CN113354811B

CN113354811B - Oxazoline polymer with side chain containing amino and/or hydroxyl, preparation method and application thereof

Info

Publication number: CN113354811B
Application number: CN202010103178.4A
Authority: CN
Inventors: 刘润辉; 周敏
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2020-02-19
Filing date: 2020-02-19
Publication date: 2024-05-07
Anticipated expiration: 2040-02-19
Also published as: CN113354811A

Abstract

The invention relates to an oxazoline polymer with an amino group and/or hydroxyl group in a side chain, a preparation method and application thereof. The oxazoline polymer with the side chain containing the amino group can be used in the fields of antibacterial, antifungal, antiviral, anti-mite, antitumor, cell adhesion, tissue engineering, drug modification, protein protection, drug synergy, drug delivery, gene delivery, self-assembly materials and the like; the oxazoline polymer with the side chain containing hydroxyl can be used in the fields of surface antifouling, protein modification and protection, cell protection, tissue and organ freeze protection, drug modification and the like; the oxazoline polymer with the side chain containing the amino group and the hydroxyl group has the functions of the oxazoline polymer with the side chain containing the amino group and the hydroxyl group.

Description

Oxazoline polymer with side chain containing amino and/or hydroxyl, preparation method and application thereof

Technical Field

The invention belongs to the technical field of polymers, and particularly relates to an oxazoline polymer with amino groups or/and hydroxyl groups in side chains, and a preparation method and application thereof.

Background

The polypeptide polymer has a secondary structure and biocompatibility similar to those of natural proteins, so that the polypeptide polymer has wide application prospects in the field of biomedical materials, such as the fields of protein simulation, antibacterial materials, drug and gene transfer, stimulus response polypeptides, tissue engineering and the like. However, since polypeptide polymers are easily degraded by proteases to lose their original functions, the study of protease hydrolysis resistant polypeptide mimics is of great importance. Since the 1960 s, 2-substituted oxazoline polymers have been found, and have been widely studied in many fields, for example, fields of thermally responsive materials, antifouling materials, drug delivery, nano-administration, and drug and protein coupling, as a substance that mimics the function of PEG due to their good biocompatibility.

Although poly-2-methyl oxazoline and poly-2-ethyl oxazoline and derived materials thereof have been reported, only few documents report the synthesis of oxazoline polymers with amino groups in side chains and very limited functions thereof in the biomedical field, for example, no documents have disclosed that the polymers can have antibacterial functions, antitumor functions and the like. One of the important reasons is that the conditions for the polymerization of oxazoline monomers are generally mainly as follows: the polymerization reaction is carried out by taking methyl p-toluenesulfonate and methyl trifluoromethanesulfonate as initiators and acetonitrile as a solvent. And the group of the Rainer Jordan problem in 2006 reported that oxazoline polymers with pendant amino groups could not be successfully prepared under normal conditions, (Macromol Chem Physics 2006,207 (2): 183-192.). In addition, polymers having hydroxyl groups in the side chains have been shown to have important functions in many fields of research.

Therefore, there is an urgent need in the art to develop oxazoline polymers having amino groups in side chains, which have functional and application values in the biomedical field.

Disclosure of Invention

The invention aims to provide oxazoline polymers with side chains containing amino groups and/or hydroxyl groups, a preparation method thereof and application thereof in the field of biomedical materials.

In a first aspect of the invention, there is provided an oxazoline polymer comprising one or more of the repeat units of formula E ₁ and/or formula E ₂,

Wherein, L ₁ and L' ₁ are each independently a bond, -CHR ₁-、-CO-、-COO-、-S(＝O)₂ -;

Each R ₁ and R' ₁ is independently selected from the group consisting of substituted or unsubstituted: H. amino, C1-C15 alkyl, C1-C15 alkylamino, C6-C15 aryl, C2-C15 alkenyl, C2-C15 alkynyl, C1-C15 alkylhydroxy, C1-C15 alkylaldehyde, C1-C15 alkyl ester, thio C1-C15 alkyl ester, -Rc-COO-Rc ', -Rc-CO-Rc', -Rc-O-Rc '-, -Rc-S-Rc', 5-15 membered heteroaryl, 5-12 membered heterocyclyl;

R _a and R _b are each independently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl, C1-C15 alkylamino, C2-C15 alkenyl, C2-C15 alkynyl, C1-C15 alkylhydroxy, C1-C15 alkylaldehyde, C ₁-C₁₅ alkylsulfonyl, -Rc-COO-Rc ', -Rc-CO-Rc', -Rc-O-Rc '-, -Rc-S-Rc', C3-C12 cycloalkyl, C4-C12 cycloalkenyl, 5-12 membered heterocyclyl, C6-C12 aryl, 5-12 membered heteroaryl;

Or in formula E1, ra together with O and its adjacent C atom forms a substituted or unsubstituted 3-12 membered heterocyclic group containing at least 1O heteroatom and 0-2 heteroatoms selected from N and S;

Or in formula E2, R _a and R _b together with the N atom adjacent thereto form a substituted or unsubstituted 3-12 membered heterocyclyl; or Ra together with N and its adjacent C atom forms a substituted or unsubstituted 3-12 membered heterocyclic group containing at least 1N heteroatom and 0-2 heteroatoms selected from O and S;

Rc is independently selected from the group consisting of substituted or unsubstituted: C1-C15 alkylene, C2-C15 alkenylene, C2-C15 alkynylene, C3-C12 cycloalkylene, C4-C12 cycloalkenylene, 3-12 heterocyclylene, C6-C12 arylene, 5-12 heteroarylene;

rc "is independently selected from the group consisting of substituted or unsubstituted: C1-C15 alkyl, C1-C15 alkylamino, C2-C15 alkenyl, C2-C15 alkynyl, C3-C12 cycloalkyl, C4-C12 cycloalkenyl, 3-C12 heterocyclyl, C6-C12 aryl, 5-C12 heteroaryl;

m is 1,2, 3, or 4;

m' is 1,2, 3, or 4;

q is an integer of 0, 1,2, 3, 4, 5;

q' is an integer of 0, 1, 2, 3; and m '+q'. Ltoreq.4;

Wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, -SH, C1-C6 alkyl S-, ph-, -PhOH, glucosyl, C1-C15 alkyl, C1-C15 haloalkyl, C1-C15 alkoxy, C1-C15 haloalkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl, 5-10 membered heteroaryl, 5-12 membered heterocyclyl.

In another preferred embodiment, rc "is methyl.

In another preferred embodiment, R ₁ and R' ₁ are each independently a substituted or unsubstituted C1-C3 alkylamino group, preferably methylamino.

In another preferred embodiment, the polymer is a copolymer.

In another preferred embodiment, the polymer is a homopolymer.

In another preferred embodiment, the polymer is polymerized with an initiator selected from the group consisting of: methyl triflate, trifluoromethyl benzoyl, substituted or unsubstituted C1-C15 haloalkyl.

In another preferred example, when the polymer is a homopolymer of a repeating unit represented by formula E2, each R ₁ 'is H, L ₁' is CH ₂ and m '+q' =4, ra and Rb are not both H.

In another preferred embodiment, L ₁ and L' ₁ are each independently-CH ₂ -.

In another preferred embodiment, the amino or guanidino functional group in the oxazoline polymer structure is in the form of a hydrochloride, hydrobromide, formate, acetate, or trifluoroacetate salt.

In another preferred embodiment, the amino groups in the oxazoline polymer structure are converted to guanidino functionality.

In another preferred embodiment, the oxazoline polymer structure is a ternary, quaternary and higher random or block copolymer.

In another preferred embodiment, the ends of the oxazoline polymer structure are converted to other functional groups such as hydroxyl, amino, ester groups, and the like.

In another preferred example, after the end of the oxazoline polymer structure is converted into other functional groups such as hydroxyl, amino, ester, etc., the end of the oxazoline polymer structure is modified by active molecules such as other dye molecules, drug molecules, etc., to obtain the oxazoline polymer with the end functionalized.

In another preferred embodiment, R _a and R _b are each independently H, benzyl, t-butoxycarbonyl, benzyloxycarbonyl, t-butyl, trityl, fluorenylmethoxycarbonyl; or R _a and R _b together with the N atom adjacent thereto form a succinimide-containing group.

In another preferred embodiment, the succinimide-containing group is selected from the group consisting of: wherein/> Represents a single bond or a double bond, ar is a substituted or substituted 5-12 membered heteroaryl group or a substituted or substituted C6-C12 aryl group, wherein the substitution refers to substitution with one or more groups selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, 5-10 membered heteroaryl, 5-12 membered heterocyclyl.

In another preferred embodiment, formula E ₂ has the structure shown in formula E' ₂:

Wherein,

Ring a is independently a substituted or unsubstituted 3-12 membered heterocyclyl;

wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, -SH, C1-C6 alkyl S-, ph-, -PhOH, glucosyl, C1-C15 alkyl, C1-C15 haloalkyl, C1-C15 alkoxy, C1-C15 haloalkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl, 5-10 membered heteroaryl, 5-12 membered heterocyclyl;

r '₁, m', rb are as defined above.

In another preferred embodiment, the oxazoline polymer is a homopolymer comprising a repeat unit of either formula E ₁ or formula E ₂.

In another preferred embodiment, the oxazoline polymer is a copolymer comprising a plurality of repeating units represented by formula E ₁ or formula E ₂.

In another preferred embodiment, the oxazoline polymer has an average molecular weight in the range of 500 to 100000, preferably 1000 to 10000, more preferably 2000 to 4000.

In another preferred embodiment, the oxazoline polymer has a PDI in the range of 1.01 to 1.5.

In another preferred embodiment, the oxazoline polymer is modified at both ends with functional groups or reactive molecules, respectively.

In another preferred embodiment, the side chains of the oxazoline polymer are modified with functional groups or reactive molecules.

In another preferred embodiment, the side chains of the oxazoline polymer are used as an initiator to initiate polymerization of other monomers to obtain the oxazoline polymer brush.

In another preferred embodiment, the oxazoline polymer has a pendant amino group as an initiator, and the other monomer to be initiated is selected from the group consisting of: alpha-NCA, alpha-NTA, beta-NCA, beta-NTA, gamma-NCA, gamma-NTA; α -NNCA, α -NNTA, β -NNCA, β -NNTA, γ -NNCA, γ -NNTA (NCA as described herein is an N-carboxyanhydride monomer, NNCA is an N-substituted N-carboxyanhydride monomer, NTA is an N-carboxythiocarbonylanhydride monomer, NNTA is an N-substituted N-carboxythiocarbonylanhydride monomer, wherein α -, β -, γ -refer to α -amino acids, β -amino acids, γ -amino acids, respectively).

In another preferred embodiment, the other monomer is one or more of the compounds of formula D, E, F:

X ₂ = O or S

Wherein X ₂ is O or S; r ₁₁ and R ₂₂ may be independently selected from optionally substituted functional groups.

In another preferred embodiment, the oxazoline polymer further comprises one or more of the repeating units B and/or C:

Wherein,

R ₂ is independently selected from the group consisting of substituted or unsubstituted: C1-C8 alkyl, C3-C8 cycloalkyl, C6-C12 aryl, C6-C12 arylalkyl, C2-C15 alkenyl, C4-C12 cycloalkenyl, C2-C15 alkynyl, C1-C15 alkylhydroxy, C1-C15 alkylaldehyde, -Rc-COO-Rc ', -Rc-CO-Rc', -Rc-O-Rc '-, -Rc-S-Rc', 5-12 membered heteroaryl, 5-12 membered heterocyclyl;

R _a is independently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl, C6-C12 aryl, C2-C15 alkenyl, C2-C15 alkynyl, C3-C12 cycloalkyl, C4-C12 cycloalkenyl, 5-12 membered heteroaryl, 5-12 membered heterocyclyl-CH ₂ -;

L ₁、R₁, m and q are as defined above.

In another preferred embodiment, when the oxazoline polymer is a binary copolymer having repeating units of formula E ₂ and formula B, R ₂ is not H, methyl or ethyl.

In another preferred embodiment, the oxazoline polymer does not contain the following repeating units:

Wherein t is a positive integer not less than 1.

In another preferred embodiment, the repeat unit of formula E ₁, formula E ₂, formula B, or formula C may optionally be linked to another repeat unit of formula E ₁, formula E ₂, formula B, or formula C.

In another preferred example, the oxazoline polymer is an oxazoline polymer having an amino group in a side chain, and has a structure as shown in formula II:

Wherein n is an integer of 5 to 50000; 0% < x <100%; y is more than or equal to 0% and less than 100%; z is more than or equal to 0 percent and less than 100 percent, wherein, the calculation modes of x, y and z are the corresponding number of side chain repeated units divided by the total number of repeated units;

m, m', R ₁、R'₁、R₂、R_a and R _b are as defined above.

In another preferred embodiment, 10% or less of x+y+z or less than 100%, more preferably 50% or less of x+y+z or less than 100%.

In another preferred embodiment, 30% or more and 90% or less of x.

In another preferred embodiment, 10% or less y <70%.

In another preferred embodiment, 10% or less of z <70%.

In another preferred embodiment n is 5 to 5000, preferably 20 to 1000.

In another preferred example, the oxazoline polymer having an amino group in a side chain has a structure as shown in formula V:

Wherein n, x, y, z, m, m', R ₁、R'₁ and R ₂ are as defined above.

In another preferred embodiment, the oxazoline polymer having an amino group in a side chain has a structure as shown in formula VII:

n, x, y, z, A, m, m', R ₁、R'₁、R₂ and R _b are as defined above.

In another preferred embodiment, the oxazoline polymer having an amino group in a side chain thereof has a structure as shown in formula VIII:

R ₆ is independently substituted or unsubstituted C1-C15 alkyl-NRaRb, substituted or unsubstituted 5-12 membered heterocyclyl;

wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, -SH, C1-C6 alkyl S-, ph-, -PhOH, 5-12 membered heteroaryl, 5-12 membered heterocyclyl;

n, x, y, z, m, R ₁ and R ₂ are as defined above.

In another preferred example, the oxazoline polymer is an oxazoline polymer having a hydroxyl group in a side chain, and has a structure as shown in formula I:

Wherein, m and R ₁、R₂、R_a are defined as above; n is an integer of 5 to 50000; 0% < x <100%; y is more than or equal to 0% and less than 100%; z is more than or equal to 0% and less than 100%; wherein, the calculation mode of x, y and z is the corresponding number of side chain repeating units divided by the total number of repeating units.

In another preferred embodiment, the oxazoline polymer having hydroxyl groups in the side chains has a structure as shown in IV:

Wherein n, x, y, z, m, R ₁ and R ₂ are defined as described above.

In another preferred example, the oxazoline polymer is an oxazoline polymer having an amino group and a hydroxyl group in a side chain, and has a structure as shown in formula III:

Wherein 0% < w <100%; x ₁ is more than or equal to 0% and less than 100%; wherein, the calculation mode of w, x ₁, y and z is the corresponding repeated unit number divided by the total repeated unit number; ;

m, m', R ₁、R'₁、R₂、R_a、R_b, n, y and z are as defined above.

In another preferred embodiment, 10% w.ltoreq.90%.

In another preferred embodiment, 10% or more and ₁% or less and 90% or less.

In another preferred embodiment, 10% or less w+x1+y+z or less than 100%, more preferably 50% or less w+x1+y+z or less than 100%.

In another preferred embodiment, the oxazoline polymer having an amino group and a hydroxyl group in the side chain has a structure as shown in formula VI or formula IX:

Wherein,

M, m', n, x, y, z, w, x ₁、R₁、R'₁、R₂ and R ₆ are as defined above.

In another preferred embodiment, R ₆ is a moiety other than a carboxyl group in the molecular structure of a natural amino acid or a non-natural amino acid.

In another preferred embodiment, the natural amino acid is selected from the group consisting of: glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartic acid, asparagine, glutamic acid, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine and the like, and derivatives derived from the above amino acids.

In another preferred embodiment, the unnatural amino acid comprises a beta amino acid, a gamma amino acid, a delta amino acid, an epsilon amino acid.

In another preferred embodiment, the unnatural amino acid comprises D-, L-, or DL-form.

In a second aspect of the present invention, there is provided a process for producing an oxazoline polymer having an amino group in a side chain, comprising the steps of:

in an organic solvent, under the existence of an initiator and heating conditions, carrying out polymerization reaction on an oxazoline monomer represented by a formula E ' ₂ and optionally a monomer of a formula B ' and/or a formula C ' so as to obtain an oxazoline polymer with an amino-containing side chain;

Wherein R' ₁、m'、q'、L₁'、R₁、m、q、L₁、R_a and R _b are as defined above.

In another preferred example, the oxazoline polymer with an amino group in a side chain is an oxazoline homopolymer with an amino group in a side chain or an oxazoline copolymer with an amino group in a side chain.

In another preferred example, the oxazoline polymer with the side chain containing the amino group is a block type oxazoline copolymer with the side chain containing the amino group or a blend type oxazoline copolymer with the side chain containing the amino group.

In another preferred embodiment, the method for preparing an oxazoline homopolymer having an amino group in a side chain comprises the steps of: in an organic solvent, in the presence of an initiator, an oxazoline monomer represented by a formula E' ₂ is subjected to polymerization reaction, so that an oxazoline homopolymer with an amino group on a side chain is obtained.

In another preferred example, in the preparation method of the oxazoline polymer with the side chain containing amino groups, the heating condition is in a temperature range of 80-140 ℃; preferably 120 ℃.

In another preferred example, in the preparation method of the oxazoline polymer with the side chain containing amino groups, the polymerization reaction time is 2-8 hours; preferably 6h.

In another preferred embodiment, the method for preparing an oxazoline polymer having amino groups in the side chains, the organic solvent is selected from the group consisting of: DMAc, DMF, or a combination thereof; DMAc is preferred.

In another preferred embodiment, the method for preparing oxazoline polymer having amino groups in the side chains, the initiator is selected from the group consisting of: meOTf, bzOTf, or a combination thereof.

In another preferred embodiment, the oxazoline polymer of the present invention having amino groups in the side chains is prepared by the following method: under the protection of an organic solvent and inert gas, under the condition of the existence of an initiator and heating, the oxazoline monomer represented by a formula E ' ₂ and the optional monomer of a formula B ' and/or a formula C ' are subjected to polymerization reaction, so that the oxazoline polymer with the side chain containing amino is obtained.

In another preferred embodiment, the process for producing the oxazoline polymer having an amino group in a side chain is carried out under the protection of an inert gas.

In another preferred embodiment, the method for preparing the oxazoline polymer with the amino group in the side chain comprises the step of using N ₂ or Ar as inert gas.

In another alternative, the oxazoline monomer of formula E' ₂ has a structure as shown in formula E "₂:

In the method, in the process of the invention,

R ₇ is independently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl, C1-C15 alkylamino, C2-C15 alkenyl, C2-C15 alkynyl, C1-C15 alkylhydroxy, C1-C15 alkylaldehyde, -Rc-COO-Rc ', -Rc-CO-Rc', -Rc-O-Rc '-, -Rc-S-Rc', C3-C12 cycloalkyl, C4-C12 cycloalkenyl, 5-12 membered heterocyclyl, C6-C12 aryl, 5-12 membered heteroaryl;

Rc' is independently selected from the group consisting of substituted or unsubstituted: C1-C15 alkyl, C1-C15 alkylamino, C2-C15 alkenyl, C2-C15 alkynyl, C3-C12 cycloalkyl, C4-C12 cycloalkenyl, 3-C12 heterocyclyl, C6-C12 aryl, 5-C12 heteroaryl;

P ₁ is an amino protecting group selected from the group consisting of: t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), fluorenylmethoxycarbonyl (Fmoc), phthaloyl (Pht), acetyl (Ac), trifluoroacetyl (Tfa), benzyl (Bn), triphenylmethyl (Tr), or R ₇ and P ₁ together with the N atoms adjacent thereto form a succinimide-containing group;

m ' and R ' ₁ are as defined above, and- (CHR ' ₁)_m' -is not- (CH ₂)₄).

In another preferred example, one or more oxazoline monomers represented by the formula E' ₂ are polymerized in an organic solvent in the presence of an initiator to obtain an oxazoline polymer with an amino protecting group in a side chain; removing the protected amino group to obtain an oxazoline polymer with side chain amino groups; optionally, the amino groups in the oxazoline polymer from which the side chain amino groups are derived are functionalized towards guanidine groups to yield the oxazoline polymer from which the side chain guanidine groups are derived.

In another preferred embodiment, the method for preparing the oxazoline copolymer with the side chain containing the amino group comprises the following steps:

(i) In an organic solvent, under the condition of the existence of an initiator and heating, firstly, an oxazoline monomer with a structure of a formula E' ₂ is subjected to polymerization reaction,

(Ii) After the polymerization reaction of the step (i) is finished, adding another monomer with a formula E ' ₂, a formula B ' or a formula C ' to perform polymerization reaction,

And optionally (iii) repeating step (ii) f times, thereby forming a block side chain containing amino oxazoline copolymer;

Or (i ') mixing an oxazoline monomer having a structure of formula E' ₂ with one or more monomers of another formula E '₂, formula B' or formula C 'in an organic solvent, (ii') carrying out polymerization reaction in the presence of an initiator under heating conditions, thereby forming a copolymer with a blend-type side chain containing an amino oxazoline;

Wherein f is an integer not less than 1.

In a third aspect of the present invention, there is provided an oxazoline monomer having a side chain comprising a hydroxyl group, having a structure as shown in formula E' ₁:

Wherein, R ₁、m、q、L₁ and Ra are defined as above.

In another preferred embodiment, formula E' ₁ has the structure shown as formula E "₁:

Wherein P ₂ is a hydroxy protecting group, preferably TMS, TBS, TBDPS, t-Bu; m, R ₁ are as defined above;

In another preferred embodiment, a method for synthesizing an oxazoline monomer shown in E "₁ is provided, which specifically comprises the steps of:

Wherein, m and P ₂、R₁ are defined as above;

X is a leaving group;

(2) Compound 2 was reacted with a base in a second inert solvent to give the monomer of formula E "₁.

In another preferred embodiment, the base is an organic base, an inorganic base, or a combination thereof.

In another preferred embodiment, the base is selected from the group consisting of: sodium hydroxide, potassium hydroxide, triethylamine, N-diisopropylethylamine, or a combination thereof.

In another preferred embodiment, the second inert solvent is independently selected from the group consisting of: methanol, ethanol, acetonitrile, or a combination thereof.

In another preferred embodiment, the leaving group is selected from the group consisting of: br, cl, I, OTs, OMs.

In another preferred embodiment, the method for preparing the monomer of formula E "₁ further comprises the steps of:

In the method, in the process of the invention,

R ₁、P₂, X, m are as defined above;

(1) Compound 1 was reacted with H ₂N(CH₂)₂ X in a first inert solvent to give compound 2 of formula.

In another preferred embodiment, in the step (1), the reaction temperature is 0 to 70 ℃; preferably, it is from 0℃to 25 ℃.

In another preferred embodiment, in the step (1), the reaction time is 6 hours to 1 day.

In another preferred embodiment, in the step (1), H ₂N(CH₂)₂ X is in the form of its hydrochloride, hydrobromide.

In another preferred embodiment, the step (1) is to dissolve the compound 1 and DIEA in a first inert solvent, add Et ₃ N, EDCI and HOBT under ice bath, stir and react for 30min, and then add H ₂N(CH₂)₂ X; after the reaction at room temperature for 8 hours, the reaction mixture is washed twice with deionized water and 5% citric acid aqueous solution respectively, and the organic phase is dried and concentrated, and then is separated and purified by a column to obtain the compound 2.

In another preferred embodiment, in the step (1), the first inert solvent is independently selected from the group consisting of: DCM, DMF, THF, DMAc, or a combination thereof; DCM is preferred.

In another preferred example, the step (2) is to dissolve the compound 2 obtained in the step (1) in a second inert solvent, add a base to react, and under the protection of inert gas at 70 ℃; after the reaction, the solvent is removed, dichloromethane is added for dissolution, deionized water is used for washing for 2 times, and the organic phase is dried and concentrated, distilled and purified under reduced pressure to obtain a monomer of a formula E' ₁.

In another preferred embodiment, in the step (2), the second inert solvent is dried.

In a fourth aspect of the present invention, there is provided a method for preparing an oxazoline polymer having a hydroxyl group in a side chain, comprising the steps of:

Under the protection of inert solvent and inert gas, under the condition of the existence of an initiator and heating, carrying out polymerization reaction on an oxazoline monomer represented by a formula E ' ₁ and optionally a monomer of a formula B ' and/or a formula C ', thereby obtaining an oxazoline polymer with a side chain hydroxyl group;

Wherein R ₁、m、q、L₁、R_a and R ₂ are as defined above.

In another preferred example, the oxazoline polymer with a hydroxyl group in a side chain is an oxazoline homopolymer with a hydroxyl group in a side chain or an oxazoline copolymer with a hydroxyl group in a side chain.

In another preferred example, the oxazoline polymer with the side chain containing the hydroxyl group is a block type oxazoline copolymer with the side chain containing the hydroxyl group or a blend type oxazoline copolymer with the side chain containing the hydroxyl group.

In another preferred embodiment, the method for preparing an oxazoline homopolymer having a hydroxyl group in a side chain comprises the steps of: under the protection of inert solvent and inert gas, under the condition of the existence of an initiator and heating, an oxazoline monomer represented by a formula E' ₁ is subjected to polymerization reaction, so that an oxazoline homopolymer with a side chain containing hydroxyl is obtained.

In another preferred embodiment, in the process for the preparation of oxazoline polymers having hydroxyl groups in the side chains, the inert solvent is selected from the group consisting of: DMAc, DMF, meCN, phCN, or a combination thereof.

In another preferred example, in the method for producing an oxazoline polymer having hydroxyl groups in the side chains, the inert gas is selected from the group consisting of: n ₂, ar, or a combination thereof.

In another preferred embodiment, in the process for the preparation of oxazoline polymers having hydroxyl groups in the side chains, the initiator is selected from the group consisting of: meOTf, bzOTf, CH ₃(CH₂)₅Br、CH₃(CH₂)₅ OTs, or a combination thereof.

In another preferred embodiment, the method for preparing the oxazoline copolymer with the side chain containing hydroxyl comprises the following steps:

(a) Under the protection of inert solvent and inert gas, under the condition of initiator existence and heating, first, an oxazoline monomer with a structure of formula E' ₁ is polymerized,

(B) After the polymerization reaction of the step (a) is finished, adding another monomer with a formula E ' ₁, a formula B ' or a formula C ' to perform polymerization reaction,

And optionally (c) repeating step (b) f times, thereby forming a block type oxazoline copolymer having hydroxyl groups in the side chains;

Or (a ') mixing an oxazoline monomer with a structure shown in a formula E' ₁ with one or more monomers shown in a formula E '₁, a formula B' or a formula C 'in the presence of an initiator and under the protection of inert gases, and (B') carrying out polymerization reaction under the condition of heating so as to form a copolymer with a blend-type side chain containing the amino oxazoline;

f is an integer greater than or equal to 1.

In another preferred example, in the preparation method of the oxazoline polymer with hydroxyl group in the side chain, the heating condition is in the temperature range of 80-140 ℃; preferably from 80℃to 120 ℃.

In another preferred example, in the method for producing an oxazoline polymer having a hydroxyl group in a side chain, the polymerization reaction is carried out for a period of 2 hours to 8 hours; preferably 6h.

In a fifth aspect of the present invention, there is provided a method for preparing an oxazoline polymer having an amino group and a hydroxyl group in a side chain, comprising the steps of:

In an organic solvent, under the existence of an initiator and heating conditions, carrying out polymerization reaction on an oxazoline monomer represented by a formula E '₂, an oxazoline monomer represented by a formula E' ₁ and optionally oxazoline monomers with other structures, so as to obtain an oxazoline polymer with amino groups and hydroxyl groups on side chains;

In another preferred embodiment, optionally the oxazoline monomer of other structure is a monomer of formula B 'or formula C',

Wherein R ₁、R₂、m、q、L₁, and R _a are defined as described above.

In another preferred example, the oxazoline polymer with the side chain containing the amino group and the hydroxyl group is a block type oxazoline copolymer with the side chain containing the amino group or a blend type oxazoline copolymer with the side chain containing the amino group.

In another preferred example, in the preparation method of the oxazoline polymer with the side chain containing amino groups and hydroxyl groups, the heating condition is in a temperature range of 80-140 ℃; preferably 120 ℃.

In another preferred example, in the preparation method of the oxazoline polymer with the side chain containing amino groups and hydroxyl groups, the polymerization reaction time is 2-36 h; preferably 6h.

In another preferred embodiment, the method for preparing oxazoline polymer having amino and hydroxyl groups in the side chains, the organic solvent is selected from the group consisting of: DMAc, DMF, or a combination thereof; DMAc is preferred.

In another preferred embodiment, the method for preparing oxazoline polymer having amino and hydroxyl groups in the side chains, wherein the initiator is selected from the group consisting of: meOTf, bzOTf, or a combination thereof.

In another preferred embodiment, the process for producing the oxazoline polymer having an amino group and a hydroxyl group in its side chain is carried out under the protection of an inert gas.

In another preferred embodiment, the method for preparing oxazoline polymer with amino and hydroxyl groups on the side chains comprises the step of preparing the oxazoline polymer with the side chains, wherein the inert gas is N ₂, ar or a combination thereof.

In another preferred example, one or more oxazoline monomers represented by the formula E "₂ and one or more oxazoline monomers represented by the formula E" ₁ are polymerized in an organic solvent in the presence of an initiator and under heating, thereby obtaining an oxazoline polymer with a side chain containing amino groups protected and hydroxyl groups protected; removing the protecting group to obtain oxazoline polymer with side chain amino and hydroxyl; optionally, the amino groups in the oxazoline polymer from which the amino groups and the hydroxyl groups are obtained are functionalized towards the guanidine groups to obtain the oxazoline polymer from which the side chain guanidine groups and the hydroxyl groups are obtained.

In another preferred example, the oxazoline polymer with the side chain containing the amino group and the hydroxyl group is a block type oxazoline copolymer with the side chain containing the amino group and the hydroxyl group or a blend type oxazoline copolymer with the side chain containing the amino group and the hydroxyl group.

In another preferred embodiment, the preparation method of the oxazoline copolymer with the side chain containing amino and hydroxyl comprises the following steps:

(d) In an organic solvent, under the condition of the existence of an initiator and heating, firstly, an oxazoline monomer with a structure of a formula E' ₂ is subjected to polymerization reaction,

(Dd) after the polymerization reaction in step (d) is completed, adding another monomer with the formula E' ₁ to perform polymerization reaction,

(Ddd) after the polymerization reaction in the step (dd) is completed, adding another monomer with the formula B 'or the formula C' to perform the polymerization reaction,

And optionally (dddd) repeating steps (d), (dd) or (ddd) f times, thereby forming a block-type copolymer having amino and hydroxyl oxazoline groups in the side chains;

Or (d ') mixing oxazoline monomers having the structures of formula E' ₂ and formula E '₁ with one or more monomers of another formula E' ₂, formula E '₁, formula B' or formula C 'in an organic solvent, (ii') polymerizing in the presence of an initiator and under heating to form a copolymer having amino groups and hydroxyl oxazolines in the side chains of the copolymer;

Wherein f is an integer not less than 1.

In a sixth aspect of the invention there is provided the use of an oxazoline polymer having amino groups in the side chains for the preparation of materials for antibacterial, antitumor, cell adhesion, tissue engineering, drug and gene delivery, drug modification, cell adhesion or self-assembly.

In another preferred embodiment, the antimicrobial material is in the form of a solution, a surface coating.

In another preferred example, the antibacterial object is a microorganism such as bacteria and fungi, and may include escherichia coli (e.coli), pseudomonas (p.aeromonas), acinetobacter baumannii (a.baumannii), enterobacter aerogenes (e.aerogenes), klebsiella pneumoniae (k.pneumoniae), serratia marcescens (s.marcescens), enterobacter cloacae (e.cloacae), bacillus subtilis (b.sublis), staphylococcus aureus (s.aureus), staphylococcus epidermidis (s.epidis), candida albicans (c.albicans), and cryptococcus neoformans (c.neoformans).

In another preferred embodiment, the antimicrobial uses include the release of cells, biofilms, spores and the like from microorganisms.

In another preferred embodiment, the polymer is used for drug synergistic antibacterial.

In another preferred embodiment, the polymer is used for treating tumors.

In another preferred embodiment, the tumor is selected from the group consisting of: melanoma, skin cancer, glioma, mesothelioma, lymphoma, leukemia, breast cancer, ovarian cancer, cervical cancer, glioblastoma, multiple myeloma, prostate cancer, burkitt's lymphoma, head and neck cancer, colon cancer, colorectal cancer, non-small cell lung cancer, esophageal cancer, gastric cancer, pancreatic cancer, hepatobiliary cancer, gall bladder cancer, small intestine cancer, rectal cancer, kidney cancer, bladder cancer, prostate cancer, penile cancer, urinary tract cancer, testicular cancer, vaginal cancer, uterine cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, pancreatic endocrine cancer, carcinoid, bone cancer, retinoblastoma, hodgkin's lymphoma, non-hodgkin's lymphoma, multicenter kaposi's disease, AIDS-related primary exudative lymphoma, neuroectodermal tumor or rhabdomyosarcoma.

The seventh aspect of the invention provides an application of an oxazoline polymer with a side chain containing hydroxyl, which is used in the biomedical fields of surface antifouling, cell freezing, drug modification and the like.

In another preferred embodiment, the oxazoline polymer of pendant hydroxyl groups is used in a surface anti-fouling coating.

In another preferred example, the surface anti-fouling object is adhesion of bacteria including escherichia coli (e.coli), pseudomonas (p. Aeromonas), acinetobacter baumannii (a.baumannii), enterobacter aerogenes (e.aerogenes), klebsiella pneumoniae (k.pneumannia), serratia marcescens (s.marcescens), enterobacter cloacae (e.cloacae), bacillus subtilis (b.subtis), staphylococcus aureus (s.aureus), staphylococcus epidermidis (s.epididitis), candida albicans (c.albicans), and cryptococcus neoformans (c.neoformans).

In another preferred embodiment, the anti-fouling use comprises adhesion to proteins.

In another preferred embodiment, the oxazoline polymer of pendant hydroxyl groups is used for cell cryopreservation.

In another preferred embodiment, the cell is selected from the group consisting of: mammalian embryonic stem cells, mammalian induced pluripotent stem cells, mammalian primary pluripotent stem cells, cells differentiated from mammalian embryonic stem cells, mammalian induced pluripotent stem cells and mammalian primary pluripotent stem cells, mammalian cells reprogrammed from other cell types, mammalian primary cells, human umbilical vein endothelial cells, cancer cells, T cells, mammalian tissue stem cells, mammalian cell lines.

According to an eighth aspect of the invention, the application of the oxazoline polymer with the side chain containing amino and hydroxyl is provided, and the oxazoline polymer can be used in the biomedical fields of antibiosis, anti-tumor, tissue engineering, medicine and gene delivery, medicine modification, cell adhesion, self-assembly materials, surface antifouling, cell freezing storage, medicine modification and the like.

In a ninth aspect of the invention there is provided a polymerization process comprising using the oxazoline polymer of the first aspect as an initiator to initiate a monomer.

In another preferred embodiment, in the polymerization method, the oxazoline polymer is a polymer having an amino group in a side chain.

In another preferred embodiment, in the polymerization method, the oxazoline polymer is a polymer having-NH ₂ in a side chain.

In another preferred embodiment, in the polymerization method, the oxazoline polymer has a structure as shown in formula V:

Wherein n, x, y, z, m, m', R ₁、R'₁ and R ₂ are as defined above.

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a structure as shown in formula VII:

n, x, y, z, A, m, m', R ₁、R'₁、R₂ and R _b are as defined above.

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a structure as shown in formula VIII:

n, x, y, z, m, R ₁ and R ₂ are as defined above.

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a structure as shown in formula I:

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a structure as shown in IV:

Wherein n, x, y, z, m, R ₁ and R ₂ are defined as described above.

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a structure as shown in formula VI or formula IX:

Wherein,

In another preferred embodiment, in the polymerization process, the oxazoline polymer has a pendant amino group as an initiator, and the other monomer is initiated from the group consisting of: alpha-NCA, alpha-NTA, beta-NCA, beta-NTA, gamma-NCA, gamma-NTA; α -NNCA, α -NNTA, β -NNCA, β -NNTA, γ -NNCA, γ -NNTA (NCA as described herein is an N-carboxyanhydride monomer, NNCA is an N-substituted N-carboxyanhydride monomer, NTA is an N-carboxythiocarbonylanhydride monomer, NNTA is an N-substituted N-carboxythiocarbonylanhydride monomer, wherein α -, β -, γ -refer to α -amino acids, β -amino acids, γ -amino acids, respectively).

X ₂ = O or S

It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute new or preferred technical solutions. And are limited to a space, and are not described in detail herein.

Detailed Description

The present inventors have conducted long and intensive studies and have unexpectedly found that oxazoline polymers comprising repeating units represented by the formula E ₁ and/or the formula E ₂ can be widely used in biomedical fields such as antibacterial, drug delivery, drug modification, cell adhesion, gene delivery, self-assembled materials, antitumor, cell adhesion, tissue engineering, surface antifouling, cell cryopreservation, and the like. Based on the above findings, the inventors have completed the present invention.

Specifically, the inventor prepares the oxazoline polymer with amino groups and/or hydroxyl groups on the side chains by optimizing polymerization conditions (under the protection of inert gas, methyl triflate or benzoyl triflate is used as an initiator, N-2 methylacetamide is used as a preferable solvent, and under the heating condition).

The oxazoline polymer with the side chain containing amino can be used in the fields of antibacterial, antifungal, antiviral, anti-mite, anti-tumor, cell adhesion, tissue engineering, drug modification, protein protection, drug synergy, drug delivery, gene delivery, self-assembly materials and the like.

The invention also synthesizes an oxazoline monomer with a side chain containing protected hydroxyl, and the oxazoline polymer with the side chain hydroxyl prepared by polymerization reaction can be used in the fields of surface antifouling, protein modification and protection, cell protection, tissue and organ cryoprotection, drug modification and the like.

The oxazoline polymer with the side chain containing the amino group and the hydroxyl group has the functions of the oxazoline polymer with the side chain containing the amino group and the hydroxyl group.

Terminology

In the present invention, unless otherwise indicated, terms used have the ordinary meanings known to those skilled in the art.

When substituents are described by conventional formulas written from left to right, the substituents also include chemically equivalent substituents obtained when writing formulas from right to left. For example, -CH ₂ O-is equivalent to-OCH ₂ -.

Throughout the specification, the terms "optionally substituted" or "may be substituted" and the like mean that the group may or may not be further substituted or fused with one or more non-hydrogen substituents (to form a polycyclic ring system). Substituents for suitable chemically suitable specific functional groups will be apparent to those skilled in the art.

As used herein, the term "alkyl" refers to a straight or branched chain alkyl group containing several carbon atoms, wherein "C ₁-C₁₅ alkyl" refers to a straight or branched chain alkyl group having 1 to 15 carbon atoms, including alkyl groups of 1,2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 carbon atoms, alkyl groups preferably being, for example, C₁-C₂、C₁-C₃、C₁-C₄、C₁-C₅、C₁-C₆、C₁-C₇、C₁-C₈、C₁-C₉、C₁-C₁₀、C₂-C₃、C₂-C₄、C₂-C₅、C₂-C₆、C₃-C₄、C₃-C₅、C₃-C₆、C₃-C₇、C₃-C₈、C₄-C₅、C₄-C₆ or C _5-6. Typical "alkyl" groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, and,Pentyl, isopentyl, heptyl, 4-dimethylpentyl, octyl, 2, 4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl and the like. In the present invention, alkyl groups also include substituted alkyl groups. "substituted alkyl" means that one or more positions in the alkyl group are substituted, especially 1 to 4 substituents, and may be substituted at any position.

As used herein, the term "C ₁-C₁₅ alkoxy" refers to a straight or branched chain alkoxy group having 1 to 15 carbon atoms, having a C ₁-C₁₅ alkyl-O-structure, and C ₁-C₁₅ alkoxy includes, but is not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, and the like, preferably ethoxy. C ₁-C₁₅ alkoxy also includes substituted C ₁-C₁₅ alkoxy.

As used herein, the term "C ₁-C₁₅ alkylhydroxy" refers to-C ₁-C₁₅ alkylene-OH, -C ₁-C₁₅ alkylene having the definition as described above, and C ₁-C₁₅ alkylhydroxy includes, but is not limited to-CH ₂OH、-CH₂CH₂OH.C₁-C₁₅ alkylhydroxy and also includes substituted C ₁-C₁₅ alkylhydroxy.

As used herein, the term "C ₁-C₁₅ alkylsulfonyl" refers to C ₁-C₁₅ alkyl S (=o) ₂ -.

As used herein, the term "C ₁-C₁₅ alkyl-C ₆-C₁₅ aryl" refers to a-C ₁-C₁₅ alkyl-C ₆-C₁₅ aryl, such as-CH ₂CH₂CH₂ Ph, -Bn.

As used herein, the term "C ₁-C₁₅ alkyl ester" refers to a C ₁-C₁₅ alkyl C (=o) -O-or- (=o) -O-C ₁-C₁₅ alkyl.

As used herein, the term "thio C1-C15 alkyl ester group" refers to a C ₁-C₁₅ alkyl C (=s) -O-or- (=s) -O-C ₁-C₁₅ alkyl.

As used herein, the term "guanidino" refers to NHC (=nh) NH-.

As used herein, the term "C ₁-C₁₅ alkylcarboxy" refers to-C ₁-C₁₅ alkylcarboxyl, for example -CH₂COOH、-CH₂CH₂COOH、-CH₂CH₂CH₂COOH、-CH₂CH₂CH₂CH₂COOH, preferably-CH ₂COOH、-CH₂CH₂ COOH. Wherein the alkyl group may be substituted.

As used herein, the term "C ₁-C₁₅ alkyl aldehyde" refers to-C ₁-C₁₅ alkyl CHO, for example -CH₂CHO、-CH₂CH₂CHO、-CH₂CH₂CH₂CHO、-CH₂CH₂CH₂CH₂CHO, preferably-CH ₂CHO、-CH₂CH₂ CHO. Wherein the alkyl group may be substituted.

As used herein, the term "C ₁-C₁₅ alkylamino" refers to-C ₁-C₁₅ alkyl-NH ₂, e.g., -CH₂-NH₂、-CH₂CH₂-NH₂、-CH₂CH₂CH₂-NH₂、-CH₂CH₂CH₂CH₂-NH₂, is preferably-CH ₂-NH₂、-CH₂CH₂-NH₂. Wherein H in the amino group (-NH ₂) may also be substituted.

As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon having at least 2 carbon atoms and at least one double bond. Alkenyl groups may include any number of carbon atoms, where "C2-C15 alkenyl" refers to straight or branched chain hydrocarbons having 2-15 carbon atoms and at least one double bond, such as C₂、C₂-C₃、C₂-C₄、C₂-C₅、C₂-C₆、C₂-C₇、C₂-C₈、C₂-C₉、C₂-C₁₀、C₃、C₃-C₄、C₃-C₅、C₃-C₆、C₄、C₄-C₅、C_4-C₆、C₅、C_5-C₆ and C ₆. Alkenyl groups may have any suitable number of double bonds including, but not limited to, 1,2,3,4, 5 or more. Examples of alkenyl groups include, but are not limited to, vinyl (vinyl group)), propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1, 3-pentadienyl, 1, 4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1, 3-hexadienyl, 1, 4-hexadienyl, 1, 5-hexadienyl, 2, 4-hexadienyl, or 1,3, 5-hexatrienyl. As with the alkyl groups described above, alkenyl groups may be substituted or unsubstituted.

As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon having at least 2 carbon atoms and at least one triple bond. Alkynyl groups may include any number of carbon atoms, "C2-C15 alkynyl" refers to straight or branched chain hydrocarbons having 2 to 15 carbon atoms and at least one triple bond, such as C₂、C₂-C₃、C₂-C₄、C₂-C₅、C₂-C₆、C₂-C₇、C₂-C₈、C₂-C₉、C₂-C₁₀、C₃、C₃-C₄、C₃-C₅、C₃-C₆、C₄、C₄-C₅、C₄-C₆、C₅、C₅-C₆ and C ₆. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1, 3-pentadiynyl, 1, 4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1, 3-hexadiynyl, 1, 4-hexadiynyl, 1, 5-hexadiynyl, 2, 4-hexadiynyl, or 1,3, 5-hexadiynyl. As with the alkyl groups described above, alkynyl groups may be substituted or unsubstituted.

The term "aryl" refers to aromatic cyclic hydrocarbon compounds wherein "C6-C15 aryl" refers to aromatic cyclic hydrocarbon compounds containing 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 ring carbon atoms, having 1 to 5 rings, especially mono-and bi-cyclic groups such as phenyl, biphenyl or naphthyl. The aromatic ring of the aryl group may be linked by a single bond (e.g., biphenyl), or condensed (e.g., naphthalene, anthracene, etc.), where the aromatic ring contains two or more aromatic rings (bicyclic, etc.). "substituted aryl" means that one or more positions in the aryl group are substituted, especially 1 to 3 substituents, and can be substituted at any position.

As used herein, the term "heteroaryl" refers to a heteroaromatic system containing 1 to 3 atoms selected from N, O, S, wherein "5-15 membered heteroaryl" refers to a 5-15 membered heteroaromatic system containing 1 to 3 atoms selected from N, O, S. Heteroaryl is preferably a 5 to 10 membered ring, more preferably 5 or 6 membered ring, and heteroaryl includes, but is not limited to, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazolyl, tetrazolyl, and the like. "heteroaryl" may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, deuteroalkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxy, mercapto, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylthio, oxo, carboxyl, and carboxylate.

As used herein, the term "cycloalkyl" refers to a fully saturated cyclic hydrocarbon group having several carbon atoms, wherein "C ₃-C₁₅ cycloalkyl" refers to a fully saturated cyclic hydrocarbon group having 3 to 15 carbon atoms, preferably C₃-C₄、C₃-C₅、C₃-C₆、C₃-C₇、C₃-C₈、C₃-C₉、C₃-C₁₀." substituted C ₃-C₁₅ cycloalkyl "refers to one or more positions in the cycloalkyl group being substituted, especially 1 to 4 substituents, which may be substituted at any position, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. In the present invention, "cycloalkyl" is intended to include "substituted cycloalkyl".

As used herein, the term "cycloalkenyl" refers to an unsaturated cyclic hydrocarbon group having 1-3 double bonds with several carbon atoms, wherein "C ₄-C₁₅ cycloalkenyl" refers to an unsaturated cyclic hydrocarbon group having 1-3 double bonds with 4-15 carbon atoms, preferably C ₆-C₁₀ cycloalkenyl, C ₄-C₆ cycloalkenyl, cycloalkenyl including but not limited to cyclobutenyl, cyclopentenyl, cyclohexenyl.

As used herein, the term "heterocyclyl" refers to a fully saturated or partially unsaturated cyclic group having several (3 or more) ring atoms and having 1-3 heteroatoms, wherein "5-15 membered heterocyclyl" refers to a fully saturated or partially unsaturated cyclic group having 5-15 ring atoms and having 1-3 heteroatoms (including but not limited to, e.g., 3-7 membered monocyclic, 6-11 membered bicyclic, or 8-12 membered tricyclic ring systems). Wherein the nitrogen or sulfur atom may be oxidized and the nitrogen atom may be quaternized. The heterocyclic group may be attached to any heteroatom or carbon atom residue of a ring or ring system molecule. Typical monocyclic heterocycles include, but are not limited to, azetidinyl, pyrrolidinyl, oxetanyl, pyrazolinyl, imidazolinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, hexahydroazepinyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1, 3-dioxanyl, and tetrahydro-1, 1-dioxythiophene, and the like. Polycyclic heterocyclyl groups include spiro, fused and bridged heterocyclic groups; wherein the heterocyclic groups of the spiro ring, the condensed ring and the bridged ring are optionally connected with other groups through single bonds, or are further connected with other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups through any two or more atoms on the ring in a parallel ring manner; the heterocyclic group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, deuteroalkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxy, mercapto, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylthio, oxo, carboxyl, and carboxylate. Heterocyclyl groups include, but are not limited to: tetrahydropyrrolyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and the like.

When a substituent is a non-terminal substituent or a related group, it is a subunit of the corresponding group, typically a divalent group, e.g., an alkyl group, after removal of one H atom is an alkylene group (e.g., methylene, ethylene, propylene, isopropylene (e.g.)) Butylene (e.g./>) Pentylene (e.g./>) Hexyl (e.g./> ) Heptylene (e.g./>) Etc.), cycloalkyl corresponds to cycloalkylene (e.g.: /(I)Etc.), the heterocyclyl corresponds to the heterocyclylene group (e.g., as: /(I)) Cycloalkyl corresponds to a heterocyclyl group (e.g.: /(I) Etc.), the alkoxy group corresponds to the alkyleneoxy group (-CH₂O-、-CH₂CH₂-O-CH₂-、-CH₂OCH₂CH₂CH₂-), etc.

The term "plurality" as used herein refers to two or more.

As described herein, the compounds of the present invention may be substituted with any number of substituents or functional groups to extend their inclusion. In general, the term "substituted", whether appearing before or after the term "optional", in the context of the present invention includes the general formula of substituents, means that the specified structural substituent is used in place of the hydrogen radical. When multiple of a particular structure are substituted at a position with multiple particular substituents, the substituents may be the same or different at each position. The term "substitution" as used herein includes all permissible organic compound substitutions. In a broad sense, permissible substituents include acyclic, cyclic, branched, unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. In the present invention, the heteroatom nitrogen may have a hydrogen substituent or any of the permissible organic compounds described hereinabove to supplement the valence state thereof. Furthermore, the present invention is not intended to be limited in any way to allow substitution of organic compounds.

In the present invention, unless otherwise specified, the groups include the corresponding substituents and subunits, such as: alkyl includes substituted alkyl, cycloalkyl includes substituted cycloalkyl, aryl includes substituted aryl, heteroaryl includes substituted heteroaryl, heterocyclyl includes substituted heterocyclyl, and the like. Those skilled in the art will appreciate that combinations of substituents contemplated by the present invention are those that are stable or chemically achievable. Typical substitutions include, but are not limited to, one or more of the following groups: such as hydrogen, deuterium, halogen (e.g., a single halogen substituent or multiple halogen substituents, the latter such as trifluoromethyl or an alkyl group comprising Cl ₃), nitrile, nitro, oxygen (e.g., =o), trifluoromethyl, trifluoromethoxy, cycloalkyl, C2-C6 alkenyl, C4-C10 cycloalkenyl, C2-C6 alkynyl, heterocyclyl, aryl, heteroaryl 、OR_a、SR_a、S(＝O)R_e、S(＝O)₂R_e、P(＝O)₂R_e、S(＝O)₂OR_e,P(＝O)₂OR_e、NR_bR_c、NR_bS(＝O)₂R_e、NR_bP(＝O)₂R_e、S(＝O)₂NR_bR_c、P(＝O)₂NR_bR_c、C(＝O)OR_d、C(＝O)R_a、C(＝O)NR_bR_c、OC(＝O)R_a、OC(＝O)NR_bR_c、NR_bC(＝O)OR_e,NR_dC(＝O)NR_bR_c、NR_dS(＝O)₂NR_bR_c、NR_dP(＝O)₂NR_bR_c、NR_bC(＝O)R_a、, or NR _bP(＝O)₂R_e, wherein R _a as present herein may independently represent hydrogen, deuterium, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C3-C10 cycloalkenyl, C2-C6 alkynyl, 3-8 heterocyclyl, 5-14 membered heteroaryl, or C6-C14 aryl, R _b、R_c and R _d may independently represent hydrogen, deuterium, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, 5-14 membered heteroaryl, or C6-C14 aryl, or R _b and R _c together with the N atom may form a heterocycle; r _e can independently represent hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C3-C6 cycloalkenyl, C2-C6 alkynyl, 3-8 membered heterocyclyl, 5-14 membered heteroaryl, or C6-C14 aryl. Typical substituents described above, such as alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocyclyl, heteroaryl or aryl groups and their corresponding substituents and subunits may be optionally substituted, wherein the alkyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl or aryl groups have the definitions described above.

As used herein, the term "substituted" refers to a compound in which any one or more hydrogens on the designated atom are replaced with a selection from the group consisting of the indicated substituents, provided that the normal valency of the designated atom is not exceeded, and the resulting compound from the substitution is stable, i.e., can be isolated, characterized, and subjected to a biological activity test.

Unless otherwise indicated, it is assumed that any heteroatom in an underfilling state has sufficient hydrogen atoms to complement its valence.

As used herein, the term "blend (STATISTIC COPLYMER)" or "random" refers to a polymer formed by the simultaneous polymerization of two or more monomers, at least one of which is of an amino or hydroxyl structure in the side chain, and which is irregularly linked.

As used herein, the term "block-type" refers to a polymer formed by the sequential polymerization of two or more monomers, at least one of which is of a pendant amino or hydroxyl structure, formed from the attachment of different segments.

The structural formula or name of the monomer given in the present invention may only give a specific configuration or not give a specific configuration, and the monomer may also include all other configurations corresponding to the given configuration.

Oxazoline polymers of the invention

The oxazoline polymer of the present invention includes an oxazoline polymer having an amino group in a side chain and/or an oxazoline polymer having a hydroxyl group in a side chain. The oxazoline polymer with the side chain containing amino groups comprises an oxazoline polymer with the side chain amino groups replaced; similarly, the oxazoline polymer having a hydroxyl group in a side chain comprises an oxazoline polymer having a hydroxyl group in a side chain substituted. The oxazoline polymer of the invention also comprises an oxazoline polymer derivative, wherein the derivative refers to that amino groups of a side chain of the polymer are changed into other functional groups such as guanidine groups and the like, or the amino groups contained in the side chain are connected with other molecules such as drug molecules, fluorescent small molecules, protecting groups and the like through chemical reaction; the ends of the polymer are chemically modified, such as by attachment of fluorescent molecules, or drug molecules.

The oxazoline polymer of the invention comprises one or more of the repeating units represented by the formula E ₁ and/or the formula E ₂,

And optionally comprise/>And/or/>One or more of the following. Wherein repeating unit E ₁ and/or formula E ₂ may optionally be linked to repeating units B and/or C.

1. Oxazoline polymer with amino group in side chain

The oxazoline polymer with the side chain containing amino groups refers to a polymer containing one or more repeat units of the formula E ₂; other repeating units B and/or C may also be included.

An exemplary oxazoline polymer having an amino group in a side chain has a structure represented by formula II:

Preferably, the oxazoline polymer having an amino group in a side chain as shown in formula II has a structure as shown in formula V:

preferably, the oxazoline polymer having an amino group in a side chain as shown in formula II has a structure as shown in formula VII:

Preferably, the oxazoline polymer having an amino group in a side chain as shown in formula II has a structure as shown in formula VIII:

in the above-mentioned formulae, the first and second light-emitting elements,

Wherein R ₆ is defined as a structural part other than carboxyl in the molecular structure of a natural amino acid or a non-natural amino acid; n, x, y, z, A, m, m', R ₁、R'₁、R₂、R_a and R _b are as defined above; x, y, z are calculated as the number of repeat units in a side chain divided by the total number of repeat units, e.g., the percent of amino groups in the side chain is the number of units containing amino groups in the side chain divided by the total number of units containing amino groups in the side chain and other functional groups in the side chain.

Preferably, the natural amino acid is selected from the group consisting of: glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartic acid, asparagine, glutamic acid, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine and the like, and derivatives derived from the above amino acids; the unnatural amino acids include beta-amino acids, gamma-amino acids, delta-amino acids, epsilon-amino acids.

Preferably, the derivative derived from the above amino acid is a derivative in which a carboxylic acid group on the amino acid is esterified (e.g., benzyl-esterified, t-butyl-esterified, methyl-esterified, etc.), a derivative in which a hydrogen atom on an amino group on the amino acid is substituted (e.g., substituted with t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), fluorenylmethoxycarbonyl (Fmoc), etc.), a derivative in which a hydrogen atom on a hydroxyl group on the amino acid is substituted (e.g., substituted with t-butyl (tBu)), a derivative in which a hydrogen atom on a free mercapto group on the amino acid is substituted (e.g., substituted with trityl (Trt), benzyl ester, etc.).

Preferably, the amino acid structure is L, D or DL form.

Preferably, the amino or guanidino functional group in the oxazoline polymer structure is in the form of a hydrochloride, hydrobromide, formate, acetate, or trifluoroacetate salt.

Preferably, the amino groups in the oxazoline polymer structure are converted to guanidino functional groups.

Preferably, the amino groups in the polymer structure are protected by protecting groups selected from the group consisting of: t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), fluorenylmethoxycarbonyl (Fmoc), phthaloyl (Pht), acetyl (Ac), trifluoroacetyl (Tfa), benzyl (Bn), triphenylmethyl (Tr).

Preferably, the oxazoline polymer structure is a ternary, quaternary and higher blend or block copolymer.

Preferably, the terminal end of the oxazoline polymer structure is converted into other functional groups such as hydroxyl, amino, ester, etc.

Preferably, after the end of the oxazoline polymer structure is converted into other functional groups such as hydroxyl, amino, ester group and the like, the end of the oxazoline polymer structure is continuously modified by active molecules such as other dye molecules, drug molecules and the like to obtain the oxazoline polymer with the end functionalized.

2. The oxazoline polymer is an oxazoline polymer with a side chain containing hydroxyl

The oxazoline polymer with the side chain containing hydroxyl is a polymer containing one or more repeat units of the formula E ₁; one or more repeating units B and/or C may also be included.

An exemplary oxazoline polymer having an amino group in a side chain has a structure as shown in formula I:

preferably, the oxazoline polymer with a side chain containing hydroxyl groups of formula I has a structure as shown in IV:

/>

Wherein n, x, y, z, m, m', R ₁、R'₁ and R ₂ are as defined above.

3. Oxazoline polymer with amino and hydroxyl groups in side chains

The oxazoline polymer with the side chain containing amino and hydroxyl is a polymer containing repeating units of the formulas E ₁ and E ₁; one or more repeating units B and/or C may also be included.

An exemplary oxazoline polymer having amino and hydroxyl groups in its side chains has a structure as shown in formula III:

Preferably, the oxazoline polymer with side chains containing amino groups and hydroxyl groups, shown in formula III, has a structure shown in formula VI or formula IX:

Wherein, 0% < w <100%; x ₁ is more than or equal to 0% and less than 100%; w and x1 are calculated in the same way as x, y, z.

M, m', n, x, y, z, R ₁、R'₁、R₂、R_a、R_b and R ₆ are as defined above.

The polymer monomer of the invention

The polymer monomers of the present invention include monomers of formula E '₁ and/or E' ₂, and optionally include monomers of formula B 'and/or formula C',

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Preferably, the E '₂ oxazoline monomer has a structure as shown in formula E' ₂:

Preferably, the oxazoline monomer of formula E' ₁ has a structure as shown in formula E "₁:

The definitions of ,R'₁、m'、q'、L₁'、R₁、m、q、L₁、R_a、R_b、P₁、P₂ and R ₇ in the formulae above are as described above.

The synthesis method of the oxazoline monomer with the side chain hydroxyl of the formula E' ₁ comprises the following specific steps:

(2) Reacting compound 2 with a base in a second inert solvent to give a monomer of formula E "₁; wherein m and P ₂、R₁ are as defined above.

X is a leaving group, for example: br, cl, I, OTs, OMs, preferably Cl.

The substitution means substitution with a substituent selected from the group consisting of: halogen, hydroxy, amino, C ₁-C₆ alkyl, C ₁-C₆ alkoxy.

Preferably, the base is selected from the group consisting of: sodium hydroxide, potassium hydroxide, triethylamine, or N, N-diisopropylethylamine, or a combination thereof; sodium hydroxide is preferred.

Preferably, the reaction temperature is selected from the following ranges: 25-100 ℃; preferably 70 ℃.

Preferably, the preparation method of the monomer of the formula IX further comprises the steps of:

(1) Reacting compound 1 with H ₂N(CH₂)₂ X in a first inert solvent to give compound 2; r ₆、P₂、X、n₁ is as defined above.

Preferably, in the step (1), the reaction temperature is 0-70 ℃; preferably, it is from 0℃to 25 ℃.

Preferably, in the step (1), the reaction time is 6 hours to 1 day.

Preferably, in the step (1), H ₂N(CH₂)₂ X is in the form of its hydrochloride, hydrobromide.

Preferably, in step (1), DIEA is replaced with Et ₃ N.

Preferably, the step (1) is to dissolve the compound 1 and DIEA in a first inert solvent, add EDCI and HOBT under ice bath, stir and react for 30min, and then add H ₂N(CH₂)₂ X; after the reaction at room temperature for 8 hours, the reaction mixture is washed twice with deionized water and 5% citric acid aqueous solution respectively, and the organic phase is dried and concentrated, and then is separated and purified by a column to obtain the compound 2.

Preferably, in the step (1), the first inert solvent is independently selected from the group consisting of: DCM, DMF, THF, DMAc; DCM is preferred.

Preferably, the step (2) is to dissolve the compound 2 obtained in the step (1) in a second inert solvent, add alkali for reaction, and protect the compound with inert gas at 70 ℃; after the reaction, the solvent is removed, dichloromethane is added for dissolution, deionized water is used for washing for 2 times, and the organic phase is dried and concentrated, distilled and purified under reduced pressure to obtain a monomer of a formula E' ₁.

In another preferred embodiment, in the step (2), the second inert solvent is independently selected from the group consisting of: methanol, ethanol, acetonitrile, or a combination thereof; preferably selected from: ethanol.

In another preferred embodiment, in the step (2), the reaction is performed under the protection of inert gas; the inert gas is preferably nitrogen, argon, or a combination thereof.

Preparation method of oxazoline polymer of the invention

The oxazoline polymer is prepared by the preparation method of the polymer.

Preparation of homopolymer: in an organic solvent, an oxazoline monomer is subjected to polymerization reaction in the presence of an initiator, so that an oxazoline homopolymer with an amino group in a side chain is obtained.

Preparation of the copolymer: in an organic solvent, in the presence of an initiator, firstly, polymerizing an oxazoline monomer; after the polymerization reaction is finished, adding another oxazoline monomer for polymerization reaction, and so on, when the number of the oxazoline monomers is more, sequentially adding various monomers for polymerization reaction, thereby forming a block type oxazoline copolymer; when the oxazoline monomers are two or more, mixing the two or more oxazoline monomers in an organic solvent, and then carrying out polymerization reaction in the presence of an initiator to form the blend type oxazoline copolymer.

1. Method for preparing oxazoline polymer with side chain containing amino

In an organic solvent, under the protection of inert gas and under the heating condition, the oxazoline monomer represented by the formula E ' ₂ and the optional formula B ' and/or the formula C ' monomer are subjected to polymerization reaction in the presence of an initiator, so that the oxazoline polymer with the side chain containing amino is obtained.

Preferably the preparation method comprises the steps of:

In the protection of an organic solvent and inert gas, under the existence of an initiator and heating conditions, polymerizing at least one oxazoline monomer with side chain amino protection, thereby obtaining an oxazoline polymer with side chain amino protection; removing the amino protecting group to obtain a 2-oxazoline polymer with an amino side chain; after the guanidine group is functionalized, the oxazoline polymer with the side chain guanidine group is obtained.

Preferably, the organic solvent is selected from the group consisting of: DMAc, DMF, or a combination thereof; DMAc is preferred.

Preferably, the inert gas is selected from the group consisting of: n ₂, ar; ar is preferred.

Preferably, the initiator is selected from the group consisting of: meOTf, bzOTf; preferably MeOTf.

Preferably, the heating conditions result in a temperature range of 80 ℃ to 140 ℃; preferably 120 ℃.

Preferably, the polymerization reaction time is 2h-8h; preferably 6h.

Preferably, the oxazoline copolymer with side chain amino groups is a polymer obtained by mixing two or more monomers in a set proportion and then copolymerizing.

Preferably, when the oxazoline monomers are two or more, the method comprises the steps of:

in an organic solvent, in the presence of an initiator, firstly, polymerizing an oxazoline monomer;

After the polymerization reaction is finished, adding another oxazoline monomer for polymerization reaction, thereby forming a block oxazoline copolymer.

By analogy, when the oxazoline monomers are more, various monomers can be sequentially added to perform polymerization reaction.

Preferably, when the oxazoline monomers are two or more, the method comprises the steps of: in an organic solvent, two or more oxazoline monomers are mixed and then subjected to polymerization reaction in the presence of an initiator, so that an oxazoline copolymer is formed.

It is understood that one, two, three or more oxazoline monomers are polymerized and that at least one of the monomer side chains is an amino group or a protected amino group.

2. Preparation method of oxazoline polymer with side chain containing hydroxyl

Preferably the preparation method comprises the steps of:

In the protection of an organic solvent and inert gas, under the existence of an initiator and heating conditions, polymerizing at least one oxazoline monomer with side chain hydroxyl protection, thereby obtaining an oxazoline polymer with side chain hydroxyl protection; removing the hydroxyl protecting group to obtain the oxazoline polymer with the side chain of hydroxyl.

Preferably, the organic solvent is selected from the group consisting of: DMAc, DMF, meCN, phCN, or a combination thereof; DMAc is preferred.

Preferably, the inert gas is selected from the group consisting of: n ₂ and Ar; ar is preferred.

Preferably, the initiator is selected from the group consisting of: meOTf, bzOTf, CH ₃(CH₂)₅Br、CH₃(CH₂)₅ Ots, or a combination thereof; preferably MeOTf.

Preferably, the heating conditions result in a temperature range of 80 ℃ to 140 ℃; preferably from 80℃to 120 ℃.

Preferably, the polymerization reaction time is 2h-8h; preferably 6h.

Preferably, the oxazoline copolymer with side chain hydroxyl groups is a polymer obtained by mixing two or more monomers in a set proportion and then copolymerizing.

Preferably, when the oxazoline monomers are two or more, the method comprises the steps of: in an organic solvent, two or more oxazoline monomers are mixed and then subjected to polymerization reaction in the presence of an initiator, so that a blend type oxazoline copolymer is formed.

It is understood that one, two, three or more oxazoline monomers are polymerized and at least one of the monomer side chains is a hydroxyl group or a protected hydroxyl group, and the additional monomer may be an amino-protected oxazoline monomer or any other polymerizable monomer.

3. Method for preparing oxazoline polymer with side chain containing amino and hydroxyl

The preparation method comprises the following steps: in an organic solvent, under the existence of an initiator and heating conditions, carrying out polymerization reaction on an oxazoline monomer represented by a formula E '₂, an oxazoline monomer represented by a formula E' ₁ and optionally oxazoline monomers with other structures, so as to obtain an oxazoline polymer with amino groups and hydroxyl groups on side chains; wherein E' ₂、E'₁ is defined as described above.

Preferably, optionally the oxazoline monomer of other structure is a monomer of formula B 'or formula C'.

In a preferred embodiment, one or more oxazoline monomers represented by the formula E "₂ and one or more E" ₁ and optionally B 'and/or C' monomers are polymerized in an organic solvent in the presence of an initiator and with heat to give an oxazoline polymer having amino-protected and hydroxyl-protected side chains; removing the protecting group to obtain oxazoline polymer with side chain amino and hydroxyl; optionally, the amino groups in the oxazoline polymer from which the amino groups and the hydroxyl groups are obtained are functionalized towards the guanidine groups to obtain the oxazoline polymer from which the side chain guanidine groups and the hydroxyl groups are obtained.

Preferably, the oxazoline copolymer with the side chain containing amino and hydroxyl is a polymer obtained by mixing two or more monomers in a set proportion and then copolymerizing.

In an organic solvent, under the existence of an initiator and heating conditions, firstly, carrying out polymerization reaction on an oxazoline monomer (amino or hydroxyl side chain);

After the polymerization reaction is finished, adding another oxazoline monomer (hydroxyl or amino side chain) to perform the polymerization reaction, thereby forming a block oxazoline copolymer;

Preferably, when the oxazoline monomers are two or more, the method comprises the steps of: in an organic solvent, oxazoline with amino and hydroxyl groups in side chains or any one or more other polymerizable monomers (preferably B 'and/or C') are mixed and then subjected to polymerization reaction in the presence of an initiator, so that a blended oxazoline copolymer is formed.

It will be appreciated that two, three or more oxazoline monomers are polymerized and that at least two of the monomer side chains are amino or hydroxy or protected amino or hydroxy groups, respectively, and that the additional monomer may be any other polymerizable monomer.

Use of the polymers according to the invention

1. Oxazoline monomer with amino group in side chain

The oxazoline monomer with the side chain containing amino can be used in the biomedical fields such as antibiosis, anti-tumor, tissue engineering, drug and gene delivery, drug modification, cell adhesion, self-assembly material and the like.

Preferably, the antimicrobial material is in the form of a solution, a surface coating.

2. Oxazoline polymer with side chain containing hydroxyl

The application of the oxazoline polymer with side chain hydroxyl is used in the fields of surface antifouling, protein modification and protection, cell protection, tissue and organ freeze protection, drug modification and the like.

Preferably, the oxazoline polymer of side chain hydroxyl groups is used for a surface antifouling coating.

3. Oxazoline polymer with side chain containing amino and hydroxyl

The oxazoline polymer with the side chain containing amino and hydroxyl can be used in the biomedical fields of antibiosis, anti-tumor, tissue engineering, drug and gene delivery, drug modification, cell adhesion, self-assembly materials, surface antifouling, cell freezing, drug modification and the like.

The invention has the main advantages that:

1. The invention provides a structure and a preparation method of an oxazoline polymer with amino-containing side chains and derivatives thereof, and the oxazoline polymer with amino-containing side chains and derivatives thereof can be used for polymer function research of biological medicines such as solution antibiosis, surface antibiosis, drug synergistic antibiosis, drug delivery, drug modification, cell adhesion, gene delivery, self-assembly materials, anti-tumor, cell adhesion and the like. Compared with the traditional oxazoline polymer based on methyl or ethyl oxazoline as a hydrophilic structure, the invention expands the application range of the oxazoline polymer system in the field of biological medicine.

2. The invention provides a structure and a preparation method of a polymer of an oxazoline monomer with a side chain containing hydroxyl and derivative thereof, and the oxazoline polymer with the side chain hydroxyl and derivative thereof can be used in the fields of surface antifouling, protein modification and protection, cell protection, tissue and organ cryoprotection, drug modification and the like.

3. The oxazoline polymer with the side chain containing the amino group and the hydroxyl group has the functions of the oxazoline polymer with the side chain containing the amino group and the hydroxyl group

The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Percentages and parts are weight percentages and parts unless otherwise indicated. The experimental materials and reagents used in the following examples were obtained from commercial sources unless otherwise specified.

Examples

Preparation method of oxazoline monomer with side chain amino

Preparation of N-Boc-2- (aminomethyl) oxazoline monomer

N-Boc glycine (3.5 g,20 mmol) was dissolved in 100mL of methylene chloride, EDCI (5.8 g,30 mmol), HOBT (2.7 g,30 mmol) and triethylamine (8.3 mL,60 mmol) were sequentially added under ice bath, after stirring for 30 minutes, 2-chloroethylamine hydrochloride (3.0 g,26 mmol) was added, after stirring at room temperature for 8 hours, the reaction mixture was washed twice with 30mL of deionized water, 30mL of 5% aqueous citric acid solution, the organic phase was collected, dried over anhydrous magnesium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography to give intermediate (3.3 g, yield 70%).

Sodium hydroxide (0.8 g,20 mmol) was dissolved in 35mL ethanol in a dry single-necked flask, then intermediate (2.7 g,10 mmol) was added and heated to reflux under nitrogen protection for 2h. The reaction solution was concentrated, and 30mL of methylene chloride was added for dissolution, extracted twice with deionized water, and finally the organic phase was dried, concentrated, and then recrystallized to give needle-like N-Boc-2- (aminomethyl) oxazoline monomer (1.3 g, yield 65.5%).

Preparation of N-Boc-2- (aminopropyl) oxazoline monomer

The experimental procedure was identical to 1 except that N-Boc glycine (3.5 g,20 mmol) was replaced with N-Boc aminobutyric acid (4.1 g,20 mmol).

3.Preparation of 2- (isobutyl) oxazoline monomer

Isopentanonitrile (1.6 g,20 mmol), ethanolamine (1.2 g,20 mmol) and zinc acetate (0.2 g,1 mmol) were added to a dry single-necked flask, mixed and stirred, reacted at 140℃for 16h under nitrogen protection, cooled to room temperature after the reaction was completed, then 100mL of methylene chloride was added to dissolve, the organic phase was washed three times with 50mL of deionized water, the organic phase was dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. To the concentrated liquid, 1g of calcium hydride was added, and finally, purification by distillation under reduced pressure was performed to obtain 2- (isobutyl) oxazoline monomer (1.5 mg, yield 58%) as an oil.

4.Preparation of 2- (butyl) oxazoline monomer

The experimental procedure was identical to 3, except that isovaleronitrile (1.6 g,20 mmol) was replaced with n-valeronitrile (1.6 g,20 mmol).

5.2 Preparation of the (naphthylmethyl) oxazoline monomer

The experimental procedure was identical to 1, except that naphthalene acetic acid (3.7 g,20 mmol) was used instead of N-Boc glycine (3.5 g,20 mmol).

Example 12 Synthesis of an aminomethyloxazoline homopolymer

In a glove box protected by nitrogen, monomer N-Boc-2- (aminomethyl) oxazoline and initiator methyl triflate are respectively dissolved in dry N, N-2-methylacetamide, 100 mu L of initiator methyl triflate solution with the concentration of 0.2M and 2mL of monomer N-Boc-2- (aminomethyl) oxazoline with the concentration of 0.2M are added into a flask to be mixed, and then the mixture is stirred and reacted for 6h at 120 ℃;

After the polymerization was completed, the reaction mixture was cooled to room temperature, and the above reaction mixture was poured into cold petroleum ether (45 mL), and the white floccule precipitated was collected by centrifugation, dried in an air stream, redissolved in tetrahydrofuran (1.5 mL), and precipitated by adding a large amount of cold petroleum ether. This dissolution-precipitation procedure was repeated three times in total to give 72.6mg (yield 90%) of a homopolymer of N-Boc-2- (aminomethyl) oxazoline.

The molecular weight mn=4100 and the molecular weight distribution PDI (Mw/Mn) =1.22 of the polymer obtained by MeOTf initiating polymerization were identified by Gel Permeation Chromatography (GPC).

Then adding 2mL of trifluoroacetic acid into the pumped polymer, slightly shaking at room temperature for about 2 hours, blowing off excessive trifluoroacetic acid, dissolving the obtained viscous liquid into 0.5mL of methanol, adding 45mL of frozen diethyl ether to separate out white precipitate, and repeating the dissolving-precipitating process for three times to obtain the oxazoline homopolymer with side chain amino groups. The deprotected polymer was again dissolved in 5mL of ultrapure water, and filtered and lyophilized for subsequent bioactivity testing.

Comparative example 1 polymerization of 2-aminomethyloxazoline with acetonitrile as solvent

In a glove box protected by nitrogen, respectively dissolving monomer N-Boc-2- (aminomethyl) oxazoline and initiator methyl triflate in a dry acetonitrile solvent, adding 100 mu L of initiator methyl triflate solution with the concentration of 0.2M and 2mL of monomer N-Boc-2- (aminomethyl) oxazoline with the concentration of 0.2M into a flask, mixing, and stirring under the condition of heating and refluxing for reaction for 6 hours; the analysis by thin layer chromatography showed that most of the monomer materials did not participate in the polymerization reaction, while cooling the reaction solution to room temperature, and pouring the above reaction mixture into cold petroleum ether (45 mL), collecting a small amount of white floccules precipitated by centrifugation, drying in a gas stream, redissolving in tetrahydrofuran (1.5 mL), and adding a large amount of cold petroleum ether for precipitation. After three total repetitions of this dissolution-precipitation process, no white solid was collected.

This comparative example 1 shows that it is difficult to perform the polymerization reaction when acetonitrile is used as a solvent (reaction temperature is about 80 ℃ (reflux temperature)).

EXAMPLE 2 Synthesis of guanidine-derivatized oxazoline homopolymers

In a glove box protected by nitrogen, monomer N-Boc-2- (aminopropyl) oxazoline and initiator methyl triflate (MeOTf) are respectively dissolved in dry N, N-2-methylacetamide, 100 mu L of initiator methyl triflate solution with the concentration of 0.2M and 2mL of monomer N-Boc-2- (aminopropyl) oxazoline with the concentration of 0.2M are added into a flask and mixed, and then stirred at 120 ℃ for reaction for 6 hours;

After the polymerization was completed, the reaction mixture was cooled to room temperature, and the above reaction mixture was poured into cold petroleum ether (45 mL), and the white floccule precipitated was collected by centrifugation, dried in an air stream, redissolved in tetrahydrofuran (1.5 mL), and precipitated by adding a large amount of cold petroleum ether. This dissolution-precipitation procedure was repeated three times in total to give 84.1mg (92% yield) of a homopolymer of N-Boc-2- (aminopropyl) oxazoline.

The molecular weight mn=4710 and the molecular weight distribution PDI (Mw/Mn) =1.17 of the polymer obtained by MeOTf initiated polymerization were identified by Gel Permeation Chromatography (GPC).

Then adding 2mL of trifluoroacetic acid into the pumped polymer, slightly shaking at room temperature for about 2 hours, blowing off excessive trifluoroacetic acid, dissolving the obtained viscous liquid into 0.5mL of methanol, adding 45mL of frozen diethyl ether to separate out white precipitate, and repeating the dissolving-precipitating process for three times to obtain the oxazoline homopolymer with side chain amino groups.

Then 50mg of the drained polymer is dissolved in 1.5mL of methanol, 90mg of N, N-diisopropylethylamine and 106mg of 1H-pyrazole-1-carboxamide hydrochloride are added, the mixed solution is stirred for 12 hours under the protection of nitrogen, after the excessive solvent is removed by heating and concentration, the obtained viscous liquid is dissolved in 0.5mL of methanol, 45mL of frozen acetone is added to separate out white precipitate, and the dissolving-precipitating process is repeated three times, so that the oxazoline homopolymer with a side chain guanidino group is obtained. The oxazoline polymer with side chain guanidine groups was again dissolved with 5mL of ultrapure water and filtered and lyophilized for subsequent bioactivity testing.

Example 32 Synthesis of a Polymer blend of (aminomethyl) oxazoline and 2- (isobutyl) oxazoline

In a glove box protected by nitrogen, monomer N-Boc-2- (aminomethyl) oxazoline, monomer 2- (isobutyl) oxazoline and initiator methyl triflate (MeOTf) are respectively dissolved in dry N, N-2-methylacetamide, 100 mu L of initiator methyl triflate solution with the concentration of 0.2M and 0.8mL of monomer N-Boc-2- (aminomethyl) oxazoline with the concentration of 0.2M are added into a flask, mixed and then stirred at 120 ℃ for reaction for 6 hours;

After the polymerization was completed, the reaction mixture was cooled to room temperature, and the above reaction mixture was poured into cold petroleum ether (45 mL), and the white floccule precipitated was collected by centrifugation, dried in an air stream, redissolved in tetrahydrofuran (1.5 mL), and precipitated by adding a large amount of cold petroleum ether. This dissolution-precipitation procedure was repeated three times in total to obtain 57.6mg (yield 92%) of a copolymer of oxazoline with protected side-chain amino groups.

The molecular weight mn=3170 and the molecular weight distribution PDI (Mw/Mn) =1.18 of the polymer obtained by MeOTf-initiated polymerization were identified by Gel Permeation Chromatography (GPC).

Then adding 2mL of trifluoroacetic acid into the pumped polymer, slightly shaking for about 2 hours at room temperature, blowing off excessive trifluoroacetic acid, dissolving the obtained viscous liquid into 0.5mL of methanol, adding 45mL of frozen diethyl ether to separate out white precipitate, and repeating the dissolving-precipitating process for three times, thereby obtaining the amphiphilic oxazoline blend polymer with side chain amino groups. The deprotected polymer was again dissolved in 5mL of ultrapure water, and filtered and lyophilized for subsequent bioactivity testing.

Example 42 Synthesis of a blend Polymer of (aminomethyl) oxazoline and 2- (naphthylmethyl) oxazoline

The experimental procedure was identical to example 3 except that the hydrophobic monomer was replaced with 2- (naphthylmethyl) oxazoline, cooled to room temperature after the polymerization reaction was completed, cold petroleum ether (45 mL) was poured into the reaction mixture, and the precipitated white floccules were collected by centrifugation, dried in an air stream, redissolved in tetrahydrofuran (1.5 mL), and precipitated by adding a large amount of cold petroleum ether. This dissolution-precipitation procedure was repeated three times in total to give 78.2mg (yield 95%) of a copolymer of oxazoline with protected side-chain amino groups.

The molecular weight mn=4290 and the molecular weight distribution PDI (Mw/Mn) =1.17 of the polymer obtained by MeOTf initiated polymerization were identified by Gel Permeation Chromatography (GPC).

Example 52 Synthesis of block copolymers of (aminomethyl) oxazoline and 2- (naphthylmethyl) oxazoline

The experimental procedure was identical to example 4, except that after the reaction of the N-Boc-2- (aminomethyl) oxazoline monomer was completed, the hydrophobic monomer 2- (naphthylmethyl) oxazoline was added, cooled to room temperature after the polymerization of the hydrophobic monomer was completed, cold petroleum ether (45 mL) was poured into the reaction mixture, the precipitated white floccules were collected by centrifugation, dried in air flow, redissolved in tetrahydrofuran (1.5 mL), and a large amount of cold petroleum ether was added for precipitation. This dissolution-precipitation procedure was repeated three times in total to obtain 74.1mg (yield 90%) of a block polymer of oxazoline with protected side-chain amino groups.

The molecular weight mn=4300 and the molecular weight distribution PDI (Mw/Mn) =1.23 of the polymer obtained by MeOTf initiating polymerization were identified by Gel Permeation Chromatography (GPC).

EXAMPLE 6 Synthesis of thiol-functionalized amphiphilic oxazoline Polymer libraries with pendant amino groups

Experimental procedure the experimental procedure is the same as in example 3, except that the hydrophobic monomer is replaced with 2- (butyloxazoline) oxazoline and the polymerization is carried out by adjusting the proportions of the different oxazoline monomers (proportions from 90% of N-Boc-2- (aminomethyl) oxazoline monomers +10% of 2- (butyl) oxazoline to 30% of N-Boc-2- (aminomethyl) oxazoline monomers +70% of 2-butyloxazoline); after the polymerization reaction was completed, 20mg of 3- (tritylthio) propionic acid, 13mg of N, N-diisopropylethylamine and 12mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride were added to continue the reaction for 12 hours, then cooled to room temperature, and the above reaction mixture was poured into cold petroleum ether (45 mL), and the white flocculent precipitated was collected by centrifugation, dried in a gas stream, redissolved in tetrahydrofuran (1.5 mL), and a large amount of cold petroleum ether was added to precipitate. This dissolution-precipitation process was repeated three times in total to obtain a series of oxazoline copolymers with different hydrophilic-hydrophobic ratios (yield 85% -96%) after thiol functionalization.

Then adding 2mL of trifluoroacetic acid and 5% (v/v) of triethylsilane into the pumped polymer, stirring overnight at room temperature, blowing off excessive trifluoroacetic acid by using nitrogen, dissolving the obtained viscous liquid in 0.5mL of methanol, adding 45mL of frozen diethyl ether to separate out white precipitate, and repeating the dissolving-precipitating process for three times, thereby obtaining the oxazoline polymer library with side chain amino groups after mercapto functionalization. The deprotected polymer was again dissolved in 5mL of ultrapure water, and filtered and lyophilized for subsequent bioactivity testing.

It will be appreciated that the oxazoline monomers with protected amino groups in the side chains in the examples can be successfully prepared into oxazoline polymers with amino groups in the side chains using methyl triflate as initiator and under the other polymerization conditions, which breaks the recognition that methyl triflate is traditionally regarded as unsuitable for the initiator of polymerization of oxazoline monomers with protected amino groups in the side chains.

EXAMPLE 7 use of oxazoline Polymer and derivatives of side chain amino groups as antibacterial materials in solution

Minimum Inhibitory Concentration (MIC) test bacteria were cultured overnight with LB liquid medium (Luria-BertaniBroth) in a shaker at 37 ℃ at 150rpm, and bacterial cells obtained by the culture were collected by centrifugation and re-dispersed into MH (Mueller-Hinton Broth) medium, and absorbance at 600nm (OD ₆₀₀) was read with a microplate reader(s) (s. Staphylococcus aureus concentration was about 1.5x10 ⁹ CFU/mL when OD ₆₀₀ =1). The bacterial solution was diluted to 2X 10 ⁵ CFU/mL with MH medium for use. The polymer was diluted in 96-well plates with MH medium at a concentration ranging from 400 to 3.13. Mu.g/mL. Then, 50. Mu.L of diluted bacterial liquid was added to each well to make the total volume of the bacterial liquid and the polymer 100. Mu.L, and the mixture was gently shaken for 10 seconds and allowed to stand for 9 hours in a mold incubator at 37 ℃. OD ₆₀₀ was then read with an enzyme-labeled instrument, and 4 wells of the same 96-well plate were added with MH medium alone as negative control, and 4 wells were added with MH medium and bacterial fluid (without polymer) as positive control. Two replicates were tested at a time and repeated twice at different times. The percentage of bacterial growth per well uses the formulaAnd (5) calculating. The calculated data is then plotted and the MIC value is the lowest concentration of polymer that inhibits bacterial growth.

The polymers were tested for their minimum inhibitory concentration against a variety of bacteria, including methicillin-resistant staphylococcus aureus USA300 (Staphylococcus aureus USA) 300, methicillin-resistant staphylococcus aureus Mu50 (Staphylococcus aureus Mu), bacillus subtilis (Bacillus subtilis BR-151), escherichia coli (ESCHERICHIA COLI JM), pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC ₉₀), multidrug-resistant pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC ₁₅ 442), sulfamethoxazole and tetracycline naturally-resistant pseudomonas aeruginosa (Pseudomonas aeruginosa O1), acinetobacter baumannii (Acinetobacter baumannii ATCC BAA-747). The minimum inhibitory concentrations of the 2-aminomethyloxazoline homopolymer prepared in example 1 against positive bacteria Staphylococcus aureus USA, staphylococcus aureus Mu, and Bacillus subtilis BR-151 were 12.5 μg/mL, and 3.13 μg/mL, respectively; the minimum inhibitory concentrations of the guanidino-functionalized 4-aminopropyl oxazoline homopolymer prepared in example 2 on positive bacteria Staphylococcus aureus USA, staphylococcus aureus Mu, 50 and Bacillus subtilis BR-151 were 100 μg/mL, 100 μg/mL and 6.25 μg/mL, respectively; the lowest inhibitory concentration of the oxazoline polymer with side chain amino group prepared in example 3 was 3.13 μg/mL for negative bacteria pseudomonas aeruginosa Pseudomonas aeruginosa ATCC15442, pseudomonas aeruginosa ATCC ₉₀ 27 and Pseudomonas aeruginosa O1; the minimum inhibitory concentrations for the negative bacteria Acinetobacter baumannii Acinetobacter baumannii ATCC BAA-747 and the E.coli ESCHERICHIA COLI JM were 12.5. Mu.g/mL and 6.25. Mu.g/mL, respectively. The obtained MIC results prove that the oxazoline polymer with side chain amino and the guanylated derivative have stronger antibacterial activity.

EXAMPLE 8 use of oxazoline polymers and derivatives of pendant amino groups as solution antifungal materials

The minimum inhibitory concentration (MFC) of the guanidino-functionalized oxazoline homopolymer prepared in example 2 on Candida albicans K1 (C.albicans K1), cryptococcus neoformans H99 (C.neoformans H99) and Cryptococcus neoformans JEC21 (C.neoformans JEC 21) resulted in 6.25. Mu.g/mL (C.albicans K1), 3.13. Mu.g/mL (C.neoformans H99) and 0.78. Mu.g/mL (C.neoformans JEC 21); the minimum inhibitory concentration (MFC) of 2-aminomethyloxazoline and 2-naphthylmethyl oxazoline polymer blend prepared in example 4 on Candida albicans K1, cryptococcus neoformans H99 and Cryptococcus neoformans JEC21 was 6.25. Mu.g/mL (C.albicans K1), 3.13. Mu.g/mL (C.neoformans H99) and 0.78. Mu.g/mL (C.neoformans JEC 21). The obtained MFC result proves that the oxazoline polymer and the derivative with the side chain amino groups have stronger antifungal activity.

EXAMPLE 9 use of oxazoline Polymer with pendant amino groups as surface coating antibacterial Material

The polymer synthesis was carried out as described in example 6, except that the deprotected thiol-terminated oxazoline polymer was grafted onto the surface of the gold plate, and the surface sterilization test was carried out by culturing the bacteria overnight with LB liquid medium (Luria-Bertani Broth) in a shaking table at 37℃at 150 rpm. After the completion of the culture, 7.5mL of the bacterial liquid was taken out from the conical flask, centrifuged at 4000rpm for 5min to collect the bacteria, re-dispersed in PBS, re-centrifuged, and the bacterial liquid was collected after repeating the centrifugation of the PBS dispersion three times, and the number of colonies was quantified by reading absorbance at 600nm (OD ₆₀₀) with an ELISA reader. The bacterial liquid is diluted to 1X 10 ⁵ CFU/mL by PBS for standby. The prepared polymeric antimicrobial surface was placed in a 24-well plate with PBS as a control. And (3) adding 80 mu L of the bacterial liquid with the concentration on the surface of a polymer gold slice, wherein 80 mu L of the bacterial liquid is directly added into a pore plate to serve as a blank control, PBS is added into the blank pore plate to control humidity, the mixture is subjected to static culture in a mould incubator at 37 ℃ for 2.5 hours, the pore plate is taken out, 1920 mu L of PBS is added into the pore plate to dilute, the mixture is subjected to ultrasonic treatment for 3min, the mixture is uniformly mixed for 2min under a mixer, 30 mu L of the mixture is taken out by a pipette gun and is coated on LB agar medium, and the mixture is placed in the mould incubator at 37 ℃ to be cultured. After colony counting, surface antibacterial activity analysis was performed, and the experimental group was designated as C _{Sample of} and the blank control was designated as C _{Blank space}. The antibacterial activity (bacterial kill rate) of the substrate surface was calculated from the following formula:

Thiol-functionalized oxazoline-co-polymer with pendant amino groups (preferably 90% of N-Boc-2- (aminomethyl) oxazoline +10% of 2- (butyl) oxazoline) was tested for surface sterilization against Methicillin-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA, methicillin-resistant staphylococcus aureus). Experimental results prove that the sterilization rate of the surface of the blended oxazoline polymer to MRSA can reach 99.9%, and the surface sterilization effect is excellent.

EXAMPLE 10 use of oxazoline Polymer with side chain amino groups as cell adhesion Material

The polymer prepared in example 6 was grafted onto the surface of a glass substrate by using 3-aminopropyl triethoxysilane as a glass surface amino modifier to modify and clean the surface-activated glass sheet, then using PEG to modify the amination slide, and finally grafting an amino acid polymer and a positive control polypeptide (RGD). Trypsin digestion and cell collection are carried out in a centrifuge tube, and the cell density is adjusted to 8 multiplied by 10 ⁴ cells/mL; seeding cells into pores of the amino acid polymer surface; the polymer surface was placed in a petri dish and incubated in an incubator at 37 ℃. After incubating the cells for 2 hours, observing the states of adherence, spreading, agglomeration and the like of the cells on the surface of the polymer under an inverted microscope; the surface of the polymer attached with the cells was then immersed in the medium for continuous culture for 24 to 48 hours, and the morphology of the cell adhesion growth on the amino acid polymer surface was observed for a plurality of areas using an inverted fluorescence microscope, and the coverage area (%) of the cell surface was calculated. Experimental results show that on the glass surface of the grafted oxazoline polymer, the mouse embryonic fibroblasts (NIH 3T 3) show different adhesion effects at 48h, the preferred polymer is 40% N-Boc-2- (aminomethyl) oxazoline+60% 2- (butyl) oxazoline, the polymer shows similar cell adhesion effects as the positive control RGD, and the number of cells grown on the surface grafted with the polymer is 75% of the number of cells on the surface of the positive control RGD by counting. Cell adhesion is a key step in the action of cells and materials in tissue engineering, and cells can perform a series of actions such as subsequent proliferation, migration, differentiation and the like only through adhesion, so that supporting cell adhesion is an essential property of biological materials in the application of tissue engineering.

EXAMPLE 11 application of oxazoline Polymer with side chain amino group as antitumor Material

Cytotoxicity test (MTT cell proliferation assay) NCI-H460 cells, U87 cells, and B16 cells, each having a density of 3X 10 ⁴, were seeded in 96-well plates at a volume of 100. Mu.L per well, respectively, by the following method. Cells were cultured at 37℃for 24 hours. After removal of the old medium, medium containing different concentrations of amino acid polymer was added, and three duplicate wells were set for each concentration. After culturing the cells at 37℃for 24 hours, 10. Mu.L of MTT solution (5 mg/mL in PBS) was added to each well, and incubation was continued for 4 hours, and the culture was terminated. The culture supernatant was carefully aspirated off from the wells, DMSO (150. Mu.L) was added to each well, and the mixture was shaken on a shaker for 10 minutes to allow the crystals to dissolve well. Cells treated without any amino acid polymer were included as controls on the same 96-well plate, and a blank without seeding cells with DMSO alone. The 570nm wavelength was selected, the light absorption value (OD value) of each well was measured on a microplate reader, and the cell viability was calculated: % cell viability= (OD ^Polymer–OD^{Blank space})/(OD^Control–OD^{Blank space}) x 100. On this basis, the cell viability was plotted as a function of the amino acid polymer concentration, and the lowest amino acid concentration (IC ₅₀) that resulted in 50% mammalian cell death was obtained from the plot.

The oxazoline polymer blend polymer of side chain amino group prepared in example 3 (the ratio from 40% of N-Boc-2- (aminomethyl) oxazoline to 60% of 2- (isobutyl) oxazoline) was tested for cytotoxicity against various tumor cells (NCI-H460 lung cancer cell, U87 glioma cell, B16 melanoma cell), and the experimental result shows that the oxazoline polymer has an IC ₅₀ of 25 μg/mL against NCI-H460 lung cancer cell, exhibiting good antitumor effect.

EXAMPLE 12 use of oxazoline Polymer with pendant amino groups as self-assembled Material

The oxazoline block copolymer with side chain amino groups prepared in example 5 is preferable, and the self-assembled structure of the polymer is prepared by dissolving 1mg of the deprotected oxazoline polymer in ultrapure water of corresponding volume to prepare a solution of 0.2mg/mL or 0.5mg/mL, and standing for 12 hours after stirring the solution at a medium speed at 390rpm for 2 hours. The self-assembled solution was filtered using a 0.8 μm filter head and subjected to DLS testing.

The self-assembled samples were tested for particle size and dispersibility using Dynamic Light Scattering (DLS). Samples were placed in PS cuvettes, each sample was tested at a volume of about 1.5mL, each sample was repeated three times at a temperature of 25 ℃ and a test angle set at 90 degrees. Data processing uses a cumulative analysis of the experimental correlation functions and uses Stokes-Einstein equations to calculate the diffusion coefficients.

The DLS experiment result shows that the particle size of the self-assembled structure formed by the oxazoline polymer in water is 50-100nm, and the dispersity PD is 0.2-0.4.

EXAMPLE 13 use of oxazoline Polymer with side chain amino group as drug delivery Material

The oxazoline block copolymer with side chain amino groups prepared in example 5 is preferable, and the polymer as a drug delivery material is prepared by dissolving the preferable oxazoline block copolymer (40 mg/mL) and amphotericin B (10 mg/mL) in DMSO, taking 200. Mu.L of the polymer solution, dropwise adding (1 drop/2 s) to ultra-pure water stirred (360 r-390 r), continuing to stir for 2h in a dark place, standing for 8h in a dark place, and filtering with a 0.8 μm filter head. Dialysis was performed with 3500 molecular weight dialysis bag for 4 hours, with 30 minutes for one liquid change (10% DMSO in ultrapure water), and the resulting liquid was stored for a long period of time at 4 ℃.

The DLS experimental result shows that the particle size of the self-assembled structure formed in water after the oxazoline polymer with the side chain amino group is used for coating the drug amphotericin B is 50-100nm, and the dispersity PD is 0.2-0.4.

The hemolysis and antifungal activity of the amphotericin B-entrapped material are tested, and the hemolysis of the amphotericin B-entrapped material on human mammal erythrocytes is HC ₅₀ & gt 200 mug/mL, and the antifungal activity on fungi C.albicans K1 is respectively MFC=12.5 mug/mL; compared with the single drug amphotericin B which has the hemolysis of human mammal erythrocytes of HC ₅₀ =15.6 μg/mL, the antifungal activity on fungi c.albicans K1 has mfc=1.56 μg/mL respectively; experimental results show that the oxazoline polymer with side chain amino groups as a drug delivery material maintains a stable self-assembled structure, and simultaneously reduces the hemolytic toxicity of the drug amphotericin B.

Example 14 application of oxazoline Polymer with side chain amino group as pharmaceutical synergistic antibacterial Material

The oxazoline polymer with side chain amino groups prepared in example 3 is preferable, and the oxazoline polymer is used for a synergistic antibacterial test by dispersing bacteria in 10mL of LB medium, placing the medium in a shaking table at 37 ℃ for 10-11 hours at a rotation speed of 150rpm, centrifuging a bacterial medium solution obtained by overnight culture at a rotation speed of 4000rpm, pouring out supernatant obtained by centrifugation, adding 5mL of MH medium to bacteria obtained by centrifugal precipitation, and shaking to redisperse the bacteria in the MH medium. OD ₆₀₀ was read with an ELISA reader and then diluted with MH medium to determine a concentration of 5X 10 ⁵ CFU/mL. The polymer solution was diluted to the appropriate concentration by a two-fold stepwise dilution method in a 96-well plate, and the volume of the polymer solution in each well was 40. Mu.L. The antibiotic solution was diluted laterally to the appropriate concentration by two-fold stepwise dilution in another 96-well plate and 40 μl of antibiotic solution was transferred line-by-line to a 96-well test plate. Then 20 mu L of bacterial liquid is added into a 96-well test plate, so that the final concentration of the bacterial liquid is 1 multiplied by 10 ⁵ CFU/mL, the final concentration of the polymer and the antibiotic is 200 mu g/mL-1.56 mu g/mL, the wells of 100 mu L of MH culture medium are used as negative control, and the wells of 100 mu L of bacterial liquid with the bacterial concentration of 1 multiplied by 10 ⁵ CFU/mL are used as positive control. Placing the 96-well plate on a desktop, horizontally and slightly shaking for 10 seconds, placing the 96-well plate in a mould incubator at 37 ℃ for static culture, reading OD ₆₀₀ by using an enzyme-labeled instrument after 9 hours, and calculating the bacterial growth rate:

% bacterial growth= (OD ^{Experimental group}-OD^{negative control})/(OD^{Positive control}-OD^{negative control}) x 100

The MIC (Minimum inhibition cconcentration) value is the minimum concentration at which the polymer causes inhibition of bacterial growth. The FICI (Fractional Inhibitory Concentration Index) value is used for representing the antibacterial effect of the combination of two substances with different concentrations, and calculating the fractional inhibition concentration index:

FICI＝(MIC^{Polymer combination}/MIC^{polymer for single use})+(MIC^{Antibiotic combinations}/MIC^{Antibiotic single use})

On the basis, FICI less than or equal to 0.5 is the synergistic effect of the two substances.

The oxazoline polymer prepared in example 3 was tested for synergistic antibacterial activity with antibiotic rifampin against various gram-negative bacteria (including pseudomonas aeruginosa p.aerocinosa O1; acinetobacter baumannii a.baumannii atccbaa747; escherichia coli e.coll ATCCBAA 747), and the experimental result showed that the oxazoline polymer prepared in example 3 had a FICI of 0.51 against bacteria p.aerocinosa O1 after rifampin was used; the FICI for bacterium baumanniiATCCBAA and 747 is 0.25 and the FICI for bacterium E.coli ATCCBAA and 747 is 0.37, which indicates that the oxazoline polymer with side chain amino group and rifampicin have excellent synergistic antibacterial effect against Acinetobacter baumannii and Escherichia coli and additive antibacterial effect against Pseudomonas aeruginosa.

EXAMPLE 15 Synthesis of oxazoline monomer with pendant hydroxyl groups

Tert-butoxyacetic acid (2 g,15.1 mmol) was dissolved in 50mL of methylene chloride, EDCI (4.6 g,15.1 mmol), HOBT (3.3 g,15.1 mmol), N-2-isopropylethylamine (3.9 g,30.2 mmol) were sequentially added under ice bath, after stirring for 30 minutes, 2-chloroethylamine hydrochloride (2.64 g,22.6 mmol) was added, after stirring at room temperature for 8 hours, the reaction mixture was washed twice with 30mL of deionized water, 30mL of 5% aqueous citric acid, respectively, the organic phase was collected, dried over anhydrous magnesium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography to give intermediate (1.9 g, yield 65%).

Sodium hydroxide (3411 mg,8.5 mmol) was dissolved in 35mL ethanol in a dry single vial, then intermediate (1.5 g,7.7 mmol) was added and heated to reflux under nitrogen protection for 2h. The reaction solution was concentrated, and 30mL of methylene chloride was added for dissolution, washed twice with 15mL of deionized water, and finally purified by distillation under reduced pressure to give an O-tert-butyl protected hydroxy oxazoline monomer (850 mg, yield 70%). ¹H NMR(400MHz,CDCl₃ ) δ4.27 (t, j= 9.6,2H), 4.07 (s, 1H), 3.85 (t, j= 9.6,2H), 1.23 (s, 9H).

It should be understood that, as shown above, the raw material t-butoxyacetic acid is substituted for Cheng Shuding oxypropionic acid or t-butoxybutyric acid to prepare the hydroxyl oxazoline monomers with different carbon numbers in the side chains according to the described synthesis steps.

EXAMPLE 16 Synthesis of oxazoline homopolymer with pendant hydroxyl groups

The experimental procedure was identical to example 1, except that the N-Boc-2- (aminomethyl) oxazoline monomer was replaced with the hydroxy oxazoline monomer prepared in example 15, cooled to room temperature after the polymerization reaction was completed, cold petroleum ether (45 mL) was poured into the reaction mixture, the precipitated white floccules were collected by centrifugation, dried in air flow, redissolved in tetrahydrofuran (1.5 mL), and a large amount of cold petroleum ether was added for precipitation. This dissolution-precipitation procedure was repeated three times in total to give 52.2mg (yield 83%) of the oxazoline polymer with protected side chain hydroxyl groups.

The molecular weight mn=2980 and the molecular weight distribution PDI (Mw/Mn) =1.14 of the polymer obtained by MeOTf initiated polymerization were identified by Gel Permeation Chromatography (GPC).

Then adding 2mL of trifluoroacetic acid into the pumped polymer, slightly shaking at room temperature for about 2 hours, blowing off excessive trifluoroacetic acid, dissolving the obtained viscous liquid into 0.5mL of methanol, adding 45mL of frozen diethyl ether to separate out white precipitate, and repeating the dissolving-precipitating process for three times to obtain the oxazoline homopolymer with side chain hydroxyl groups. The deprotected polymer was again dissolved in 5mL of ultrapure water, and filtered and lyophilized for subsequent bioactivity testing.

EXAMPLE 17 application of 3- (triphenylmercapto) propyl bromide-initiated homo-polymerization of hydroxy oxazoline monomer as surface antifouling material

The polymer synthesis was carried out as in example 16, except that methyl triflate was replaced with 3- (triphenylmercapto) propyl bromide as initiator. After the polymerization reaction was completed, the mixture was cooled to room temperature, cold petroleum ether (45 mL) was poured into the reaction mixture, and the white floccule separated out was collected by centrifugation, dried in an air stream, redissolved in tetrahydrofuran (1.5 mL), and precipitated by adding a large amount of cold petroleum ether. The polymer is obtained by mixing two or more monomers synthesized by three dissolving-precipitating processes in a set proportion and copolymerizing. The polymer which is pumped out is added with 2mL of trifluoroacetic acid and 5% (v/v) of triethylsilane, after shaking slightly at room temperature overnight, excess trifluoroacetic acid is blown off, the obtained viscous liquid is dissolved in 0.5mL of methanol, 45mL of frozen diethyl ether is added to separate out white precipitate, and the dissolving-precipitating process is repeated three times, so that the oxazoline homopolymer with side chain hydroxyl groups and end group mercapto groups deprotected is obtained. The deprotected polymer was again dissolved in 5mL of ultrapure water and filtered and lyophilized for subsequent bioactivity testing.

The deprotected oxazoline polymer with side chain hydroxyl group was grafted on the surface of gold plate, and the surface antifouling test was carried out by culturing E.coli ESCHERICHIA COLI 25922 with LB liquid medium (Luria-Bertani Broth) in a shaking table at 37℃at 150rpm overnight. After the completion of the culture, 7.5mL of the bacterial liquid was taken out from the conical flask, centrifuged at 4000rpm for 5min to collect the bacteria, re-dispersed in PBS, re-centrifuged, and the bacterial liquid was collected after repeating the centrifugation of the PBS dispersion three times, and the number of colonies was quantified by reading absorbance at 600nm (OD ₆₀₀) with an ELISA reader. The bacterial liquid is diluted to 1X 10 ⁶ CFU/mL by PBS for standby. The prepared polymer soil resistant surface was placed in a 24 well plate and 100. Mu.L of 2mg/mL polymer solution was added to the gold plate surface with PBS as a control. After overnight, the mixture was alternately washed with deionized water and ethanol for 2 times and dried with nitrogen. Placing the cells in a new 24-well plate, adding 1mL of diluted bacterial liquid into each well, incubating for 1d, washing for multiple times by using PBS, and staining by LIVE/DEAD.

Observing the adhesion and growth state of bacteria on the surface of the polymer under an inverted microscope; the experimental results show that no significant bacteria are observed on the glass surface of the grafted oxazoline polymer, while a large number of bacteria are observed on the surface of the grafted polymer, so that the hydroxyl oxazoline homopolymer exhibits excellent surface antifouling material function.

Example 18 application of oxazoline homopolymer with side chain hydroxyl group as cell cryopreservation protective Material

Cell cryopreservation and resuscitation experiments different CPA formulations were prepared using cell culture medium containing 20% Fetal Bovine Serum (FBS) (pure medium as blank, medium+4% betaine, medium+4% betaine+0.2% oxazoline polymer, medium+10% dmso as positive control). After sterilization using a syringe filter apparatus (0.22 μm, titan), 0.5mL CPA solution with 1.0X10 ⁶ CFU/mL NIH3T3 cells was added to the freezer tube. Each sample was incubated at 37 ℃ for 1h and then transferred to a temperature-programmed box, which was transferred to an 80 ℃ refrigerator for freezing at a gradual freezing cooling rate of about-1 ℃/min. To test the activity of NIH3T3 cells after cryopreservation by the above method, after transferring the program cooling box to a-80 ℃ refrigerator for 12 hours, the freezing tube containing NIH3T3 cells was removed and rapidly thawed in a 37 ℃ water bath, NIH3T3 cells were collected by centrifugation at 1200rpm for 4min using a low speed centrifuge, the supernatant was finally removed, fresh medium was added to redisperse the cells in a petri dish, and the cells were transferred to a CO ₂ incubator and cultured in an environment of 37 ℃ and 5% CO ₂.

The oxazoline polymer with side chain hydroxyl groups prepared in example 16 is preferred, and the influence of the polymer on the activity of the cells after freezing and recovering is observed, and four cell culture mediums are prepared as pure culture medium, culture medium+10% DMSO, culture medium+4% betaine, and culture medium+4% betaine+0.2% oxazoline polymer. After the above described steps of cryopreservation and resuscitating of cells, the resuscitated NIH3T3 cells were placed in a CO ₂ incubator and cultured in an environment of 37 ℃ and 5% CO ₂ for 12 hours, and then the number of cell adhesion and cell adhesion morphology were observed using a random field under an inverted microscope. Experiment results show that NIH3T3 cells recovered after freezing and preserving pure culture medium and 4% betaine basically do not spread and grow on a culture dish, and the cell activity is negligible, which also shows that the pure culture medium and betaine cannot play a role in protecting during the freezing and preserving process of the cells; the recovered NIH3T3 cells are frozen in the traditional DMSO-added frozen stock solution, and a large amount of spreading can be observed on a culture dish; after the cells are frozen and recovered in the cell frozen solution added with the oxazoline polymer, a large number of NIH3T3 cells with good growth state can be observed on a culture dish, and the number of the cell spread obtained by a counting method is up to 95% of the NIH3T3 cells after the freezing and recovery in the traditional DMSO frozen solution. The experimental result shows that the surface hydroxyl oxazoline homopolymer shows the performance of the excellent cell cryopreservation protective material.

Further, it will be appreciated that various changes and modifications may be made by those skilled in the art after reading the above teachings, and such equivalents are intended to fall within the scope of the application as defined in the appended claims.

Claims

1. An oxazoline polymer comprising one or more of the repeating units of formula E ₁ and/or formula E ₂,

Wherein,

L ₁ and L' ₁ are each independently absent, -CHR ₁ -;

each R ₁ and each R' ₁ are each independently selected from the group consisting of substituted or unsubstituted: H. C1-C15 alkyl, C1-C15 alkylamino, C6-C15 aryl, 5-15 heteroaryl, 5-12 heterocyclyl, wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, -PhOH, C1-C15 alkyl, C1-C15 alkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl;

Ra and Rb are each independently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl, C1-C15 alkylamino, C1-C15 alkylhydroxy, C3-C12 cycloalkyl, 5-12 membered heterocyclyl, C6 aryl, 5-12 membered heteroaryl, wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, -PhOH, C1-C15 alkyl, C1-C15 alkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl;

Or in formula E2 Ra and Rb together with the N atom adjacent thereto form a substituted or unsubstituted 3-12 membered heterocyclyl, said 3-12 heterocyclyl containing at least 1N heteroatom and 0-2 heteroatoms selected from O and S; or Ra together with N and its adjacent C atoms forms a substituted or unsubstituted 3-12 membered heterocyclyl containing at least 1N heteroatom and 0-2 heteroatoms selected from O and S, wherein Rb is selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl, C1-C15 alkylamino, C1-C15 alkylhydroxy, C3-C12 cycloalkyl, 5-12 membered heterocyclyl, C6 aryl, 5-12 membered heteroaryl, wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, -PhOH, C1-C15 alkyl, C1-C15 alkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl;

m is 1,2,3 or 4;

m' is 1,2, 3 or 4;

q is an integer of 0, 1,2, 3, 4, 5;

q' is an integer of 0, 1, 2, 3; and m '+q'. Ltoreq.4;

wherein, for a substituted 3-12 membered heterocyclyl, the substitution refers to substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, glucosyl, C1-C15 alkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl.

2. The oxazoline polymer of claim 1 further comprising one or more of the B and/or C repeat units:

Wherein,

R ₂ is C1-C8 alkyl, C3-C8 cycloalkyl, C6-C12 aryl, C6-C12 arylalkyl;

Ra is independently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C15 alkyl;

l ₁、R₁, m and q are as defined in claim 1.

3. The oxazoline polymer of claim 1, wherein said formula E ₂ has a structure represented by formula E' ₂:

Wherein,

Wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, glucosyl, C1-C15 alkoxy, C3-C15 cycloalkyl, C4-C12 cycloalkenyl;

r '₁, m', rb are as defined in claim 1.

4. The oxazoline polymer of claim 1 or 2, wherein the oxazoline polymer is an oxazoline polymer having an amino group in a side chain, and has a structure as shown in formula II:

wherein n is an integer of 5 to 50000; 0% < x <100%; y is more than or equal to 0% and less than 100%; z is more than or equal to 0% and less than 100%; wherein, the calculation mode of x, y and z is the corresponding side chain repeating unit number divided by the total repeating unit number;

m, m', R ₁、R'₁, ra and Rb are as defined in claim 1; r ₂ is as defined in claim 2.

5. The oxazoline polymer of claim 1 or 2, wherein the oxazoline polymer is an oxazoline polymer having a hydroxyl group in a side chain, and has a structure as shown in formula I:

Wherein m, R ₁ and R _a are as defined in claim 1 and R ₂ is as defined in claim 2;

n is an integer of 5 to 50000; 0% < x <100%; y is more than or equal to 0% and less than 100%; z is more than or equal to 0% and less than 100%; wherein, the calculation mode of x, y and z is the corresponding number of side chain repeating units divided by the total number of repeating units.

6. The oxazoline polymer of claim 1 or 2, wherein the oxazoline polymer is an oxazoline polymer having an amino group and a hydroxyl group in a side chain, and has a structure as shown in formula III:

Wherein 0% < w <100%; x ₁ is more than or equal to 0% and less than 100%; y is more than or equal to 0% and less than 100%; z is more than or equal to 0% and less than 100%; wherein, the calculation mode of w, x ₁, y and z is the corresponding repeated unit number divided by the total repeated unit number;

m, m', R ₁、R'₁, ra and Rb are as defined in claim 1; r ₂ is as described in claim 2;

n is an integer of 5 to 50000.

7. The oxazoline polymer of claim 1 or 2, wherein the oxazoline polymer is an oxazoline polymer having an amino group and/or a hydroxyl group in a side chain, and is characterized in that it has a structure as shown in IV-IX:

Wherein n, x, y, z, m, R ₁ and R ₂ are as defined in claim 4;

Wherein n, x, y, z, m, m', R ₁、R'₁ and R ₂ are as defined in claim 4;

n, x, y, z, m, m', R ₁、R'₁、R₂ and Rb are as defined in claim 4, and ring a is as defined in claim 3;

wherein said substitution means substitution with one or more groups selected from the group consisting of: halogen, hydroxy, amino, guanidino, -CO-NH ₂, -COOH, ph-, 5-12 membered heteroaryl, 5-12 membered heterocyclyl;

n, x, y, z, m, R ₁ and R ₂ are as defined in claim 4;

Wherein,

M, m', n, x, y, z, w, x ₁、R₁、R'₁ and R ₂ are as defined in claim 4;

R ₆ is independently a substituted or unsubstituted C1-C15 alkyl-NRaRb, substituted or unsubstituted 5-12 membered heterocyclyl.

8. A process for the preparation of an oxazoline polymer having amino groups in the side chains, comprising the steps of:

Wherein R' ₁、m'、q'、L₁'、R₁、m、q、L₁, ra and Rb are as defined in claim 1; r ₂ is as described in claim 2;

The organic solvent is selected from the group consisting of: DMAc, DMF, or a combination thereof;

The initiator is selected from the group consisting of: meOTf, bzOTf, or a combination thereof.

9. An oxazoline monomer having a side chain comprising a hydroxyl group, characterized by having a structure represented by formula E' ₁:

wherein R ₁、m、q、L₁ and Ra are as defined in claim 1.

10. The oxazoline monomer having a hydroxyl-containing side chain of claim 9, wherein formula E' ₁ has a structure as shown in formula E "₁:

Wherein P ₂ is a hydroxyl protecting group selected from: TMS, TBS, TBDPS, t-Bu; m, R ₁ are as defined in claim 1.

11. A method of preparing an oxazoline monomer having a hydroxyl-containing side chain of claim 10, comprising the steps of:

Wherein m and P ₂、R₁ are defined as defined in claim 10;

x is a leaving group selected from the group consisting of: br, cl, I, OTs, OMs;

(2) Reacting compound 2 with a base in a second inert solvent to give a monomer of formula E "₁;

the base is selected from the group consisting of: sodium hydroxide, potassium hydroxide, triethylamine, N-diisopropylethylamine, or a combination thereof;

the second inert solvent is independently selected from the group consisting of: methanol, ethanol, acetonitrile, or a combination thereof.

12. The method of claim 11, further comprising the step of:

In the method, in the process of the invention,

R ₁、P₂, X, m are as defined in claim 11;

(1) Reacting compound 1 with H ₂N(CH₂)₂ X in a first inert solvent to give compound 2;

The first inert solvent is independently selected from the group consisting of: DCM, DMF, THF, DMAc, or a combination thereof.

13. A method for preparing an oxazoline polymer with a side chain containing hydroxyl, which is characterized by comprising the following steps:

Under the protection of inert solvent and inert gas, under the condition of initiator existence and heating, the oxazoline monomer represented by a formula E ' ₁ and optionally the oxazoline monomer represented by a formula B ' and/or a formula C ' are polymerized to obtain the oxazoline polymer with side chain hydroxyl,

Wherein R ₁、m、q、L₁, and Ra are as defined in claim 1; r ₂ is as described in claim 2;

the inert solvent is selected from the group consisting of: DMAc, DMF, meCN, phCN, or a combination thereof;

the initiator is selected from the group consisting of: meOTf, bzOTf, CH ₃(CH₂)₅Br、CH₃(CH₂)₅ OTs, or a combination thereof.

14. The preparation method of the oxazoline polymer with the side chain containing amino and hydroxyl comprises the following steps:

wherein R' ₁、m'、q'、L₁'、R₁、m、q、L₁, ra and Rb are as defined in claim 1;

15. Use of an oxazoline polymer having amino groups in the side chains, prepared by the preparation method of claim 8, for preparing materials for antibacterial, antifungal, antitumor, cell adhesion, drug synergy, drug delivery and self-assembled material fields.

16. Use of an oxazoline polymer having hydroxyl groups in the side chains, prepared by the preparation method of claim 13, for the preparation of an article or formulation for surface anti-fouling, cell cryoprotection.

17. Use of an oxazoline polymer having amino groups and hydroxyl groups in side chains, prepared by the preparation method of claim 14, for preparing a material having the functions of an oxazoline polymer having amino groups in side chains and hydroxyl groups in side chains, which is used for antibacterial, antifungal, antitumor, cell adhesion, drug synergy, drug delivery, self-assembled material, surface antifouling, cell cryoprotection.