Use of polymers to promote endothelialization of interventional / implant materials

Abstract

The present invention provides the use of a polymer in promoting endothelialization of an interventional / implant material. In particular, there is provided the use of a polymer having a segment selected from Formula I or Formula II, or a pharmaceutically acceptable salt thereof, in the preparation of an interventional / implant material for promoting endothelialization. The polymer of the present invention is capable of selectively promoting the adhesion and proliferation of endothelial cells, and is very suitable for the preparation of an interventional / implant material for promoting endothelialization.

Description

Technical Field

[0001] This invention relates to the medical field, and more specifically to the application of the polymers of this invention in promoting endothelialization of interventional / implantable materials. Background Technology

[0002] Cardiovascular diseases (CVDs) are illnesses caused by abnormalities and disorders of the heart or cardiovascular system, accounting for three-quarters of global deaths. Most CVDs are caused by atherosclerotic plaques. When these plaques rupture, the released lipids come into contact with the blood, leading to the formation of thrombi or blood clots that can block arteries. This can trigger acute conditions such as myocardial infarction.

[0003] Currently, the main treatments for atherosclerosis are drug therapy and surgical treatment. Surgical treatment includes stent placement and vascular transplantation. Stent placement involves implanting a stent to dilate the blood vessel, restoring blood supply to the artery. Vascular transplantation involves using autologous or artificial blood vessels to create a new vascular bypass, thereby improving blood supply. Both treatments require the integration and remodeling of the interventional / implanted materials with the vascular tissue, establishing a healthy endothelial layer with corresponding biological functions at the blood-contact interface. This effectively prevents thrombosis and restenosis, ensuring long-term vascular patency. Therefore, promoting rapid endothelialization of the surface of interventional / implanted materials is a crucial issue that urgently needs to be addressed.

[0004] Immobilizing bioactive molecules, such as extracellular matrix (ECM), peptides (REDV, YIGSR, CAG, etc.), or growth factors, through surface physical adsorption or chemical grafting can effectively promote endothelial cell adhesion and proliferation, accelerating the endothelialization process. However, the widespread application of these natural biomolecules is limited by their inherent structural instability (easily hydrolyzed by proteases), high cost, and inability to be prepared in large quantities.

[0005] Therefore, there is an urgent need in this field to develop compounds and interventional / implantable materials that are highly stable and can selectively promote the adhesion and proliferation of endothelial cells. Summary of the Invention

[0006] The object of this invention is to provide the use of a polymer or a pharmaceutically acceptable salt thereof in promoting endothelialization of interventional / implantable materials.

[0007] In a first aspect, the present invention provides the use of a polymer, or a pharmaceutically acceptable salt thereof, in the preparation of interventional / implantable materials for promoting endothelialization, wherein the polymer comprises segments having a selection from Formula I or Formula II:

[0008]

[0009]

[0010] In Equation I,

[0011] n is a positive integer from 1 to 100;

[0012] r1 and r2 are each independently 1 or 2;

[0013] Any one of R1 can be set to -L1R a L1 is selected from the group consisting of: bond, substituted or unsubstituted C1-C8 alkylene group, substituted or unsubstituted C2-C8 alkenyl group, and substituted or unsubstituted C2-C8 ynylene group; R a Selected from the following group: -NR b R b guanidine, biguanide, -N(R) b )3 + And each R b Independently selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 ynyl, substituted or unsubstituted C1-C6 alkyl-C3-C6 cycloalkyl; or two R atoms on the same N atom. b It forms a 4-8 membered heterocyclic group containing one nitrogen atom with the attached nitrogen atom;

[0014] Each R1 is not -L1R a The remaining groups are each independently selected from the group consisting of: H, halogen, -OH, substituted or unsubstituted C1-C15 alkyl, substituted or unsubstituted C2-C15 alkenyl, substituted or unsubstituted C2-C15 alkynyl, or substituted or unsubstituted C1-C6 alkyl-Rc, or substituted or unsubstituted C1-C6 alkyl-COO-Rc, and each Rc is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, S, substituted or unsubstituted 5-10 heteroaryl comprising 1-3 heteroatoms selected from O, N, S; or two R2s forming a substituted or unsubstituted C3-C12 cycloalkyl with the attached carbon atom;

[0015] Each R2 is independently selected from the group consisting of: H, halogen, -OH, substituted or unsubstituted C1-C15 alkyl, substituted or unsubstituted C2-C15 alkenyl, substituted or unsubstituted C2-C15 alkynyl, or substituted or unsubstituted C1-C6 alkyl-Rc, or substituted or unsubstituted C1-C6 alkyl-COO-Rc, and each Rc is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, S, or substituted or unsubstituted 5-10 heteroaryl comprising 1-3 heteroatoms selected from O, N, S; or two R2s forming a substituted or unsubstituted C3-C12 cycloalkyl with the attached carbon atom.

[0016] In Equation II, n is a positive integer from 1 to 100;

[0017] R3 is -L1R a L1 is selected from the group consisting of: bond, substituted or unsubstituted C1-C8 alkylene group, substituted or unsubstituted C2-C8 alkenyl group, and substituted or unsubstituted C2-C8 ynylene group; R a Selected from the following group: -NR b R b guanidine, biguanide, -N(R) b )3 + And each R b Independently selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 ynyl, substituted or unsubstituted C1-C6 alkyl-C3-C6 cycloalkyl; or two R atoms on the same N atom. b It forms a 4-8 membered heterocyclic group containing one nitrogen atom with the attached nitrogen atom;

[0018] R4 is selected from the following group: H, halogen, -OH, substituted or unsubstituted C1-C15 alkyl, substituted or unsubstituted C2-C15 alkenyl, substituted or unsubstituted C2-C15 ynyl, substituted or unsubstituted C3-C12 cycloalkyl, substituted or unsubstituted 3-12 heterocyclic alkyl comprising 1-3 heteroatoms selected from O, N, and S, substituted or unsubstituted benzyl, substituted or unsubstituted phenyl, substituted or unsubstituted comprising 1-3 heteroatoms selected from O, N, and S. The heteroatom is a 5-10 membered heteroaryl, or a substituted or unsubstituted C1-C6 alkyl-Rc, or a substituted or unsubstituted C1-C6 alkyl-COO-Rc, wherein each Rc is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 heterocycloalkyl including 1-3 heteroatoms selected from O, N, S, or substituted or unsubstituted 5-10 membered heteroaryl including 1-3 heteroatoms selected from O, N, S;

[0019] Unless otherwise specified, in each formula, “substitution” means that one or more hydrogen atoms on each group are independently substituted by a group selected from the group consisting of: deuterium, halogen, -COOH, -OH, -NH2, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkoxy, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated benzyl, unsubstituted or halogenated 3-6 heterocyclic alkyl comprising 1-3 heteroatoms selected from O, N, and S, unsubstituted or halogenated C6-C10 aryl, and unsubstituted or halogenated 5-6 heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

[0020] In another preferred embodiment, in each formula, -L1R a Independently -C2-C6 alkyl-R a Preferably, -C3-C6 alkyl-R a .

[0021] In another preferred embodiment, R a Selected from the following group: -NH2, guanidino Biguanide, -NH3 + -N(CH3)3 + and -N(C2H5)3 + .

[0022] In another preferred embodiment, r1 and r2 are 1, or r1 and r2 are 2, or r1 is 1 and r2 is 2, or r1 is 2 and r2 is 1.

[0023] In another preferred embodiment, any one of R1 is chosen as -L1R. a The remaining R1s are each independently selected from the following groups: H, C1-C6 alkyl, C1-C6 alkyl-C3-C6 cycloalkyl.

[0024] In another preferred embodiment, at least one R2 group is selected from the group consisting of substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 ynyl, substituted or unsubstituted -C1-C6 alkyl-Rc; and Rc is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, benzyl, phenyl, C3-C8 cycloalkyl; or two R2 groups form a substituted or unsubstituted C3-C12 cycloalkyl group with the attached carbon atom.

[0025] In another preferred embodiment, each R4 is selected from the group consisting of: substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted -C1-C6 alkyl-Rc; and Rc is selected from the group consisting of: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, benzyl, phenyl, C3-C8 cycloalkyl.

[0026] In another preferred embodiment, x is independently 10-100, more preferably 10-90, such as 20, 30, 40, 50, 60, 70 or 80.

[0027] In another preferred embodiment, y is independently 5-90, more preferably 10-90, such as 20, 30, 40, 50, 60, 70 or 80.

[0028] In another preferred embodiment, n is independently a positive integer from 5 to 50, such as 10, 15, 20, 25, 30, 35, or 40.

[0029] In another preferred embodiment, the polymer comprises segments selected from the group consisting of:

[0030]

[0031]

[0032]

[0033] In each expression, x is independently between 5 and 100; and x + y = 100; n is a positive integer between 1 and 100.

[0034] In another preferred embodiment, the polymer is the polymer used in the examples.

[0035] In another preferred embodiment, the polymer is a segment having a composition of NM:CH 40:60.

[0036] In another preferred embodiment, the polymer is selected from the group consisting of:

[0037]

[0038] In each expression, x is independently between 5 and 100; and x + y = 100; n is a positive integer between 1 and 100.

[0039] In another preferred embodiment, the average molecular weight Mn of the polymer is 1000-8000, more preferably 2000-6000, and even more preferably 3000-5200, such as about 3500, 4000, 4500 or 5000.

[0040] In another preferred embodiment, the molecular weight distribution (PDI) of the polymer is independently 1-1.4, more preferably 1.1-1.3, and even more preferably 1.1-1.25.

[0041] In another preferred embodiment, the polymer is a random copolymer or a block copolymer.

[0042] In another preferred embodiment, the polymer also has two end groups.

[0043] In another preferred embodiment, one end of the compound includes a terminal group having a reactive group selected from the group consisting of: -SH, -NH2, -COOH, -Br, -Cl, -OH, epoxy, alkenyl, alkynyl, -COCl, azide, maleimide, or o-dithiopyridyl (OPSS). For example, -C1-C6 alkyl-SH, -NH-C1-C6 alkyl-SH, -CO-C1-C6 alkyl-SH.

[0044] In another preferred embodiment, the other end group of the polymer is selected from the group consisting of H, monomer residues, and initiator residues. Those skilled in the art will understand that the monomer residues, initiator residues, etc., on the end group are related to the type of monomer and the type of initiator, but will not significantly affect the activity of the polymer chain segment of the present invention.

[0045] In another preferred embodiment, the initiator is selected from the group consisting of: LiHMDS, NaHMDS, KHMDS, triphenylmethylmercaptoethylamine, p-tert-butylbenzylamine, and triphenylmercapto-3-bromopropyl.

[0046] In another preferred embodiment, the monomer residues and initiator residues may be capped by a capping agent, preferably selected from the group consisting of triphenylmethylmercaptoethylamine and p-tert-butylbenzylamine.

[0047] In another preferred embodiment, the pharmaceutically acceptable salt is selected from the group consisting of: hydrochloride, bromate, trifluoroacetate, phosphate, lithium, sodium, and potassium salts.

[0048] In another preferred embodiment, the substrate surface before modification of the interventional / implantable material is selected from the group consisting of: inorganic non-metallic biomaterials (such as bioceramics, bioglass, graphene, bone cement, and medical carbon materials), biometallic materials (such as stainless steel, cobalt-based and titanium-based alloys, shape memory alloys, precious metals such as silver, platinum, tantalum, niobium, zirconium, palladium, and platinum), natural polymer materials (such as hyaluronic acid, chitosan, alginate, cellulose, collagen, and gelatin), synthetic polymer materials (polyetheretherketone, polycaprolactone, polylactic acid, polycarbonate, polyurethane, polyester, polyanhydride, polydimethylsiloxane, polymethyl methacrylate, polyphosphazene, polyamide, polyethylene, polypropylene, polytetrafluoroethylene, polyethylene terephthalate, and resins), or combinations thereof (such as composite materials and assembly materials thereof).

[0049] In another preferred embodiment, the interventional / implantable material is selected from the group consisting of: cardiovascular stents and artificial blood vessels.

[0050] A second aspect of the present invention provides an interventional or implantable material, characterized in that the substrate surface of the interventional or implantable material is coated or covalently modified with segments selected from Formula I or Formula II, or a polymer containing thereto.

[0051] The use of the interventional or implantable material, as described in the third aspect of the present invention, is for promoting endothelialization after intervention or implantation.

[0052] A fourth aspect of the present invention provides a method for enabling interventional or implantable materials to promote endothelialization, characterized by comprising the step of: coating or covalently modifying at least a portion of the surface of the material substrate with a segment selected from Formula I or Formula II, or a polymer containing thereto.

[0053] In another preferred embodiment, the method is in vitro non-diagnostic and non-therapeutic.

[0054] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0055] Figure 1 For surface characterization of materials;

[0056] Figure 2-1 , 2-2 Figures 2 and 3 show the full adhesion screening of HUVECs on the polymer-modified surface of the present invention;

[0057] Figure 3-1 , 3-2 Figures 3-3 show the full adhesion screening of the polymer-modified surface HUASMC of the present invention;

[0058] Figure 4-1 , 4-2 Figures 4-3 show the HUVEC proliferation on the polymer-modified surface of the present invention;

[0059] Figure 5-1 , 5-2 Figures 5-3 show the proliferation of HUASMC on the polymer-modified surface of the present invention;

[0060] Figure 6 This invention illustrates the co-cultivation of HUVEC and HUASMC on polymer-modified surfaces.

[0061] Figure 7 This illustrates cell migration after surface grafting modification of the polymer according to the present invention;

[0062] Figure 8The illustration shows the endothelialization of the stent surface during an in vivo rat experiment using the polymer-modified vascular stent mimicry of the present invention. Detailed Implementation

[0063] Through extensive and in-depth research, including numerous screenings and tests, the inventors have provided the use of the polymer of this invention in promoting endothelialization of interventional / implantable materials. The inventors unexpectedly discovered that modifying the surface of materials with the polymer of this invention can selectively promote endothelial cell adhesion and proliferation, thereby promoting endothelialization of interventional / implantable materials and preventing endothelial cell hyperplasia, thus ensuring the safe, durable, and stable placement of interventional / implantable materials in the body. This invention was completed based on this discovery.

[0064] the term

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0066] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0067] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0068] As used herein, the term “room temperature” or “normal temperature” refers to a temperature of 4–40°C, preferably 25 ± 5°C.

[0069] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0070] When a substituent is described using a conventional chemical formula written from left to right, it also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0071] Throughout this specification, the terms “optionally substituted” or “may be substituted” indicate that the group may or may not be further substituted or fused with one or more non-hydrogen substituents (to form a polycyclic system). Substituents for suitable chemically appropriate specific functional groups will be apparent to those skilled in the art.

[0072] As used in this text, the term "alkyl" refers to a straight-chain or branched alkyl group containing a number of carbon atoms, wherein "C1-C..."15 "Alkyl" refers to a straight-chain or branched alkyl group having 1 to 15 carbon atoms, including alkyl groups with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 carbon atoms. Alkyl groups are preferably, for example, C1-C2, C1-C3, C1-C4, C1-C5, C1-C6, C1-C7, C1-C8, C1-C9, or C1-C6. 10 , 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" compounds include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, and isobutyl. Amyl, isopentyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, etc. In this invention, alkyl also includes substituted alkyl groups. "Substituted alkyl" means that one or more positions in an alkyl group are substituted, particularly 1-4 substituents, which can be substituted at any position.

[0073] As used in the text, the term "C1-C" 15 "Alkoxy" refers to a straight-chain or branched alkoxy group having 1-15 carbon atoms, with C1-C2 atoms. 15 Alkyl-O- structure, C1-C 15 Alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, and hexoxy, with ethoxy being preferred. C1-C 15 Alkoxy groups also include substituted C1-C groups. 15 Alkyl group.

[0074] As used in the text, the term "C1-C" 15 "alkyl hydroxyl" refers to -C1-C 15 alkylene -OH, -C1-C 15 Alkylenes have the definition described above, C1-C 15 Alkyl hydroxyl groups include, but are not limited to, -CH2OH and -CH2CH2OH. (C1-C) 15 Alkyl hydroxyl groups also include substituted C1-C hydroxyl groups. 15 Alkyl hydroxyl.

[0075] As used in the text, the term "C1-C" 15 "alkylsulfonyl" refers to C1-C 15 Alkyl S(=O)2-.

[0076] As used in the text, the term "C1-C" 15 Alkyl-C6-C 15 "Aryl" refers to -C1-C15 Alkyl-C6-C 15 Aryl groups, such as -CH2CH2CH2Ph ​​and -Bn.

[0077] As used in the text, the term "C1-C" 15 "alkyl ester group" refers to C1-C 15 Alkyl C(=O)-O- or -C(=O)-O-C1-C 15 alkyl.

[0078] As used in this text, the term "thio-C1-C15 alkyl ester" refers to C1-C 15 Alkyl C(=S)-O- or -C(=S)-O-C1-C 15 alkyl.

[0079] As used in this text, the term "guanidinium" refers to NHC(=NH)NH-, and the term "biguanide" refers to -NH-C(CH)-NH-C(CH)-NH2.

[0080] As used in this text, the term "alkenyl" refers to a straight-chain or branched hydrocarbon having at least two carbon atoms and at least one double bond. Alkenyl groups can include any number of carbon atoms, where "C2-C15 alkenyl" refers to a straight-chain or branched hydrocarbon having 2-15 carbon atoms and at least one double bond, such as C2, C15, etc. 2- C3, C 2- C4, C 2- C5, C 2- C6, C 2- C7, C 2- C8, C 2- C9, C 2- C 10 C3, C 3- C4, C 3- C5, C 3- C6, C4, C 4- C5, C 4- C6, C5, C 5- C6 and C6. The alkenyl group 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-hextrienyl. Like the alkyl groups described above, the alkenyl group may be substituted or unsubstituted.

[0081] As used in this text, the term "alkynyl" refers to a straight-chain or branched hydrocarbon having at least two carbon atoms and at least one triple bond. Alynyl groups can include any number of carbon atoms; "C2-C15 alkynyl" refers to a straight-chain or branched hydrocarbon having 2-15 carbon atoms and at least one triple bond, such as C2, C15, ... 2- C3, C 2- C4, C 2- C5, C 2- C6, C 2- C7, C 2- C8, C 2- C9, C 2- C 10 C3, C 3- C4, C 3- C5, C 3- C6, C4, C 4- C5, C 4- C6, C5, C 5- C6 and C6. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butyrynyl, 1-pentynyl, 2-pentynyl, isopentenynyl, 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-hextriynyl. Like the alkyl groups described above, alkynyl groups can be substituted or unsubstituted.

[0082] The term "aryl" refers to an aromatic cyclic hydrocarbon group. "C6-C12 aryl" refers to an aromatic cyclic hydrocarbon group containing 6, 7, 8, 9, 10, 11, or 12 ring carbon atoms, having 1-3 rings, especially monocyclic and bicyclic groups, such as phenyl, biphenyl, or naphthyl. Any aryl group containing two or more aromatic rings (bicyclic, etc.) can have its aromatic rings linked by single bonds (e.g., biphenyl) or fused (e.g., naphthalene, anthracene, etc.). "Substituted aryl" refers to an aryl group where one or more positions are substituted, especially 1-3 substituents, which can be substituted at any position.

[0083] As used in this text, the term "heteroaryl" refers to a heteroaryl system containing 1-3 atoms selected from N, O, and S atoms, wherein "5-12-membered heteroaryl" refers to a 5-12-membered heteroaryl system containing 1-3 atoms selected from N, O, and S atoms. The heteroaryl group is preferably a 5- to 10-membered ring, more preferably a 5- or 6-membered ring, and includes, but is not limited to, pyrroleyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl, and tetrazolyl. "Heteroaryl" can be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, deuteralkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxyl, mercapto, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkylthio, oxo (-oxo), carboxyl and carboxylic ester groups.

[0084] As used in this text, the term "cycloalkyl" refers to a fully saturated cyclic hydrocarbon group having several carbon atoms, wherein "C3-C..." 12 "Cycloalkyl" refers to a fully saturated cyclic hydrocarbon group having 3-12 carbon atoms, preferably C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, C3-C9, or C3-C6. 10 "Replacing C3-C" 12 "Cycloalkyl" refers to a cycloalkyl group in which one or more positions are substituted, particularly 1-4 substituents, which can be substituted at any position, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. In this invention, "cycloalkyl" is intended to include "substituted cycloalkyl".

[0085] As used in this text, the term "cycloalkenyl" refers to an unsaturated cyclic hydrocarbon group with 1-3 double bonds having several carbon atoms, wherein "C4-C 12 "Cycloalkenyl" refers to an unsaturated cyclic hydrocarbon group with 1-3 double bonds and 4-12 carbon atoms, preferably C6-C. 10 Cycloalkenyl, C4-C6 cycloalkenyl, including but not limited to cyclobutenyl, cyclopentenyl, and cyclohexenyl.

[0086] As used in this text, the term "heterocyclic group" refers to a fully saturated or partially unsaturated cyclic group having several (greater than or equal to 3) ring atoms and 1-3 heteroatoms. Specifically, "5-12 membered heterocyclic group" refers to a fully saturated or partially unsaturated cyclic group having 5-12 ring atoms and 1-3 heteroatoms (including, but not limited to, 3-7 membered monocyclic, 6-11 membered bicyclic, or 8-12 membered tricyclic systems). Nitrogen or sulfur atoms may be oxidized, and nitrogen atoms may be quaternized. Heterocyclic groups can be attached to any heteroatom or carbon atom residue in a ring or cyclic molecule. Typical monocyclic heterocycles include, but are not limited to, nitrogen-containing heterocyclic butyl, pyrrolyl, oxoheterocyclic butyl, pyrazolinyl, imidazolinyl, imidazolinyl, oxazolinyl, isoxazolinyl, thiazolinyl, isothiazolinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopiperylyl, hexahydroacoxaneyl, 4-piperidinoneyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxaneyl, and tetrahydro-1,1-dioxothiophene, etc. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups; wherein the spirocyclic, fused-ring, and bridged-ring heterocyclic groups involved are optionally connected to other groups via single bonds, or further cyclically linked to other cycloalkyl, heterocyclic, aryl, and heteroaryl groups via any two or more atoms on the ring; the heterocyclic group can be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, deuteralkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxyl, mercapto, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkylthio, oxo, carboxyl, and carboxylic acid ester groups. Heterocyclic groups include, but are not limited to: tetrahydropyrrole, tetrahydrofuranyl, piperidinyl, piperazine, etc.

[0087] When a substituent is a non-terminal substituent or when a related group loses one hydrogen atom, it becomes a subunit of the corresponding group, usually a divalent group. The above definition of a group also applies to its divalent, trivalent, and other polyvalent groups. For example, an alkyl group losing one hydrogen atom becomes an alkylene group (e.g., methylene, ethylene, propylene, isopropylene). ), butylide (such as) ), pentylene (e.g.) ), hexyl (such as) ), subheptagen (such as ) etc.), cycloalkyl corresponding to cycloalkylene (e.g.: (etc.), heterocyclic groups corresponding to subheterocyclic groups (such as: ), cycloalkyl corresponding heterocyclic groups (e.g.: (etc.), alkoxy groups corresponding to alkeneoxy groups (-CH2O-, -CH2CH2-O-CH2-, -CH2OCH2CH2CH2-, etc.).

[0088] As described in this article, the term "multiple" refers to two or more, such as 2, 3, 4, 5, or 6.

[0089] As described herein, the compounds / polymers of this invention can be expanded with any number of substituents or functional groups. Generally, the term "substitution," whether preceding or following the term "optional," in this invention includes the general formula for substituents, meaning the replacement of a hydrogen radical with a substituent of a specified structure. When multiple positions in a particular structure are replaced by multiple specific substituents, each position of the substituent can be the same or different. The term "substitution" as used herein includes all permissible substitutions in organic compounds. In a broad sense, permissible substituents include acyclic, cyclic, branched-unbranched, carbocyclic, and heterocyclic, aromatic and non-aromatic organic compounds. In this invention, heteroatomic nitrogen may be supplemented with hydrogen substituents or any permissible organic compound described above to complete its valence state. Furthermore, this invention is not intended to limit permissible substituted organic compounds in any way.

[0090] As used herein, the term “substituted” means that any one or more hydrogen atoms on a specified atom are substituted by a substituent selected from the specified substituents, provided that the substitution does not exceed the normal valence of the specified atom, and the resulting compound is stable, i.e., can be isolated, characterized, and tested for bioactivity.

[0091] Unless otherwise specified, the substitution refers to the independent substitution of one or more H atoms on each group by a group selected from the group consisting of: deuterium, halogen, -SH, -OH, -NH2, benzyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C6 haloalkenyl, C2-C6 haloalkynyl, C3-C8 cycloalkyl, and C6-C10 aryl.

[0092] Unless otherwise stated, it is assumed that any heteroatom in a suboptimal valence state has enough hydrogen atoms to compensate for its valence state.

[0093] As used in this text, the terms "statistic coplymer" or "random" refer to polymers formed by the random linkage of two or more monomers polymerized simultaneously.

[0094] As used in this text, the term "block polymer" refers to a polymer formed by the sequential polymerization of two or more monomers, which consists of different chain segments connected together, wherein at least one monomer has an amino or hydroxyl side chain.

[0095] Unless otherwise specified, all compounds / polymers mentioned in this invention are intended to include all possible optical isomers, such as compounds with a single chirality, or mixtures of various chiral compounds (i.e., racemates). In all compounds of this invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R and S configurations.

[0096] As used herein, the term "pharmaceutically acceptable salt" means that the compounds of the present invention can be used in the form of salts derived from pharmaceutically or physiologically acceptable acids or bases. These salts include (but are not limited to) salts formed with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, succinic acid, oxalic acid, fumaric acid, maleic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or hydroxyethanesulfonic acid. Other salts include salts formed with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium), and salts in the form of esters, carbamates, or other conventional "prodrugs."

[0097] Active ingredients

[0098] In this invention, the active ingredient is a polymer or a pharmaceutically acceptable salt thereof, wherein the polymer comprises segments selected from Formula I or Formula II:

[0099]

[0100] In Equation I, r1, r2, R1, R2, x, y, and n are defined as above;

[0101]

[0102] In Equation II, R3, R4, x, y, and n are defined as above.

[0103] Interventional / Implantable Materials and Their Uses

[0104] In this invention, there are no particular requirements regarding the type of interventional implant material; it can be any interventional / implant material of any shape and material commonly used in the art. For example, the substrate surface before modification of the interventional / implant material can be selected from the following group (but is not limited to): inorganic non-metallic biomaterials (such as bioceramics, bioglass, graphene, bone cement, and medical carbon materials), biometallic materials (such as stainless steel, cobalt-based and titanium-based alloys, shape memory alloys, precious metals such as silver, platinum, tantalum, niobium, zirconium, palladium, and platinum), natural polymer materials (such as hyaluronic acid, chitosan, alginate, cellulose, collagen, and gelatin), synthetic polymer materials (polyetheretherketone, polycaprolactone, polylactic acid, polycarbonate, polyurethane, polyester, polyanhydride, polydimethylsiloxane, polymethyl methacrylate, polyphosphazene, polyamide, polyethylene, polypropylene, polytetrafluoroethylene, polyethylene terephthalate, and resin), or combinations thereof (such as composite materials and assembly materials thereof).

[0105] Typically, the interventional / implanted materials are (but are not limited to): cardiovascular stents, artificial blood vessels, bone plates, bone screws, bone pins, bone rods, spinal fixation devices, catheters, etc.

[0106] Promoting in-situ endothelialization of cardiovascular implant materials is an effective means to address adverse reactions such as thrombosis, inflammation, and restenosis after traditional implantation. The polymer modification of the material surface in this invention selectively promotes endothelial cell adhesion and proliferation, facilitating endothelialization of interventional / implantable materials while preventing endothelial cell hyperplasia, thereby ensuring the safe, durable, and stable placement of interventional / implantable materials within the body.

[0107] As used in this invention, "easy to endothelialize" or "for promoting endothelialization" means that the modified interventional / implant material improves the rate or extent of endothelialization by at least 10%, at least 20%, at least 50%, or even 100% compared to interventional / implant material without modification of the active ingredient of this invention.

[0108] Modification methods

[0109] In this invention, the active ingredient can be coated or covalently modified onto the substrate surface using methods commonly used in the art.

[0110] Typically, covalent modification may include the following steps: (i) pre-activating at least a portion of the surface of the substrate to form an anchoring layer; (ii) reacting the polymer of the present invention, having end groups capable of reacting with the anchoring layer, with the anchoring layer to modify the chain segments onto the material surface. After modification, the steps may further include: sealing reaction sites and / or washing and drying.

[0111] In another preferred embodiment, the pre-activation method may be selected from the following group:

[0112] (a) A plasma irradiation activation method, comprising the steps of: placing a substrate in a low-temperature plasma and activating it to obtain a pre-activated surface containing oxygen free radicals, hydroxyl groups, carboxyl groups, epoxy groups and / or aldehyde groups; placing the pre-activated surface obtained by the above method (a) in a bromoform solution to obtain a pre-activated surface with surface bromination.

[0113] (b) A pre-activated surface with surface amination is obtained by irradiating the substrate surface with ultraviolet ozone and then placing it in a solution of 3-aminopropyltriethoxysilane (APTES); or

[0114] (c) The substrate is placed in an alkaline solution of dopamine to obtain a dopamine pre-activated surface. Specifically, the dopamine pre-activation conditions may be: the solvent of the alkaline solution is tris(hydroxymethyl)aminomethane hydrochloride; the pH of the alkaline solution is 8.5-10, such as 9 or 9.5.

[0115] In another preferred embodiment, the pre-activation temperature of each pre-activation method is independently 0-120°C, such as 10-100°C, 20-60°C, or 30-40°C. In another preferred embodiment, the pre-activation time of each pre-activation method is independently 5 min-48 h, such as 10 min, 30 min, 1 h, 2 h, 6 h, 12 h, or 24 h.

[0116] Typically, the polymer has end groups selected from the group consisting of reactive groups, or the segments can be modified to have reactive groups selected from the group consisting of: -SH, -NH2, -COOH, -Br, -Cl, -OH, epoxy, alkenyl, alkynyl, -COCl, azido, maleimide, and o-dithiopyridyl (OPSS). The reactive groups can be linked to the polymer segments via linking groups such as C1-C6 alkylene, -CO-C1-C6 alkylene, and -NH-C1-C6 alkylene. The reactive groups are used to react with reactive groups on the surface of a pre-activated substrate, thereby attaching the polymer segments of the present invention to the substrate surface. The selection of the reactive groups of the compound based on the pre-activation method, or the selection of the pre-activation method based on the reactive groups of the compound, is familiar to those skilled in the art. For example, it can be selected from nucleophilic substitution reactions based on thiol-halogen atoms, addition reactions of thiol-epoxy groups, addition reactions of thiol-alkenyl / alkynyl or azido-alkynyl groups, etc.

[0117] In another preferred embodiment, in step (ii), the reaction is carried out by immersing the pre-activated surface in a solution of the compound or its salt, preferably, the concentration of the compound or its salt in the solution is 0.1-10 mg / mL, more preferably 0.5-5 mg / mL, such as 1 mg / mL, 2 mg / mL, 3 mg / mL, or 4 mg / mL. The reaction of the polymer of the present invention on the pre-activated surface will automatically reach saturation or equilibrium. For example, the average grafting density of the compound on at least a portion of the surface of the material is ≥0.1 chain / nm. 2 Preferably, ≥0.2 chain / nm 2 or ≥0.23chain / nm 2 The average grafting density of the compound on at least a portion of the surface of the material is 0.1-2 chains / nm. 2 ,

[0118] 0.15-1 chain / nm 2 0.2-0.5 chain / nm 2 Or, for example, 0.25 chains / nm 2 0.3chain / nm 2 0.35chain / nm 2 or 0.4chain / nm 2.

[0119] In another preferred embodiment, the surface contact angle of at least a portion of the substrate surface is reduced by 20-100° compared to the untreated substrate surface, more preferably by 30-80°, 40°, 45°, 50°, 55°, 60°, 65° or 75°, and preferably, the surface contact angle of the modified material is 45-65°, such as 50°, 55° or 60°.

[0120] The main advantages of this invention include:

[0121] 1. The chain segments or polymers of the present invention are a class of known substances that are stable, safe and non-toxic, and have certain bactericidal and other biological activities, making them very suitable for use in vivo. Compared with natural molecules, they can tolerate various covalent modification conditions commonly used in the field, making them suitable for industrial production and application.

[0122] 2. The segments or polymers of the present invention have excellent endothelialization-promoting effects on materials, and can selectively promote the adhesion and proliferation of endothelial cells, thereby enabling the interventional / implantable materials to be safely, permanently, and stably placed in the patient's body.

[0123] 3. The segments or polymers of the present invention can prevent the proliferation of endothelial cells, and while endothelializing the interventional / implantable material, they can maintain the patency of blood vessels to the maximum extent, thereby achieving better therapeutic effects.

[0124] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

[0125] Synthesis of α-peptide polymers with terminal thiol groups

[0126] Example 1

[0127] Random α-amino acid copolymers were prepared from N-ε-tert-butoxycarbonyl-DL-lysine-N-carboxyl ring anhydride and DL-norleucine-N-carboxyl ring anhydride. Rac Lys x : Rac Nle y )

[0128]

[0129] Accurately weigh 33.4 mg (0.2 mmol) of hexamethyldisilamide lithium salt and prepare a 0.1 M solution with 2 mL of tetrahydrofuran for later use. Weigh N-ε-tert-butoxycarbonyl-DL-lysine-N-carboxycyclic anhydride and DL-norleucine-N-carboxycyclic anhydride, using tetrahydrofuran as the solvent. Mix 1.8 mL of N-ε-tert-butoxycarbonyl-DL-lysine-N-carboxycyclic anhydride (0.2 M) and 0.2 mL of DL-norleucine-N-carboxycyclic anhydride (0.2 M) and stir with a magnetic stir bar (taking a monomer ratio of x:y = 9:1 as an example). Add 0.8 mL of the 0.1 M hexamethyldisilamide lithium salt solution to the stirred reaction flask. Stir the mixture in a glove box at room temperature for 5 min. After the polymerization reaction is complete, end-cap with triphenylmercaptoethylamine overnight. Cold petroleum ether (45 mL) was added to the above reaction mixture. The precipitated white flocculent precipitate was collected by centrifugation, dried in an air stream, and redissolved in tetrahydrofuran (1.5 mL). A large amount of cold petroleum ether was then added to precipitate the precipitate. This dissolution-precipitation process was repeated three times to obtain the purified copolymer. The molecular weight of the polymer, Mn = 7500 g / mol, and the molecular weight distribution Mw / Mn = 1.19 were identified by gel permeation chromatography (GPC).

[0130] The dried polymer was added to 2 mL of trifluoroacetic acid and 5% (v / v) triethylsilane. After gently shaking overnight at room temperature, excess trifluoroacetic acid was blown off, and the resulting viscous liquid was dissolved in 0.5 mL of methanol. Then, 45 mL of methoxymethyl tert-butyl ether was added to precipitate a white precipitate. The dissolution-precipitation process was repeated three times to obtain a random polymer with deprotected side-chain amino groups (yield 85%). The deprotected polymer was dissolved again in 5 mL of ultrapure water, filtered, lyophilized, and used for subsequent bioactivity testing.

[0131] Synthesis of β-peptide polymers with terminal thiol groups

[0132] Example 2

[0133] (2,5-dioxapyrrolidone-1-yl)2-triphenylmethylthioacetate and hexamethyldisilamide lithium salt co-initiated the preparation of random β-amino acid copolymers (NM and CH) using β-lactam monomers NM and CH. x :CH y )

[0134]

[0135] Taking a monomer ratio of x:y = 4:6 as an example, in a nitrogen-protected glove box, 1.2 mL (0.2 M) of β-lactam monomer NM solution and 1.8 mL (0.2 M) of β-lactam monomer CH solution were added to a 10 mL reaction flask. The reaction solvent was anhydrous DMAc, and the mixture was stirred. 0.3 mL (0.05 M) of (2,5-dioxapyrrolidone-1-yl)2-triphenylmethylthioacetic acid ester solution was added to the reaction flask, followed by 0.3 mL (0.15 M) of LiHMDS solution. The reaction was maintained with stirring for 4 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with 10 μL of methanol. The reaction solution was transferred to a 50 mL centrifuge tube, and an equal volume of THF solvent was added, followed by pre-cooled petroleum ether for precipitation. The THF dissolution-PE precipitation step was repeated three times to obtain a polymer with protected amino groups on the side chains. The polymer was then deprotected by adding 2 mL of trifluoroacetic acid and 5% triethylsilane overnight to remove Boc protection from the side-chain amino groups and Trt protection from the terminal groups. After overnight incubation, most of the deprotected solution was blown away with a purge pump, dissolved in a small amount of methanol, and precipitated with 45 mL of methacrylate-methyl tert-butyl ether. The dissolution-precipitation process was repeated three times, and the remaining solvent was removed by pumping out the remaining solvent. The product was then redissolved in ultrapure water and freeze-dried to obtain the deprotected random β-amino acid copolymer (yield 80%). In this example, the polymer showed a NM monomer ratio of 40% and a CH monomer ratio of 60%. Characterization by 1H NMR showed that the polymer had a DP of 40. This polymer was named 40:60NM:CH, and other polymers in this series with similar ratios were named accordingly.

[0136] Example 3

[0137] Random β-amino acid copolymers (NM) were prepared from β-lactam monomers NM and β-lactam monomers CP. x :CP y )

[0138]

[0139] The implementation method is the same as in Example 2, taking a monomer ratio of x:y = 4:6 as an example. The difference is that 1.2 mL NM (0.2 M) and 0.8 mL CH (0.2 M) are replaced with 1.2 mL NM (0.2 M) and 0.8 mL CP (0.2 M). The yield of the deprotected random β-amino acid copolymer is 82%. 1 ¹H NMR characterization showed that DP = 42.

[0140] Example 4

[0141] Random β-amino acid copolymers (NM) were prepared from β-lactam monomer NM and β-lactam monomer CO. x :CO y)

[0142]

[0143] The implementation method is the same as in Example 2, taking a monomer ratio of x:y = 4:6 as an example. The difference is that 1.2 mL NM (0.2 M) and 0.8 mL CH (0.2 M) are replaced with 1.2 mL NM (0.2 M) and 0.8 mL CO (0.2 M). The yield of the deprotected random β-amino acid copolymer is 79%. 1 ¹H NMR characterization showed that DP = 38.

[0144] Preparation of α / β-amino acid polymers

[0145] Example 5

[0146] Random α / β-amino acid copolymers were prepared from N-ε-tert-butoxycarbonyl-L-lysine-N-carboxyl intracyclic anhydride and DL-β-glycine-N-carboxylthiocarbonyl intracyclic anhydride (β3-HGNTA).

[0147]

[0148] In a nitrogen-protected glove box, taking a monomer ratio of x:y = 6:4 as an example, weigh N-ε-tert-butoxycarbonyl-L-lysine-N-carboxylic acid anhydride and DL-β-glycine-N-carboxythiocarbonyl acid anhydride. Using dry N,N-dimethylformamide as a solvent, mix 1.2 mL of N-ε-tert-butoxycarbonyl-L-lysine-N-carboxylic acid anhydride (0.2 M) and 0.8 mL of DL-β-glycine-N-carboxythiocarbonyl acid anhydride (0.2 M) in a reaction flask and stir with a magnetic stirrer. Weigh out the initiator p-tert-butylbenzylamine and prepare a 0.2M solution. Add 100 μL of this solution rapidly to the reaction flask. Stir the reaction mixture at room temperature in a glove box for 3 days. Remove the reaction solution from the glove box and add 45 mL of cold petroleum ether. After a white flocculent precipitate forms, collect the precipitate by centrifugation, dissolve it in tetrahydrofuran (1 mL), and precipitate again with cold petroleum ether. Repeat this process three times to obtain a polymer with a protecting amino group on the side chain. Gel permeation chromatography (GPC) identified the molecular weight (Mn) of the polymer with the protecting group as 3300 and the molecular weight distribution (PDI) as 1.14. Trifluoroacetic acid (2 mL) was then added to the polymer, and the mixture was shaken for 2 hours to remove the protecting groups. After most of the trifluoroacetic acid was blown away, 50 mL of methyl tert-butyl ether was added to precipitate a white precipitate, which was collected by centrifugation. The precipitate was then dissolved in methanol (1 mL) and precipitated again with 50 mL of ethyl ether. This process was repeated three times. The remaining solvent was then removed by pumping out the solvent. The samples were then dissolved in ultrapure water (5 mL) and finally lyophilized to obtain the deprotected random α / β-amino acid copolymer (yield 72.5%, DP = 20 as determined by 1H NMR).

[0149] Example 6

[0150] Random α / β-amino acid copolymers were prepared from N-ε-tert-butoxycarbonyl-L-lysine-N-carboxyl intracyclic anhydride and β-2,3-cyclohexyl-N-carboxylthiocarbonyl intracyclic anhydride (β2,3-CHNTA).

[0151]

[0152] The experimental method was the same as in Example 5, using a monomer ratio of x:y = 4:6 as an example. The difference was that 1 mL of N-ε-tert-butoxycarbonyl-L-lysine-N-carboxyl cyclic anhydride (0.2 M) and 1 mL of DL-β-glycine N-carboxylthiocarbonyl cyclic anhydride (0.2 M) were replaced with 0.8 mL of N-ε-tert-butoxycarbonyl-L-lysine-N-carboxyl cyclic anhydride (0.2 M) and 1.2 mL of β2,3-cyclohexyl N-carboxylthiocarbonyl cyclic anhydride (0.2 M), and dry tetrahydrofuran was used as the solvent. The reaction ended after 4 days. Gel permeation chromatography (GPC) identified the molecular weight (Mn) of the polymer with protecting side chains as 3300, and the molecular weight distribution (PDI) as 1.14. After deprotection, a random α / β-amino acid copolymer was obtained (yield 75%, DP = 20 as determined by 1H NMR).

[0153] Preparation of oxazoline polymers

[0154] Example 7

[0155] Preparation of oxazoline copolymers from N-ε-tert-butoxycarbonyl-2-(aminopropyl)oxazoline and 2-(cyclohexyl)oxazoline (GABA) x CH y )

[0156]

[0157] In a nitrogen-protected glove box, using a monomer ratio of x:y = 5:5 as an example, dry N,N-dimethylacetamide was used as the solvent. 1.0 mL of N-ε-tert-butoxycarbonyl-2-(aminopropyl)oxazoline (1.0 M) and 1.0 mL of 2-(cyclohexyl)oxazoline (1.0 M) were mixed and stirred, followed by the addition of 3-triphenylmethylpropane bromide as an initiator. The reaction was carried out at 140 °C for 18 hours. After cooling to room temperature, 45 mL of cold petroleum ether was added. After a white flocculent precipitate formed, the mixture was centrifuged, and the supernatant was discarded. The dissolution-precipitation-centrifugation process was repeated three times to obtain the amino-protected polypeptide copolymer. Gel permeation chromatography (GPC) identified the polymer with protecting side chains as having a molecular weight (Mn) of 4822 and a molecular weight distribution (PDI) of 1.14. After adding 2 mL of trifluoroacetic acid to the polymer and shaking for 2 hours to remove the protecting group, most of the trifluoroacetic acid was blown away. Then, 50 mL of methacin was added to precipitate a white precipitate, which was collected by centrifugation. The precipitate was then dissolved in 1 mL of methanol and precipitated again with 50 mL of methacin. This process was repeated three times. The remaining solvent was then removed by pumping out the solvent. The samples were then dissolved in 5 mL of ultrapure water and finally lyophilized to obtain the deprotected oxazoline copolymer with a yield of 83%. Characterization by 1H NMR showed a DP of 30.

[0158] Example 8

[0159] Preparation of oxazoline polymers from N-ε-tert-butoxycarbonyl-2-(aminopropyl)oxazoline and 2-(isobutyl)oxazoline (GABA) x iBu y )

[0160]

[0161] The experimental method was the same as in Example 7, using a monomer ratio of x:y = 5:5 as an example. The difference was that 1.0 mL of N-ε-tert-butoxycarbonyl-2-(aminopropyl)oxazoline (1.0 M) and 1.0 mL of 2-(cyclohexyl)oxazoline (1.0 M) were replaced with 1.0 mL of N-ε-tert-butoxycarbonyl-2-(aminopropyl)oxazoline (1.0 M) and 1.0 mL of 2-(isobutyl)oxazoline (1.0 M). The molecular weight (Mn) of the polymer with protecting groups on the side chains was identified as 4800 by gel permeation chromatography (GPC), and the molecular weight distribution (PDI) was 1.15.

[0162] Example 9

[0163] Glass surface modification polymer

[0164] Using glass as a substrate, the surface was irradiated with ultraviolet ozone for 25 minutes to introduce active oxygen groups. Then, it was immersed overnight in a 2% toluene solution of 3-aminopropyltriethoxysilane for surface amination. The amination-treated surface was vacuum annealed at 100°C for 2 hours to firmly bond the surface groups to the glass surface. Next, a polymer containing thiol groups was grafted onto the glass surface using maleic anhydride-polyethylene glycol-N-hydroxysuccinimide (with a PEG8 superhydrophilic layer as a substrate to prevent non-specific protein adhesion). Figure 1 As shown, the successful modification of the glass surface by the polymer was demonstrated by using X-ray photoelectron spectroscopy and contact angle detection.

[0165] Example 10

[0166] Screening of human umbilical vein endothelial cell adhesion and proliferation on polymer-modified surfaces

[0167] Step (1), Cell Culture. Human umbilical vein endothelial cells were cultured in culture dishes using ECM medium containing 5% fetal bovine serum (FBS), 1% penicillin / streptomycin, and 1% endothelial growth factor supplement. The medium was changed every two days until 80-90% confluence was achieved. Cells from passages 3-10 were used for in vitro experiments.

[0168] Step (2), cell adhesion screening. Cells were digested with 800 μL of 0.25% trypsin, supplemented with 2 mL of serum-containing culture medium, and centrifuged at 1400 rpm and 4°C for 4 min. The obtained cells were rehydrated with culture medium, and 10 μL was transferred to a cell counting chamber for cell counting. The cell seeding density on the polymer-modified surface was 11 × 10⁻⁶ cells / mL. 4 Cells / mL were cultured for 1 day and 3 days, and then microscopic images were taken to detect cell adhesion morphology.

[0169] Screening experiment results as follows Figures 2-1 to 2-3 As shown, endothelial cells adhered to all polymer-modified surfaces to a certain extent, and the spreading area increased with the increase of the proportion of hydrophobic monomers, indicating improved adhesion. For the three groups of polymers with different hydrophobic monomers, the adhesion of endothelial cells on the NMCH series modified surfaces was better than that on the NMCP and NMCO series modified surfaces. The β polymer NM:CH 40:60 was selected for further research.

[0170] Step (3), cell proliferation detection. Using 10% μL of MTT cell viability assay solution (5 mg / ml), incubate at 37°C in the dark for 4 hours, and detect the absorbance at 570 nm wavelength using an ELISA reader to detect cell proliferation after 1 day and 3 days.

[0171] HUVEC cell proliferation MTT such as Figures 4-1 to 4-3As shown, it is evident that the polymer NM:CH40:60 of the present invention selectively promotes the proliferation of HUVECs.

[0172] Example 11

[0173] Screening of human umbilical artery smooth muscle cell adhesion and proliferation on polymer-modified surfaces

[0174] The experimental method was the same as in Example 13, except that the human umbilical vein endothelial cells were replaced with human umbilical artery smooth muscle cells. The culture medium was replaced with SMCM medium containing 2% fetal bovine serum (FBS), 1% penicillin / streptomycin, and 1% smooth muscle cell growth factor supplement.

[0175] The adhesion screening results of HUASMC are as follows: Figures 3-1 to 3-3 As shown, using RGD (arginine-glycine-aspartic acid tripeptide) peptide as a positive control, it can be seen that HUASMC cells on the α polymer-modified surface ( Figure 3-1 Although the spreading area of ​​smooth muscle cells increases with the increase of the proportion of hydrophobic groups, Lys:Nle = 60:40 and Lys:Nle = 50:50 still exhibit excellent selectivity in promoting endothelial cell adhesion and inhibiting smooth muscle cell adhesion. On β-polymer modified surfaces ( Figure 3-2 Smooth muscle cells, compared to those on RGD-modified surfaces, exhibited spherical aggregation, poor spreading, and a tendency to detach. On oxazoline polymer-modified surfaces (… Figure 3-3 Smooth muscle cells showed a contractile trend on surfaces modified with GABA:CH = 80:20, 70:30, and 30:70, indicating a certain selective inhibitory effect. The MTT results are as follows: Figures 5-1 to 5-3 As shown, the surface modified by polymer NM:CH 40:60 significantly inhibited the proliferation of HUASMC.

[0176] Example 12

[0177] Polymer-modified surface co-culture of human umbilical vein endothelial cells and human umbilical artery smooth muscle cells

[0178] Human umbilical vein endothelial cells and human umbilical artery smooth muscle cells were incubated with Cell Tracker Green CMFDA (5 mM) and Orange CMTMMR (5 mM), respectively (37°C, 20 min). After washing with PBS, the cells were incubated for 30 min with dye-free medium (ECM:SMCM 1:1 blend). Both cell types were then digested with 0.25% trypsin, centrifuged at 4°C for 4 min, resuspended in dye-free medium, and the cell density was adjusted to 10 × 10⁶ cells / mL. 4The two cell types were mixed at a 1:1 planting density and seeded onto the polymer-modified surface of this invention. The mixture was cultured at 37°C with 5% CO2. Morphological and numerical differences between the two cell types were observed using a fluorescence microscope at 6 h and 12 h. The experimental results are as follows: Figure 6 As shown, red represents HUASMC and green represents HUVEC. RGD (arginine-glycine-aspartic acid tripeptide) and REDV (arginine-glutamic acid-aspartic acid-valine tetrapeptide) were used as positive controls. It can be seen that, compared with HUASMC, the polymer-modified surface of the present invention has a clear selective function of promoting HUVEC adhesion.

[0179] Example 13

[0180] Metal surface modified polymer

[0181] Hexamethylenediamine (2.44 mg / ml) and dopamine (1 mg / ml) were blended, and tris(hydroxymethyl)aminomethane hydrochloride (pH = 8.5) was used as the solvent. A 316 low-carbon stainless steel (316LSS) substrate was immersed in the above solution overnight for surface amination. The amination surface was then subjected to the same experimental method as in Example 12.

[0182] Example 14

[0183] Cell migration on polymer-modified surfaces

[0184] A 316 low-carbon stainless steel substrate was bent at 90°, with one side modified with the polymer of the present invention and the other side being the bare stainless steel surface. A 10×10 slurry was then seeded onto the bare stainless steel surface. 4 Human umbilical vein endothelial cells or human umbilical artery smooth muscle cells were cultured at a density of 200 μL / mL to form a dense monolayer. The sample was then inverted 90° so that the polymer-modified surface was parallel to the well plate. Fresh culture medium was added to induce cell migration from the bare metal to the polymer-modified surface. After 24 hours of culture, migration was observed using a fluorescence microscope, and the migration distance was calculated using ImageJ. The experimental results are as follows: Figure 7 As shown, compared with the non-selective 316LSS stainless steel surface and the RGD-modified surface, the polymer-modified surface exhibits superior selective migration capabilities and has a greater promoting effect on HUVEC migration than the REDV-modified surface.

[0185] Example 15

[0186] Study on implantation of polymer-modified vascular stent mimics into rats

[0187] The degree of endothelialization and proliferation on the modified surface was evaluated using a rat abdominal aorta implantation model with polymer-modified steel wire. The implanted samples were: group a, bare steel wire (316LSS); and group b, polymer-modified steel wire (n=6).

[0188] The steel wire and all reagents were sterile, and the preparation process was also carried out under sterile conditions. The specific preparation process is as follows:

[0189] (1) Preparation of polymer-modified steel wire: The modification method is the same as in Example 16;

[0190] (2) Storage: Store in a refrigerator at 4℃.

[0191] Twelve male SD rats (weighing more than 250g) were purchased from Shanghai Silex Company and implanted into two pre-prepared samples, with six parallel samples in each sample group.

[0192] Animal experiments were conducted strictly in accordance with laboratory animal care and usage guidelines. The surgical procedure was as follows:

[0193] (1) Animal anesthesia: Each rat was anesthetized by injecting one milliliter of sodium pentobarbital (200g / ml) into the peritoneum;

[0194] (2) Surgical preparation: After waiting for a period of time, the rat was completely anesthetized. The hair on the abdomen was shaved off with a shaver and the surgical site was disinfected with iodine.

[0195] (3) Sample implantation: Using surgical scissors, dissect the abdomen along the midline. Push the abdominal organs to the periphery with cotton balls soaked in saline solution, cover with gauze soaked in saline solution to prevent the body fluid from drying out, and fix the wound with clamps. Carefully dissect with cotton swabs to expose the arteries and veins, then carefully separate the abdominal aorta and veins, and inject heparin to cleanse the arteries. The vascular stent is inserted into the aorta with a heparin-cleaned needle, inserted from the lower end and exited from the upper end, leaving the steel wire sample inside the blood vessel. The steel wire is sutured at three ends with 9-0 vascular sutures to fix the steel wire to the inner wall of the blood vessel.

[0196] (4) Surgical suturing: The abdominal wound was sutured, the wound was disinfected with iodine, a dressing was applied, and the animal was placed on an electric blanket. When it was about to wake up, it was transferred to a cage for observation for 24 hours.

[0197] (5) Feeding: To prevent infection and inflammation of the mouse wounds, administer antibiotics after the mouse recovers and continue feeding.

[0198] One month later, the wire and surrounding blood vessels were removed, fixed with 4% paraformaldehyde, and embedded in paraffin for tissue section observation and analysis. The tissue section results are as follows: Figure 8 As shown, the implantation of polymer-modified vascular stent mimics significantly reduced tissue proliferation, with a marked decrease in the proliferative area.

[0199] In summary, the polymer of the present invention selectively promotes endothelial cell adhesion and growth, making it highly suitable for modification (coating, adsorption, chemical grafting, etc.) onto the surface of interventional / implantable materials that require endothelialization, thereby enabling them to function more effectively and to remain in the patient's body more safely and for longer.

[0200] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. Use of a polymer, or a pharmaceutically acceptable salt thereof, in the manufacture of a cardiovascular implant material for promoting in situ endothelialization, wherein, The polymer comprises a segment selected from Formula I or Formula II: I、 II wherein in Formula I, n is a positive integer from 1 to 100; r1 and r2 are each independently 1 or 2; one of R1is -L1R a wherein L1is substituted or unsubstituted C1-C8alkylene; R a is selected from the group consisting of -NR b R b , -N(R b )3 + ; and each R b is independently selected from the group consisting of hydrogen, substituted or unsubstituted C1-C4alkyl; the remainder of the groups are H; and a the remainder of the groups are H; and the remainder of the groups are H; and the remainder of the groups are H; and the remainder of each R2 is independently selected from the group consisting of H, C1-C8 alkyl, or C1-C6 alkyl-Rc, wherein Rcis phenyl; or two R2 together with the carbon atom to which they are attached form a substituted or unsubstituted C3-C8 cycloalkyl; in Formula II, n is a positive integer from 1 to 100; R3is -L1R a wherein L1is selected from the group consisting of a bond, C1-C8alkylene; R a is selected from the group consisting of -NR b R b , -N(R b )2 + ; and each R b is independently selected from the group consisting of hydrogen, C1-C4alkyl; R4 is selected from the group consisting of H, C1-C8 alkyl, or C3-C8 cycloalkyl; in each formula, x is independently from 10 to 90, y is independently from 10 to 90; and x+y=100.

2. Use according to claim 1, characterized in that, , -L1R a independently -C2-C6alkyl-R a .

3. Use according to claim 1, characterized in that, each R is independently selected from the group consisting of -NH2, -NH3 a , -N(CH3)3 + , -N(C2H5)3 + , and -N(C3H7)3 + .

4. The use according to claim 1, characterized in that, r1 and r2 are 1, or r1 and r2 are 2, or r1 is 1 and r2 is 2, or r1 is 2 and r2 is 1.

5. The use according to claim 1, characterized in that, at least one R2 group is selected from the group consisting of C1-C6 alkyl, or two R2 together with the carbon atom to which they are attached form a substituted or unsubstituted C3-C6 cycloalkyl.

6. The use according to claim 1, characterized in that, The polymer is selected from the group consisting of: in each formula, x is independently from 10 to 90, y is independently from 10 to 90; and x+y=100; n is a positive integer from 1 to 100.

7. The use according to claim 1, characterized in that, The polymer is coated or covalently bonded to at least a portion of the surface of the substrate.

8. Use according to claim 7, characterized in that, The substrate is selected from the group consisting of inorganic non-metallic biomaterials, metallic biomaterials, natural polymeric materials, synthetic polymeric materials, or combinations thereof.

9. The use according to claim 7, characterized in that, The substrate is selected from the group consisting of bioceramics, bioglasses, graphene, bone cement, medical carbon materials, stainless steel, cobalt-based and titanium-based alloys, shape memory alloys, silver, platinum, tantalum, niobium, zirconium, palladium, hyaluronic acid, chitosan, alginic acid, cellulose, collagen, gelatin, polyether ether ketone, polycaprolactone, polylactic acid, polycarbonate, polyurethane, poly anhydride, polydimethylsiloxane, polymethyl methacrylate, polyphosphazene, polyamide, polyethylene, polypropylene, polytetrafluoroethylene, polyethylene terephthalate, or composites or assembled materials thereof.

Images