Nucleic acid delivery vector, preparation therefor, and use thereof

Abstract

Provided is a polycarbonate or a salt thereof. Further provided are a preparation method for the polycarbonate, a complex thereof with at least one nucleic acid, and a use thereof for delivering at least one nucleic acid.

Description

Nucleic acid delivery vectors, their preparation and application Technical Field

[0001] This invention relates to the fields of functional polymer nanomaterials and biomedical technology; specifically, it relates to a nucleic acid delivery carrier, its preparation method, and its use for delivering nucleic acids. Background Technology

[0002] Nucleic acid drugs have become one of the breakthrough means of preventing and treating diseases. However, due to their electrical charge, low stability, and large size, naked nucleic acids are susceptible to enzymatic degradation and have difficulty crossing biological barriers. Since nucleic acid drugs must be administered intracellularly or even within the cell nucleus to exert their effects, their clinical and industrial applications are highly dependent on safe and efficient delivery vectors.

[0003] Currently, common delivery vectors are mainly divided into viral vectors and non-viral vectors. Viral vectors were first developed for nucleic acid delivery due to their efficient cell infection capabilities, but their risks of insertional mutations, adverse immune responses, limited loading capacity, and complex preparation processes severely restrict their large-scale production and industrial application. Given the limitations of viral vectors, non-viral vectors have become a highly promising alternative for nucleic acid drug delivery. As the most extensively researched and clinically applied non-viral vector in recent years, lipid nanoparticles (LNPs) have achieved significant success in the delivery of siRNA drugs and mRNA vaccines. However, the stability and delivery efficiency of LNPs still need further improvement, and their inflammatory properties and potential biotoxicity have also raised widespread controversy and concerns. As another type of non-viral nucleic acid vector, polymer nanoparticles (PNPs) have seen slower research and application development. Due to numerous problems with existing PNP materials in terms of biocompatibility, delivery efficiency, and controllable large-scale production, and the fact that some polymers (such as PEI) can accumulate in vivo and cause long-term side effects, no PNP material has yet been found that promises to achieve clinical translation and industrial application of nucleic acid drugs.

[0004] There is an urgent need in this field for a novel delivery vector that is easy to mass-produce, has low biotoxicity, and can achieve efficient delivery of nucleic acids. Summary of the Invention

[0005] To address this, this paper proposes a novel, rapidly biodegradable polymer delivery carrier that overcomes the long-standing but unresolved problems of high toxicity and low efficiency in existing polymer carriers. This provides a safe, effective, and reliable means for nucleic acid delivery and is suitable for large-scale production and industrial applications.

[0006] On the one hand, this document provides a polycarbonate or a salt thereof, said polycarbonate having repeating units of Formula I:

[0007] in,

[0008] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group, with the constraint that at least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0009] L is a key, C 1-16 Hydroxyl or carbonate bond;

[0010] R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0011] a1 and a4 are each independent integers from 1 to 16;

[0012] a2 and a3 are each independent integers from 0 to 16; and

[0013] b1 to b4 are each an independent integer from 0 to 2.

[0014] On the other hand, this article provides a method for preparing polycarbonate, comprising a polymerization reaction using a monomer having Formula II and a monomer having Formula III:

[0015] In Equations II and III,

[0016] X 1 and X 2 Each independently is: a key (such as a single key) or

[0017] Y 1 and Y 2 Each independently is: a key (such as a single key) or

[0018] X, Y, R 1 L, a1 to a4, b1 to b4 are each defined as above with respect to equation I;

[0019] The restrictions are:

[0020] (1) Equation II and Equation III together have at least one X and at least one Y, and at least one X and at least one Y do not come from Equation II or Equation III at the same time;

[0021] (2) At least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0022] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0023] On the other hand, this document provides a complex of the polycarbonate or its salt with at least one nucleic acid.

[0024] On the other hand, this article provides a method for preparing polycarbonate nucleic acid complexes, which includes:

[0025] - Provides polycarbonate or a salt thereof, and at least one nucleic acid; and

[0026] - Mix the polycarbonate or its salt with the at least one nucleic acid.

[0027] On the other hand, this document provides a composition comprising the polycarbonate or a salt thereof, or the complex thereof.

[0028] On the other hand, this article provides a product comprising:

[0029] -The complex;

[0030] -The composition; or

[0031] - The polycarbonate or its salt and at least one nucleic acid.

[0032] On the other hand, this article provides a method for delivering at least one nucleic acid to a cell or object, the method comprising:

[0033] - Provide the complex, the complex being formed by combining a polycarbonate or a salt thereof with the at least one nucleic acid; and

[0034] - To bring the cell or the object into contact with the complex.

[0035] On the other hand, this article provides a method for expressing at least one nucleic acid in a cell or object, the method comprising:

[0036] - Provide the complex, the complex being formed by combining a polycarbonate or a salt thereof with the at least one nucleic acid; and

[0037] - To bring the cell or the object into contact with the complex.

[0038] On the other hand, this document provides the use of the polycarbonate or its salts as nucleic acid carriers for delivering the nucleic acid to cells or objects.

[0039] On the other hand, this document provides the use of the polycarbonate or its salts, complexes or compositions for delivering at least one nucleic acid to cells or objects.

[0040] On the other hand, this document provides the use of the polycarbonate or its salts, complexes or compositions for expressing at least one nucleic acid in cells or objects.

[0041] On the other hand, this document provides the use of the polycarbonate or its salts, complexes or compositions in the preparation of products for delivering at least one nucleic acid to cells or objects.

[0042] On the other hand, this document provides the use of the polycarbonate or its salts, complexes or compositions in the preparation of products for enabling cells or objects to express at least one nucleic acid. Attached Figure Description

[0043] The present invention will be further described below with reference to the accompanying drawings, which are shown only for illustrating embodiments of the present invention and are not intended to limit the scope of the present invention.

[0044] Figure 1 shows a schematic diagram of the formation of polycarbonate-nucleic acid complexes by polycarbonate and nucleic acids according to this paper.

[0045] Figure 2 shows the results of cell safety tests using the exemplary polycarbonate / Luc-mRNA complex according to this article.

[0046] Figure 3 shows the results of cell safety tests using the exemplary polycarbonate / Luc-mRNA complex according to this article.

[0047] Figure 4 shows the results of cell safety tests using the exemplary polycarbonate / pCMV-luc complex according to this article.

[0048] Figure 5 shows the results of cell transfection using the exemplary polycarbonate / Luc-mRNA complex according to this article.

[0049] Figure 6 shows the results of cell transfection using the exemplary polycarbonate / Luc-mRNA complex according to this article.

[0050] Figure 7 shows the results of cell transfection using the exemplary polycarbonate / pCMV-luc complex according to this article.

[0051] Figure 8 shows polymers 791 and 250. 13 C10 NMR spectrum (150MHz, D2O).

[0052] Figure 9 shows polymers 791 and 792. 13 C10 NMR spectrum (150MHz, CDCl3).

[0053] Figure 10 shows polymers 790 and 251. 13 C10 NMR spectrum (150MHz, D2O).

[0054] Figure 11 shows polymers 790 and 792. 13 C10 NMR spectrum (150MHz, DMSO-d6).

[0055] Figure 12 shows bioluminescent images (A) and bioluminescent quantification (B) of BALB / c mice 24 hours after intramuscular injection of polymer 791 / Luc-mRNA complex, polymer 790 / Luc-mRNA complex, and PEI / Luc-mRNA complex (10 μg mRNA) (n = 3, ****p < 0.0001).

[0056] Figure 13 shows bioluminescence images (A) and bioluminescence quantification (B) of BALB / c mice at different time points after intramuscular injection of polymer 791 / Luc-mRNA complex and polymer 790 / Luc mRNA complex (10 μg mRNA) (n = 3, *p < 0.05).

[0057] Figure 14 shows bioluminescence images (A) and bioluminescence quantification (B) of BALB / c mice at different time points after intratumoral injection of polymer 791 / Luc-mRNA complex, polymer 790 / Luc-mRNA complex, and LNPs / Luc-mRNA complex (10 μg mRNA) (n = 3, *p < 0.05, **p < 0.01, ***p < 0.01).

[0058] Figure 15 shows the CDS nucleotide sequence (SEQ ID No.:1) of the Luc-mRNA used to prepare the polycarbonate / Luc-mRNA complex.

[0059] Figure 16 shows the full-length nucleotide sequence of Luc-mRNA used to prepare the polycarbonate / Luc-mRNA complex (SEQ ID No.:2). Detailed Implementation

[0060] Nucleic acid molecules, as carriers and transmitters of genetic information, are among the most crucial biomolecules within cells. DNA is responsible for storing the genetic code and guiding cellular activities, while RNA participates in various biological processes, including acting as a genetic messenger (mRNA), participating in protein synthesis (tRNA and rRNA), and regulating gene expression (small RNA). In modern medicine, by delivering specific nucleic acid drugs to target sites, it is possible to express antigen proteins to stimulate an immune response, or to regulate or correct genetic information that leads to adverse consequences, thereby preventing or treating various diseases or conditions, including inflammatory conditions, cancer, and genetic diseases.

[0061] However, despite the enormous market potential of nucleic acid drugs, their practical application faces numerous obstacles. Nucleic acid molecules are highly susceptible to degradation before reaching their target location and are generally unable to effectively cross biological barriers (such as cell membranes) to enter the cell interior. This is because their large molecular weight and electrical charge limit their ability to pass through hydrophobic cell membranes.

[0062] Taking messenger RNA (mRNA) as an example, as a transient, encoding genetic information carrier transcribed from DNA strands and carrying information about protein synthesis, mRNA can guide the translation and expression of functional proteins in cells. mRNA-based therapies, by delivering mRNA encoding different proteins into the body for translation and expression, can produce almost any functional protein, thus holding promise for the treatment of various (refractory) diseases, including infectious diseases, metabolic genetic disorders, cancer, and cardiovascular diseases. Recently, two mRNA vaccines (mRNA-1273 and BNT162b2) produced by Moderna and Pfizer-BioNTech have been successfully used to prevent COVID-19, highlighting the enormous potential of mRNA technology to revolutionize life sciences and medical research. In addition to COVID-19 vaccines, several other mRNA vaccines targeting other infectious diseases (such as respiratory syncytial virus, seasonal influenza, and rabies virus), cancers (such as melanoma), and mRNA drugs for protein therapy and gene editing are currently in various stages of clinical trials. With breakthroughs in nucleic acid chemistry and RNA biology technologies, such as the modification of the untranslated region (UTR) of traditional linear mRNA and the emergence of circular mRNA, the most concerning issues of immunogenicity and protein expression efficacy in mRNA have been largely resolved. Therefore, the main challenge in the clinical translation of mRNA drug therapeutics lies in the development of mRNA delivery vectors.

[0063] However, mRNA is a large polyanionic compound that does not easily cross nonpolar cell membranes and tissue barriers. Furthermore, it is readily destroyed and inactivated by nucleases. Therefore, a delivery vector is needed to protect the mRNA and allow for endosome escape in order to deliver fully functional mRNA into the cytoplasm and exert its therapeutic effect. Due to poor degradation performance, existing mRNA delivery vectors generally suffer from toxicity and delayed nucleic acid release (low transfection efficiency). Specifically, if the delivery vector is non-degradable or its degradation products are highly toxic, safety issues arise; if the vector degrades too slowly, the mRNA cannot be released rapidly, limiting transfection efficiency.

[0064] Therefore, the development of safe and effective nucleic acid delivery vectors has become the biggest bottleneck limiting the potential of current nucleic acid drugs. Successfully developing such vectors would not only significantly improve delivery efficiency, thereby enhancing the efficacy of nucleic acids, but also enable their widespread application and promotion across multiple fields due to their sufficiently safe delivery process. This breakthrough will greatly promote the development of personalized and precision medicine, drive the transformation of the pharmaceutical industry, and significantly improve patients' treatment outcomes and quality of life.

[0065] Through long-term and extensive exploration and research, the inventors have pioneered the development of a novel carbonate polymer that can serve as a polymer nanocarrier for nucleic acid delivery. This polymer nanocarrier is stable in vitro and rapidly and completely self-degrades upon entering experimental or pharmacological target sites (such as cells), with degradation products consisting of highly safe small molecules (such as CO2 and thiols). Due to its unique physicochemical properties, this material can form complexes with a wide range of nucleic acids of varying molecular weights, lengths, and types, effectively protecting them from enzymatic degradation, enhancing their stability, and promoting transcellular transport, significantly improving the delivery efficiency of target nucleic acid molecules. Furthermore, the nanomaterial's preparation process is simple and environmentally friendly, suitable for large-scale production to meet the needs of a wide range of experimental and industrial applications, providing a completely new option for nucleic acid delivery.

[0066] definition

[0067] To facilitate a better understanding of this disclosure, some key terms are first defined. Unless otherwise expressly stated, all terms listed below should be understood based on the appended definitions. Unless otherwise stated, the meanings of other terms herein are consistent with the common understanding of those skilled in the art.

[0068] In this document, the word "one" or its combination with various quantifiers includes both singular and plural meanings. When multiple numerical values, ranges, or combinations thereof are given for the same parameter or variable, it is equivalent to specifically revealing these numerical values, range endpoints, and the ranges formed by any combination of them. In this document, any numerical value, regardless of whether it contains modifiers such as "approximately," encompasses an approximate range that can be understood by those skilled in the art, such as ±20%, ±10%, ±5%, etc.

[0069] In this document, each “implementation” equally refers to and encompasses all implementations of the methods and systems described herein. One or more technical features in any implementation can be freely combined with one or more technical features in any one or more other implementations, and the resulting implementation is also part of the disclosure herein.

[0070] Unless otherwise specified, all percentages and ratios used herein are by weight. Unless otherwise specified, all temperatures are expressed in degrees Celsius (°C). Room temperature or ambient temperature refers to temperatures between 20°C and 28°C (e.g., 25°C). Unless otherwise specified, all measurements are understood to be taken under ambient conditions, i.e., room temperature, approximately one atmosphere of pressure, and approximately 50% relative humidity.

[0071] All numerical ranges are narrower ranges that include the extreme values. The described upper and lower limits of the range can be combined to form additional ranges that are not explicitly described.

[0072] In this document, the term "polycarbonate" refers to a class of polymers in which at least two repeating units are linked by carbonate bonds -OC(O)O-. The terms "polycarbonate" and "carbonate polymer" are used interchangeably and have the same meaning herein. It should be understood that when referred to herein as "polycarbonate," it means polycarbonate and its salt forms. Salts of said polycarbonates are formed by salting with anions such as chloride, acetate, citrate, and tetrafluoroacetate.

[0073] In this article, the term "polymer" refers to a high molecular weight compound formed by repeating monomer units linked by covalent bonds. These repeating monomer units can be the same, forming a "homopolymer," or different, forming a "copolymer."

[0074] In this article, the term "repeating unit" refers to a polymeric unit that appears at least twice in the polymer backbone. Repeating units typically originate from monomers. The term "monomer" refers to a small molecule compound capable of linking together to form a long chain through polymerization. During polymerization, the multiple chemical bonds in monomer molecules break and reform, resulting in the repeated linking of monomeric units to form the polymer backbone.

[0075] In this article, the term "linear polymer" refers to a polymer with a linear molecular chain structure. The smaller side groups of linear polymers are not branched structures. The term "crosslinked polymer" refers to a polymer (e.g., obtained through a crosslinking reaction) that has a three-dimensional network structure.

[0076] In this article, the term "random copolymer" refers to a product obtained by random copolymerization, in which two or more monomer units are randomly distributed along the main chain, and no single monomer unit can form a separate long segment on the polymer chain. The term "block copolymer" refers to a copolymer in which two or more monomer units each form a long sequence of segments and are covalently bonded to each other.

[0077] In this article, the term "main chain" refers to the longest linear molecular chain that constitutes the polymer. All other molecular chains can be considered as side groups or side chains relative to the main chain. For example, the polymer main chain may consist primarily of carbon atoms linked by covalent bonds, and may be interspersed with atoms such as nitrogen, oxygen, and sulfur. Generally, terminal groups present at the ends of the polymer are not included in the main chain. When referring to "having" a group or atom "in the main chain," or a group or atom "located in the main chain," it means that the group or atom constitutes part of the polymer main chain (rather than a side group or end). It can be considered that a break at a position of a group or atom "located in the main chain" will result in the breakage of the polymer molecular chain.

[0078] In this article, the term "amino" refers to the functional group of an amine, and may include primary amino, secondary amino, tertiary amino, and quaternary amino groups.

[0079] In this article, the term "alkyl" refers to a saturated hydrocarbon group, which can be straight-chain or branched (including branched-chain) structures, and is generally a chain group without cyclic structures. The reference to "C"... 1-22 When "alkyl" is used, it refers to an alkyl group with 1-22 carbon atoms. Examples of such alkyl groups include methyl, ethyl, propyl (e.g., n-propyl), butyl (e.g., isopropyl, n-butyl, isobutyl, tert-butyl), pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, and their branched isomers.

[0080] In this article, the term "alkenyl" refers to an unsaturated hydrocarbon group containing at least one carbon-carbon double bond (C=C), which can be straight-chain or branched (including branched-chain) structures, and is generally a chain group without cyclic structures. The reference to "C..." 2-22 When "alkenyl" is used, it refers to an alkenyl group with 2-22 carbon atoms. Examples of such alkenyl groups include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, and 2-methylbut-2-enyl.

[0081] In this article, the term "alkynyl" refers to a hydrocarbon group containing at least one carbon-carbon triple bond (C≡C), which can be straight-chain or branched (including branched-chain) and is generally a chain group without cyclic structures. The reference to "C" is further elaborated. 2-22 When "alkynyl" is used, it refers to an alkynyl group with 2-22 carbon atoms. Examples of such alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, and 1-methylpent-2-alkynyl.

[0082] In this article, the terms "alkyl," "alkenyl," and "alkynyl" are collectively referred to as "hydrocarbon groups." The reference to "C" is also included. 1-22 When "hydrocarbon group" is used, it refers to a straight-chain or branched hydrocarbon group with 1-22 carbon atoms, namely alkane group, olefin group, or alkyne group. "Alkyl group" refers to a divalent hydrocarbon group, including alkylene group, alkenylene group, and alkyne group, which can be straight-chain or branched.

[0083] In this article, the term "cyclic hydrocarbon group" refers to a saturated or unsaturated cyclic hydrocarbon group, including "cycloalkyl," "cycloalkenyl," and "cycloynyl." The reference to "C" is also included. 1-20 When "cyclic hydrocarbon group" is used, it refers to a cyclic hydrocarbon group with 1-20 carbon atoms. Examples of such cyclic hydrocarbon groups can include monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclopentynyl, cyclohexynyl, and cycloheptynyl; bridged cyclic hydrocarbon groups, such as spirocyclic and norbornyl; and fused cyclic hydrocarbon groups, such as decahydronaphthyl and adamantyl.

[0084] In this document, the term "heterocyclic alkyl group" refers to an organic cyclic group comprising at least one (e.g., 1, 2, or 3) cyclic heteroatoms selected from the group consisting of N, O, S, Si, and P (e.g., selected from N, O, and S). Examples of such heterocyclic alkyl groups may include morpholino, piperazino, piperidino, furanyl, pyrroleo, thiopheno, pyranoyl, etc.

[0085] In this article, the term "heteroatom" refers to a non-carbon atom selected from oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0086] In this article, "aryl" refers to an aromatic cyclic group, that is, a cyclic structure with a planar or near-planar shape. Most aryl groups have a benzene ring structure. Aryl groups can be broadly classified into monocyclic aryl groups (e.g., phenyl), polycyclic aryl groups (e.g., biphenyl, diphenylmethyl, triphenylmethyl), and fused-ring aryl groups (e.g., naphthyl, anthracene, phenanthryl, pyrene, azulel, tetraphenyl).

[0087] In this document, the term "5- to 14-membered heteroaryl" refers to an aryl group having 5 to 14 (i.e., 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) ring skeleton atoms and including at least one (e.g., 1, 2, 3, or 4) ring heteroatoms selected from the group consisting of N, O, S, Si, and P (e.g., selected from N, O, and S). Such heteroaryl groups can be monocyclic or fused rings condensed with at least one benzene ring, and can be heteroaryl groups formed by linking at least one heteroaryl or aryl group to a heteroaryl group via one or more single bonds. Such heteroaryl groups can include monocyclic and fused ring heteroaryl groups. Examples of monocyclic heteroaryl groups may include furanyl, thiophene, pyrrole, imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, carbazole, triazinyl, tetraazinyl, triazolyl, tetraazolyl, furazinyl, pyridyl, pyrazinyl, pyrimidinyl, and pyridazinyl. Examples of fused-ring heteroaryl groups may include benzofuranyl, benzothiopheneyl, isobenzofuranyl, dibenzofuranyl, dibenzothiopheneyl, benzimidazolyl, benzothiazolyl, benzoisothiazolyl, benzoisooxazolyl, benzooxazolyl, isoindolyl, indolyl, benzoindolyl, indazole, benzothiadiazolyl, quinolinyl, isoquinolinyl, cenolinyl, quinazolinyl, quinoxalolinyl, benzoquinoxalolinyl, naphridinyl, carbazolyl, benzocarbazolyl, phenoxazinyl, phenthiazinyl, phenanthidyl, benzodioxacyclopentenyl, and dihydroacridinyl.

[0088] In this document, the term "3 to 10-membered heterocyclic alkyl" refers to an organic cyclic group having 3 to 10 (i.e., 3, 4, 5, 6, 7, 8, 9, 10, or a range formed with any two of the above values ​​as endpoints) cyclic skeletal atoms and including at least one (e.g., 1, 2, or 3) cyclic heteroatoms selected from the group consisting of N, O, S, Si, and P (e.g., selected from N, O, and S). Examples of such heterocyclic alkyl groups may include morpholinyl, piperazinyl, piperidinyl, furanyl, pyrroleyl, thiophenyl, pyranyl, etc. The heterocycle can be alicyclic (e.g., tetrahydrofuran) or aromatic (e.g., pyridine). Complex heterocyclic systems can be formed by the fusion of two or more simple rings (e.g., indole).

[0089] In this article, "C" x-y "This means that the corresponding group or part has x to y (including the terminal value) carbon atoms. For example, C..." 1-22 This represents the number of carbon atoms as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or within a range formed by any two of the above values ​​as endpoints. "C 3-20 “C” 1-16 “C” 1- 12 “C” 1-8 “C” 1-6“C” 1-4 “C” 3-14 “C” 6-14 "etc." have similar meanings.

[0090] In this article, the term "halogen" includes fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0091] In this article, the term "ester group" refers to a carboxylic acid ester group, i.e., a molecular chain containing -C(O)OR. x The structural group, wherein R x C 1-12 Alkyl groups, such as C 1-8 Alkyl, C 1-6 Alkyl or C 1-4 Alkyl groups. Examples of such ester groups may include methyl ester, ethyl ester, propyl ester, butyl ester, etc.

[0092] In this article, the term "amide group" refers to a group represented by any of the following chemical formulas: -CONH2, -CONH-, -CON<.

[0093] In this text, the term "substituted or unsubstituted" refers to the substitution of a hydrogen atom in a functional group by another atom or functional group (i.e., a substituent). For example, a substituted methyl group refers to a methyl group (-CH3) in which one or more hydrogen atoms are replaced by other atoms or functional groups (i.e., substituents). In this text, the substituents of a "substituted" group may be independently selected from: deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, ester, C... 1-22 Alkyl, Halogenated C 1-22 Alkyl, C 2-22 alkenyl, C 2-22 alkynyl group, C 1-22 Alkoxy, C 1-22 Alkylthio, C 1-22 Alkyl ester group, C 3-20 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-20 Aryl, C 6-20 aryloxy group, C 6-20 Arylthioyl, 5- to 14-membered heteroaryl, C 1-22 Alkyl C 3-20 cycloalkyl, C 1-22 Alkyl 3- to 10-membered heterocyclic alkyl, C 1-22 Alkyl C 6-20 Aryl, C 1-22 Alkylsilyl, C 6-20 Arylsilane, C 1-22 Alkyl C 6-20 Arylsilane, C 1-22 Alkylamino, C 6-20 arylamino, C 1-22 Alkyl C6-20 arylamino, C 1-22 alkyl carbonyl, C 1-22 alkoxycarbonyl, C 6-20 aryl carbonyl, C 6-20 arylboryl, C 1-22 Alkylboryl, C 1-22 Alkyl C 6-20 arylboryl and C 6-20 Aryl C 1-22 alkyl.

[0094] In this document, the term "redox-sensitive group" refers to a group whose molecular structure is responsive to one or both of oxidizing and reducing environments; that is, a group that readily undergoes a chemical reaction under oxidizing or reducing conditions. Exemplary oxidizing environments include highly reactive oxygen species (ROS) environments. Exemplary reducing environments include high glutathione (GSH) environments. In this document, the term "complex" means the product obtained by combining the polycarbonate described herein with at least one nucleic acid through non-covalent interactions (e.g., electrostatic interactions, ionic interactions, hydrogen bonds, or van der Waals forces). "Polycarbonate complex," "carbonate polymer complex," and "PC complex" are used interchangeably and have the same meaning herein.

[0095] In this article, the term "nucleotide," also known as "mononucleotide," refers to a compound composed of a base, a pentose sugar (deoxyribose or ribose), and a phosphate group linked together in a specific manner. It is the building block of nucleic acids (such as DNA and RNA). Nucleotides can polymerize through phosphodiester bonds to form dinucleotides, trinucleotides, and polymers such as oligonucleotides (less than 25 nucleotides) and polynucleotides (more than 25 nucleotides).

[0096] In this article, the term "nucleic acid" refers to a class of widely distributed biological macromolecules. Based on their composition, structure, and function, they can be mainly divided into ribonucleic acid (RNA) and deoxyribonucleic acid (DNA). RNA is composed of nucleotides linked by 3,5'-phosphodiester bonds, while DNA is composed of deoxynucleotides linked by 3',5'-phosphodiester bonds. In this article, the term "endogenous nucleic acid" refers to nucleic acid molecules that naturally exist within cells or organisms and are produced or inherited by the target cell or organism itself; the term "exogenous nucleic acid" refers to nucleic acid molecules that cannot be produced or inherited by the target cell or organism itself and originate from external sources.

[0097] In this document, "pharmaceutically acceptable" means a substance that, within reasonable medical judgment, is suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and is commensurate with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" includes pharmaceutically acceptable substances, compositions, or carriers suitable for administration to mammals of the polycarbonates or polycarbonate complexes described herein.

[0098] In this article, the term "unit dose" refers to the amount of substance expected to produce a single effect (e.g., prevention or treatment). Unit doses are typically prepared for a single use or for a single subject.

[0099] polycarbonate

[0100] On the one hand, this document provides a polycarbonate or a salt thereof, said polycarbonate having repeating units of Formula I:

[0101] in,

[0102] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group, with the constraint that at least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0103] L represents a key (such as a single key), C represents a key (such as a single key). 1-16 (For example, C) 1-12 C 1-10 C 1-8 C 1-6 C 1-4 Or C 1-3 ) Hydroxyl group (e.g., alkylene or alkenylene) or carbonate bond;

[0104] R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0105] a1 and a4 are each an integer from 1 to 16, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or an integer within a range formed by taking any two of the above values ​​as endpoints.

[0106] a2 and a3 are each independently an integer from 0 to 16, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or an integer within a range formed by taking any two of the above values ​​as endpoints; and

[0107] b1 to b4 are each an independent integer from 0 to 2, for example, 0, 1, or 2.

[0108] In Formula I, X and Y may each independently comprise: at least one nitrogen-containing group and / or at least one redox-sensitive group, provided that at least one of X and Y comprises at least one protonable nitrogen-containing group and X is different from Y. The nitrogen-containing group may be a protonable nitrogen-containing group or other nitrogen-containing groups, for example, as described herein. In some embodiments, X and Y each comprise at least one (e.g., at least two, three, or more) nitrogen-containing group. In some embodiments, X and Y each comprise at least one (e.g., at least two, three, or more) redox-sensitive group. In some embodiments, one of X and Y comprises at least one (e.g., at least two, three, or more) nitrogen-containing group, and the other comprises at least one (e.g., at least two, three, or more) redox-sensitive group. In some embodiments, one or both of X and Y comprises a carbonate group on the main chain. In some embodiments, one or both of X and Y does not comprise a carbonate group on the main chain. In some embodiments, one or both of X and Y does not comprise a carbonate group.

[0109] The polycarbonate described herein has one or more nitrogen-containing groups in its repeating unit (e.g., a repeating unit of Formula I). ​​The one or more nitrogen-containing groups may be located in the main chain and / or side groups of the polycarbonate. Preferably, at least one nitrogen-containing group is located in the main chain of the polycarbonate. For example, at least one (e.g., at least 1 or 2) nitrogen atoms of the at least one nitrogen-containing group are located in the main chain of the polycarbonate. In some embodiments, the number of nitrogen-containing groups or nitrogen atoms located in the main chain of the polycarbonate may be greater than the number of nitrogen-containing groups or nitrogen atoms located in the side groups. In some embodiments, the repeating unit of Formula I contains no more than 5, 4, 3, 2, or 1 nitrogen-containing groups or nitrogen atoms in the side groups, or none at all. In some embodiments, some or all of the nitrogen-containing groups or nitrogen atoms in the repeating unit of Formula I are located in the main chain of the polycarbonate. In some embodiments, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or 100% of the nitrogen-containing groups or nitrogen atoms in the repeating unit of Formula I are located in the main chain of the polycarbonate rather than in the side groups.

[0110] Advantageously, the polycarbonate is protonable. Specifically, the polycarbonate is protonable by incorporating at least one protonable nitrogen-containing group into its repeating unit (e.g., a repeating unit of Formula I). ​​In some embodiments, the at least one protonable nitrogen-containing group has at least one protonable nitrogen atom. In some embodiments, the protonable atom of the protonable nitrogen-containing group is nitrogen. In some embodiments, for example in a repeating unit of Formula I, at least one protonable nitrogen-containing group or at least one protonable nitrogen atom is located in the main chain of the polycarbonate.

[0111] In some embodiments, the protonable nitrogen-containing group is a divalent group.

[0112] In some exemplary embodiments, the protonable nitrogen-containing group may include groups selected from the group consisting of:

[0113] -A group having a protonable nitrogen atom located in the main chain and no side groups;

[0114] - A group having a protonable nitrogen atom located in the main chain and a hydrocarbon group (e.g., a saturated or unsaturated hydrocarbon group) attached to its side;

[0115] - A group having a protonable nitrogen atom located in the main chain and a side group containing a heteroatom (e.g., one or more of O, N, S, P, Si).

[0116] The protonatable groups on the polymer can absorb protons and acquire a positive charge, thereby imparting or increasing the positive charge of the polymer. Without being bound by any particular theory, it has been found that this change can advantageously affect the physicochemical properties of the polymer, helping it to effectively load nucleic acids.

[0117] In some embodiments, the protonable nitrogen-containing groups contained in the polycarbonate may have groups selected from the group consisting of amino groups, nitrogen-containing cyclic groups, and any combination thereof.

[0118] In cases where at least one protonable group comprises at least one amino group, the amino group may be selected from the group consisting of secondary amino groups, tertiary amino groups, quaternary amino groups, and combinations thereof.

[0119] In some embodiments, the amino group may be a monovalent amino group (having the structure -N(R)). 3 )2 or -N(R 3 )3 + ) or divalent amino (with the structure -N(R) 3 - or -N(R) 3 )2 + -), where R 3 Selected from hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, amide, urethane, urea, substituted or unsubstituted ester groups, substituted or unsubstituted C groups. 1-22 Alkyl, substituted or unsubstituted C 2-22 alkenyl, substituted or unsubstituted C 2-22 Alkyne group, substituted or unsubstituted C 1-22 Alkoxy, substituted or unsubstituted C 1-22 Alkylamino, substituted or unsubstituted C 1-22 Alkyl ester group, substituted or unsubstituted C 3-20 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl, substituted or unsubstituted 5- to 14-membered heteroaryl, substituted or unsubstituted C 1-22 Alkyl 3- to 10-membered heterocyclic alkyl groups, and substituted or unsubstituted C-membered alkyl groups. 1-22 Alkyl 5 to 14-membered heteroaryl groups.

[0120] In various specific implementations, R 3 Non-limiting examples may include:

[0121] in,

[0122] n1 and n2 are each an integer from 0 to 21 each time they appear, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or an integer within the range formed by taking any two of the above values ​​as endpoints;

[0123] Each occurrence of n3 is an integer from 2 to 21, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or an integer within the range formed by any two of the above values ​​as endpoints;

[0124] n4 and n5 are each an integer from 0 to 8 when they appear, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, or an integer within the range formed by taking any two of the above values ​​as endpoints;

[0125] Each occurrence of n6 is an integer from 0 to 8, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, or an integer within the range formed by any two of the above values ​​as endpoints;

[0126] Each occurrence of n7 is an integer from 1 to 8, for example, 1, 2, 3, 4, 5, 6, 7, 8, or an integer within the range formed by any two of the above values ​​as endpoints;

[0127] Among them, wavy lines Indicates the bonding position with adjacent atoms; and

[0128] Optionally, the above-mentioned groups may be substituted by one or more substituents selected from the group consisting of: deuterium, tritium, hydroxyl, carboxyl, nitro, amino, halogen, cyano, ester, C 1-12 Alkyl, C 2-12 alkenyl, C 2-12 alkynyl group, C 1-12 Alkoxy, C 1-12 Alkylamino, C 1- 12 Alkyl ester group, C 3-14 cycloalkyl, C 6-14 Aryl, 3- to 10-membered heterocyclic alkyl, 5- to 14-membered heteroaryl, C 1-12 Alkyl 5- to 14-membered heteroaryl, C 1-12 Alkyl groups of 3 to 10-membered heterocyclic alkyl groups, and any combination thereof.

[0129] In some embodiments, the polycarbonate contains an amino group in its main chain, or more specifically, one or more of secondary, tertiary, and quaternary amino groups, or more specifically, nitrogen atoms of one or more of secondary, tertiary, and quaternary amino groups. In some embodiments, the polycarbonate (e.g., in its main chain) may not contain a secondary amino group as the sole protonable nitrogen-containing group. In some embodiments, the polycarbonate (e.g., in its main chain) may not contain a secondary amino group as the sole protonable nitrogen-containing group. In some embodiments, the polycarbonate (e.g., in its main chain) may not contain any amino group other than a tertiary amino group as the sole protonable nitrogen-containing group. In some embodiments, the polycarbonate (e.g., in its main chain) may not contain any amino group other than a tertiary amino group as the protonable nitrogen-containing group.

[0130] In cases where at least one protonable group comprises at least one nitrogen-containing cyclic group, the nitrogen-containing cyclic group may be selected from: 3 to 10 saturated or unsaturated aliphatic or aromatic heterocyclic groups having at least one cyclic nitrogen atom, 5 to 20 fused 3 to 10 saturated or unsaturated aliphatic or aromatic heterocyclic groups having at least one cyclic nitrogen atom to each other or to an aliphatic or aromatic carbocyclic ring, and combinations thereof. Examples of such nitrogen-containing cyclic groups include, but are not limited to: (a)azacyclopropane, (a)azacyclobutane, (a)pyrroleyl, (a)thiazolyl, (a)oxazolyl, (a)pyrazolyl, (a)imidazolyl, (a)triazolyl, (a)piperidinyl, (a)morpholinyl, (a)thiomorpholinyl, (a)piperazinyl, (a)hexahydropyrimidinyl, (a)triazinyl, (a)azacycloheptane, (a)oxazacycloheptane, (a)homoperazinyl, (a)triazacycloheptane, (a)azacyclopropenyl, (a)azacyclobutenyl, (a)pyrroleyl, (a)pyrazolyl, (a)imidazolyl, (a)triazole (Ide)pyridyl, (ide)dihydropyridyl, (ide)tetrahydropyridyl, (ide)pyrazinyl, (ide)pyrimidinyl, (ide)pyridazinyl, (ide)acogenyl, (ide)diazazolyl, (ide)trizazolyl, (ide)thiazolyl, (ide)thiazolinyl, (ide)thiazinyl, (ide)oxazinyl, (ide)azacyclooctyl, (ide)oxazacyclooctyl, (ide)azacyclononyl, (ide)oxazacyclononyl, (ide)azacyclodecyl, (ide)oxazacyclodecyl, (ide)benzothiazolyl, (ide)benzothiazinyl, and 3 to 20-membered heterocyclic groups formed by their fusion with each other or with aliphatic or aromatic carbocyclic rings, and combinations thereof.

[0131] In some embodiments, the nitrogen-containing cyclic group may have the structure of formula Ia:

[0132] in,

[0133] Y 1 It is N or C; and

[0134] Ring R A It is a 3- to 20-membered aliphatic heterocycle having 1 to 6 heteroatoms selected from N, O, S and combinations thereof;

[0135] Optionally, where ring R A One or more ring carbon atoms and / or one or more ring nitrogen atoms are each independently bounded by one or two R atoms. 2 Replace; where R 2 Each group is independently selected from the following groups when it appears: deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, amide, urethane, urea, substituted or unsubstituted ester, substituted or unsubstituted C. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups, and substituted or unsubstituted 5- to 14-membered heteroaryl groups; and, two adjacent R groups 2 They can form rings together with the ring atoms they are connected to (e.g., forming monocyclic or bicyclic rings);

[0136] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0137] In some implementations, ring R A It can be a 3- to 20-membered aliphatic heterocycle (e.g., a 3- to 10-membered aliphatic heterocycle, such as a 3- to 7-membered aliphatic heterocycle) having 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, S, and combinations thereof.

[0138] In some embodiments, the nitrogen-containing cyclic group may have the structure of formula Ib:

[0139] in,

[0140] Y 2 and Y 4 Each is independently N or C, and Y 2 and Y 4 At least one of them is N;

[0141] Y 3 Selected from: bonds (such as single bonds), C, N, O, and S;

[0142] Y 5 Selected from: bonds (such as single bonds), C, N, O, and S;

[0143] Each occurrence of n is an integer between 0 and 2.

[0144] Optionally, where ring R B One or more ring carbon atoms and / or one or more ring nitrogen atoms are each independently bounded by one or two R atoms. 2 Replace; where each R 2 Each is defined independently according to the above definition of equation Ia;

[0145] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0146] In some implementations, ring R B It can be a 3- to 20-membered aliphatic heterocycle (e.g., a 3- to 10-membered aliphatic heterocycle, such as a 3- to 7-membered aliphatic heterocycle) having 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, S, and combinations thereof.

[0147] In some exemplary embodiments, the nitrogen-containing cyclic group may include the following groups:

[0148] Optionally, wherein one or more cyclic carbon atoms and / or one or more cyclic nitrogen atoms of the aforementioned nitrogen-containing cyclic group are each independently bound by one or two R groups. 2 Replace; where each R 2 Each is defined independently according to the above definition of equation Ia;

[0149] Optionally, one or more (e.g., 1, 2 or more) of the nitrogen-containing cyclic carbon atoms are each independently replaced by a heteroatom selected from N, O and S;

[0150] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0151] In some embodiments, the polycarbonate includes nitrogen-containing cyclic groups in its main chain. In some embodiments, the nitrogen-containing cyclic groups are linked to the main chain via one or two cyclic nitrogen atoms. In some embodiments, the polycarbonate (e.g., in the main chain) may not contain any nitrogen-containing groups other than the nitrogen-containing cyclic groups as the only protonable nitrogen-containing groups. In some embodiments, the polycarbonate (e.g., in the main chain) may not contain any nitrogen-containing groups other than the nitrogen-containing cyclic groups as protonable nitrogen-containing groups.

[0152] The protonable nitrogen-containing group may be unsubstituted or substituted with one or more substituents. Exemplary substituents may include: deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, amide, urethane, urea, substituted or unsubstituted ester groups, substituted or unsubstituted C groups. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl; and, the two adjacent substituents may form a ring together with the ring atom to which they are attached (e.g., forming a monocyclic or bicyclic ring).

[0153] In some embodiments, in addition to the protonable nitrogen-containing groups described above, the polycarbonate may also contain other nitrogen-containing groups in its repeating units (e.g., repeating units of Formula I), such as, but not limited to, nitrile, isonitrile, azide, hydrazone, nitro, nitroso, azaaryl, amide, urethane, urea, etc. In some embodiments, the other nitrogen-containing groups may be located in the main chain and / or side groups of the polycarbonate.

[0154] The polycarbonate described herein may have at least one (e.g., 1, 2, 3, 4, or 5) redox-sensitive groups in its repeating units (e.g., repeating units of Formula I, such as X and / or Y structures). In some embodiments, the redox-sensitive groups include both oxidation-sensitive and / or reduction-sensitive groups. In some embodiments, the redox-sensitive groups include reduction-sensitive groups.

[0155] Such redox-sensitive groups may include, for example, monosulfide bonds (-S-), monoselenoses (-Se-), disulfide bonds (-SS-), diselenoses (-Se-Se-), trisulfide bonds (-SSS-), triselenoses (-Se-Se-Se-), tetrasulfide bonds (-SSSS-), and ketethioses (-SSSS-). Where R a and R b Each is C independently 1-3 Alkyl group, wavy line One or more of the following (indicating the bonding position with adjacent atoms).

[0156] The one or more redox-sensitive groups may be located on the main chain and / or side groups of the polycarbonate. For example, at least one redox-sensitive group is located on the main chain of the polycarbonate. In some embodiments, the number of redox-sensitive groups located on the main chain of the polycarbonate in the repeating unit of Formula I may be greater than the number of redox-sensitive groups located on the side groups. In some embodiments, the repeating unit of Formula I contains no more than 5, 4, 3, 2, or 1 redox-sensitive groups on the side groups, or none at all. In some embodiments, some or all of the redox-sensitive groups in the repeating unit of Formula I are located on the main chain of the polycarbonate. In some embodiments, in the repeating unit of Formula I, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or 100% of the redox-sensitive groups are located on the main chain of the polycarbonate rather than on the side groups.

[0157] The polycarbonate may contain one or more nitrogen-containing groups and / or one or more redox-sensitive groups in its repeating units (e.g., repeating units of Formula I). ​​In some embodiments, adjacent nitrogen-containing groups and / or one or more redox-sensitive groups may be connected by a bond (such as a single bond) or a C-bond bond. 1-20 Hydroxyl groups (e.g., C) 1-16 C 1-12 C 1-10 C 1-8 C 1-6 C 1-4 Or C 1-3 It is linked to a hydrocarbon group (e.g., an alkylene group or an alkenylene group).

[0158] In some cases, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the repeating units in the polycarbonate are linked by carbonate bonds.

[0159] In some cases, the polycarbonate may be a linear polymer. In some embodiments, the polycarbonate is not a crosslinked polymer. In other embodiments, the polycarbonate is not a random copolymer.

[0160] In the polycarbonate, the degree of polymerization of the repeating unit (e.g., the repeating unit of Formula I) is not less than 2, for example, it can be 2-2000, such as 2, 3, 5, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 400, 500, 600, 800, 1000, 1200, 1500, 1800, 2000, or a range formed by any two of the above values ​​as endpoints. In some implementations, the degree of aggregation of the repeating units of Formula I can be 2-1500, 2-1000, 3-800, 3-500, 3-200, 3-150 or 3-100.

[0161] In some implementations, repeating units can be directly bonded together.

[0162] The polycarbonate may not require long polyethylene glycol (PEG) segments as part of its main chain or side groups to impart the necessary carrier properties to the polymer. Generally, PEG segments with a degree of polymerization greater than 40, 30, or 25 are considered long PEG segments. In some embodiments, the polycarbonate may not contain long PEG segments or PEG segments at all.

[0163] The polycarbonate may have a weight-average molecular weight (Mn) of 0.5 to 100 kDa. w For example, the polycarbonate may have a value of 0.5, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 22, 25, 28, 30, 35, 40, 50, 60, 70, 80, 90, 100 kDa, or a range M formed by any two of these values ​​as endpoints. w In some embodiments, the polycarbonate may have an M of 1 to 100 kDa, 2 to 100 kDa, 2 to 90 kDa, 2 to 80 kDa, 2 to 70 kDa, 2 to 60 kDa, 2 to 50 kDa, 2 to 40 kDa, 2 to 30 kDa, or 2 to 20 kDa. w .

[0164] In some embodiments, the polycarbonate has a pKa of 3.5-9.5, for example, 4.0-9.0, 4.5-8.5, 5.0-8.0, or 5.5-7.5, for example, 5.5-7.0.

[0165] In some embodiments, the polycarbonate may also be present in its salt form. For example, the salt of the polycarbonate may be formed by salting with anions such as chloride, acetate, citrate, tetrafluoroacetate, etc.

[0166] Preparation of polycarbonate

[0167] On the other hand, this article provides a method for preparing polycarbonate, comprising a polymerization reaction using a monomer having Formula II and a monomer having Formula III:

[0168] in,

[0169] X 1 and X 2 Each independently is: a key (such as a single key) or

[0170] Y 1 and Y 2 Each independently is: a key (such as a single key) or

[0171] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group;

[0172] L represents a key (such as a single key), C represents a key (such as a single key). 1-16 (For example, C) 1-12 C 1-10 C 1-8 C 1-6 C 1-4 Or C 1-3 ) Hydroxyl group (e.g., alkylene or alkenylene) or carbonate bond;

[0173] R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0174] b1 to b4 are each an independent integer from 0 to 2, for example, 0, 1 or 2;

[0175] a1 and a4 are each independently an integer from 1 to 16, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or within a range formed by any two of the above values ​​as endpoints; and

[0176] a2 and a3 are each an independent integer from 0 to 16, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or within a range formed by any two of the above values ​​as endpoints;

[0177] The restrictions are:

[0178] (1) Equation II and Equation III together have at least one X and at least one Y, and at least one X and at least one Y do not come from Equation II or Equation III at the same time;

[0179] (2) At least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0180] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0181] For the sake of brevity, for a detailed description of the (protonable) nitrogen-containing groups contained in the monomers used in the above methods, please refer to the relevant description of the polycarbonate contained in the polycarbonate nucleic acid complex above.

[0182] In some implementations, X 1 Can be with X 2 Same or different. In some implementations, Y 1 Can be with Y 2 Same or different.

[0183] In some implementations, -X 1 -Y 1 -Can be used with -X 2 -Y 2 - Same or different.

[0184] In one exemplary implementation, X 1 yes And Y 1 It is a key; X 2 It is a key and Y 2 yes L stands for bond.

[0185] The above polymerization reaction can be carried out in a certain solvent system, usually an organic solvent.

[0186] As an example, organic solvents may include, but are not limited to: dimethyl sulfoxide; C 1-10 Aliphatic hydrocarbon (including halogenated hydrocarbon) solvents, such as chloroform, dichloromethane, pentane (e.g., cyclopentane), hexane (e.g., n-hexane, cyclohexane), heptane (e.g., n-heptane, cycloheptane), octane, nonane, decane, and their isomers and mixtures; C 6-14Aromatic hydrocarbon (including halogenated aromatic hydrocarbon) solvents, such as benzene, toluene, xylene, chlorobenzene, bromobenzene, and their isomers and mixtures; ester solvents, such as methyl acetate, ethyl acetate, γ-butyrolactone and ε-caprolactone; ether solvents, such as diethyl ether, tetrahydrofuran, dioxane, and their isomers and mixtures.

[0187] The temperature and time of the reaction can be determined depending on the specific circumstances, such as the type of reactants (e.g., monomers), the required degree of polymerization, and the desired properties of the target polymer (e.g., molecular weight and molecular weight distribution).

[0188] Generally, the method has no stringent requirements on reaction temperature and is suitable for a wide range of types and large-scale production applications. For example, the reaction can be carried out in a temperature range of 20°C to 200°C. Specifically, the reaction can be carried out at 20°C, 22°C, 25°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 110°C, 120°C, 130°C, 140°C, 150°C, 160°C, 170°C, 180°C, 190°C, 200°C, or any two of these values ​​as endpoints. In some embodiments, the reaction can be carried out at temperatures not exceeding 120°C, 110°C, or 100°C, for example, not exceeding 90°C, 80°C, 70°C, 65°C, 60°C, 55°C, or 50°C. For example, when it is necessary to carry out the polymerization reaction under relatively mild reaction conditions, it may be considered to carry out the above reaction in a temperature range of 20°C to 70°C, 25°C to 70°C, 30°C to 70°C, or 35°C to 70°C.

[0189] There is no particular limitation on the reaction time. The reaction time can be determined based on the reaction temperature and the desired polymer molecular properties. For example, the reaction can last from, for instance, 4 to 96 hours. For example, the polymerization reaction can last for 4, 8, 12, 18, 24, 36, 48, 60, 72, or 96 hours, or a range of time lengths consisting of any two of these values. In some embodiments, the reaction can proceed for 4-72 hours, 4-60 hours, 4-48 hours, 8-36 hours, or 8-24 hours.

[0190] In some cases, a catalyst may be added to the reaction system to accelerate the polymerization reaction. There is no particular limit to the amount of catalyst added; it depends on the specific needs. Examples of catalysts include, but are not limited to: cesium fluoride, potassium fluoride, sodium fluoride, 4-dimethylaminopyridine (DMAP), and combinations thereof.

[0191] Optionally, the reaction products can be post-processed after the reaction, such as by precipitation, separation, drying, ultrafiltration, etc. In some exemplary embodiments, the desired reaction products can be precipitated by cooling the reaction solution, evaporating the solvent, centrifuging, etc. In some exemplary embodiments, the reaction solution can be dropped into anhydrous refrigerated diethyl ether to precipitate the reaction products. In some exemplary embodiments, the resulting precipitate can be further washed and / or dried (e.g., vacuum dried) to obtain the desired reaction products.

[0192] On the other hand, this article also provides polycarbonates prepared by the methods described herein.

[0193] Polycarbonate composites and their preparation

[0194] On the other hand, this document provides a complex formed by the polycarbonate or its salt with at least one nucleic acid, also referred to as a "polycarbonate complex".

[0195] For the purposes of this document, the nucleic acid may be a nucleic acid that needs to be delivered in a protected manner to a biological target location (e.g., cell, tissue, organ, etc.) to perform its function.

[0196] In some cases, the nucleic acids include, but are not limited to, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), peptide nucleic acid (PNA), and locked nucleic acid (LNA).

[0197] In some embodiments, the nucleic acid may be selected from the group consisting of: cDNA, mtDNA, ssDNA, dsDNA, antisense DNA, plasmid DNA, linear DNA, supercoiled DNA, siRNA, dsRNA, ssRNA, shRNA, saRNA, miRNA, rRNA, hnRNA, mRNA, tRNA, snRNA, premessenger RNA, catalytic RNA, antisense RNA, long noncoding RNA (lncRNA), and any combination thereof.

[0198] There are no particular restrictions on the length and morphology of the nucleic acid carried. Oligonucleotides of shorter length (e.g., 10 to 200 nt or bp), mRNAs of longer length, or circular plasmid DNA can all form complexes with the polycarbonate described herein and be carried to the target site. For non-limiting purposes, generally, nucleic acids with lengths not exceeding 100,000 nt or bp, 80,000 nt or bp, 60,000 nt or bp, 50,000 nt or bp, 40,000 nt or bp, 30,000 nt or bp, 25,000 nt or bp, 20,000 nt or bp, 18,000 nt or bp, 15,000 nt or bp, or 12,000 nt or bp can be selected as the carrier. In some implementations, the length can be selected from 1 to 100,000 nt or bp, for example, 1, 2, 5, 10, 20, 30, 40, 50, 80, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 6 Nucleic acids of 000, 7000, 8000, 9000, 10000, 12000, 15000, 18000, 20000, 25000, 30000, 40000, 50000, 60000, 70000, 80000, 90000, 100000 nt or bp, or within a range formed by any two of the above values ​​as endpoints, may be used as carriers.

[0199] In some embodiments, the nucleic acid may be of natural or synthetic origin. In some embodiments, the nucleic acid may include single-stranded and / or double-stranded nucleic acids. It should be understood that the nucleotide sequence of certain single-stranded nucleic acids (e.g., mRNA) may contain self-complementary nucleotide segments that can fold back through base pairing to form a partially double-stranded structure. Such nucleic acids with partially double-stranded segments also fall within the scope of the term "single-stranded nucleic acid" as used in this invention.

[0200] In some implementations, the nucleic acid can be endogenous or exogenous.

[0201] In the complex, the composite mass ratio of polycarbonate to nucleic acid can range from 0.1:1 to 500:1. For example, the composite mass ratio can be 0.1:1, 0.5:1, 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 8:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, etc. 50:1, 60:1, 70:1, 80:1, 90:1, 110:1, 120:1, 150:1, 180:1, 200:1, 220:1, 250:1, 280:1, 300:1, 320:1, 350:1, 380:1, 400:1, 420:1, 450:1, 480:1, 500:1, or within a range formed by any two of the above values ​​as endpoints.

[0202] The composite may have an average particle size between 10 and 1000 nm, for example 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 31 0, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000 nm, or the average particle size within a range formed by taking any two of the above values ​​as endpoints.

[0203] On the other hand, this article provides a method for preparing a polycarbonate-nucleic acid complex, the method comprising:

[0204] - Provide the polycarbonate or a salt thereof, and at least one nucleic acid; and

[0205] - Mix the polycarbonate or its salt with the at least one nucleic acid.

[0206] Generally, the mixing is carried out in an aqueous medium, preferably an acidic aqueous medium. In some embodiments, the aqueous medium may have a pH not higher than 7.4, for example, not higher than 6.5, 6, 5.5, or 5. In some embodiments, the aqueous medium may have a pH from 1 to 7.4, for example, 2 to 7, 2 to 6.5, 2 to 6, 2 to 5.5, or 2 to 5. In other embodiments, the mixing may be carried out at a pH below the pKa of the polycarbonate or its salt. For example, the complex can be prepared by mixing the above-described aqueous medium containing the polycarbonate or its salt with nucleic acids.

[0207] In some embodiments, the complex is in an aqueous medium with a pH not higher than 7.4, for example, not higher than 6.5, 6, 5.5, or 5. In some embodiments, the complex is in an aqueous medium with a pH of 2 to 7, 2 to 6, 2 to 5.5, or 2 to 5.

[0208] In some exemplary embodiments, the aqueous medium may be an acidic aqueous buffer solution, such as phosphate buffer, acetate-sodium acetate buffer, citrate buffer, acetate-ammonium acetate buffer, oxalate buffer, etc. In some embodiments, the acidic aqueous medium may contain an organic solvent, such as no more than 10% w / w DMSO, if necessary.

[0209] Mixing can be achieved by adding an aqueous solution of nucleic acid dropwise to an aqueous medium containing dissolved polycarbonate, or vice versa. Thorough mixing can also be promoted using methods such as shaking, vortexing, stirring, inverting, and microfluidics.

[0210] The mixing can be carried out at a temperature that has no significant impact on the structure and properties of both the polycarbonate and the nucleic acid (e.g., the degree of damage is less than 10% of the original level). In some embodiments, the mixing can be carried out at room temperature.

[0211] After mixing, the resulting mixture can be incubated for a period of time to allow for better formation of the polycarbonate nucleic acid complex.

[0212] Incubation can be performed at a temperature that has no significant effect on the structure and properties of both the polycarbonate and the nucleic acid (e.g., the degree of damage is less than 10% of the original level). In some embodiments, incubation can be performed at room temperature. In some embodiments, the resulting mixture is incubated at room temperature for 5 to 60 minutes, for example 10 to 40 minutes, or for example 10 to 30 minutes, to obtain the polycarbonate complex.

[0213] On the other hand, this article provides a polycarbonate composite prepared by the above preparation method.

[0214] Compositions and Products

[0215] On the other hand, this document provides a composition comprising the polycarbonate or a salt thereof as described herein, or a polycarbonate complex as described herein.

[0216] In some embodiments, the composition may be a pharmaceutical composition.

[0217] In some embodiments, the composition comprises a pharmaceutically acceptable carrier and at least one polycarbonate described herein.

[0218] In other embodiments, the composition comprises a pharmaceutically acceptable carrier and at least one polycarbonate complex described herein.

[0219] The delivery vehicle includes liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials involved in carrying or transporting the polycarbonate or polycarbonate complex to an organ or part of the body. Each delivery vehicle must be "acceptable" in terms of compatibility with other components of the formulation and safety for the patient.

[0220] Some examples of substances that can be used as pharmaceutically acceptable carriers include: polysaccharides and their derivatives, proteins, lipids, inorganic fillers, solvents, auxiliaries (such as injection diluents, buffers or carrier solutions), excipients, esters, polymers, etc. Specifically, this may include: cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; polysaccharides, such as powdered astragalus gum, alginic acid, agar, and malt; proteins, such as gelatin and collagen; inorganic fillers / buffers, such as talc, calcium carbonate, calcium phosphate, silica, magnesium hydroxide, and aluminum hydroxide; aqueous solvents / diluents, such as water (e.g., pyrogen-free water), isotonic saline, Ringer's solution, pH buffer solution, and ethanol; hydrophilic polyols, such as glycols (e.g., propylene glycol), glycerol, sorbitol, and mannitol; oil phase carriers / hydrophobic solvents, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; ester solvents / oil-based adjuvants, such as ethyl oleate and ethyl laurate; excipients, such as cocoa butter and suppository waxes; other polymeric materials, such as polyesters, polyamides, polycarbonates, and / or polyanhydrides; and other non-toxic and compatible substances used in pharmaceutical formulations. Pharmaceutically acceptable delivery vehicles for use in conjunction with active pharmaceutical ingredients are well known in the art. They can be used in the compositions described herein, provided that conventionally used pharmaceutically acceptable delivery vehicles do not adversely react with the active ingredient (such as the polycarbonate complexes described herein). The compositions described herein may also contain other active substances as desired.

[0221] It should be understood that the relative amounts of the active ingredient, pharmaceutically acceptable carrier, and / or other components in the composition are variable depending on the application suitable for the active ingredient (such as the polycarbonate complex described herein) and may depend on multiple factors, such as the age, sex, weight, physical condition, and route of administration of the composition to the target recipient. As an example, the composition may contain 0.1% to 100% (w / w) of the active ingredient (such as the polycarbonate complex described herein) by weight, for example 0.1%, 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% (w / w), or a range of the active ingredient (such as the polycarbonate complex described herein) formed by any two of the above values ​​as endpoints.

[0222] In some embodiments, the composition may have a pH not higher than 7.4, for example, not higher than 6.5, not higher than 6, not higher than 5.5, or not higher than 5. In some embodiments, the composition may have a pH from 1 to 7.4, for example, 2 to 7, 2 to 6.5, 2 to 6, 2 to 5.5, or 2 to 5.

[0223] In some embodiments, the composition may also contain other active substances.

[0224] On the other hand, this article provides a product comprising:

[0225] The polycarbonate composites described in this article;

[0226] The compositions described herein; or

[0227] The polycarbonate or its salt and at least one nucleic acid described herein.

[0228] In some embodiments, the product is used to deliver at least one nucleic acid to a cell or object. In some embodiments, the product is used to express at least one nucleic acid in a cell or object.

[0229] In some embodiments, the product comprises polycarbonate or a salt thereof, and at least one nucleic acid. In such embodiments, the polycarbonate or a salt thereof and the nucleic acid are physically separated from each other in the product (e.g., packaged separately) for mixing and forming a complex immediately before use.

[0230] In some embodiments, the product comprises the polycarbonate or its salt, at least one nucleic acid, a polycarbonate nucleic acid complex, or a composition comprising one or more of these, packaged in single or multiple unit doses.

[0231] In some embodiments, the product may comprise the polycarbonate or a salt thereof, at least one nucleic acid, a polycarbonate nucleic acid complex, or a composition comprising one or more of these, packaged in a convenient fraction of a single unit dose (e.g., half or one-third of the dose).

[0232] In some embodiments, the product may also include one or more other components or ingredients, such as instructions for use; adjuvants, such as markers, other therapeutic agents, etc.; and operating devices, such as suction devices, mixing devices, measuring devices, sealing devices, etc.

[0233] Usage and Purpose

[0234] On the other hand, a method for delivering at least one nucleic acid to a cell or object is provided, the method comprising:

[0235] - Provide a complex or composition comprising the complex described herein, the complex being formed by combining the at least one nucleic acid with a polycarbonate or a salt thereof described herein; and

[0236] - To bring the cell or the object into contact with the complex.

[0237] On the other hand, a method for expressing at least one nucleic acid in a cell or object is provided, the method comprising:

[0238] - Provide a complex or composition comprising the complex described herein, the complex being formed by combining the at least one nucleic acid with a polycarbonate or a salt thereof described herein; and

[0239] - To bring the cell or the object into contact with the complex.

[0240] On the other hand, it provides the use of the polycarbonate or its salt, the polycarbonate complex or the composition for delivering at least one nucleic acid to a cell or object.

[0241] On the other hand, the polycarbonate or its salt, the polycarbonate complex or the composition are provided for use in enabling cells or objects to express at least one nucleic acid.

[0242] On the other hand, the polycarbonate or its salt is provided for use in the preparation of a product for delivering at least one nucleic acid to a cell or object.

[0243] On the other hand, the polycarbonate or its salt is provided for use in the preparation of a product for causing cells or objects to express at least one nucleic acid.

[0244] Generally, delivering or expressing at least one nucleic acid to or in a subject can be delivering or expressing the at least one nucleic acid to or in one or more body parts of the subject. The one or more body parts may include, for example, organs, muscles, subcutaneous tissue, joint cavities, bones and their microenvironments, blood, lymph, and other sites where the at least one nucleic acid needs to be delivered.

[0245] In some embodiments, the cells may be ex vivo cells. In some embodiments, the cells may be adherent or suspension cultured cells. In some embodiments, the cells may be cells derived from or derived from organs or tissues. In some embodiments, the cells may be primary cells or immortalized cell lines.

[0246] In some embodiments, the object may be an invertebrate or a vertebrate. In some embodiments, the object may include mammals, such as non-human mammals and humans.

[0247] In some embodiments, the contact may be sustained for a duration sufficient to allow the complex or at least one nucleic acid contained therein to enter one or more body parts of a cell or object, for example, 10 minutes or longer.

[0248] In some embodiments, the at least one nucleic acid may be an endogenous or exogenous nucleic acid, for example, an endogenous or exogenous nucleic acid that is required by the subject or beneficial to their health.

[0249] In some embodiments, the delivery or expression is in vitro or in vivo.

[0250] In some embodiments, the delivery may be carried out via intra-arterial, intravenous, intraperitoneal, extra-gastric, intramuscular, subcutaneous, oral, inhalation, or local routes.

[0251] The development of polycarbonate described in this article opens up a new pathway for the biological delivery of nucleic acids. Effective delivery of nucleic acids to biological target sites has always been a significant challenge, as nucleic acids are not only easily affected by cationic interference during delivery, leading to inactivation or degradation, but also require overcoming the cell membrane barrier to enter the cell nucleus.

[0252] The polycarbonate nanocarriers described herein can carry a wide range of nucleic acids with varying molecular weights and lengths, exhibiting extremely high adaptability for biodelivery. As a carrier, the polycarbonate provides effective protection, promotes transmembrane transport, and degrades promptly upon arrival at a specific environment (such as the target biological location), thereby rapidly releasing the nucleic acids for optimal utilization. The degradation products of the polycarbonate (e.g., CO2, thiols, etc.) are biosafe, significantly eliminating the toxic side effects of existing carriers. Furthermore, the preparation process of the polycarbonate is simple and user-friendly, with controllable product quality, making it suitable for large-scale production and subsequent biological experiments and industrial applications.

[0253] The unique properties of the polycarbonate nanomaterials make them suitable as a next-generation nucleic acid delivery carrier for safe and efficient nucleic acid delivery.

[0254] Implementation methods / combinations

[0255] Embodiment A1: A polycarbonate or a salt thereof, said polycarbonate having repeating units of Formula I:

[0256] in,

[0257] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group, with the constraint that at least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0258] L is a key, C 1-16 Hydroxyl or carbonate bond;

[0259] R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0260] a1 and a4 are each independent integers from 1 to 16;

[0261] a2 and a3 are each independent integers from 0 to 16; and

[0262] b1 to b4 are each an independent integer between 0 and 2.

[0263] Embodiment A2: The polycarbonate or its salt according to Embodiment A1, wherein the at least one nitrogen-containing group is located in the main chain.

[0264] Embodiment A3: The polycarbonate or its salt according to Embodiment A1, wherein at least one nitrogen atom of the at least one nitrogen-containing group is located in the main chain.

[0265] Embodiment A4: The polycarbonate or its salt according to Embodiment A1, wherein the at least one protonable nitrogen-containing group is located in the main chain.

[0266] Embodiment A5: The polycarbonate or its salt according to Embodiment A1, wherein at least one nitrogen atom of the at least one protonable nitrogen-containing group is located in the main chain.

[0267] Embodiment A6: The polycarbonate or its salt according to Embodiment A1, wherein the at least one protonable nitrogen-containing group has a group selected from the group consisting of amino groups, nitrogen-containing cyclic groups, and combinations thereof.

[0268] Embodiment A7: The polycarbonate or its salt according to claim A6, wherein the amino group is selected from the group consisting of secondary amino groups, tertiary amino groups, quaternary amino groups, and combinations thereof.

[0269] Embodiment A8: The polycarbonate or its salt according to claim A6, wherein the nitrogen-containing cyclic group is selected from the group consisting of 3 to 10 saturated or unsaturated aliphatic or aromatic heterocyclic groups having at least one cyclic nitrogen atom, 5 to 20 fused heterocyclic groups having at least one cyclic nitrogen atom and 3 to 10 saturated or unsaturated aliphatic or aromatic heterocyclic groups fused together or with aliphatic or aromatic carbon rings, and combinations thereof.

[0270] Embodiment A9: The polycarbonate or its salt according to claim A1, wherein at least one of the at least one redox-sensitive groups is located in the main chain.

[0271] Implementation A10: The polycarbonate or its salt according to Implementation A1, wherein the at least one redox sensitive group comprises one or more of the following: monosulfide bond, monoselenide bond, disulfide bond, diselenide bond, trisulfide bond, triselenide bond, tetrasulfide bond, and ketethiol bond.

[0272] Embodiment A11: The polycarbonate or its salt according to Embodiment A1, wherein the polycarbonate has a weight-average molecular weight (M) of 0.5 to 100 kDa. w ).

[0273] Embodiment A12: The polycarbonate or salt thereof according to Embodiment A1, wherein the polycarbonate has a pKa of 3.5-9.5.

[0274] Implementation Method B1: A method for preparing polycarbonate, comprising a polymerization reaction using a monomer having Formula II and a monomer having Formula III:

[0275] In Equations II and III,

[0276] X 1 and X 2 Each independently is: key or

[0277] Y 1 and Y 2 Each independently is: key or

[0278] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group;

[0279] L is a key, C 1-16 Hydroxyl or carbonate bond;

[0280] R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups, and substituted or unsubstituted 5- to 14-membered heteroaryl groups;

[0281] b1 to b4 are each an independent integer from 0 to 2;

[0282] a1 and a4 are each independent integers from 1 to 16; and

[0283] a2 and a3 are each independent integers from 0 to 16;

[0284] The restrictions are:

[0285] (1) Equation II and Equation III together have at least one X and at least one Y, and at least one X and at least one Y do not come from Equation II or Equation III at the same time;

[0286] (2) At least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0287] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0288] Implementation method B2: According to the method of implementation method B1, wherein X 1 yes And Y 1 It is a key; X 2 It is a key and Y 2 yes L stands for bond.

[0289] Implementation C1: A complex of polycarbonate or a salt thereof according to Implementation A1 with at least one nucleic acid.

[0290] Implementation C2: The polycarbonate nucleic acid complex according to Implementation C1, wherein the at least one nucleic acid is selected from the group consisting of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), peptide nucleic acid (PNA), locked nucleic acid (LNA), and any combination thereof.

[0291] Implementation C3: The polycarbonate nucleic acid complex according to Implementation C1, wherein the at least one nucleic acid includes single-stranded nucleic acid and / or double-stranded nucleic acid.

[0292] Implementation C4: The polycarbonate nucleic acid complex according to Implementation C1, wherein the composite mass ratio of the polycarbonate or its salt to the at least one nucleic acid ranges from 0.1:1 to 500:1.

[0293] Embodiment C5: The polycarbonate nucleic acid complex according to Embodiment C1, wherein the complex has an average particle size between 10 and 1000 nm.

[0294] Embodiment D1: A composition comprising the polycarbonate or a salt thereof as described in Embodiment A1, or the complex as described in Embodiment C1.

[0295] Implementation D2: The composition according to Implementation D1, wherein the composition is a pharmaceutical composition.

[0296] Implementation D3: The composition according to Implementation D2 further comprises at least one pharmaceutically acceptable carrier.

[0297] Implementation Method D4: The composition according to Implementation Method D3, wherein the pharmaceutically acceptable carrier is selected from the group consisting of: polysaccharides and their derivatives, proteins, lipids, inorganic fillers, solvents, auxiliaries (such as injection diluents, buffers or carrier solutions), excipients, esters, polymers, and any combination thereof.

[0298] Embodiment D5: The composition according to Embodiment D1, wherein the composition comprises from 0.1% to 100% (w / w) of the polycarbonate or a salt thereof, or the complex thereof, by weight.

[0299] Embodiment D6: The composition according to Embodiment D1, wherein the composition has a pH not higher than 7.4, not higher than 6.5, not higher than 6, not higher than 5.5 or not higher than 5.

[0300] Embodiment D7: The composition according to Embodiment D1, wherein the composition further comprises other active substances.

[0301] Implementation D8: The composition according to Implementation D1, wherein the composition is formulated as an injection, oral preparation, topical preparation, inhalant or implant, such as a solution, emulsion, suspension, gel, ointment, patch, capsule, tablet, powder inhaler, aerosol and other dosage forms.

[0302] Implementation method E1: A product comprising:

[0303] -The polycarbonate or its salt thereof and at least one nucleic acid according to embodiment A1;

[0304] -The composition according to embodiment D1; or

[0305] -The complex according to embodiment C1.

[0306] Implementation E2: The product according to Implementation E1 is used to deliver at least one nucleic acid to a cell or object, or to express at least one nucleic acid in a cell or object.

[0307] Implementation E3: The product according to Implementation E1 comprises the polycarbonate or its salt physically separated from each other and at least one nucleic acid.

[0308] Implementation E4: The product according to implementation E1, wherein the product further comprises one or more other components or parts selected from the group consisting of: instructions for use, auxiliary agents, operating instruments, and any combination thereof.

[0309] Implementation F1: A method for delivering at least one nucleic acid to a cell or object, the method comprising:

[0310] - Provides a polycarbonate nucleic acid complex according to embodiment C1, the complex being formed by combining the at least one nucleic acid with the polycarbonate or a salt thereof; and

[0311] - To bring the cell or the object into contact with the complex.

[0312] Implementation method G1, a method for expressing at least one nucleic acid in a cell or object, the method comprising:

[0313] - Provides a polycarbonate nucleic acid complex according to embodiment C1, the complex being formed by combining the at least one nucleic acid with the polycarbonate or a salt thereof; and

[0314] - To bring the cell or the object into contact with the complex.

[0315] Implementation H1: The use of polycarbonate or its salt as described in Implementation A1 as a nucleic acid carrier for delivering the nucleic acid to a cell or object.

[0316] Embodiment I1, the use of the polycarbonate or its salt according to Embodiment A1, the polycarbonate nucleic acid complex according to Embodiment C1, or the composition according to Embodiment D1 for delivering at least one nucleic acid to a cell or object.

[0317] Embodiment J1, the use of the polycarbonate or its salt according to Embodiment A1, the polycarbonate nucleic acid complex according to Embodiment C1, or the composition according to Embodiment D1 for expressing at least one nucleic acid in a cell or object.

[0318] Implementation K1: Use of the polycarbonate or its salt according to Implementation A1, the polycarbonate nucleic acid complex according to Implementation C1, or the composition according to Implementation D1 in the preparation of a product for delivering at least one nucleic acid to a cell or object.

[0319] Embodiment L1: Use of the polycarbonate or its salt according to Embodiment A1, the polycarbonate nucleic acid complex according to Embodiment C1, or the composition according to Embodiment D1 in the preparation of a product for expressing at least one nucleic acid in a cell or object.

[0320] Implementation method M1: The method according to implementation method F1 or G1 or the use according to any one of implementation methods H1 to L1, wherein the cell is an isolated cell.

[0321] Implementation method M2: The method according to implementation method F1 or G1 or the use according to any one of implementation methods H1 to L1, wherein the object is an invertebrate or a vertebrate.

[0322] Implementation method M3: The method according to implementation method F1 or G1 or the use according to any one of implementation methods H1 to L1, wherein the object is a mammal.

[0323] Implementation M4: The method according to implementation F1 or G1, wherein the contact is sustained for a duration sufficient to allow the complex or at least one nucleic acid contained therein to enter one or more body parts of a cell or object.

[0324] Implementation method M5: The method according to implementation method F1, or the use according to implementation method H1, I1 or K1, wherein the delivery is in vivo delivery or in vitro delivery.

[0325] Implementation method M6: The method according to implementation method F1, or the use according to implementation method H1, I1 or K1, wherein the delivery is carried out via intra-arterial, intravenous, intraperitoneal, extra-gastric, intramuscular, subcutaneous, oral, inhalation or local routes.

[0326] Example

[0327] The following embodiments further describe and illustrate implementations according to the present invention. These embodiments are given for illustrative purposes only and should not be construed as limiting the invention, as many variations may be made without departing from the spirit and scope of the invention.

[0328] Materials and Instruments

[0329] The instrument information used in the embodiments is summarized in Table 1 below:

[0330] Table 1. Instruments used in the examples

[0331] The material information used in the embodiments is summarized in Table 2 below:

[0332] Table 2. Materials used in the examples

[0333] Example 1: Monomer Preparation and Characterization

[0334] Preparation Example A1: 2,2'-((3-methoxypropyl)azadiyl)bis(1-ethanol) (A013)

[0335] Diethanolamine (1.46 g, 9.54 mmol), 1-bromo-3-methoxypropane (1.56 g, 14.84 mmol), and anhydrous potassium carbonate (1.58 g, 11.44 mmol) were weighed out. Dry acetonitrile (30 mL) was then added, and the reaction was carried out at 85 °C for 18 hours. After the reaction mixture returned to room temperature, the acetonitrile was removed by vacuum distillation. Saturated brine (85 mL) was added to the evaporated residue, and the mixture was extracted with ethyl acetate (170 mL, 3X). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation, and the residue was dried to obtain monomer A013 (1.20 g), yield 71.1%.

[0336] 1 H NMR (700MHz, CDCl3): δ3.62(t, CH2N(CH2CH2OH)2), 3.50(t, CH3OCH2CH2), 3.33(s, CH3OCH2CH2), 2.68(m, CH3OCH2CH2CH2N(CH2CH2OH)2), 1.78(m, CH3OCH2CH2CH2N).

[0337] HRMS(ESI,m / z):C8H 19 NO3, [M+H] + Calculated value: 178.1443; Measured value: 178.1428.

[0338] Preparation Example A2: 2,2'-((3-(pyrrolidone-1-yl)propyl)azadiyl)bis(1-ethanol)(A014)

[0339] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 1-(3-chloropropyl)pyrrolidine, the extraction solvent was replaced with dichloromethane, and the product was purified by column chromatography to obtain monomer A014 with a yield of 65.0%.

[0340] 1 H NMR (700MHz, CDCl3): δ3.62(t,NCH2CH2OH)2), 2.68-2.63(m,CH2CH2CH2CH2N(CH2CH2OH)2), 2.58(m,NCH2CH2CH2N), 1.83(m,CH2NCH2CH2CH2CH2), 1.67(m,NCH2CH2CH2N).

[0341] HRMS(ESI,m / z):C 11 H 24 N₂O₂, [M+H] + Calculated value: 217.1916; Measured value: 217.1935.

[0342] Preparation Example A3: 2,2'-((3-(piperidin-1-yl)propyl)azadiyl)bis(1-ethanol) (A015)

[0343] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 1-(3-bromopropyl)piperidine hydrobromide, and the extraction solvent was replaced with dichloromethane instead of ethyl acetate. The monomer A015 obtained after post-treatment had a yield of 76.0%.

[0344] 1 H NMR(700MHz,CDCl3):3.61(t,CH2NCH2CH2OH)2),2.62(t,CH2N(CH2CH2OH)2),2.61(t,CH2N(CH2CH2OH)2),2.47(m,CH2CH2N CH2CH2CH2CH2CH2),1.65(m,CH2CH2NCH2CH2CH2CH2CH2),1.63(m,CH2CH2NCH2CH2CH2CH2CH2),1.44(s,NCH2CH2CH2CH2CH2).

[0345] HRMS(ESI,m / z):C 12 H 26 N₂O₂, [M+H] + Calculated value: 231.2073; Measured value: 231.2083.

[0346] Preparation Example A4: 2,2'-((3-morpholinopropyl)azadiyl)bis(1-ethanol) (A016)

[0347] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 4-(3-bromopropyl)morpholine, the extraction solvent was replaced with dichloromethane, and the product was purified by column chromatography to obtain monomer A016 with a yield of 62.1%.

[0348] 1 H NMR (700MHz, CDCl3): δ3.73(t,CH2NCH2CH2OCH2CH2), 3.62(t,CH2NCH2CH2OH)2), 2.61(t,CH2N(CH2CH2OH)2), 2.47(NCH2CH2CH2N), 1.67(m,NCH2CH2CH2N).

[0349] HRMS(ESI,m / z):C 11 H 24 N₂O₃, [M+H] +Calculated value: 233.1865; Measured value: 233.1883.

[0350] Preparation Example A5: 2,2'-((3-(1H-pyrrolo-1-yl)propyl)azadiyl)bis(1-ethanol) (A017)

[0351] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 1-(3-bromopropyl)pyrrole, and the extraction solvent was replaced with dichloromethane instead of ethyl acetate. The yield of monomer A017 after post-treatment was 76.0%.

[0352] 1 H NMR (700MHz, CDCl3): δ6.67(t,NCHCH),6.14(t,NCHCH),3.94(t,CH2NCH2CH2OH)2),3.60(t,C H2NCH2CH2OH)2),2.65(t,CH2N(CH2CH2OH)2),2.54(t,NCH2CH2CH2N),1.99(m,NCH2CH2CH2N).

[0353] HRMS(ESI,m / z):C 11 H 20 N₂O₂, [M+H] + Calculated value: 213.1603; Measured value: 213.1612.

[0354] Preparation Example A6: 2,2'-((3-(4-methylpiperazin-1-yl)propyl)azadiyl)bis(1-ethanol) (A018)

[0355] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 1-(3-chloropropyl)-4-methylpiperazine, the extraction solvent was replaced with dichloromethane, and the product was purified by column chromatography to obtain monomer A018 in 46.5% yield.

[0356] 1 H NMR (700MHz, CDCl3): δ3.62(t,J=5.3Hz,4H),2.63–2.58(m,6H),2.46(t,J=6.4Hz,2H),2.29(s,3H),1.66(q,J=6.4Hz,2H).

[0357] HRMS(ESI,m / z):C 12 H 27 N3O2, [M+H] + Calculated value: 246.2182; Measured value: 246.2196.

[0358] Preparation Example A7: 2,2'-((3-(dimethylamino)propyl)azadiyl)bis(1-ethanol) (A019)

[0359] The process was the same as described in Preparation Example A1, except that 1-bromo-3-methoxypropane was replaced with 3-bromo-N,N-dimethyl-1-propanamine hydrobromide, the extraction solvent was replaced with dichloromethane, and the product was purified by column chromatography to obtain monomer A019 with a yield of 36.6%.

[0360] 1 H NMR (700MHz, CDCl3): δ3.55(t,CH2NCH2CH2OH)2),2.58(t,CH2N(CH2CH2OH)2),2.52(t,C H2N(CH2CH2OH)2),2.37(m,CH2N(CH3)2),2.18(m,CH2N(CH3)2),1.57(m,NCH2CH2CH2N).

[0361] HRMS(ESI,m / z):C9H 22 N₂O₂, [M+H] + Calculated value: 191.1760; Measured value: 191.1749.

[0362] Preparation Example A8: 2,2'-Thiobis(1-ethanol) (A025)

[0363] Weigh 10.0 g (128 mmol, 1 eq) of 2-mercaptoethanol, 12.4 g (141 mmol, 1.1 eq) of ethylene carbonate, and 417.0 mg (1.28 mmol, 0.01 eq) of anhydrous cesium carbonate into a reaction flask and heat directly to 120 °C for 20 minutes. After the reaction solution returns to room temperature, it is directly passed through a reverse-phase C2 reactor. 18 Column (SW-5202-080-SP(H), (C 18 The mobile phase consisted of water and acetonitrile containing 0.1% TFA (20-30 μM, Sante Technology (Changzhou) Co., Ltd.). The product after adding 10% acetonitrile was collected, rotary evaporated, and freeze-dried to give a clear oily substance, 2,2'-thiobis(1-ethanol) (9.3 g), with a yield of 59.5%. The product was confirmed by thin-layer chromatography: mobile phase dichloromethane:methanol = 1:1, Rf = 0.1.

[0364] 1 H NMR (600MHz, CDCl3): δ3.75 (d, J = 11.1Hz, 2H), 3.70 (t, J = 6.1Hz, 4H), 2.69 (t, J = 6.1Hz, 4H).

[0365] Preparation Example A9: 2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethane-1-ol))(A027)

[0366] N,N'-thiodiphthalimide (25.3 g, 1 eq) and 2-mercaptoethanol (5.3 g, 0.9 eq) were dissolved in toluene (100 mL) and heated at 80 °C for 1 hour. The reaction mixture was cooled to room temperature and filtered. The white solid was washed three times with DCM, and the filtrate and washings were mixed and evaporated to dryness to obtain a crude solid. The crude solid was subjected to column chromatography (PE:EA = 1:0-1:1, Rf = 0.4) to give the product 2-((2-hydroxyethyl)dithioalkyl)isoindoline-1,3-dione (15 g, white solid, 75.3%). 2-((2-hydroxyethyl)dithioalkyl)isoindoline-1,3-dione (15 g, 1 eq) and 2-mercaptoethanol (4.14 g, 0.9 eq) were dissolved in toluene (50 mL) and stirred overnight at room temperature. The reaction mixture was filtered, washed three times with dichloromethane, and filtered through a reverse-phase C10 ... 18 The chromatographic column was used to collect the fraction containing 5%-10% acetonitrile. The fraction was then rotary evaporated and freeze-dried to obtain a transparent oily substance, 2,2'-trithionedidylbis(1-ethanol)A027 (6 g, 54.8%). The product was confirmed by thin-layer chromatography with a mobile phase of petroleum ether:ethyl acetate = 1:1 and Rf = 0.4.

[0367] 1 H NMR (700MHz, CDCl3): δ2.95(t,SCH2CH2OH),4.05(q,SCH2CH2OH),3.15(t,SCH2CH2OH)

[0368] HRMS(ESI,m / z):C7H 12 O4S2,[M+Na] + Calculated value: 247.0075; Measured value: 247.0103.

[0369] Preparation Example A10: 2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethane-1-ol))(A028)

[0370] (1) Dry HCl was bubbled into a mixture of mercaptoacetic acid (16.35 g, 177.5 mmol) and acetone (21.4 g, 369.1 mmol), and the reaction was carried out at room temperature for 6 hours. The reaction mixture was then cooled in an ice bath to allow crystals to crystallize. The precipitated solid was collected and washed with n-hexane and cold water, and dried under vacuum to give intermediate 2,2'-(propane-2,2-diylbis(thionyl))bis(propionic-1-acid) (15.23 g), with a yield of 36.8%.

[0371] 1H NMR (700MHz, DMSO-d6): δ12.63(s,CH2COOH),3.37(s,SCH2COOH),1.54(s,(CH3)2CSS).

[0372] HRMS(ESI,m / z):C7H 12 O4S2,[M+Na] + Calculated value: 247.0075; Measured value: 247.0103.

[0373] (2) Under nitrogen atmosphere, the intermediate obtained in (1) (10.00 g, 44.59 mmol) was dissolved in dry tetrahydrofuran (50 mL) and added dropwise to a tetrahydrofuran solution of LiAlH4 (100 mL) through a constant pressure dropping funnel at 0 °C for 1 hour. After the addition was complete, the reaction solution was transferred to room temperature and reacted for 30 minutes, then the reaction was continued at 52 °C for 2 hours. After the reaction was completed, the reaction solution was cooled to 0 °C, and water (6 mL), 15% NaOH (6 mL), and water (15 mL) were added successively, and the mixture was stirred for 30 minutes. Then, the mixture was extracted with diethyl ether (3×, 500 mL), and the organic layer was collected and dried with anhydrous sodium sulfate. The organic layer was distilled under reduced pressure to remove the solvent and dried to obtain monomer A028 (5.92 g), with a yield of 67.6%.

[0374] 1 H NMR (700MHz, DMSO-d6): δ4.82(t,SCH2CH2OH),3.52(q,SCH2CH2OH),2.65(t,SCH2CH2OH),1.53(s,(CH3)2CSS).

[0375] HRMS(ESI,m / z):C7H 16 O2S2,[M+Na] + Calculated value: 219.0489; Measured value: 219.0512.

[0376] Preparation Example A11: 3,3'-(propane-2,2-diylbis(thioalkyldiyl))bis(prop-1-ol) (A029)

[0377] (1) The process of preparing Example A8 is referenced, except that mercaptoacetic acid in (1) is replaced with 3-mercaptopropionic acid. Additionally, the intermediate added in step (2) is the one synthesized in Example (1) of this invention. The yields of the intermediate and monomer A029 obtained after post-treatment were 61.5% and 94.0%, respectively.

[0378] Intermediate:

[0379] 1H NMR (700MHz, DMSO-d6): δ12.26(s,CH2COOH),2.72(t,SCH2CH2COOH),2.48(t,SCH2CH2COOH),1.51(s,(CH3)2CSS).

[0380] HRMS(ESI,m / z):C9H 16 O4S 2, [M+Na] + Calculated value: 275.0388; Measured value: 275.0381.

[0381] Monomer A029:

[0382] 1 H NMR (700MHz, DMSO-d6): δ4.49(t,SCH2CH2CH2OH),3.42(q,SCH2CH2CH2OH),2.59(t,SCH2CH2CH2OH),1.62(m,SCH2CH2CH2OH),1.51(s,(CH3)2CSS).

[0383] HRMS(ESI,m / z):C9H 20 O2S2,[M+Na] + Calculated value: 247.0802; Measured value: 247.0799.

[0384] Preparation Example A12: 3-((2-hydroxyethyl)(methyl)amino)prop-1-ol (A084)

[0385] Weigh 2.50 g (28.05 mmol) of 3-(aminomethyl)-1-propanol, 7.0100 g (56.09 mmol) of 2-bromoethanol, and 9.30 g (67.31 mmol) of anhydrous potassium carbonate, then add 75 mL of dry acetonitrile and react at 85 °C for 10 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A0084 (1.62 g) was obtained in a yield of 43.3%.

[0386] 1H NMR (400MHz, CDCl3): δ3.76(t,HOCH2CH2NCH3),3.66(t,HOCH2CH2CH2NCH3),3.42(s,HOCH2CH2CH2NCH3),2. 61(t,HOCH2CH2NCH3),2.55(t,HOCH2CH2CH2NCH3),2.28(t,HOCH2CH2CH2NCH3),1.72(m,HOCH2CH2CH2NCH3).

[0387] HRMS(ESI,m / z):C6H 16 NO 2； [M+H] + Calculated value: 134.1181; Measured value: 134.1175.

[0388] Preparation Example A13: 5-((3-hydroxypropyl)(methyl)amino)pentan-1-ol (A086)

[0389] Weigh 3-(aminomethyl)-1-propanol (3.50 g, 39.26 mmol), 5-bromo-1-pentanol (11.15 g, 66.75 mmol), and anhydrous potassium carbonate (11.07 g, 80.10 mmol), then add 100 mL of dry acetonitrile and react at 85 °C for 3 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A086 (3.31 g) was obtained, with a yield of 48.1%.

[0390] 1 H NMR(400MHz,CDCl3): δ3.79(t,HOCH2CH2CH2CH2CH2NCH3),3.61(t,HOCH2CH2CH2NCH3),2.58(t,HOCH2CH2CH2NCH3),2.37(t,HOCH2CH2CH2CH2CH2NCH3),2.2 2(s,HOCH2CH2CH2CH2CH2NCH3),1.69(m,HOCH2CH2CH2CH2CH2NCH3),1.54(m,HOCH2CH2CH2CH2CH2N(CH3)CH2CH2CH2OH),1.37(m,HOCH2CH2CH2CH2CH2NCH3).

[0391] HRMS(ESI,m / z):C9H 22 NO 2； [M+H] + Calculated value: 176.1651; Measured value: 176.1654.

[0392] Preparation Example A14: 6-((3-hydroxypropyl)(methyl)amino)hexane-1-ol (A087)

[0393] Weigh 3-(aminomethyl)-1-propanol (3.50 g, 39.26 mmol), 6-bromo-1-hexanol (12.09 g, 66.75 mmol), and anhydrous potassium carbonate (11.07 g, 80.10 mmol), then add 100 mL of dry acetonitrile and react at 85 °C for 2 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A087 (3.95 g) was obtained in a yield of 53.2%.

[0394] 1 H NMR (400MHz, CDCl3): δ3.78(t,HOCH2CH2CH2CH2CH2CH2NCH3),3.60(t,HOCH2CH2CH2NCH3),2.58(t,HOCH2CH2CH2NCH3),2.36(t,HOCH2CH2CH2CH2CH 2CH2NCH3),2.22(t,HOCH2CH2CH2NCH3),1.68(m,HOCH2CH2CH2NCH3),1.54(m,HOCH2CH2CH2CH2CH2CH2NCH3),1.34(m,HOCH2CH2CH2CH2CH2CH2NCH3).

[0395] HRMS(ESI,m / z):C 10 H 24 NO 2； [M+H] + Calculated value: 190.1807 ； Measured value: 190.1811.

[0396] Preparation Example A15: 7-((3-hydroxypropyl)(methyl)amino)hepta-1-ol (A088)

[0397] Weigh 3-(aminomethyl)-1-propanol (3.00 g, 33.65 mmol), 7-bromo-1-heptanol (11.16 g, 57.21 mmol), and anhydrous potassium carbonate (9.49 g, 68.66 mmol), then add dry acetonitrile (90 mL) and react at 85 °C for 2 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A088 (4.7 g) was obtained with a yield of 68.7%.

[0398] 1 H NMR (400MHz, CDCl3): δ3.78(t,HOCH2CH2CH2CH2CH2CH2CH2NCH3), 3.61(t,HOCH2CH2CH2NCH3), 2.58(t,HOCH2CH2CH2NCH3), 2.35(t,HOCH2CH2CH2CH2CH2CH 2CH2NCH3), 2.22(t,HOCH2CH2CH2NCH3), 1.69(m,HOCH2CH2CH2NCH3), 1.53(m,HOCH2CH2CH2CH2CH2CH2CH2NCH3), 1.32(m,HOCH2CH2CH2CH2CH2CH2CH2NCH3).

[0399] HRMS(ESI,m / z):C 11 H 26 NO 2； [M+H] + Calculated value: 204.1964 ； Measured value: 204.1969.

[0400] Preparation Example A16: 5-(3-(hydroxymethyl)piperidin-1-yl)pentan-1-ol (A131)

[0401] Weigh out 3-piperidinemethanol (3.50 g, 30.39 mmol), 5-bromo-1-pentanol (7.61 g, 45.58 mmol), and anhydrous potassium carbonate (8.19 g, 59.26 mmol), then add 100 mL of dry acetonitrile and react at 85 °C for 2 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A131 (3.37 g) was obtained in 55.1% yield.

[0402] 1H NMR (400MHz, CDCl3): δ3.61(t,CHCH2OH,NCH2CH2CH2CH2CH2OH),3.49(t,CHCH2OH),2.83(d,CHCH2OH),2.63(d,CHCH2OH),2.31(t,NCH2CH2CH2CH2CH2OH),2.1(s,NCH2CH2CH2 CHCH2),1.96(s,NCH2CH2CH2CHCH2),1.55(m,NCH2CH2CH2CHCH2,NCH2CH2CH2CH2CH2OH),1.38(m,NCH2CH2CH2CH2CH2OH),1.2(s,NCH2CH2CH2CHCH2),1.09(m,NCH2CH2CH2CH).

[0403] HRMS(ESI,m / z):C 11 H 24 NO 2； [M+H] + Calculated value: 202.1807 ； Measured value: 202.1810.

[0404] Preparation Example A17: 4-(Ethyl(2-hydroxyethyl)amino)but-1-ol (A089)

[0405] Weigh 2.00 g (17.07 mmol) of 4-ethylamino-1-butanol, 4.27 g (34.13 mmol) of 2-bromoethanol, and 5.66 g (40.96 mmol) of anhydrous potassium carbonate, then add 60 mL of dry acetonitrile and react at 85 °C for 7 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A089 (1.14 g) was obtained in a yield of 41.5%.

[0406] 1 H NMR (400MHz, CDCl3): δ3.63(t,HOCH2CH2CH2CH2NCH2CH3), 3.56(t,HOCH2CH2NCH2CH3), 2.58(t,HOCH2CH2CH2CH 2NCH2CH3), 2.44(t,HOCH2CH2NCH2CH3), 1.65(m,HOCH2CH2CH2CH2NCH2CH3), 1.06(t,HOCH2CH2CH2CH2NCH2CH3).

[0407] HRMS(ESI,m / z):C8H 20 NO 2；[M+H] + Calculated value: 162.1494 ； Measured value: 162.1498.

[0408] Preparation Example A18: 6-((4-hydroxybutyl)(isopropyl)amino)hexane-1-ol (A105)

[0409] Weigh 2.500 g (19.05 mmol) of 4-(isopropylamino)butanol, 17.25 g (95.26 mmol) of 6-bromo-1-hexanol, and 15.80 g (114.32 mmol) of anhydrous potassium carbonate. Then add 100 mL of dry acetonitrile and react at 85 °C for 21.5 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A105 (3.57 g) was obtained in 81.0% yield.

[0410] 1 H NMR(400 Hz, CDCl3): δ3.65(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),3.14(t,HOCH2C H2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),2.48(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2) CH2CH2CH2CH2OH),1.57(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.40(m,HO CH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.06(s,HOCH2CH2CH2CH2N(CH(CH3)2)).

[0411] HRMS(ESI,m / z):C 13 H 30 NO 2； [M+H] + Calculated value: 232.2277 ； Measured value: 232.2281.

[0412] Preparation Example A19: 7-((4-hydroxybutyl)(isopropyl)amino)hepta-1-ol (A106)

[0413] Weigh 2.500 g (19.05 mmol) of 4-(isopropylamino)butanol, 18.59 g (95.26 mmol) of 7-bromo-1-heptanol, and 15.80 g (114.32 mmol) of anhydrous potassium carbonate, then add 100 mL of dry acetonitrile and react at 85 °C for 24 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A106 (3.8 g) was obtained in 81.2% yield.

[0414] 1 H NMR(400MHz, CDCl3): δ3.55(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH(CH3)2)),3.40(t,N(CH(CH3)2)CH2 CH2CH2CH2OH),3.1(m,N(CH(CH3)2)CH2CH2CH2CH2OH),2.44(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH(CH3 )2)CH2CH2CH2CH2OH),1.56(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.36(m, HOCH2CH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.03(s,HOCH2CH2CH2CH2N(CH(CH3)2)).

[0415] HRMS(ESI,m / z):C 14 H 32 NO 2； [M+H] + Calculated value: 246.2433 ； Measured value: 246.2438.

[0416] Preparation Example A20: 5-((2-hydroxyethyl)(isopropyl)amino)pentan-1-ol (A107)

[0417] Weigh 1.000 g (6.89 mmol) of 5-(isopropylamino)pentanol, 4.32 g (34.43 mmol) of 2-bromoethanol, and 5.71 g (41.31 mmol) of anhydrous potassium carbonate, then add 40 mL of dry acetonitrile and react at 85 °C for 26 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A107 (1.03 g) was obtained in 78.6% yield.

[0418] 1 H NMR (400MHz, CDCl3): δ3.62(t,HOCH2CH2CH2CH2CH2N(CH(CH3)2)),3.48(t,HOCH2CH2N(CH(CH3)2 )),2.96(m,HOCH2CH2N(CH(CH3)2)),2.52(t,HOCH2CH2N(CH(CH3)2)),2.40(t,HOCH2CH2CH2CH2CH 2N(CH(CH3)2))),1.56(m,HOCH2CH2CH2CH2CH2N(CH(CH3)2)),1.43(m,HOCH2CH2CH2CH2CH2N(CH(C H3)2))),1.36(m,HOCH2CH2CH2CH2CH2N(CH(CH3)2)),0.99(m,HOCH2CH2CH2CH2CH2N(CH(CH3)2)).

[0419] HRMS(ESI,m / z):C 10 H 24 NO 2； [M+H] + Calculated value: 190.1807 ； Measured value: 190.1811.

[0420] Preparation Example A21: 3,3'-(piperazine-1,4-diyl)bis(propane-1-ol) (A002)

[0421] Anhydrous piperazine (5.00 g, 58.05 mmol) and 3-bromo-1-propanol (28236.88 mg, 203.16 mmol) were dissolved in dry acetonitrile (250 mL), followed by the addition of anhydrous potassium carbonate (3.37 g, 243.79 mmol). The reaction was carried out at 85 °C for 1 h. After the reaction was completed, the potassium carbonate was removed by hot filtration with diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was recrystallized from methanol to give 3,3'-(piperazine-1,4-diyl)bis(propan-1-ol) (A002) (1.1115 g), with a yield of approximately 9.5%.

[0422] 1 H NMR (400MHz, CDCl3): δ5.08(s,HOCH2CH2CH2N),3.77(d,HOCH2CH2CH2N),3.22-2.08(m,HOCH2CH2CH2N(CH2CH2)CH2CH2N),1.70(m,HOCH2CH2CH2N).

[0423] HRMS(ESI,m / z):C10 H 22 N2O 2； [M+H] + Calculated value: 203.1760; Measured value: 203.1761.

[0424] Preparation Example A22: 3-(4-(2-hydroxyethyl)piperidin-1-yl)prop-1-ol (A070)

[0425] 4-Piperidine ethanol (5.00 g, 38.70 mmol) and 3-bromo-1-propanol (1.08 g, 77.40 mmol) were dissolved in acetonitrile (150 mL), and then anhydrous potassium carbonate (1.28 g, 92.88 mmol) was added. The reaction was carried out at room temperature for 13 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in deionized water (30 mL) and then washed with dichloromethane (3 mL). The aqueous layer was collected, lyophilized, and then subjected to C24-dichloromethane treatment. 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A070 (2.7 g) was obtained in a yield of 37.25%.

[0426] 1 H NMR (400MHz, CDCl3): δ3.77(t,HOCH2CH2CH2N),3.65(t,HOCH2CH2CH),3.01(d,HOCH2CH2CH2N),2.62-2.50(m,HOCH2CH2CH2N(CH2)CH2),1 .90(t,HOCH2CH2CH2N),1.69(dd,HOCH2CH2CH2N(CH2CH2)CH2CH2),1.47(dq,HOCH2CH2CH(CH2)CH2),1.28-1.12(m,HOCH2CH2CH(CH2)CH2).

[0427] HRMS(ESI,m / z):C 10 H 21 NO 2； [M+H] + Calculated value: 188.1651; Measured value: 188.1653.

[0428] Preparation Example A23: 3,3'-(1,4-diazacycloheptane-1,4-diyl)bis(prop-1-ol) (A073)

[0429] Peripterazine (1.00 g, 9.98 mmol) and 3-bromo-1-propanol (4.16 g, 29.95 mmol) were dissolved in acetonitrile (50 mL), and then anhydrous potassium carbonate (4.97 g, 35.94 mmol) was added. The reaction was carried out at 85 °C for 5 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A073 (1.16 g) was obtained in a yield of 26.86%.

[0430] 1 H NMR (400MHz, CDCl3): δ4.55(s,HOCH2CH2CH2),3.80(t,HOCH2CH2CH2),2.80-2.62(m,HOCH2CH2CH2N(CH2 CH2)CH2CH2CH2N2CH2CH2CH2OH),1.82(m,HOCH2CH2CH2N(CH2CH2)CH2CH2CH2N),1.66(m,HOCH2CH2CH2).

[0431] HRMS(ESI,m / z):C 11 H 24 N2O 2； [M+H] + Calculated value: 217.1916 ； Measured value: 217.1917.

[0432] Preparation Example A24: 3-(3-(hydroxymethyl)pyrrolidone-1-yl)prop-1-ol (A074)

[0433] Pyridinyl-3-ylmethanol (0.9010 g, 8.90 mmol) and 3-bromo-1-propanol (2.47 g, 17.80 mmol) were dissolved in dry acetonitrile (150 mL), followed by the addition of anhydrous potassium carbonate (2.95 g, 21.35 mmol), and the reaction was carried out at 85 °C for 2 h. After the reaction was complete, the anhydrous potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in deionized water (5 mL) and washed three times with dichloromethane (7 mL). The aqueous layer was collected, lyophilized, and then subjected to C24-445 ... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A074 (0.57 g) was obtained, with a yield of 40.1%.

[0434] 1H NMR (400MHz, CDCl3): δ3.77(t,HOCH2CH2CH2N),3.54(dd,HOCH2CH(CH2)CH2CH2),3.47(dd,HOCH2CH(CH2)CH2CH2),2.79-2.29(m,HOCH2 CH(CH2)CH2CH2NCH2CH2),1.94(m,HOCH2CH(CH2)CH2CH2NCH2CH2),1.71(dd,HOCH2CH2CH2N),1.49(td,HOCH2CH(CH2)CH2CH2NCH2CH2).

[0435] HRMS(ESI,m / z):C8H 17 NO 2； [M+H] + Calculated value: 160.1338 ； Measured value: 160.1339.

[0436] Preparation Example A25: 2,2'-(piperidine-1,2-diyl)bis(ethanol) (A071)

[0437] 2-Piperidinol (6.00 g, 46.44 mmol) and 2-bromoethanol (10.36 g, 83.59 mmol) were dissolved in dry acetonitrile (120 mL), and then anhydrous potassium carbonate (1.39 mg, 100.31 mmol) was added. The reaction was carried out at 85 °C for 8 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was analyzed by C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A071 (6.2 g) was obtained, with a yield of 77.0%.

[0438] 1 H NMR (400MHz, CDCl3): δ3.79(t,NCHCH2CH2OH),3.55(t,NCH2CH2OH),2.45-2.28(m,CH2N(CH)CH2),1.58-1.47(m,CH2CH2CH2CH(CH2)NCH2CH2OH).

[0439] HRMS(ESI,m / z): C9H 19 NO 2； [M+H] + Calculated value: 174.1494 ； Measured value: 174.1484.

[0440] Preparation Example A26: 3-(2-(2-hydroxyethyl)piperidin-1-yl)prop-1-ol (A082)

[0441] 2-Piperidine ethanol (6.00 g, 46.44 mmol) and 3-bromo-1-propanol (11.6 g, 83.59 mmol) were dissolved in dry acetonitrile (120 mL), and then anhydrous potassium carbonate (13.9 g, 100.31 mmol) was added. The reaction was carried out at 85 °C for 8 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was analyzed by C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A082 (5.8 g) was obtained, with a yield of 66.7%.

[0442] 1 H NMR (400MHz, CDCl3): δ3.78(t,NCHCH2CH2OH),3.54(t,NCH2CH2CH2OH),2.45-2.28(m,CH2N(CH)CH2),1.58-1.47(m,CH2CH2CH2CH(CH2)NCH2CH2CH2OH).

[0443] HRMS(ESI,m / z): C 10 H 21 NO 2； [M+H] + Calculated value: 188.1651 ； Measured value: 188.1654.

[0444] Preparation Example A27: 3-(3-(hydroxymethyl)piperidin-1-yl)propanol (A075)

[0445] Weigh 1.000 g (8.680 mmol) of 3-piperidinemethanol, 2.052 g (14.76 mmol) of 3-bromo-1-propanol, and 2.46 g (17.71 mmol) of anhydrous potassium carbonate, then add 30 mL of dry acetonitrile and react at 85 °C for 13 h. After the reaction solution returns to room temperature, filter. Collect the organic layer and concentrate under reduced pressure to obtain the crude product. The crude product is then subjected to C... 18 After column-column purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A0075 (0.93 g) was obtained in a yield of 62%.

[0446] 1H NMR (400MHz, CDCl3): δ3.77(t,CHCH2OH),3.54-3.42(m,NCH2CH2CH2OH),2.98(d,CHCH2OH),2.8 3(s,CHCH2OH),2.57(t,NCH2CH2CH2CH),2.01(m,NCH2CH2CH2OH),1.91-1.64(m,NCH2CH2CH2CHCH 2, NCH2CH2CH2OH),1.52(m,NCH2CH2CH2CHCH2),1.06(m,NCH2CH2CH2CH).

[0447] HRMS(ESI,m / z):C9H 20 NO2; [M+H] + Calculated value: 174.1494 ； Measured value: 174.1498

[0448] Preparation Example A28: 2,2'-(piperidine-1,4-diyl)bis(1-ethanol) (A078)

[0449] 4-Piperidinol (5.00 g, 38.70 mmol) and 2-bromoethanol (1.21 g, 96.75 mmol) were dissolved in dry acetonitrile (250 mL), followed by the addition of anhydrous potassium carbonate (1.60 g, 116.10 mmol), and the reaction was carried out at 85 °C for 2 h. After the reaction was complete, the mixture was allowed to return to room temperature, and the anhydrous potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in deionized water (40 mL) and then washed twice with dichloromethane (40 mL). The aqueous layer was collected, lyophilized, and then subjected to C24-dichloromethane treatment. 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A078 (2.25 g) was obtained, with a yield of 33.5%.

[0450] 1 H NMR (400MHz, CDCl3): δ3.69(t,HOCH2CH2CH),3.59(t,HOCH2CH2N),2.89(d,HOCH2CH2CH(CH2CH2)CH2CH2),2.54–2.47(m,HOCH2CH2N),2.04(td,HOCH2 CH2CH(CH2CH2)CH2CH2),1.70(d,HOCH2CH2N(CH2CH2)CH2CH2),1.52(q,HOCH2CH2CH),1.45(dd,HOCH2CH2CH),1.25(qd,HOCH2CH2CH(CH2CH2)CH2CH2).

[0451] HRMS(ESI,m / z):C9H 19 NO 2； [M+H] + Calculated value: 174.1494 ； Measured value: 174.1495.

[0452] Preparation Example A29: 3-((2-hydroxyethyl)(isopropyl)amino)prop-1-ol (A095)

[0453] 3-(isopropylamino)propanol (2.50 g, 21.33 mmol) and 2-bromoethanol (18.66 g, 149.33 mmol) were dissolved in dry acetonitrile (250 mL), and then anhydrous potassium carbonate (2.48 g, 179.19 mmol) was added. The reaction was carried out at 85 °C for 12 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A095 (2.5 g) was obtained, with a yield of 72.7%.

[0454] 1 H NMR (400MHz, CDCl3): δ3.79(t,HOCH2CH2CH2NCH2CH2OH),3.62(t,HOCH2CH2CH2NCH2CH2OH),3.07-2.97(m,HOCH2CH2CH2N(CH(CH3)2)CH2CH2OH),2.66(t,HOCH2CH2CH 2N(CH(CH3)2)CH2CH2OH),2.56(t,HOCH2CH2CH2N(CH(CH3)2)CH2CH2OH),1.68(m,HOCH2CH2CH2N(CH(CH3)2)CH2CH2OH),1.01(d,HOCH2CH2CH2N(CH(CH3)2)CH2CH2OH).

[0455] HRMS(ESI,m / z):C8H 19 NO 2； [M+H] + Calculated value: 162.1494 ； Measured value: 162.1499.

[0456] Preparation Example A30: 3,3'-(isopropylazadiyl)bis(propan-1-ol) (A096)

[0457] The procedure was the same as described in Preparation Example A29, except that 2-bromoethanol was replaced with 3-bromo-1-propanol, and the reaction time was 24 h. After C... 18After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A096 (3.85 g) was obtained, with a yield of 85.7%.

[0458] 1 H NMR (400MHz, CDCl3): δ3.77-3.60(m,HOCH2CH2CH2N(CH(CH3)2)CH2CH2CH2OH), 3.06(m,HOCH2CH2CH2N(CH(CH3)2)CH2CH2CH2OH), 2.61-2.39(m,HO CH2CH2CH2N(CH(CH3)2)CH2CH2CH2OH),1.64-1.53(m,HOCH2CH2CH2N(CH(CH3)2)CH2CH2CH2OH),0.99(d,HOCH2CH2CH2N(CH(CH3)2)CH2CH2CH2OH).

[0459] HRMS(ESI,m / z):C9H 21 NO2; [M+H] + Calculated value: 176.1651; Measured value: 176.1655.

[0460] Preparation Example A31: 5-((3-hydroxypropyl)(isopropyl)amino)pentan-1-ol (A098)

[0461] The process is the same as described in Preparation Example A29, except that 2-bromoethanol is replaced with 5-bromo-1-pentanol. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A098 (3.82 g) was obtained, with a yield of 88.0%.

[0462] 1 H NMR (400MHz, CDCl3): δ3.79(t,HOCH2CH2CH2NCH2CH2CH2CH2CH2OH),3.62(t,HOCH2CH2CH2NCH2CH2CH2CH2CH2OH),3.07-2.97(m,HOCH2CH2CH2N(CH(CH3)2)CH2),2.64(t,HOCH 2CH2CH2NCH2CH2),2.56(t,HOCH2CH2CH2NCH2),1.68(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2OH),1.39(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2OH),1.01(d,HOCH2CH2CH2NCH(CH3)2).

[0463] HRMS(ESI,m / z):C 11H 25 NO 2； [M+H] + Calculated value: 204.1964 ； Measured value: 204.1967.

[0464] Preparation Example A32: 6-((3-hydroxypropyl)(isopropyl)amino)hexane-1-ol (A099)

[0465] The procedure was the same as described in Preparation Example A29, except that 2-bromoethanol was replaced with 6-bromo-1-hexanol, and the reaction time was 20 h. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A099 (2.05 g) was obtained, with a yield of 66.6%.

[0466] 1 H NMR (400MHz, CDCl3): δ3.78(t,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),3.62(t,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),3.08-2.93(m,HOCH2CH2CH2N(CH(CH3)2)CH2),2.64(t,HOCH 2CH2CH2NCH2CH2),2.56(t,HOCH2CH2CH2NCH2),1.68(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),1.37(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),1.03(d,HOCH2CH2CH2NCH(CH3)2).

[0467] HRMS(ESI,m / z):C 12 H 27 NO 2； [M+H] + Calculated value: 218.2120 ； Measured value: 218.2122.

[0468] Preparation Example A33: 7-((3-hydroxypropyl)(isopropyl)amino)heptane-1-ol (A100)

[0469] The process is the same as described in Preparation Example A29, except that 2-bromoethanol is replaced with 7-bromo-1-heptanol. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A100 (3.89 g) was obtained, with a yield of 78.7%.

[0470] 1H NMR (400MHz, CDCl3): δ3.78(t,HOCH2CH2CH2NCH2CH2CH2CH2),3.62(t,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),3.08-2.93(m,HOCH2CH2CH2N(CH(CH3)2)CH2),2.64(t,HOCH2CH2C H2NCH2CH2),2.56(t,HOCH2CH2CH2NCH2),1.68(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.38(m,HOCH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.02(d,HOCH2CH2CH2NCH(CH3)2).

[0471] HRMS(ESI,m / z):C 13 H 29 NO 2； [M+H] + Calculated value: 232.2277 ； Measured value: 232.2279.

[0472] Preparation Example A34: 5-(Butyl(4-hydroxybutyl)amino)pentan-1-ol (A116)

[0473] 4-(butanoamino)-1-butanol (2.00 g, 13.77 mmol) and 5-bromo-1-pentanol (11.52 g, 68.85 mmol) were dissolved in acetonitrile (200 mL), and then anhydrous potassium carbonate (1.14 g, 82.62 mmol) was added. The reaction was carried out at 85 °C for 8 h. After the reaction was completed, the potassium carbonate was removed by filtration through diatomaceous earth moistened with tetrahydrofuran. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A116 (2.53 g) was obtained, with a yield of 79.3%.

[0474] 1H NMR(400MHz,CDCl3):3.64(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2OH),3.57(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),2.54(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3) ,1.87(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2OH),1.69-1.32(m,HOCH2CH2CH2CH2N(CH2CH2CH2CH3)CH2CH2CH2CH2CH2OH).,0.93(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3)

[0475] HRMS(ESI,m / z):C 13 H 29 NO 2； [M+H] + Calculated value: 232.2277 ； Measured value: 232.2279.

[0476] Preparation Example A35: 6-(Butyl(4-hydroxybutyl)amino)hexane-1-ol (A117)

[0477] The procedure was the same as described in Preparation Example A34, except that 5-bromo-1-pentanol was replaced with 6-bromo-1-hexanol, and the reaction time was 14 h. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A117 (2.78 g) was obtained, with a yield of 65.9%.

[0478] 1 H NMR (400MHz, CDCl3): δ3.65(d,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),3.41(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),2.54(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3) 1.87(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2OH),1.69-1.32(m,HOCH2CH2CH2CH2N(CH2CH2CH2CH3)CH2CH2CH2CH2CH2CH2OH).,0.93(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3)

[0479] HRMS(ESI,m / z):C 14 H37 NO 2； [M+H] + Calculated value: 246.2433 ； Measured value: 246.2436.

[0480] Preparation Example A36: 7-(Butyl(4-hydroxybutyl)amino)heptane-1-ol (A118)

[0481] The procedure was the same as described in Preparation Example A34, except that 5-bromo-1-pentanol was replaced with 7-bromo-1-heptanol, and the reaction time was 8 hours. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A118 (3.14 g) was obtained, with a yield of 70.4%.

[0482] 1 H NMR (400MHz, CDCl3): δ3.64(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),3.40(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),2.54(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3) 1.86(m,HOCH2CH2CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.56-1.36(m,HOCH2CH2CH2CH2N(CH2CH2CH2CH3)CH2CH2CH2CH2CH2CH2CH2OH),0.95(t,HOCH2CH2CH2CH2NCH2CH2CH2CH3)

[0483] HRMS(ESI,m / z):C 15 H 33 NO 2； [M+H] + Calculated value: 260.2590 ； Measured value: 260.2594.

[0484] Preparation Example A37: 5,5'-(piperazine-1,4-diyl)bis(1-pentanol) (A119)

[0485] The process is the same as described in Preparation Example A21, except that 3-bromo-1-propanol is replaced with 5-bromo-1-pentanol. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A119 (2.65 g) was obtained, with a yield of 44.2%.

[0486] 1H NMR (400MHz, CDCl3): δ3.62(t,HOCH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),2.53-2.16(m,HOCH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2 CH2CH2OH),1.55(m,HOCH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),1.46-1.32(m,HOCH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH).

[0487] HRMS(ESI,m / z):C 14 H 30 N2O 2； [M+H] + Calculated value: 259.2386 ； Measured value: 259.2391.

[0488] Preparation Example A38: 6,6'-(piperazine-1,4-diyl)bis(hexane-1-ol) (A120)

[0489] The process is the same as described in Preparation Example A21, except that 3-bromo-1-propanol is replaced with 6-bromo-1-hexanol. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A120 (5.55 g) was obtained, with a yield of 83.5%.

[0490] 1 H NMR (400MHz, CDCl3): δ3.63(t,HOCH2(CH2)4CH2N(CH2CH2)CH2CH2NCH2(CH2)4CH2OH), 3.00-1.96( m,HOCH2(CH2)4CH2N(CH2CH2)CH2CH2NCH2(CH2)4CH2OH),1.61-1.53(m,HOCH2CH2(CH2)3CH2N(CH2 CH2)CH2CH2NCH2(CH2)3CH2CH2OH),1.53-1.45(m,HOCH2CH2CH2(CH2)2CH2N(CH2CH2)CH2CH2NCH2( CH2)2CH2CH2CH2OH),1.35(m,HOCH2CH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2OH).

[0491] HRMS(ESI,m / z):C 16 H 34 N2O 2； [M+H] + Calculated value: 287.2699 ； Measured value: 287.2702.

[0492] Preparation Example A39: 7,7'-(piperazine-1,4-diyl)bis(heptane-1-ol) (A121)

[0493] The process is the same as described in Preparation Example A21, except that 3-bromo-1-propanol is replaced with 7-bromo-1-heptanol. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A121 (3.63 g) was obtained, with a yield of 49.7%.

[0494] 1 H NMR (400MHz, CDCl3): δ3.63(td,HOCH2(CH2)5CH2N(CH2CH2)CH2CH2NCH2(CH2)5CH2OH),2.93-2.21(m,HOCH2(CH2)5CH2N(CH2C H2)CH2CH2NCH2(CH2)5CH2OH),1.86(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.56(dd, HOCH2CH2CH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.50-1.41(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH2C H2)CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.35(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH).

[0495] HRMS(ESI,m / z):C 18 H 38 N2O 2； [M+H] + Calculated value: 315.3012 ； Measured value: 315.3016.

[0496] Preparation Example A40: 5,5'-(1,4-diazacycloheptane-1,4-diyl)bis(1-pentan-1-ol) (A122)

[0497] The procedure was the same as described in Preparation Example A21, except that anhydrous piperazine was replaced with perpiperazine, 3-bromo-1-propanol was replaced with 5-bromo-1-pentanol, the molar ratio of perpiperazine / 5-bromo-1-pentanol / potassium carbonate was 1 / 3.5 / 4.2, and the reaction time was 2 hours. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A122 (3.45 g) was obtained, with a yield of 63.4%.

[0498] 1 H NMR (400MHz, CDCl3): δ3.62(t,HOCH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),2.68(d,HOCH2CH2CH2 CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),2.53-2.41(m,HOCH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2C H2CH2CH2OH),1.79(p,HOCH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),1.53(dq,HOCH2CH2CH2CH2CH2 N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH),1.39(q,HOCH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2OH).

[0499] HRMS(ESI,m / z):C 15 H 32 N2O 2； [M+H] + Calculated value: 273.2542 ； Measured value: 273.2546.

[0500] Preparation Example A41: 6,6'-(1,4-diazacycloheptane-1,4-diyl)bis(hexane-1-ol) (A123)

[0501] The process was the same as described in Preparation Example A21, except that anhydrous piperazine was replaced with perpiperazine, 3-bromo-1-propanol was replaced with 6-bromo-1-hexanol, the molar ratio of perpiperazine / 6-bromo-1-hexanol / potassium carbonate was 1 / 3.5 / 4.2, and the reaction time was 2 hours. After C... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A123 (4.37 g) was obtained, with a yield of 72.8%.

[0502] 1 H NMR (400MHz, CDCl3): δ3.61(t,HOCH2(CH2)4CH2N(CH2CH2CH2)CH2CH2NCH2(CH2)4CH2OH),2.81-2.63(m,HOCH2(CH2)4CH2N(CH2CH2CH2)CH2 CH2NCH2(CH2)4CH2OH),2.48-2.40(m,HOCH2(CH2)4CH2N(CH2CH2CH2)CH2CH2NCH2(CH2)4CH2OH),1.82-1.73(m,HOCH2(CH2)4CH2N(CH2CH2CH 2)CH2CH2NCH2(CH2)4CH2OH),1.56(m,HOCH2CH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2OH),1.47(m,HOCH2CH2CH2CH2CH2 CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2OH),1.35-1.22(m,HOCH2CH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2OH).

[0503] HRMS(ESI,m / z):C 17 H 36 N2O 2； [M+H] + Calculated value: 301.2855 ； Measured value: 301.2859.

[0504] Preparation Example A42: 7,7'-(1,4-diazacycloheptane-1,4-diyl)bis(heptane-1-ol) (A124)

[0505] The procedure was the same as described in Preparation Example A21, except that anhydrous piperazine was replaced with perpiperazine, 3-bromo-1-propanol was replaced with 7-bromo-1-heptanol, the molar ratio of perpiperazine / 7-bromo-1-heptanol / potassium carbonate was 1 / 3.5 / 4.2, and the reaction time was 1.5 h. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A124 (4.43 g) was obtained, with a yield of 67.5%.

[0506] 1H NMR (400MHz, CDCl3): δ3.63(t,HOCH2(CH2)5CH2N(CH2CH2CH2)CH2CH2NCH2(CH2)5CH2OH), 3.42-3.34(m, HOCH2(CH2)5CH2NCH2CH2C H2NCH2(CH2)5CH2OH),2,68-2.44(m,HOCH2(CH2)5CH2NCH2CH2NCH2(CH2)5CH2OH),1.88-1.78(m,HOCH2(CH2)5CH2NCH2CH2CH2NCH2 (CH2)5CH2OH),1.56(m,HOCH2CH2CH2CH2CH2CH2CH2NCH2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.45(m,HOCH2CH2CH2CH2CH2CH2CH2NCH 2CH2NCH2CH2CH2CH2CH2CH2CH2OH),1.35-1.24(m,HOCH2CH2CH2CH2CH2CH2CH2N(CH2CH2CH2)CH2CH2NCH2CH2CH2CH2CH2CH2CH2OH).

[0507] HRMS(ESI,m / z):C 19 H 40 N2O 2； [M+H] + Calculated value: 329.3168 ； Measured value: 329.3171.

[0508] Preparation Example A43: 5-(2-(2-hydroxyethyl)piperidin-1-yl)pentan-1-ol (A128)

[0509] The procedure was the same as described in Preparation Example A21, except that anhydrous piperazine was replaced with 2-piperidineethanol, 3-bromo-1-propanol was replaced with 5-bromo-1-pentanol, the molar ratio of 2-piperidineethanol / 5-bromo-1-pentanol / potassium carbonate was 1 / 2.5 / 3, and the reaction time was 10 h. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A128 (2.08 g) was obtained, with a yield of 62.5%.

[0510] 1H NMR (400MHz, CDCl3): δ3.78(t,NCHCH2CH2OH),3.54(t,NCH2CH2CH2CH2CH2OH),2.45-2.28(m,CH2N(CH)CH2CH2CH2),1.58-1.47(m,CH2CH2CH2CH(CH2)NCH2CH2CH2CH2CH2OH).

[0511] HRMS(ESI,m / z):C 12 H 25 NO 2； [M+H] + Calculated value: 216.1964 ； Measured value: 216.1967.

[0512] Preparation Example A44: 6-(2-(2-hydroxyethyl)piperidin-1-yl)hexane-1-ol (A129)

[0513] The procedure was the same as described in Preparation Example A21, except that anhydrous piperazine was replaced with 2-piperidineethanol, 3-bromo-1-propanol was replaced with 6-bromo-1-hexanol, the molar ratio of 2-piperidineethanol / 6-bromo-1-hexanol / potassium carbonate was 1 / 2.5 / 3, and the reaction time was 10 h. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A129 (2.30 g) was obtained, with a yield of 64.8%.

[0514] 1 H NMR (400MHz, CDCl3): δ3.77(t,NCHCH2CH2OH),3.52(t,NCH2CH2CH2CH2CH2CH2OH),2.45-2.28(m,CH2N(CH)CH2CH2CH2),1.56-1.42(m,CH2CH2CH2CH(CH2)NCH2CH2CH2CH2CH2CH2OH).

[0515] HRMS(ESI,m / z):C 13 H 27 NO 2； [M+H] + Calculated value: 230.2120 ； Measured value: 230.2124.

[0516] Preparation Example A45: 7-(2-(2-hydroxyethyl)piperidin-1-yl)heptane-1-ol (A130)

[0517] The procedure was followed as described in Preparation Example A21, except that anhydrous piperazine was replaced with 2-piperidineethanol, 3-bromo-1-propanol was replaced with 7-bromo-1-heptanol, the molar ratio of 2-piperidineethanol / 7-bromo-1-heptanol / potassium carbonate was 1 / 2.5 / 3, and the reaction time was 10 h. After C... 18 After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A130 (2.51 g) was obtained, with a yield of 67.7%.

[0518] 1 H NMR (400MHz, CDCl3): δ3.79(t,NCHCH2CH2OH),3.53(t,NCH2CH2CH2CH2CH2CH2CH2OH),2.45-2 .28(m,CH2N(CH)CH2CH2CH2),1.57-1.43(m,CH2CH2CH2CH(CH2)NCH2CH2CH2CH2CH2CH2CH2OH).

[0519] HRMS(ESI,m / z):C 14 H 29 NO 2； [M+H] + Calculated value: 244.2277 ； Measured value: 244.2280.

[0520] Preparation Example A46: 6-(3-(hydroxymethyl)piperidin-1-yl)hex-1-ol (A132)

[0521] 3-Piperidinemethanol (4.000 g, 34.73 mmol), 6-bromo-1-hexanol (10.70 g, 59.04 mmol), and anhydrous potassium carbonate (10.61 g, 76.76 mmol) were weighed out, and then 120 mL of dry acetonitrile was added. The reaction mixture was reacted at 85 °C for 4.5 h. After the reaction solution returned to room temperature, it was filtered. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting with acetonitrile and 0.5% ammonia) to obtain monomer A132 (4.45 g), with a yield of 59.5%.

[0522] 1H NMR (400MHz, CDCl3): δ3.62(m,CHCH2OH,NCH2CH2CH2CH2CH2CH2OH),3.51(m,CHCH2OH),2.80(d,CHCH2OH),2.58(d,CHCH2OH),2.29(t,NCH2CH2CH2CH2CH2C H2OH),2.15(s,NCH2CH2CH2CHCH2),2.02(s,NCH2CH2CH2CHCH2),1.79(t,NCH2CH2CH2CH2CH2CH2OH),1.68(m,NCH2CH2CH2CHCH2),1.55(m,NCH2CH2CH2CHCH 2, NCH2CH2CH2CH2CH2CH2OH),1.36(m,NCH2CH2CH2CH2CH2CH2OH),1.09(m,NCH2CH2CH2CH).

[0523] HRMS(ESI,m / z):C 12 H 26 NO 2； [M+H] + Calculated value: 216.1964 ； Measured value: 216.1968.

[0524] Preparation Example A47: 7-(3-(hydroxymethyl)piperidin-1-yl)hepta-1-ol (A133)

[0525] 3-Piperidinemethanol (3.500 g, 30.39 mmol), 7-bromo-1-heptanol (8.90 g, 45.58 mmol), and anhydrous potassium carbonate (8.19 g, 59.26 mmol) were weighed out, and then 100 mL of dry acetonitrile was added. The reaction mixture was reacted at 85 °C for 3 h. After the reaction solution returned to room temperature, it was filtered. The organic layer was collected and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting with acetonitrile and 0.5% ammonia) to obtain monomer A133 (3.35 g), with a yield of 48.1%.

[0526] 1 H NMR (400MHz, CDCl3): δ3.62-3.49(m,CHCH2OH,NCH2CH2CH2CH2CH2CH2CH2OH),2.83(d,CHCH2OH),2.62(d,CHCH2OH),2.28 (t,NCH2CH2CH2CH2CH2CH2CH2OH),2.1-1.97(m,NCH2CH2CH2CHCH2),1.75(m,NCH2CH2CH2CHCH2),1.54(m,NCH2CH2CH2CHCH2, NCH2CH2CH2CH2CH2CH2CH2OH),1.32(m,NCH2CH2CH2CH2CH2CH2CH2OH),1.09(m,NCH2CH2CH2CH).

[0527] HRMS(ESI,m / z):C 13 H 28 NO 2； [M+H] + Calculated value: 230.2120 ； Measured value: 230.2122.

[0528] Preparation Example B1: (Methylazonyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B003)

[0529] Under ice bath conditions, A003 (N-methyldiethanolamine, 20.0 g, 1 eq) was dissolved in 200 mL of DCM solvent, and CDI (68.1 g, 2.5 eq) was added. The mixture was brought back to room temperature and stirred at room temperature for 1 hour. After the reaction was completed, water was added to quench the reaction. The mixture was extracted three times with 200 mL of DCM, and the organic phases were combined and washed three times with pure water (500 mL × 3). The organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain B003 yellow solid (40 g, 77.6%).

[0530] 1 H NMR (700MHz, CDCl3): δ8.12(t,J=1.1Hz,2H),7.39(t,J=1.5Hz,2H),7.08–7.04(m,2H),4.49(t,J=5.7Hz,4H),2.88(t,J=5.7Hz,4H),2.43(s,3H).

[0531] Preparation Example B2: Piperazine-1,4-diylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylate) (B004)

[0532] Following the procedure of Preparation Example B1, except that A003 was replaced with A004 (1,4-bis(2-hydroxyethyl)piperazine), yielding B004 as a yellow oil with a yield of 96.0%.

[0533] 1 H NMR (700MHz, CDCl3): δ8.14(d,J=1.3Hz,2H),7.43(t,J=1.5Hz,2H),7.14–7.05(m,2H),4.52(t,J=5.8Hz,4H),2.77(t,J=5.8Hz,4H),2.56(s,8H).

[0534] Preparation Example B3: (Ethylazanediyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B005)

[0535] The process was the same as in Preparation Example B1, except that A003 was replaced with A005 (N-ethyldiethanolamine), resulting in B005 as a yellow oil with a yield of 55.0%.

[0536] 1 H NMR (700MHz, CDCl3): δ8.07(t,J=1.1Hz,2H),7.34(t,J=1.5Hz,2H),7.03–7.00(m,2H),4 .41(t,J=5.9Hz,4H),2.89(t,J=5.9Hz,4H),2.65(t,J=7.1Hz,2H),1.02(t,J=7.1Hz,3H).

[0537] Preparation Example B4: (Butylazanediyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B007)

[0538] Following the procedure of Preparation Example B1, except that A003 was replaced with A007 (N-butyldiethanolamine), resulting in B007 as a yellow oil with a yield of 73.7%.

[0539] 1 H NMR (700MHz, CDCl3): δ8.09(t,J=1.1Hz,2H),7.36(t,J=1.5Hz,2H),7.04(q,J=2.0,1.3Hz,2H),4.43(t,J=5.9Hz,4H), 2.91(t,J=5.9Hz,4H),2.58–2.54(m,2H),1.39(ddt,J=9.0,7.5,3.6Hz,2H),1.28–1.24(m,2H),0.85(t,J=7.3Hz,3H).

[0540] Preparation Example B5: 3-(1-(2-((1H-imidazol-1-carbonyl)oxy)ethyl)piperidin-4-yl)propyl 1H-imidazol-1-carboxylic acid ester (B021)

[0541] Following the procedure of Preparation Example B1, except that A003 was replaced with A021(3-(1-(2-hydroxyethyl)piperidin-4-yl)prop-1-ol, yielding a yellow solid B021 in 66.8% yield.

[0542] 1H NMR (700MHz, CDCl3): δ8.12(dt,J=4.0,1.1Hz,2H),7.41(dt,J=5.1,1.5Hz,2H),7.08–7.05(m,2H),4.50(t,J=5.8Hz,2H),4.39(t,J= 6.7Hz,2H),2.74(t,J=5.8Hz,2H),2.07(td,J=11.4,2.4Hz,2H),1.82–1.77(m,2H),1.69(dt,J=12.8,2.2Hz,2H),1.37–1.18(m,7H).

[0543] Preparation Example B6: Thiobis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B025)

[0544] The procedure was the same as in Example B1, except that A003 was replaced with A025 (2,2'-thiobis(1-ethanol), yielding B025 as a white solid with a yield of 52.6%).

[0545] 1 H NMR (700MHz, CDCl3): δ8.13(t,J=1.1Hz,2H),7.41(t,J=1.5Hz,2H),7.10–7.06(m,2H),4.57(t,J=6.8Hz,4H),2.98(t,J=6.8Hz,4H).

[0546] Preparation Example B7: Dithiodimethylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B026)

[0547] Following the procedure of Preparation Example B1, except that A003 was replaced with A026 (2,2'-dithionide dibis(1-ethanol), yielding B026 as a white solid in 81.8% yield.

[0548] 1 H NMR (700MHz, CDCl3): δ8.13(t,J=1.1Hz,2H),7.41(t,J=1.5Hz,2H),7.08–7.05(m,2H),4.67(t,J=6.5Hz,4H),3.07(t,J=6.6Hz,4H).

[0549] Preparation Example B8: Trithionylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B027)

[0550] The procedure was the same as in Example B1, except that A003 was replaced with A027 (2,2'-trithionide dimethylbis(1-ethanol), yielding B027 as a white solid in 55.0% yield.

[0551] 1 H NMR (700MHz, CDCl3): δ8.18–8.13(m,2H),7.43(t,J=1.6Hz,2H),7.09–7.05(m,2H),4.73(t,J=6.5Hz,4H),3.26(t,J=6.5Hz,4H).

[0552] Preparation Example B9: (propane-2,2-diylbis(thioalkyldiyl))bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B028)

[0553] Following the procedure of Preparation Example B1, except that A003 was replaced with A028(2,2'-(propane-2,2-diylbis(thioalkyldiyl))bis(ethanol-1-ol), yielding B028 as a yellow oil in 89.4% yield.

[0554] 1 H NMR (700MHz, CDCl3): δ8.15(t,J=1.1Hz,2H),7.43(t,J=1.5Hz,2H),7.08(t,J=1.3Hz,2H),4.57(t,J=7.0Hz,4H),3.02(t,J=7.0Hz,4H),1.67(s,6H).

[0555] Preparation Example B10: (propane-2,2-diylbis(thioalkyldiyl))bis(propane-3,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B029)

[0556] The process was similar to that of Preparation Example B1, except that A003 was replaced with A029(2,2'-(propane-2,2-diylbis(thioalkyldiyl))bis(ethanol-1-ol), resulting in B029 as a yellow oil with a yield of 87%.

[0557] 1 H NMR (700MHz, CDCl3): δ8.15(t,J=1.0Hz,2H),7.43(t,J=1.5Hz,2H),7.12–7.05(m,2 H), 4.50 (t, J = 6.3Hz, 4H), 2.74 (t, J = 7.2Hz, 4H), 2.08 (t, J = 6.7Hz, 4H), 1.61 (s, 6H).

[0558] Preparation Example B11: M008

[0559] 2-Hydroxyethyl disulfide (1 g, 6.48 mmol) and diphenyl carbonate (2.08 g, 9.72 mmol) were dissolved in anhydrous toluene (400 mL), and then Novozymes lipase 435 (N-435) equivalent to 2.08 g of diphenyl carbonate was added. The reaction mixture was reacted at 70 °C under nitrogen for 12 hours, and the lipase was removed by filtration. The collected filtrate was concentrated under reduced pressure to obtain a solid, which was washed with methanol to obtain a crude product. The crude product was recrystallized from ethyl acetate to give white crystals M008, with a yield of approximately 60%.

[0560] 1 H-NMR (700MHz, CDCl3): δ4.37(m,OCOCH2CH2SS), 3.05(m,OCOCH2CH2SS).

[0561] Preparation Example B12: M009

[0562] A003 (0.77 g, 6.48 mmol) and diphenyl carbonate (2.08 g, 9.72 mmol) were dissolved in anhydrous toluene (400 mL), and then 2.08 g of Novozymes lipase 435 (N-435) was added. The reaction mixture was reacted at 70 °C under nitrogen for 12 hours, and the lipase was removed by filtration. The collected filtrate was concentrated under reduced pressure to obtain a solid, which was then separated by silica gel column chromatography (eluent was methanol and ethyl acetate, v / v). The crude product was recrystallized from ethyl acetate to give white crystals M009 in 43% yield.

[0563] 1 H-NMR (700MHz, CDCl3): δ4.2(t,OCOCH2CH2NCH3), 2.69(t,OCOCH2CH2NCH3), 2.34(s,OCOCH2CH2NCH3).

[0564] Example 2: Polymer Preparation

[0565] The monomers used in the polymer preparation examples are summarized in Table 3 below.

[0566] Table 3. Monomers used in polymer preparation examples

[0567] Preparation Example P1: Polymer 244

[0568] Polymer 244 contains the following repeating units:

[0569] Raw materials A003 (185 mg, 1 eq), B025 (502 mg, 1.04 eq), and cesium fluoride (24.7 mg, 0.1 eq) were dissolved in 2 mL of ethyl acetate solution and reacted at 60 °C for 15 hours. After the reaction was completed, the mixture was brought to room temperature, and 2 mL of the reaction solution was precipitated with 50 mL of diethyl ether. The precipitate was centrifuged at 4000 x g for 5 minutes, the supernatant was removed, and the organic phase was evaporated to obtain a pale yellow solid product. 20 mL of acidic aqueous solution (pH = 5) was added for dissolution and ultrafiltration (MWCO: 1000). After ultrafiltration by 10 times the volume, the solution was concentrated to 10 mL, filtered through a 0.2 μm filter, and lyophilized to obtain a fluffy white solid 244 (353 mg, yield 75.7%). Mw: 9.9 kDa.

[0570] Preparation Example P2: Polymer 245

[0571] Polymer 245 contains the following repeating units:

[0572] Following the procedure of Preparation Example P1, the starting materials were A004 (265 mg, 1 eq), B025 (501 mg, 1.06 eq), and cesium fluoride (23.31 mg, 0.1 eq), yielding a fluffy white solid 245 (467 mg, yield 85.5%). Mw: 10.3 kDa.

[0573] Preparation Example P3: Polymer 246

[0574] Polymer 246 contains the following repeating units:

[0575] Following the procedure of Preparation Example P1, the starting materials were A003 (151.9 mg, 1 eq), B027 (507.2 mg, 1.05 eq), and cesium fluoride (19.3 mg, 0.1 eq), yielding a fluffy yellow solid 246 (289 mg, yield 60.9%). Mw: 12.7 kDa.

[0576] Preparation Example P4: Polymer 247

[0577] Polymer 247 contains the following repeating units:

[0578] Following the procedure of Preparation Example P1, the starting materials were A004 (221 mg, 1 eq), B027 (505 mg, 1.05 eq), and cesium fluoride (19.3 mg, 0.1 eq), yielding a yellow solid 247 (367 mg, yield 67.8%). Mw: 10.1 kDa.

[0579] Preparation Example P5: Polymer 262

[0580] Polymer 262 contains the following repeating units:

[0581] Following the procedure of Preparation Example P1, the raw materials were A007 (104 mg, 1 eq), B026 (259 mg, 1.2 eq), and cesium fluoride (9.8 mg, 0.1 eq), yielding white crystalline particles 262 (180 mg, yield 69.2%). Mw: 3.1 kDa.

[0582] Preparation Example P6: Polymer 269

[0583] Polymer 269 contains the following repeating units:

[0584] Following the procedure of Preparation Example P1, the starting materials were A006 (102.1 mg, 1 eq), B003 (260 mg, 1.2 eq), and cesium fluoride (10.3 mg, 0.1 eq), yielding a white oily substance 269 (100 mg, yield 40.5%). Mw: 4.4 kDa.

[0585] Preparation Example P7: Polymer 273

[0586] Polymer 273 contains the following repeating units:

[0587] The starting materials A010 (100 mg, 1 eq), B003 (134 mg, 1.2 eq), and cesium fluoride (5.2 mg, 0.1 eq) were dissolved in 1 mL of ethyl acetate solution and reacted at 60 °C for 15 hours. After the reaction was completed, the mixture was brought to room temperature, and 2 mL of ethyl acetate was added. The mixture was washed three times with pure water (3 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the ethyl acetate was evaporated to obtain a pale yellow oil, 273 (63 mg, yield 36.1%). Mw: 3.5 kDa.

[0588] Preparation Example P8: Polymer 276

[0589] Polymer 276 contains the following repeating units:

[0590] Following the procedure of Preparation Example P1, the starting materials were A004 (106 mg, 1 eq), B003 (222 mg, 1.2 eq), and cesium fluoride (9.4 mg, 0.1 eq), yielding a white oily substance 276 (35 mg, yield 15.2%). Mw: 6.3 kDa.

[0591] Preparation Example P9: Polymer 281

[0592] Polymer 281 contains the following repeating units:

[0593] Following the procedure in Preparation Example P7, the starting materials were A008 (100 mg, 1 eq), B004 (233 mg, 1.2 eq), and cesium fluoride (8 mg, 0.1 eq), yielding a colorless oily product 281 (75 mg, yield 30.6%). Mw: 4.4 kDa.

[0594] Preparation Example P10: Polymer 282

[0595] Polymer 282 contains the following repeating units:

[0596] Following the procedure in Preparation Example P7, the starting materials were A009 (98 mg, 1 eq), B004 (164 mg, 1.2 eq), and cesium fluoride (5.6 mg, 0.1 eq), yielding a colorless oily product 282 (30 mg, yield 14.9%). Mw: 14.1 kDa.

[0597] Preparation Example P11: Polymer 283

[0598] Polymer 283 contains the following repeating units:

[0599] Following the procedure in Preparation Example P7, the starting materials were A010 (100 mg, 1 eq), B004 (155 mg, 1.2 eq), and cesium fluoride (5.3 mg, 0.1 eq), yielding a pale yellow oily product 283 (20 mg, yield 10.2%). Mw: 14.3 kDa.

[0600] Preparation Example P12: Polymer 293

[0601] Polymer 293 contains the following repeating units:

[0602] Following the procedure in Preparation Example P7, the starting materials were A012 (101 mg, 1 eq), B005 (239 mg, 1.2 eq), and cesium fluoride (9.4 mg, 0.1 eq), yielding a pale yellow oily substance 293 (100 mg, yield 41.9%). Mw: 46.1 kDa.

[0603] Preparation Example P13: Polymer 298

[0604] Polymer 298 contains the following repeating units:

[0605] Following the procedure of Preparation Example P1, the starting materials were A032 (100.4 mg, 1 eq), B007 (207 mg, 1.2 eq), and cesium fluoride (7.4 mg, 0.1 eq), yielding a white crystalline solid 298 (160 mg, yield 64.8%). Mw: 5.0 kDa.

[0606] Preparation Example P14: Polymer 299

[0607] Polymer 299 contains the following repeating units:

[0608] Following the procedure in Preparation Example P7, the starting materials were A021 (101 mg, 1 eq), B007 (230 mg, 1.2 eq), and cesium fluoride (8.2 mg, 0.1 eq), yielding a pale yellow oily substance 299 (100 mg, yield 41.4%). Mw: 1.2 kDa.

[0609] Preparation Example P15: Polymer 300

[0610] Polymer 300 contains the following repeating units:

[0611] Following the procedure in Preparation Example P7, the starting materials were A022 (253 mg, 1 eq), B007 (510 mg, 1.2 eq), and cesium fluoride (18.3 mg, 0.1 eq), yielding a pale yellow oily 300 (210 mg, yield 37.2%). Mw: 23.6 kDa.

[0612] Preparation Example P16: Polymer 301

[0613] Polymer 301 contains the following repeating units:

[0614] Following the procedure of Preparation Example P1, the starting materials were A023 (98.5 mg, 1 eq), B007 (260 mg, 1.2 eq), and cesium fluoride (9.3 mg, 0.1 eq), yielding a white crystalline solid 301 (120 mg, yield 46.7%). Mw: 8.5 kDa.

[0615] Preparation Example P17: Polymer 303

[0616] Polymer 303 contains the following repeating units:

[0617] Following the procedure of Preparation Example P1, the starting materials were A032 (100 mg, 1 eq), B005 (192 mg, 1.2 eq), and cesium fluoride (7.4 mg, 0.1 eq), yielding a white, fluffy solid 303 (150 mg, yield 69.5%). Mw: 5.7 kDa.

[0618] Preparation Example P18: Polymer 307

[0619] Polymer 307 contains the following repeating units:

[0620] Following the procedure of Preparation Example P1, the starting materials were A013 (99 mg, 1 eq), B004 (150 mg, 1.2 eq), and cesium fluoride (8.6 mg, 0.1 eq), yielding a white crystalline solid 307 (120 mg, yield 47.0%). Mw: 3.2 kDa.

[0621] Preparation Example P19: Polymer 315

[0622] Polymer 315 contains the following repeating units:

[0623] Following the procedure of Preparation Example P1, the starting materials were A022 (123 mg, 1 eq), B004 (268 mg, 1.2 eq), and cesium fluoride (7.2 mg, 0.1 eq), yielding a yellow oily product 315 (120 mg, yield 41.3%). Mw: 7.5 kDa.

[0624] Following the procedure of Preparation Example P1, the starting materials were A031 (100 mg, 1 eq), B004 (326 mg, 1.2 eq), and cesium fluoride (11.4 mg, 0.1 eq), yielding a white crystalline solid 328 (36 mg, yield 11.8%). Mw: 3.3 kDa.

[0625] Preparation Example P22: Polymer 329

[0626] Polymer 329 contains the following repeating units:

[0627] Following the procedure of Preparation Example P1, the starting materials were A032 (102 mg, 1 eq), B004 (255 mg, 1.2 eq), and cesium fluoride (8.6 mg, 0.1 eq), yielding a white crystalline solid 329 (120 mg, yield 45.9%). Mw: 3.5 kDa.

[0628] Preparation Example P23: Polymer 376

[0629] Polymer 376 contains the following repeating units:

[0630] Following the procedure of Preparation Example P1, the starting materials were A046 (103 mg, 1 eq), B004 (222 mg, 1.2 eq), and cesium fluoride (7.7 mg, 0.1 eq), yielding a white crystalline solid 376 (145 mg, yield 60.0%). Mw: 13.6 kDa.

[0631] Preparation Example P24: Polymer 435

[0632] Polymer 435 contains the following repeating units:

[0633] Preparation Example P20: Polymer 326

[0634] Polymer 326 contains the following repeating units:

[0635] Following the procedure in Preparation Example P7, the raw materials were A029 (97 mg, 1 eq), B005 (170 mg, 1.2 eq), and cesium fluoride (6.7 mg, 0.1 eq), yielding a pale yellow oily substance 326 (83 mg, 42.6%). Mw: 4.1 kDa.

[0636] Preparation Example P21: Polymer 328

[0637] Polymer 328 contains the following repeating units:

[0638] Following the procedure of Preparation Example P1, the starting materials were A017 (101 mg, 1 eq), B004 (210 mg, 1.2 eq), and cesium fluoride (7.2 mg, 0.1 eq), yielding a white crystalline solid 435 (130 mg, yield 55.5%). Mw: 9.3 kDa.

[0639] Preparation Example P25: Polymer 474

[0640] Polymer 474 contains the following repeating units:

[0641] Following the procedure of Preparation Example P1, the raw materials were A006 (100 mg, 1 eq), B021 (305 mg, 1.2 eq), and cesium fluoride (10.0 mg, 0.1 eq), which were lyophilized to obtain a white crystalline solid 474 (40 mg, yield 13.6%). Mw: 1.9 kDa.

[0642] Preparation Example P26: Polymer 475

[0643] Polymer 475 contains the following repeating units:

[0644] Following the procedure in Preparation Example P8, the starting materials were A007 (102 mg, 1 eq), B021 (280 mg, 1.1 eq), and cesium fluoride (9.4 mg, 0.1 eq), yielding a pale yellow oily substance 475 (90 mg, yield 32.1%). Mw: 2.1 kDa.

[0645] Preparation Example P27: Polymer 521

[0646] Polymer 521 contains the following repeating units:

[0647] Raw materials A014 (100 mg, 1 eq), B026 (170 mg, 1.05 eq), and cesium fluoride (7.0 mg, 0.1 eq) were dissolved in 1 mL of dichloromethane solution and reacted at 60 °C for 15 hours. After the reaction was completed, the solution was brought to room temperature. 1 mL of the reaction solution was precipitated with 25 mL of diethyl ether, centrifuged at 4000 x g for 5 minutes, and the supernatant was removed. The solution was redissolved in 1 mL of dichloromethane, precipitated with 25 mL of diethyl ether, centrifuged at 4000 x g for 5 minutes, and the supernatant was removed. The organic phase was evaporated to dryness to obtain a pale yellow solid 521 (88 mg, yield 43.5%). Mw: 16.2 kDa.

[0648] Preparation Example P28: Polymer 522

[0649] Polymer 522 contains the following repeating units:

[0650] Following the procedure in Preparation Example P27, the starting materials were A015 (98 mg, 1 eq), B026 (155 mg, 1.06 eq), and cesium fluoride (6.6 mg, 0.1 eq), yielding a pale yellow solid 522 (75 mg, yield 39.2%). Mw: 17.4 kDa.

[0651] Preparation Example P29: Polymer 523

[0652] Polymer 523 contains the following repeating units:

[0653] Following the procedure in Preparation Example P27, the starting materials were A016 (99 mg, 1 eq), B026 (155 mg, 1.06 eq), and cesium fluoride (6.6 mg, 0.1 eq), yielding a pale yellow solid 523 (140 mg, yield 72.9%). Mw: 18.0 kDa.

[0654] Preparation Example P30: Polymer 524

[0655] Polymer 524 contains the following repeating units:

[0656] Following the procedure in Preparation Example P27, the starting materials were A017 (101 mg, 1 eq), B026 (170 mg, 1.05 eq), and cesium fluoride (7.1 mg, 0.1 eq), yielding a pale yellow solid 524 (100 mg, yield 49.3%). Mw: 12.9 kDa.

[0657] Preparation Example P31: Polymer 525

[0658] Polymer 525 contains the following repeating units:

[0659] Following the procedure in Preparation Example P27, the starting materials were A018 (99 mg, 1 eq), B026 (150 mg, 1.08 eq), and cesium fluoride (6.2 mg, 0.1 eq), yielding a pale yellow solid 525 (135 mg, yield 71.2%). Mw: 12.6 kDa.

[0660] Preparation Example P32: Polymer 526

[0661] Polymer 526 contains the following repeating units:

[0662] Following the procedure in Preparation Example P29, the starting materials were A019 (101 mg, 1 eq), B026 (190 mg, 1.04 eq), and cesium fluoride (8.0 mg, 0.1 eq), yielding a pale yellow solid 526 (80 mg, yield 37.1%). Mw: 11.6 kDa.

[0663] Preparation Example P33: Polymer 528

[0664] Polymer 528 contains the following repeating units:

[0665] Following the procedure in Preparation Example P27, the starting materials were A021 (97 mg, 1 eq), B026 (185 mg, 1.05 eq), and cesium fluoride (8.1 mg, 0.1 eq), yielding a pale yellow solid 528 (90 mg, yield 43.2%). Mw: 14.5 kDa.

[0666] Preparation Example P34: Polymer 547

[0667] Polymer 547 contains the following repeating units:

[0668] Following the procedure in Preparation Example P27, the starting materials were A023 (103 mg, 1 eq), B021 (248 mg, 1.05 eq), and cesium fluoride (9.3 mg, 0.1 eq), yielding a pale yellow solid 547 (150 mg, yield 57.5%). Mw: 17.3 kDa.

[0669] Preparation Example P35: Polymer 550

[0670] Polymer 550 contains the following repeating units:

[0671] Following the procedure in Preparation Example P27, the starting materials were A003 (99 mg, 1 eq), B005 (287 mg, 1.07 eq), and cesium fluoride (12.8 mg, 0.1 eq), yielding a pale yellow solid 550 (110 mg, yield 41.6%). Mw: 15.2 kDa.

[0672] Preparation Example P36: Polymer 609

[0673] Polymer 609 contains the following repeating units:

[0674] Following the procedure in Preparation Example P27, the starting materials were A004 (106.7 mg, 1 eq), B028 (242.7 mg, 1.05 eq), and cesium fluoride (8.7 mg, 0.1 eq), yielding a pale yellow solid 609 (80 mg, yield 23%). Mw: 17.9 kDa.

[0675] Preparation Example P37: Polymer 612

[0676] Polymer 612 contains the following repeating units:

[0677] Following the procedure in Preparation Example P27, the starting materials were A015 (101 mg, 1 eq), B028 (178 mg, 1.05 eq), and cesium fluoride (6.6 mg, 0.1 eq), yielding a pale yellow solid 612 (120 mg, yield 55.6%). Mw: 9.1 kDa.

[0678] Preparation Example P38: Polymer 614

[0679] Polymer 614 contains the following repeating units:

[0680] Following the procedure in Preparation Example P27, the starting materials were A019 (103 mg, 1 eq), B028 (218 mg, 1.05 eq), and cesium fluoride (8.0 mg, 0.1 eq), yielding a pale yellow solid 614 (114 mg, yield 46.9%). Mw: 10.6 kDa.

[0681] Preparation Example P39: Polymer 618

[0682] Polymer 618 contains the following repeating units:

[0683] Following the procedure in Preparation Example P27, the starting materials were A003 (104 mg, 1 eq), B029 (376 mg, 1.05 eq), and cesium fluoride (12.8 mg, 0.1 eq), yielding a pale yellow solid 618 (60 mg, yield 16.9%). Mw: 11 kDa.

[0684] Preparation Example P40: Polymer 619

[0685] Polymer 619 contains the following repeating units:

[0686] Following the procedure in Preparation Example P27, the starting materials were A004 (107 mg, 1 eq), B029 (265 mg, 1.05 eq), and cesium fluoride (8.7 mg, 0.1 eq), yielding a pale yellow solid 619 (86 mg, yield 30.2%). Mw: 8.9 kDa.

[0687] Preparation Example P41: Polymer 723

[0688] Polymer 723 contains the following repeating units:

[0689] Following the procedure of Preparation Example P1, the starting materials were A002 (177 mg, 1 eq), B026 (301 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq). The reaction time was 2 h, yielding a pale yellow solid 723 (160 mg, yield 44.6%). Mw: 6.6 kDa.

[0690] Preparation Example P42: Polymer 726

[0691] Polymer 726 contains the following repeating units:

[0692] Following the procedure of Preparation Example P1, the starting materials were Al119 (207 mg, 1 eq), B026 (301 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 726 (205 mg, yield 52.8%). Mw: 4.4 kDa.

[0693] Preparation Example P43: Polymer 727

[0694] Polymer 727 contains the following repeating units:

[0695] Following the procedure of Preparation Example P1, the starting materials were Al2O (251 mg, 1 eq), B026 (301 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq). The reaction time was 2 h, yielding a pale yellow solid 727727 (207 mg, yield 47.9%). Mw: 6.2 kDa.

[0696] Preparation Example P44: Polymer 728

[0697] Polymer 728 contains the following repeating units:

[0698] Following the procedure of Preparation Example P1, the starting materials were A121 (227 mg, 1 eq), B026 (301 mg, 1 eq), and cesium fluoride (12.8 mg, 0.1 eq), yielding a pale yellow solid 728 (215 mg, yield 52.6%). Mw: 8.2 kDa.

[0699] Preparation Example P45: Polymer 732

[0700] Polymer 732 contains the following repeating units:

[0701] Following the procedure in Preparation Example P27, the starting materials were A086 (153.9 mg, 1 eq), B026 (302 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), yielding a white solid 732 (193 mg, yield 57.4%). Mw: 18.4 kDa.

[0702] Preparation Example P46: Polymer 733

[0703] Polymer 733 contains the following repeating units:

[0704] Following the procedure in Preparation Example P27, the starting materials were A087 (166.9 mg, 1 eq), B026 (302.2 mg, 1 eq), and cesium fluoride (12.8 mg, 0.1 eq), yielding a pale yellow solid 733 (173 mg, yield 49.6%). Mw: 16.6 kDa.

[0705] Preparation Example P47: Polymer 734

[0706] Polymer 734 contains the following repeating units:

[0707] Following the procedure in Preparation Example P27, the starting materials were A088 (179.3 mg, 1 eq), B026 (301.4 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), yielding a pale yellow solid 734 (211 mg, yield 58.5%). Mw: 61.5 kDa.

[0708] Preparation Example P48: Polymer 735

[0709] Polymer 735 contains the following repeating units:

[0710] Following the procedure in Preparation Example P27, the starting materials were A089 (142.5 mg, 1 eq), B026 (303.6 mg, 1 eq), and cesium fluoride (12.6 mg, 0.1 eq), yielding a pale yellow solid 735 (144 mg, yield 44.2%). Mw: 22.4 kDa.

[0711] Preparation Example P49: Polymer 741

[0712] Polymer 741 contains the following repeating units:

[0713] Following the procedure in Preparation Example P27, the starting materials were A095 (141.5 mg, 1 eq), B026 (300.8 mg, 1 eq), and cesium fluoride (12.9 mg, 0.1 eq), yielding a brownish-yellow solid 741 (182 mg, yield 56.3%). Mw: 30.7 kDa.

[0714] Preparation Example P50: Polymer 742

[0715] Polymer 742 contains the following repeating units:

[0716] Following the procedure in Preparation Example P27, the starting materials were A096 (154.9 mg, 1 eq), B026 (300.9 mg, 1 eq), and cesium fluoride (13.3 mg, 0.1 eq), yielding a pale yellow solid 742 (156 mg, yield 46.5%). Mw: 7.2 kDa.

[0717] Preparation Example P51: Polymer 744

[0718] Polymer 744 contains the following repeating units:

[0719] Following the procedure in Preparation Example P27, the starting materials were A098 (178.4 mg, 1 eq), B026 (300.8 mg, 1 eq), and cesium fluoride (12.7 mg, 0.1 eq), yielding a pale yellow solid 744 (152 mg, yield 42.4%). Mw: 4.5 kDa.

[0720] Preparation Example P52: Polymer 745

[0721] Polymer 745 contains the following repeating units:

[0722] Following the procedure in Preparation Example P27, the starting materials were A099 (191.4 mg, 1 eq), B026 (300.6 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), yielding a pale yellow solid 745 (151 mg, yield 40.6%). Mw: 9.2 kDa.

[0723] Preparation Example P53: Polymer 746

[0724] Polymer 746 contains the following repeating units:

[0725] Following the procedure in Preparation Example P27, the starting materials were A100 (203.1 mg, 1 eq), B026 (301.5 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), yielding a pale yellow solid 746 (242 mg, yield 62.8%). Mw: 10.0 kDa.

[0726] Preparation Example P54: Polymer 750

[0727] Polymer 750 contains the following repeating units:

[0728] Following the procedure in Preparation Example P27, the starting materials were A105 (202.8 mg, 1 eq), B026 (302.3 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), yielding a pale yellow solid 750 (245 mg, yield 63.5%). Mw: 9.1 kDa.

[0729] Preparation Example P55: Polymer 751

[0730] Polymer 751 contains the following repeating units:

[0731] Following the procedure in Preparation Example P27, the starting materials were A106 (214.5 mg, 1 eq), B026 (301.8 mg, 1 eq), and cesium fluoride (12.8 mg, 0.1 eq), yielding a pale yellow solid 751 (233 mg, yield 58.7%). Mw: 9.3 kDa.

[0732] Preparation Example P56: Polymer 752

[0733] Polymer 752 contains the following repeating units:

[0734] Following the procedure in Preparation Example P27, the starting materials were A107 (167.3 mg, 1 eq), B026 (302.6 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), yielding a pale yellow solid 752 (196 mg, yield 55.9%). Mw: 6.3 kDa.

[0735] Preparation Example P57: Polymer 756

[0736] Polymer 756 contains the following repeating units:

[0737] Following the procedure in Preparation Example P27, the starting materials were A116 (202.3 mg, 1 eq), B026 (302.9 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), yielding a pale yellow solid 756 (188 mg, yield 48.8%). Mw: 7.8 kDa.

[0738] Preparation Example P58: Polymer 757

[0739] Polymer 757 contains the following repeating units:

[0740] Following the procedure in Preparation Example P27, the starting materials were A117 (216.0 mg, 1 eq), B026 (300.8 mg, 1 eq), and cesium fluoride (12.9 mg, 0.1 eq), yielding a pale yellow solid 757 (159 mg, yield 40.0%). Mw: 9.6 kDa.

[0741] Preparation Example P59: Polymer 758

[0742] Polymer 758 contains the following repeating units:

[0743] Following the procedure in Preparation Example P27, the starting materials were A118 (225.0 mg, 1 eq), B026 (300.2 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), yielding a pale yellow solid 758 (195 mg, yield 48.1%). Mw: 10.4 kDa.

[0744] Preparation Example P60: Polymer 759

[0745] Polymer 759 contains the following repeating units:

[0746] Following the procedure in Preparation Example P27, the starting materials were A084 (118.6 mg, 1 eq), B026 (301.7 mg, 1 eq), and cesium fluoride (13.3 mg, 0.1 eq), yielding a pale yellow solid 759 (154 mg, yield 51.2%). Mw: 12.1 kDa.

[0747] Preparation Example P61: Polymer 761

[0748] Polymer 761 contains the following repeating units:

[0749] Following the procedure of Preparation Example P1, the starting materials were A073 (190.2 mg, 1 eq) and B026 (300.7 mg, 1 eq), and the reaction time was 5 h, yielding a pale yellow solid 761 (164 mg, yield 44.2%). Mw: 10.3 kDa.

[0750] Preparation Example P62: Polymer 763

[0751] Polymer 763 contains the following repeating units:

[0752] Following the procedure of Preparation Example P1, the starting materials were Al22 (238.8 mg, 1 eq), Bo26 (302.3 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 763 (197 mg, yield 46.7%). Mw: 4.5 kDa.

[0753] Preparation Example P63: Polymer 764

[0754] Polymer 764 contains the following repeating units:

[0755] Following the procedure of Preparation Example P1, the starting materials were Al23 (264.6 mg, 1 eq), Bo26 (301.7 mg, 1 eq), and cesium fluoride (13.2 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 764 (256 mg, yield 57.4%). Mw: 3 kDa.

[0756] Preparation Example P64: Polymer 765

[0757] Polymer 765 contains the following repeating units:

[0758] Following the procedure of Preparation Example P1, the starting materials were Al24 (200.0 mg, 1 eq), Bo26 (210.3 mg, 1 eq), and cesium fluoride (13.1 mg, 0.14 eq), and the reaction time was 2 h, yielding a pale yellow solid 765 (102 mg, yield 31.2%). Mw: 2 kDa.

[0759] Preparation Example P65: Polymer 766

[0760] Polymer 766 contains the following repeating units:

[0761] Following the procedure of Preparation Example P1, the starting materials were A074 (142.4 mg, 1 eq), B026 (302.3 mg, 1 eq), and cesium fluoride (12.7 mg, 0.1 eq). The reaction time was 2 h, yielding a pale yellow solid 766 (135 mg, yield 41.5%). Mw: 11.9 kDa.

[0762] Preparation Example P66: Polymer 767

[0763] Polymer 767 contains the following repeating units:

[0764] Following the procedure of Preparation Example P1, the starting materials were A070 (167.0 mg, 1 eq) and B026 (301.8 mg, 1 eq), and the reaction time was 5 h, yielding a white solid 767 (155 mg, yield 44.4%). Mw: 10.4 kDa.

[0765] Preparation Example P67: Polymer 768

[0766] Polymer 768 contains the following repeating units:

[0767] Following the procedure of Preparation Example P1, the starting materials were A071 (155.9 mg, 1 eq), B026 (302.8 mg, 1 eq), and cesium fluoride (13.3 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 768 (161 mg, yield 47.6%). Mw: 5.1 kDa.

[0768] Preparation Example P68: Polymer 769

[0769] Polymer 769 contains the following repeating units:

[0770] Following the procedure of Preparation Example P1, the starting materials were A075 (157.7 mg, 1 eq), B026 (302.1 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 769 (133 mg, yield 39.1%). Mw: 7 kDa.

[0771] Preparation Example P69: Polymer 772

[0772] Polymer 772 contains the following repeating units:

[0773] Following the procedure of Preparation Example P1, the starting materials were A078 (154.8 mg, 1 eq), B026 (303.3 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 772 (124 mg, yield 36.6%). Mw: 6 kDa.

[0774] Preparation Example P70: Polymer 774

[0775] Polymer 774 contains the following repeating units:

[0776] Following the procedure of Preparation Example P1, the starting materials were A082 (165.0 mg, 1 eq), B026 (303.8 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq). The reaction time was 2 h, yielding a white solid 774 (142 mg, yield 40.7%). Mw: 14.8 kDa.

[0777] Preparation Example P71: Polymer 779

[0778] Polymer 779 contains the following repeating units:

[0779] Following the procedure of Preparation Example P1, the starting materials were A128 (193 mg, 1 eq), B026 (300.0 mg, 1 eq), and cesium fluoride (12.9 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 779 (174 mg, yield 46.5%). Mw: 34.7 kDa.

[0780] Preparation Example P72: Polymer 780

[0781] Polymer 780 contains the following repeating units:

[0782] Following the procedure in Preparation Example P27, the starting materials were A129 (203.0 mg, 1 eq), B026 (302.0 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq). The reaction time was 2 h, yielding a pale yellow solid 780 (215 mg, yield 55.9%). Mw: 11.4 kDa.

[0783] Preparation Example P73: Polymer 781

[0784] Polymer 781 contains the following repeating units:

[0785] Following the procedure in Preparation Example P27, the starting materials were A130 (216.0 mg, 1 eq), B026 (301.0 mg, 1 eq), and cesium fluoride (12.8 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 781 (252 mg, yield 63.3%). Mw: 10.1 kDa.

[0786] Preparation Example P74: Polymer 782

[0787] Polymer 782 contains the following repeating units:

[0788] Following the procedure described in Example P27, the starting materials were A131 (180.0 mg, 1 eq), B026 (302.0 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq). The reaction time was 2 h, yielding a pale yellow solid 782 (198 mg, yield 54.7%). Mw: 5.1 kDa.

[0789] Preparation Example P75: Polymer 783

[0790] Polymer 783 contains the following repeating units:

[0791] Following the procedure in Preparation Example P27, the starting materials were A132 (190.3 mg, 1 eq), B026 (301.0 mg, 1 eq), and cesium fluoride (12.7 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 783 (176 mg, yield 47.3%). Mw: 4.9 kDa.

[0792] Preparation Example P76: Polymer 784

[0793] Polymer 784 contains the following repeating units:

[0794] Following the procedure in Preparation Example P27, the starting materials were A133 (205.0 mg, 1 eq), B026 (301.0 mg, 1 eq), and cesium fluoride (13.3 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 784 (226 mg, yield 58.6%). Mw: 13.6 kDa.

[0795] Preparation Example P77: Polymer 790

[0796] Polymer 790 contains the following repeating units:

[0797] Following the procedure of Preparation Example P1, the starting materials were A004 (8.85 g, 1 eq) and B026 (20.00 g, 1,15 eq), and the reaction time was 4 h, yielding a pale yellow solid 790 (15.5 g, yield 55.4%). Mw: 9.8 kDa.

[0798] Preparation Example P78: Polymer 791

[0799] Polymer 791 contains the following repeating units:

[0800] Following the procedure of Preparation Example P1, the starting materials were A003 (27.5 g, 1 eq), B026 (83.00 g, 1.05 eq), and cesium fluoride (7.01 g, 0.2 eq). The reaction time was 5 h, yielding a pale yellow solid 791 (60.7 g, yield 54.2%). Mw: 12.5 kDa.

[0801] Preparation Example P79: Polymer E004

[0802] Polymer E004 contains the following repeating units:

[0803] The starting materials M004 (172 mg, 1 eq) and M005 (525 mg, 1 eq) were dissolved in 2 mL of tetrahydrofuran solution and reacted at 50 °C for 48 hours. After the reaction was completed, the mixture was allowed to return to room temperature. 2 mL of the reaction solution was precipitated with 50 mL of diethyl ether, centrifuged at 4000 x g for 5 minutes, the supernatant was removed, and the organic phase was evaporated to obtain a pale yellow solid product. This solid was then redissolved in 2 mL of dichloromethane, precipitated with 50 mL of diethyl ether, centrifuged at 4000 x g for 5 minutes, the supernatant was removed, and the organic phase was evaporated to obtain a pale yellow solid E004 (383 mg, yield 55%). Mw: 6.2 kDa.

[0804] Preparation Example P80: Polymer C004

[0805] Polymer C004 contains the following repeating units:

[0806] The raw materials M006 (525 mg, 1 eq) and M007 (151 mg, 1 eq) were dissolved in 1 mL of a mixture of methanol and water (methanol / water = 9 / 1) and reacted at 60 °C for 5 days under light-protected and nitrogen-protected conditions. After 5 days, 16 mg of M007 was added and the reaction was continued with stirring for 2 days. After the reaction was completed, the mixture was brought to room temperature and dissolved in 20 mL of 0.1 M hydrochloric acid (HCl) aqueous solution. The solution was then ultrafiltered overnight with HCl aqueous solution at pH = 4 (MWCO: 1000) (ultrafiltration volume 200 mL), concentrated to 10 mL, filtered through a 0.2 μM filter, and lyophilized to obtain a fluffy white solid C004 (323 mg, yield 47.7%). Mw: 5.5 kDa.

[0807] Preparation Example P81: Polymer R003

[0808] Polymer R003 contains the following repeating units:

[0809] The starting materials benzyl alcohol (0.01 g, 0.088 mmol), M008 (0.35 g, 0.98 mmol), and M009 (1 g, 3.45 mmol) were dissolved in 6 mL of anhydrous toluene solution. The system was evacuated for 10 minutes to remove trace amounts of moisture and replaced with nitrogen gas. 100 mg of dried Novozym435 was added, and the mixture was reacted at 70 °C for 24 hours. After the reaction was complete, the mixture was brought to room temperature, diluted with 2 mL of dichloromethane, filtered to remove the enzyme (Novozym435), and precipitated three times with anhydrous diethyl ether. The resulting solid was dried to obtain a white solid R003 (0.87 g, 65% yield). Mw: 8.7 kDa.

[0810] Preparation Example P82: Polymer 250

[0811] Polymer 250 contains the following repeating units:

[0812] Raw materials A003 (48 mg, 1 eq), B003 (148 mg, 1.2 eq), and cesium fluoride (6.0 mg, 0.1 eq) were dissolved in 2 mL of ethyl acetate solution and reacted at 60 °C for 15 hours. After the reaction was completed, the mixture was brought to room temperature, and 2 mL of the reaction solution was precipitated with 50 mL of diethyl ether. The precipitate was centrifuged at 4000 x g for 5 minutes, the supernatant was removed, and the organic phase was evaporated to obtain a pale yellow solid product. The product was dissolved in 20 mL of acidic aqueous solution (pH = 5) and ultrafiltered (MWCO: 1000). After ultrafiltration by 10 times the volume, the solution was concentrated to 10 mL, filtered through a 0.2 μm filter, and lyophilized to obtain a white solid 250 (105 mg, yield 80.6%). Mw: 9.0 kDa.

[0813] Preparation Example P83: Polymer 251

[0814] Polymer 251 contains the following repeating units:

[0815] Following the procedure of Preparation Example P111, A004 (57 mg, 1 eq), B004 (142 mg, 1.2 eq), and cesium fluoride (5.0 mg, 0.1 eq) were used to obtain white crystalline particles 251 (155 mg, yield 82.4.0%). Mw: 10.5 kDa.

[0816] Preparation Example P84: Polymer 792

[0817] Polymer 792 contains the following repeating units:

[0818] Following the procedure described in Example P27, the starting materials were A026 (463.56 mg, 1 eq), B026 (1.23 g, 1, 2 eq), and cesium fluoride (91.30 mg, 0.2 eq). The reaction time was 4 h, yielding a pale yellow solid 792 (0.48 g, yield 39%). Mw: 3.1 kDa.

[0819] Example 3: Characterization of polymer molecular weight and polymer structure molecular arrangement

[0820] Polymers 244, 245, 246, 247, 262, 269, 276, 298, 301, 303, 307, 315, 328, 329, 376, 435, 474, 475, 521, 522, 523, 525, 526, 528, 547, 550, 609, 612, 614, 618, 619, 723, 726, 727, 728, 732, and 73 were prepared using 0.3M sodium acetate buffer (pH 5). 3. Aqueous solutions (5 mg / mL) of polymers 273, 281, 282, 283, 292, 293, 299, 300, 326, 524, and E004 were prepared using 0.3 M sodium acetate buffer (pH 5). Aqueous solutions (5% DMSO, 5 mg / mL) of polymers 273, 281, 282, 283, 292, 293, 299, 300, 326, 524, and E004 were prepared using DMSO-containing solutions. Samples were filtered through a 0.2 μm filter. Molecular weight was determined using aqueous size exclusion chromatography with differential detector and multi-angle static light scattering (SEC-RI-MALS). The mobile phase was 0.3 M sodium acetate buffer (pH 5). The gel chromatography column was an Ultrahydrogel 1000 (Waters). The injector temperature was 4 °C, the column temperature was set to 30 °C, the flow rate was 0.6 mL / min, and the dn / dc ratio was uniformly set to 0.1659.

[0821] use 13 The molecular arrangement of the polymer was characterized by C10 NMR, and the results are shown in Figures 8-11. Polymer 791... 13 The C10 NMR characterization results showed that only one characteristic peak belonging to the carbonyl carbon of the carbonate was observed at 155 ppm, while the carbonyl signals of other carbon arrangements were extremely low (Figures 8 and 9). This confirms that A003 and B026 in the polymer chain are mainly linked in one mode, namely, a regular structure in which A003 and B026 monomers are arranged alternately. The above NMR characterization results, combined with the single-peak distribution obtained by SEC-MALS testing (Mw / Mn = 1.202), confirm the successful synthesis of a copolymer with alternating A003 and A026 arrangements.

[0822] Similarly, polymer 790 13 The C10 NMR characterization results showed a singlet at 155 ppm for the carbonyl group (Figures 10 and 11). Combined with SEC-MALS test results (M... w / M n=1.289 (unimodal distribution), confirming the successful preparation of copolymers with alternating A004 and A026.

[0823] Example 4: Preparation of polymer-nucleic acid complex

[0824] This embodiment provides the following exemplary method for preparing the polymer-nucleic acid complex.

[0825] Method 1: Prepare an aqueous solution (0.8 mg / mL) of the polymer using 20 mM pH 5 sodium citrate buffer. Prepare an aqueous solution (40 μg / mL) of Luc-mRNA (CDS sequence shown in SEQ ID No.:1, full-length sequence shown in SEQ ID No.:2) or pCMV-luc plasmid DNA (PF0461, PlasmidFactory) using 20 mM pH 5 sodium citrate buffer. Mix the polymer solution and the Luc-mRNA solution or pCMV-luc solution thoroughly at a volume ratio of 1:1 and incubate at room temperature for 10 minutes to obtain the polymer / Luc-mRNA or polymer / pCMV-luc complex. Polymers 244, 245, 246, 247, 250, 251, 262, 269, 276, 298, 301, 303, 307, 315, 328, 329, 334, 373, 376, 379, 381, 382, ​​385, 388, 392, 394, 395, 399, 431, 435, 474, 475, 521, 522, 523, 525, 526, 528, 547, 550, 609, 612, 614, 618, 619, 723, 726, 727, 72 Complexes of 8, 732, 733, 734, 735, 741, 742, 744, 745, 746, 750, 751, 752, 756, 757, 758, 759, 761, 763, 764, 765, 766, 767, 768, 769, 772, 774, 779, 780, 781, 782, 783, 784, 790, 791, linear polyethyleneimine (L-PEI, 919012, Merck), C004, and R003 with Luc-mRNA were prepared using method 1. Complexes of linear polyethyleneimine (L-PEI, 919012, Merck), polymers 245, 246, 247, 522, 523, 525, 526, 528, 547, 550, and 791 with the pCMV-luc plasmid were prepared using method 1.

[0826] Method 2: An aqueous solution of the polymer containing DMSO (1% DMSO, 0.8 mg / mL) was prepared using 20 mM pH 5 sodium citrate buffer. An aqueous solution (40 μg / mL) of Luc-mRNA (CDS sequence shown in SEQ ID No.:1, full-length sequence shown in SEQ ID No.:2) was prepared using 20 mM pH 5 sodium citrate buffer. The polymer solution and Luc-mRNA solution or pCMV-luc solution were mixed at a volume ratio of 1:1 and allowed to stand at room temperature for 10 minutes to obtain polymer-Luc-mRNA or polymer / pCMV-luc plasmid complexes. Complexes of polymers 273, 281, 282, 283, 288, 292, 293, 294, 299, 300, 326, 524, and E004 with Luc-mRNA were prepared using Method 2. Complexes of polymers 273, 283, and 524 with pCMV-luc plasmid were prepared using Method 2.

[0827] Example 5: Performance Characterization of Polymer-Nucleic Acid Complexes

[0828] A. Measurement of average particle size, particle dispersion index (PDI), and zeta surface potential of polymer-nucleic acid complex nanoparticles

[0829] 200 μL of the polymer / Luc-mRNA or polymer / pCMV-luc complex prepared in Example 4 was added to a micro-sample cell (ZEN0040). The nanoparticle size and particle dispersion coefficient of the sample were measured using a nanoparticle size analyzer (Zetasizer Pro, Malvern Panalytical) at 25°C and a laser angle of 173°. 1 mL of the polymer / Luc-mRNA or polymer / pCMV-luc complex prepared in Example 4 was taken using a 1 mL syringe and added to a disposable foldable capillary sample cell (DTS1070). The surface potential (Zeta) of the sample was measured using a Zetasizer Pro.

[0830] B. Measurement of polymer-nucleic acid complex encapsulation efficiency

[0831] Take the polymer / nucleic acid complex prepared in Example 4, 400 ng of Luc-mRNA or pCMV-luc, and premix it with 10× DNA buffer (P022-02-AA, Vazyme) at a volume ratio of 9:1. Take 20 μL of the premix and load it onto a 1% pre-prepared agarose gel (G661012, Thermo Scientific). TMThe gel was run in 1×TEA electrophoresis buffer (abs9260, Absin (Shanghai) Biotechnology Co., Ltd.) at 150V for 15 minutes using a voltage of 150V (1645050, 1704486, Bio-RAD). The gel was then removed and imaged using a UV imager (4600SF, Tanon). Normalization was performed using the gel brightness of unencapsulated Luc-mRNA or pCMV-luc. After normalization, if no bright band appeared in the sample, the encapsulation rate of the complex was recorded as 100%. The results are shown in Table 4 or Table 5.

[0832] Table 4. Characterization results of the polymer-Luc-mRNA complex

[0833] Table 5. Characterization results of the polymer-pCMV-luc plasmid complex

[0834] Example 6: Cell safety and transfection performance of the complex

[0835] The safety and transfection performance of the complex were verified by mRNA transfection using healthy human primary bronchial cells (HBE, ID: BML31M2, passages 5-11), and by plasmid transfection using HEK-293T cells (CBP60439, ATCC, passages 4-7). The culture conditions and medium formulations for primary bronchial HBE cells were set according to the reference (Eur Respir J 2022; 59:2100671). HEK-293T cells were cultured in DMEM medium (GIBCO catalog number: 10566016) containing 10% fetal bovine serum (GIBCO catalog number: A5256701).

[0836] 3×10 4 One HBE cell or 2×10 4 HEK-293T cells were seeded in 96-well cell culture plates (Cat: 167425, Thermo Scientific). TMThe cells were incubated in a 37°C, 5% CO2 incubator for 24 hours. 5 μL of PBS (PB180237, Procell, negative control) or 5 μL of the polymer / Luc-mRNA complex or polymer / pCMV-luc complex prepared in Example 4 (including complexes of polymer L-PEI, polymer E004, polymer C004, and polymer R003 with Luc-mRNA or pCMV-luc as comparative examples) were added directly to the cell culture medium in each well of a 96-well plate and incubated for another 15 hours (Luc-mRNA) or 48 hours (pCMV-luc) in a 37°C, 5% CO2 incubator.

[0837] Cell viability after complex treatment was tested to reflect the safety performance of the complex. Prestoblue solution from cell culture medium (PneumaCult-ALI basal medium, Cat:05002, Stemcell Technologies) and the Prestoblue HS cell viability assay kit (P50201, Invitrogen) was mixed at a 9:1 volume ratio to prepare the Prestoblue mixture. The culture medium was removed from each well of a 96-well cell culture plate, and 100 μL of the Prestoblue mixture was added. The plate was then incubated at 37°C, 5% CO2 for 30 minutes. Subsequently, 80 μL of the mixture was transferred from each well to a 96-well black agar plate (Cat:237107, Thermo Scientific). TM The Varioskan LUX multi-functional microplate reader (VLBL00GD1, Thermo Scientific) was used. TM Fluorescence readings were tested with excitation at 560 nm and emission at 590 nm. Readings from culture wells containing PBS samples were set to 100%. Readings from complex transfection wells were normalized to PBS sample readings. Results are presented as relative cell viability (%Cell Viability), shown in Figures 2 and 3 (after Luc-mRNA transfection) and Figure 4 (after pCMV-luc transfection). This demonstrates the good biocompatibility of the polymer-nucleic acid complex on cells.

[0838] Remove any remaining Prestoblue solution from the 96-well cell culture plate and add 100 μL of 1X cell lysis buffer (E1531, Promega) to each well. Incubate at 4°C for 10 minutes. Transfer 20 μL of lysis buffer to a white 96-well plate (Cat: 236107, Thermo Scientific). TMAdd 50 μL of Luciferase 1000 assay reagent (E4550, Promega) to the medium and incubate at room temperature for 5 minutes. Read the biofluorescence signal (RLU) using a Varioskan LUX multi-functional microplate reader. The results are shown in Figures 5 and 6 (after Luc-mRNA transfection) and Figure 7 (after pCMV-luc transfection). The results show that the polymer-nucleic acid complex effectively transfects cells and successfully expresses the loaded nucleic acid, with a transfection efficiency significantly higher than the negative control and all comparative examples.

[0839] Example 7: In vivo transfection performance of the complex

[0840] Female BALB / c mice (6-8 weeks old, weighing 15-20g) were purchased from Pengyue Biotechnology Co., Ltd. (Jinan, China). All mice were housed in the animal facility of the Institute of Materia Medica, Shandong Academy of Medical Sciences, under SPF conditions: 12h light / 12h dark cycle, temperature 18-23℃, and humidity 40-60%. All animal experiments were conducted in accordance with the "Guidelines for the Husbandry, Management and Use of Laboratory Animals" and approved by the Ethics Committee of the Institute of Materia Medica, Shandong Academy of Medical Sciences (Jinan, China).

[0841] 1. Study on mRNA expression after intramuscular injection of polycarbonate-mRNA complex

[0842] Following the method described in Example 4, different polycarbonate-Luc mRNA complexes (with a final Luc mRNA concentration of 1000 μg / mL) were prepared and diluted to a final mRNA concentration of 200 μg / mL for later use. BALB / c mice were randomly divided into three groups of three mice each. Each mouse was administered 50 μL of the polycarbonate-Luc mRNA complex solution (10 μg Luc mRNA) via intramuscular injection. Twenty-four hours after administration, 100 μL of luciferase substrate was injected intraperitoneally into the mice, followed by gas anesthesia with isoflurane six minutes later. Finally, in vivo imaging was performed using the IVIS Lumina III small animal in vivo imaging system, and the luminescence intensity of the bioluminescent sites was quantitatively analyzed. The results are shown in Figure 12. After intramuscular injection of polymers 790 and 791 with the Luc-mRNA complex, strong bioluminescent signals were observed at the injection site, indicating successful expression of the mRNA-encoded luciferase, and its expression level was significantly higher than that of the L-PEI / Luc-mRNA complex.

[0843] 2. Time-dependent study of mRNA expression after intramuscular injection of polycarbonate-mRNA complex

[0844] Following the method described in Example 4, a polycarbonate-mRNA complex (with a final Luc mRNA concentration of 1000 μg / mL) was prepared and diluted to a final mRNA concentration of 200 μg / mL. BALB / c mice were randomly divided into two groups of three each. Each mouse was injected intramuscularly with 50 μL of the polycarbonate-mRNA complex solution (10 μg Luc mRNA). At 6 h, 24 h, 48 h, 120 h, 168 h, and 336 h post-administration, mice were intraperitoneally injected with 100 μL of luciferase substrate, followed by gas anesthesia with isoflurane 6 min later. Finally, in vivo imaging was performed using the IVIS Lumina III small animal in vivo imaging system, and the luminescence intensity of the bioluminescent sites was quantitatively analyzed.

[0845] The results are shown in Figure 13. Polymers 790 and 791 and the Luc-mRNA complex continuously and stably expressed luciferase protein at the injection site, with significant bioluminescent signals, and the expression lasted for up to 336 hours.

[0846] 3. Evaluation of the transfection efficacy of polycarbonate-Luc-mRNA complex administered intratumorally.

[0847] 1×10 6 One CT26 cell line with the luciferase gene knocked out was suspended in 100 μL of PBS and subcutaneously injected into the groin region of mice. After injection, tumor size was measured periodically using calipers, and tumor volume V (in mm) was calculated using the formula V = (π / 6)LS². 3 (where L is the maximum surface diameter of the tumor and S is the minimum surface diameter). Mice were included in the study when the tumor volume reached approximately 50-100 mm³ (usually 7-14 days after tumor cell injection). Appropriate endpoints were set according to experimental design and animal ethics requirements; the experiment was terminated when the tumor reached a certain volume or significant discomfort occurred. A polycarbonate-mRNA complex (Luc mRNA concentration of 1000 μg / mL) was prepared according to the method in Example 4 and diluted to an mRNA concentration of 500 μg / mL. The tumor volume was 50-100 mm³. 3BALB / c mice with tumor grafts were randomly divided into 5 groups of 3 mice each. Each mouse was injected intratumorally with 20 μL of polycarbonate-mRNA complex solution (10 μg Luc-mRNA). In vivo imaging was performed on the mice at 4 h and 24 h post-administration, and the bioluminescence intensity of the tumor sites was quantitatively analyzed. Before imaging, 100 μL of luciferase substrate was injected intraperitoneally into the mice, and they were anesthetized with isoflurane 6 minutes later. In vivo imaging was performed using the IVIS Lumina III small animal in vivo imaging system, and the fluorescence intensity of the bioluminescent sites was quantitatively analyzed. The group transfected with the LNPs-mRNA complex (prepared with aminocationic lipid SM-102 according to the operating procedure) served as a positive control.

[0848] All data were statistically analyzed using GraphPad Prism 10.0 software (GraphPad Software, La Jolla, USA). Data are expressed as mean ± standard deviation (Mean ± SD). Differences between groups were compared using one-way ANOVA. Independent samples t-tests were used to compare two groups. The significance level for all statistical analyses was set at p < 0.05.

[0849] The results, shown in Figure 14, indicate that both polymer 790 and 791 Luc-mRNA complexes showed luciferase protein expression only at the tumor site, demonstrating good tumor tissue specificity. The protein expression level of the polymer 790 complex within the tumor was significantly higher than that of the 791 complex (Figure 14B). Notably, although the protein expression level of the LNPs-mRNA complex within the tumor was comparable to that of the 790 complex, the LNPs-mRNA complex exhibited significant non-specific ectopic expression in the liver, with its 24-hour expression level in the liver significantly higher than that in the tumor site (Figure 14C). This result suggests that the polymer 790 carrier possesses superior tumor-targeting specificity compared to the LNPs system.

[0850] Unless otherwise expressly stated, all dimensions and their specific values ​​mentioned herein should not be construed as strictly limited to the precise values ​​stated. Rather, each value should be understood to include the value itself and its neighboring functionally equivalent range of variation. For example, a dimension disclosed as “10 nm” should be understood to mean “approximately 10 nm”.

[0851] Unless expressly excluded or otherwise limited, all references cited herein, including any cross-referenced or related patents or patent applications, and any patents or patent applications claiming priority or beneficial effects thereof, are incorporated herein by reference in their entirety. Reference to any document does not constitute an admission that it is prior art, whether alone or in combination with any disclosure or claimed protection of this invention; nor should it be construed as an admission that the document, on its own or in combination with any one or more references, proposes, or discloses any aspect of this invention. Furthermore, where the meaning or definition of a term in this application differs from the meaning or definition of the same term in any cited document, the meaning or definition given to it in this application shall prevail.

[0852] Although the invention has been described and illustrated through specific embodiments, those skilled in the art will understand that various modifications, substitutions, and alterations can be made without departing from the essential spirit and scope of the invention. Therefore, this specification is intended to cover all modifications and equivalent embodiments falling within the scope defined by the appended claims.

Claims

A polycarbonate or salt thereof, the polycarbonate having repeat units of Formula I: wherein, X and Y each independently comprise at least one nitrogen-containing group and / or at least one redox-sensitive group, with the proviso that at least one of X and Y comprises at least one protonatable nitrogen-containing group, and X is different from Y; L is a bond, C 1-16 alkylene or a carbonate bond; R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl; a1 and a4 are each independently an integer from 1 to 16; a2 and a3 are each independently an integer from 0 to 16; and b1 to b4 are each independently an integer from 0 to 2. The polycarbonate or salt thereof according to claim 1, wherein, the at least one nitrogen-containing group is located in the main chain of the polycarbonate; specifically, at least one nitrogen atom in the at least one nitrogen-containing group is located in the main chain of the polycarbonate; more specifically, the at least one protonatable nitrogen-containing group is located in the main chain of the polycarbonate; more specifically, at least one nitrogen atom in the at least one protonatable nitrogen-containing group is located in the main chain of the polycarbonate. The polycarbonate or salt thereof according to claim 1, wherein, the at least one protonatable nitrogen-containing group has a group selected from the group consisting of an amino group, a nitrogen-containing cyclic group, and a combination thereof; specifically, the amino group is selected from the group consisting of a secondary amino group, a tertiary amino group, a quaternary amino group, and a combination thereof; specifically, the nitrogen-containing cyclic group is selected from the group consisting of a 3- to 10-membered saturated or unsaturated aliphatic or aromatic heterocyclic group having at least one ring nitrogen atom, a 5- to 20-membered heterocyclic group fused with each other or with an aliphatic or aromatic carbocyclic group from the 3- to 10-membered saturated or unsaturated aliphatic or aromatic heterocyclic group having at least one ring nitrogen atom, and a combination thereof. The polycarbonate or salt thereof according to claim 1, wherein, at least one of the at least one redox-sensitive group is located in the main chain of the polycarbonate; specifically, the at least one redox-sensitive group comprises one or more of a single sulfur bond, a single selenium bond, a double sulfur bond, a double selenium bond, a triple sulfur bond, a triple selenium bond, a quadruple sulfur bond, a ketone thioacetal bond. The polycarbonate or salt thereof according to claim 1, wherein, The polycarbonate has a weight average molecular weight (Mw) of 0.5 to 100 kDa. w ) A method of making polycarbonate comprising using a polymerization reaction of a monomer having formula II with a monomer having formula III: in formula II and formula III, X 1 and X 2 each independently is: a bond or Y 1 and Y 2 each independently is: a bond or X and Y each independently comprise at least one nitrogen-containing group and / or at least one redox-sensitive group; L is a bond, C 1-16 alkylene or a carbonate bond; R 1 Each group is independently selected from the following groups when it appears: hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester group, substituted or unsubstituted C group. 1-12 Alkyl, substituted or unsubstituted C 1-12 Alkoxy, substituted or unsubstituted C 1-12 Alkylamino, substituted or unsubstituted C 1-12 Alkyl ester group, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted 3- to 10-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl; b1 to b4 are each independently an integer from 0 to 2. a1 and a4 are each independently an integer from 1 to 16; and a2 and a3 are each independently an integer from 0 to 16; with the proviso that: (1) formula II and formula III together have at least one X and at least one Y, and the at least one X and the at least one Y do not come from formula II or formula III at the same time; (2) at least one of X and Y comprises at least one protonatable nitrogen-containing group, and X is different from Y; wherein, wavy line represents a bonding position to an adjacent atom. A complex of the polycarbonate or salt thereof according to claim 1 and at least one nucleic acid. The complex according to claim 7, wherein, The at least one nucleic acid is selected from the group consisting of a deoxyribonucleic acid (DNA), a ribonucleic acid (RNA), a peptide nucleic acid (PNA), a locked nucleic acid (LNA), and any combination thereof. The complex according to claim 7, wherein, The complexing mass ratio of the polycarbonate or salt thereof to the at least one nucleic acid ranges from 0.1:1 to 500:

1. A composition comprising the polycarbonate or salt thereof according to claim 1, or the complex according to claim 7; specifically, a pharmaceutical composition. A method of delivering or expressing at least one nucleic acid in a cell or a subject, the method comprising: providing the complex according to claim 7, which is formed by complexing the at least one nucleic acid with the polycarbonate or salt thereof; and and - contacting said cell or said subject with said complex. Use of the polycarbonate or salt thereof according to claim 1 as a nucleic acid carrier for delivering said nucleic acid to a cell or a subject.

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