CN120271814A - Protein-like particles and preparation method thereof - Google Patents
Protein-like particles and preparation method thereof Download PDFInfo
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- CN120271814A CN120271814A CN202510453306.0A CN202510453306A CN120271814A CN 120271814 A CN120271814 A CN 120271814A CN 202510453306 A CN202510453306 A CN 202510453306A CN 120271814 A CN120271814 A CN 120271814A
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
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- C08J2377/00—Characterised by the use of polyamides obtained by reactions forming a carboxylic amide link in the main chain; Derivatives of such polymers
- C08J2377/04—Polyamides derived from alpha-amino carboxylic acids
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Abstract
本发明提供了一种仿蛋白颗粒及其制备方法，属于纳米材料技术领域。本发明提供的仿蛋白颗粒中C链段上的双键可通过点击化学接阳性药或荧光基团，羧基可以通过DCC偶联连接靶向肽、含羟基或者氨基的阳性药等，从而得到具有磁性的携带药物分子、荧光基团、靶向肽的仿蛋白颗粒，其中，所述具有亲水性的A链段使其在生物体内递送，B具有磁性，可穿过一些具有阻力的组织液片段，从而进入到常规药物无法进入的部位进行治疗，同时靶向肽可以使其与体内蛋白结合，使该仿蛋白颗粒更易进入生物体内，有效提高了药物在病变部位的蓄积浓度，提高疗效；所述仿蛋白颗粒尺寸小，对人体安全性高，毒副作用小。
The present invention provides a protein-mimicking particle and a preparation method thereof, belonging to the technical field of nanomaterials. The double bonds on the C segment in the protein-mimicking particle provided by the present invention can be connected to a positive drug or a fluorescent group through click chemistry, and the carboxyl group can be connected to a targeting peptide, a positive drug containing a hydroxyl group or an amino group through DCC coupling, so as to obtain a magnetic protein-mimicking particle carrying a drug molecule, a fluorescent group, and a targeting peptide, wherein the hydrophilic A segment enables it to be delivered in vivo, and B has magnetism and can pass through some tissue fluid fragments with resistance, so as to enter a part that conventional drugs cannot enter for treatment, and at the same time, the targeting peptide can make it bind to the protein in the body, so that the protein-mimicking particle is easier to enter the body, effectively increasing the accumulation concentration of the drug in the lesion site and improving the efficacy; the protein-mimicking particle is small in size, highly safe for the human body, and has small toxic and side effects.
Description
Technical Field
The invention relates to the technical field of nano materials, in particular to a protein-like particle and a preparation method thereof.
Background
Proteins are important chemical substances in natural organisms as scaffolds and main substances constituting human tissue organs. Proteins in organisms are assembled from folds of polypeptide chains, which, due to their complex sequence and diversity of chemical functions, can produce a structurally defined folded chain with high specificity, whereas polypeptides have the same backbone repeat sequence as proteins. In the 40 s of the 20 th century, chemists and biochemists from around the world began research projects in the emerging field of synthetic polypeptides. Because of the limited variety of side chain functional groups and the limited variety of natural amino acids, researchers prepare artificially synthesized polyamino acids by a chemical method of ring-opening polymerization of alpha-amino acid-N-carboxylic acid hydride (NCA) monomers, can contain various functional groups, simultaneously rapidly prepare high molecular chain polypeptides in high yield and large scale, simulate the folding of proteins in human bodies, and have the characteristics of good biocompatibility, good biological metabolism capability, multifunction, low toxicity and the like. Thus, polyamino acids may be considered as ideal choices for drug delivery.
In addition, the magnetic nano particles can be controlled by utilizing a magnetic field, have interesting characteristics of uniform size, high surface area, biocompatibility, superparamagnetism, adsorption kinetics, magnetic moment and the like, are widely applied to the fields of thermotherapy, targeted drug delivery systems, imaging, biomolecule extraction and the like, and become an important tool for tumor treatment. The synthesis method of magnetic nanoparticles such as iron oxide nanoparticles includes wet chemical or "bottom-up" approaches such as hydrothermal, solvothermal, sol-gel, coprecipitation, flow injection synthesis, electrochemical, laser pyrolysis techniques, and the like. The magnetic nano particles are coated with biological functional molecules such as antibodies, ligands or receptors to enable the magnetic nano particles to have high-affinity interaction with biological entities by modifying the surfaces of the magnetic nano particles, so that the magnetic nano particles are improved in biocompatibility, poor biodegradability, chemical instability in physiological environments and the like, and a controllable marking means is provided, and high selectivity and high sensitivity are provided for a plurality of biological applications. The existing magnetic nano particles and similar protein-like particle metals have the defects of excessively high net content and lack of degradable organic frameworks, complex synthesis steps, strict control of reaction conditions and high production cost, so that the difficulty of mass production is high, and the feasibility of mass production is limited.
Disclosure of Invention
The invention aims to provide a protein-like particle and a preparation method thereof, wherein the protein-like particle has an asymmetric structure and excellent motility, is provided with multiple double bonds, can be used as a nano carrier and connected with a positive drug through click chemistry, has good biocompatibility due to a multiple carboxyl structure, can be connected with a targeting peptide through DCC coupling, has targeting property, has small size, can be discharged out of the body through kidneys in vivo, and has high safety to human bodies and small toxic and side effects.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a protein-like particle, which has the structure of A-B-C, wherein the A chain segment is a hydrophilic linear polymer chain, B is a spherical or spheroidic nanoparticle, and the C chain segment is a linear polymer chain with double bonds and carboxyl on the side chain;
The number average molecular weight of the A chain segment and the C chain segment is 50-5000000 independently.
Preferably, the hydrodynamic diameter of the spherical or spheroid nanoparticle B is 1-1000 nm, and the C chain segment is a polyamino acid chain segment with double bonds and carboxyl groups on the side chains.
The invention also provides a preparation method of the protein-like particles, which comprises the following steps:
Mixing a linear high molecular compound with hydrophilicity, a first NCA monomer and a first solvent, and carrying out ring-opening polymerization reaction to obtain a first block polymer;
Mixing the first block polymer, a second NCA monomer and a second solvent, and carrying out polymerization reaction to obtain a second block polymer;
Mixing the second block polymer, the metal carbonyl compound and the third solvent, and performing complex crosslinking reaction to obtain single-chain nano particles;
And sequentially carrying out post-treatment and deprotection reaction on the single-chain nano particles to obtain the protein-like particles.
Preferably, the linear polymer compound with hydrophilicity is at least one of polyethylene glycol, polyacrylic acid, polymethacrylic acid, polyvinyl alcohol, polyethyleneimine, sodium polyacrylate, polyamino acid and polyacrylamide.
Preferably, the first NCA monomer is but-3-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, prop-2-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 4- (prop-2-yn-1-oxy) -benzyl-L-glutamate-N-carboxy-inner anhydride, 2-methyl-but-3-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 2-dimethyl-but-3-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, pent-4-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 1-methyl-but-3-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 2-ethynyl-but-3-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 3-methyl-pent-4-yn-1-yl-L-glutamate-N-carboxy-inner anhydride, 2-methyl-pent-4-yl-L-carboxy-glutamate-N-inner anhydride, at least one of 1-methyl-pent-4-yn-1-yl-L-glutamic acid-N-carboxyinner anhydride.
Preferably, the second NCA monomer is at least one of a double bond-containing NCA monomer, a tert-butyl group-containing NCA monomer, a halogen-containing functional group-containing NCA monomer;
the NCA monomer containing double bonds is at least one of butyl-3-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride, propyl-2-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride, 4-ethyleneoxy benzyl-L-glutamic acid-N-carboxyl internal anhydride and 2-methyl-butyl-3-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride;
the NCA monomer containing tert-butyl ester group is at least one of tert-butyl ester-L-glutamic acid-N-carboxyl inner anhydride, methyl ester-L-glutamic acid-N-carboxyl inner anhydride, benzyl ester-L-glutamic acid-N-carboxyl inner anhydride and isobutyl ester-L-glutamic acid-N-carboxyl inner anhydride;
The NCA monomer containing the halogen functional group is at least one of 3-chloro-prop-1-yl-L-glutamic acid-N-carboxyl inner anhydride, 4- (chloromethyl) -1-benzyl-L-glutamic acid-N-carboxyl inner anhydride, 2-trichloro-acetic-1-yl-L-glutamic acid-N-carboxyl inner anhydride and 4- (bromomethyl) -1-benzyl-L-glutamic acid-N-carboxyl inner anhydride.
Preferably, the metal carbonyl compound is at least one of Co2(CO)8、Fe2(CO)9、Fe(CO)5、Ni(CO)4、Mn2(CO)10、Re2(CO)10、V(CO)6、Co4(CO)12、Pt(CO)4、Pt(CO)3、Pt(CO)2、Pd(CO)4、Mo(CO)6、W(CO)6、Cr(CO)6、Ru3(CO)12、 cisplatin dichloride dicarbonyl.
Preferably, the solvent for the complex crosslinking reaction is at least one of o-dichlorobenzene, chlorobenzene, N, N-dimethylformamide, N, N-dimethylacetamide, tetrahydrofuran, dimethyl phthalate, dichloromethane, chloroform, 1, 2-dichloroethane, 1, 2-tetrachloroethane and toluene.
Preferably, the post-treatment is at least one of heating, catalytic hydrogenation, microwave treatment, high-energy electron beam treatment, gamma ray irradiation treatment, nuclear radiation treatment.
Preferably, when the post-treatment is heating, the heating temperature is 80-300 ℃, and the solvent used for heating is at least one of o-dichlorobenzene, N, N-dimethylformamide, N, N-dimethylacetamide, dimethyl phthalate and acetophenone.
The invention provides a protein-like particle, which has the structure of A-B-C, wherein an A chain segment is a linear polymer chain with hydrophilicity, B is spherical or spheroidic nano particles, a C chain segment is a linear polymer chain with a side chain containing double bonds and carboxyl, and the number average molecular weight of the A chain segment and the number average molecular weight of the C chain segment are respectively 50-5000000. The C=C on the tail end C chain segment of the protein-like particle provided by the invention can be connected with a positive drug or a fluorescent group through click chemistry, and a carboxyl can be connected with a targeting peptide, a positive drug containing hydroxyl or amino and the like through DCC coupling, so that the protein-like particle which has the size of 5-25 nm and magnetic property and carries drug molecules, fluorescent groups and targeting peptides is obtained. The invention has the innovation point that the synthesized imitated protein particle size is smaller than that of a common nano medicine-carrying material, the linear polymer chain A with hydrophilicity can enable the imitated protein particle to be delivered in a living body, the B has magnetism and can pass through a plurality of tissue fluid fragments with resistance so as to enter into a part which cannot be entered by a conventional medicine for treatment, meanwhile, the targeting peptide can enable the imitated protein particle to be combined with internal proteins, so that the imitated protein particle can enter into the living body more easily, the accumulation concentration of the medicine in a lesion part is effectively improved, the C chain segment has a multi-carboxyl structure, the C chain segment has good biocompatibility, electrostatic adsorption between COO-and the protein is easily shielded by the ion effect in a solution, the surface adsorption quantity is reduced, the characteristics of good biodegradability and low toxicity are achieved, meanwhile, the imitated protein particle has an asymmetric structure, the A chain segment is taken as a molecular tail, the magnetic nano particle B is taken as a head part, the like the tissue fluid, the imitated protein particle can enter into the part through peristalsis of the self so that the medicine can enter into the lesion part more easily, the tumor part easily, the accumulation is improved, in addition, the imitated protein particle is easy to be concentrated, the dimension is easy, the C chain segment and the small amino acid can be easily discharged from the human body in the aspect of high in the human body, and has a small-scale and has a good toxicity and a good toxic effect on the aspect when applied to the human body, and has a small-scale and a small toxicity on the human body.
The invention selects polyamino acid as a framework, has good biocompatibility, can be compatible with tissues and cells in organisms, reduces immune reaction and toxicity, has good biodegradability, can be decomposed into harmless micromolecules through enzyme degradation or hydrolysis, avoids long-term accumulation in the bodies and reduces the toxicity problem in long-term use, has the advantages of high efficiency, controllability, flexibility and the like through the open-loop polymerization of alpha-amino acid-N-carboxylic anhydride (alpha-AminoAcid-N-Carboxyanhydrides, NCA) monomer, can synthesize various polyamino acid materials with adjustable structures and various functions, can accurately regulate and control the polymerization conditions through accurately controlling the polymerization reaction condition, can still obtain polymers with high molecular weight under low polymerization degree, has high synthesis efficiency, can obtain polymers with high yield in shorter time, and is especially suitable for large-scale production.
Drawings
FIG. 1 is a schematic representation of the structures of a first NCA monomer and a second NCA monomer according to the present invention, wherein FIG. 1 a-1 is but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-2 is prop-2-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-3 is 4- (prop-2-yn-1-oxy) -benzyl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-4 is 2-methyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-5 is 2, 2-dimethyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-6 is pent-4-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, FIG. 1 a-7 is 1-methyl-but-3-yn-1-yl-L-carboxyinternal anhydride, FIG. 1-N-ethynyl-2-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, a-9 in FIG. 1 is 3-methyl-pent-4-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, a-10 in FIG. 1 is 2-methyl-pent-4-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, a-11 in FIG. 1 is 1-methyl-pent-4-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, b-1 in FIG. 1 is but-3-en-1-yl-L-glutamic acid-N-carboxyinternal anhydride, b-2 in FIG. 1 is prop-2-en-1-yl-L-glutamic acid-N-carboxyinternal anhydride, b-3 in FIG. 1 is 4-ethyleneoxybenzoyl-L-glutamic acid-N-carboxyinternal anhydride, b-4 in FIG. 1 is 2-methyl-but-3-en-1-yl-L-glutamic acid-N-carboxyinternal anhydride, c-1 in FIG. 1 is tert-butyl-L-glutamic acid-N-carboxyinternal anhydride, c-2 in FIG. 1 is methyl-2-en-1-yl-L-glutamic acid-N-c-carboxyinternal anhydride, c-N-carboxyinternal anhydride in FIG. 1-c-4-carboxyi-glutamic acid-N-carboxyinternal anhydride, d-1 in FIG. 1 is 3-chloro-prop-1-yl-L-glutamic acid-N-carboxyinner anhydride, d-2 in FIG. 1 is 4- (chloromethyl) -1-benzyl-L-glutamic acid-N-carboxyinner anhydride, d-3 in FIG. 1 is 2, 2-trichloro-prop-1-yl-L-glutamic acid-N-carboxyinner anhydride, d-4 in FIG. 1 is 4- (bromomethyl) -1-benzyl-L-glutamic acid-N-carboxyinner anhydride;
FIG. 2 is a TEM image of single-stranded nanoparticles and protein-like particles prepared in example 1 of the present invention, wherein A in FIG. 2 is a single-stranded nanoparticle and B in FIG. 2 is a protein-like particle;
FIG. 3 is a block diagram of a PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer and a protein-mimetic particle prepared according to example 1 of the present invention, wherein A in FIG. 3 is a PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer and B in FIG. 3 is a protein-mimetic particle;
FIG. 4 is a synthetic route of the protein-like particles prepared in example 1 of the present invention;
FIG. 5 is a hydrogen spectrum of a first block polymer PEO 5k-Alk30 prepared in example 1 of the present invention;
FIG. 6 is a hydrogen profile of PEO 5k-Alk30-Eth55-tBA52 prepared in example 1 of the present invention;
FIG. 7 is a GPC chart of the MeO-PEO 5k-NH2、PEO5k-Alk30 and PEO 5k-Alk30-Eth55-tBA52 of the present invention.
Detailed Description
The invention provides a protein-like particle, which has the structure of A-B-C, wherein the A chain segment is a hydrophilic linear polymer chain, B is a spherical or spheroidic nanoparticle, and the C chain segment is a linear polymer chain with double bonds and carboxyl on the side chain;
The number average molecular weight of the A chain segment and the C chain segment is 50-5000000 independently.
In the present invention, the raw materials used are all conventional commercial products in the art unless otherwise specified.
In the present invention, the spherical or spheroid nanoparticle B is preferably a metal nanoparticle, and more preferably consists of at least one of Co, fe, ni, mn, re, V, co, pt, pd, mo, W, cr, ru and platinum. In the invention, after the spherical or spheroid nano-particles B are crosslinked by alkynyl and carbonyl metal compounds, the spherical or spheroid nano-particles B are thermally decomposed after the post-treatment, CO is decomposed, and the basic metal composition is left in the polymer matrix. In the present invention, the hydrodynamic diameter of the spherical or spheroid-like nanoparticle B is preferably 1nm to 1000nm, more preferably 3 to 50nm. The invention controls the hydrodynamic diameter of B in the above range so as to more easily pass through a physiological barrier, easily enter the nucleus of a cell, go deep into a tumor and show more accumulation in tumor tissues, and the smaller the size of the nano particles is, the larger the specific surface area of the nano particles is, the surface of the nano particles is modified, thereby realizing the efficient and specific delivery of the drug, improving the bioavailability of the drug, reducing the contact of the drug with normal cells and reducing the toxic and side effects.
The invention also provides a preparation method of the protein-like particles, which comprises the following steps:
Mixing a linear high molecular compound with hydrophilicity, a first NCA monomer and a first solvent, and carrying out ring-opening polymerization reaction to obtain a first block polymer;
Mixing the first block polymer, a second NCA monomer and a second solvent, and carrying out polymerization reaction to obtain a second block polymer;
Mixing the second block polymer, the metal carbonyl compound and the third solvent, and performing complex crosslinking reaction to obtain single-chain nano particles;
And sequentially carrying out post-treatment and deprotection reaction on the single-chain nano particles to obtain the protein-like particles.
The invention mixes hydrophilic linear high molecular compound, first NCA monomer and first solvent to carry out ring-opening polymerization reaction to obtain first block polymer.
In the present invention, the linear polymer compound having hydrophilicity is preferably at least one of polyethylene glycol methyl ether amine, polyethylene glycol, polyacrylic acid, polymethacrylic acid, polyvinyl alcohol, polyethylenimine, sodium polyacrylate, polyamino acid, and polyacrylamide, and more preferably polyethylene glycol.
In the present invention, the first NCA monomer preferably contains an alkyne functional group at the gamma position of an amino acid, more preferably but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, prop-2-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, 4- (prop-2-yn-1-oxy) -benzyl-L-glutamic acid-N-carboxyinternal anhydride, 2-methyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, 2-dimethyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, pent-4-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, 1-methyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, 2-ethynyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, 3-methyl-pent-4-yl-L-yn-1-yl-N-carboxyinternal anhydride, 2-dimethyl-but-3-yn-1-yl-L-glutamic acid-N-carboxyinternal anhydride, pent-4-yl-N-carboxyinternal anhydride, 1-methyl-yn-yl-N-carboxyinternal anhydride, N-methyl-amino acid, or 2-amino acid, at least one of 1-methyl-pent-4-yn-1-yl-L-glutamic acid-N-carboxyinner anhydride.
In the present invention, the molar ratio of the hydrophilic linear polymer compound to the first NCA monomer is preferably 1 (25 to 200), more preferably 1 (30 to 150), and still more preferably 1:50. The invention controls the mole ratio of the linear high molecular compound with hydrophilicity and the first NCA monomer in the range to control the mole ratio to be 1 (25-200), can effectively control the average polymerization degree of (Glu-yne) in the polymer to be 10-100 according to the reaction ratio in the process of NCA ring-opening polymerization reaction, and avoids too short polymer chain ((Glu-yne) too few, subsequent crosslinking reaction is not easy to carry out, and finally synthesized imitated protein particles are too small) or too long (finally synthesized imitated protein particles are prevented from being too large).
In the present invention, the first block polymer is preferably a block copolymer formed by copolymerizing two or more of the first NCA monomers, or a random copolymer formed by copolymerizing three or more of the first NCA monomers, more preferably a polyethylene glycol homopolymer.
In the invention, the temperature of the ring-opening polymerization reaction is preferably room temperature, the time of the ring-opening polymerization reaction is preferably 2-4 d, and the ring-opening polymerization reaction is preferably carried out under stirring. In the present invention, the first solvent is preferably at least one of DMF, ethyl acetate, methyl acetate, isopropyl alcohol, N-butanol, N-amyl alcohol, ethylene glycol, glycerin, acetone, acetonitrile, methyl benzoate, dimethyl phthalate, methylene chloride, chloroform, 1, 2-dichloroethane, 1,2, -tetrachloroethane, nitrobenzene, methanol, ethanol, dioxane, N-Dimethylacetamide (DMA), o-dichlorobenzene, tetrahydrofuran (THF). In the invention, a polyamino acid chain segment containing a large number of alkyne functional groups grows at one end of a chain segment A (formed by a linear high molecular compound with hydrophilicity) through ring-opening polymerization reaction.
After the ring-opening polymerization reaction is completed, the present invention preferably directly feeds the ring-opening polymerization reaction product to perform the polymerization reaction.
After the first block polymer is obtained, the first block polymer, the second NCA monomer and the second solvent are mixed for polymerization reaction to obtain the second block polymer.
In the present invention, the second NCA monomer is preferably at least one of a double bond-containing NCA monomer, a tert-butyl group-containing NCA monomer, a halogen-containing functional group-containing NCA monomer;
The NCA monomer containing double bonds is preferably at least one of butyl-3-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride, propyl-2-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride, 4-ethyleneoxy benzyl-L-glutamic acid-N-carboxyl internal anhydride and 2-methyl-butyl-3-alkene-1-yl-L-glutamic acid-N-carboxyl internal anhydride;
The NCA monomer containing tert-butyl ester group is preferably at least one of tert-butyl ester-L-glutamic acid-N-carboxyl inner anhydride, methyl ester-L-glutamic acid-N-carboxyl inner anhydride, benzyl ester-L-glutamic acid-N-carboxyl inner anhydride and isobutyl ester-L-glutamic acid-N-carboxyl inner anhydride;
The NCA monomer containing halogen functional group is preferably at least one of 3-chloro-prop-1-yl-L-glutamic acid-N-carboxyl inner anhydride, 4- (chloromethyl) -1-benzyl-L-glutamic acid-N-carboxyl inner anhydride, 2-trichloro-acetic-1-yl-L-glutamic acid-N-carboxyl inner anhydride and 4- (bromomethyl) -1-benzyl-L-glutamic acid-N-carboxyl inner anhydride.
In the invention, the temperature of the polymerization reaction is preferably room temperature, the time of the polymerization reaction is preferably 1-3 d, and the polymerization reaction is preferably carried out under the condition of stirring. In the present invention, the second solvent is preferably at least one of DMF, ethyl acetate, methyl acetate, isopropyl alcohol, N-butanol, N-amyl alcohol, ethylene glycol, glycerin, acetone, acetonitrile, methyl benzoate, dimethyl phthalate, methylene chloride, chloroform, 1, 2-dichloroethane, 1,2, -tetrachloroethane, nitrobenzene, methanol, ethanol, dioxane, N-Dimethylacetamide (DMA), o-dichlorobenzene, tetrahydrofuran (THF). The invention forms a linear polymer chain C 'with double bonds and tert-butyl ester groups on one end of a polyamino acid chain segment through polymerization reaction, wherein the chain segment C' is preferably formed by ring opening homopolymerization of a second NCA monomer, random ring opening copolymerization of a plurality of second NCA monomers or sequential feeding of a plurality of first NCA monomers, and polymerization of the first NCA monomers segment by segment.
After the polymerization reaction is completed, the polymerization reaction product is preferably subjected to evaporation, redissolution, precipitation and centrifugation in sequence to obtain a second block polymer.
In the invention, the evaporation is preferably rotary evaporation, the reagent used for redissolution is preferably dichloromethane, the reagent used for precipitation is preferably diethyl ether, the centrifugation time is preferably 20-50 min, more preferably 30min, and the rotation speed of the centrifugation is preferably 4000-6000 rpm, more preferably 5000rpm.
After the second block polymer is obtained, the second block polymer, the carbonyl metal compound and the third solvent are mixed for complex crosslinking reaction to obtain the single-chain nano-particles.
In the present invention, the metal carbonyl compound is preferably at least one of Co2(CO)8、Fe2(CO)9、Fe(CO)5、Ni(CO)4、Mn2(CO)10、Re2(CO)10、V(CO)6、Co4(CO)12、Pt(CO)4、Pt(CO)3、Pt(CO)2、Pd(CO)4、Mo(CO)6、W(CO)6、Cr(CO)6、Ru3(CO)12、 cisplatin dichloride dicarbonyl.
In the present invention, the third solvent is preferably at least one of o-dichlorobenzene, chlorobenzene, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dimethyl phthalate, dichloromethane, chloroform, 1, 2-dichloroethane, 1, 2-tetrachloroethane, toluene, and more preferably dimethyl phthalate. The third solvent is used as a solvent for the complexation reaction, and is used for simultaneously dissolving the polymer and the metal carbonyl compound, and has weak coordination capacity, so that the smooth progress of the complexation crosslinking reaction is promoted.
In the invention, the temperature of the complexing crosslinking reaction is preferably room temperature, the time of the complexing crosslinking reaction is preferably 3-5 d, and the complexing crosslinking reaction is preferably carried out under the condition of stirring. The invention utilizes coordination, the polyamino acid chain segment contains a large number of alkyne functional groups to carry out complexation crosslinking reaction with the carbonyl metal compound, thereby realizing the folding of the chain segment and forming the polyamino acid chain segment with the carbonyl metal compound.
After the single-chain nano particles are obtained, the single-chain nano particles are subjected to aftertreatment and deprotection reaction in sequence to obtain the protein-like particles.
In the present invention, the post-treatment is preferably at least one of heating, catalytic hydrogenation, microwave treatment, high-energy electron beam treatment, gamma ray irradiation treatment, and nuclear radiation treatment.
In the invention, when the post-treatment is heating, the heating temperature is preferably 80-300 ℃, more preferably 100-200 ℃, and the heating time is preferably 10-16 h, more preferably 12-14 h. In the invention, the heating is preferably performed in a nitrogen gas flow, and the solvent used for the heating is preferably at least one of o-dichlorobenzene, N, N-dimethylformamide, N, N-dimethylacetamide, dimethyl phthalate and acetophenone. According to the invention, through heating, the metal carbonyl compound in the polyamino acid chain segment structure with the metal carbonyl compound is subjected to thermal decomposition reaction, and the carbon monoxide component in the metal carbonyl compound is dissociated to form metal nano particles, namely spherical or spheroidic nano particles B.
In the invention, the temperature of the deprotection reaction is preferably-50-100 ℃, more preferably 20-30 ℃, further preferably 25 ℃, the time of the deprotection reaction is preferably 12-96 h, more preferably 24-60 h, further preferably 48h, and all reagents of the deprotection reaction are preferably at least one of trifluoroacetic acid, hydrochloric acid, sulfuric acid, nitric acid, glacial acetic acid, propionic acid, perchloric acid, phosphoric acid and hydrofluoric acid. According to the invention, the tertiary butyl on the side chain of the C' chain segment structure with the side chain containing double bonds and tertiary butyl groups is removed through a deprotection reaction, so that a linear high polymer chain C with the side chain containing double bonds and carboxyl groups is formed, and finally the protein-like particle with a chain-ball-chain morphology is obtained.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The sources of some of the raw materials used in the examples of the present invention are shown in Table 1, and the sources of the instruments are shown in Table 2.
TABLE 1 partial sources of raw materials
Table 2 instrument source
According to HIGHLY EFFICIENT 'Grafting onto' aPolypeptide Backbone Using CLICK CHEMISTRY and THE SYNTHETIC tuning of clickable pH responsive cationic polypeptides and block copolypeptides which are references of Alk-NCA, reference ;Synthesis and ring-opening(Co)polymerization ofL-lysineN-carboxyanhydrides containing labile side-chain protective groups of thio-Ene Clickable Polypeptides which is Eth-NCA which is reference of tBA-NCA, three monomers of but-3-yn-1-yl-L-glutamic acid-N-carboxyanhydride (Alk-NCA), but-3-en-1-yl-L-glutamic acid-N-carboxyanhydride (Eth-NCA) and tert-butyl-L-glutamic acid-N-carboxyanhydride (tBA-NCA) which are used in the examples of the invention are sequentially synthesized, specifically as follows:
(1) The preparation method of the Alk-NCA comprises the steps of weighing 0.2g of L-glutamic acid, 3mL of 3-butyn-1-ol and 0.44mL of trimethylchlorosilane in a 25mL eggplant bottle, gradually clarifying the solution after the reaction is completed under the condition of nitrogen for 24 hours, precipitating in diethyl ether for a small amount for many times, centrifuging to obtain 0.32g of intermediate 1a as white solid, weighing 0.8g of intermediate 1a and 0.12g of triphosgene in a 25mL reaction tube, adding 11mL of ultra-dry ethyl acetate into the eggplant bottle, heating and refluxing for 5 hours under the condition of nitrogen for gradually clarifying the solution, completing the reaction, spin-drying the solvent, adding a little dichloromethane for redissolution in normal hexane, layering, and taking down the oily substance of the lower layer to obtain 0.24 g of Alk-NCA as brown oily substance.
(2) The Eth-NCA is prepared by weighing 1.0g of L-glutamic acid, 10.3mL of 3-butene-1-ol and 1mL of trimethylchlorosilane in a 50mL eggplant bottle, heating at 60 ℃ for reaction for 5 hours, gradually clarifying the solution, stopping heating after the reaction is finished, spin-drying the solvent, and centrifuging by a small amount of repeated precipitation in diethyl ether to obtain 2a 1.34g of intermediate as a white solid. Weighing intermediate 2a.1 g and triphosgene 0.7g, placing the mixture into a 100mL eggplant bottle, adding ultra-dry tetrahydrofuran 25mL, reacting at room temperature, gradually changing the solution from turbidity into light yellow clear solution (6 h), ending the reaction, spin-drying the solvent, adding a little methylene chloride for re-dissolution, layering in normal hexane, and taking down oily substance to obtain Eth-NCA0.71 g which is light yellow oily;
(3) the preparation method of the tBA-NCA comprises the steps of weighing 0.54g of gamma-tertiary butyl-L-glutamic acid and 0.72g of triphosgene in a 100mL eggplant bottle, adding 16mL of ultra-dry tetrahydrofuran, stirring at 25 ℃ for 2h under a nitrogen atmosphere, gradually clarifying the solution, ending the reaction, decompressing and spin-drying the solvent, slowly dripping 15mL of ultra-dry N-hexane (15 min) into the eggplant bottle, stirring for 1h under the N 2 atmosphere until no solid is precipitated, filtering, washing with a small amount of ultra-dry N-hexane for many times, and drying overnight in a vacuum drying oven to obtain 0.53 g of tBA-NCA which is white solid.
The specific experimental procedure or conditions are not noted in the examples, and the operation or conditions of the conventional experimental procedure described in the literature in the field are sufficient. The raw materials or instruments used are all conventional products obtained by commercial use, including but not limited to those used in the examples of the present application.
Example 1
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Co, and has a hydrodynamic diameter of 10nm;
the preparation method of the protein-like particles comprises the following steps:
Dissolving polyethylene glycol methyl ether amine (100 mg, with a structural formula of MeO-PEO 5k-NH2) with a molecular weight of 5000 in 2mLDMF to obtain polyethylene glycol methyl ether amine solution, dissolving Alk-NCA monomer (225 mg) with 2mL DMF, adding the solution into the polyethylene glycol methyl ether amine solution, stirring at room temperature (25 ℃) for 3 days, and carrying out ring-opening polymerization to obtain a first block polymer, namely PEO 5k-Alk30;
Eth-NCA monomer (227 mg) was dissolved in 2mL of DMF, added to the first block polymer, stirred at room temperature (25 ℃) for 1 day to give a mixture, and finally tBA-NCA monomer (252 mg) was dissolved in 2mL of DMF, added to the mixture, stirred at room temperature (25 ℃) for 1 day, polymerization was completed, solvent in the product of the polymerization was spin-dried, redissolved with methylene chloride, precipitated in diethyl ether, centrifuged at 5000rpm for 30min, and the supernatant was discarded, and repeated 3 times to give a sequential polymer PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (i.e., second block polymer) as a brown solid.
The PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 5mL of DMP (dimethyl phthalate) to obtain a mixture, 6.1mg of Co 2(CO)8 was dissolved with 5mL of DMP to obtain Co 2(CO)8 solution, the mixture and Co 2(CO)8 solution were simultaneously and slowly dropped into 40mL of DMP, and stirred for 2 days to perform a complex crosslinking reaction (FIG. 2A) to obtain single-chain nanoparticles;
Subsequently, 15mL of single-stranded nanoparticles were taken under nitrogen flow, heated at 180℃for 12 hours, subjected to thermal decomposition reaction to obtain thermally decomposed nanoparticles, and then mixed with 35. Mu.L of trifluoroacetic acid, subjected to deprotection reaction at 25℃for 48 hours, and t-butyl was removed to obtain protein-like particles (B in FIG. 2).
FIG. 2 is a TEM image of single-stranded nanoparticles and protein-like particles prepared in example 1 of the present invention, wherein A in FIG. 2 is a single-stranded nanoparticle and B in FIG. 2 is a protein-like particle. As shown in FIG. 2, the nano particles are uniformly dispersed, the particle size is stable within 10-20 nm, and no large particles or agglomeration phenomenon is observed, so that the nano particles have good dispersibility and uniformity.
FIG. 3 is a block diagram of PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer and protein-mimetic particles prepared according to example 1 of the present invention, wherein A in FIG. 3 is PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer and B in FIG. 3 is protein-mimetic particles.
FIG. 4 is a synthetic route of the protein-like particles prepared in example 1 of the present invention.
FIG. 5 is a hydrogen spectrum of the first block polymer PEO 5k-Alk30 prepared in example 1 of the present invention. From fig. 5, the average degree of polymerization of AlkNCA monomers was calculated to be 30 by nuclear magnetic hydrogen spectrometry, m.w. =10430 g/mol for peo 5k-Alk30.
FIG. 6 is a hydrogen profile of PEO 5k-Alk30-Eth55-tBA52 prepared in example 1 of the present invention. From fig. 6, the average degrees of polymerization of EthNCA and tBANCA monomers were calculated to be 55 and 52, respectively, by nuclear magnetic hydrogen spectrometry, m.w. =30115 g/mol for peo 5k-Alk30-Eth55-tBA52.
GPC graphs of MeO-PEO 5k-NH2、PEO5k-Alk30 and PEO 5k-Alk30-Eth55-tBA52 were examined and shown in FIG. 7, and the distribution and polydispersity of MeO-PEO 5k-NH2 (as initiator), two-block polymer PEO 5k-Alk30 and four-block polymer PEO 5k-Alk30-Eth55-tBA52 were determined by GPC, and the results are shown in Table 3 below, to further verify the molecular weight of the polymers. As can be seen from FIG. 7 and Table 3, the higher the molecular weight, the earlier the peak was generated during GPC detection, the GPC peak elution time corresponded to the molecular weight of each polymer, the molecular weight of PEO 5k-Alk30-Eth55-tBA52 was the largest, the peak generation time was the earliest (15.63 min), and the MeO-PEO 5k-NH2 initiator molecular weight was the smallest and the peak generation time was the latest (15.83 min). The PDI of each block polymer is below 1.2, which indicates that the synthesis process of the polymer is well controlled, the molecular weight distribution of the polymer is narrower, the difference among the molecular weights of the polymers is small, and the polymer has good dispersibility and uniformity.
TABLE 3 molecular weight and polydispersity information for MeO-PEO 5k-NH2、PEO5k-Alk30 and PEO 5k-Alk30-Eth55-tBA52
| Polymer | Mn(NMR) | Peak time (min) | PDI(GPC) |
| MeO-PEO5k-NH2 | 5000 | 15.83 | 1.1 |
| PEO5k-Alk30 | 10430 | 15.67 | 1.2 |
| PEO5k-Alk30-Eth55-tBA52 | 30115 | 15.63 | 1.2 |
Example 2
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Co, and has a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 10mL of o-DCB, swelled, dissolved after 5mL of DMF was added to obtain a mixture, 12.2mg of Co 2(CO)8 was dissolved with 10mL of o-DCB to obtain Co 2(CO)8 solution, the mixture and Co 2(CO)8 solution were simultaneously and slowly dropped into 5mL of DMF, stirred for 2 days, subjected to complexation and crosslinking reaction, then heated at 180℃for 12 hours, subjected to thermal decomposition reaction to obtain thermally decomposed nanoparticles, and then mixed with trifluoroacetic acid, subjected to deprotection reaction at 25℃for 4 hours to remove tert-butyl groups to obtain a protein-like particle.
Example 3
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Co, and has a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 5mL THF, 6.1mg Co 2(CO)8 was slowly dropped into 5mL THF while stirring for 2 days to carry out complexation crosslinking reaction, then, heating at 180℃for 12 hours to carry out thermal decomposition reaction to obtain a thermally decomposed nanoparticle, mixing the thermally decomposed nanoparticle with trifluoroacetic acid, carrying out deprotection reaction at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 4
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Co, and has a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 10mL DMA, 12.2mg Co 2(CO)8 was slowly dropped into 5mL DMA while stirring for 2 days, and then, heating at 180℃for 12 hours, and then, thermal decomposition reaction was performed to obtain a thermally decomposed nanoparticle, and then, the thermally decomposed nanoparticle was mixed with trifluoroacetic acid, and a deprotection reaction was performed at 25℃for 48 hours to remove t-butyl groups, to obtain a protein-like particle.
Example 5
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Fe, and the hydrodynamic diameter is 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 90mL o-DCB, 13.9mg Fe (CO) 5 was dissolved with 10mL o-DCB, the polymer solution was slowly dropped into Fe (CO) 5 solution, stirred for 2 days, and subjected to complex crosslinking reaction, followed by heating at 180℃for 12 hours to give thermally decomposed nanoparticles, and then mixing the thermally decomposed nanoparticles with trifluoroacetic acid, and then carrying out deprotection reaction at 25℃for 48 hours to remove t-butyl groups, to give a protein-like particle.
Example 6
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Fe, and the hydrodynamic diameter is 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 45mLDMP, 7.0mg Fe (CO) 5 was dissolved with 5mL DMP, the polymer solution was slowly dropped into the Fe (CO) 5 solution, stirred for 2 days, and subjected to complex crosslinking reaction, followed by heating at 180℃for 12 hours to obtain thermally decomposed nanoparticles, and then mixing the thermally decomposed nanoparticles with trifluoroacetic acid, and carrying out deprotection reaction at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 7
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Co, and has a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 5mL of DCM, 6.1mg of Co 2(CO)8 was slowly dropped into 40mL of DCM while stirring for 2 days, and complexation crosslinking reaction was performed, then 15mL of the reaction solution was taken under nitrogen flow, the solvent was replaced with dimethyl phthalate at 180℃and heated at 180℃for 12 hours to obtain thermally decomposed nanoparticles, and then the thermally decomposed nanoparticles were mixed with 35. Mu.L of trifluoroacetic acid, deprotected at 25℃for 48 hours to remove tert-butyl groups to obtain the protein-like particle.
Example 8
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Fe, and the hydrodynamic diameter is 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 5mLDMP, 6.5mg Fe 2(CO)9 was slowly dropped into 40mL DMP while stirring for 2 days, and then 15mL of the reaction solution was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 9
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroidic nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Mn, and the hydrodynamic diameter is 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (50 mg) was dissolved in 5mLDMP, 6.9mg Mn 2(CO)10 was dissolved with 5mL DMP while being slowly dropped into 40mL DMP, and stirred for 2 days to carry out a complex crosslinking reaction, then 15mL of the reaction solution was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 10
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid-like nano particles, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Cr with a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 5mL of DMP, 15.7mg of Cr (CO) 6 was simultaneously dissolved in 5mL of DMP and slowly dropped into 40mL of DMP while stirring for 2 days, and then a reaction solution of 15mL was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 11
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroid nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Mo, and the hydrodynamic diameter is 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 5mL of DMP, 18.8mg of Mo (CO) 6 was dissolved in 5mL of DMP while slowly dropping into 40mL of DMP, stirring was carried out for 2 days, and then 15mL of the reaction solution was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 12
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroidic nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of W with a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 5mL of DMP, 25.0mg of W (CO) 6 was simultaneously dissolved in 5mL of DMP and slowly dropped into 40. 40mLDMP while stirring for 2 days, and then 15mL of the reaction solution was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at 25℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
Example 13
The protein-like particle has a structure of A-B-C, wherein a chain segment A is a linear polymer chain with hydrophilicity, B is spherical or spheroidic nanoparticle, a chain segment C is a polyamino acid chain segment with a side chain containing double bonds and carboxyl groups, the number average molecular weight of the chain segment A is 5000, the number average molecular weight of the chain segment C is 19700, and the chain segment B consists of Ni and has a hydrodynamic diameter of 10nm;
A protein-like particle was prepared according to the method of example 1, except that PEO 5k-Alk30-Eth55-tBA52 tetrablock copolymer (100 mg) was dissolved in 5mL of DMP, 12.1mg of Ni (CO) 4 was simultaneously dissolved in 5mL of DMP and slowly dropped into 40mL of DMP while stirring for 2 days, and then a reaction solution of 15mL was heated at 180℃for 12 hours to obtain a thermally decomposed nanoparticle, and then the thermally decomposed nanoparticle was mixed with 35. Mu.L of trifluoroacetic acid, and deprotected at (25) ℃for 48 hours to remove t-butyl groups to obtain a protein-like particle.
The protein-like particle provided by the invention has an asymmetric structure and excellent mobility, multiple double bonds are arranged on the protein-like particle, the protein-like particle can be used as a nano carrier to be connected with a positive drug through click chemistry, the multi-carboxyl structure enables the protein-like particle to have good biocompatibility, can be connected with a targeting peptide through DCC coupling and has targeting property, and the nano particle has small size, can be discharged out of the body through kidneys in vivo, and has high safety to a human body and small toxic and side effects.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
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| CN118638410A (en) * | 2024-04-26 | 2024-09-13 | 首都医科大学 | Magnetic nanoparticles and preparation method and application thereof |
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2025
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