Functional polycarbonate, and preparation method therefor and use thereof

Abstract

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

Description

Functional polycarbonates and methods of making and using the same TECHNICAL FIELD

[0001] The present application relates to the field of functional polymer materials; in particular, to a novel ultrafast-degradable polycarbonate nanocarrier, a preparation method thereof, a complex of the nanocarrier and a nucleic acid, and a use of the nanocarrier for delivering the nucleic acid. BACKGROUND

[0002] Nucleic acids, such as DNA and RNA, are large biomolecules responsible for the carrying and delivery of genetic information in cells of an organism. In the treatment of numerous diseases, delivering a desired nucleic acid molecule into a target cell is a core step to achieve the therapeutic purpose. However, the poor in-vitro stability and weak ability to cross the cell membrane of nucleic acid molecules make the development and application of nucleic acid delivery carriers a major demand in the field of nucleic acid therapy.

[0003] Viral vectors were initially developed for nucleic acid delivery due to their high cell infection ability. However, the risk of in-vivo replication, adverse immune response, insertion mutation risk, complex preparation process, and low loading capacity of viral vectors have seriously restricted their large-scale production and industrial application.

[0004] As an alternative to viral vectors, various non-viral carrier systems have been studied, including lipid nanoparticles (LNP), polymeric nanoparticles (PNP), etc. LNP, as the most in-depth researched and most widely used delivery carrier in recent years, has made remarkable achievements in mRNA vaccine delivery. However, its stability, storage conditions, and delivery efficiency still need to be optimized. At the same time, the in-vivo adverse reactions of some components of LNP have also raised concerns and disputes about safety. In contrast, the research and application of PNP have developed more slowly. The biocompatibility and biodegradability of PNP vary depending on the type and structure of the polymer, and some polymers (such as PEI) can accumulate in the body, causing long-term side effects. In addition, the effectiveness of nucleic acid delivery by PNP also needs to be improved to ensure that sufficient amounts of nucleic acids can reach the target site and function.

[0005] There is an urgent need in the art for a novel carrier that is easy to produce on a large scale and can safely and effectively deliver various types of nucleic acids. SUMMARY

[0006] After long-term extensive exploration and research, the inventors have found a novel polymeric nanocarrier that overcomes the problems of low delivery efficiency, high biological toxicity, and complex preparation process that are prevalent but long unresolved in existing carriers, providing a safe, effective, and reliable means for nucleic acid delivery, suitable for large-scale production and industrial application.

[0007] In one aspect, provided herein is a polycarbonate or a salt thereof, the polycarbonate having a repeat unit of Formula I:

[0008] wherein,

[0009] A comprises at least one nitrogen-containing group;

[0010] R 1 each occurrence is independently selected from the group consisting of hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, 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, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 aryl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0011] each of a1and a2is independently an integer from 1 to 16;

[0012] each of b1and b2is independently an integer from 0 to 2.

[0013] In another aspect, provided herein is a method of making a polycarbonate, comprising performing a polymerization reaction using at least one monomer having Formula II and at least one monomer having Formula III:

[0014] in Formula II and Formula III,

[0015] A 1 and A 2 each independently comprises one or more groups independently having Formula IV:

[0016] wherein,

[0017] M is selected from the group consisting of a nitrogen-containing group, a redox-sensitive group, an alkylene group, an oxygen-containing group, and combinations thereof;

[0018] each of x1and x2is independently an integer from 0 to 2;

[0019] each of y1and y2is independently an integer from 0 to 16;

[0020] R 1 each occurrence is independently selected from the group consisting of hydrogen, deuterium, tritium, hydroxyl, halogen, carboxyl, nitro, amino, cyano, substituted or unsubstituted ester, substituted or unsubstituted C 1-12 alkyl, substituted or unsubstituted C 1-12alkoxy, substituted or unsubstituted C 1-12 alkylamino, substituted or unsubstituted C 1-12 alkyl ester, substituted or unsubstituted C 3-14 cycloalkyl, substituted or unsubstituted C 6-14 aryl, substituted or unsubstituted 3- to 10-membered heterocycloalkyl, and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0021] wherein A 1 and A 2 has a nitrogen-containing group;

[0022] wherein, for A 1 and A 2 when comprising two or more groups independently having Formula IV, the two or more groups independently having Formula IV are connected to each other via L M wherein L M is a bond, a carbonate bond, or a C 1-16 alkylene group; and

[0023] wherein the wavy line represents a bonding position to an adjacent atom.

[0024] In another aspect, provided herein is a complex of the polycarbonate or salt thereof and at least one nucleic acid.

[0025] In another aspect, provided herein is a method of making a polycarbonate nucleic acid complex, comprising:

[0026] - providing a polycarbonate or salt thereof, and at least one nucleic acid; and

[0027] - mixing the polycarbonate or salt thereof with the at least one nucleic acid.

[0028] In another aspect, provided herein is a composition comprising the polycarbonate or salt thereof, or the complex.

[0029] In another aspect, provided herein is a product comprising:

[0030] - the complex;

[0031] - the composition; or

[0032] - the polycarbonate or salt thereof and at least one nucleic acid.

[0033] In another aspect, provided herein is a method of delivering at least one nucleic acid to a cell or a subject, the method comprising:

[0034] - providing the complex formed by complexing a polycarbonate or salt thereof with the at least one nucleic acid;

[0035] and

[0036] - contacting the cell or the subject with the complex.

[0037] In another aspect, provided herein is a method of expressing at least one nucleic acid in a cell or a subject, the method comprising:

[0038] - providing the complex, the complex being complexed from a polycarbonate or a salt thereof and the at least one nucleic acid;

[0039] and

[0040] - contacting the cell or the subject with the complex.

[0041] In another aspect, provided herein is the use of the polycarbonate or the salt thereof as a nucleic acid carrier for delivering the nucleic acid to a cell or a subject.

[0042] In another aspect, provided herein is the use of the polycarbonate or the salt thereof, complex or composition for delivering at least one nucleic acid to a cell or a subject.

[0043] In another aspect, provided herein is the use of the polycarbonate or the salt thereof, complex or composition for causing a cell or a subject to express at least one nucleic acid.

[0044] In another aspect, provided herein is the use of the polycarbonate or the salt thereof, complex or composition in the manufacture of a product for delivering at least one nucleic acid to a cell or a subject.

[0045] In another aspect, provided herein is the use of the polycarbonate or the salt thereof, complex or composition in the manufacture of a product for causing a cell or a subject to express at least one nucleic acid. BRIEF DESCRIPTION OF DRAWINGS

[0046] The present application is further described by the following drawings in which:

[0047] Figure 1 shows a schematic diagram of a polycarbonate according to herein forming a polycarbonate nucleic acid complex with a nucleic acid.

[0048] Figure 2 shows cell safety test results with an exemplary polycarbonate / Luc-mRNA complex according to herein.

[0049] Figure 3 shows cell safety test results with an exemplary polycarbonate / Luc-mRNA complex according to herein.

[0050] Figure 4 shows cell safety test results with an exemplary polycarbonate / pCMV-luc complex according to herein.

[0051] Figure 5 shows cell transfection results with exemplary polycarbonate / Luc-mRNA complexes according to the present teachings.

[0052] Figure 6 shows cell transfection results with exemplary polycarbonate / Luc-mRNA complexes according to the present teachings.

[0053] Figure 7 shows cell transfection results with exemplary polycarbonate / pCMV-luc complexes according to the present teachings.

[0054] Figure 8 shows the CDS nucleotide sequence of Luc-mRNA used to prepare polycarbonate / Luc-mRNA complexes (SEQ ID No.: 1).

[0055] Figure 9 shows the full-length nucleotide sequence of Luc-mRNA used to prepare polycarbonate / Luc-mRNA complexes (SEQ ID No.: 2). DETAILED DESCRIPTION

[0056] Nucleic acid molecules, as the substance that carries and transmits genetic information of organisms, are one of the most critical biological macromolecules in cells. DNA is responsible for storing genetic code and guiding cell activities, while RNA is involved in various biological processes, including as a messenger of genetic information (mRNA), participating in protein synthesis (tRNA and rRNA), and regulating gene expression (small RNA). In modern medicine, by delivering specific nucleic acid drugs to the target site, antigen proteins can be expressed to stimulate immune responses, or genetic information that causes adverse consequences can be regulated or corrected to prevent or treat various diseases or conditions, including inflammatory conditions, cancer, genetic diseases, etc.

[0057] However, despite the huge market potential of nucleic acid drugs, their actual application is hindered by many obstacles. Nucleic acid molecules are extremely susceptible to degradation before reaching the target site, and often cannot effectively cross biological barriers (such as cell membranes) to enter the interior of cells, because their large molecular weight and negative charge limit their ability to pass through hydrophobic cell membranes.

[0058] As an example of messenger RNA (mRNA), as a transient, encodable genetic information carrier transcribed from a DNA strand and carrying the encoding information for protein synthesis, mRNA can guide the translation and expression of functional proteins in cells. mRNA-based therapy can produce almost any functional protein by delivering mRNA encoding different proteins into the body for translation and expression, and thus is expected to be used for the treatment of various (intractable) diseases, including infectious diseases, metabolic genetic diseases, cancers, cardiovascular and cerebrovascular diseases, etc. Recently, two mRNA vaccines (mRNA-1273 and BNT162b2) produced by Moderna and Pfizer-BioNTech have been successfully used to prevent COVID-19, highlighting the great potential of mRNA technology in revolutionizing life sciences and medical research. In addition to COVID-19 vaccines, in recent years, there have been a number of mRNA vaccines against other infectious diseases (such as respiratory syncytial virus, seasonal influenza, rabies virus), cancers (such as melanoma), as well as mRNA drugs for protein therapy and gene editing in different clinical trial stages. With the breakthrough development of nucleic acid chemistry and RNA biology technology, such as the modification of traditional linear mRNA untranslated region (UTR) and the emergence of circular mRNA, the most concerned problems of immunogenicity and protein expression effect in mRNA have been largely solved. Therefore, the challenge of clinical translation of the therapeutic potential of mRNA drugs mainly comes from the development of mRNA delivery carriers.

[0059] However, mRNA is a polyanionic compound with a large molecular weight, which is not easy to cross the non-polar cell membrane and tissue barrier. In addition, it is easily inactivated by rapid destruction by nucleases. Therefore, the protection and endosome escape of the delivery carrier are needed to deliver the functional mRNA into the cytoplasm to exert the therapeutic effect. Due to poor degradation performance, the existing mRNA delivery carriers generally have problems of toxicity and delayed nucleic acid release (low transfection efficiency). Specifically, if the delivery carrier is not degradable or the degradation product has high toxicity, safety problems will be caused; if the carrier degrades too slowly, the mRNA cannot be released quickly, which restricts the transfection efficiency.

[0060] Therefore, the development of safe and effective nucleic acid delivery carriers has become the biggest bottleneck restricting the release of the application potential of current nucleic acid drugs. The successful development of such carriers not only can greatly improve the delivery efficiency and thus improve the efficacy of nucleic acids, but also can be widely used and popularized in many fields due to the 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 the efficacy and quality of life of patients.

[0061] After long-term extensive exploration and research, the inventors have innovatively developed a new type of carbonate polymer, which can be used as a polymer nanocarrier for delivering nucleic acids. The polymer nanocarrier is stable in vitro, and can be rapidly and completely self-degraded after entering the target position (such as cells) in experiments or pharmacology, and the degradation products are all small molecules with high safety (such as CO2, thiol, etc.). Due to its unique physicochemical properties, the material can form a complex with a wide range of molecular weights and lengths and different types of nucleic acids, effectively protecting them from enzymatic degradation, enhancing their stability, and promoting the transport across the cell membrane, thereby significantly improving the delivery efficiency of the target nucleic acid molecules. Moreover, the nanomaterial has a simple and environmentally friendly preparation process, and is suitable for large-scale production to meet the needs of extensive experimental and industrial applications, thereby providing a new choice for the field of nucleic acid delivery.

[0062] Definitions

[0063] To facilitate better understanding of the present disclosure, some key terms are defined first. Unless otherwise explicitly stated, each of the terms listed below shall be understood based on the appended definitions. Unless otherwise stated, the meaning of other terms herein is consistent with the general understanding of those skilled in the art.

[0064] Herein, "one" or a combination thereof with various quantifiers includes both singular and plural meanings. Herein, when multiple numerical values, numerical value ranges or combinations thereof are given to explain the same parameter or variable, it is equivalent to specifically disclosing these numerical values, range end values and numerical value ranges formed by any combination thereof. Herein, any numerical value, whether or not with a modifier such as "about", always covers the approximate range that can be understood by those skilled in the art, such as ±20%, ±10%, ±5%, etc.

[0065] Herein, each "embodiment" equally refers to and encompasses the embodiments of the methods and systems herein. One or more technical features in any embodiment can be freely combined with one or more technical features in any one or more other embodiments, and the resulting embodiments also belong to the disclosure herein.

[0066] Unless otherwise specified, all percentages and ratios used herein are by weight. Unless otherwise specified, all temperatures are in degrees Celsius (°C). Room temperature or ambient temperature refers to a temperature of 20°C to 28°C (e.g., 25°C). Unless otherwise specified, all measurements are understood to be at ambient conditions, i.e., at room temperature, about one atmosphere of pressure, and about 50% relative humidity.

[0067] All numerical ranges are inclusive of narrower ranges. The upper and lower limits of the ranges described can be combined with one another to form additional ranges not expressly described.

[0068] As used herein, the term "polycarbonate" refers to a class of polymers in which at least two repeat units are linked by carbonate linkages -OC(O)O-. The terms "polycarbonate", "carbonate polymer" are used interchangeably herein and mean the same thing. It is understood that when "polycarbonate" is referred to herein, it means polycarbonates and salt forms thereof. The salt forms of the polycarbonates are formed by salt formation with anions such as chloride, acetate, citrate, tetrafluoroacetate, and the like.

[0069] As used herein, the term "polymer" refers to a high molecular weight compound formed by the linking of repeat monomer units by covalent bonds. The repeat monomer units can be the same, forming a "homopolymer", or different, forming a "copolymer".

[0070] As used herein, the term "repeat unit" refers to a polymerized unit that occurs at least twice in the polymer backbone. Repeat units are typically derived from monomers. The term "monomer" refers to a small molecule compound that is capable of linking into a long chain molecule through polymerization. During polymerization, multiple chemical bonds of the monomer molecules are broken and reformed, resulting in the repeat linkage of monomer units to form the backbone of the polymer.

[0071] As used herein, the term "linear polymer" refers to a high molecular weight compound having a linear structure for the molecular chain. Linear polymers have relatively small side groups that are not considered branching structures. The term "crosslinked polymer" refers to a polymer having a three-dimensional network structure (e.g., via crosslinking reactions).

[0072] As used herein, the term "random copolymer" refers to a product obtained by random copolymerization, in which two or more monomer units are randomly distributed in the backbone, and no monomer unit forms a separate long segment in the polymer chain. The term "block copolymer" refers to a copolymer in which two or more monomer units each form long sequential segments that are covalently bound to each other.

[0073] As used herein, the term "backbone" refers to the longest linear molecular chain that makes up a polymer. All other molecular chains can be considered side groups or side chains with respect to the backbone. For example, the backbone of a polymer can be composed primarily of carbon atoms linked by covalent bonds, and can be interrupted by nitrogen, oxygen, sulfur, and the like. Generally, end groups present at the ends of the polymer are not included in the backbone. When referring to a "having" or "being located in the backbone" a group or atom, it is meant that the group or atom makes up part of the polymer backbone (and not a side group or end group). It can be considered that a break at the location of a group or atom "located in the backbone" would result in a break in the polymer molecular chain.

[0074] As used herein, the term "amino" refers to the functional group of an amine, and can include primary, secondary, tertiary, quaternary amines.

[0075] 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.

[0076] 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] 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.

[0081] 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).

[0082] 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).

[0083] 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.

[0084] 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).

[0085] 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.

[0086] In this article, the term "halogen" includes fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0087] 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.

[0088] In this article, the term "amide group" refers to a group represented by any of the following chemical formulas: -CONH2, -CONH-, -CON<.

[0089] 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 C 6-20 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.

[0090] In this document, the term "redox-sensitive group" refers to a group whose molecular structure is responsive to one or both of an oxidizing or reducing environment; 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.

[0091] In this document, the term "complex" refers to the product obtained by combining polycarbonate 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 in this document.

[0092] 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).

[0093] 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.

[0094] In this document, "pharmaceutically acceptable" means a substance suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, within the bounds of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio. The term "pharmaceutically acceptable delivery vehicle" includes pharmaceutically acceptable substances, compositions, or carriers suitable for administration to mammals of the polycarbonate or polycarbonate complexes described herein.

[0095] 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.

[0096] polycarbonate

[0097] On the one hand, this article provides a polycarbonate or a salt thereof having nitrogen-containing repeating units.

[0098] Specifically, the polycarbonate or a salt thereof has repeating units of Formula I:

[0099] in,

[0100] A contains at least one nitrogen-containing group;

[0101] 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 7-membered heterocyclic alkyl and substituted or unsubstituted 5- to 14-membered heteroaryl;

[0102] a1 and a2 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.

[0103] b1 and b2 are each an integer from 0 to 2, for example, 0, 1 or 2.

[0104] 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 atom of the at least one nitrogen-containing group is 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 may contain no more than 5, 4, 3, 2, 1, or no nitrogen-containing groups or nitrogen atoms in the side groups. 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 units of Formula I are located in the main chain of polycarbonate rather than side groups.

[0105] 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.

[0106] In some embodiments, the protonable nitrogen-containing group is a divalent group.

[0107] In some exemplary embodiments, the protonable nitrogen-containing group may include a divalent group selected from the group consisting of:

[0108] A divalent group having a protonable nitrogen atom located in the main chain and no side groups;

[0109] A divalent 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 the side;

[0110] A divalent group having a protonable nitrogen atom in the main chain and a heteroatom (e.g., one or more of O, N, S, P, Si) attached to its side.

[0111] 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.

[0112] 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.

[0113] 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.

[0114] 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.

[0115] In various specific implementations, R 3 Non-limiting examples may include:

[0116] in,

[0117] 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;

[0118] 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;

[0119] 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;

[0120] 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;

[0121] 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;

[0122] Among them, wavy lines Indicates the bonding position with adjacent atoms; and

[0123] 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.

[0124] 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.

[0125] 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.

[0126] In some embodiments, the nitrogen-containing cyclic group may have the structure of formula Ia:

[0127] in,

[0128] Y 1 It is N or C; and

[0129] 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;

[0130] 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 C1-12 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 2 They can form rings together with the ring atoms they are connected to (e.g., forming monocyclic or bicyclic rings);

[0131] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0132] 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.

[0133] In some embodiments, the nitrogen-containing cyclic group may have the structure of formula Ib:

[0134] in,

[0135] Y 2 and Y 4 Each is independently N or C, and Y 2 and Y 4 At least one of them is N;

[0136] Y 3 Selected from: bonds (such as single bonds), C, N, O, and S;

[0137] Y 5 Selected from: bonds (such as single bonds), C, N, O, and S;

[0138] Each occurrence of n is an integer between 0 and 2.

[0139] 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;

[0140] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0141] In some implementations, ring R BIt 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.

[0142] In some exemplary embodiments, the nitrogen-containing cyclic group may include the following divalent groups:

[0143] 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;

[0144] 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;

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

[0146] 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.

[0147] 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).

[0148] 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.

[0149] In some cases, the polycarbonate described herein may also contain one or more additional groups selected from the group consisting of redox-sensitive groups, hydrocarbon groups (e.g., saturated or unsaturated, substituted or unsubstituted chain hydrocarbon groups or cyclic hydrocarbon groups), oxygen-containing groups (e.g., ether groups, epoxy groups, ketone groups, etc.), and combinations thereof. The one or more additional groups may be monovalent or divalent groups.

[0150] In some embodiments, the one or more additional groups may be located on the main chain and / or side groups of the polycarbonate.

[0151] In some embodiments, the one or more additional groups may be located in the nitrogen-containing repeating unit of the polycarbonate (e.g., the repeating unit of Formula I) or other repeating units.

[0152] The polycarbonate described herein may also have at least one (e.g., 1, 2, 3, 4, or 5) redox-sensitive groups in its nitrogen-containing repeating unit (e.g., a repeating unit of Formula I, such as structure A) or other repeating units. In some embodiments, the redox-sensitive groups include oxidation-sensitive and / or reduction-sensitive groups. In some embodiments, the redox-sensitive groups include reduction-sensitive groups.

[0153] 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).

[0154] 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.

[0155] The polycarbonate described herein may also have one or more (e.g., 1, 2, 3, 4, 5 or 6) hydrocarbon groups in its nitrogen-containing repeating unit (e.g., repeating unit of Formula I, such as structure A) or other repeating unit.

[0156] Such hydrocarbon groups may include, for example, saturated or unsaturated, substituted or unsubstituted chain hydrocarbon groups or cyclic hydrocarbon groups. In some embodiments, the hydrocarbon group may include unsubstituted or substituted C-type hydrocarbons. 1-24 (For example, C) 1-20 C 1-16 Or C 1-12 ) hydrocarbon group (e.g., saturated or unsaturated, straight-chain or cyclic aliphatic or aromatic hydrocarbon group), wherein the substituent can be selected from hydrogen, deuterium, tritium, hydroxyl, halogen (e.g., fluorine, chlorine, bromine, iodine), carboxyl, nitro, amino, cyano, ether, substituted or unsubstituted C 1-12 Ester group, 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 and substituted or unsubstituted 5- to 14-membered heteroaryl.

[0157] The one or more hydrocarbon groups may be located on the main chain and / or side groups of the polycarbonate. For example, at least one hydrocarbon group is located on the main chain of the polycarbonate. In some embodiments, in the repeating unit of Formula I, the number of hydrocarbon groups located on the main chain of the polycarbonate may be greater than the number of hydrocarbon groups located on the side groups.

[0158] The polycarbonate described herein may also have one or more (e.g., 1, 2, 3, 4, 5 or 6) oxygen-containing groups in its nitrogen-containing repeating unit (e.g., repeating unit of Formula I, such as structure A).

[0159] Such oxygen-containing groups may include, for example, ether groups, epoxy groups, ketone groups, etc.

[0160] The one or more oxygen-containing groups may be located in the main chain and / or side groups of the polycarbonate. For example, at least one oxygen-containing group is located in the main chain of the polycarbonate. In some embodiments, such as in a repeating unit of Formula I, the number of oxygen-containing groups located in the main chain of the polycarbonate may be greater than the number of oxygen-containing groups located in the side groups.

[0161] In the repeating unit of Formula I, any two of the at least one nitrogen-containing group and at least one additional group are interconnected via L I Connected, where L I It is a bond (such as a single bond), a carbonate bond, or a C bond. 1-16 Hydroxyl group.

[0162] 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.

[0163] In some cases, the polycarbonate or its salt may also contain one or more repeating units of formula V:

[0164] in,

[0165] A x It contains at least one (e.g., at least 1, 2, 3, 4, 5 or 6) group selected from the group consisting of nitrogen-containing groups, redox-sensitive groups, hydrocarbon groups, oxygen-containing groups, and combinations thereof as described above;

[0166] R 1Each 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;

[0167] a3 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 within a range formed by any two of the above values ​​as endpoints;

[0168] b3 and b4 are each an integer from 0 to 2, for example, 0, 1 or 2.

[0169] Generally, when the polycarbonate contains repeating units of formula V, the repeating units of formula V are different from the repeating units of formula I.

[0170] The nitrogen-containing group, redox-sensitive group, hydrocarbon group, and oxygen-containing group can be divalent groups.

[0171] In the repeating unit of Formula V, at least one group selected from the nitrogen-containing group, redox-sensitive group, hydrocarbon group, oxygen-containing group, and combinations thereof may be located on the main chain and / or side group of the polycarbonate, for example, on the main chain. In some embodiments, the number of at least one group selected from the nitrogen-containing group, redox-sensitive group, hydrocarbon group, oxygen-containing group, and combinations thereof located on the main chain of the polycarbonate may be greater than the number located on the side group. In some embodiments, the repeating unit of Formula V contains no more than 4, 3, 2, 1, or no at least one group selected from the nitrogen-containing group, redox-sensitive group, hydrocarbon group, oxygen-containing group, and combinations thereof on the side group. In some embodiments, in the repeating unit of Formula V, some or all of at least one group selected from the nitrogen-containing group, redox-sensitive group, hydrocarbon group, oxygen-containing group, and combinations thereof are located on the main chain of the polycarbonate. In some embodiments, in the repeating units of Formula V, at least 50%, 60%, 70%, 80%, 90%, or 100% of the group selected from the nitrogen-containing group, redox-sensitive group, hydrocarbon group, oxygen-containing group, and combinations thereof are located on the main chain of the polycarbonate rather than on the side groups.

[0172] In the repeating unit of formula V, any two of the groups selected from at least one of the nitrogen-containing group, redox-sensitive group, hydrocarbon group, and oxygen-containing group are transmitted via L V Connected, where L V It is a bond (such as a single bond), a carbonate bond, or a C bond. 1-16 Hydroxyl group.

[0173] In the polycarbonate, the degree of polymerization of the repeating unit of formula V 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 V can be 2-1500, 2-1000, 3-800, 3-500, 3-200, 3-150 or 3-100.

[0174] In some implementations, repeating units can be directly bonded together.

[0175] In some exemplary cases, the polycarbonate or a salt thereof may have repeating units of formula VI:

[0176] in,

[0177] 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;

[0178] L represents a key (such as a single key), C represents a bond. 1-16 (For example, C) 1-12 C 1-10 C 1-8 C 1-6 C 1-4 Or C 1-3 ) Hydroxyl groups (e.g., alkylene or alkenylene) or carbonate bonds;

[0179] 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 1-12Alkylamino, 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;

[0180] p1 to p4 are each an independent integer from 0 to 2, for example, 0, 1 or 2;

[0181] q1 and q4 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

[0182] q2 and q3 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 within a range formed by any two of the above values ​​as endpoints.

[0183] In such polycarbonate, the degree of polymerization of the repeating unit of Formula VI 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 embodiments, the degree of polymerization of the repeating units of Formula VI can be 2-1500, 2-1000, 3-800, 3-500, 3-200, 3-150, or 3-100. In some embodiments, the repeating units of Formula VI are directly bonded to each other.

[0184] In Formula VI, 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.

[0185] In some embodiments, the repeating unit of Formula VI has one or more nitrogen-containing groups. These 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. 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 VI 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 VI 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 VI are located in the main chain of the polycarbonate rather than in the side groups.

[0186] In some embodiments, the polycarbonate is protonable by introducing at least one protonable nitrogen-containing group as described herein into the repeating unit of Formula VI.

[0187] In some embodiments, 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, in the repeating unit of Formula VI, the number of redox-sensitive groups located on the main chain of the polycarbonate may be greater than the number of redox-sensitive groups located on the side groups. In some embodiments, the repeating unit of Formula VI 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 VI are located on the main chain of the polycarbonate. In some embodiments, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or 100% of the redox-sensitive groups in the repeating unit of Formula VI are located on the main chain of the polycarbonate rather than on the side groups.

[0188] The repeating unit of Formula VI may comprise one or more nitrogen-containing groups and / or one or more redox-sensitive groups. 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. 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).

[0189] 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.

[0190] In some cases, the polycarbonate may be a linear polymer. In some embodiments, the polycarbonate is not a crosslinked polymer. In some specific embodiments, the polycarbonate may or may not be a random copolymer.

[0191] 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.

[0192] The polycarbonate may have a weight-average molecular weight (Mn) of 0.5 to 100 kDa. wFor 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 .

[0193] 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.

[0194] In some embodiments, the polycarbonate may also be present in its salt form. For example, the salt of the polycarbonate is formed by salting with anions such as chloride, acetate, citrate, tetrafluoroacetate, etc.

[0195] Preparation of polycarbonate

[0196] On the other hand, this article provides a method for preparing polycarbonate, comprising a polymerization reaction using at least one monomer having formula II and at least one monomer having formula III:

[0197] In Equations II and III,

[0198] A 1 and A 2 Each independently comprises one or more groups selected from the group consisting of formula IV:

[0199] in,

[0200] M is selected from: nitrogen-containing groups, redox-sensitive groups, hydrocarbon groups, oxygen-containing groups, and combinations thereof;

[0201] x1 and x2 are each an integer from 0 to 2, for example, 0, 1 or 2;

[0202] y1 and y2 are each 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;

[0203] R1 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;

[0204] Among them, A 1 and A 2 At least one of them has a nitrogen-containing group;

[0205] Among them, for A 1 and A 2 In the case of two or more groups that independently have Formula IV, the two or more groups that independently have Formula IV are connected to each other via L M Connected, where L M It is a bond, carbonate bond or C 1-16 Hydroxyl group; and

[0206] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0207] For the sake of brevity, for a detailed description of the nitrogen-containing groups, redox-sensitive groups, hydrocarbon groups, and oxygen-containing groups contained in the monomers used in the above methods, please refer to the relevant description of polycarbonate above.

[0208] In some implementations, A 1 It contains a group of formula IV. In some embodiments, A 2 It contains a group having formula IV.

[0209] In other exemplary embodiments, the polycarbonate may be prepared by a method comprising polymerizing a monomer having formula VII with a monomer having formula VIII:

[0210] in,

[0211] X 1 and X 2 Each independently is: a key (such as a single key) or

[0212] Y1 and Y 2 Each independently is: a key (such as a single key) or

[0213] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group;

[0214] L represents a key (such as a single key), C represents a bond. 1-16 (For example, C) 1-12 C 1-10 C 1-8 C 1-6 C 1-4 Or C 1-3 ) Hydroxyl groups (e.g., alkylene or alkenylene) or carbonate bonds;

[0215] 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;

[0216] p1 to p4 are each an independent integer from 0 to 2, for example, 0, 1 or 2;

[0217] q1 and q4 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

[0218] q2 and q3 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 within a range formed by any two of the above values ​​as endpoints;

[0219] The restrictions are:

[0220] (1) Equation VII and Equation VIII 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 VII or Equation VIII at the same time;

[0221] (2) At least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0222] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0223] 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.

[0224] 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. In some implementations, -X 1 -Y 1 -Can be used with -X 2 -Y 2 - Same or different.

[0225] 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.

[0226] In another exemplary implementation, X 1 yes And Y 1 yes X 2 yes Or Y 2 yes And L is a bond; where X 1 Can be with X 2 Same or different, Y 1 Can be with Y 2 Same or different.

[0227] In another exemplary implementation, X 1 yes Or Y 1 yes X 2 yes And Y 2 yes And L is a bond; where X 1 Can be with X 2 Same or different, Y 1 Can be with Y 2 Same or different.

[0228] The above polymerization reaction can be carried out in a certain solvent system, usually an organic solvent.

[0229] 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 6-14 Aromatic 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.

[0230] 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).

[0231] 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.

[0232] 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.

[0233] 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.

[0234] 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.

[0235] On the other hand, this article also provides polycarbonates prepared by the methods described herein.

[0236] Polycarbonate composites and their preparation

[0237] On the other hand, this article provides a complex formed by polycarbonate and at least one nucleic acid according to the present article, also known as a "polycarbonate complex".

[0238] For the purposes of this document, the nucleic acid can be any nucleic acid that needs to be protected and delivered to a biological target site (e.g., cell, tissue, organ, etc.) to perform its function. In some cases, the nucleic acid includes, but is not limited to, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), peptide nucleic acid (PNA), and locked nucleic acid (LNA).

[0239] 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.

[0240] 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.

[0241] 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.

[0242] In some implementations, the nucleic acid can be endogenous or exogenous.

[0243] 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.

[0244] 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.

[0245] On the other hand, this article provides a method for preparing polycarbonate nucleic acid complexes, which includes:

[0246] - Provides polycarbonate or a salt thereof, and at least one nucleic acid; and

[0247] - Mix the polycarbonate or its salt with the at least one nucleic acid.

[0248] For the sake of brevity, for a description of the polycarbonate or its salts and the at least one nucleic acid, please refer to the “Polycarbonate” and “Polycarbonate Complex” sections above.

[0249] 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.

[0250] 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.

[0251] 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.

[0252] 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.

[0253] 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.

[0254] After mixing, the resulting mixture can be incubated for a period of time to allow for better formation of the polycarbonate-nucleic acid complex.

[0255] 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.

[0256] On the other hand, this article provides a polycarbonate composite prepared by the above preparation method.

[0257] Compositions and Products

[0258] On the other hand, this document provides a composition comprising the polycarbonate described herein, or a polycarbonate nucleic acid complex.

[0259] In some embodiments, the composition may be a pharmaceutical composition.

[0260] In some embodiments, the composition comprises a pharmaceutically acceptable carrier and at least one polycarbonate described herein.

[0261] In other embodiments, the composition comprises a pharmaceutically acceptable carrier and at least one polycarbonate complex described herein.

[0262] 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.

[0263] 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.

[0264] It should be understood that the relative amounts of the active ingredient, pharmaceutically acceptable carrier, and / or other components in the composition are variable and may depend on various factors, such as the age, sex, weight, physical condition, and route of administration of the target recipient, depending on the application of the active ingredient (such as polycarbonate or its complexes as described herein). As an example, the composition may contain 0.1% to 100% (w / w) of the active ingredient (such as polycarbonate nucleic acid complexes as 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 polycarbonate nucleic acid complexes as described herein) formed by any two of the above values ​​as endpoints.

[0265] 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.

[0266] In some embodiments, the composition may also contain other active substances.

[0267] In some embodiments, the composition is formulated as an injection, oral preparation, topical preparation, inhaler, or implant, such as a solution, emulsion, suspension, gel, ointment, patch, capsule, tablet, powder inhaler, aerosol, or other dosage form.

[0268] On the other hand, this article provides a product comprising:

[0269] -The polycarbonate composites described herein;

[0270] - A composition comprising the polycarbonate composite described herein; or

[0271] - The polycarbonate or its salt and at least one nucleic acid described herein.

[0272] 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.

[0273] 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.

[0274] 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.

[0275] 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).

[0276] 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.

[0277] Usage and Purpose

[0278] On the other hand, a method for delivering at least one nucleic acid to a cell or object is provided, the method comprising:

[0279] - Provides a complex as described herein, said complex being formed by combining said at least one nucleic acid with a polycarbonate or a salt thereof as described herein; and

[0280] - To bring the cell or the object into contact with the complex.

[0281] On the other hand, a method for expressing at least one nucleic acid in a cell or object is provided, the method comprising:

[0282] - Provides a complex as described herein, said complex being formed by combining said at least one nucleic acid with a polycarbonate or a salt thereof as described herein; and

[0283] - To bring the cell or the object into contact with the complex.

[0284] 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.

[0285] 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.

[0286] 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.

[0287] 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.

[0288] 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.

[0289] 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.

[0290] 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.

[0291] 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.

[0292] 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.

[0293] In some embodiments, the delivery or expression is in vitro or in vivo.

[0294] In some embodiments, the delivery may be carried out via intra-arterial, intravenous, intraperitoneal, extra-gastric, intramuscular, subcutaneous, oral, inhalation, or local routes.

[0295] The development of polycarbonate described in this article opens a new pathway for the biodelivery of nucleic acids. Effective delivery of nucleic acids to biological target sites (such as within cells) has always been a significant challenge, as nucleic acids are not only easily inactivated or degraded by cationic interference during delivery, but also need to overcome the cell membrane barrier to enter the cell nucleus. The polycarbonate nanocarriers described in this article can carry a wide range of nucleic acids of various molecular weights and lengths, exhibiting extremely high biodelivery adaptability. As a carrier, the polycarbonate provides effective protection, promotes transmembrane transport, and degrades promptly upon arrival at the specific environment (such as the biological target site), 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.

[0296] 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.

[0297] Implementation methods / combinations

[0298] Embodiment A1: A polycarbonate or a salt thereof, said polycarbonate having repeating units of Formula I:

[0299] in,

[0300] A contains at least one nitrogen-containing group;

[0301] 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;

[0302] a1 and a2 are each independent integers from 1 to 16;

[0303] b1 and b2 are each an independent integer between 0 and 2.

[0304] Embodiment A2: The polycarbonate or its salt according to Embodiment A1, wherein the at least one nitrogen-containing group comprises at least one protonable nitrogen-containing group.

[0305] Embodiment A3: The polycarbonate or its salt according to Embodiment A1, wherein the at least one nitrogen-containing group is located in the main chain of the polycarbonate.

[0306] Embodiment A4: 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 of the polycarbonate.

[0307] Embodiment A5: The polycarbonate or its salt according to Embodiment A2, wherein the at least one protonable nitrogen-containing group is located in the main chain of the polycarbonate.

[0308] Embodiment A6: The polycarbonate or its salt according to Embodiment A2, wherein at least one nitrogen atom in the at least one protonable nitrogen-containing group is located in the main chain of the polycarbonate.

[0309] Embodiment A7: The polycarbonate or its salt according to Embodiment A2, 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.

[0310] Embodiment A8: The polycarbonate or its salt according to Embodiment A7, wherein the amino group is selected from the group consisting of secondary amino groups, tertiary amino groups, quaternary amino groups, and combinations thereof.

[0311] Embodiment A9: The polycarbonate or its salt according to Embodiment A7, 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 having at least one cyclic nitrogen atom, either fused together with each other or with an aliphatic or aromatic carbon ring, and combinations thereof.

[0312] Embodiment A10: The polycarbonate or salt thereof according to Embodiment A1, wherein the polycarbonate further comprises one or more additional groups selected from the group consisting of redox-sensitive groups, hydrocarbon groups, oxygen-containing groups, and combinations thereof.

[0313] Embodiment A11: The polycarbonate or salt thereof according to Embodiment A10, wherein the one or more additional groups are located in the main chain and / or side groups of the polycarbonate.

[0314] Embodiment A12: The polycarbonate or salt thereof according to Embodiment A10, wherein the one or more additional groups are located in the main chain of the polycarbonate.

[0315] Embodiment A13: The polycarbonate or salt thereof according to Embodiment A10, wherein the one or more additional groups are located in the repeating unit of Formula I and / or other repeating units.

[0316] Embodiment A14: The polycarbonate or salt thereof according to Embodiment A10, wherein the polycarbonate contains at least one redox-sensitive group.

[0317] Embodiment A15: The polycarbonate or its salt according to Embodiment A14, 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.

[0318] Embodiment A16: The polycarbonate or salt thereof according to Embodiment A10, wherein the polycarbonate contains at least one hydrocarbon group.

[0319] Embodiment A17: The polycarbonate or salt thereof according to Embodiment A16, wherein the at least one hydrocarbon group comprises one or more of saturated or unsaturated, substituted or unsubstituted chain hydrocarbon groups and cyclic hydrocarbon groups.

[0320] Embodiment A18: The polycarbonate or salt thereof according to Embodiment A10, wherein the polycarbonate contains at least one oxygen-containing group.

[0321] Embodiment A19: The polycarbonate or its salt according to Embodiment A18, wherein the at least one oxygen-containing group comprises one or more of ether group, epoxy group, and ketone group.

[0322] Embodiment A20: The polycarbonate or a salt thereof according to Embodiment A1, wherein the polycarbonate further comprises one or more repeating units of formula V:

[0323] in,

[0324] A x It contains at least one group selected from the group consisting of nitrogen-containing groups, redox-sensitive groups, hydrocarbon groups, oxygen-containing groups, and combinations thereof, as described above.

[0325] 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 1-12Alkyl 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;

[0326] a3 and a4 are each independent integers from 1 to 16;

[0327] b3 and b4 are each an independent integer from 0 to 2.

[0328] Embodiment A21: The polycarbonate or a salt thereof according to Embodiment A1, wherein the polycarbonate has a repeating unit of Formula VI:

[0329] in,

[0330] 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;

[0331] L is a single bond, C 1-16 Hydroxyl or carbonate bond;

[0332] 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;

[0333] p1 to p4 are each an independent integer from 0 to 2;

[0334] q1 and q4 are each independent integers from 1 to 16; and

[0335] q2 and q3 are each independent integers from 0 to 16.

[0336] Embodiment A22: The polycarbonate or a salt thereof according to Embodiment A1, wherein the polycarbonate has a weight-average molecular weight (M) of 0.5 to 100 kDa. w ).

[0337] Embodiment A23: The polycarbonate or salt thereof according to Embodiment A1, wherein the polycarbonate has a pKa of 3.5-9.5.

[0338] Implementation Method B1: A method for preparing polycarbonate, the method comprising:

[0339] Polymerize at least one monomer having Formula II with at least one monomer having Formula III:

[0340] In Equations II and III,

[0341] A 1 and A 2 Each independently comprises one or more groups selected from the group consisting of formula IV:

[0342] in,

[0343] M is selected from: nitrogen-containing groups, redox-sensitive groups, hydrocarbon groups, oxygen-containing groups, and combinations thereof;

[0344] x1 and x2 are each an independent integer from 0 to 2;

[0345] y1 and y2 are each independent integers from 0 to 16;

[0346] 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;

[0347] Among them, A 1 and A 2 At least one of them has a nitrogen-containing group;

[0348] Among them, for A 1 and A 2 In the case of two or more groups that independently have Formula IV, the two or more groups that independently have Formula IV are connected to each other via L M Connected, where L M It is a bond, carbonate bond or C1-16 Hydroxyl group; and

[0349] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0350] Implementation method B2: According to the method of implementation method B1, wherein A 1 It contains a group having formula IV.

[0351] Implementation method B3: According to the method described in implementation method B1, wherein A 2 It contains a group having formula IV.

[0352] Implementation Method B4: The method according to Implementation Method B1, wherein the method includes:

[0353] The monomer having formula VII is subjected to a polymerization reaction with a monomer having formula VIII:

[0354] in,

[0355] X 1 and X 2 Each independently is: key or

[0356] Y 1 and Y 2 Each independently is: key or

[0357] X and Y each independently contain at least one nitrogen-containing group and / or at least one redox-sensitive group;

[0358] L is a key, C 1-16 Hydroxyl or carbonate bond;

[0359] 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;

[0360] p1 to p4 are each an independent integer from 0 to 2;

[0361] q1 and q4 are each independent integers from 1 to 16; and

[0362] q2 and q3 are each independent integers from 0 to 16;

[0363] The restrictions are:

[0364] (1) Equation VII and Equation VIII 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 VII or Equation VIII at the same time;

[0365] (2) At least one of X and Y contains at least one protonable nitrogen-containing group, and X is different from Y;

[0366] Among them, wavy lines It indicates the bonding position with adjacent atoms.

[0367] Implementation C1: A complex of polycarbonate or a salt thereof according to Implementation A1 with at least one nucleic acid.

[0368] 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.

[0369] 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.

[0370] 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.

[0371] Embodiment C5: The polycarbonate nucleic acid complex according to Embodiment C1, wherein the complex has an average particle size between 10 and 1000 nm.

[0372] Embodiment D1: A composition comprising the polycarbonate or a salt thereof as described in Embodiment A1, or the complex as described in Embodiment C1.

[0373] Implementation D2: The composition according to Implementation D1, wherein the composition is a pharmaceutical composition.

[0374] Implementation D3: The composition according to Implementation D2 further comprises at least one pharmaceutically acceptable carrier.

[0375] 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.

[0376] 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.

[0377] 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.

[0378] Embodiment D7: The composition according to Embodiment D1, wherein the composition further comprises other active substances.

[0379] 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.

[0380] Implementation method E1: A product comprising:

[0381] -The polycarbonate or its salt thereof and at least one nucleic acid according to embodiment A1;

[0382] -The complex according to embodiment C1; or

[0383] -The composition according to embodiment D1.

[0384] 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.

[0385] 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.

[0386] 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.

[0387] Implementation F1: A method for delivering at least one nucleic acid to a cell or object, the method comprising:

[0388] - 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

[0389] - To bring the cell or the object into contact with the complex.

[0390] Implementation method G1: A method for expressing at least one nucleic acid in a cell or object, the method comprising:

[0391] - 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

[0392] - To bring the cell or the object into contact with the complex.

[0393] 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.

[0394] Embodiment I1: 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.

[0395] Embodiment J1: 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.

[0396] 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.

[0397] 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.

[0398] 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.

[0399] 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.

[0400] 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.

[0401] 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.

[0402] 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.

[0403] 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 an intra-arterial, intravenous, intraperitoneal, extra-gastric, intramuscular, subcutaneous, oral, local or inhalation route.

[0404] Example

[0405] 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.

[0406] Materials and Instruments

[0407] The instrument information used in the embodiments is summarized in Table 1 below:

[0408] Table 1. Instruments used in the examples

[0409] The material information used in the embodiments is summarized in Table 2 below:

[0410] Table 2. Materials used in the examples

[0411] Example 1: Monomer Preparation and Characterization

[0412] Preparation Example A1: 2,2'-((3-methoxypropyl)azadiyl)bis(1-ethanol) (A013)

[0413] 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 Na₂SO₄. The solvent was removed by vacuum distillation, and the residue was dried to obtain monomer A013 (1.20 g), yield 71.1%.

[0414] 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).

[0415] HRMS(ESI,m / z)C8H 19 NO3, [M+H] + Calculated value: 178.1443; Measured value: 178.1428.

[0416] Preparation Example A2: 2,2'-((3-(pyrrolidone-1-yl)propyl)azadiyl)bis(1-ethanol)(A014)

[0417] 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%.

[0418] 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).

[0419] HRMS(ESI,m / z):C 11 H 24 N₂O₂,[M+H]⁺ Calculated value: 217.1916; Measured value: 217.1935.

[0420] Preparation Example A3: 2,2'-((3-(piperidin-1-yl)propyl)azadiyl)bis(1-ethanol) (A015)

[0421] 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%.

[0422] 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).

[0423] HRMS(ESI,m / z):C 12 H 26 Calculated value of N2O2,[M+H]+: 231.2073; Measured value: 231.2083.

[0424] Preparation Example A4: 2,2'-((3-morpholinopropyl)azadiyl)bis(1-ethanol) (A016)

[0425] 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%.

[0426] 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).

[0427] HRMS(ESI,m / z):C 11 H 24 N₂O₃,[M+H]⁺ Calculated value: 233.1865; Measured value: 233.1883.

[0428] Preparation Example A5: 2,2'-((3-(1H-pyrrolo-1-yl)propyl)azadiyl)bis(1-ethanol) (A017)

[0429] 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%.

[0430] 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).

[0431] HRMS(ESI,m / z):C 11 H 20 Calculated value of N2O2,[M+H]+: 213.1603; Measured value: 213.1612.

[0432] Preparation Example A6: 2,2'-((3-(4-methylpiperazin-1-yl)propyl)azadiyl)bis(1-ethanol) (A018)

[0433] 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.

[0434] 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).

[0435] HRMS(ESI,m / z):C 12 H 27 Calculated value of N3O2,[M+H]+: 246.2182; Measured value: 246.2196.

[0436] Preparation Example A7: 2,2'-((3-(dimethylamino)propyl)azadiyl)bis(1-ethanol) (A019)

[0437] 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%.

[0438] 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).

[0439] HRMS(ESI,m / z):C9H 22 Calculated value of N2O2,[M+H]+: 191.1760; Measured value: 191.1749.

[0440] Preparation Example A8: 2,2'-Thiobis(1-ethanol) (A025)

[0441] 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 Chromatographic 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.

[0442] 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).

[0443] Preparation Example A9: 2,2'-trithionide dimethylbis(1-ethanol) (A027)

[0444] 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 (15.0 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.

[0445] 1 H NMR (700MHz, CDCl3): δ2.95(t,SCH2CH2OH), 4.05(q,SCH2CH2OH), 3.15(t,SCH2CH2OH).

[0446] HRMS(ESI,m / z):C7H 12 O4S2,[M+Na] + Calculated value: 247.0075; Measured value: 247.0103.

[0447] Preparation Example A10: 2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethane-1-ol))(A028)

[0448] (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%.

[0449] 1 H NMR (700MHz, DMSO-d6): δ12.63(s,CH2COOH),3.37(s,SCH2COOH),1.54(s,(CH3)2CSS).

[0450] HRMS(ESI,m / z):C7H 12 O4S2,[M+Na] + Calculated value: 247.0075; Measured value: 247.0103.

[0451] (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. The mixture was then extracted with diethyl ether (3×, 500 mL), and the organic layer was collected and dried with anhydrous Na2SO4. 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%.

[0452] 1 H NMR (700MHz, DMSO-d6) δ4.82(t,SCH2CH2OH),3.52(q,SCH2CH2OH),2.65(t,SCH2CH2OH),1.53(s,(CH3)2CSS).

[0453] HRMS(ESI,m / z):C7H 16 O2S2,[M+Na] + Calculated value: 219.0489; Measured value: 219.0512.

[0454] Preparation Example A11: 3,3'-(propane-2,2-diylbis(thioalkyldiyl))bis(prop-1-ol) (A029)

[0455] (1) A similar process to that described in Preparation Example A8 was used, except that thioglycolic acid in (1) was replaced with 3-mercaptopropionic acid. Additionally, the intermediate added in step (2) was synthesized in Example (1). The yields of the intermediate and monomer A029 obtained after post-treatment were 61.5% and 94.0%, respectively.

[0456] Intermediate:

[0457] 1 H NMR (700MHz, DMSO-d6): δ12.26(s,CH2COOH),2.72(t,SCH2CH2COOH),2.48(t,SCH2CH2COOH),1.51(s,(CH3)2CSS).

[0458] HRMS(ESI,m / z):C9H 16 O4S 2, [M+Na] + Calculated value: 275.0388; Measured value: 275.0381.

[0459] Monomer A029:

[0460] 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).

[0461] HRMS(ESI,m / z):C9H 20 O2S2,[M+Na] + Calculated value: 247.0802; Measured value: 247.0799.

[0462] Preparation Example A12: 3-((2-hydroxyethyl)(methyl)amino)prop-1-ol (A084)

[0463] Weigh 2.50 g (28.05 mmol) of 3-(aminomethyl)-1-propanol, 7.01 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 A084 (1.62 g) was obtained in a yield of 43.3%.

[0464] 1 H 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).

[0465] HRMS(ESI,m / z):C6H 16NO2; [M+H]+ Calculated value: 134.1181; Measured value: 134.1175.

[0466] Preparation Example A13: 5-((3-hydroxypropyl)(methyl)amino)pentan-1-ol (A086)

[0467] 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%.

[0468] 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).

[0469] HRMS(ESI,m / z):C9H 22 NO2; [M+H]+ Calculated value: 176.1651; Measured value: 176.1654.

[0470] Preparation Example A14: 6-((3-hydroxypropyl)(methyl)amino)hexane-1-ol (A087)

[0471] Weigh 3-(aminomethyl)-1-propanol (3.500 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%.

[0472] 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).

[0473] HRMS(ESI,m / z):C 10 H 24 NO2; [M+H] + Calculated value: 190.1807; Measured value: 190.1811.

[0474] Preparation Example A15: 7-((3-hydroxypropyl)(methyl)amino)hepta-1-ol (A088)

[0475] 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%.

[0476] 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).

[0477] HRMS(ESI,m / z):C 11 H 26 NO2; [M+H] + Calculated value: 204.1964; Measured value: 204.1969.

[0478] Preparation Example A16: 5-(3-(hydroxymethyl)piperidin-1-yl)pentan-1-ol (A131)

[0479] 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 a yield of 55.1%.

[0480] 1 H 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).

[0481] HRMS(ESI,m / z):C 11 H 24 NO2; [M+H] + Calculated value: 202.1807; Measured value: 202.1810.

[0482] Preparation Example A17: 4-(Ethyl(2-hydroxyethyl)amino)but-1-ol (A089)

[0483] Weigh 2.000 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%.

[0484] 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).

[0485] HRMS(ESI,m / z):C8H 20 NO2; [M+H] + Calculated value: 162.1494; Measured value: 162.1498.

[0486] Preparation Example A18: 6-((4-hydroxybutyl)(isopropyl)amino)hexyl-1-ol (A105)

[0487] Weigh 2.50 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.

[0488] 1H NMR(400MHz, CDCl3): δ3.65(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),3.14(t, HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),2.48(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH 3)2)CH2CH2CH2CH2OH),1.57(m,HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.40(m, HOCH2CH2CH2CH2CH2CH2N(CH(CH3)2)CH2CH2CH2CH2OH),1.06(s,HOCH2CH2CH2CH2N(CH(CH3)2)).

[0489] HRMS(ESI,m / z):C 13 H 30 NO2; [M+H] + Calculated value: 232.2277; Measured value: 232.2281.

[0490] Preparation Example A19: 7-((4-hydroxybutyl)(isopropyl)amino)hepta-1-ol (A106)

[0491] 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.

[0492] 1H 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)).

[0493] HRMS(ESI,m / z):C 14 H 32 NO2; [M+H] + Calculated value: 246.2433; Measured value: 246.2438.

[0494] Preparation Example A20: 5-((2-hydroxyethyl)(isopropyl)amino)pentan-1-ol (A107)

[0495] 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.

[0496] 1H 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)).

[0497] HRMS(ESI,m / z):C 10 H 24 NO2; [M+H] + Calculated value: 190.1807; Measured value: 190.1811.

[0498] Preparation Example A21: 3,3'-(piperazine-1,4-diyl)bis(propane-1-ol) (A002)

[0499] 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%.

[0500] 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).

[0501] HRMS(ESI,m / z):C 10 H 22 N₂O₂; [M+H] +Calculated value: 203.1760; Measured value: 203.1761.

[0502] Preparation Example A22: 3-(4-(2-hydroxyethyl)piperidin-1-yl)prop-1-ol (A070)

[0503] 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%.

[0504] 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).

[0505] HRMS(ESI,m / z):C 10 H 21 NO2; [M+H] + Calculated value: 188.1651; Measured value: 188.1653.

[0506] Preparation Example A23: 3,3'-(1,4-diazacycloheptane-1,4-diyl)bis(prop-1-ol) (A073)

[0507] 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... 18After column chromatography purification (eluting agents were acetonitrile and 0.5% ammonia), monomer A073 (1.16 g) was obtained in a yield of 26.86%.

[0508] 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).

[0509] HRMS(ESI,m / z):C 11 H 24 N₂O₂; [M+H] + Calculated value: 217.1916; Measured value: 217.1917.

[0510] Preparation Example A24: 3-(3-(hydroxymethyl)pyrrolidone-1-yl)prop-1-ol (A074)

[0511] 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%.

[0512] 1 H NMR (400MHz, CDCl3): δ3.77(t,HOCH2CH2CH2N),3.54(dd,HOCH2CH(CH2)CH2CH2),3.47(dd,HOCH2CH(CH2)CH2CH2),2.79-2.29(m,H OCH2CH2CH2NCH2CH2),1.94(m,HOCH2CH(CH2)CH2CH2NCH2CH2),1.71(dd,HOCH2CH2CH2N),1.49(m,HOCH2CH(CH2)CH2CH2NCH2CH2).

[0513] HRMS(ESI,m / z):C8H 17 NO2; [M+H] + Calculated value: 160.1338; Measured value: 160.1339.

[0514] Preparation Example A25: 2,2'-(piperidine-1,2-diyl)bis(ethanol) (A071)

[0515] 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%.

[0516] 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).

[0517] HRMS(ESI,m / z): C9H 19 NO2; [M+H] + Calculated value: 174.1494; Measured value: 174.1484.

[0518] Preparation Example A26: 3-(2-(2-hydroxyethyl)piperidin-1-yl)prop-1-ol (A082)

[0519] 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%.

[0520] 1H 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).

[0521] HRMS(ESI,m / z): C 10 H 21 NO2; [M+H] + Calculated value: 188.1651; Measured value: 188.1654.

[0522] Preparation Example A27: 3-(3-(hydroxymethyl)piperidin-1-yl)propanol (A075)

[0523] 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 A075 (0.93 g) was obtained in a yield of 62%.

[0524] 1 H NMR (400MHz, CDCl3): δ3.77(t,CHCH2OH),3.54-3.42(m,NCH2CH2CH2OH),2.98(d,CHCH2OH),2.83(s,CHCH2OH),2.57 (m,NCH2CH2CH2CHCH2,NCH2CH2CH2CH),2.01(m,NCH2CH2CH2OH),1.91-1.52(m,NCH2CH2CH2CHCH2,NCH2CH2CH2OH),.

[0525] HRMS(ESI,m / z):C9H 19 NO2; [M+H] + Calculated value: 174.1494; Measured value: 174.1498

[0526] Preparation Example A28: 2,2'-(piperidine-1,4-diyl)bis(1-ethanol) (A078)

[0527] 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%.

[0528] 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,HOCH2CH2CH(CH2 CH2)CH2CH2),1.70(d,HOCH2CH2N(CH2CH2)CH2CH2),1.52(q,HOCH2CH2CH),1.45(dd,HOCH2CH2CH),1.25(qd,HOCH2CH2CH(CH2CH2)CH2CH2).HRMS(ESI,m / z):C9H 19 NO2; [M+H] + Calculated value: 174.1494; Measured value: 174.1495.

[0529] Preparation Example A29: 3-((2-hydroxyethyl)(isopropyl)amino)prop-1-ol (A095)

[0530] 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%.

[0531] 1H 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).

[0532] HRMS(ESI,m / z):C8H 19 NO2; [M+H] + Calculated value: 162.1494; Measured value: 162.1499.

[0533] Preparation Example A30: 3,3'-(isopropylazadiyl)bis(propan-1-ol) (A096)

[0534] 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... 18 After purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A096 (3.85 g) was obtained, with a yield of 85.7%.

[0535] 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).

[0536] HRMS(ESI,m / z):C9H 21 NO2; [M+H] + Calculated value: 176.1651; Measured value: 176.1655.

[0537] Preparation Example A31: 5-((3-hydroxypropyl)(isopropyl)amino)pentan-1-ol (A098)

[0538] 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%.

[0539] 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).

[0540] HRMS(ESI,m / z):C 11 H 25 NO2; [M+H] + Calculated value: 204.1964; Measured value: 204.1967.

[0541] Preparation Example A32: 6-((3-hydroxypropyl)(isopropyl)amino)hexane-1-ol (A099)

[0542] 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%.

[0543] 1H 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).

[0544] HRMS(ESI,m / z):C 12 H 27 NO2; [M+H] + Calculated value: 218.2120; Measured value: 218.2122.

[0545] Preparation Example A33: 7-((3-hydroxypropyl)(isopropyl)amino)heptane-1-ol (A100)

[0546] 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%.

[0547] 1 H 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).

[0548] HRMS(ESI,m / z):C 13H 29 NO2; [M+H] + Calculated value: 232.2277; Measured value: 232.2279.

[0549] Preparation Example A34: 5-(Butyl(4-hydroxybutyl)amino)pentan-1-ol (A116)

[0550] 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%.

[0551] 1 H 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)

[0552] HRMS(ESI,m / z):C 13 H 29 NO2; [M+H] + Calculated value: 232.2277; Measured value: 232.2279.

[0553] Preparation Example A35: 6-(Butyl(4-hydroxybutyl)amino)hexane-1-ol (A117)

[0554] 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%.

[0555] 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)

[0556] HRMS(ESI,m / z):C 14 H 37 NO2; [M+H] + Calculated value: 246.2433; Measured value: 246.2436.

[0557] Preparation Example A36: 7-(Butyl(4-hydroxybutyl)amino)heptane-1-ol (A118)

[0558] 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 purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A118 (3.14 g) was obtained, with a yield of 70.4%.

[0559] 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)

[0560] HRMS(ESI,m / z):C15 H 33 NO2; [M+H] + Calculated value: 260.2590; Measured value: 260.2594.

[0561] Preparation Example A37: 5,5'-(piperazine-1,4-diyl)bis(1-pentanol) (A119)

[0562] 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%.

[0563] 1 H 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).

[0564] HRMS(ESI,m / z):C 14 H 30 N₂O₂; [M+H] + Calculated value: 259.2386; Measured value: 259.2391.

[0565] Preparation Example A38: 6,6'-(piperazine-1,4-diyl)bis(hexane-1-ol) (A120)

[0566] 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%.

[0567] 1H 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).

[0568] HRMS(ESI,m / z):C 16 H 34 N₂O₂; [M+H] + Calculated value: 287.2699; Measured value: 287.2702.

[0569] Preparation Example A39: 7,7'-(piperazine-1,4-diyl)bis(heptane-1-ol) (A121)

[0570] 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%.

[0571] 1H 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).

[0572] HRMS(ESI,m / z):C 18 H 38 N₂O₂; [M+H] + Calculated value: 315.3012; Measured value: 315.3016.

[0573] Preparation Example A40: 5,5'-(1,4-diazacycloheptane-1,4-diyl)bis(1-pentan-1-ol) (A122)

[0574] 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%.

[0575] 1H 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).

[0576] HRMS(ESI,m / z):C 15 H 32 N₂O₂; [M+H] + Calculated value: 273.2542; Measured value: 273.2546.

[0577] Preparation Example A41: 6,6'-(1,4-diazacycloheptane-1,4-diyl)bis(hexane-1-ol) (A123)

[0578] 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%.

[0579] 1H 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).

[0580] HRMS(ESI,m / z):C 17 H 36 N₂O₂; [M+H] + Calculated value: 301.2855; Measured value: 301.2859.

[0581] Preparation Example A42: 7,7'-(1,4-diazacycloheptane-1,4-diyl)bis(heptane-1-ol) (A124)

[0582] 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 purification by column chromatography (eluting with acetonitrile and 0.5% ammonia), monomer A124 (4.43 g) was obtained, with a yield of 67.5%.

[0583] 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).

[0584] HRMS(ESI,m / z):C 19 H 40 N₂O₂; [M+H] + Calculated value: 329.3168; Measured value: 329.3171.

[0585] Preparation Example A43: 5-(2-(2-hydroxyethyl)piperidin-1-yl)pentan-1-ol (A128)

[0586] 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%.

[0587] 1 H 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).

[0588] HRMS(ESI,m / z):C 12 H 25 NO2; [M+H] + Calculated value: 216.1964; Measured value: 216.1967.

[0589] Preparation Example A44: 6-(2-(2-hydroxyethyl)piperidin-1-yl)hexane-1-ol (A129)

[0590] 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%.

[0591] 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).

[0592] HRMS(ESI,m / z):C 13 H 27 NO2; [M+H] + Calculated value: 230.2120; Measured value: 230.2124.

[0593] Preparation Example A45: 7-(2-(2-hydroxyethyl)piperidin-1-yl)heptane-1-ol (A130)

[0594] 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%.

[0595] 1H 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).

[0596] HRMS(ESI,m / z):C 14 H 29 NO2; [M+H] + Calculated value: 244.2277; Measured value: 244.2280.

[0597] Preparation Example A46: 6-(3-(hydroxymethyl)piperidin-1-yl)hex-1-ol (A132)

[0598] 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%.

[0599] 1 H NMR (400MHz, CDCl3): δ3.62(m,CHCH2OH,NCH2CH2CH2CH2CH2CH2OH),3.51(m,CHCH2OH),2.80(d,CHC H2OH),2.58(d,CHCH2OH),2.29(t,NCH2CH2CH2CH2CH2CH2OH),2.15(s,NCH2CH2CH2CHCH2),2.02(s,N CH2CH2CH2CHCH2),1.79(t,NCH2CH2CH2CH2CH2CH2OH),1.68(m,NCH2CH2CH2CHCH2),1.55(m,NCH2CH 2CH2CHCH2,NCH2CH2CH2CH2CH2CH2OH),1.36(m,NCH2CH2CH2CH2CH2CH2OH),1.09(m,NCH2CH2CH2CH).

[0600] HRMS(ESI,m / z):C 12 H 26 NO2 ；[M+H] + Calculated value: 216.1964; Measured value: 216.1968.

[0601] Preparation Example A47: 7-(3-(hydroxymethyl)piperidin-1-yl)hepta-1-ol (A133)

[0602] 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%.

[0603] 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,N CH2CH2CH2CHCH2),1.75(m,NCH2CH2CH2CHCH2),1.54(m,NCH2CH2CH2CHCH2,NCH2CH2CH2CH2CH2CH2CH2OH),1.32(m,NCH2CH2CH2CH2CH2CH2CH2OH),1.09(m,NCH2CH2CH2CH).

[0604] HRMS(ESI,m / z):C 13 H 28 NO2; [M+H] + Calculated value: 230.2120; Measured value: 230.2122.

[0605] Preparation Example B1: (Methylazonyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B003)

[0606] Under ice bath conditions, 20.0 g (1 eq) of A003 (N-methyldiethanolamine) was dissolved in 200 mL of DCM solvent, and 68.1 g (2.5 eq) of CDI was added. The mixture was brought 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 with anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain 40.0 g (77.6%) of yellow solid B003.

[0607] 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).

[0608] Preparation Example B2: Piperazine-1,4-diylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylate) (B004)

[0609] The process was the same as in Preparation Example B1, except that A003 was replaced with A004 (1,4-bis(2-hydroxyethyl)piperazine), resulting in B004 as a yellow oil with a yield of 96.0%.

[0610] 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).

[0611] Preparation Example B3: (Ethylazanediyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B005)

[0612] 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%.

[0613] 1H 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).

[0614] Preparation Example B4: (Butylazinediyl)bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B007)

[0615] Following the same procedure as in Preparation Example B1, except that A003 was replaced with A007 (N-butyldiethanolamine), yielding B007 as a yellow oil with a yield of 73.7%.

[0616] 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).

[0617] Preparation Example B5: 3-(1-(2-((1H-imidazol-1-carbonyl)oxy)ethyl)piperidin-4-yl)propyl 1H-imidazol-1-carboxylic acid ester (B021)

[0618] Following the same procedure as in 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.

[0619] 1 H 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).

[0620] Preparation Example B6: Thiobis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B025)

[0621] The process 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%).

[0622] 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).

[0623] Preparation Example B7: Dithiodimethylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B026)

[0624] The process was the same as in 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.

[0625] 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).

[0626] Preparation Example B8: Trithionylbis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B027)

[0627] The process was the same as in Preparation Example B1, except that A003 was replaced with A027 (2,2'-trithionide dimethylbis(1-ethanol), yielding B027 as a white solid with a yield of 55.0%).

[0628] 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).

[0629] Preparation Example B9: (propane-2,2-diylbis(thioalkyldiyl))bis(ethane-2,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B028)

[0630] The process was the same as in Preparation Example B1, except that A003 was replaced with A028(2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethanol-1-ol), yielding B028 as a yellow oil with a yield of 89.4%.

[0631] 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).

[0632] Preparation Example B10: (propane-2,2-diylbis(thioalkyldiyl))bis(propane-3,1-diyl)bis(1H-imidazol-1-carboxylic acid ester) (B029)

[0633] The process was the same as in Preparation Example B1, except that A003 was replaced with A029(2,2'-(propane-2,2-diylbis(thiodiyl))bis(ethanol-1-ol), resulting in B029 as a yellow oil with a yield of 87%.

[0634] 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).

[0635] Preparation Example B11: M008

[0636] A026 (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%.

[0637] 1H-NMR (700MHz, CDCl3): δ4.37(m,OCOCH2CH2SS), 3.05(m,OCOCH2CH2SS).

[0638] Preparation Example B12: M009

[0639] 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.

[0640] 1 H-NMR (700MHz, CDCl3): δ4.2(t,OCOCH2CH2NCH3), 2.69(t,OCOCH2CH2NCH3), 2.34(s,OCOCH2CH2NCH3).

[0641] Example 2: Polymer Preparation

[0642] The monomers used in the polymer preparation examples are summarized in Table 3 below.

[0643] Table 3. Monomers used in polymer preparation examples

[0644] Preparation Example P1: Polymer 244

[0645] Polymer 244 contains the following repeating units:

[0646] The 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.

[0647] Preparation Example P2: Polymer 245

[0648] Polymer 245 contains the following repeating units:

[0649] Following the procedure of Preparation Example P1, the raw 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.

[0650] Preparation Example P3: Polymer 246

[0651] Polymer 246 contains the following repeating units:

[0652] Following the procedure of Preparation Example P1, the raw materials were A003 (152 mg, 1 eq), B027 (507 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.

[0653] Preparation Example P4: Polymer 247

[0654] Polymer 247 contains the following repeating units:

[0655] 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.

[0656] Preparation Example P5: Polymer 250

[0657] Polymer 250 contains the following repeating units:

[0658] Following the procedure of Preparation Example P1, the raw materials were A003 (48 mg, 1 eq), B003 (148 mg, 1.2 eq), and cesium fluoride (6.0 mg, 0.1 eq), yielding a white solid 250 (105 mg, yield 80.6%). Mw: 9.0 kDa.

[0659] Preparation Example P6: Polymer 251

[0660] Polymer 251 contains the following repeating units:

[0661] Following the procedure of Preparation Example P1, 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.

[0662] Preparation Example P7: Polymer 262

[0663] Polymer 262 contains the following repeating units:

[0664] 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.

[0665] Preparation Example P8: Polymer 269

[0666] Polymer 269 contains the following repeating units:

[0667] 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.

[0668] Preparation Example P9: Polymer 273

[0669] Polymer 273 contains the following repeating units:

[0670] 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, 2 mL of ethyl acetate was added, and 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 dryness to obtain a pale yellow oil, 273 (63 mg, yield 36.1%). Mw: 3.5 Kda.

[0671] Preparation Example P10: Polymer 276

[0672] Polymer 276 contains the following repeating units:

[0673] 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.

[0674] Preparation Example P11: Polymer 281

[0675] Polymer 281 contains the following repeating units:

[0676] Following the procedure in Preparation Example P9, the raw 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.

[0677] Preparation Example P12: Polymer 282

[0678] Polymer 282 contains the following repeating units:

[0679] Following the procedure in Preparation Example P9, 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.

[0680] Preparation Example P13: Polymer 283

[0681] Polymer 283 contains the following repeating units:

[0682] Following the procedure in Preparation Example P9, the raw 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 283 (20 mg, yield 10.2%). Mw: 14.3 kDa.

[0683] Preparation Example P14: Polymer 288

[0684] Polymer 288 contains the following repeating units:

[0685] Following the procedure in Preparation Example P9, the raw materials were A005 (103 mg, 1 eq), B005 (300 mg, 1.2 eq), and cesium fluoride (11.7 mg, 0.1 eq), yielding a colorless oily product 288 (120 mg, yield 43.5%). Mw: 8.4 kDa.

[0686] Preparation Example P15: Polymer 293

[0687] Polymer 293 contains the following repeating units:

[0688] Following the procedure in Preparation Example P9, 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.

[0689] Preparation Example P16: Polymer 294

[0690] Polymer 294 contains the following repeating units:

[0691] Following the procedure in Preparation Example P9, the raw materials were A007 (100 mg, 1 eq), B007 (260 mg, 1.2 eq), and cesium fluoride (9.4 mg, 0.1 eq), yielding a pale yellow oily substance 294 (80 mg, yield 30.9%). Mw: 6.0 kDa.

[0692] Preparation Example P17: Polymer 298

[0693] Polymer 298 contains the following repeating units:

[0694] Following the procedure of Preparation Example P1, the raw 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.

[0695] Preparation Example P18: Polymer 299

[0696] Polymer 299 contains the following repeating units:

[0697] Following the procedure in Preparation Example P9, 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.

[0698] Preparation Example P19: Polymer 300

[0699] Polymer 300 contains the following repeating units:

[0700] Following the procedure in Preparation Example P9, 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.

[0701] Preparation Example P20: Polymer 301

[0702] Polymer 301 contains the following repeating units:

[0703] Following the procedure of Preparation Example P1, the raw 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.

[0704] Preparation Example P21: Polymer 303

[0705] Polymer 303 contains the following repeating units:

[0706] 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.

[0707] Preparation Example P22: Polymer 307

[0708] Polymer 307 contains the following repeating units:

[0709] Following the procedure of Preparation Example P1, the raw 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.

[0710] Preparation Example P23: Polymer 315

[0711] Polymer 315 contains the following repeating units:

[0712] Following the procedure of Preparation Example P1, the raw 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.

[0713] Preparation Example P24: Polymer 326

[0714] Polymer 326 contains the following repeating units:

[0715] Following the procedure in Preparation Example P9, 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.

[0716] Preparation Example P25: Polymer 328

[0717] Polymer 328 contains the following repeating units:

[0718] Following the procedure of Preparation Example P1, the raw 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.

[0719] Preparation Example P26: Polymer 329

[0720] Polymer 329 contains the following repeating units:

[0721] Following the procedure of Preparation Example P1, the raw 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.

[0722] Preparation Example P27: Polymer 334

[0723] Polymer 334 contains the following repeating units:

[0724] Following the procedure of Preparation Example P1, the raw materials were A038 (100 mg, 1 eq), B003 (310 mg, 1.2 eq), and cesium fluoride (12.6 mg, 0.1 eq), yielding a white crystalline solid 334 (45 mg, yield 16.5%). Mw: 1.9 kDa.

[0725] Preparation Example P28: Polymer 373

[0726] Polymer 373 contains the following repeating units:

[0727] Following the procedure of Preparation Example P1, the raw materials were A043 (51 mg, 1 eq), B004 (210 mg, 1.2 eq), and cesium fluoride (8.0 mg, 0.1 eq), yielding a white crystalline solid 373 (105 mg, yield 57.7%). Mw: 4.4 kDa.

[0728] Preparation Example P29: Polymer 376

[0729] Polymer 376 contains the following repeating units:

[0730] Following the procedure of Preparation Example P1, the raw 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.

[0731] Preparation Example P30: Polymer 379

[0732] Polymer 379 contains the following repeating units:

[0733] Following the procedure in Preparation Example P9, the starting materials were A049 (101 mg, 1 eq), B004 (166 mg, 1.2 eq), and cesium fluoride (6 mg, 0.1 eq), yielding a pale yellow oily 379 (65 mg, yield 31.8%). Mw: 2.5 kDa.

[0734] Preparation Example P31: Polymer 380

[0735] Polymer 380 contains the following repeating units:

[0736] Following the procedure of Preparation Example P1, the raw materials were A050 (101 mg, 1 eq), B004 (207 mg, 1.2 eq), and cesium fluoride (7.3 mg, 0.1 eq), yielding a white crystalline solid 380 (95 mg, yield 41.3%). Mw: 10.2 kDa.

[0737] Preparation Example P32: Polymer 381

[0738] Polymer 381 contains the following repeating units:

[0739] Following the procedure of Preparation Example P1, the raw materials were A051 (53 mg, 1 eq), B004 (250 mg, 1.2 eq), and cesium fluoride (9.0 mg, 0.1 eq), yielding a white crystalline solid 381 (70 mg, yield 33.5%). Mw: 1.9 kDa.

[0740] Preparation Example P33: Polymer 382

[0741] Polymer 382 contains the following repeating units:

[0742] Following the procedure of Preparation Example P1, the raw materials were A052 (53 mg, 1 eq), B004 (220 mg, 1.2 eq), and cesium fluoride (8.0 mg, 0.1 eq), yielding a white crystalline solid 382 (50 mg, yield 26.3%). Mw: 1.9 kDa.

[0743] Preparation Example P34: Polymer 385

[0744] Polymer 385 contains the following repeating units:

[0745] Following the procedure of Preparation Example P1, the starting materials were A055 (99 mg, 1 eq), B004 (374 mg, 1.2 eq), and cesium fluoride (13.0 mg, 0.1 eq), yielding a white solid 385 (100 mg, yield 30.1%). Mw: 1.9 kDa.

[0746] Preparation Example P35: Polymer 388

[0747] Polymer 388 contains the following repeating units:

[0748] Following the procedure of Preparation Example P1, the starting materials were A058 (102 mg, 1 eq), B004 (234 mg, 1.2 eq), and cesium fluoride (8.2 mg, 0.1 eq), yielding a white solid 388 (160 mg, yield 64.5%). Mw: 2.3 kDa.

[0749] Preparation Example P36: Polymer 389

[0750] Polymer 389 contains the following repeating units:

[0751] Following the procedure of Preparation Example P1, the raw materials were A059 (50 mg, 1 eq), B004 (210 mg, 1.2 eq), and cesium fluoride (8.0 mg, 0.1 eq), yielding a white powder solid 389 (100 mg, yield 55.2%). Mw: 3.7 kDa.

[0752] Preparation Example P37: Polymer 392

[0753] Polymer 392 contains the following repeating units:

[0754] Following the procedure of Preparation Example P1, the raw materials were A062 (94 mg, 1 eq), B004 (368 mg, 1.2 eq), and cesium fluoride (13.0 mg, 0.1 eq), yielding a yellowish-brown oily substance 392 (55 mg, yield 17.0%). Mw: 7.5 kDa.

[0755] Preparation Example P38: Polymer 394

[0756] Polymer 394 contains the following repeating units:

[0757] Following the procedure of Preparation Example P1, the raw materials were A064 (50 mg, 1 eq), B004 (210 mg, 1.2 eq), and cesium fluoride (7.0 mg, 0.1 eq), yielding a pale yellow crystalline solid 394 (60 mg, yield 33.1%). Mw: 7.7 kDa.

[0758] Preparation Example P39: Polymer 395

[0759] Polymer 395 contains the following repeating units:

[0760] Following the procedure of Preparation Example P1, the raw materials were A065 (99 mg, 1 eq), B004 (290 mg, 1.2 eq), and cesium fluoride (-10.0 mg, 0.1 eq), yielding a pale yellow crystalline solid 395 (50 mg, yield 17.9%). Mw: 2.3 kDa.

[0761] Preparation Example P40: Polymer 399

[0762] Polymer 399 contains the following repeating units:

[0763] Following the procedure of Preparation Example P1, the raw materials were A069 (100 mg, 1 eq), B004 (67 mg, 1.2 eq), and cesium fluoride (2.4 mg, 0.1 eq), yielding a pale yellow crystalline solid 399 (47 mg, yield 33.1%). Mw: 2.3 kDa.

[0764] Preparation Example P41: Polymer 431

[0765] Polymer 431 contains the following repeating units:

[0766] Following the procedure of Preparation Example P1, the raw materials were A067 (103 mg, 1 eq), B007 (180 mg, 1.2 eq), and cesium fluoride (6.4 mg, 0.1 eq), yielding a white crystalline solid 431 (120 mg, yield 56.4%). Mw: 6.8 kDa.

[0767] Preparation Example P42: Polymer 435

[0768] Polymer 435 contains the following repeating units:

[0769] Following the procedure of Preparation Example P1, the raw 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.

[0770] Preparation Example P43: Polymer 474

[0771] Polymer 474 contains the following repeating units:

[0772] 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.

[0773] Preparation Example P44: Polymer 475

[0774] Polymer 475 contains the following repeating units:

[0775] Following the procedure in Preparation Example P9, 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.

[0776] Preparation Example P45: Polymer 521

[0777] Polymer 521 contains the following repeating units:

[0778] 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 mixture was allowed to return 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 precipitate 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.

[0779] Preparation Example P46: Polymer 522

[0780] Polymer 522 contains the following repeating units:

[0781] Following the procedure of Preparation Example P45, 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.

[0782] Preparation Example P47: Polymer 523

[0783] Polymer 523 contains the following repeating units:

[0784] Following the procedure of Preparation Example P45, 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.

[0785] Preparation Example P48: Polymer 524

[0786] Polymer 524 contains the following repeating units:

[0787] Following the procedure in Preparation Example P45, 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.

[0788] Preparation Example P49: Polymer 525

[0789] Polymer 525 contains the following repeating units:

[0790] Following the procedure of Preparation Example P45, 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.

[0791] Preparation Example P50: Polymer 526

[0792] Polymer 526 contains the following repeating units:

[0793] Following the procedure in Preparation Example P45, 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.

[0794] Preparation Example P51: Polymer 528

[0795] Polymer 528 contains the following repeating units:

[0796] Following the procedure in Preparation Example P45, 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.

[0797] Preparation Example P52: Polymer 545

[0798] Polymer 545 contains the following repeating units:

[0799] Following a similar process to that described in Preparation Example P1, A021 (100 mg, 1 eq), B021 (210 mg, 1.05 eq), and cesium fluoride (10.0 mg, 0.1 eq) were reacted to yield a white solid 545 (128 mg, yield 54.7%). Mw: 7.1 kDa.

[0800] Preparation Example P53: Polymer 547

[0801] Polymer 547 contains the following repeating units:

[0802] Following the procedure in Example P45, 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.

[0803] Preparation Example P54: Polymer 550

[0804] Polymer 550 contains the following repeating units:

[0805] Following the procedure in Example P45, 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.

[0806] Preparation Example P55: Polymer 609

[0807] Polymer 609 contains the following repeating units:

[0808] Following the procedure in Preparation Example P45, the starting materials were A004 (107 mg, 1 eq), B028 (245 mg, 1.05 eq), and cesium fluoride (9.7 mg, 0.1 eq), yielding a pale yellow solid 609 (80 mg, yield 30.1%). Mw: 17.9 kDa.

[0809] Preparation Example P56: Polymer 612

[0810] Polymer 612 contains the following repeating units:

[0811] Following the procedure of Preparation Example P45, 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.

[0812] Preparation Example P57: Polymer 614

[0813] Polymer 614 contains the following repeating units:

[0814] Following the procedure in Preparation Example P45, 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.

[0815] Preparation Example P58: Polymer 618

[0816] Polymer 618 contains the following repeating units:

[0817] Following the procedure in Preparation Example P45, 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.

[0818] Preparation Example P59: Polymer 619

[0819] Polymer 619 contains the following repeating units:

[0820] Following the procedure in Example P45, 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.

[0821] Preparation Example P60: Polymer E004

[0822] Polymer E004 contains the following repeating units:

[0823] 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.

[0824] Preparation Example P61: Polymer C004

[0825] Polymer C004 contains the following repeating units:

[0826] The raw materials M006 (525 mg, 1 eq) and M007 (152 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.

[0827] Preparation Example P62: Polymer R003

[0828] Polymer R003 contains the following repeating units:

[0829] 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.

[0830] Preparation Example P63: Polymer 723

[0831] Polymer 723 contains the following repeating units:

[0832] 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.

[0833] Preparation Example P64: Polymer 726

[0834] Polymer 726 contains the following repeating units:

[0835] 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). The reaction time was 2 h, yielding a pale yellow solid 726 (205 mg, yield 52.8%). Mw: 4.4 kDa.

[0836] Preparation Example P65: Polymer 727

[0837] Polymer 727 contains the following repeating units:

[0838] Following the procedure of Preparation Example P1, the starting materials were Al2O (251 mg, 1 eq), BO26 (301 mg, 1 eq), and cesium fluoride (13.1 mg, 0.1 eq), and the reaction time was 2 h, yielding a pale yellow solid 727 (207 mg, yield 47.9%). Mw: 6.2 kDa.

[0839] Preparation Example P66: Polymer 728

[0840] Polymer 728 contains the following repeating units:

[0841] 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.

[0842] Preparation Example P67: Polymer 732

[0843] Polymer 732 contains the following repeating units:

[0844] Following the procedure of Preparation Example P45, 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.

[0845] Preparation Example P68: Polymer 733

[0846] Polymer 733 contains the following repeating units:

[0847] Following the procedure in Preparation Example P45, 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.

[0848] Preparation Example P69: Polymer 734

[0849] Polymer 734 contains the following repeating units:

[0850] Following the procedure of Preparation Example P45, 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.

[0851] Preparation Example P70: Polymer 735

[0852] Polymer 735 contains the following repeating units:

[0853] Following the preparation process of Example P45, 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.

[0854] Preparation Example P71: Polymer 741

[0855] Polymer 741 contains the following repeating units:

[0856] Following the preparation process of Example P45, 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.

[0857] Preparation Example P72: Polymer 742

[0858] Polymer 742 contains the following repeating units:

[0859] Following the procedure of Preparation Example P45, 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.

[0860] Preparation Example P73: Polymer 744

[0861] Polymer 744 contains the following repeating units:

[0862] Following the procedure of Preparation Example P45, 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.

[0863] Preparation Example P74: Polymer 745

[0864] Polymer 745 contains the following repeating units:

[0865] Following the preparation process of Example P45, 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.

[0866] Preparation Example P75: Polymer 746

[0867] Polymer 746 contains the following repeating units:

[0868] Following the procedure of Preparation Example P45, 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.

[0869] Preparation Example P76: Polymer 750

[0870] Polymer 750 contains the following repeating units:

[0871] Following the procedure of Preparation Example P45, 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.

[0872] Preparation Example P77: Polymer 751

[0873] Polymer 751 contains the following repeating units:

[0874] Following the procedure of Preparation Example P45, 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.

[0875] Preparation Example P78: Polymer 752

[0876] Polymer 752 contains the following repeating units:

[0877] Following the procedure of Preparation Example P45, 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.

[0878] Preparation Example P79: Polymer 756

[0879] Polymer 756 contains the following repeating units:

[0880] Following the procedure of Preparation Example P45, 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.

[0881] Preparation Example P80: Polymer 757

[0882] Polymer 757 contains the following repeating units:

[0883] Following the procedure of Preparation Example P45, 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.

[0884] Preparation Example P81: Polymer 758

[0885] Polymer 758 contains the following repeating units:

[0886] Following the procedure of Preparation Example P45, 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.

[0887] Preparation Example P82: Polymer 759

[0888] Polymer 759 contains the following repeating units:

[0889] Following the procedure of Preparation Example P45, 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.

[0890] Preparation Example P83: Polymer 761

[0891] Polymer 761 contains the following repeating units:

[0892] 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.

[0893] Preparation Example P84: Polymer 763

[0894] Polymer 763 contains the following repeating units:

[0895] Following the procedure of Preparation Example P1, the starting materials were Al22 (238.8 mg, 1 eq), B026 (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.

[0896] Preparation Example P85: Polymer 764

[0897] Polymer 764 contains the following repeating units:

[0898] Following the procedure of Preparation Example P1, the starting materials were Al23 (264.6 mg, 1 eq), B026 (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.

[0899] Preparation Example P86: Polymer 765

[0900] Polymer 765 contains the following repeating units:

[0901] 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.

[0902] Preparation Example P87: Polymer 766

[0903] Polymer 766 contains the following repeating units:

[0904] 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), and the reaction time was 2 h, yielding a pale yellow solid 766 (135 mg, yield 41.5%). Mw: 11.9 kDa.

[0905] Preparation Example P88: Polymer 767

[0906] Polymer 767 contains the following repeating units:

[0907] 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.

[0908] Preparation Example P89: Polymer 768

[0909] Polymer 768 contains the following repeating units:

[0910] 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). The reaction time was 2 h, yielding a pale yellow solid 768 (161 mg, yield 47.6%). Mw: 5.1 kDa.

[0911] Preparation Example P90: Polymer 769

[0912] Polymer 769 contains the following repeating units:

[0913] 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.

[0914] Preparation Example P91: Polymer 772

[0915] Polymer 772 contains the following repeating units:

[0916] 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.

[0917] Preparation Example P92: Polymer 774

[0918] Polymer 774 contains the following repeating units:

[0919] 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.

[0920] Preparation Example P93: Polymer 779

[0921] Polymer 779 contains the following repeating units:

[0922] 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.

[0923] Preparation Example P94: Polymer 780

[0924] Polymer 780 contains the following repeating units:

[0925] Following the procedure described in Example P45, 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.

[0926] Preparation Example P95: Polymer 781

[0927] Polymer 781 contains the following repeating units:

[0928] Following the procedure described in Example P45, 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). The reaction time was 2 h, yielding a pale yellow solid 781 (252 mg, yield 63.3%). Mw: 10.1 kDa.

[0929] Preparation Example P96: Polymer 782

[0930] Polymer 782 contains the following repeating units:

[0931] Following the procedure of Preparation Example P1, 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), and the reaction time was 2 h, yielding a pale yellow solid 782 (198 mg, yield 54.7%). Mw: 5.1 kDa.

[0932] Preparation Example P97: Polymer 783

[0933] Polymer 783 contains the following repeating units:

[0934] Following the procedure of Preparation Example P1, the starting materials were Al32 (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.

[0935] Preparation Example P98: Polymer 784

[0936] Polymer 784 contains the following repeating units:

[0937] Following the procedure of Preparation Example P1, the starting materials were Al32 (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.

[0938] Preparation Example P99: Polymer 790

[0939] Polymer 790 contains the following repeating units:

[0940] 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.

[0941] Preparation Example P100: Polymer 791

[0942] Polymer 791 contains the following repeating units:

[0943] 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.

[0944] Example 3: Polymer molecular weight characterization

[0945] Polymers 244, 245, 246, 247, 250, 251, 262, 269, 276, 298, 301, 303, 307, 315, 328, 329, 334, 373, 376, 379, 380, 381, 382, ​​385, 388, 389, 392, 394, 395, 399, 431, 435, 474, 475, 521, 522, 523, 525, 526, 528, 545, 547, 550, 609, and 612 were prepared using 0.3M sodium acetate buffer (pH 5). Aqueous solutions (5 mg / mL) of the following: 614, 618, 619, 723, 726, 727, 728, 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, C004, and R003. Aqueous solutions containing DMSO (5% DMSO, 5 mg / mL) of polymers 273, 281, 282, 283, 288, 292, 293, 294, 299, 300, 326, 524, and E004 were prepared using 0.3 M sodium acetate buffer (pH 5). Samples were filtered through a 0.2 μm filter. Molecular weight was determined using aqueous size exclusion chromatography with differential detection 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), with an injector temperature of 4 °C, a column temperature of 30 °C, a flow rate of 0.6 mL / min, and a dn / dc ratio of 0.1659.

[0946] Example 4: Preparation of polymer-nucleic acid complex

[0947] This embodiment provides the following exemplary method for preparing the polymer-nucleic acid complex.

[0948] 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 a 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, 380, 381, 382, ​​385, 388, 389, 392, 394, 395, 399, 431, 435, 474, 475, 521, 522, 523, 525, 526, 528, 545, 547, 550, 609, 612, 614, 618, 619, 723, 726 Complexes of 727, 728, 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, 380, 522, 523, 525, 526, 528, 547, 550, and 791 with the pCMV-luc plasmid were prepared using method 1.

[0949] 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 incubated 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.

[0950] Example 5: Performance Characterization of Polymer-Nucleic Acid Complexes

[0951] A. Measurement of average particle size, particle dispersion index (PDI), and zeta surface potential of polymer-nucleic acid complex nanoparticles

[0952] 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.

[0953] B. Measurement of polymer-nucleic acid complex encapsulation efficiency

[0954] 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 the unencapsulated Luc-mRNA or pCMV-luc sample. No bright bands were observed after normalization, indicating 100% encapsulation of the complex. Results are shown in Table 4 or Table 5.

[0955] Table 4. Characterization results of the polymer / Luc-mRNA complex

[0956] Table 5. Characterization results of the polymer / pCMV-luc plasmid complex

[0957] Example 6: Cell safety and transfection performance of the complex

[0958] 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).

[0959] 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.

[0960] 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 light set to 560 nm and emission light set to 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.

[0961] 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). This demonstrates that the polymer-nucleic acid complex described herein effectively transfects cells and successfully expresses the loaded nucleic acid, with a transfection efficiency significantly higher than the negative control and all comparative examples.

[0962] 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”.

[0963] 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.

[0964] 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, A comprises at least one nitrogen-containing group; 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 a2 are each independently an integer from 1 to 16; b1 and b2 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 comprises at least one protonatable nitrogen-containing group. The polycarbonate or salt thereof according to claim 1 or 2, wherein The at least one nitrogen-containing group is located in the main chain of the polycarbonate; in particular, at least one nitrogen atom of the at least one nitrogen-containing group is located in the main chain of the polycarbonate; more particularly, the at least one protonatable nitrogen-containing group is located in the main chain of the polycarbonate; more particularly, at least one nitrogen atom of 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 2, 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 combinations thereof; in particular, the amino group is selected from the group consisting of a secondary amino group, a tertiary amino group, a quaternary amino group, and combinations thereof; in particular, 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 ring of a 3- to 10-membered saturated or unsaturated aliphatic or aromatic heterocyclic group having at least one ring nitrogen atom, and combinations thereof. The polycarbonate or salt thereof according to claim 1, wherein, The polycarbonate further comprises one or more additional groups selected from the group consisting of a redox-sensitive group, a hydrocarbon group, an oxygen-containing group, and combinations thereof. A method of making polycarbonate comprising polymerizing at least one monomer having the formula II with at least one monomer having the formula III: in formula II and formula III, A 1 and A 2 each independently contains one or more groups independently of one another having the formula IV: wherein, M is selected from the group consisting of a nitrogen-containing group, a redox-sensitive group, a hydrocarbylene group, an oxygen-containing group, and combinations thereof; x1 and x2 are each independently an integer from 0 to 2; y1 and y2 are each independently an integer from 0 to 16; 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; wherein A 1 and A 2 has at least one nitrogen-containing group; wherein, for A 1 and A 2 when comprising two or more groups independently of Formula IV, the two or more groups independently of Formula IV are connected to each other via L M wherein L M is a bond, a carbonate linkage, or a C 1-16 alkylene group; and wherein the 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 combinations 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; in particular, the composition is a pharmaceutical composition. A method of delivering or expressing at least one nucleic acid in a cell or a subject, the method comprising: - providing a polycarbonate nucleic acid complex according to claim 7, the complex being formed by complexing the at least one nucleic acid with the polycarbonate or salt thereof; and - contacting the cell or the subject with the complex. Use of the polycarbonate or salt thereof according to claim 1 as a nucleic acid carrier for delivering the nucleic acid to a cell or a subject.

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