Poly (oxazoline) and poly (oxazine) based lipids, methods for the preparation thereof and uses thereof

Abstract

The present invention relates to poly (oxazoline) and poly (oxazine)-based lipids, to a process for the preparation thereof and to the use thereof. Disclosed are polymers of the formula Ini-[N (COR1)-CR3H-CR4H] w-R2 (I), Ini-[N (COR1)-CR3H-CR4H] w-R2 (II), as well as biodegradable derivatives thereof obtained by hydrolysis and oxidation. The group R2 is selected from the group consisting of:-OR11,-OCO-R11,-OCO-R14-CO-OR11,-OCO-R14-CO-NR12R13,-NR12-R14-CO-NR13R15,-O-R16-(O-OC-R18) m, and-(cyclo-N3R17)-CH2-OCO-NR12R13, where R11, R13, and R18 correspond to a C6-C20 alkyl group, and a method for preparing the same. These polymers are prepared by modification of the omega-terminal groups of telechelic poly (2-n-alkyl-oxazolines) and poly (2-n-alkyl-oxazines), have amphiphilic properties and are suitable for use as substitutes for polyethylene glycol, in particular in active ingredient formulations.

Description

Technical Field

[0001] The present invention relates to novel polymeric lipids which are suitable as replacements for polyethylene glycol (hereinafter also referred to as "PEG"). Furthermore, the present invention relates to the preparation of these polymers and their use in the preparation of pharmaceutical formulations. Background Art

[0002] Biocompatible polymers are highly attractive materials for biomedical applications, such as for drug administration. PEG is widely used in pharmaceutical products due to the advantages associated with their use. For example, in SARS-CoV-2 mRNA vaccines, lipid nanoparticles containing PEG lipids as key components are used to deliver mRNA. In these lipid nanoparticles, PEG lipids not only affect the particle size during preparation, but also prevent the aggregation of particles and contribute to their storage stability. In addition, PEG prolongs the circulation time of particles in the blood due to its stealth effect, thus preventing rapid recognition and clearance by the immune system (see X. Hou, T. Zaks, R. Langer, Y. Dong, Nat. Rev. Mater. 2021, 6, 1078-1094).

[0003] However, so-called PEGylation also brings significant disadvantages, which are called the "PEG dilemma". Due to the stimulation of anti-PEG antibodies, which are widely spread in humans due to the excessive use of PEG in cosmetics, accelerated clearance occurs in the blood, so that the PEGylated particles cannot effectively reach their desired site of action, which leads to lower efficacy. In addition to lower transfection efficiency, anti-PEG antibodies may also cause hypersensitivity reactions in humans that manifest as pseudoallergies, for example in SARS CoV 2 mRNA vaccines (see T. Ishida, M. Ichihara, X. Wang, K. Yamamoto, J. Kimura, E. Majima, H. Kiwada, J. Controlled Release 2006, 112, 15-25 and S. S. Nogueira, A. Schlegel, K. Maxeiner, B. Weber, M. Barz, M. A. Schroer, C. E. Blanchet, D. I. S. Vergun, S. Ramishetti, D. Peer, P. Langguth, U. Sahin, H. Haas, ACS Appl. Nano Mater. 2020, 3, 10634-10645).

[0004] In addition to these disadvantages, another problem in the use of PEG is the formation of toxic byproducts such as 1,4-dioxane during the synthesis and the formation of PEG oligomers by subsequent oxidation when using PEG with a lower molar mass (see K. Knop, R. Hoogenboom, D. Fischer, US Schubert, Angew. Chem. Int. Ed. 2010, 49, 6288-6308). It is therefore important to provide PEG alternatives.

[0005] Poly(2-n-alkyl-2-oxazoline) with short side chains (hereinafter also referred to as "PAOx") exhibit similar hydrophilicity, biocompatibility and "stealth effect" and therefore appear to be promising candidates for replacing PEG, which is further confirmed in a detailed comparison of their dissolution behavior (see M. Grube, MN Leiske, US Schubert, I. Nischang, Macromolecules 2018, 51, 1905-1916). Compared with PEG, PAOx also exhibit greater structural diversity due to their side chain modifiability.

[0006] PAOx with longer side chains are hydrophobic and can be used to prepare amphiphilic copolymers, low surface energy materials or low adhesion coatings. Thermal and crystallization properties can also be adaptively adjusted by changes in the PAOx side chains (see R. Hoogenboom, MWM Jijten, HML Thijs, BM van Lankvelt, US Schubert, Designed Monomers and Polymers 2005, 8, 659-671; EFJ Rettler, JMK Ranenburg, HM

[0007] L.Lambermont-Thijs, R.Hoogenboom, USSchubert, Macromolecular Chemistryand Physics 2010, 211, 2443-2448; K.Kempe, M.Lobert, R.Hoogenboom, USSchubert, Journal of Polymer Science Part A: Polymer Chemistry 2009,47,3829-3838; M.Beck, P.Birnbrich, U.Eicken, H.Fischer, WEFristad, B.Hase, HJKrause, Die AngewandteMakromolekulare Chemie 1994,223,217-233;JMRodriguez-Parada, M.Kaku, DYSogah, Macromolecules 1994,27,1571-1577; N.Oleszko-Torbus, A. M.Bochenek, D.Lipowska-Kur, A.Dworak,W. Polymer Chemistry 2020, 11, 15-33; AL Demirel, P. Tatar Güner, B. Verbraeken, H. Schlaad, US Schubert, R. Hoogenboom, Journal of Polymer Science Part B: Polymer Physics 2016, 54, 721-729). Schubert and co-workers previously reported that for a series of poly(2-n-alkyl-2-oxazoline)s, the side chain length continued to increase until poly(2-pentane-2-oxazoline) and the glass transition temperature (T g ) decreases. For PAOx with longer side chains, a melting temperature T that is independent of the side chain length is observed. m crystallization properties.

[0008] Polyoxazolines PAOx (of which poly(2-ethyl-2-oxazoline) is of particular interest) therefore appear to be alternatives to PEG, as they also have the stealth effect of PEG. It is speculated that PAOx lipids could be alternatives to PEG lipids, for example to PEG lipid ALC-0159, which is used in the BioNTech mRNA vaccine

[0009] There have been studies synthesizing PEG lipid substitutes. S.NOGUEIRA et al. cooperated with BioNTech to prepare lipids from polysarcosine (pSar). However, these pSar lipids show lower transfection than PEG-lipid references, which makes them unattractive substitutes for vaccination (see SSNogueira, A.Schlegel, K.Maxeiner, B.Weber, M.Barz, MASchroer, CEBlanchet, DISvergun, S.Ramishetti, D.Peer, P.Langguth, U.Sahin, H.Haas, ACS Appl.Nano Mater.2020,3,10634-10645).

[0010] M. BENTLEY et al. synthesized lipids from polyoxazolines in which phospholipids were coupled to PAOx (see US Pat. No. 9,284,411 B2 and US Pat. No. 8,883,211 B2).

[0011] PAOx and PEG are not biodegradable, but for many biomedical applications it is important to prevent the long-term accumulation of polymers with higher molecular mass.

[0012] Therefore, it is desirable to have a lipid based biodegradable polyoxazoline (hereinafter also referred to as "degPAOx") to ensure biodegradability. The synthesis of degPAOx has been reported (see NE M. Kleinsteuber, C. Weber, US Schubert, Macromolecules 2020, 53, 10837-10846) and WO2022 / 106049 A1.

[0013] Functionalized polyglycine-poly(alkyleneimine) copolymers are known from WO 2022 / 106049 A1. Polymers having long-chain alkyl groups or having long-chain alkyl ester groups as end groups are not disclosed in this document.

[0014] The use of PAOx lipids and degPAOx lipids is not limited to vaccine applications, but these lipids can be generally used as carrier materials for drug or gene delivery.

[0015] Using analytical ultracentrifugation, the hydrodynamic radius of PEG-lipid surrogates can be measured. In this way, the molar mass of PAOx lipids and degPOx lipids can be accurately matched to the hydrodynamic volume of commercially available PEG types, such as commercially available PEG lipid ALC-0159, which facilitates the potential replacement of PEG lipids with PAOx lipids and degPAOx lipids in existing biomedical applications. Summary of the invention

[0016] It is therefore an object of the present invention to provide new polymeric lipids which are suitable as alternatives to PEG lipids.

[0017] Another object of the present invention is to provide a simple method for preparing these polymeric lipids.

[0018] The object is solved by providing a first group of polymers of formula (I) or (II),

[0019]

[0020] Alternatively, a second group of polymers is provided, which, based on all structural units, contains 10 to 95 mol % of structural units of formula (III), 5 to 90 mol % of structural units of formula (IV) and 0 to 20 mol % of structural units of formula (V)

[0021]

[0022] or a compound containing 10 to 95 mol % of a structural unit of formula (VI), 5 to 90 mol % of a structural unit of formula (VII) and 0 to 20 mol % of a structural unit of formula (VIII)

[0023]

[0024] Which has the structural part -CHR 4 -、-CHR 5 - or CHR 7 - or having the structural part -CHR 5 -、-CHR 7 -or-CHR 10 - at least one of the structural units of formula (III), (IV) or (V) or formula (VI), (VII) or (VIII) with the group R 2 Covalent bonding,

[0025] in

[0026] Ini is a group derived from a cationic polymerization initiator,

[0027] R 1 Selected from the group consisting of: hydrogen or C1-C4 alkyl,

[0028] R 2 Selected from the group consisting of: -OR 11 、-OCO-R 11 、-OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 、-NR 12 -R 14 -CO-NR 13 R 15 、-OR 16 -(O-OC-R 18 ) m and

[0029]

[0030] R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 independently of one another represent hydrogen, methyl, ethyl, propyl or butyl,

[0031] R 11 It is C6-C 20 alkyl,

[0032] R 12 and R 15 are independently hydrogen or alkyl,

[0033] R 13 Indicates C6-C 20 alkyl,

[0034] R 14 represents an alkylene group, a cycloalkylene group, an arylene group or an aralkylene group,

[0035] R 16 is an m+1 valent aliphatic hydrocarbon group,

[0036] m is an integer from 1 to 5,

[0037] R 18 It is C6-C 20 Alkyl, provided that the group R 16 Several groups R 18 can be different within a given definition,

[0038] R 17 is a trivalent bicyclic radical, and

[0039] w is an integer in the range of 1-5000.

[0040] In the context of this specification, "polymer" is understood to be an organic compound as mentioned above, characterized by the repetition of specific units (monomer units or repeating units). Polymers can be composed of one type or several types of different repeating units. Polymers are prepared (polymerized) by chemical reactions of monomers to form covalent bonds and form a so-called polymer backbone by connecting the polymerized units. This may have side chains on which functional groups may be located. If a part of the polymer has hydrophobic properties, they can form nanoscale structures (e.g. nanoparticles, micelles, vesicles) in an aqueous environment. Homopolymers consist of only one monomer unit. On the other hand, copolymers consist of at least two different monomer units, which may be randomly distributed, distributed as a gradient, alternately distributed or distributed as blocks.

[0041] The polymer according to the present invention is a functionalized poly(oxazoline) or poly(oxazine). The former is derived from oxazoline and the latter from oxazines. The following description focuses primarily on the preparation and use of poly(oxazoline). These explanations also apply flexibly to homologous poly(oxazines).

[0042] In the context of this specification, "lipid" is understood to mean a substance that is completely or at least mostly insoluble in water (hydrophobic) and well soluble in hydrophobic (lipophilic) solvents. Lipids are amphiphilic and represent a subclass of surfactants. In polar solvents such as water, lipids often form micelles, vesicles or membranes.

[0043] In the context of this specification, "active ingredient" means a compound or mixture of compounds that exerts a desired effect on a living organism. These can be, for example, pharmaceutically active ingredients or agrochemically active ingredients. The active ingredient can be a low or high molecular weight organic compound. Preferably, the active ingredient is a low molecular weight pharmaceutically active substance or a higher molecular weight pharmaceutically active substance, wherein hydrophilic active ingredients from potentially therapeutically useful nucleic acids (e.g., short interfering RNA, short hairpin RNA, microRNA, messenger RNA, plasmid DNA) or from potentially useful proteins (e.g., antibodies, interferons, cytokines) can be used in particular. Preferred examples of active ingredients are vaccines or nucleic acids.

[0044] The active ingredients here may be those which, without inclusion in nanoparticles or liposomes, have only low or no bioavailability, low or no in vivo stability, or are intended to act only in specific cells of the organism.

[0045] In the context of this specification, "excipients and additives" should be understood as meaning to be added in the preparaton to give it specific additional properties and / or facilitate the material of its processing. The example of excipients and additives is tracer, contrast agent, carrier, filler, pigment, dye, spices, lubricant, UV stabilizer, antioxidant or surfactant. Especially, "excipients and additives" should be understood as any pharmacologically compatible and therapeutically useful material, which is not a pharmaceutically active ingredient, but it can be prepared in a pharmaceutical composition together with a pharmaceutically active ingredient, so as to affect, particularly improve the qualitative properties of a pharmaceutical composition. Preferably, excipients and / or additives do not play or do not play an obvious pharmacological effect or at least do not play an undesirable pharmacological effect about the expected treatment.

[0046] Ini is a residue derived from an initiator of cationic polymerization leading to the formation of a poly(oxazoline). Ini may be an organic group such as an alkyl, cycloalkyl, aryl, aralkyl or heterocyclic group. However, other groups are also conceivable. Examples of such groups can be found in US 8,883,211 B2.

[0047] Group R 12 , R 15 Ini and Ini may represent an alkyl group. These are typically alkyl groups having from one to twenty carbon atoms, which may be straight-chain or branched. Examples thereof are methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl or eicosyl.

[0048] Group R 11 , R 13 and R 18 Can represent C6-C 20 Alkyl. These are alkyl groups having six to twenty carbon atoms, which may be straight-chain or branched. Examples are hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl or eicosyl.

[0049] Group R 1 C1-C4 alkyl may be represented. These are alkyl groups having one to four carbon atoms, which may be straight-chain or branched. Examples are methyl, ethyl, propyl and butyl.

[0050] Preferably, R 1 It is methyl, ethyl or propyl, and methyl or ethyl is particularly preferred.

[0051] The group Ini may represent a cycloalkyl group. These are generally cycloalkyl groups having five to six ring carbon atoms. Cyclohexyl is particularly preferred.

[0052] The group Ini may represent an aryl group. These are generally aromatic hydrocarbon groups having five to ten ring carbon atoms. Phenyl is preferred.

[0053] The group Ini may represent an aralkyl group. These are typically aryl groups which are linked to the rest of the molecule via an alkylene group. Benzyl is preferred.

[0054] The group Ini may represent a heterocyclic group. These are generally aromatic or nonaromatic hydrocarbon groups having five to ten ring carbon atoms, which have one or two heteroatoms in the ring, for example nitrogen and / or oxygen and / or sulfur.

[0055] Group R 14 " may represent an alkylene radical. These are generally alkylene radicals having one to six carbon atoms, which may be straight-chain or branched. Examples of alkylene radicals are methylene, ethylene, propylene, butylene, pentylene and hexylene. Ethylene, propylene and butylene are preferred and ethylene is particularly preferred.

[0056] Group R 14 may represent cycloalkylene. These are typically cycloalkylene groups having five to six ring carbon atoms. Cyclohexylene is particularly preferred.

[0057] Group R 14 may represent an arylene radical. These are generally divalent aromatic hydrocarbon radicals having five to ten ring carbon atoms. Phenylene is preferred.

[0058] Group R 14 may represent aralkylene. These are typically arylene groups with an alkylene group, wherein the connection of the aralkylene group to the rest of the molecule occurs via the arylene group and the alkylene group. Benzylidene is preferred.

[0059] R 16 is a divalent to hexavalent (m+1 valent) aliphatic hydrocarbon radical derived from an m+1 valent aliphatic alcohol. One of the OH oxygen atoms of this alcohol is covalently bonded to the polyoxazoline. The remaining OH groups of this alcohol are esterified with fatty acids. If multiple ester groups are present in the radical, these can each be derived from the same or different fatty acids. Examples of divalent alcohols are ethylene glycol or propylene glycol; examples of trivalent alcohols are glycerol or trimethylolpropane; examples of tetravalent alcohols are pentaerythritol; and examples of hexavalent alcohols are sugar alcohols. Preferably, R 16 It is a group derived from glycerol.

[0060] Group R 17It is a trivalent bicyclic group. These are usually trivalent groups consisting of two cycloalkyl groups, one of which has three annular carbon atoms and the other of which has five to eight annular carbon atoms. The larger rings in these rings contain double bonds. Connection to the rest of the molecule is achieved via a covalent bond derived from the group with the three annular carbon atoms and via two other covalent bonds derived from the group with the five to eight annular carbon atoms.

[0061] A first group of polymers according to the invention are linear polymers.

[0062] A second group of polymers according to the invention can be linear or branched polymers. Linear polymers are preferred here.

[0063] The second group of linear polymers has the structure of formula (IX) or (X)

[0064]

[0065] Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 has the meaning defined above,

[0066] x and y are independently integers in the range of 1-5000,

[0067] z is an integer in the range 0-1000, provided that

[0068] Depend on[] x The mole fraction of the structural unit marked is 10 to 95 mol%, represented by [] y The mole fraction of the structural unit marked is 5 to 90 mol%, and is represented by [] z The mole fraction of the structural units indicated is from 0 to 20 mol %, in each case based on the formula [] x , [] y and[] z The total amount of labeled structural units.

[0069] The solubility of the polymers according to the invention can be influenced by copolymerization with suitable monomers and / or by functionalization. Such techniques are known to those skilled in the art.

[0070] The polymers according to the invention may comprise a wide range of molar masses. Typical molecular weights (M n) can vary in the range of 1,000-500,000 g / mol, in particular 1,000-50,000 g / mol. These molar masses can be determined by the dissolved polymer 1 H-NMR spectroscopy. Specifically, the molar mass can be determined using analytical ultracentrifugation or chromatographic methods such as size exclusion chromatography.

[0071] Preferred polymers according to the invention have an average molar mass (number average) in the range of 1,000 to 50,000 g / mol, in particular 3,000 to 10,000 g / mol, by 1 H-NMR chromatography or by using analytical ultracentrifugation.

[0072] R 1 represents hydrogen or C1-C4 alkyl. Preferably, R 1 represents hydrogen or C1-C3-alkyl, in particular C1-C2-alkyl and very particularly preferably ethyl.

[0073] R 2 Represents OR 11 、-OCO-R 11 、-OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 、-NR 12 -R 14 -CO-NR 13 R 15 、-OR 16 -(O-OC-R 18 ) m and

[0074]

[0075] Preferably, R 2 Indicates -OR 11 、-OCO-R 11 、-OCO-R 14 -CO-OR 11 and-OCO-R 14 -CO-NR 12 R 13 .

[0076] Particularly preferably, R 2 Represents -OCO-R 11 、-OCO-R 14 -CO-OR 11 and-OCO-R 14 -CO-NR12 R 13 .

[0077] Very particularly preferably, R 2 Represents -OCO-R 14 -CO-NR 12 R 13 .

[0078] R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are independently hydrogen, methyl, ethyl, propyl or butyl.

[0079] Preferably, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 independently of one another is hydrogen, methyl or ethyl and in particular only hydrogen.

[0080] R 11 and R 18 It is C6-C 20 Preferably, R 11 and R 18 It is C 8- C 18 Alkyl and especially C 10 -C 14 alkyl.

[0081] R 12 and R 15 and R are independently hydrogen or alkyl. 12 and R 15 It is C 8- C 18 Alkyl and especially C 10 -C 14 alkyl.

[0082] R 13 Indicates C6-C 20 Preferably, R 13 It is C8-C 18 Alkyl and especially C 10 -C 14 alkyl.

[0083] R 14represents an alkylene group, a cycloalkylene group, an arylene group or an aralkylene group. 14 It is C1-C6 alkylene, particularly C2-C4 alkylene, particularly C2 alkylene.

[0084] m is an integer of 1-5, preferably 2 or 3 and in particular 2.

[0085] R 16 is a divalent to hexavalent aliphatic hydrocarbon group. This is derived from a divalent to hexavalent aliphatic alcohol. Preferably, R 16 is a trivalent aliphatic hydrocarbon group. 16 Particularly preferred are those derived from glycerol.

[0086] R 17 is a trivalent bicyclic group. Preferred are groups of the formula

[0087]

[0088] w is an integer in the range of 1 to 5000. w is preferably an integer in the range of 5 to 500 and particularly in the range of 10 to 200.

[0089] x and y are independently of each other an integer in the range of 1 to 5000. Preferably, x and y are independently of each other an integer in the range of 5 to 500 and in particular in the range of 10 to 200.

[0090] z is an integer in the range of 0 to 1000. z is preferably an integer in the range of 0 to 100 and particularly in the range of 0 to 50.

[0091] In each case, the values ​​of x, y and z are chosen so that [ x The mole fraction of the structural unit marked is 10 to 95 mol%, represented by [] y The mole fraction of the structural unit marked is 5 to 90 mol%, and is represented by [] z The molar fraction of the structural units indicated is from 0 to 20 mol %. These percentage data are based in each case on the formula [ x , [] y and[] z The total amount of labeled structural units.

[0092] The copolymer according to the present invention comprises [] x The molar fraction of the structural unit marked is preferably 20 to 90 mol %, and particularly 30 to 70 mol %, based on the formula [] x , [] y and[] z The total amount of the labeled structural units is calculated.

[0093] The copolymer according to the present invention comprises [] yThe molar fraction of the structural unit marked is preferably 10 to 80 mol %, and particularly 30 to 70 mol %, based on the formula [] x , [] y and[] z The total amount of the labeled structural units is calculated.

[0094] The copolymer according to the present invention comprises [] z The molar fraction of the structural unit marked is preferably 0 to 10 mol %, and particularly 0 to 5 mol %, based on the formula [] x , [] y and[] z The total amount of the labeled structural units is calculated.

[0095] Ini is a residue derived from an initiator for cationic polymerization, preferably an organic group. It can be an alkyl group, a cycloalkyl group, an aryl group, an aralkyl group or a heterocyclic group. Alkyl groups and aryl groups are preferred, and C1-C6 alkyl groups are particularly preferred, especially methyl groups.

[0096] Preferred are polymers of formula (I) or (IX), especially polymers of formula (I).

[0097] Preferred are polymers having an Ini group selected from the group consisting of an alkyl group, an aralkyl group or a carboxyalkyl group.

[0098] It is also preferred that R 1 It refers to polymers of C1-C3 alkyl groups, especially methyl or ethyl groups.

[0099] Also preferred is one in which R 2 A polymer selected from the group consisting of:

[0100] -OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 and

[0101]

[0102] Particularly preferably, R 2 The formula is -OCO-R 14 -CO-NR 12 R 13 The group.

[0103] In addition, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9and R 10 Polymers representing hydrogen are preferred.

[0104] It is also preferred that wherein R 11 or R 18 It is C8-C 16 Alkyl polymers.

[0105] It is also preferred that R 12 and R 15 It is C6-C 20 Alkyl, especially C8-C 16 Alkyl polymers.

[0106] Furthermore, it is preferred that R 13 It is C8-C 16 Alkyl polymers.

[0107] It is also preferred that wherein R 14 It is a polymer of C2-C4 alkylene, especially ethylene.

[0108] In further preferred polymers, m is 2 or 3, in particular 2.

[0109] Further preferred polymers include those wherein R 16 These are those polymers derived from the aliphatic hydrocarbon groups of glycerol.

[0110] Also preferred is one in which R 17 It is a polymer of the following group

[0111]

[0112] Very particular preference is given to polymers in which w is an integer in the range 5-500, x and y are independently of one another an integer in the range 5-500, and z is an integer in the range 0-100, with the proviso that [] x The mole fraction of the structural unit marked is 20 to 90 mol%, represented by [] y The mole fraction of the structural unit marked is 10 to 80 mol%, and is represented by [] z The mole fraction of the structural units indicated is from 0 to 20 mol %, in each case based on the formula [] x , [] y and[] z The total amount of the labeled structural units is calculated.

[0113] Highly preferred are polymers of formula (I) wherein R 1 Ethyl, R 2 Represents -OCO-R 14 -CO-NR 12 R 13 , R3 and R 4 Represents hydrogen, R 12 and R 13 Independently represent C8-C 16 Alkyl, R 14 represents a C2-C4 alkylene group, particularly an ethylene group, and w represents an integer in the range of 5-200.

[0114] The polymers according to the invention can be prepared using conventional polymerization methods. Examples are bulk polymerization, solution polymerization or emulsion polymerization or suspension polymerization. These procedures are known to those skilled in the art. Solution polymerization is preferred.

[0115] The oxazolines used for preparing the poly(oxazolines) according to the invention are 2-oxazolines (4,5-dihydrooxazoles) having a C=N double bond between carbon atom 2 and the nitrogen atom. These may be substituted on the 2-, 4- and / or 5-carbon atoms and / or on the 3-nitrogen atom, preferably on the 2-carbon atom and / or on the 3-nitrogen atom.

[0116] Preferably, 2-oxazolines are used which contain a substituent in the 2-position. Examples of such substituents are methyl or ethyl.

[0117] The polymers according to the invention are derived from poly(oxazolines) or poly(oxazines) having selected end groups. These end groups are modified by functionalization. The techniques required for this are known to those skilled in the art.

[0118] Examples of end groups of the poly(oxazoline) or poly(oxazine) starting materials of the polymers according to the invention are halogen atoms, such as fluorine, chlorine, bromine or iodine; or an azide group -N3; ​​or a fluoro(alkyl)-sulfonate group, such as a perfluorobutanesulfonate group -OSO2C4F9, a trifluoromethanesulfonate group -OSO2CF3 or a fluorosulfonate group -OSO2F; or an aryl- or alkylsulfonic acid group such as a tosyl group CH3-C6H4-SO2- or a methanesulfonyl group CH3-SO2-; an unsubstituted, mono- or disubstituted amino group -NH2, -NHR or -NR2 (wherein R = a monovalent organic group), a hydroxyl group -OH, a thiol group -SH, or an ester group -OCOR, a thioester group -SCOR; a phthalimide group or a cyano group -CN and other functional groups obtainable by modification of these end groups.

[0119] The polymers according to the invention can be prepared analogously to methods known from organic chemistry.

[0120] The polymers according to the invention can be prepared by different methods.

[0121] The methods for preparing the polymers of formula (I), (II), (IX) or (X) are common to the preparation of poly(oxazoline) or poly(oxazine) by cationic ring-opening polymerization of oxazoline or oxazine. The polymerization is preferably carried out in solution and in the presence of an initiator. Examples of initiators are electrophilic reagents, such as salts or esters of aromatic sulfonic acids or carboxylic acids, or salts or esters of aliphatic sulfonic acids or carboxylic acids, or aromatic halogen compounds. Examples of preferred initiators are esters of aryl sulfonic acids, such as methyl toluenesulfonate, esters of alkanesulfonic acids such as trifluoromethanesulfonic acid, or mono- or dibromobenzene.

[0122] Polar aprotic solvents are generally used as solvents, such as acetonitrile, dimethylformamide, dimethylacetamide, ethylene carbonate or dimethyl sulfone.

[0123] The reaction temperature is generally between 20 and 180°C, in particular in the range of 70 to 130°C.

[0124] The reaction duration of the polymerization is generally between 5 minutes and 24 hours.

[0125] The hydrolysis of the poly(oxazoline) or poly(oxazine) is preferably carried out in solution, in particular in an aqueous solution or an alcohol-water solution. An inorganic or organic acid can be used as the acid. Preferably, an inorganic acid or a carboxylic acid is used. Examples thereof are hydrochloric acid, sulfuric acid, nitric acid, acetic acid or formic acid, preferably acetic acid. Suitable bases include, for example, alkali metal hydroxides, such as sodium hydroxide or potassium hydroxide.

[0126] The reaction temperature of the hydrolysis is generally between 20 and 120°C, in particular in the range of 30-80°C.

[0127] The reaction duration of the hydrolysis is generally between 5 minutes and 24 hours.

[0128] The preparation of “degPAOx” as starting material for producing the second group of polymers according to the invention is described, for example, in WO 2022 / 106049 A1.

[0129] Preference is given to processes in which the poly(oxazoline) or poly(oxazine) used is obtained by hydrolysis, in particular by acidic hydrolysis.

[0130] The end groups of the poly(oxazolines) or poly(oxazines) used as starting materials may be further modified prior to further processing.

[0131] For example, a polymer having carboxylate end groups can be converted to a polymer having hydroxyl end groups. This can be accomplished by saponification in aqueous or aqueous-alcoholic solution in the presence of a strong base, such as an alkali metal alkoxide, such as sodium methoxide.

[0132] The reaction temperature of the saponification is usually between 10 and 120°C, particularly in the range of 20-60°C.

[0133] The reaction duration of the saponification is generally between 1 and 24 hours.

[0134] The polymer with hydroxyl or amino end groups can be further modified by reaction with dicarboxylic anhydrides. This produces a polymer with carboxyl end groups. For example, a polymer with hydroxyl end groups can be converted into a polymer with end groups derived from dicarboxylic acids, such as in the reaction of a hydroxyl-terminated polymer with an anhydride of an aliphatic dicarboxylic acid such as succinic anhydride. The reaction can be carried out in an aprotic solvent such as dimethylformamide in the presence of a tertiary amine such as dimethylaminopyridine and triethylamine.

[0135] The reaction temperature in this reaction is generally between 10 and 120°C, in particular in the range of 20-60°C.

[0136] The reaction duration of this reaction is generally between 1 and 24 hours.

[0137] Finally, the polymer with the end group derived from dicarboxylic acid can be further modified by reacting with primary amine or secondary amine. This produces a polymer with an amide end group. For example, the carboxyl end group of the polymer with the end group derived from dicarboxylic acid can be converted into the corresponding carboxylic acid amide by reacting with primary amine or secondary amine. The reaction can be carried out in a polar aprotic solvent, such as chloroform, in the presence of a tertiary amine, such as dimethylaminopyridine. The reaction is also carried out in the presence of known coupling reagents, such as N-hydroxysuccinimide ester (NHS ester), N-hydroxysuccinimide (NHS), dicyclohexylcarbodiimide ester (DCC ester) or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide ester (EDC ester) or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC-HCl).

[0138] The reaction temperature of the reaction is usually between 10 and 120°C, in particular in the range of 20-60°C.

[0139] The reaction duration of this reaction is generally between 1 and 24 hours.

[0140] In the preparation of the polymers according to the invention, the radical R 2 In the preparation of the polymers according to the invention having degPAOx groups, further variations of the preparation methods can also be used.

[0141] The polymer of formula (I) or (II) can be prepared by a process comprising the following steps, wherein R 2 Indicates -OR 11 or-OCO-R 11 :

[0142] a) by cationic polymerization of 2-oxazoline or 2-oxazine in the presence of a cationic polymerization initiator to provide a polymer of formula (Ia) or (IIa)

[0143]

[0144] b) Termination of cationic polymerization by adding a compound of formula (XI), (XII) or (XIIa)

[0145] R 11 -OH(XI),R 11 -COOH(XII),

[0146] R 11 -CO-O-OC-R 11 (XIIa),

[0147] Among them, in these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 11 and w have the meanings defined above, i is an integer of 1 to 4 and An is an i-valent anion.

[0148] A polymer of formula (I) or (II), wherein R 2 express

[0149]

[0150] It can be prepared by a method comprising the following steps:

[0151] a) by cationic polymerization of 2-oxazoline or 2-oxazine in the presence of a cationic polymerization initiator to provide a polymer of formula (Ia) or (IIa)

[0152]

[0153] Among them, Ini, R 1 , R 3 , R 4 , R 5 and w have the meanings defined above, i is an integer of 1 to 4 and An is an i-valent anion,

[0154] c) terminating the cationic polymerization by adding an inorganic or organic azide, thereby forming an azide-terminated polymer of formula (Ib) or (IIb),

[0155]

[0156] d) reacting the azide-terminated polymer of formula (Ib) or (IIb) obtained from step c) with a cycloalkyne of formula (XIII) to produce an ester-terminated polymer of formula (Ic) or (IIc)

[0157]

[0158]

[0159] Where R =

[0160] and

[0161] e) reacting an ester-terminated polymer of formula (Ic) or (IIc) with an amine of formula (XIV)

[0162] R 12 R 13 NH(XV)

[0163] To produce a polymer of formula (I) or (II), wherein R 2 has the meaning defined above,

[0164] Among them, in these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 12 , R 13 and w have the meanings defined above, i is an integer of 1 to 4 and An is an i-valent anion.

[0165] Where R 2 Represents -OCO-R 14 -CO-OR 11 or-OCO-R 14 -CO-NR 12 R 13 The polymer of formula (I) or (II) can be prepared by a process comprising the following steps:

[0166] a) by cationic polymerization of 2-oxazoline or 2-oxazine in the presence of a cationic polymerization initiator to provide a polymer of formula (Ia) or (IIa)

[0167]

[0168] f) terminating the cationic polymerization by adding a compound of formula (XII) or (XIIa) to give a hydroxyl-terminated polymer of formula (Id) or (IId)

[0169]

[0170] g) reacting a polymer of formula (Id) or (IId) with an anhydride of formula (XVI) to produce a polymer of formula (Ie) or (IIe)

[0171]

[0172]

[0173] and

[0174] h) reacting the polymer of formula (Ie) or (IIe) from step g) with an alcohol of formula (XI) or with an amine of formula (XV) to produce a polymer of formula (I) or (II), wherein R 2 has the meaning defined above,

[0175] R 11 -OH(XI), R 12 R 13 NH(XV)

[0176] Among them, in these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 11 , R 12 , R 13 , R 14 and w have the meanings defined above, i is an integer of 1 to 4 and An is an i-valent anion.

[0177] The polymer of formula (I) or (II) can be prepared by a process comprising the following steps, wherein R 2 Indicates -OR 16 -(O-OC-R 18 ) m :

[0178] a) by cationic polymerization of 2-oxazoline or 2-oxazine in the presence of a cationic polymerization initiator to provide a polymer of formula (Ia) or (IIa)

[0179]

[0180] i) terminating the cationic polymerization by adding a carboxylic acid, preferably acetic acid, to result in a carboxyl-terminated polymer of formula (If) or (IIf)

[0181]

[0182] j) reacting the polymer of formula (If) or (IIf) from step i) with an alkali metal alkoxide, preferably sodium methoxide, to produce a polymer of formula (Ig) or (IIg)

[0183]

[0184] k) reacting the polymer of formula (Ig) or (IIg) from step j) with a dicarboxylic anhydride of formula (XVI), preferably succinic anhydride, to produce a polymer of formula (Ih) or (IIh)

[0185]

[0186] and

[0187] l) reacting the polymer of formula (Ih) or (IIh) from step k) with an alcohol of formula (XVII) to produce a polymer of formula (I) or (II), wherein R 2 has the meaning defined above,

[0188] HOR 16 -(O-OC-R 18 ) m (XVII)

[0189] Among them, in these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 14 , R 16 , R 18 , m and w have the meanings as defined above, i is an integer of 1 to 4 and An is an i-valent anion.

[0190] The polymer of formula (I) or (II) can be prepared by a process comprising the following steps, wherein R 2 Indicates -NR 12 -R 14 -CO-NR 13 R 15 :

[0191] a) by cationic polymerization of 2-oxazoline or 2-oxazine in the presence of a cationic polymerization initiator to provide a polymer of formula (Ia) or (IIa)

[0192]

[0193] m) terminating the cationic polymerization by adding an alkali metal phthalimide to produce an N-phthalimide terminated polymer,

[0194] n) reacting the polymer from step m) with hydrazine to produce a polymer of formula (Ii) or (IIi)

[0195]

[0196]

[0197] o) reacting the polymer of formula (Ii) or (IIi) from step n) with a dicarboxylic anhydride of formula (XVI), preferably succinic anhydride, to produce a polymer of formula (Ij) or (IIj)

[0198]

[0199] and

[0200] p) reacting the polymer of formula (Ij) or (IIj) from step o) with an amine of formula (XV) to produce a polymer of formula (I) or (II), wherein R 2 has the meaning defined above,

[0201] R 12 R 13 NH(XV),

[0202] Among them, in these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 12 , R 13 , R 14 and w have the meanings defined above, i is an integer of 1 to 4 and An is an i-valent anion.

[0203] In the preparation of the copolymers of the second polymer group according to the invention, i.e. the copolymers containing degPAOx groups, i.e. structural units of the formula (III), (IV) and optionally (V) or of the formula (VI), (VII) and optionally (VIII), different starting materials can be used.

[0204] These copolymers may be linear or branched.

[0205] The linear type is a copolymer of formula (IX) or (X). These can be prepared analogously to the first group of linear polymers, ie polymers of formula (I) or (II). For this purpose, the radical R 2 Functionalized polyoxazoline or polyoxazine are fully or partially hydrolyzed.Then the multipolymer obtained is oxidized, and acylated again in the case of complete hydrolysis, which directly leads to the multipolymer according to the second polymer group of the present invention. The details of the preparation of the multipolymer of formula (IX) and (X) are further provided below.

[0206] By using the group R 2Partial oxidation of the functionalized polyalkylene imine, thereby partially oxidizing the branched type of the copolymer containing degPAOx, and the resulting product can be functionalized, for example, by reaction with an activated acyl derivative, such as an activated ester, or with an acyl halide, to produce copolymers of the second polymer group. Details of the preparation of these copolymers are further given below. Commercially available polyethyleneimines generally have a branched structure; therefore, the polymers derived therefrom are also branched.

[0207] These methods for preparing degPAOx copolymers are disclosed in WO 2022 / 106049 A1.

[0208] The linear polymer of formula (IX) or (X) can be prepared by a process comprising the following steps, wherein R 2 Indicates -OR 11 or-OCO-R 11 :

[0209] q) providing a polymer of formula (I) or (II), wherein R 2 Indicates -OR 11 or-OCO-R 11 ,

[0210] r) partial hydrolysis of the polymer of formula (I) or (II) obtained in step q) to produce a copolymer of formula (Ik) or (IIk)

[0211]

[0212] s) reacting the copolymer of formula (Ik) or (IIk) from step r) with an oxidizing agent, thereby obtaining a copolymer of formula (IX) or formula (X), wherein R 2 has the meaning defined above,

[0213] In these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , w, x, y and z have the meanings defined above, and wherein za is an integer with value wx.

[0214] A linear polymer of formula (IX) or (X), wherein R 2 express

[0215]

[0216] It can be prepared by a method comprising the following steps:

[0217] t) providing a polymer of formula (I) or (II), wherein R 2 express

[0218]

[0219] u) partial hydrolysis of the polymer of formula (I) or (II) obtained in step t) to produce a copolymer of formula (II) or (IIl)

[0220]

[0221] v) reacting the copolymer of formula (Il) or (IIl) from step u) with an oxidizing agent, thereby obtaining a copolymer of formula (IX) or formula (X), wherein R 2 has the meaning defined above,

[0222] In these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 , R 10 , R 12 , R 13 , w, x, y and z have the meanings defined above, and wherein za is an integer with value wx.

[0223] The linear polymer of formula (IX) or (X) can be prepared by a process comprising the following steps, wherein R 2 Represents -OCO-R 14 -CO-OR 11 or-OCO-R 14 -CO-NR 12 R 13 :

[0224] w) providing a polymer of formula (I) or (II), wherein R 2 OCO-R 14 -CO-OR 11 or-OCO-R 14 -CO-NR 12 R 13 ,

[0225] x) partial hydrolysis of the polymer of formula (I) or (II) obtained in step wo) to produce a copolymer of formula (Im) or (IIm)

[0226]

[0227] y) reacting the copolymer of formula (Im) or (IIm) from step x) with an oxidizing agent, thereby obtaining a copolymer of formula (IX) or formula (X), wherein R 2 has the meaning defined above,

[0228] In these formulas, Ini, R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , w, x, y and z have the meanings defined above, and wherein za is an integer with value wx.

[0229] The second polymer group is a branched copolymer, wherein R 2 Indicates -OR 11 、-OCO-R 11 、-OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 、-NR 12 -R 14 -CO-NR 13 R 15 ,

[0230] -OR 16 -(O-OC-R 18 ) m or

[0231]

[0232] It can be prepared by a method comprising the following steps:

[0233] z) providing a branched polyethyleneimine or polypropyleneimine, at least one end group of which is substituted with a group R 2 Functionalization,

[0234] aa) partial oxidation of the functionalized polymer obtained in step z), and

[0235] bb) by reacting with an acyl halide R 1 -CO-Hal reaction converts -CO-R1 groups are introduced into the polymer obtained in step aa) to produce a branched copolymer containing structural units of the formulae (III), (IV) and optionally (V) or containing structural units of the formulae (VI), (VII) and optionally (VIII),

[0236] In these formulas, R 1 , R 2 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 and m have the meanings defined above.

[0237] The oxidation in steps s), v), y) and aa) is preferably carried out in solution, in particular in an aqueous or alcohol-aqueous solution. Oxidants known per se can be used as oxidizing agents. Examples thereof are peroxy compounds, hypochlorites, chlorine or oxygen, in particular hydrogen peroxide.

[0238] Peroxy compounds are preferably used. Examples of these are hydrogen peroxide, peracids, organic peroxides or organic hydroperoxides, in particular hydrogen peroxide.

[0239] The process wherein the oxidizing agent used is hydrogen peroxide is preferred.

[0240] The amount of oxidizing agent is selected so that the desired proportion of oxidized structural units is formed in the polymer backbone.

[0241] The reaction temperature in this reaction is usually between 10 and 80°C, particularly in the range of 20 to 40°C.

[0242] The reaction duration of the oxidation is usually between 5 minutes and 5 days.

[0243] The polymers according to the invention can be used for the preparation of formulations containing pharmaceutical active ingredients or agrochemical active ingredients.

[0244] Preferably, the polymers according to the invention are used for the preparation of formulations containing pharmaceutically active ingredients or agrochemically active ingredients. These are in particular formulations containing vaccines or nucleic acids, for example RNA or DNA.

[0245] Due to their good surfactant effect, biocompatibility and stealth effect, the polymers according to the invention are outstandingly suitable for use in the field of active ingredient administration. These uses are also subject matter of the present invention.

[0246] In particular, the polymers of the second group are preferably suitable for the preparation of formulations containing pharmaceutical or agrochemical active ingredients due to their biodegradability.

[0247] The polymers according to the invention can be used as lipids due to their amphiphilic nature.They can be present dispersed in a hydrophilic liquid, for example as an emulsion or suspension.

[0248] Preferably, the polymer according to the invention is present in the form of particles, in particular in the form of nanoparticles, in a hydrophilic liquid, for example water or a water-alcohol mixture.

[0249] The present invention therefore also relates to particles, in particular nanoparticles, comprising the polymers described above.

[0250] The average diameter D is preferably 50 Nanoparticles smaller than 1 μm, preferably 20 to 500 nm.

[0251] Granules which contain one or more pharmaceutically active ingredients or agrochemically active ingredients are very particularly preferred.

[0252] Particularly preferred granules contain, in addition to the polymers according to the invention, at least one pharmaceutically active ingredient and suitable excipients and additives.

[0253] Preferably, the particles form a dispersed phase in a liquid containing water and / or a water-miscible compound.

[0254] The proportion of particles in the dispersion can cover a wide range. Typically, the proportion of particles in the dispersion medium is 0.5 to 20% by weight, preferably 1 to 5% by weight.

[0255] The particles can be prepared by precipitation, preferably by nanoprecipitation. For this purpose, the polymer according to the invention, which is less hydrophilic or non-hydrophilic due to the presence of hydrophobic groups, is dissolved in a water-miscible solvent, for example acetone. The solution is added dropwise to the hydrophilic dispersion medium. This is preferably done with vigorous stirring. This facilitates the preparation of smaller particles. The polymer is isolated in finely divided form in the dispersion medium.

[0256] Alternatively, the particles can also be prepared by emulsification, preferably by nanoemulsion. For this purpose, the polymer according to the invention, which is low hydrophilic or non-hydrophilic due to the presence of hydrophobic groups, is dissolved in a water-immiscible solvent, for example dichloromethane or ethyl acetate. This solution is combined with a hydrophilic dispersion medium, thereby preferably resulting in the formation of two liquid phases. This mixture is then emulsified by inputting energy, preferably by ultrasonication with ultrasound.

[0257] In addition to the polymer according to the invention, one or more active ingredients and / or one or more excipients and additives may be present when the polymer is dispersed in the dispersion medium. Alternatively, these active ingredients and / or excipients and additives may be added after the polymer has been dispersed in the hydrophilic liquid.

[0258] Microfluidics for the preparation of lipid nanoparticles (“LNPs”) is particularly suitable as a formulation method here.

[0259] For example, LNP can be prepared using a microfluidic device by an ethanol dilution method. For this purpose, a lipid solution prepared in ethanol and active ingredients, such as RNA, are dissolved in a suitable buffer solution. In order for active ingredients to be encapsulated in LNP, cationic lipids or pH-sensitive cationic lipids are used for lipid components. Lipid solution and buffered active ingredient solution are introduced into a microfluidic device, where for example positively charged lipids and negatively charged RNA form complexes via electrostatic interactions. Cationic RNA-lipid complexes are then merged with other lipids to form LNP. The example of other lipids is cholesterol, phospholipids, PEG lipids or PAOx lipids.

[0260] The polymer particles can be separated from the hydrophilic liquid in different ways. Examples include centrifugation, ultrafiltration or dialysis.

[0261] The polymer dispersion can be further purified after preparation. Common methods include purification by dialysis, ultrafiltration, filtration or centrifugation.

[0262] The following examples illustrate the invention without limiting it.

[0263] The synthesis of poly(2-ethyl-2-oxazoline) lipid is described below. BRIEF DESCRIPTION OF THE DRAWINGS

[0264] Figure 1 Schematic diagram showing the synthesis of PEtOx lipids. DETAILED DESCRIPTION

[0265] The following abbreviations are used in the examples:

[0266] MeOH: Methanol

[0267] EtOH: ethanol

[0268] NaOMe: sodium methoxide

[0269] DMAP: dimethylaminopyridine

[0270] DMF: dimethylformamide

[0271] NHS: N-hydroxysuccinimide

[0272] EDC-HCl: 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride

[0273] Material

[0274] All chemicals and solvents were purchased from commercial suppliers and used without further purification unless otherwise stated. 2-Ethyl-2-oxazoline (EtOx, 99+%), triethylamine (NEt3, 99.7%), and EtOx were purchased from Sigma Aldrich. 2-Ethyl-2-oxazoline was dried over calcium hydride and distilled under an argon atmosphere. Methyl toluenesulfonate (MeOTs, 98%), methyl toluenesulfonate was dried over barium oxide and distilled under an argon atmosphere. Hydrochloric acid (37%) was purchased from Fisher Chemicals. Aqueous hydrogen peroxide (30% w / w) was obtained from Carl Roth. Acetyl chloride (approximately 90%) was purchased from Merck Schuchardt. Amberlite IRA-67 was obtained from Merck and washed several times with deionized water before use. N,N-dimethylformamide (DMF) and acetonitrile were dried in a solvent purification system (MB-SPS-800 from M Braun). Phosphate buffered saline (PBS) was obtained from Biowest. Succinic anhydride (≥99%), N-hydroxysuccinimide (NHS, ≥99%), sodium methoxide (0.5 M in methanol), and BCN-NHS were obtained from Sigma Aldrich. Ditetradecylamine (95%) was purchased from AmBeed. N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC-HCL) was obtained from Apollo Scientific. Triethylamine (>99%) and 4-dimethylaminopyridine (DMAP, >99%) were obtained from TCI.

[0275] Implementation of measurement

[0276] Protons were measured on a Bruker AC 300 MHz or Bruker AC 400 MHz spectrometer. 1H) Nuclear magnetic resonance (NMR) spectra. Correlation spectrum (COSY) NMR, heteronuclear single quantum correlation spectrum (HSQC) NMR, heteronuclear multiple bond correlation (HMBC) NMR spectra and DOSY NMR spectra were recorded on a Bruker AC 400MHz spectrometer. Measurements were performed using D2O, d4-methanol or deuterated chloroform as solvents at room temperature. Chemical shifts (δ) were reported in parts per million (ppm) relative to the remaining undeuterated solvent resonance signal. Infrared spectroscopy (IR) was performed on a Shimadzu IRAffinity-1CE system equipped with a QuestATR type single reflection ATR cuvette with a diamond crystal for measuring over an extended range.

[0277] Size Exclusion Chromatography (SEC) was performed using two different setups. A 1000 nm HPLC-MS / MS instrument was used with a PSS degasser, G1310A pump, G1329A autosampler, Techlab oven, G1362A refractive index detector (RID) and PSS GRAM-protected / 30 / The measurements in N,N-dimethylacetamide (DMAc) were performed on an Agilent 1200 series system with a column (10 μm particle size). DMAc with 0.21 wt % LiCl was used as the eluent. The flow rate was 1 mL min -1 And the oven temperature is 40°C. 400-1,000,000g mol -1 Polystyrene (PS) or polymethyl methacrylate (PMMA) standards were used for the calculation of molar mass. The Shimadzu system (Shimadzu Corp., Kyoto, Japan) equipped with SCL-10A VP system controller, SIL-10AD VP autosampler, LC-10AD VP pump, RID-10A RI detector, CTO-10A VP oven and PSSSDV protection / lin S-column (5 mm particle size) was used for the measurement in chloroform. A mixture of chloroform / isopropanol / -triethylamine (94 / 2 / 4 volume %) was used as eluent. The flow rate was 1 mL min -1 And the oven temperature is 40°C. 400-100,000g mol -1 PS standards of 100 μg / ml were used to calibrate the system.

[0278] pass 1 Characterization of polymers by H-NMR spectroscopy

[0279] The first step in the preparation of PEtOx lipids is to synthesize PEtOx with various repeat units (20, 40, 50, 60 and 100) by CROP (see Synthesis of PEtOx-OAc). CROP is terminated by adding acetic acid. 1H-NMR spectroscopy was used to determine the degree of polymerization via the conversion of monomers into polymers. The hydrolysis was carried out under alkaline conditions (see Synthesis of POx-OH). To obtain complete hydrolysis, NaOMe was used to react overnight. 1 The successful synthesis was confirmed by H-NMR, which clearly showed the disappearance of the signal of the CH3 group attributable to the OAc-ω-terminal group of PEtOx-OAc. In order to extend the linker unit, PEtOx-OH was reacted with succinic anhydride (see Synthesis of Succinylated PEtOx). The product was also prepared by 1 H NMR spectroscopy. To introduce lipid groups, the end groups of succinylated PEtOx were activated by EDC coupling and NHS and reacted with ditetradecylamine. The product was obtained by 1 H-NMR Characterization.

[0280] Cationic Ring-Opening Polymerization of Ethyl Oxazoline (PEtOx OAc)

[0281] Synthesized according to M. Dirauf, A. Erlebach, C. Weber, S. Hoeppener, JR Buchheim, M. Sierka, US Schubert, Macromolecules 2020, 53, 3580-3590.

[0282] Methyl toluenesulfonate (1 eq.) and ethyl oxazoline (20, 40, 50, 60, 100 eq., depending on the desired chain length) were dissolved in anhydrous acetonitrile and heated under reflux. The mixture was cooled, acetic acid (1.5 eq.) and triethylamine (2 eq.) were added successively and stirred at 50° C. overnight. The reaction mixture was diluted with CHCl (100 mL), washed with saturated NaHCO solution (3×200 mL) and brine. The combined organic phases were dried over Na2SO4, the volatile fractions were removed under reduced pressure and dried in vacuo at 40°C overnight.

[0283] oeLh 50 -OAc: 1 H NMR (300 MHz, CDCl3): δ 0.99-1.23 (br, 150H, CH2-CH3); 2.01-2.13 (br, 3H, CO-CH3); 2.18-2.56 (m, 100H, CH2-CH3); 2.98-3.13 (br, 3H, CH3-N), 3.30-3.66 (br, 200H, N-CH2-CH2, main chain) ppm.

[0284] SEC (DMAc + 0.21 wt% LiCl): M n=8820 g mol -1 ,

[0285] Synthesis of PEtOx-OH

[0286] The synthesis was also carried out according to M. Dirauf, A. Erlebach, C. Weber, S. Hoeppener, JR Buchheim, M. Sierka, US Schubert, Macromolecules 2020, 53, 3580-3590.

[0287] PEtOx-OAc (1 equiv.) was dissolved in anhydrous MeOH (0.15 g mL -1 ) and NaOMe (0.1 eq., 0.5 M in MeOH) was added under vigorous stirring. The reaction mixture was stirred at room temperature overnight and then the solvent was removed under reduced pressure. The residue was dissolved in CHCl3 and washed with saturated NaHCO3 (3×200 mL) and brine (200 mL). The combined organic phases were dried over Na2SO4 and the solvent was removed under reduced pressure. The product was then dissolved in CH2Cl2 and precipitated in ice-cold diethyl ether. The polymer was dried overnight at 40°C in a vacuum.

[0288] oeLh 50 -OH: 1 H NMR (300 MHz, CDCl3): δ 0.99-1.25 (br, 150H, CH2-CH3); 2.19-2.59 (m, 100H, CH2-CH3); 3.00-3.12 (br, 3H, CH3-N), 3.31-3.69 (br, 200H, N-CH2-CH2, main chain) ppm.

[0289] SEC (DMAc + 0.21 wt% LiCl): M n =9240 g mol -1 ,

[0290] Synthesis of succinylated PEtOx

[0291] PEtOx-OH (1 equivalent) and DMAP (1.05 equivalents) are dissolved in anhydrous DMF (c=0.145M). Triethylamine (0.1 equivalent) and succinic anhydride (3 equivalents) are added to the mixture and stirred overnight at room temperature. The reaction mixture is then precipitated in ice-cold diethyl ether and then dissolved in dichloromethane. The mixture is then washed with saturated NH4Cl solution and the combined organic phase is dried over MgSO4. Volatile components are removed under reduced pressure and dissolved again in dichloromethane and precipitated in ice-cold diethyl ether. The resulting solid is dried overnight in a vacuum.

[0292] oeLh 50 -COOH: 1 H NMR (300MHz, CDCl3): δ0.96-1.13(br,150H,CH2-CH3); 2.13-2.43(m,100H,CH2-CH3); 2.44-2.6 7(br,4H,CO-CH2-CH2-CO); 2.95-3.01(br,3H,CH3-N),3.28-3.58(br,200H,N-CH2-CH2, main chain)ppm.

[0293] SEC (DMAc + 0.21 wt% LiCl): M n =8900 g mol -1 ,

[0294] Synthesis of Succinylated PEtOx and Ditetradecylamine

[0295] Succinylated PEtOx (1 equivalent) was dissolved in anhydrous CHCl and DMAP (0.1 equivalent), NHS (2.5 equivalents) and EDC-HCl (3 equivalents) were added and stirred at room temperature for 3h. Ditetradecylamine (4 equivalents) was added and the mixture was stirred at 45°C overnight. The mixture was then precipitated in ice-cold diethyl ether, dissolved in CH2Cl2 again and cooled to -20°C for 3 to 5h. The precipitate was filtered (0.25 μm PTFE filter), precipitated in ice-cold diethyl ether and dialyzed with EtOH: water (1:1, 1000Da MWCO dialysis membrane) for 3 days, dialyzed with water for 2 days and then lyophilized.

[0296] oeLh 50 -Lipids: 1H NMR (300MHz,CDCl3):δ0.81(t,6H,lipid CH2-CH3);0.97-1.12(br,150H,CH2-CH3);1.13-1.27(br,36H,lipid alkyl chain);1.38-1.45(br,2H,N-CH2-CH2);1.45-1.57(br,2H,N-CH2-CH2);2.14 -2.42 (m, 100H, CH2-CH3); 2.51-2.61 (m, 4H, CO-CH2-CH2-CO); 2.94-3.02 (br, 3H, CH3-N), 3.15 (t, 2H, N-CH2); 3.17-3.23 (m, 2H, N-CH2); 3.28-3.58 (br, 200H, N-CH2-CH2, main chain) ppm.

[0297] SEC (DMAc + 0.21 wt% LiCl): M n =9300 g mol -1 ,

[0298] Figure 2 Schematic diagram showing the synthesis of degPOx lipids. Here, the linker is activated by strain-promoted azide-alkyne cycloaddition (SPAAC) and then the lipid is coupled to the linker.

Claims

1. Polymers of formula (I) or (II) Or a polymer containing, based on all structural units, 10 to 95 mol % of structural units of formula (III), 5 to 90 mol % of structural units of formula (IV) and 0 to 20 mol % of structural units of formula (V) or a compound containing 10 to 95 mol % of a structural unit of formula (VI), 5 to 90 mol % of a structural unit of formula (VII) and 0 to 20 mol % of a structural unit of formula (VIII) Which has the structural part -CHR 4 -、-CHR 5 - or CHR 7 - or having the structural part -CHR 5 -、-CHR 7 -or-CHR 10 - at least one of the structural units of formula (III), (IV) or (V) or formula (VI), (VII) or (VIII) with the group R 2 Covalent bonding, in Ini is a group derived from a cationic polymerization initiator, R 1 Selected from the group consisting of: hydrogen or C1-C4 alkyl, R 2 Selected from the group consisting of: -OR 11 、-OCO-R 11 、-OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 、-NR 12 -R 14 -CO-NR 13 R 15 、-OR 16 -(O-OC-R 18 ) m and R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are independently hydrogen, methyl, ethyl, propyl or butyl, R 11 It is C6-C 20 alkyl, R 12 and R 15 are independently hydrogen or alkyl, R 13 It is C6-C 20 alkyl, R 14 is an alkylene group, a cycloalkylene group, an arylene group or an aralkylene group, R 16 is an m+1-valent aliphatic hydrocarbon group, R 18 It is C6-C 20 Alkyl, provided that the group R 16 Several groups R 18 can be different within a given definition, m is an integer from 1 to 5, R 17 is a trivalent bicyclic radical, and w is an integer in the range of 1-5000.

2. The polymer according to claim 1, wherein the polymer has the structure of formula (IX) or (X) Where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 has the meaning defined in claim 1, x and y independently represent integers in the range of 1 to 5000, z is an integer in the range 0-1000, The condition is given by [] x The mole fraction of the structural unit marked is 10 to 95 mol%, represented by [] y The mole fraction of the structural unit marked is 5 to 90 mol%, and is represented by [] z The mole fraction of the structural units indicated is from 0 to 20 mol %, in each case based on the formula [] x , [] y and[] z The total amount of labeled structural units.

3. The polymer according to claim 1 or 2, wherein the polymers are of formula (I) or (IX).

4. The polymer according to claim 1, wherein the polymers are of formula (I).

5. The polymer according to at least one of claims 1 to 4, wherein Ini is selected from the group consisting of alkyl, aralkyl or carboxyalkyl.

6. The polymer according to claim 1, wherein R 1 represents a C1-C3 alkyl group, preferably a methyl group or an ethyl group.

7. The polymer according to claim 1, wherein R 2 Selected from the group consisting of: -OCO-R 14 -CO-OR 11 、-OCO-R 14 -CO-NR 12 R 13 and In particular, the following radicals: -OCO-R 14 -CO-NR 12 R 13 。 8. The polymer according to claim 1, wherein R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 Represents hydrogen.

9. The polymer according to claim 1, wherein R 11 It is C8-C 16 alkyl.

10. The polymer according to claim 1, wherein R 12 and R 15 It is C6-C 20 Alkyl, especially C8-C 16 alkyl.

11. The polymer according to claim 1, wherein R 13 It is C8-C 16 alkyl.

12. The polymer according to claim 1, wherein R 14 It represents a C2-C4 alkylene group, preferably an ethylene group. 13 . The polymer according to claim 1 , wherein m is 2 or 3, preferably 2.

14. The polymer according to at least one of claims 1 to 13, wherein R 16 It is an aliphatic hydrocarbon group derived from glycerol.

15. The polymer according to at least one of claims 1 to 14, wherein R 17 Is a group of the formula 16. The polymer according to claim 1, wherein w represents an integer in the range of 5-500, x and y are independently an integer in the range of 5-500, z is an integer in the range of 0-100, provided that [] x The mole fraction of the structural unit marked is 20 to 90 mol%, represented by [] y The mole fraction of the structural unit marked is 10 to 80 mol%, and is represented by [] z The mole fraction of the structural units indicated is from 0 to 20 mol %, in each case based on the formula [] x , [] y and[] z The total amount of labeled structural units.

17. The polymer according to claim 4, wherein R 1 is ethyl, R 2 Represents -OCO-R 14 -CO-NR 12 R 13 , R 3 and R 4 Represents hydrogen, R 12 and R 13 Independently represent C8-C 16 Alkyl, R 14 represents a C2-C4 alkylene group, particularly an ethylene group, and w represents an integer in the range of 5-200.

18. Use of a polymer according to at least one of claims 1 to 17 for preparing a formulation containing pharmaceutically active ingredients or agrochemically active ingredients.

19. The use according to claim 18, wherein the active ingredient is a pharmaceutical active ingredient or an agrochemical active ingredient, in particular a vaccine or a nucleic acid.

20. Particles, in particular nanoparticles, comprising a polymer according to at least one of claims 1 to 17.

Images