not limited to, phosphatidylglycerol, diacylphosphatidylserine, diacylphosphatidic acid, N-Succinyl phosphatidylethanolamine, N-glutaryl phosphatidylethanolamine cholesterol hemisuccinate (CHEMS), lysylphos-phatidylglycerol, N-dodecanoyl phosphatidyl ethanoloamine, cardiolipin, and combinations thereof.
  • Suitable neutral lipids may be uncharged or zwitterionic lipids and include, without limitation steroids, phospholipids, and combinations thereof.
  • steroids examples include, without limitation, cholesterol, progesterone, cortisone, aldosterone, estradiol, testosterone, and combinations thereof.
  • phospholipids include, but are not limited to, phosphatidylcholine (PC), phosphatidic acid (PA), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylserine (PS), and phosphatidylinositol (PI), dimyristoyl phosphatidyl choline (DMPC), distearoyl phosphatidyl choline (DSPC), dioleoyl phosphatidyl choline (DOPC), dipalmitoyl phosphatidyl choline (DPPC), dimyristoyl phosphatidyl glycerol (DMPG), distearoyl phosphatidyl glycerol (DSPG), dioleoyl phosphatidyl glycerol (DOPG), dipalmitoyl phosphatidyl glycerol (DPPG), dimyristoyl phosphatidyl serine (DMDM
  • the lipo-polyamino acid conjugate advantageously provides stealth properties.
  • stealth refers to the fact that the liposomes or lipid nanoparticles are not detected and sequestered and/or degraded by the immune system of the host to which they are administered, and consequently the length of time for which the liposomes or lipid nanoparticles can exist in vivo is increased.
  • the self-assembled particles of the invention in particular liposomes and lipid nanoparticles, lack polyethyleneglycol (PEG).
  • PEG polyethyleneglycol
  • the liposomes and lipid nanoparticles disclosed herein may also comprise one or more cationic polymers such as cationic amine-containing polymers.
  • these polymers may carry active ingredients, particularly nucleic acids.
  • cationic amine-containing polymers include, without limitation, poly-L- lysine, polyamidoamine, polyethyleneimine, chitosan, poly(beta-amino esters), and the like.
  • the invention in another embodiment, optionally in combination with one or more features of the various embodiments described above or below, relates to a lipid nanoparticle (LNP) or a liposome comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, and one or more lipids selected from the group consisting of ionisable lipids, cationic lipids, neutral lipids, and anionic lipids. More particularly, the lipid nanoparticle (LNP) or liposome comprises the lipo-polyamino acid conjugate of formula (I) as defined herein, a ionizable lipid or a cationic lipid, a phospholipid, and a sterol.
  • the lipid nanoparticle (LNP) or liposome comprises: i) the lipo-polyamino acid conjugate of formula (I) as defined herein in an amount from 0.1 to 10 mol%, more particularly from 1 to 5 mol% or from 1 to 6 mol% or from 0.5 to 5 mol%, even more particularly from 2 to 5% mol%; ii) a ionizable lipid or a cationic lipid in an amount from 30 to 70 mol%, more particularly from 40 to 60 mol%; iii) a phospholipid in an amount from 1 to 20 mol%, more particularly from 5 to 15 mol%; and iv) a sterol in an amount from 20 to 60 mol%, more particularly from 30 to 50 mol%; wherein the percentages are expressed with respect to the sum of the mol% of the lipids and the lipo-polyamino acid conjugate of formula (I).
  • the invention in another embodiment, optionally in combination with one or more features of the various embodiments described above or below, relates to a lipid nanoparticle (LNP) or a liposome comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, one or more lipids selected from the group consisting of ionisable lipids, cationic lipids, neutral lipids, and anionic lipids, and one or more nucleic acids.
  • LNP lipid nanoparticle
  • a liposome comprising the lipo-polyamino acid conjugate of formula (I) as defined herein, one or more lipids selected from the group consisting of ionisable lipids, cationic lipids, neutral lipids, and anionic lipids, and one or more nucleic acids.
  • the lipid nanoparticles and the liposomes containing one or more nucleic acids may be prepared by standard methods.
  • the process for the preparation of LNPs comprises: i) preparing a first alcoholic mixture comprising one or more lipids and the lipo-polyamino acid conjugate of the invention in a suitable alcohol such as for example ethanol; ii) preparing a second aqueous composition comprising one or more nucleic acids and an aqueous solvent (an acidic buffer); and iii) mixing i) with ii) in a microfluidic mixing device.
  • the microfluidic mixing allows thorough and rapid mixing of the lipid phase and the nucleic acid phase in a microscale device.
  • the skilled person will be able to modulate the size of the LNPs.
  • the lipo-polyamino acid conjugate of the invention may contain one or more active moieties in its structure in any of A, Ai, A2 or A3.
  • active moieties are independently selected from the group consisting of a pharmaceutically active agent, a cell-targeting agent, a penetration enhancing agent, a cosmetically active agent, and a diagnostically active agent.
  • the lipo-polyamino acid conjugate of the invention may form self-assembled particles which may comprise one or more active agents selected from the group consisting of pharmaceutically active agents, cell-targeting agents, penetration enhancing agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • the term “pharmaceutically active agent” refers to and agent that has pharmacological activity and is used for curing, mitigating, treating or preventing a disease in a mammal, in particular a human.
  • pharmaceutical active agent refers to an agent that does not provide any therapy but is used for aesthetic purposes, for example to improve the appearance, preserve, condition, cleanse, color or protect the skin, nails or hair.
  • the term "cell-targeting agent” refers to any molecule, macromolecule, or biomacromolecule, displaying affinity for a molecule present in the human or animal body, which is able to direct the conjugates or the self-assembled particles thereof by directing them towards the target site for therapeutic treatment since e.g., it selectively binds to receptors that are expressed or over-expressed on specific cell types.
  • Celltargeting groups are well known in the art. The term therefore includes ligands for specific receptors or antigens, such as antibodies for a specific antigen, folic acid for its receptor or sugars such as galactose for its hepatic receptors.
  • the targeting agent may be attached to the lipo-polyamino acid conjugate backbone in any of A, Ai, A2 or A3 and/or may be contained in the self-assembled particles formed from the conjugated.
  • cell-targeting groups include, but are not limited to, galactosamine, folate, a Her-2 binding peptide, TLR agonists, p-D-Glucose, Asn-Gly-Arg peptide, angiopep2, folic acid, aptamers (A-9, A10, Anti- gp120, TTA1, sgc8, Anti MUC-1, AS1411), primaquine, zidovudine, superoxide dismutase, prednisolone, platinum, cisplatin, sulphamethoxazole, amoxicillin, etoposide, mesalzine, doxorubicin, paclitaxel, 5-amino salicylic acid, denosumab, docetaxel, calcitonin, proanthocyanidin, methotrexate, camptothecin, galactose, glycyrrhetinic acid, lactose, hyaluor
  • RCPLSHSLICY laminin receptor binding peptide
  • laminin receptor binding peptide e.g. YIGSR
  • a skin homing peptide a retina homing peptide, a pancreas homing peptide, a liver homing peptide, a lymph node homing peptide, an adrenal gland homing peptide, a thyroid homing peptide, a bladder homing peptide, a breast homing peptide, a neuroblastoma homing peptide, a lymphoma homing peptide, a muscle homing peptide, a wound vasculature homing peptide, an adipose tissue homing peptide, a virus binding peptide, or a fusogenic peptide.
  • YIGSR laminin receptor binding peptide
  • penetration enhancing agent refers to moiety or compound that increases the permeability of an active agent, in particular selected from a pharmaceutically active agent, a cell-targeting agent, a cosmetically active agent, and a diagnostically active agent.
  • the penetration enhancing agent is also known as permeation enhancer.
  • penetration enhancing agents include, without limitation, cell penetrating peptides, surfactants, terpenes, sulfoxides, pyrrolidones, fatty acids, fatty alcohols, urea, azones, fatty alcohols, fatty acids, fatty esters; such as for example, lauryl sarcosine, octoxynol, phenylsulfonate, pluronic, sodium laurate, sodium oleate, sorbitan dilaurate, sorbitan dioleate, sorbitan trilaurate, sorbitan trioleate, sodium octyl sulfate, alkyl ammonium halides, decanol, dodecanol, linolenyl alcohol, oleyl alcohol, butyl acetate, cetyl lactate, lauryl lactate, myristyl lactate, diethyl sebacate, diethyl succinate, di
  • pharmaceutically active agents include low molecular weight drugs, peptides, antibodies, hormones, enzymes, nucleic acids, proteins, and combinations thereof.
  • nucleic acid refers to DNA or RNA.
  • the nucleic acid is an DNA/RNA hybrid, a short interfering RNA (siRNA), a microRNA (miRNA), a single guide RNA (sgRNA), a donorDNA, a self- amplyfing/replicating RNA, a circularRNA (oRNA), a plasmid DNA (pDNA), a closed-linear DNA (clDNA), a short hairpin RNA (shRNA), messenger RNA (mRNA), and antisense RNA (aRNA), a messenger RNA (mRNA), a CRISPR guide RNA, an antisense nucleic acid, a decoy nucleic acid, an aptamer, and a ribozyme to name a few, and encompasses both the nucleotide sequence and any structural embodiments thereof, such as double stranded, single strand
  • the nucleic acid may be a poly- or oligonucleotide, such as oligo- or poly-double stranded RNA, oligo- or polydouble stranded DNA, oligo- or poly-single stranded RNA, oligo- or poly-single stranded DNA.
  • Each of the nucleotides contained in the nucleic acid may be a naturally occurring nucleotide or a chemically-modified, non-naturally occurring nucleotide.
  • the strand length of the nucleic acid is not particularly limited and the nucleic acid may have a short strand ranging from 10 to 200 bases, preferably from 20 to 180 bases, preferably from 25 to 100 bases, preferably from 30 to 50 bases; or the nucleic acid may have a relatively long strand of from 200 to 20000 bases, more preferably of from 250 to about 15000 bases.
  • the nucleic acid is closed-linear DNA (clDNA), i.e. molecules wherein the double stranded region is flanked and protected by two single stranded loops thereby generating dumbbell-shaped molecules.
  • the clDNA consists of a stem region comprising a double stranded DNA sequence of interest covalently closed at both ends by hairpin loops, the clDNA comprising at least two modified nucleotides.
  • clDNA refers to a single stranded covalently closed DNA molecule that forms a “dumbbell” or “doggy-bone” shaped structure under conditions allowing nucleotide hybridization. Therefore, although the clDNA is formed by a single stranded DNA molecule, the formation of the "dumbbell” structure by the hybridization of two complementary sequences within the same molecule generates a structure consisting on a double-stranded middle segment flanked by two single-stranded loops. The skilled in the art know how to generate clDNA from open or closed double stranded DNA using routine molecular biology techniques.
  • a clDNA can be generated by attaching hairpin DNA adaptors, for instance, by the action of a ligase, to both ends of an open double stranded DNA.
  • Hairpin DNA adaptor refers to a single stranded DNA that forms a stem-loop structure by the hybridization of two complementary sequences, wherein the stem region formed is closed at one end by a single stranded loop and is open at the other end.
  • a “modified nucleotide” is any nucleotide (e.g., adenosine, guanosine, cytidine, uracil, and thymidine) that has been chemically modified -by modification of the base, the sugar or the phosphate group- or that incorporates a non-natural moiety in its structure.
  • the modified nucleotide may be naturally or non-naturally occurring depending on the modification.
  • peptide refers to molecules that comprise two or more consecutive amino acids linked to one another via peptide bonds.
  • peptide includes oligopeptides and polypeptides.
  • protein refers to large peptides, in particular peptides having at least about 50 amino acids.
  • peptide and protein are used interchangeably.
  • proteins of interest include, without limitation cytokines, interleukins, tumor necrosis factor (TNF), interferons, integrins, chimeric antigen receptors (CARs), antibodies, hormones, growth factors, enzymes), collagen, fibrinogen, elastin, tubulin, thrombin, serum albumin, erythropoietin, granulocyte colony stimulating factor (G-CSF), colony stimulating factor (CSF), and the like.
  • labeling or imaging agent refers to any substance that is used as a label, or that enhances specific structures in any imaging technique.
  • An imaging agent hence, includes optical imaging agent, magnetic resonance imaging agent, radioisotope, and contrast agent. Imaging or labelling agents are well known in the art.
  • imaging or labelling agents are gases such as sterilized air, oxygen, argon, nitrogen, fluor, perfluorocarbons, carbon dioxide, nitrogen dioxide, sulfur hexafluoride, xenon and helium; commercially available agents used in positron emission tomography (PET), computer assisted tomography (CAT), single photon emission computerized tomography, x-ray, fluoroscopy, and magnetic resonance imaging (MRI).
  • gases such as sterilized air, oxygen, argon, nitrogen, fluor, perfluorocarbons, carbon dioxide, nitrogen dioxide, sulfur hexafluoride, xenon and helium
  • PET positron emission tomography
  • CAT computer assisted tomography
  • single photon emission computerized tomography single photon emission computerized tomography
  • x-ray x-ray
  • fluoroscopy and magnetic resonance imaging
  • MRI magnetic resonance imaging
  • gadolinium chelates currently available, such as diethylene triamine pentaacetic acid (DTPA) and
  • Examples of materials useful for CAT and x-rays include iodine based materials for intravenous administration, such as ionic monomers typified by diatrizoate and iothalamate, non-ionic monomers such as iopamidol, isohexol, and ioversol, nonionic dimers, such as iotrol and iodixanol, and ionic dimers, for example, ioxagalte.
  • Other useful materials include barium for oral use and non-soluble salts such as zinc acetate.
  • an imaging agent is a dye.
  • an imaging agent is a fluorescent moiety.
  • a fluorescent moiety is selected from: a fluorescent protein, a fluorescent peptide, a fluorescent dye, a fluorescent material or a combination thereof.
  • fluorescent dyes include, but are not limited to, xanthenes (e.g., rhodamines, rhodols and fluoresceins, and their derivatives); bimanes; coumarins and their derivatives (e.g., umbelliferone and aminomethyl coumarins); aromatic amines (e.g., dansyl; squarate dyes); benzofurans; fluorescent cyanines; indocarbocyanines; carbazoles; dicyanomethylene pyranes; polymethine; oxabenzanthrane; xanthene; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; but
  • fluorescein dyes include, but are not limited to, 5-carboxyfluorescein, fluorescein-5-isothiocy anate, fluorescein-6-isothiocyanate and 6-carboxyfluorescein.
  • rhodamine dyes include, but are not limited to, tetramethylrhodamine- 6-isothiocyanate, 5-carboxytetramethylrhodamine, 5-carboxy rhodol derivatives, tetramethyl and tetraethyl rhodamine, diphenyldimethyl and diphenyldiethyl rhodamine, dinaphthyl rhodamine, rhodamine 101 sulfonyl chloride (sold under the tradename of TEXAS RED(R)).
  • cyanine dyes include, but are not limited to, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy7, IRDYE680, Alexa Fluor 750, IRDye800CW, ICG.
  • fluorescent peptides include GFP (Green Fluorescent Protein) or derivatives of GFP (e.g., EBFP, EBFP2, Azurite, mKalamal, ECFP, Cerulean, CyPet, YFP, Citrine, Venus, YPet). Fluorescent labels are detected by any suitable method.
  • a fluorescent label may be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence, e.g., by microscopy, visual inspection, via photographic film, by the use of electronic detectors such as charge coupled devices (CCDs), photomultipliers, etc.
  • CCDs charge coupled devices
  • photomultipliers etc.
  • the imaging agent is labeled with a positron-emitting isotope (e.g.,18F) for positron emission tomography (PET), gamma-ray isotope (e.g., 99mTc) for single photon emission computed tomography (SPECT), or a paramagnetic molecule or nanoparticle (e.g.,Gd3+ chelate or coated magnetite nanoparticle) for magnetic resonance imaging (MRI).
  • a positron-emitting isotope e.g.,18F
  • PET positron emission tomography
  • gamma-ray isotope e.g., 99mTc
  • SPECT single photon emission computed tomography
  • MRI magnetic resonance imaging
  • the imaging agent is labeled with: a gadolinium chelate, an iron oxide particle, a super paramagnetic iron oxide particle, an ultra small paramagnetic particle, a manganese chelate or gallium containing
  • gadolinium chelates include, but are not limited to diethylene triamine pentaacetic acid (DTPA), 1 ,4,7,10- tetraazacyclododecane-1 ,4,7,10-tetraacetic acid (DOTA), and 1 ,4,7-triazacyclononane-N,N',N"-triacetic acid (NOTA).
  • the imaging agent is a near-infrared fluorophore for near-infra red (near-IR) imaging, a luciferase (firefly, bacterial, or coelenterate) or other luminescent molecule for bioluminescence imaging, or a perfluorocarbon-filled self-assembled particle for ultrasound.
  • the imaging agent is a nuclear probe. In some molecules, the imaging agent is a SPECT or PET radionuclide probe. In some molecules, the radionuclide probe is selected from: a technetium chelate, a copper chelate, a radioactive fluorine, a radioactive iodine, a indiuim chelate. Examples of Tc chelates include, but are not limited to HYNIC, DTPA, and DOTA.
  • the imaging agent contains a radioactive moiety, for example a radioactive isotope such as 211 At, 13 11, 125 l, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P, 64 Cu radioactive isotopes of Lu, and others.
  • the diagnostically active agents may be attached to the lipo-polyamino acid conjugate backbone in any of A, Ai, A2 or A3 and/or may be contained in the self-assembled particles formed from the conjugated.
  • the present invention also relates to a composition
  • a composition comprising the lipo-polyamino acid conjugate, or alternatively, the self-assembled particle as defined herein, together with one or more appropriate excipients or carriers.
  • the composition is a pharmaceutical composition comprising a therapeutically effective amount of: (a) the lipo-polyamino acid conjugate as defined herein, wherein at least one of A, A1, A2 or A3 is a pharmaceutically active agent or alternatively, (b) a self-assembled particle containing the lipo- polyamino acid conjugate (a), or alternatively, (c) a self-assembled particle which comprises the lipo- polyamino acid conjugate as defined herein and one or more active agents selected from the group consisting of pharmaceutically active agents, nucleic acids, peptides, proteins, and mixtures thereof, together with one or more pharmaceutically acceptable excipients or carriers.
  • therapeutically effective amount refers to the amount of a polymer that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed.
  • the specific dose of the polymer of the invention to obtain a therapeutic benefit may vary depending on the particular circumstances of the individual patient including, among others, the size, weight, age and sex of the patient, the nature and stage of the disease, the aggressiveness of the disease, and the route of administration.
  • the term "pharmaceutically acceptable excipients or carriers” refers to components which are appropriate for use in pharmaceutical technology for the preparation of compositions for medical use. Each component should be “acceptable” in the sense of being compatible with the other ingredients of the composition. When used in contact with the tissue or organ of humans and animals should not have excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • lipo-polyamino acid conjugates described in the present disclosure, and the self-assembled particles and pharmaceutical compositions containing them may be used jointly with other, additional drugs, to provide combined therapy.
  • Said additional drugs may be a part of the same pharmaceutical composition or, alternatively, may be provided in the form of a separate composition for simultaneous or non-simultaneous administration.
  • the composition is a diagnostic composition comprising a diagnostically effective amount of: (a') the lipo-polyamino acid conjugate as defined herein, wherein at least one of A, Ai, A2 or A3 is a diagnostically active agent or alternatively, (b') a self-assembled particle containing the lipo-polyamino acid conjugate (a'), or alternatively, (c') a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more diagnostically active agents, together with one or more diagnostically acceptable excipients or carriers.
  • the term "diagnostic composition” refers to a composition suitable for use in diagnostic, particularly in imaging diagnostic technology.
  • the term “diagnostically effective amount” as used herein, refers to the effective amount of a detection polymer that, when administered, is sufficient for the diagnosis of a disease or disorder; particularly as imaging diagnostic use as contrast imaging agent.
  • the dose of the detection polymer administered will of course be determined by the particular circumstances surrounding the case, including the polymer administered, the route of administration, the particular condition being diagnosticated, and the similar considerations.
  • the diagnostic composition of the present invention comprises one or more diagnostically acceptable excipients or carriers.
  • diagnostically acceptable refers to that excipients or carriers suitable for use in the diagnosing technology for preparing compositions with diagnostic use; particularly by imaging diagnostic use.
  • the detection of these diagnostic agents in the body of the patient can be carried out by the well-known techniques used such as in imaging diagnostic with magnetic resonance imaging (MRI) and X-ray.
  • MRI magnetic resonance imaging
  • the composition is a cosmetic composition comprising a cosmetically effective amount of: (a”) the lipo-polyamino acid conjugate as defined herein, wherein at least one of A, A1, A2 or A3 is a cosmetically active agent or alternatively, (b”) a self-assembled particle containing the lipo-polyamino acid conjugate (a”), or alternatively, (c”) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more cosmetically active agents, together with one or more cosmetically acceptable excipients or carriers.
  • cosmetically effective amount refers to the effective amount of a cosmetically active agent that, when administered, is intended to improve its appearance or to beautify, preserve, condition, cleanse, color or protect the skin, nails or hair without a medical application.
  • cosmetically acceptable or “dermatologically acceptable” excipients or carriers is used interchangeably in this document and refer to components which are appropriate for use in human skin contact without toxicity, incompatibility, instability, inappropriate allergic response, among others.
  • compositions of the invention may be in solid or liquid form.
  • solid forms include frozen forms, lyophilized forms and spray-dried forms.
  • excipients or carriers include, without limitation, diluents, binders, glidants, disintegrants, lubricants colorants, mixtures thereof, and other components known in the state of the art.
  • Any administration route may be used such as e.g. oral, topical, rectal or parenteral route (including subcutaneous, intraperitoneal, intradermal, intramuscular, intravenous route, etc.).
  • the lipo-polyamino acid conjugates of the invention, self-assembled particles, and compositions thereof may be used in therapeutic applications.
  • they may be used as non-viral vectors of general use for biomedical applications, such as vaccines or gene therapy, being effective for transfection of hosts eukaryotic cells in culture, in vivo or ex vivo, monocellular parasites and bacteria, including gene editing using the CRISP/Cas9 methodology.
  • they may be used in protein-based therapy; particularly protein-based vaccine against viral infections or as a therapeutic protein-based vaccine against cancers or infectious diseases.
  • the invention relates to a therapeutic product which is or which comprises: a) a lipo-polyamino acid conjugate of formula (I) as defined herein, wherein at least one of A, Ai, A2 or A3 is a pharmaceutically active agent; or alternatively, b) a self-assembled particle containing the lipo-polyamino acid conjugate a); or alternatively, c) a composition containing the lipo-polyamino acid conjugate a) or the self-assembled particle b); or alternatively, d) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more active agents selected from the group consisting of pharmaceutically active agents, nucleic acids, peptides, proteins, and mixtures thereof; or alternatively, e) a composition containing the selfassembled particle d), for use in medicine.
  • This aspect may also be formulated as a method for treating or preventing a disease or disorder in a subject, more particularly a mammal, and even more particularly a human, comprising administering the therapeutic product as defined herein, and one or more pharmaceutically acceptable excipients or carriers.
  • Non-limiting examples of diseases that may be treated and/or prevented by the derivatives of the present invention include neurodegenerative disorders, neurological diseases, cancer, infectious diseases, disorders related to aging, neuro-inflammation, demyelinating disorders, multiple sclerosis, ischemic disorders, ischemia-reperfusion damage, amyloidotic disease, cardiomyopathy, spinal cord injury, immune disorders, inflammatory disorders, rare diseases, wound healing, skin related diseases and lysosomal storage diseases.
  • Non-limiting examples of neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, cerebral ischaemia, post-encephalitic Parkinsonisms, dystonias, Tourette syndrome, periodic limb movement pathologies, restless legs syndrome, attention deficit hyperactivity disorders, Huntington's disease, progressive supranuclear palsy, Pick's disease, fronto-temporal dementia and neuromuscular diseases.
  • the invention relates to a therapeutic product which is or which comprises: a) a lipo-polyamino acid conjugate of formula (I) wherein any of A, Ai, A2 or A2 is an anticancer agent or an anti-infective agent as defined herein; or alternatively, b) a self-assembled particle containing the lipo-polyamino acid conjugate a); or alternatively, c) a composition containing the lipo-polyamino acid conjugate a) or the self-assembled particle b); or alternatively, d) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more anticancer agents or anti-infective agents; or alternatively, e) a composition containing the selfassembled particle d); for use in the treatment and/or prevention of cancer or an infectious disease, respectively.
  • disorder as used herein is intended to be generally synonymous, and is used interchangeably with, the terms “disease,” “syndrome,” and “condition” (as in medical condition), in that all reflect an abnormal condition of the human or animal body or of one of its parts that impairs normal functioning, is typically manifested by distinguishing signs and symptoms.
  • treat refers to ameliorating symptoms associated with a disease or disorder, including preventing or delaying the onset of the disease or disorder symptoms, and/or lessening the severity or frequency of symptoms of the disease or disorder.
  • the invention relates to the therapeutic product as defined herein for use (i) as transfection reagent for transfecting at least one active agent into a cell; (ii) for use in the in vivo or ex vivo production of biologies encoding a recombinant protein, a peptide or an antibody, or in the production of recombinant virus; (iii) for use as a therapeutic or prophylactic vaccine against viral infections or as a therapeutic vaccine against cancers or infectious diseases; or (iv) for use in genome engineering, for cell reprogramming, for differentiating cells or for gene-editing.
  • the present invention relates to the therapeutic product as defined herein as transfection reagent for delivering one or more nucleic acids (regardless of size and structure, circular and linear nucleic acids) to target cells, in in vivo, in vitro or ex vivo.
  • the active agent is selected from the group consisting of low molecular weight drugs, peptides, proteins, antibodies, nucleic acids, aptamers, and combinations thereof.
  • the present invention also relates to a method for in vitro, ex vivo and in vivo transferring active agents comprising the therapeutic product as defined herein.
  • the therapeutic product is a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more nucleic acids, or alternatively, a composition containing the self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more nucleic acids.
  • transfection reagents of the invention are also useful for co-transfection of two or more active agents simultaneously, e.g. two or more nucleic acids, simultaneously.
  • Transfection compositions (such as kits), as well as methods of using the transfection reagents to deliver nucleic acid to target cells are also within the scope of the present invention.
  • the present invention also provides therapeutic products as defined herein for inducing a regulating effect on the expression of one or more target proteins responsible or involved in genetic hereditary diseases or complex genetic diseases, immune diseases, cancers, viral infections in various tissues/organs or tumors.
  • the present invention also relates to the in vitro or ex vivo use of therapeutic products as defined herein in the production of biologies, in particular biologies encoding a recombinant protein, a peptide or an antibody; or in the production of recombinant virus, such as adeno-associated virus (AAV), lentivirus (LV), adenovirus, oncolytic virus, or baculovirus, or viral or virus-like particles, in particular said products comprising the lipo- polyamino acid conjugates of the invention and at least one nucleic acid molecule for transfection.
  • AAV adeno-associated virus
  • LV lentivirus
  • adenovirus adenovirus
  • oncolytic virus or baculovirus
  • viral or virus-like particles in particular said products comprising the lipo- polyamino acid conjugates of the invention and at least one nucleic acid molecule for transfection.
  • the term "biologies” refers to proteins or nucleic
  • the present invention also relates to an in vitro or ex vivo use of the therapeutic products as defined herein, in particular said products comprising the lipo-polyamino acid conjugates of the invention and at least one nucleic acid molecule for transfection, for genome engineering, for cell reprogramming, for differentiating cells or for gene-editing.
  • compositions for transfecting cells comprise the therapeutic products as defined herein, in particular said products comprising the lipo-polyamino acid conjugates of the invention and at least one nucleic acid molecule for transfection, and an acceptable excipient, buffering agent, cell culture medium, or transfection medium.
  • the present invention is also directed to the therapeutic products as defined herein for use as a therapeutic or prophylactic vaccine against viral infections, or a therapeutic vaccine against cancers.
  • the vaccine is delivered through direct administration such as systemic, intramuscular, intradermal, intraperitoneal, intratumoral, oral, topical, or sub-cutaneous administration, and, in said vaccine, the composition is in association with a pharmaceutically acceptable vehicle.
  • the vaccine can be injected directly into the body, in particular in a human individual, for inducing a cellular and/or a humoral response.
  • the cell targeting is achieved through different mechanisms and depends on the nature and properties of the transfection reagent, method or protocol composition or formulation and the route of administration.
  • the lipo-polyamino acid conjugates of the invention may be used also in cosmetic and diagnostic applications.
  • a diagnostic product which is or which comprises: a') a lipo-polyamino acid conjugate as defined herein, wherein at least one of A, Ai, A2 or A3 is a diagnostically active agent; or alternatively, b') a self-assembled particle containing the lipo-polyamino acid conjugate i'); or alternatively, c') a composition containing the lipo-polyamino acid conjugate i') or the self-assembled particle ii'); or alternatively, d') a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more diagnostically active agents; or alternatively, e') a composition containing the self-assembled particle iv'), for use in diagnostics.
  • This aspect of the invention may also be formulated as the use of a') a lipo-polyamino acid conjugate as defined herein, wherein at least one of A, A1, A2 or A3 is a diagnostically active agent; or alternatively, b') a self-assembled particle containing the lipo-polyamino acid conjugate a'); or alternatively, c') a composition containing the lipo-polyamino acid conjugate a') or the self-assembled particle b'); or alternatively, d') a selfassembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more diagnostically active agents; or alternatively, e') a composition containing the self-assembled particle d'), in diagnostics.
  • a method for the diagnostic of a disease or condition comprising administering a') a lipo-polyamino acid conjugate as defined herein, wherein at least one of A, A1, A2 or A3 is a diagnostically active agent; or alternatively, b') a self-assembled particle containing the lipo- polyamino acid conjugate a'); or alternatively, c') a composition containing the lipo-polyamino acid conjugate a') or the self-assembled particle b'); or alternatively, d') a self-assembled particle which comprises the lipo- polyamino acid conjugate as defined herein and one or more diagnostically active agents; or alternatively, e') a composition containing the self-assembled particle d'), in a subject in need thereof, more particularly a mammal, and even more particularly a human.
  • the invention relates to microbubbles which comprise the lipo-polyamino acid conjugate as defined herein and one or more diagnostically active agents, in particular contrast agents, for use in diagnostics.
  • imaging diagnostic techniques suitable for the present disclosure include, but not limited to, are magnetic resonance imaging (MRI), X-ray, positron emission tomography (PET), singlephoton emission computed tomography (SPECT), fluorescence microscopy, and in vivo fluorescence.
  • MRI magnetic resonance imaging
  • PET positron emission tomography
  • SPECT singlephoton emission computed tomography
  • fluorescence microscopy and in vivo fluorescence.
  • the invention relates to the use in cosmetics of a cosmetic product which is or which comprises: a") a lipo-polyamino acid conjugate as defined herein, wherein at least one of A, Ai, A2 or A3 is a cosmetically active agent; or alternatively, b”) a self-assembled particle containing the lipo-polyamino acid conjugate a'); or alternatively, c”) a composition containing a") or the self-assembled particle b”); or alternatively, d”) a self-assembled particle which comprises the lipo-polyamino acid conjugate as defined herein and one or more cosmetically active agents; or alternatively, e”) a composition containing the selfassembled particle d”).
  • NMR spectra were recorded at 27 °C (300 K) on a 300 UltrashieldTM from Bruker (Billerica MA, USA). Data were processed with the software Topspin (Bruker GmbH, Düsseldorf, Germany). Samples were prepared at a concentration of 20 - 10 mg/mL approx, in the required solvent.
  • Dynamic Light Scattering (DLS) measurements were performed using a Malvern Zetasizer NanoZS instrument, equipped with a 532 nm laser at a fixed scattering angle of 173 °.
  • Polymer solutions were prepared under different conditions (MilliQ water or PBS at different concentrations and temperatures), solutions were sonicated for 10 min and allowed to age for the required time, filtered through a 1.20 pm cellulose membrane filter and measured. Size distribution was measured (diameter, nm) for each polymer per triplicate with n > 3 measurements, automatic optimization of beam focusing and attenuation was applied for each sample.
  • Methionine NCA (2 g, 11.4mmol) was added to a Schlenk tube fitted with a stirring bar and a stopper. After 3 cycles of vacuum/N2, the mixture was dissolved in anhydrous THF (100mg reagents per mL THF). Then, the initiator (i.e. Tetradecylamine 96%, TCI EUROPE N.V., 81 mg, 0.381 mmol) was diluted in THF (5 mL) and added to the reaction mixture, which was stirred at RT for 16 hours. Once NCA consumption was confirmed by IR. Then anhydride acetic (0.359 mL, 3.81 mmol) was added to the reaction mixture and the reaction mixture was stirred at 10°C for 2 hours.
  • THF 100mg reagents per mL THF
  • the initiator i.e. Tetradecylamine 96%, TCI EUROPE N.V.
  • the reaction mixture was poured into diethyl ether to precipitate the product.
  • the precipitate was isolated by centrifugation (3750 rpm, 4 min) and dried under vacuum. Homopolymer was isolated as a white solid. Yield: 70-90%.
  • the reagent will dissolve in DMA (N-butylamine and N-tetradecylamine) or solvent mixtures will be necessary due to the solubility of the initiator: for lipidic initiators such as 1 ,2-dimyristoyl-sn-glycerol-3-phosphoethanol amine, N-ditetradecylamine or ditetradecyl-N- glycine-amine it was necessary to dissolve the initiator in chloroform at a ratio of 1 :3 with respect to the DMA concentration. The mixture was placed into flame-dried Schlenk tube and heated at 60°C for 24 h. The reaction mixture was cooled to room temperature and poured into diethyl ether.
  • DMA N-butylamine and N-tetradecylamine
  • Tocopherol succinate (4.22 mmol) was added to a round bottom flask with a stirrer bar, a stopper and purged with 3 cycles of vacuum/N2, dissolved in DMSCkFW (mL). Then, DMTMM'BF4 coupling agent was added to a round bottom flask with a stirrer bar, a stopper and purged with 3 cycles of vacuum/N2, and dissolved in anhydrous DMSCkFW (mL). DMTMM'BF4 solution was added over tocopherol succinate solution under inert atmosphere and let to react at r.t. for 30 min with stirring.
  • the amphiphile polymer was poured into diethyl ether (1 :10 DMF:diethyl ether) to precipitate the product.
  • the precipitate was isolated by centrifugation (3750 rpm, 4 min) washed two times with diethyl ether and dried under vacuum.
  • the obtained solid was redissolved in methanol at 500 mg/mL and then the solution was poured again into diethyl ether (1 :10 to methanokdiethyl ether), washed two times with diethyl ether and dried under vacuum at least for 2 hours.
  • the solid was dissolved in distilled water and diafiltrated in a tangential flow filtration system with the addition of some drops of HCI for total DIPEA purification.
  • oligonucleotides used were: a mRNA purchased from Trilink, with reference L- 1201-1000 CleanCap Flue mRNA (5moU) expressing luciferase as reporter gene. Any other oligonucleotide with the characteristics above indicated could be used to carry out the experiments below.
  • the ionizable lipid (6Z,9Z,28Z,31Z)-heptatriacont-6,9,28,31-tetraene-19-yl 4-(dimethylamino)butanoate (DLin- MC3-DMA) was purchased from Nanosoft Polymers, or [(4-Hydroxybutyl)azanediyl]di(hexane-6, 1-diyl) bis(2- hexyldecanoate) (ALC-0315) was purchased from BroadPharmShielding lipid 1 ,2-Dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG2000) was purchased from Avanti Polar Lipids.
  • LNPs Both polymers were used to form benchmark LNPs.
  • Structural lipids used to form LNPs were: 1 ,2-distearoyl-sn-glycero-3- phosphocholine (DSPC, from Avanti Polar Lipids, 850365P) and Cholesterol (Sigma-Aldrich, C8667).
  • a microfluidic HerringboneMixer from Darwing Microfluidics was used for the formulations of LNPs 1-2.
  • the reactor presents 2 inlet-channels (one for the DNA in aqueous phase and the other for the lipid mixture in ethanol) and 1 outlet-channel.
  • the specifications of the mixer are the following:
  • two programmable pumps control the fluid flow rates of the syringes (NE-1000 Programmable Single Syringe Pump, Syringe Pump, USA).
  • the system accepts infusion rates from 0.73 piL/h (1 mL syringe) to 2100 mL/h (60 mL syringe). This methodology provides reproducibility to the formation of LNPs as well as the possibility of scaling up the process.
  • LNPs were diluted in 10 mM Phosphate buffer saline (PBS) pH 7.4 and concentrated with centrifugal concentrators (VivaspinTM 500, Sigma) to remove ethanol
  • the formulations containing the lipo-polyamino acid conjugates of formula (I) were prepared analogously with the difference that lipo-polyamino acid conjugates of formula (I) were used instead of DMG-PEG and the lipid ratio was different (shown below in Example 7).
  • LNPs 3 to 7 were formulated with NanoScaler (KNAUER) as follows:
  • Lipids were dissolved in ethanol at 46.3/9.4/42.7/1.6 molar ratio (ALC-0315/DSPC/Cholesterol/ DMG- PEG).
  • LNPs 3 to 7 were further diluted in 10 mM Phosphate buffer saline (PBS) pH 7.4 to decrease ethanol concentration and purified with centrifugal concentrators (VivaspinTM 500, Sigma) to remove ethanol.
  • PBS Phosphate buffer saline
  • the formulation containing the polymer of the invention was prepared analogously with the difference that different polymers was used instead of shielding lipid DMG-PEG and the lipid ratio was different (shown below in Example 7).
  • DLS is equipped with a 660 nm laser diode and 2 piL of the samples were added in a Stunner plate with a microfluidic circuit, which has two fixed pathlengths that cover a wide dynamic range of 0.03-275 OD. Size distribution was measured (diameter, nm) with n > 3 measurements.
  • Table 9 shows the results obtained of PMetOI formulated at molar ratio of 5% containing pDNA as cargo.
  • Results show when using 5% of polymer the results were consistent in all replicates, showing low PDI values and appropriate sizes.
  • This composition was also used to formulate LNPs with different mRNA (Firefly luciferase (luc); RBD and GFP) obtaining low PDI values and appropriate sizes (Table 10-13).
  • Results show when using 2 and 5% of polymer at different compositions the results were consistent in all replicates, showing low PDI values and appropriate sizes.
  • LNPs apparent pKa values were determined by measuring the fluorescence of 2-(p-toluidino)-6-napthalene sulfonic acid (TNS) during titration with different buffer solutions: from pH 4 to 5.5 (0.5 pH unit increments) with 20 mM Acetate buffer; from pH 6 to 7.5 (0.5 pH unit increments) with 20 mM Phosphate buffer; from pH 8 to 9 (0.5 pH unit increments) with 20 mM Borate buffer.
  • TMS 2-(p-toluidino)-6-napthalene sulfonic acid
  • the formulation was diluted to 20 piM lipid concentration with different buffer solutions and added to a black 96-well plate.
  • TNS solution 0.3 mM in DMSO
  • Example 8 In vitro biological studies in HEK293 cells.
  • HEK293 cells were cultured in DMEM high glucose with Glutamax (Gibco- Thermo Fisher # 61965-059) supplemented with 10% of Fetal Bovine Serum (Hyclone # SV30160.03HI, provided by GE Healthcare Europe GmbH) and 1 % penicillin/streptomycin. Transfections were carried out on 96-well plates containing 10OOOcells/well in a final volume of 10Opil, and cells were incubated 24 hours at 37°C and 5% CO2. After 24h from cell seeding, the medium was removed and refreshed with 90pil of complete medium. 10pil of each LNPs formulation were added to the cells. In the case of positive control (LipofectamineTM 2000 Transfection Reagent, I nvitrogen TM), manufacturer guidelines were followed. After 24 hours cells were recovered and processed.
  • ATPLite reagent ATPLite PerkinElmer #6016731
  • Luminiscence was read spectrophotometrically using VictorNivo (PerkinElmer) following manufacturer's instructions and data was represented as the percentage of cell viability, taken untreated control cells as 100%.
  • the transfection efficiency and the cell viability of the LNPs formulations in HEK293 cells is reported in the following table.
  • the transfection data is represented as % of the positive control DMG-PEG with the formulation reported 46.3/9.4/44.3-x/x molar ratios for ALC-0315/DSPC/Cholesterol/DMG-PEG or PMetOn being x the different molar ratios for each PMetOn employed, and being the positive control 100% after 24h of treatment and cell viability is compared to non-treated (NT) cells, being the ATP content readout of NT (nontreated) cells equal to 100%.
  • NT non-treated
  • mice were injected subcutaneously Luciferin (15 mg/kg), and after 10 minutes mice were euthanized via CO2 inhalation and organs dissected (blood, liver, spleen, lungs, heart, kidney and ALNs (Axillary lymph nodes)).
  • Image acquisition was obtained by MS Spectrum (settings: Binning factor 8 F/Stop 1 Exp AUTO FOV 22,5 (D)). The weight of the different organs was obtained and then freezed with N2 and -80°C.
  • Image Analysis was performed by Living Image 4.5.4 (settings: ROI (Draw auto, threshold 2%) and data represented in total counts I tissue g.
  • mice after administration of the different formulations were weighted obtaining the following results:
  • mice Body and organs weight (g) of mice administered iv with the formulations at time 4 and 24h.
  • mice body weights remained in the range of 20-23 g, showing no significant differences. Same behavior was observed for all the organs, showing no significant differences between non-treated (PBS) and LNP 12 with mice administered with LNP 4.
  • Clause 4 The lipo-polyamino acid conjugate according to any of clauses 1 to 3, wherein m is an integer from 5 to 150, more particularly from 5 to 100, even more particularly from 5 to 50, and even more particularly from 5 to 30 or from 5 to 25, particularly wherein m is measured by 1 H-NMR spectroscopy.
  • Clause 7 The lipo-polyamino acid conjugate according to any of clauses 1 to 5, wherein in the PAA2 repetitive units of formula (III), R5 is -(Ci-Cejalkyl, more particularly R5 is -CH3.
  • n is an integer from 5 to 150, more particularly from 5 to 100, even more particularly from 5 to 50, and even more particularly from 5 to 30 or from 5 to 25, particularly wherein m is measured by 1 H-NMR spectroscopy.
  • Clause 14 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is N and Ri is H.
  • Clause 15 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is N and Ri is -(Ci-Cisjalkyl, more particularly -(Ci-Ci2)alkyl.
  • Clause 16 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is N and Ri is -(C2-Ci8)alkenyl, more particularly -(C2-Ci2)alkenyl.
  • Clause 17 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is O and Ri is absent.
  • Clause 18 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is S and Ri is absent.
  • Clause 19 The lipo-polyamino acid conjugate according to any of clauses 1 to 13, wherein X is CH and Ri is H, -(Ci-Cisjalkyl, or -(C2-Ci8)alkenyl, more particularly X is CH and Ri is H, -(Ci-Ci2)alkyl, or -(C2-Ci2)alkenyl, and even more particularly X is CH and Ri is H.
  • Clause 20 The lipo-polyamino acid conjugate according to any of clauses 1 to 19, wherein R2 is a radical of formula (IV), more particularly, the radical of formula (IV) is selected from the formulas (iv1)-(iv24) as defined herein.
  • Clause 21 The lipo-polyamino acid conjugate according to any of clauses 1 to 19, wherein R2 is a radical of formula (V), more particularly, the radical of formula (V) is selected from the formulas (v1)-(v24) as defined herein.
  • Clause 25 The lipo-polyamino acid conjugate according to any of clauses 1 to 23, wherein R3 is -(Ci-Cejalkyl, more particularly R3 is -CH3.
  • Clause 26 The lipo-polyamino acid conjugate according to any of clauses 1 to 25, wherein A is a lipid-like moiety R7, and A1 is other than a lipid-like moiety Rz'.
  • Clause 28 The lipo-polyamino acid conjugate according to any of clauses 1 to 26, wherein the lipid-like moiety Rz is -(C2-C2o)alkenyl, more particularly -(Ce-C jalkenyl, and even more particularly, either - (Ci2-Cis)alkenyl or a radical of the formula (XXIV) as defined herein.
  • Clause 32 The lipo-polyamino acid conjugate according to any of clauses 1 to 26, wherein the lipid-like moiety Rz is a radical of formula (X) as defined herein.
  • Clause 33 The lipo-polyamino acid conjugate according to any of clauses 1 to 26, wherein the lipid-like moiety Rz is a radical of formula (XI), more particularly, a radical selected from the formulas (xi1), (xi2), (xi3), (xi4), (xi5), (xi6), (xi7) and (xi8), as defined herein.
  • Clause 42 The lipo-polyamino acid conjugate according to any of clauses 1 to 10 or 12 to 41, wherein p is 0, and Ai is H.
  • Clause 43 The lipo-polyamino acid conjugate according to any of clauses 1 to 10 or 12 to 41, wherein p is 0, and Ai is an amino protective group.
  • Clause 45 The lipo-polyamino acid conjugate according to any of clauses 1 to 10 or 12 to 41, wherein p is 0, and Ai is selected from the group consisting of H, -(Ci-Cejalkyl, -(Cs-C jaryl, -(C5-Cio)heteroaryl, -(Ce- Cio)aralkyl, -(Ci-C6)alkyl-0-(C5-Cio)aryl, and - (C5-Cio)heterocycloalkyl; being Ai optionally substituted as defined herein.
  • Clause 46 The lipo-polyamino acid conjugate according to any of clauses 1 to 9 or 11 to 41, wherein p is 1, and Ai is selected from the group consisting of H, -OH, -(Ci-Cejalkyl, -(Cs-C jaryl, -(C5-Cio)heteroaryl, -(C6-Cio)aralkyl, -(Ci-C6)alkyl-C-(C5-Cio)aryl, -(C5-Cio)heterocycloalkyl, -(C1-C10) alkoxy, -(C6-Cio)aryloxy, -(C6-Cio)aralkoxy, -(Cs-Ciojheteroaralkoxy, -(Ci-Cio)alkyl-C-(C6-Cio)aryloxy; being Ai optionally substituted as defined herein.
  • Clause 48 The lipo-polyamino acid conjugate according to any of clauses 1 to 9 or 11 to 41, wherein p is 1 , and Ai is a lipid-like moiety R/.
  • Clause 50 The lipo-polyamino acid conjugate according to any of clauses 1 to 25, wherein Ai is a lipid-like moiety R/ and A is other than a lipid-like moiety R7.
  • Clause 53 The lipo-polyamino acid conjugate according to any of clauses 1 to 25, 47 to 48 or 50, wherein the lipid-like moiety R7 is -(C2-C2o)alkenyl, more particularly -(Ce-Cisjalkenyl, and even more particularly, a radical of the formula (XXIV) as defined herein.
  • Clause 54 The lipo-polyamino acid conjugate according to any of clauses 1 to 25, 47 to 48 or 50, wherein the lipid-like moiety Rz’ is a radical of formula (XXIV) as defined herein.
  • Clause 66 The lipo-polyamino acid conjugate according to any of clauses 1 to 25, 47 to 48 or 50, wherein the lipid-like moiety Rz is selected from the group consisting of -(Ci-Ci8)alkyl, -(C2-Ci8)alkenyl, and a radical of formula (XI), (XII), (XIV), (XV), (XVI), (XVII) or (XVIII’).
  • Clause 67 The lipo-polyamino acid conjugate according to any of clauses 1 to 9, 11 to 25 or 42 to 66, wherein p is 1, and A is H.
  • Clause 68 The lipo-polyamino acid conjugate according to any of clauses 1 to 9, 11 to 25 or 42 to 66, wherein p is 1, and A is a protective group.
  • Clause 69 The lipo-polyamino acid conjugate according to any of clauses 1 to 9, 11 to 12, 14 to 25 or 42 to 66, wherein p is 1, s is 0 and A is selected from the group consisting of H, -(Ci-Cejalkyl, -CO(Ci- Cejalkyl, -(Ci-C6)alkyl-CO-N[(Ci-Ci8)alkyl]2, -(Cs-C jaryl, -(Cs-Ciojheteroaryl,
  • Clause 70 The lipo-polyamino acid conjugate according to any of clauses 1 to 9, 11 to 25 or 42 to 66, wherein p is 1, and A is selected from the group consisting of a pharmaceutically active agent, a cell-targeting agent, a penetration enhancing agent, a cosmetically active agent, and a diagnostically active agent.
  • X is N or CH
  • A is selected from the group consisting of butyl, (C12-C18) alkyl, a radical of formula (XXV), and a radical of formula (XXVI);
  • Ri is selected from the group consisting of H and (Ci-Cis)alkyl;
  • R” is selected from the group consisting of H, -CO(Ci-C6)alkyl, a radical of formula (XVIII”), and -CONH(Ci-Ci8)alkyl;
  • R2 is as defined herein, particularly R2 is a radical of formula (VI) or a radical of formula (VI), wherein more particularly in the radical of formula (VI), c is an integer from 0 to 3;
  • R is -(Ci- Cejalkyl, and R' is -(Ci-C6)alkyl-CC>2'.
  • R4 is selected from the group consisting of H, methyl; isopropyl, isobutyl, and sec-butyl; n is a value from 5 to 200, or from 5 to 150, or from 10 to 150, particularly measured by 1 H- NMR spectroscopy; and m is 0 or a value from 5 to 150, or from 5 to 100, or from 5 to 80, or from 10 to 80, particularly measured by 1 H-NMR spectroscopy.
  • a self-assembled particle comprising the lipo-polyamino acid conjugate of formula (I) as defined in any of clauses 1-71, and optionally one or more active agents selected from the group consisting of pharmaceutically active agents, cell-targeting agents, penetration enhancing agents, cosmetically active agents, diagnostically active agents, nucleic acids, peptides, proteins, and mixtures thereof.
  • Clause 73 The self-assembled particle according to clause 72, and one or more nucleic acids.
  • Clause 74 The self-assembled particle according to any of the clauses 72 to 73, wherein the self-assembled particle is a nanoparticle, more particularly selected from the group consisting of a micelle, an inverted micelle, a planar bilayer, a crystal nanoparticle, a liposome, microbubbles, and a lipid nanoparticle, and even more particularly, the self-assembled particle is a lipid nanoparticle or a liposome.
  • the self-assembled particle according to any of the clauses 72 to 74 which further comprises one or more lipids selected from the group consisting of ionisable lipids, cationic lipids, neutral lipids, and anionic lipids.
  • Clause 76 The self-assembled particle according to any of the clauses 72 to 74, which further comprises a ionizable lipid or a cationic lipid, a phospholipid, and a sterol.
  • the self-assembled particle according to clause 76 which is particularly a lipid nanoparticle or a liposome and which comprises: i) the lipo-polyamino acid conjugate of formula (I) as defined in any of clauses 1-70, or any stereoisomer or mixtures of stereoisomers, either of the compound of formula (I), or of any of its salts, in an amount from 0.1 to 10 mol%; ii) a ionizable or a cationic lipid in an amount from 30 to 70 mol%; iii) a phospholipid in an amount from 1 to 20 mol%; iv) a sterol in an amount from 20 to 60 mol%; and wherein the percentages are expressed with respect to the sum of the mol% of the lipids and the lipopolyamino acid conjugate of formula (I);

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Abstract

L'invention concerne des conjugués de lipo-polyaminoacides contenant du soufre de formule (I), ainsi que des sels, stéréoisomères et mélanges acceptables de ceux-ci. L'invention concerne également des particules auto-assemblées comprenant ces conjugués lipo-polyaminoacides et éventuellement des agents actifs, et des compositions comprenant les conjugués lipo-polyaminoacides ou les particules auto-assemblées les comprenant. L'invention concerne également l'utilisation des conjugués de lipo-polyaminoacides, des particules auto-assemblées, ou des compositions les comprenant pour la médecine, la cosmétique et le diagnostic, et l'utilisation des conjugués de lipo-polyaminoacides de formule (I) en tant que vecteurs.
PCT/EP2024/072025 2023-08-04 2024-08-02 Lipo-polyaminoacides contenant du soufre stabilisés et furitfs Pending WO2025031991A1 (fr)

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