Peg lipid and use thereof

Abstract

The present disclosure relates to a PEG lipid and the use thereof. The PEG lipid can be used for preparing a lipid nanoparticle that delivers an active ingredient. The present disclosure further relates to a pharmaceutical composition and a lipid nanoparticle comprising the PEG lipid, as well as the use thereof in preparing a pharmaceutical.

Description

[Corresponding to Rule 26, amended on November 25, 2025] PEG lipid and its application [Corresponding to Rule 26, amended on November 25, 2025] TECHNICAL FIELD

[0001] [Corresponding to Rule 26, amended on November 25, 2025] The present invention belongs to the field of biotechnology, and relates to a PEG lipid and its application. [Corresponding to Rule 26, amended on November 25, 2025] BACKGROUND

[0002] [Corresponding to Rule 26, amended on November 25, 2025] Nucleic acid drugs exert their effects on RNA or DNA, including mRNA, antisense oligonucleotides (ASOs), microRNAs (miRNAs), small interfering RNAs (siRNAs), and aptamers, etc. Studies have found that nucleic acid drugs have great therapeutic potential in the fields of cancer treatment, infectious disease prevention, and genetic disease treatment, etc.

[0003] [Corresponding to Rule 26, amended on November 25, 2025] To avoid being degraded by nucleases in the blood, nucleic acid drugs need to be delivered by a delivery carrier to the body to exert their efficacy. Common delivery carriers for nucleic acid drugs include lipid nanoparticles (LNPs). Lipid nanoparticles generally contain cationic lipids, phospholipids, helper lipids, and PEG lipids, wherein the PEG lipid can both drive self-assembly and prevent particle aggregation, making the preparation of lipid nanoparticles easier, and it is generally believed that the PEG lipid can prolong the stability and residence time of the lipid nanoparticles in the blood circulation. However, recent studies have found that repeated injection of lipid nanoparticles containing PEG lipids can cause an immune response, which leads to a decrease in the in vivo circulation time of the lipid nanoparticles and an increase in their accumulation in the liver and spleen, a phenomenon known as "accelerated blood clearance" (ABC) phenomenon. The accelerated blood clearance phenomenon of PEG lipids poses a serious challenge to their development and clinical application. [Corresponding to Rule 26, amended on November 25, 2025] SUMMARY

[0004] [Corresponding to Rule 26, amended on November 25, 2025] In a first aspect, the present disclosure provides a PEG lipid as shown in formula (I):

[0005] [Corresponding to Rule 26, amended on November 25, 2025] or a salt thereof, or a stereoisomer thereof, wherein

[0006] [Corrected according to Rule 26 25.11.2025] R1is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0007] [Corrected according to Rule 26 25.11.2025] Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond,

[0008] [Corrected according to Rule 26 25.11.2025] Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond,

[0009] [Corrected according to Rule 26 25.11.2025] Q1is -C(=O)-, -OC(=O)-, or a bond,

[0010] [Corrected according to Rule 26 25.11.2025] Q2is -C(=O)-, or a bond,

[0011] [Corrected according to Rule 26 25.11.2025] Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond,

[0012] [Corrected according to Rule 26 25.11.2025] L1is C1-C 20 alkylene or a bond,

[0013] [Corrected according to Rule 26 25.11.2025] A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond,

[0014] [Corrected according to Rule 26 25.11.2025] R2is C4-C 24 alkyl,

[0015] [Corrected according to Rule 26 25.11.2025] The total number of carbon atoms of L1and R2is 10-35,

[0016] [Corrected according to Rule 26 25.11.2025] L2is C1-C4alkylene,

[0017] [Corrected according to Rule 26 25.11.2025] L3is C1-C4alkylene,

[0018] [Corrected according to Rule 26 25.11.2025] R3is H or C1-C3alkyl,

[0019] [Corrected according to Rule 26 25.11.2025] n is any integer from 30 to 60.

[0020] [Corrected according to Rule 26 25.11.2025] In a second aspect, the present disclosure also provides a PEG lipid as shown in Formula (IX):

[0021] [Corrected according to Rule 26 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein

[0022] [Corrected according to Rule 26 25.11.2025] R1is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0023] [Corrected according to Rule 26 25.11.2025] Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond,

[0024] [Corrected according to Rule 26 25.11.2025] Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond,

[0025] [Corrected according to Rule 26 25.11.2025] Q1is -C(=O)-, -OC(=O)-, or a bond,

[0026] [Corrected according to Rule 26 25.11.2025] Q2is -C(=O)-, or a bond,

[0027] [Corrected according to Rule 26 25.11.2025] Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond,

[0028] [Corrected according to Rule 26 25.11.2025] L1is C1-C 20 alkylene or a bond,

[0029] [Corrected according to Rule 26 25.11.2025] A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond,

[0030] [Corrected according to Rule 26 25.11.2025] R2is C4-C 24 alkyl,

[0031] [Corrected according to Rule 26 25.11.2025] The total number of carbon atoms of L1and R2is 10-35,

[0032] [Corrected according to Rule 26 25.11.2025] L2is C1-C4alkylene,

[0033] [Amended according to Rule 26 on 25.11.2025] L3 is C1-C4 alkylene,

[0034] [Amended according to Rule 26 on 25.11.2025] R3 is H or C1-C3 alkyl,

[0035] [Amended according to Rule 26 on 25.11.2025] n is any integer from 30 to 60,

[0036] [Amended according to Rule 26 on 25.11.2025] L1 forms a four-, five- or six-membered ring with A1 and R2.

[0037] [Amended according to Rule 26 on 25.11.2025] In some embodiments, L1 in formula (IX) is C1-C 20 alkylene.

[0038] [Amended according to Rule 26 on 25.11.2025] In a third aspect, the present disclosure also provides a PEG lipid as shown in formula (X):

[0039] [Amended according to Rule 26 on 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein

[0040] [Amended according to Rule 26 on 25.11.2025] R1 is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0041] [Amended according to Rule 26 on 25.11.2025] Z1 and Z2 are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2- or a bond,

[0042] [Amended according to Rule 26 on 25.11.2025] Q1 is -C(=O)-, -OC(=O)- or a bond,

[0043] [Amended according to Rule 26 on 25.11.2025] Q2 is -C(=O)- or a bond,

[0044] [Amended according to Rule 26 on 25.11.2025] Q3 is -C(=O)O-, -OC(=O)-, -O-, -S- or a bond,

[0045] [Amended according to Rule 26 on 25.11.2025] L1 is C1-C 20 alkylene or a bond,

[0046] [Corrected according to Rule 26 25.11.2025] A1is -C(=0)0-, -OC(=0)-, -OC(=0)0-, -0-, -S-, or a bond,

[0047] [Corrected according to Rule 26 25.11.2025] R2is C4-C 24 hydrocarbyl,

[0048] [Corrected according to Rule 26 25.11.2025] The total number of carbon atoms in L1and R2is 10-35,

[0049] [Corrected according to Rule 26 25.11.2025] L2is C1-C4 hydrocarbylene,

[0050] [Corrected according to Rule 26 25.11.2025] L3is C1-C4 hydrocarbylene,

[0051] [Corrected according to Rule 26 25.11.2025] R3is H or C1-C3 hydrocarbyl,

[0052] [Corrected according to Rule 26 25.11.2025] n is any integer from 30-60,

[0053] [Corrected according to Rule 26 25.11.2025] Z2forms a four-, five-, or six-membered ring with Q3, L1, A1, and R2.

[0054] [Corrected according to Rule 26 25.11.2025] In some embodiments, in formula (X), Z1is -L2-CH(OR3)-L3-, -L2-C(=0)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond, and Z2is -L2-CH(OR3)-L3-, -L2-C(=0)-L3-, -L2-CH(R3)-L3-, or -L2-.

[0055] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H,

[0056] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H,

[0057] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid has the structure of formula (II):

[0058] [Corrected according to Rule 26 25.11.2025] wherein R1is not H and is not unsubstituted methyl.

[0059] [Amended according to Rule 26 25.11.2025] In some embodiments, R1is H-Z1-Q1-, A1is a bond, and Z1and Q1are not simultaneously a bond; preferably, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-. In some such embodiments, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-, and Q1is -C(=O)-, -OC(=O)-, or a bond. In preferred embodiments, L2is C1-C3alkylene, L3is C1-C3alkylene, and R3is H or C1-C3alkyl. In more preferred embodiments, R1is

[0060] [Amended according to Rule 26 25.11.2025] In some embodiments, R1is H-Z1-Q1-, wherein Z1is not a bond and is not unsubstituted methylene when Q1is a bond. In preferred embodiments, R1is H-Z1-Q1-, Z1is -L2-, Q1is -C(=O)-, and A1is a bond. In such embodiments, L2is C1-C3alkylene. In preferred embodiments, R1is

[0061] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid is of the formula (II-1):

[0062] [Amended according to Rule 26 25.11.2025] wherein A1is -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0063] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid is of the formula (II-1-1):

[0064] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid has a structure of formula (III), formula (IV), formula (V), formula (VI), or formula (VII):

[0065] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid has a structure of:

[0066] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms of L1and R2is 12-25.

[0067] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms of L1and R2is 16-22.

[0068] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is C1-C8 hydrocarbylene or a bond.

[0069] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is C1-C8 alkylene, C2-C8 alkenylene, or C2-C8 alkynylene.

[0070] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is C7-C 22 hydrocarbyl.

[0071] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is C7-C 22 alkyl, C7-C 22 alkenyl, or C7-C 22 alkynyl.

[0072] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2is C1-C3 hydrocarbylene.

[0073] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2is C1-C3 alkylene, C2-C3 alkenylene, or C2-C3 alkynylene.

[0074] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3is C1-C2 hydrocarbylene.

[0075] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3is C1-C2 alkylene, C2 alkenylene, or C2 alkynylene.

[0076] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3is H or C1-C3 alkyl.

[0077] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 30-55.

[0078] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 35 to 50.

[0079] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 40 to 50.

[0080] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid is:

[0081] [Corrected according to Rule 26 25.11.2025] In another aspect, the present disclosure provides use of the PEG lipid of any of the above embodiments in the manufacture of a lipid nanoparticle.

[0082] [Corrected according to Rule 26 25.11.2025] In another aspect, the present disclosure provides a lipid nanoparticle comprising the PEG lipid of any of the above embodiments.

[0083] [Corrected according to Rule 26 25.11.2025] In some embodiments, the lipid nanoparticle further comprises an active ingredient. In some embodiments, the active ingredient is a pharmaceutically active ingredient. In some embodiments, the active ingredient is a nucleic acid. In some embodiments, the pharmaceutically active ingredient is a nucleic acid.

[0084] [Corrected according to Rule 26 25.11.2025] In some embodiments, the lipid nanoparticle further comprises one or more of a cationic lipid, a helper lipid, a structural lipid, and a polymeric lipid.

[0085] [Corrected according to Rule 26 25.11.2025] In another aspect, the present disclosure provides a composition comprising the lipid nanoparticle of any of the above embodiments, and a biologically acceptable carrier.

[0086] [Corrected according to Rule 26 25.11.2025] In another aspect, the present disclosure provides a pharmaceutical composition comprising the lipid nanoparticle of any of the above embodiments, and a pharmaceutically acceptable carrier.

[0087] [Corrected according to Rule 26 25.11.2025] In another aspect, the present disclosure provides use of the lipid nanoparticle of any of the above embodiments or the pharmaceutical composition of any of the above embodiments in the manufacture of a medicament.

[0088] [Corrected according to Rule 26 25.11.2025] In some embodiments, the medicament is for gene therapy, gene vaccination, protein replacement therapy, antisense therapy or treatment by interfering RNA.

[0089] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-mentioned lipid particle is used as a carrier for transferring or delivering an active ingredient. In some embodiments, the active ingredient is a pharmaceutically active ingredient. In some embodiments, the above-mentioned pharmaceutically active ingredient is a nucleic acid.

[0090] [Corrected according to Rule 26 25.11.2025] In some embodiments, the medicament is for the treatment and / or prevention of a disease.

[0091] [Corrected according to Rule 26 25.11.2025] In some embodiments, the medicament is for the treatment and / or prevention of one or more of the following: rare diseases, cancer, infectious diseases, autoimmune diseases, metabolic diseases, neurological diseases, cardiovascular diseases, transplant rejection, inflammatory reactions, genetic diseases and musculoskeletal diseases.

[0092] [Corrected according to Rule 26 25.11.2025] In some embodiments, the rare disease comprises one or more of the following: osteogenesis imperfecta, Wilson's disease, spinal muscular atrophy, Huntington's disease, Rett syndrome, amyotrophic lateral sclerosis, Duchenne muscular dystrophy, Friedrich's ataxia, methylmalonic acidemia, cystic fibrosis, glycogen storage disease la, glycogen storage disease III, Crigler-Najjar syndrome, ornithine transcarbamylase deficiency, propionic acidemia, phenylketonuria, hemophilia A, hemophilia B, beta-thalassemia, Lafora disease, Dravet syndrome, Alexander disease, Leber congenital amaurosis, myelodysplastic syndrome and CBS-deficient homocystinuria.

[0093] [Corrected according to Rule 26 25.11.2025] In some embodiments, the cancer comprises one or more of the following: hematological malignancies, lung cancer, liver cancer, kidney cancer, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, pancreatic cancer, brain cancer, prostate cancer, gallbladder cancer, ovarian cancer, breast cancer, cervical cancer, endometrial cancer, bladder cancer and melanoma.

[0094] [Corrected according to Rule 26 25.11.2025] In some embodiments, the infectious disease comprises a disease caused by one or more of a viral, fungal and bacterial infection.

[0095] [Corrected according to Rule 26 25.11.2025] In some embodiments, the autoimmune disease comprises one or more of the following: acute idiopathic thrombocytopenic purpura, chronic idiopathic thrombocytopenic purpura, systemic lupus erythematosus, rheumatoid arthritis, psoriasis, inflammatory bowel disease, multiple sclerosis, celiac disease, type 1 diabetes, and diffuse toxic goiter.

[0096] [Corrected according to Rule 26 25.11.2025] In some embodiments, the genetic disease comprises one or more of the following: hemophilia, thalassemia, and Gaucher's disease.

[0097] [Corrected according to Rule 26 25.11.2025] In some embodiments, the neurological disease comprises one or more of the following: amyotrophic lateral sclerosis, Alzheimer's disease, and glioma.

[0098] [Corrected according to Rule 26 25.11.2025] In some embodiments, the drug is a vaccine.

[0099] [Corrected according to Rule 26 25.11.2025] In some embodiments, the drug is a nucleic acid drug, wherein the nucleic acid comprises at least one of the following: RNA, messenger RNA (mRNA), antisense oligonucleotide, DNA, plasmid, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA), and Cas9 mRNA.

[0100] [Corrected according to Rule 26 25.11.2025] In yet another aspect, the present disclosure provides use of the lipid nanoparticle described above in the manufacture of a carrier for the transfer or delivery of an active ingredient. In some embodiments, the active ingredient is a pharmaceutically active ingredient, such as a nucleic acid. In other embodiments, the active ingredient is a transfection reagent or a detection reagent.

[0101] [Corrected according to Rule 26 25.11.2025] In some embodiments, the nucleic acid comprises at least one of the following: RNA, messenger RNA (mRNA), antisense oligonucleotide, DNA, plasmid, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA), and Cas9 mRNA. [Corrected according to Rule 26 25.11.2025] BRIEF DESCRIPTION OF DRAWINGS

[0102] [Rule 26 correction 25.11.2025] Figure 1 shows the results of detection of hEPO concentration in serum of rats after multiple dosing. "001" in Figure 1 represents a lipid nanoparticle comprising the PEG lipid numbered 001 in Table 1, and so on.

[0103] [Rule 26 correction 25.11.2025] Figure 2 shows the results of detection of anti-PEG IgG antibody in serum of rats after multiple dosing. "001" in Figure 2 represents a lipid nanoparticle comprising the PEG lipid numbered 001 in Table 1, and so on.

[0104] [Rule 26 correction 25.11.2025] Figure 3 shows the results of detection of cytokine IL-6 in serum of rats after dosing. "001" in Figure 3 represents a lipid nanoparticle comprising the PEG lipid numbered 001 in Table 1, and so on.

[0105] [Rule 26 correction 25.11.2025] Figure 4 shows the results of detection of cytokine IFN-γ in serum of rats after dosing. "001" in Figure 4 represents a lipid nanoparticle comprising the PEG lipid numbered 001 in Table 1, and so on.

[0106] [Rule 26 correction 25.11.2025] DETAILED DESCRIPTION

[0107] [Rule 26 correction 25.11.2025] I. DEFINITIONS

[0108] [Rule 26 correction 25.11.2025] All patents, patent applications, scientific publications, manufacturer's specifications and manuals, and the like, cited herein are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such disclosure.

[0109] [Rule 26 correction 25.11.2025] Unless defined otherwise, all scientific and technical terms used herein have the meanings that are commonly understood by one of ordinary skill in the art to which this disclosure belongs. Also, all patents, patent applications, scientific publications, manufacturer's specifications and manuals, and the like, cited herein are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such disclosure.

[0110] [Amended according to Rule 26, 25.11.2025] As used herein, the expressions “comprising,” “including,” “containing,” and “having” are open-ended, meaning that they include the listed elements, steps, or components but do not exclude other unlisted elements, steps, or components. The expression “consisting of” excludes any unspecified elements, steps, or components. The expression “substantially consisting of” means that the scope is limited to the specified elements, steps, or components, plus optional elements, steps, or components that do not significantly affect the essential and novel nature of the claimed subject matter. It should be understood that the expressions “substantially consisting of” and “consisting of” are encompassed within the meaning of the expression “including.”

[0111] [Amended according to Rule 26, 25.11.2025] As used herein, unless the context otherwise indicates, the singular forms of “a,” “an,” and “the,” and similar references used in the context of describing the invention (particularly in the context of the claims) shall be interpreted to cover both the singular and plural. The terms “one or more” or “at least one” cover 1, 2, 3, 4, 5, 6, 7, 8, 9, or more. The terms “at least one” or “one or more” cover 1, 2, 3, 4, 5, 6, 7, 8, 9, or more.

[0112] [Amended according to Rule 26, 25.11.2025] The numerical ranges described herein should be understood to encompass any and all subranges contained herein. For example, the range “1 to 10” should be understood to include not only the explicitly stated values ​​of 1 and 10, but also any individual value (e.g., 2, 3, 4, 5, 6, 7, 8, and 9) and subranges (e.g., 1 to 2, 1.5 to 2.5, 1 to 3, 1.5 to 3.5, 2.5 to 4, 3 to 4.5, etc.) within the range of 1 to 10. As another example, “C1 to C6 alkyl” is intended to encompass C1, C2, C3, C4, C5, C6, C… 1～6 C 1～5 C 1～4 C 1～3 C 1～2 C 2～6 C 2～5 C 2～4 C 2～3 C 3～6 C 3～5 C 3～4 C 4～6 C 4～5 and C 5～6 Alkyl groups. This principle also applies to ranges where only one value is used as the minimum or maximum value.

[0113] [Amended 25.11.2025 in accordance with Rule 26] As used herein, the terms “and / or,” “any combination thereof,” and their grammatical equivalents are used interchangeably. These terms may express, specifically, any combination. For example, the phrases “A, B, and / or C” or “A, B, C, or any combination thereof” may refer to “A alone; B alone; C alone; A and B; B and C; A and C; and A, B, and C.”

[0114] [Amended according to Rule 26 25.11.2025] Unless otherwise stated, all methods described herein may be performed in any suitable order.

[0115] [Amended according to Rule 26, 25.11.2025] As used herein, the term "hydrocarbon group" means the group remaining after an aliphatic hydrocarbon loses one hydrogen atom, including straight-chain or branched, saturated or unsaturated hydrocarbon groups. Hydrocarbon groups include, but are not limited to, alkyl, alkenyl, and alkynyl groups. In some embodiments, the hydrocarbon group has 1 to 24 carbon atoms (C1-C2). 24 Hydrocarbon group), for example, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms (C1, C2, C3, ... C2). 21 C 22 C 23 Or C 24 Hydrocarbon groups). Examples of hydrocarbon groups include, but are not limited to, C1-C1 groups. 24 Hydrocarbon group, C1-C 22 Hydrocarbon group, C1-C 20 Hydrocarbon group, C1-C 18 Hydrocarbon group, C1-C 16 Hydrocarbon group, C1-C 12 Hydrocarbon group, C1-C 10 Hydrocarbon group, C1-C8 hydrocarbon group, C1-C7 hydrocarbon group, C1-C6 hydrocarbon group, C1-C4 hydrocarbon group, C1-C3 hydrocarbon group, C1-C2 hydrocarbon group, C2-C8 hydrocarbon group, C2-C4 hydrocarbon group, C4-C8 hydrocarbon group, C4-C9 hydrocarbon group, C5-C8 hydrocarbon group, C3 hydrocarbon group, C4 hydrocarbon group, C5 hydrocarbon group, C6 hydrocarbon group, C7 hydrocarbon group, C8 hydrocarbon group, C9 hydrocarbon group, C 10 hydrocarbon group, C 11 hydrocarbon group, C 12 hydrocarbon group, C 13 hydrocarbon group, C 14 hydrocarbon group, C 15 hydrocarbon group, C 16 hydrocarbon group, C 17 hydrocarbon group, C 18 hydrocarbon group, C 19 hydrocarbon group, C 20 hydrocarbon group, C 21 hydrocarbon group and C22 Hydrocarbyl. Unless otherwise explicitly indicated otherwise in the present specification, a hydrocarbyl group is optionally substituted, the substituents being in accordance with the definition of "optionally substituted" below. In certain embodiments, a hydrocarbyl group has zero branches (i.e. is linear), one branch, two branches, or multiple branches.

[0116] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkylene" means a divalent radical resulting from the removal of one hydrogen atom from each of two aforementioned alkyl groups. Unless otherwise explicitly indicated otherwise in the present specification, an alkylene group is also optionally substituted.

[0117] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkyl" is a straight-chain or branched saturated monovalent hydrocarbon group. In some embodiments, an alkyl group has 1 to 24 carbon atoms (Ci-C 24 alkyl), for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 carbon atoms (Ci, C2, C3,... C 21 , C 22 , C 23 , or C 24 alkyl). Examples of alkyl groups include, but are not limited to, Ci-C 24 alkyl, Ci-C 22 alkyl, Ci-C 20 alkyl, Ci-C 18 alkyl, Ci-C 16 alkyl, Ci-C 12 alkyl, Ci-C 10 alkyl, Ci-C8alkyl, Ci-C7alkyl, Ci-C6alkyl, Ci-C4alkyl, Ci-C3alkyl, Ci-C2alkyl, C2-C8alkyl, C2-C4alkyl, C4-C8alkyl, C4-C9alkyl, C5-C8alkyl, methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, i-butyl, t-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, and tridec-7-yl. Unless otherwise explicitly indicated otherwise in the present specification, an alkyl group is optionally substituted.

[0118] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkylene" means a divalent radical resulting from the removal of one hydrogen atom from each of two aforementioned alkyl groups. Unless otherwise explicitly indicated otherwise in the present specification, an alkylene group is also optionally substituted.

[0119] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkenyl" is a straight-chain or branched monovalent hydrocarbon group, which contains one or more double bonds (C=C). In some embodiments, the alkenyl group has 2 to 24 carbon atoms (C2-C 24 alkenyl), for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 carbon atoms (C2, C3, C4,... C 21 , C 22 , C 23 , or C 24 alkenyl), and has 1, 2, 3, 4, or more double bonds. Alkenyl groups include, but are not limited to, C2-C 24 alkenyl, C2-C 22 alkenyl, C2-C 20 alkenyl, C2-C 18 alkenyl, C2-C 16 alkenyl, C2-C 12 alkenyl, C2-C 10 alkenyl, C2-C8alkenyl, C2-C7alkenyl, C2-C6alkenyl, C2-C4alkenyl, C2-C3alkenyl, C4-C8alkenyl, C4-C9alkenyl, C5-C8alkenyl, which has 1, 2, 3, 4, or more double bonds. Some more specific examples include, but are not limited to, ethenyl, propenyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-2-enyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hept-1-enyl, hept-2-enyl, hept-3-enyl, oct-1-enyl, oct-2-enyl, oct-3-enyl, non-1-enyl, non-2-enyl, and non-3-enyl. In some embodiments, the alkenyl group has 1 double bond. Unless specifically indicated otherwise in the specification, the alkenyl group is optionally substituted.

[0120] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkenylene" means the divalent radical which remains after the loss of 1 hydrogen atom from an alkenyl group as described above. Unless specifically indicated otherwise in the specification, the alkenylene group is also optionally substituted.

[0121] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkynyl" is a straight-chain or branched monovalent hydrocarbon group, which contains one or more triple bonds (C≡C). In some embodiments, the alkynyl group has 2 to 24 carbon atoms (C2-C 24 alkynyl), for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 carbon atoms (C2, C3, C4,... C21 , C 22 , C 23 or C 24 alkynyl), and having 1, 2, 3, 4, or more triple bonds. Alkynyl includes, but is not limited to, C2-C8alkynyl, C2-C7alkynyl, C2-C6alkynyl, C2-C4alkynyl, C2-C3alkynyl, C4-C8alkynyl, C4-C9alkynyl, C5-C8alkynyl, which have 1, 2, 3, 4, or more triple bonds. Some more specific examples include, but are not limited to, ethynyl, propynyl, but-1-ynyl, but-2-ynyl, pent-1-ynyl, pent-2-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hept-1-ynyl, hept-2-ynyl, hept-3-ynyl, oct-1-ynyl, oct-2-ynyl, oct-3-ynyl, non-1-ynyl, non-2-ynyl, and non-3-ynyl. In some embodiments, the alkynyl group has 1 triple bond. Unless specifically indicated otherwise in the specification, alkynyl groups are optionally substituted. 24 alkynyl, C2-C 22 alkynyl, C2-C 20 alkynyl, C2-C 18 alkynyl, C2-C 16 alkynyl, C2-C 12 alkynyl, C2-C 10 alkynyl, C2-C8alkynyl, C2-C7alkynyl, C2-C6alkynyl, C2-C4alkynyl, C2-C3alkynyl, C4-C8alkynyl, C4-C9alkynyl, C5-C8alkynyl, which have 1, 2, 3, 4, or more triple bonds. Some more specific examples include, but are not limited to, ethynyl, propynyl, but-1-ynyl, but-2-ynyl, pent-1-ynyl, pent-2-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hept-1-ynyl, hept-2-ynyl, hept-3-ynyl, oct-1-ynyl, oct-2-ynyl, oct-3-ynyl, non-1-ynyl, non-2-ynyl, and non-3-ynyl. In some embodiments, the alkynyl group has 1 triple bond. Unless specifically indicated otherwise in the specification, alkynyl groups are optionally substituted.

[0122] [Corrected according to Rule 26 25.11.2025] As used herein, the term "alkynylene" means a divalent radical remaining after further loss of 1 hydrogen atom from an alkynyl group, as described above. Unless specifically indicated otherwise in the specification, alkynylene groups are also optionally substituted.

[0123] [Corrected according to Rule 26 25.11.2025] The term "optionally substituted" as used herein means that 1 or more hydrogen atoms attached to an atom or group are independently either unsubstituted, or substituted with 1 or more (e.g., 1, 2, 3, or 4) substituents. The substituents can be independently selected from, but are not limited to: halogen (e.g., chlorine, bromine, fluorine, or iodine), carboxylic acid (e.g., -C(=0)OH), oxygen (e.g., =0), sulfur (e.g., =S), hydroxyl (e.g., -OH), ester (e.g., -C(=0)ORiii or -OC(=0)Riii), aldehyde (e.g., -C(=0)H), carbonyl (e.g., -C(=0)Riii, or represented by C=0), acyl halide (e.g., -C(=0)X, where X is selected from bromine, fluorine, chlorine, or iodine), carbonate (e.g., -OC(=0)ORiii), alkoxy (e.g., -ORiii), acetal (e.g., -C(ORiii)2Riii, where each ORiii is the same or different alkoxy group), phosphate (e.g., P(=0)4 3-), thiol (e.g., -SH), sulfoxide (e.g., -S(=0)Riii), sulfmic acid (e.g., -S(=0)OH), sulfonic acid (e.g., -S(=0)2OH), thial (e.g., -C(=S)H), sulfate (e.g., S(=0)4 2-sulfonyl (e.g., -S(=0)2Riii), sulfinyl (e.g., -S(=0)Riii), amide (e.g., -C(=0)N(Riii)2or -N(Riii)C(=0)Riii), azido (e.g., -N3), nitro (e.g., -NO2), cyano (e.g., -CN), isocyano (e.g., -NC), acyloxy (e.g., -OC(=0)Riii), amino (e.g., -N(Riii)2, -N(Riii)H, or -NH2), carbamoyl (e.g., -OC(=0)N(Riii)2, -OC(=0)N(Riii)H, or -OC(=0)NH2), sulfamide (e.g., -S(=0)2N(Riii)2, -S(=0)2N(Riii)H, -S(=0)2NH2, -N(Riii)S(=0)2Riii, -N(H)S(=0)2Riii, -N(Riii)S(=0)2H, or -N(H)S(=0)2H), alkyl, alkenyl, alkynyl, cycloalkyl (e.g., cycloalkyl, cycloalkenyl, or cycloalkynyl), heterocycloalkyl (e.g., heterocycloalkyl containing one or more heteroatoms selected from S, N, and O, or heterocycloalkenyl containing one or more heteroatoms selected from S, N, and O), aryl (e.g., phenyl or fused ring aryl), heteroaryl (e.g., 8- to 10-membered bicyclic heteroaryl containing 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur), -C(=0)SRiii, -C(=N-CN)N(Riii)2, -C(=N-O-CH3)N(Riii)2, -C(=N-SO2-NH2)N(Riii)2, -C(=CH-NO2)N(Riii)2, -OC(=0)N(Riii)2, -CHN(Riii)N(Riii)2, -C(=0)N(Riii)ORiii, -N(Riii)2C(=0)ORiii, -OP(=0)(ORiii)2, -P(=0)(ORiii)2, -N(ORiii)C(=0)Riii, -N(ORiii)S(=0)2Riii, -N(ORiii)C(=0)ORiii, -N(ORiii)C(=0)N(Riii)2, -N(ORiii)C(=S)N(Riii)2, -N(ORiii)C(NRiii)N(Riii)2, -N(ORiii)C(CHRiii)N(Riii)2. In any of the foregoing, Riii is hydrogen, or alkyl, or alkenyl, or alkynyl, or heteroalkyl, or heteroalkenyl, or heteroalkynyl, as defined herein. In some embodiments, Riii is hydrogen, or C1-C6alkyl, or C2-C6alkenyl, or C2-C6alkynyl, or C2-C6heteroalkyl, or C3-C6heteroalkenyl, or C3-C6heteroalkynyl, as defined herein. 12 In some embodiments, R1is hydrogen, or C1-C6alkyl, or C2-C6alkenyl, or C2-C6alkynyl, or C2-C6heteroalkyl, or C3-C6heteroalkenyl, or C3-C6heteroalkynyl, as defined herein. 12 In some embodiments, R1is hydrogen, or C1-C6alkyl, or C2-C6alkenyl, or C2-C6alkynyl, or C2-C6heteroalkyl, or C3-C6heteroalkenyl, or C3-C6heteroalkynyl, as defined herein. 12 In some embodiments, R1is hydrogen, or C1-C6alkyl, or C2-C6alkenyl, or C2-C6alkynyl, or C2-C6heteroalkyl, or C3-C6heteroalkenyl, or C3-C6heteroalkynyl, as defined herein. 12 In some embodiments, R1is hydrogen, or C1-C6alkyl, or C2-C6alkenyl, or C2-C6alkynyl, or C2-C6heteroalkyl, or C3-C6heteroalkenyl, or C3-C6heteroalkynyl, as defined herein.12 Heterene group, or C3-C 12 The substituent itself may be further substituted with one or more substituents as defined herein. For example, the C1-C6 alkyl group that is a substituent may be further substituted with one or more substituents as described herein.

[0124] [Amended 25.11.2025 in accordance with Rule 26] As used herein, the term “halogenated” or “halogen” group is defined as including F, Cl, Br or I.

[0125] [Amended according to Rule 26, 25.11.2025] As used herein, “pharmaceuticalally acceptable salt” means an acid addition salt or base addition salt of the compounds disclosed herein, which retains the biological efficacy and properties of the compounds disclosed herein and is generally not a biologically or otherwise undesirable salt. In many cases, the compounds of the present invention are capable of forming acid salts and / or base salts due to the presence of amino and / or carboxyl groups or similar groups.

[0126] [Amended according to Rule 26, 25.11.2025] Pharmaceutically acceptable acid addition salts can be formed with inorganic and / or organic acids from compounds of this disclosure, said inorganic acids being, for example, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid; said organic acids being, for example, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, camphoric acid, camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, Formic acid, fumaric acid, galactosic acid, gentic acid, glucoheponic acid, gluconic acid, glucuronic acid, glutamic acid, glutamate, 2-oxoglutamate, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucoic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanate, p-toluenesulfonic acid, trifluoroacetic acid, and undecenoic acid, etc.

[0127] [Corrected according to Rule 26 25.11.2025] Pharmaceutically acceptable base addition salts can be formed with inorganic acids and / or organic acids. Salts derived from inorganic acids include, but are not limited to, salts of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts derived from organic acids include, but are not limited to, salts of acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, benzoic acid, succinic acid, mandelic acid, fumaric acid, malic acid, tartaric acid, citric acid, palmitic acid, 3-hydroxyl-2-naphthoic acid, salicylic acid, isothionic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. For example, the inorganic acid is hydrochloric acid. For example, the organic acid is acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, benzoic acid, succinic acid, mandelic acid, fumaric acid, malic acid, tartaric acid, citric acid, palmitic acid, 3-hydroxyl-2-naphthoic acid, salicylic acid, isothionic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like.

[0128] [Corrected according to Rule 26 25.11.2025] As used herein, the term “isomer” means different compounds having the same molecular formula. “Stereoisomers” are isomers that differ only in the way the atoms are arranged in space. “Atropisomers” are stereoisomers that result from hindered rotation about a single bond. “Enantiomers” are a pair of non-superimposable mirror images of a stereoisomer. A mixture of enantiomers in any ratio can be referred to as a “racemic” mixture. “Diastereomers” are stereoisomers that have at least two asymmetric carbons that are not mirror images of each other. “Tautomers” refer to isomeric forms of a compound that are in equilibrium with each other. The concentration of the isomeric forms will depend on the environment in which the compound is found, and can vary depending on, for example, whether the compound is a solid or in an organic or aqueous solution.

[0129] [Corrected according to Rule 26 25.11.2025] In certain embodiments, “stereoisomer” can also include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof.

[0130] [Corrected according to Rule 26 25.11.2025] The term "nucleic acid" as used herein refers generally to any compound comprising a polymer of deoxyribonucleotides (deoxyribonucleic acid, DNA for short) or a polymer of ribonucleotides (ribonucleic acid, RNA for short) or a combination thereof. In addition, nucleic acids herein also include derivatives of nucleic acids. The term "derivative of a nucleic acid" includes chemical derivatization of a nucleic acid on the base, sugar or phosphate of the nucleotide, as well as nucleic acids containing non-natural nucleotides and nucleotide analogs. Furthermore, in the present context, nucleic acids can be in the form of single- or double-stranded linear or covalently closed circular molecules.

[0131] [Corrected according to Rule 26 25.11.2025] The term "plasmid" as used herein refers generally to a circular DNA molecule, but the term can also encompass linearized DNA molecules. In particular, the term "plasmid" also encompasses molecules obtained by linearizing a circular plasmid, e.g. by digesting the circular plasmid with a restriction enzyme, thereby transforming the circular plasmid molecule into a linear molecule. Plasmids can replicate, i.e. amplify in a cell independently of the genetic information stored as chromosomal DNA, and can be used for cloning, i.e. for amplifying genetic information in bacterial cells. In one alternatively specific example, the DNA plasmid is a medium- or high-copy plasmid. In another alternatively specific example the DNA plasmid is a high-copy plasmid. Examples of such high-copy plasmids include, e.g., pUC and pTZ plasmids or any other plasmid comprising an origin of replication supporting high-copy plasmids (e.g. pMB1, pCoIE1).

[0132] [Corrected according to Rule 26 25.11.2025] The term "vaccine" as used herein is typically understood to provide at least one antigen or prophylactic or therapeutic material having the function of an antigen, which can stimulate the adaptive immune system of the body to provide an adaptive immune response.

[0133] [Corrected according to Rule 26 25.11.2025] The term "treatment" and the like, as used herein, are used in the present context to generally mean obtaining a desired pharmacological and / or physiologic effect. Thus, treatment according to the present application can relate to the treatment of the state of a disease, but can also relate to a prophylactic treatment in the sense of a complete or partial prevention of a disease or its symptoms. Preferably, in some embodiments, the term "treatment" is to be understood as being therapeutic in the sense of a partial or complete cure of a disease and / or adverse effects and / or symptoms attributed to the disease. Treatment can also be a prophylactic or preventive treatment, i.e. a measure taken to prevent a disease, e.g. to prevent the onset of an infection and / or disease.

[0134] [Corrected according to Rule 26 25.11.2025] As used herein, the term “administering” refers to providing or giving a subject a pharmaceutical agent by any effective route. Exemplary routes of administration include, but are not limited to, one or more of the following: injection (e.g., subcutaneous, intramuscular, intradermal, intraperitoneal, intrathecal, intracerebroventricular, or intravenous), oral, intra-biliary duct, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation. When used to treat a disease, disorder, condition, or symptom thereof, administration of a substance is typically performed after the onset of the disease, disorder, condition, or symptom thereof. When used to prevent a disease, disorder, condition, or symptom, administration of a substance is typically performed before the onset of the disease, disorder, condition, or symptom.

[0135] [Corrected according to Rule 26 25.11.2025] As used herein, the terms “subject” and “patient” can be used interchangeably. In certain embodiments, a subject is a mammal, such as a human, a non-human primate (e.g., simian, chimpanzee, monkey, and orangutan), a domestic animal (including dogs and cats as well as livestock animals (e.g., horses, cows, pigs, sheep, and goats)), or other mammal. Other mammals include, but are not limited to, mice, rats, guinea pigs, rabbits, hamsters, and the like. In particular embodiments, a subject is a human. In one embodiment, a subject is a mammal (e.g., a human) having an infectious or neoplastic disease. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing an infectious or neoplastic disease.

[0136] [Corrected according to Rule 26 25.11.2025] As used herein, the term “cationic lipid” refers to a lipid that becomes positively charged when the pH is lowered below the pKa of the ionizable group of the lipid, but gradually becomes neutral at higher pH values, and the positively charged lipid is capable of binding to a negatively charged nucleic acid when the pH is below the pKa. In certain embodiments, a cationic lipid comprises a zwitterionic lipid.

[0137] [Corrected according to Rule 26 25.11.2025] As used herein, the term “pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government of the United States (e.g., the National Food and Drug Administration (CFDA), the Food and Drug Administration (FDA)) or listed in the public recognized pharmacopeia (e.g., the Chinese Pharmacopeia, the European Pharmacopeia).

[0138] [Amended pursuant to Rule 26 25.11.2025] Herein, some elements of the present application will be described which, together with the specific embodiments, are presented in a simplified form to provide a clear understanding of the present application. However, it should be understood that they can be combined in any manner and in any number to produce further embodiments. The different described examples and preferred embodiments should not be interpreted as limiting the present application to only the explicitly described embodiments. The specification should be understood to support and include embodiments combining the explicitly described embodiments with any number of disclosed and / or preferred elements. Furthermore, any arrangement and combination of the described elements in the present application should be considered as being disclosed by the specification of the present application, unless the context indicates otherwise.

[0139] [Amended pursuant to Rule 26 25.11.2025] II. PEG Lipid

[0140] [Amended pursuant to Rule 26 25.11.2025] The present disclosure provides a PEG lipid as shown in Formula (I):

[0141] [Amended pursuant to Rule 26 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein

[0142] [Amended pursuant to Rule 26 25.11.2025] R1is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0143] [Amended pursuant to Rule 26 25.11.2025] Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond,

[0144] [Amended pursuant to Rule 26 25.11.2025] Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond,

[0145] [Amended pursuant to Rule 26 25.11.2025] Q1is -C(=O)-, -OC(=O)-, or a bond,

[0146] [Amended pursuant to Rule 26 25.11.2025] Q2is -C(=O)-, or a bond,

[0147] [Amended pursuant to Rule 26 25.11.2025] Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond,

[0148] [Amended pursuant to Rule 26 25.11.2025] L1is C1-C 20 alkylene or a bond,

[0149] [Amended according to Rule 26 25.11.2025] A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond,

[0150] [Amended according to Rule 26 25.11.2025] R2is C4-C 24 hydrocarbyl,

[0151] [Amended according to Rule 26 25.11.2025] The total number of carbon atoms in L1and R2is 10-35,

[0152] [Amended according to Rule 26 25.11.2025] L2is C1-C4 hydrocarbylene,

[0153] [Amended according to Rule 26 25.11.2025] L3is C1-C4 hydrocarbylene,

[0154] [Amended according to Rule 26 25.11.2025] R3is H or C1-C3 hydrocarbyl,

[0155] [Amended according to Rule 26 25.11.2025] n is any integer from 30-60.

[0156] [Amended according to Rule 26 25.11.2025] The present disclosure also provides another PEG lipid as shown in Formula (IX):

[0157] [Amended according to Rule 26 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein

[0158] [Amended according to Rule 26 25.11.2025] R1is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0159] [Amended according to Rule 26 25.11.2025] Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond,

[0160] [Amended according to Rule 26 25.11.2025] Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond, Q1is C(=O)-, -OC(=O)-, or a bond,

[0161] [Amended according to Rule 26 25.11.2025] Q2is C(=O)- or a bond,

[0162] [Corrected according to Rule 26 25.11.2025] Q3 is -C(=0)0-, -OC(=0)-, -0-, -S-, or a bond,

[0163] [Corrected according to Rule 26 25.11.2025] L1 is C1-C 20 alkylene or a bond,

[0164] [Corrected according to Rule 26 25.11.2025] A1 is -C(=0)0-, -OC(=0)-, -OC(=0)0-, -0-, -S-, or a bond,

[0165] [Corrected according to Rule 26 25.11.2025] R2 is C4-C 24 alkyl,

[0166] [Corrected according to Rule 26 25.11.2025] The total number of carbon atoms of L1 and R2 is 10-35,

[0167] [Corrected according to Rule 26 25.11.2025] L2 is C1-C4 alkylene,

[0168] [Corrected according to Rule 26 25.11.2025] L3 is C1-C4 alkylene,

[0169] [Corrected according to Rule 26 25.11.2025] R3 is H or C1-C3 alkyl,

[0170] [Corrected according to Rule 26 25.11.2025] n is any integer from 30-60,

[0171] [Corrected according to Rule 26 25.11.2025] L1 forms a four-, five-, or six-membered ring with A1 and R2.

[0172] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1 in formula (IX) is C1-C 20 alkylene.

[0173] [Corrected according to Rule 26 25.11.2025] Further provided by the present disclosure is a PEG lipid as shown in formula (X):

[0174] [Corrected according to Rule 26 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein

[0175] [Corrected according to Rule 26 25.11.2025] R1 is H, HO-Z1-Q1-, or H-Z1-Q1-,

[0176] [Corrected according to Rule 26 25.11.2025] Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2- or a bond,

[0177] [Corrected according to Rule 26 25.11.2025] Q1is C(=O)-, -OC(=O)- or a bond,

[0178] [Corrected according to Rule 26 25.11.2025] Q2is C(=O)- or a bond,

[0179] [Corrected according to Rule 26 25.11.2025] Q3is -C(=O)O-, -OC(=O)-, -O-, -S- or a bond,

[0180] [Corrected according to Rule 26 25.11.2025] L1is C1-C 20 alkylene or a bond,

[0181] [Corrected according to Rule 26 25.11.2025] A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S- or a bond,

[0182] [Corrected according to Rule 26 25.11.2025] R2is C4-C 24 alkyl,

[0183] [Corrected according to Rule 26 25.11.2025] The total number of carbon atoms of L1and R2is 10-35,

[0184] [Corrected according to Rule 26 25.11.2025] L2is C1-C4alkylene,

[0185] [Corrected according to Rule 26 25.11.2025] L3is C1-C4alkylene,

[0186] [Corrected according to Rule 26 25.11.2025] R3is H or C1-C3alkyl,

[0187] [Corrected according to Rule 26 25.11.2025] n is any integer from 30 to 60,

[0188] [Corrected according to Rule 26 25.11.2025] Z2forms a four-, five- or six-membered ring with Q3, L1, A1and R2.

[0189] [Rule 26 correction 25.11.2025] In some embodiments, in Formula (X), Z1is -L2-CH(OR3)-L3-, -L2-C(=0)-L3-, -L2-CH(R3)-L3-, -L2- or a bond, and Z2is -L2-CH(OR3)-L3-, -L2-C(=0)-L3-, -L2-CH(R3)-L3- or -L2-.

[0190] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has a structure as represented by Formula (II):

[0191] [Rule 26 correction 25.11.2025] wherein A1, L1, R1, R2, and n are as defined for Formula (I), and R1is not H and is not unsubstituted methyl.

[0192] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has a structure as represented by Formula (III):

[0193] [Rule 26 correction 25.11.2025] wherein A1, L1, R1, R2, and n are as defined for Formula (I).

[0194] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has a structure as represented by Formula (IV):

[0195] [Rule 26 correction 25.11.2025] wherein A1, L1, R1, R2, and n are as defined for Formula (I).

[0196] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has a structure as represented by Formula (V):

[0197] [Rule 26 correction 25.11.2025] wherein A1, L1, R1, R2, and n are as defined for Formula (I).

[0198] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has a structure as represented by Formula (VI):

[0199] [Rule 26 correction 25.11.2025] wherein A1, L1, R1, R2, and n are as defined for Formula (I).

[0200] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown as formula (I) has a structure as shown as formula (VII):

[0201] [Corrected according to Rule 26 25.11.2025] wherein A1, L1, R1, R2, Q2, Z2, Q3, and n are as defined for formula (I).

[0202] [Corrected according to Rule 26 25.11.2025] The PEG lipid shown as formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX), or formula (X) includes one or more of the following features, where applicable.

[0203] [Corrected according to Rule 26 25.11.2025] In some embodiments, the salt is a pharmaceutically acceptable salt.

[0204] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H or a C1-C9 hydrocarbyl group.

[0205] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H, a C1-C9 alkyl group, a C2-C9 alkenyl group, or a C2-C9 alkynyl group.

[0206] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is not H and is not unsubstituted methyl.

[0207] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is substituted methyl or is a C2-C9 hydrocarbyl group.

[0208] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is substituted methyl or is a C2-C6 hydrocarbyl group.

[0209] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is substituted methyl, is a C2-C6 alkyl group, is a C2-C6 alkenyl group, or is a C2-C6 alkynyl group.

[0210] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is a substituted hydrocarbyl group. In some embodiments, R1is a substituted C1-C9 hydrocarbyl group. In some embodiments, R1is a substituted C1-C6 hydrocarbyl group.

[0211] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Z1and Q1are as defined for formula (I).

[0212] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H-Z1-Q1-, wherein Z1and Q1are as defined for formula (I), and Z1is not a bond and is not unsubstituted methylene when Q1is a bond.

[0213] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Z1and Q1are as defined for formula (I), and Z1and Q1are not simultaneously a bond.

[0214] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Q1is as defined for formula (I), and Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3- or -L2-.

[0215] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3- or -L2-, and Q1is -C(=O)-, -OC(=O)- or a bond. In some preferred embodiments, L2is C1-C3alkylene, L3is C1-C3alkylene, and R3is H or C1-C3alkyl. Preferably, each of L2and L3is a straight chain C1-C3alkylene, more preferably C1alkylene or C2alkylene. In more preferred embodiments, R1is

[0216] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H-Z1-Q1-, Z1is -L2-, and Q1is -C(=O)-. In some preferred embodiments, L2is C1-C3alkylene. In some preferred embodiments, R1is

[0217] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H, wherein Q1, L2, L3and R3are as defined for formula (I).

[0218] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H,

[0219] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is

[0220] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is

[0221] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H or HO-Z1-Q1-. In some embodiments, R1is H or

[0222] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, Z1is -L2-, -L2-CH(OR3)-L3-, -L2-C(=O)-L3- or -L2-CH(R3)-L3-, and Q1is -OC(=O)-.

[0223] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Z1is -L2-, Q1is -OC(=O)-, and L2is as defined in formula (I).

[0224] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Z1is -L2-, Q1is -OC(=O)-, and L2is C1-C3alkylene.

[0225] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1-, wherein Z1is -L2-, Q1is -OC(=O)-, and L2is C1-C3alkylene or C2-C3alkenylene.

[0226] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H.

[0227] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O- or a bond.

[0228] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0229] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -C(=O)O-, -OC(=O)-, -O-, or a bond.

[0230] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -O- or a bond.

[0231] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is a bond.

[0232] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -C(=O)O-.

[0233] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1is -OC(=O)-.

[0234] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q2is a bond or -C(=O)-.

[0235] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q2is a bond.

[0236] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q3is a bond.

[0237] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q2and Q3are bonds.

[0238] [Corrected according to Rule 26 25.11.2025] In some embodiments, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-.

[0239] [Corrected according to Rule 26 25.11.2025] In some embodiments, Z2is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, or -L2-CH(R3)-L3-.

[0240] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2is independently, at each occurrence, C1-C3alkylene, L3is independently, at each occurrence, C1-C3alkylene, and R3is independently, at each occurrence, H or C1-C3alkyl.

[0241] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2is, at each occurrence, independently C1-C3 hydrocarbylene, preferably C1-C3 alkylene, more preferably C1alkylene or C2alkylene.

[0242] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3is, at each occurrence, independently C1-C3 hydrocarbylene, preferably C1-C3 alkylene, more preferably C1alkylene or C2alkylene.

[0243] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3is, at each occurrence, independently H or C1-C3 alkyl.

[0244] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has a structure as shown in formula (II-1):

[0245] [Corrected according to Rule 26 25.11.2025] wherein A1is -OC(=O)-, -OC(=O)O-, -O-, or -S-, L1, R2, and n are as defined in formula (II).

[0246] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has a structure as shown below:

[0247] [Corrected according to Rule 26 25.11.2025] wherein L1, R2, and n are as defined in formula (II).

[0248] [Corrected according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is not H and not unsubstituted methyl, and A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0249] [Corrected according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is HO-Z1-Q1-, and A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-, wherein Z1and Q1are as defined in formula (I), and Z1and Q1are not simultaneously a bond.

[0250] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is H-Z1-Q1-, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-, wherein Z1and Q1are as defined for formula (I), and when Q1is a bond, Z1is not a bond or Z1is a substituted methylene.

[0251] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is H-Z1-Q1-, Q1is -C(=O)-, Z1is -L2-, and A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0252] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), A1is a bond, and R1is not H and is not unsubstituted methyl.

[0253] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is HO-Z1-Q1-, A1is a bond, and Z1and Q1are not both a bond. In some such embodiments, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-. In more preferred embodiments, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-, and Q1is -C(=O)-, -OC(=O)-, or a bond. In more preferred embodiments, L2is C1-C3alkylene, L3is C1-C3alkylene, and R3is H or C1-C3alkyl. In further more preferred embodiments, L2and L3are each C1-C3alkylene, more preferably straight chain C1-C3alkylene, further more preferably C1alkylene or C2alkylene. In even more preferred embodiments, R1is

[0254] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II), R1is H-Z1-Q1-, Z1is -L2-, Q1is -C(=O)-, and A1is a bond. In some such embodiments, L2is C1-C3alkylene, preferably C1-C3alkylene, more preferably C1alkylene or C2alkylene. In more preferred embodiments, R1is

[0255] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the structure:

[0256] [Corrected according to Rule 26 25.11.2025] wherein Z1, Q1, L1, R2, and n are as defined for formula (II), and Z1and Q1are not both a bond.

[0257] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II-2) has the structure:

[0258] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II-2) has the structure:

[0259] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the structure:

[0260] [Corrected according to Rule 26 25.11.2025] wherein Z1, Q1, L1, R2, and n are as defined for formula (II), and Z1is not a bond when Q1is a bond and is not unsubstituted methylene.

[0261] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the structure:

[0262] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, Q1, R2, and n are as defined for formula (II). In some embodiments, the PEG lipid of formula (II) has the structure:

[0263] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, Q1, R2, and n are as defined for formula (II).

[0264] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the structure:

[0265] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, Q1, R2, R3, and n are as defined for formula (II).

[0266] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the following structure:

[0267] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, Q1, R2, R3, and n are as defined for formula (II).

[0268] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the following structure:

[0269] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, Q1, R2, and n are as defined for formula (II).

[0270] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q1 of the PEG lipid of formula (II-2-1) is -C(=O)-.

[0271] [Corrected according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of formula (II-2-1), when Q1 is -C(=O)-, L3 is a C1 alkylene or a C4 alkylene.

[0272] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the following structure:

[0273] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, and n are as defined for formula (II).

[0274] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the following structure:

[0275] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, R2, and n are as defined for formula (II).

[0276] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (II) has the following structure:

[0277] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, R2, and n are as defined for formula (II).

[0278] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0279] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (II).

[0280] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0281] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (II).

[0282] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0283] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (II). In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0284] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (II).

[0285] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0286] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, R2, and n are as defined for formula (II).

[0287] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid shown in formula (II) has the following structure:

[0288] [Corrected according to Rule 26 25.11.2025] wherein,

[0289] [Corrected according to Rule 26 25.11.2025] R2and L1are as defined for formula (II),

[0290] [Amended according to Rule 26 25.11.2025] (i) A1is a bond, and R2and n satisfy the following conditions: (1) the total number of carbon atoms of L1and R2is 5-12, 10-12, 14-16, 18, or 20-32; and (2) n is any integer from 36-54; or

[0291] [Amended according to Rule 26 25.11.2025] (ii) A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0292] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid represented by formula (II-4), A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-, R2and n are as defined in formula (I), and one or more of the following conditions are satisfied: (1) R2is C4-C 17 hydrocarbyl or C 19 -C 24 hydrocarbyl; and (2) the number of main chain carbon atoms of L1and the total number of carbon atoms of R2is 5-19 or 21-32.

[0293] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid represented by formula (III), R1is H or HO-Z1-Q1-, wherein Z1and Q1are as defined in formula (I), and Z1and Q1are not simultaneously a bond, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or a bond.

[0294] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid represented by formula (III) has the following structure:

[0295] [Amended according to Rule 26 25.11.2025] wherein R1, L1, R2, and n are as defined in formula (III).

[0296] [Amended according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid represented by formula (III), R1is H or HO-Z1-Q1-, wherein Z1is -L2-, Q1is -OC(=O)-, and A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or a bond.

[0297] [Amended according to Rule 26 25.11.2025] In some embodiments, the PEG lipid represented by formula (III) has the following structure:

[0298] [Rule 26 correction 25.11.2025] wherein L1, R2, and n are as defined for formula (III).

[0299] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid shown in formula (III) has the following structure:

[0300] [Rule 26 correction 25.11.2025] wherein L1, R2, and n are as defined for formula (III).

[0301] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid shown in formula (III) has the following structure:

[0302] [Rule 26 correction 25.11.2025] wherein L1, R2, and n are as defined for formula (III).

[0303] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid shown in formula (III) has the following structure:

[0304] [Rule 26 correction 25.11.2025] wherein L1, R2, and n are as defined for formula (III).

[0305] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid shown in formula (III) has the following structure:

[0306] [Rule 26 correction 25.11.2025] wherein L1, R2, and n are as defined for formula (III).

[0307] [Rule 26 correction 25.11.2025] In some embodiments, the PEG lipid shown in formula (III) has the following structure:

[0308] [Rule 26 correction 25.11.2025] wherein L1, L2, R2, and n are as defined for formula (III).

[0309] [Rule 26 correction 25.11.2025] In some embodiments, in the PEG lipid shown in formula (IV), formula (V), or formula (VI), R1is H, and A1is a bond, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, or -S-.

[0310] [Corrected according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of Formula (IV), Formula (V), or Formula (VI), R1is HO-Z1-Q1- or H-Z1-Q1-, and A1is a bond, -C(=O)O-, -OC(=O)-, or -O-.

[0311] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of Formula (IV) has the following structure:

[0312] [Corrected according to Rule 26 25.11.2025] wherein L1, R2, and n are as defined for Formula (IV).

[0313] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of Formula (V) has the following structure:

[0314] [Corrected according to Rule 26 25.11.2025] wherein R2and n are as defined for Formula (V).

[0315] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of Formula (VI) has the following structure:

[0316] [Corrected according to Rule 26 25.11.2025] wherein R2and n are as defined for Formula (VI).

[0317] [Corrected according to Rule 26 25.11.2025] The PEG lipid of Formula (VII), when applicable, includes one or more of the following features.

[0318] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1- or H-Z1-Q1-, and A1is a bond, -C(=O)O-, -OC(=O)-, or -O-.

[0319] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1- or H-Z1-Q1-, and Q2and Q3are a bond.

[0320] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1- or H-Z1-Q1-, Q2and Q3are a bond, and A1is -OC(=O)-.

[0321] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1- or H-Z1-Q1-, Q2and Q3are a bond, A1is -OC(=O)-, and Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, or -L2-CH(R3)-L3-.

[0322] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is HO-Z1-Q1- or H-Z1-Q1-, Q2and Q3are a bond, A1is -OC(=O)-, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, or -L2-CH(R3)-L3-, and L1is a bond or C1alkylenyl.

[0323] [Corrected according to Rule 26 25.11.2025] In some embodiments, R1is H, and A1is a bond, -C(=O)O-, -OC(=O)-, -OC(=O)O-, or -O-.

[0324] [Corrected according to Rule 26 25.11.2025] In some embodiments, Q2and Q3are a bond, R1is H, and Z2is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, or -L2-CH(R3)-L3-.

[0325] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (VII) has the following structure:

[0326] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (VII).

[0327] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (VII) has the following structure:

[0328] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, R3, and n are as defined for formula (VII).

[0329] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid of formula (VII) has the following structure:

[0330] [Corrected according to Rule 26 25.11.2025] wherein L1, L2, L3, R2, and n are as defined for formula (VII).

[0331] [Corrected according to Rule 26 25.11.2025] In some embodiments, in the PEG lipid of Formula (I), Y is a bond, i.e., the PEG lipid of Formula (I) has the following structure:

[0332] [Corrected according to Rule 26 25.11.2025] wherein R1, A1, L1, R2, and n are as defined for Formula (I), and

[0333] [Corrected according to Rule 26 25.11.2025] In some embodiments, when R1is H and A1is -O-, L1and R2satisfy at least one of the following conditions: (1) the total number of carbon atoms is not 16, (2) at least one of L1and R2is branched, and (3) L1is C3-C8alkylene or a bond.

[0334] [Corrected according to Rule 26 25.11.2025] In some embodiments, A1of the PEG lipid of Formula (VIII) is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -S-, or a bond.

[0335] [Corrected according to Rule 26 25.11.2025] The PEG lipids of Formula (I), Formula (II), Formula (II-1), Formula (II-1-1), Formula (II-2), Formula (II-2-1), Formula (II-2-1-a), Formula (II-2-2), Formula (II-2-2-a), Formula (II-2-2-b), Formula (II-2-3), Formula (II-2-3-a), Formula (II-2-3-b), Formula (II-2-3-c), Formula (II-2-4), Formula (II-2-4-a), Formula (II-3), Formula (II-3-1), Formula (II-3-1-a), Formula (II-4), Formula (III), Formula (III-A), Formula (III-1), Formula (III-2), Formula (III-3), Formula (III-4), Formula (III-5), Formula (III-6), Formula (IV), Formula (IV-1), Formula (V), Formula (V-1), Formula (VI), Formula (VI-1), Formula (VII), Formula (VII-1), Formula (VII-2), Formula (VII-3), Formula (VIII), Formula (IX), or Formula (X) include one or more of the following features when applicable.

[0336] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms of L1and R2is 12-25. For example, the total number of carbon atoms of L1and R2is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23.

[0337] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 11-22.

[0338] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 14-24.

[0339] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 16-22.

[0340] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 15-20.

[0341] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 16-20.

[0342] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 10-18.

[0343] [Corrected according to Rule 26 25.11.2025] In some embodiments, the total number of carbon atoms in L1and R2is 14-16. In some embodiments, L1is C1-C8alkylene or a bond.

[0344] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is C1-C 20 alkylene, C2-C 20 alkenylene, or C2-C 20 alkynylene.

[0345] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is C1-C8alkylene, C2-C8alkenylene, or C2-C8alkynylene.

[0346] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is substituted or unsubstituted C1-C 20 alkylene, e.g., substituted or unsubstituted C1-C 15 alkylene, C1-C8alkylene, C4-C8alkylene, C1-C4alkylene, or C1-C2alkylene.

[0347] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is substituted or unsubstituted C1-C 20 alkylene, e.g., substituted or unsubstituted C1-C 15alkylene, C1-C8alkylene, C4-C8alkylene, C1-C4alkylene, or C1-C2alkylene.

[0348] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is a straight chain or branched C1-C 20 alkylene, e.g., straight chain or branched C1-C 15 alkylene, C1-C8alkylene, C4-C8alkylene, C1-C4alkylene, or C1-C2alkylene.

[0349] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is a straight chain or branched C1-C 20 alkylene, e.g., straight chain or branched C1-C 15 alkylene, C1-C8alkylene, C4-C8alkylene, C1-C4alkylene, or C1-C2alkylene.

[0350] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is a straight chain C1-C 20 alkylene, e.g., straight chain C1-C 15 alkylene, C1-C8alkylene, C4-C8alkylene, C1-C4alkylene, or C1-C2alkylene.

[0351] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is a bond.

[0352] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is C4-C 24 alkyl, C4-C 24 alkenyl, or C4-C 24 alkynyl.

[0353] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is substituted or unsubstituted C4-C 24 alkyl, e.g., substituted or unsubstituted C7-C 22 alkyl, C 11 -C 22 alkyl, C4-C 20 alkyl, C8-C 20 alkyl, C 15 -C 20 alkyl, C 12 -C 18 alkyl, C 10 -C 16 alkyl, C4-C 15 alkyl, C8-C15 hydrocarbyl or C5-C 11 hydrocarbyl.

[0354] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a substituted or unsubstituted C4-C 24 alkyl, for example a substituted or unsubstituted C7-C 22 alkyl, C 11 -C 22 alkyl, C4-C 20 alkyl, C8-C 20 alkyl, C 15 -C 20 alkyl, C 12 -C 18 alkyl, C 10 -C 16 alkyl, C4-C 15 alkyl, C8-C 15 alkyl or C5-C 11 alkyl.

[0355] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a linear or branched C4-C 24 hydrocarbyl, for example a linear or branched C7-C 22 hydrocarbyl, C 11 -C 22 hydrocarbyl, C4-C 20 hydrocarbyl, C8-C 20 hydrocarbyl, C 15 -C 20 hydrocarbyl, C 12 -C 18 hydrocarbyl, C 10 -C 16 hydrocarbyl, C4-C 15 hydrocarbyl, C8-C 15 hydrocarbyl or C5-C 11 hydrocarbyl.

[0356] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a linear or branched C4-C 24 alkyl, for example a linear or branched C7-C 22 alkyl, C 11 -C 22 alkyl, C4-C 20 alkyl, C8-C 20 alkyl, C 15 -C 20 alkyl, C 12 -C 18 alkyl, C 10 -C16 alkyl, C4-C 15 alkyl, C8-C 15 alkyl or C5-C 11 alkyl.

[0357] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a linear C4-C 24 alkyl, e.g. linear C7-C 22 alkyl, linear C 11 -C 22 alkyl, linear C4-C 20 alkyl, linear C8-C 20 alkyl, linear C 15 -C 20 alkyl, linear C 12 -C 18 alkyl, linear C 10 -C 16 alkyl, linear C4-C 15 alkyl, linear C8-C 15 alkyl or linear C5-C 11 alkyl.

[0358] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a C7-C 22 alkyl.

[0359] [Corrected according to Rule 26 25.11.2025] In some embodiments, R2is a C7-C 22 alkyl, C7-C 22 alkenyl or C7-C 22 alkynyl.

[0360] [Corrected according to Rule 26 25.11.2025] In some embodiments, L1is a linear C1-C8alkylene, and R2is a linear C7-C 22 alkyl.

[0361] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2is a linear C 16 -C 22 alkyl.

[0362] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2is a C 16 -C 22 alkyl.

[0363] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2is a linear C16 ~ C 22 alkyl.

[0364] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2 is C 16 ~ C 18 alkyl.

[0365] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2 is straight chain C 16 ~ C 18 alkyl.

[0366] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2 is C 16 ~ C 18 alkyl.

[0367] [Corrected according to Rule 26 25.11.2025] In some embodiments, -L1-R2 is straight chain C 16 ~ C 18 alkyl.

[0368] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is H, C1-C3 alkyl, C2-C3 alkenyl, or C2-C3 alkynyl.

[0369] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is H or C1-C3 alkyl.

[0370] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C1-C3 alkyl, C2-C3 alkenyl, or C2-C3 alkynyl.

[0371] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C1-C2 alkyl.

[0372] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C1 alkyl or C2 alkyl.

[0373] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C1 alkyl or C2 alkyl.

[0374] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is H.

[0375] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C1 alkyl.

[0376] [Corrected according to Rule 26 25.11.2025] In some embodiments, R3 is C2alkyl.

[0377] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 30 to 55, for example, n is any integer from 30 to 50, 34 to 50, 34 to 55, 36 to 55, 40 to 55, 44 to 55, 48 to 55, 50 to 55, 36 to 54, 40 to 54, 44 to 54, 48 to 54, or 50 to 54.

[0378] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 35 to 55.

[0379] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 35 to 50.

[0380] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is any integer from 40 to 50.

[0381] [Corrected according to Rule 26 25.11.2025] In some embodiments, n is 45.

[0382] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2 is C1-C3 hydrocarbylene.

[0383] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2 is C1-C3 alkylene, C2-C3 alkenylene, or C2-C3 alkynylene.

[0384] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2 is C1-C3 alkylene.

[0385] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2 is linear C1-C3 alkylene.

[0386] [Corrected according to Rule 26 25.11.2025] In some embodiments, L2 is C1 alkylene or C2 alkylene.

[0387] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3 is C1-C2 hydrocarbylene.

[0388] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3 is C1-C2 alkylene, C2 alkenylene, or C2 alkynylene.

[0389] [Corrected according to Rule 26 25.11.2025] In some embodiments, L3 is a C1-C2 alkylene group.

[0390] [Corrected according to Rule 26 25.11.2025] In some embodiments, the PEG lipid represented by Formula (I) has the structure shown in Table 1 below:

[0391] [Corrected according to Rule 26 25.11.2025] Table 1

[0392] [Corrected according to Rule 26 25.11.2025] III. Lipid nanoparticles, pharmaceutical compositions, drugs, and uses thereof

[0393] [Corrected according to Rule 26 25.11.2025] The present disclosure also provides a lipid nanoparticle comprising the PEG lipid of any of the above embodiments.

[0394] [Corrected according to Rule 26 25.11.2025] In some embodiments, the lipid nanoparticle refers to a particle with a size in the nanometer scale (e.g., lnm-1000 nm).

[0395] [Corrected according to Rule 26 25.11.2025] In some embodiments, the average diameter of the above-mentioned lipid nanoparticle is 20-800 nm, 20-500 nm, 20-400 nm, 20-300 nm, 20-200 nm, 20-100 nm, 30-700 nm, 30-500 nm, 30-300 nm, 30-200 nm, 30-100 nm, 40-800 nm, 40-600 nm, 40-500 nm, 40-300 nm, 40-200 nm, 40-100 nm, 50-800 nm, 50-600 nm, 50-500 nm, 50-400 nm, 50-300 nm, 50-200 nm, 50-100 nm, 60-800 nm, 60-600 nm, 60-500 nm, 60-400 nm, 60-300 nm, 60-200 nm, or 60-100 nm. In some alternative specific examples, the average diameter of the above-mentioned lipid nanoparticle is 26 nm, 31 nm, 36 nm, 41 nm, 46 nm, 51 nm, 56 nm, 61 nm, 66 nm, 71 nm, 76 nm, 81 nm, 86 nm, 91 nm, 96 nm, 101 nm, 106 nm, 111 nm, 116 nm, 121 nm, 126 nm, 131 nm, 136 nm, 141 nm, 146 nm, 151 nm, 156 nm, 161 nm, 166 nm, 171 nm, 176 nm, 181 nm, 186 nm, 191 nm, 196 nm, 201 nm, 206 nm, 211 nm, 216 nm, 221 nm, 226 nm, 231 nm, 236 nm, 241 nm, 246 nm, or 249 nm. In this context, the average diameter of the lipid nanoparticle can be represented by the z-average value determined by dynamic light scattering.

[0396] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-mentioned lipid nanoparticle comprises one of the following: cationic lipid nanoparticle, solid lipid nanoparticle (SLN), nanostructured lipid carrier (NLC), non-lamellar lipid nanoparticle. In one alternative specific example, the above-mentioned lipid nanoparticle is a cationic lipid nanoparticle.

[0397] [Rule 26 correction 25.11.2025] In some embodiments, the above-mentioned lipid nanoparticle further contains one or more of the following: a cationic lipid, a helper lipid, and a structural lipid.

[0398] [Rule 26 correction 25.11.2025] In some embodiments, the cationic lipid comprises a compound of Formula (XI), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0399] [Rule 26 correction 25.11.2025] wherein,

[0400] [Rule 26 correction 25.11.2025] R 1 is selected from the group consisting of C5-C12alkyl, C5-C12alkenyl, -R*Y1R”, -Y1R”, and -R”’M’R’; 30 is selected from the group consisting of C5-C12alkyl, C5-C12alkenyl, -R*Y1R”, -Y1R”, and -R”’M’R’; 20 is selected from the group consisting of C5-C12alkyl, C5-C12alkenyl, -R*Y1R”, -Y1R”, and -R”’M’R’;

[0401] [Rule 26 correction 25.11.2025] R 2 and R 3 are independently selected from the group consisting of H, C1-C6alkyl, C2-C6alkenyl, -R*Y1R”, -Y1R”, and -R*OR”, or R 14 and R 14 are independently selected from the group consisting of H, C1-C6alkyl, C2-C6alkenyl, -R*Y1R”, -Y1R”, and -R*OR”, or R 2 and R 3 together with the atoms to which they are attached form a heterocycle or a carbocycle;

[0402] [Rule 26 correction 25.11.2025] R4is selected from the group consisting of hydrogen, C3-C6carbocycle, -(CH2) n Q, -(CH2) n CHQR, -(CH2) o C(R 10 )2(CH2) n-o Q, -CHQR, -CQ(R)2, and unsubstituted C1-C6alkyl, wherein Q is selected from carbocycle, heterocycle, -OR, -O(CH2) n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, N(R)R8, -N(R)S(O)2R8, -O(CH2) nOR, -N(R)C(=NR9)N(R)2, -N(R)C(=CHR9)N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, - N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N( R)2, -N(OR)C(=NR9)N(R)2, -N(OR)C(=CHR9)N(R)2, -C(=NR9)N(R)2, -C(=NR9) R, -C(O)N(R)OR and -C(R)N(R)2C(O)OR, each o is independently selected from 1, 2, 3 and 4, and each n is independently selected from 1, 2, 3, 4 and 5;

[0403] [Amended according to Rule 26, 25.11.2025] Each R5 is independently selected from the group consisting of OH, C1-C3 alkyl, C2-C3 alkenyl and H;

[0404] [Amended according to Rule 26, 25.11.2025] Each R6 is independently selected from the group consisting of OH, C1-C3 alkyl, C2-C3 alkenyl and H;

[0405] [Amended according to Rule 26, 25.11.2025] M and M' are independently selected from -C(O)O-, -OC(O)-, -OC(O)-M”-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O), -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, aryl groups and heteroaryl groups, wherein M” is a bond, C1~C 13 Alkylene or C 2- ~C 13 alkenyl;

[0406] [Revised from Rule 26 to 25.11.2025] R7 can be selected freely as C. 1-3 The group consisting of alkyl groups, C2-C3 alkenyl groups, and H;

[0407] [Revised from Rule 26 to 25.11.2025] R8 can be selected freely from C. 3-6 A group consisting of carbon rings and heterocycles;

[0408] [Amended according to Rule 26 25.11.2025] R9 is selected from the group consisting of: H, CN, NO2, C1-C6 alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C2-C6 alkenyl, C3-C6 carbon ring and heterocycle;

[0409] [Amended according to Rule 26, dated November 2025]10 selected from the group consisting of H, C1-C3 alkyl, and C2-C3 alkenyl;

[0410] [Amended according to Rule 26 25.11.2025] each R is independently selected from the group consisting of C1-C3 alkyl, C2-C3 alkenyl, (CH2) q OR**and H,

[0411] [Amended according to Rule 26 25.11.2025] and each q is independently selected from 1, 2, and 3;

[0412] [Amended according to Rule 26 25.11.2025] each R’ is independently selected from the group consisting of C1-C 18 alkyl, C2-C 18 alkenyl, -R*Y1R”, -Y1R”, H, and and R 11 is selected from the group consisting of C1-C 12 alkylene, and C2-C 12 alkenylene, R 12 and R 13 are each independently selected from the group consisting of C1-C 12 alkyl, and C2-C 12 alkenyl;

[0413] [Amended according to Rule 26 25.11.2025] each R” is independently selected from the group consisting of C3-C 15 alkyl, and C3-C 15 alkenyl;

[0414] [Amended according to Rule 26 25.11.2025] each R”’ is independently selected from the group consisting of C3-C 15 alkylene, and C3-C 15 alkenylene;

[0415] [Amended according to Rule 26 25.11.2025] each R* is independently selected from the group consisting of absent, C1-C 12 alkylene, and C2-C 12 alkenylene;

[0416] [Amended according to Rule 26 25.11.2025] each R** is independently selected from the group consisting of absent, C1-C 12 alkyl, and C2-C 12 alkenyl;

[0417] [Amended according to Rule 26 25.11.2025] each Y1 is independently C3-C6 carbocyclic;

[0418] [Amended pursuant to Rule 26 25.11.2025] Each X is independently selected from the group consisting of F, Cl, Br, and I; and

[0419] [Amended pursuant to Rule 26 25.11.2025] m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13; and wherein when R4is -(CH2) n Q, -(CH2) n CHQR, -CHQR, or -CQ(R)2, then (i) when n is 1, 2, 3, 4, or 5, Q is not -N(R)2; or (ii) when n is 1 or 2, Q is not 5-, 6-, or 7-membered heterocycloalkyl.

[0420] [Amended pursuant to Rule 26 25.11.2025] In one alternatively specific example, the cationic lipid is a compound represented by formula (XI), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0421] [Amended pursuant to Rule 26 25.11.2025] wherein, R 1 ~R 3 , R4-R7, M, and m are as defined above in formula (XI).

[0422] [Amended pursuant to Rule 26 25.11.2025] In one alternatively specific example, the cationic lipid is a compound represented by formula (XI-1), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0423] [Amended pursuant to Rule 26 25.11.2025] In some embodiments, the cationic lipid comprises a compound represented by formula (XII) below, an N-oxide thereof, a salt thereof, or an isomer thereof:

[0424] [Amended pursuant to Rule 26 25.11.2025] wherein,

[0425] [Amended pursuant to Rule 26 25.11.2025] R 1 , R 2 , R 3 , R5, R6, M, and R7 are as defined above in formula (XI),

[0426] [Amended pursuant to Rule 26 25.11.2025] R N is H or C1-C3 alkyl;

[0427] [Amended pursuant to Rule 26 25.11.2025] X a and X b are each independently O or S;

[0428] [Corrected according to Rule 26 25.11.2025] R 14 selected from the group consisting of H, halogen, -OH, R b , -N(R b )2, -CN, -N3, -C(O)OH, -C(O)OR b , -OC(O)R b , -OR b , -SR b , -S(O)R b , -S(O)OR b , -S(O)2OR b , -NO2, -S(O)2N(R b )2, -N(R b )S(O)2R b , -NH(CH2) t1 N(R b )2, -NH(CH2) p1 O(CH2) q1 N(R b )2, -NH(CH2) s1 OR b , -N((CH2) s1 OR b )2, -N(R b )-carbocyclic, -N(R b )-heterocyclic, -N(R b )-aryl, -N(R b )-heteroaryl, -N(R b )(CH2) t1 -carbocyclic, -N(R b )(CH2) t1 -heterocyclic, -N(R b )(CH2) t1- -aryl, -N(R b )(CH2) t1 -heteroaryl, carbocyclic, heterocyclic, aryl and heteroaryl;

[0429] [Corrected according to Rule 26 25.11.2025] each R b is independently selected from the group consisting of C1-C3 alkyl, C2-C3 alkenyl and H;

[0430] [Corrected according to Rule 26 25.11.2025] u is 5, 6, 7, 8, 9, 10, 11, 12 or 13;

[0431] [Corrected according to Rule 26 25.11.2025] w is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0432] [Amended according to Rule 26 25.11.2025] r is 0 or 1 ;

[0433] [Amended according to Rule 26 25.11.2025] t 1 is 1, 2, 3, 4, or 5;

[0434] [Amended according to Rule 26 25.11.2025] p 1 is 1, 2, 3, 4, or 5;

[0435] [Amended according to Rule 26 25.11.2025] q 1 is 1, 2, 3, 4, or 5; and

[0436] [Amended according to Rule 26 25.11.2025] s 1 is 1, 2, 3, 4, or 5.

[0437] [Amended according to Rule 26 25.11.2025] In one optional embodiment, the cationic lipid is a compound of Formula (XII), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0438] [Amended according to Rule 26 25.11.2025] wherein R 1 ~ R 3 , R5~R7, R 14 , X a , X b , R N , M, u, w, and r are as defined above for Formula (XII).

[0439] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid comprises a compound of Formula (XIII), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0440] [Amended according to Rule 26 25.11.2025] wherein:

[0441] [Amended according to Rule 26 25.11.2025] L 1 or L 2 is -0(C=0)-, -(C=0)0-, -C(=0)-, -0-, -S(O) x -, -S-S-, -C(=0)S-, -SC(=0)-, -NR a C(=0)-, -C(=0)NR a -, NR a C(=0)NR a-OC(=O)NR a - or -NR a C(=O)O-, and L 1 or L 2 the other one is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -C(=O)S-, SC(=O)-, -NR a C(=O)-, -C(=O)NR a -, NR a C(=O)NR a -, -OC(=O)NR a - or -NR a C(=O)O- or a bond;

[0442] [Amended according to Rule 26 25.11.2025] G 1 and G 2 each independently unsubstituted C1-C 12 alkylene or C2-C 12 alkenylene;

[0443] [Amended according to Rule 26 25.11.2025] G 3 is C1-C 24 alkylene, C2-C 24 alkenylene, C3-C8cycloalkylene, C3-C8cycloalkenylene;

[0444] [Amended according to Rule 26 25.11.2025] R a is H or C1-C 12 alkyl;

[0445] [Amended according to Rule 26 25.11.2025] R 15 and R 16 each independently C6-C 24 alkyl or C6-C 24 alkenyl;

[0446] [Amended according to Rule 26 25.11.2025] R 17 is H, OR 18 , CN, -C(=O)OR 19 , -OC(=O)R 19 or -NR 18 C(=O)R 19 ;

[0447] [Amended according to Rule 26 25.11.2025] R 19 is C1-C 12 alkyl;

[0448] [Amended according to Rule 26 25.11.2025] R 18 is H or C1-C6alkyl; and

[0449] [Amended according to Rule 26 25.11.2025] x is 0, 1, or 2.

[0450] [Amended according to Rule 26 25.11.2025] In an alternative embodiment, the cationic lipid is a compound of Formula (XIII), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0451] [Amended according to Rule 26 25.11.2025] wherein: R 15 ~ R 17 , G 1 ~ G 3 , L 1 ~ L 2 is as defined for Formula (XIII) above.

[0452] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid comprises a compound of Formula (XIV), or a pharmaceutically acceptable salt or stereoisomer thereof:

[0453] [Amended according to Rule 26 25.11.2025] wherein:

[0454] [Amended according to Rule 26 25.11.2025] L 3 and L 4 are the same or different, each independently C1-C 12 alkylene, C2-C 12 alkenylene, or C2-C 12 alkynylene; in some embodiments L 3 and L 4 are the same or different, each independently C3-C 10 alkylene, C3-C 10 alkenylene, or C3-C 10 alkynylene; in some embodiments, L 3 and L 4 are the same or different, each independently C3-C 10 alkylene; in some embodiments, L 3 and L 4 are the same or different, each independently C5-C8alkylene;

[0455] [Amended according to Rule 26 25.11.2025] G 4and G 5 are the same or different, each independently -0-(C=0)-, -(C=0)-0-, -C(=0)-, -0-, -C(=0)-S-, or -S-C(=0)-; in some embodiments, G 4 and G 5 are the same or different, each independently -0-(C=0)-, -(C=0)-0-, -C(=0)-, or -0-; in some embodiments, G 4 and G 5 are the same or different, each independently selected from -0-(C=0)- or -(C=0)-0-;

[0456] [Rule 26 correction 25.11.2025] R 18 and R 19 are the same or different, each independently C5-C12alkyl or C5-C12alkenyl containing one or more double bonds; 27 are the same or different, each independently C5-C12alkyl or C5-C12alkenyl containing one or more double bonds; 27 are the same or different, each independently C8-C12alkyl or C8-C12alkenyl containing one or more double bonds; in some embodiments, R 18 and R 19 are the same or different, each independently C8-C12alkyl or C8-C12alkenyl containing one or more double bonds; in some embodiments, R 20 are the same or different, each independently C9-C12alkyl or C9-C12alkenyl containing one or two double bonds; in some embodiments, R 20 are the same or different, each independently C9-C12alkyl or C9-C12alkenyl containing one or two double bonds; in some embodiments, R 18 and R 19 are the same or different, each independently C9-C12alkyl or C9-C12alkenyl containing one or two double bonds; in some embodiments, R 17 are the same or different, each independently C9-C12alkyl or C9-C12alkenyl containing one or two double bonds; in some embodiments, R 18 are the same or different, each independently C9-C12alkyl or C9-C12alkenyl containing one or two double bonds; in some embodiments, R 18 and R 19 are the same or different, each independently

[0457] [Rule 26 correction 25.11.2025] R 20 is halogen, hydroxy, cyano, C1-C6alkyl, nitro, C1-C6alkoxy, C1-C6alkylcarbonyloxy, C1-C6alkoxycarbonyl, C1-C6alkylaminocarbonyl, or C1-C6alkylcarbonylamino; in some embodiments, R 20 is halogen, hydroxy, cyano, C1-C6alkoxy, C1-C6alkylcarbonyloxy, C1-C6alkoxycarbonyl, C1-C6alkylaminocarbonyl, or C1-C6alkylcarbonylamino; in some embodiments, R 20 is halogen, hydroxy, cyano, C1-C4alkoxy, C1-C4alkylcarbonyloxy, C1-C4alkoxycarbonyl, C1-C4alkylaminocarbonyl, or C1-C4alkylcarbonylamino; in some embodiments, R 20fluoro, hydroxy, cyano, methoxy, acetoxy, methoxycarbonyl, butylaminocarbonyl, or acetylamino;

[0458] [Amended according to Rule 26 25.11.2025] z is 1, 2, or 3.

[0459] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid is a compound of Formula (XIV), or a pharmaceutically acceptable salt or stereoisomer thereof:

[0460] [Amended according to Rule 26 25.11.2025] wherein: R 18 ~R 20 , G 4 ~G 5 , L 3 ~L 4 and z are as defined above in Formula (XIV).

[0461] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid comprises the following compound (XIV-1), or a pharmaceutically acceptable salt or stereoisomer thereof:

[0462] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid comprises the following compound, or a pharmaceutically acceptable salt thereof:

[0463] [Amended according to Rule 26 25.11.2025] In some embodiments, the cationic lipid of the above-described lipid nanoparticle comprises one or more of ALC-0315 (CAS No. 2036272-55-4), SM-102 (CAS No. 2089251-47-6), and

[0464] [Corrected according to Rule 26 25.11.2025] In some embodiments, the helper lipid of the above-described lipid nanoparticle comprises a phospholipid. The phospholipid is typically semi-synthetic, and can also be of natural origin or chemically modified. In one alternative specific example, the helper lipid of the above-described lipid nanoparticle is a phospholipid. In some embodiments, the phospholipid of the above-described lipid nanoparticle comprises one or more of: DSPC (distearoylphosphatidylcholine), DOPE (dioleoylphosphatidylethanolamine), DOPC (dioleoylphosphatidylcholine), DOPS (dioleoylphosphatidylserine), DSPG (1,2-dioctadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol)), DPPG (dipalmitoylphosphatidylglycerol), DPPC (dipalmitoylphosphatidylcholine), DGTS (1,2-dipalmitoyl-sn-glycero-3-0-4'-(N,N,N-trimethyl)homoserine), and lysophospholipid. In some embodiments, the helper lipid of the above-described lipid nanoparticle is selected from one or more of: DSPC, DOPE, DOPC, and DOPS. In some embodiments, the helper lipid of the above-described lipid nanoparticle is DSPC and / or DOPE.

[0465] [Corrected according to Rule 26 25.11.2025] In some embodiments, the structural lipid of the above-described lipid nanoparticle comprises a sterol. In one alternative specific example, the structural lipid of the above-described lipid nanoparticle is a sterol. In some embodiments, the sterol of the above-described lipid nanoparticle comprises one or more of: 20a-hydroxycholesterol, cholesterol, cholesterol ester, sterol hormone, sterol vitamin, bile acid, cholestanol, ergosterol, beta-sitosterol, and oxidized cholesterol derivative. In some embodiments, the structural lipid of the above-described lipid nanoparticle comprises at least one of: cholesterol, cholesterol ester, sterol hormone, sterol vitamin, and bile acid. In some embodiments, the structural lipid of the above-described lipid nanoparticle is cholesterol. In one alternative specific example, the structural lipid of the above-described lipid nanoparticle is high purity cholesterol, particularly injection grade high purity cholesterol, such as CHO-HP (produced by AVT). In other embodiments, the structural lipid is 20a-hydroxycholesterol.

[0466] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-described lipid nanoparticles contain a cationic lipid, a helper lipid, a structural lipid, and a PEG lipid of any of the above-described embodiments. In some embodiments, the above-described lipid nanoparticles comprise the following amount (mole percent) of cationic lipid, based on the total amount of cationic lipid, helper lipid, structural lipid, and PEG lipid: about 25-75%, e.g., about 25-28%, 28-32%, 32-35%, 35-40%, 40-42%, 42-45%, 45-46.3%, 46.3-48%, 48-49.5%, 49.5-50%, 50-55%, 55-60%, 60-65%, or 65-75%.

[0467] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-described lipid nanoparticles comprise the following amount (mole percent) of helper lipid, based on the total amount of cationic lipid, helper lipid, structural lipid, and PEG lipid: about 5-45%, e.g., about 5-9%, 9-9.4%, 9.4-10%, 10-10.5%, 10.5-11%, 11-15%, 15-16%, 16-18%, 18-20%, 20-25%, 25-33.5%, 33.5-37%, 37-40%, 40-42%, or 42-45%.

[0468] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-described lipid nanoparticles comprise the following amount (mole percent) of structural lipid, based on the total amount of cationic lipid, helper lipid, structural lipid, and PEG lipid: about 0-50%, e.g., about 0-10%, 10-15.5%, 15.5-18.5%, 18.5-22.5%, 22.5-23.5%, 23.5-28.5%, 28.5-33.5%, 33.5-35%, 35-36.5%, 36.5-38%, 38-38.5%, 38.5-39%, 39-39.5%, 39.5-40.5%, 40.5-41.5%, 41.5-42.5%, 42.5-42.7%, 42.7-43%, 43-43.5%, 43.5-45%, 45-46.5%, 46.5-48.5%, or 46.5-50%.

[0469] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above-mentioned lipid nanoparticles comprise PEG lipid in an amount (molar percentage) of about 0.5-5%, for example about 0.5-1%, 1-1.5%, 1.5-1.6%, 1.6-2%, 2-2.5%, 2.5-3%, 3-3.5%, 3.5-4%, 4-4.5%, or 4.5-5%, based on the total amount of cationic lipid, helper lipid, structural lipid, and PEG lipid.

[0470] [Corrected according to Rule 26 25.11.2025] In some embodiments as described above, in the lipid nanoparticles the molar ratio of cationic lipid : helper lipid : structural lipid : PEG lipid is about 45:10:42.5:2.5, 45:11:41.5:2.5, 42:10.5:45:2.5, 42:16:39.5:2.5, 40:16:41.5:2.5, 40:18:39.5:2.5, 35:16:46.5:2.5, 35:25:36.5:3.5, 28:33.5:35:3.5, 32:37:40.5:0.5, 35:40:22.5:2.5, 40:42:15.5:2.5, 40:20:38.5:1.5, 45:15:38.5:1.5, 55:5:38.5:1.5, 60:5:33.5:1.5, 45:20:33.5:1.5, 50:20:28.5:1.5, 55:20:23.5:1.5, 60:20:18.5:1.5, 40:15:43.5:1.5, 50:15:33.5:1.5, 55:15:28.5:1.5, 60:15:23.5:1.5, 40:10:48.5:1.5, 45:10:43.5:1.5, 55:10:33.5:1.5, 40:5:53.5:1.5, 45:5:48.5:1.5, or 50:5:43.5:1.5. In some such embodiments, the helper lipid is DOPE and the structural lipid is CHO-HP.

[0471] [Rule 26 correction 25.11.2025] In other embodiments as described above, in the lipid nanoparticle, the molar ratio of cationic lipid: helper lipid: structural lipid: PEG lipid is about 50:10:38.5:1.5, 50:9:38:3, 49.5:10:39:1.5, 48:10:40.5:1.5, 46.3:9.4:42.7:1.6, 45:9:43:3, 45:11:41.5:2.5, 42:10.5:45:2.5, 42:16:39.5:2.5, 40:16:41.5:2.5, 40:18:39.5:2.5, 35:40:22.5:2.5, 40:20:38.5:1.5, 45:15:38.5:1.5, 55:5:38.5:1.5, 60:5:33.5:1.5, 45:20:33.5:1.5, 50:20:28.5:1.5, 55:20:23.5:1.5, 60:20:18.5:1.5, 40:15:43.5:1.5, 50:15:33.5:1.5, 55:15:28.5:1.5, 60:15:23.5:1.5, 40:10:48.5:1.5, 45:10:43.5:1.5, 55:10:33.5:1.5, 40:5:53.5:1.5, 45:5:48.5:1.5, or 50:5:43.5:1.5. In some such embodiments, the helper lipid is DSPC and the structural lipid is CHO-HP.

[0472] [Rule 26 correction 25.11.2025] In some embodiments, the lipid nanoparticle described above further comprises an active ingredient.

[0473] [Rule 26 correction 25.11.2025] In some embodiments, the active ingredient is a pharmaceutically active ingredient.

[0474] [Amended according to Rule 26 25.11.2025] In some embodiments, the active ingredient is a nucleic acid, e.g., DNA, RNA. In some embodiments, the pharmaceutically active ingredient is a nucleic acid, e.g., DNA, RNA. In some embodiments, the pharmaceutically active ingredient is one or more of: RNA and DNA. In some embodiments, the RNA is one or more of: mRNA, antisense oligonucleotide, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA), and Cas9 mRNA. In some embodiments, the pharmaceutically active ingredient is mRNA. In some embodiments, the pharmaceutically active ingredient is a plasmid.

[0475] [Amended according to Rule 26 25.11.2025] In some embodiments, the nucleic acid as a pharmaceutically active ingredient encodes a polypeptide and / or protein of interest. A polypeptide or protein of interest refers to a therapeutically or pharmaceutically active polypeptide or protein whose function in or near a cell is desirable or beneficial, e.g., a protein whose absence or defective form causes a disease, providing the protein can modulate or prevent the disease, or a protein that is beneficial to the body in or near a cell. The polypeptide or protein can comprise an intact protein or a functional variant thereof.

[0476] [Amended according to Rule 26 25.11.2025] In some embodiments, the polypeptide and / or protein expressed by the nucleic acid encoding a polypeptide and / or protein of interest comprises or is one or more of: (a) an antigen; (b) a therapeutic protein or polypeptide, fragment, fragment, or variant thereof; and (c) other polypeptides or proteins.

[0477] [Amended according to Rule 26 25.11.2025] The PEG lipid as shown in Formula (I) or the lipid nanoparticle (LNP) containing the PEG lipid as shown in Formula (I) avoids an immune response in a subject against the LNP.

[0478] [Amended according to Rule 26 25.11.2025] In some embodiments, repeated administration (e.g., 2, 3, 4, or more times) of the PEG lipid as shown in Formula (I) or the LNP containing the PEG lipid as shown in Formula (I) does not induce an immune response in a subject against the LNP.

[0479] [Corrected according to Rule 26 25.11.2025] In some embodiments, repeated administration (e.g., 2, 3, 4, or more administrations) of the PEG lipid of Formula (I) or LNPs containing the PEG lipid of Formula (I) does not induce production of IgM or IgG that binds to the LNPs. By not inducing production of IgM or IgG that binds to the LNPs is meant not inducing production of native IgM or IgG that binds to the LNPs, or inducing low levels of IgM or IgG that is not sufficient to result in ABC.

[0480] [Corrected according to Rule 26 25.11.2025] The PEG lipid of Formula (I) or LNPs containing the PEG lipid of Formula (I) reduces the immune response of a subject against the LNPs.

[0481] [Corrected according to Rule 26 25.11.2025] In some embodiments, repeated administration (e.g., 2, 3, 4, or more administrations) of LNPs containing the PEG lipid of Formula (I) induces less IgM or IgG compared to control LNPs (e.g., LNPs in which the PEG lipid is DMG-PEG, where DMG refers to 1,2-dimyristoyl-sn-glycerol, and DMG-PEG is a polyethylene glycol derivative of DMG), for example at least a 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% reduction.

[0482] [Corrected according to Rule 26 25.11.2025] The PEG lipid of Formula (I) or LNPs containing the PEG lipid of Formula (I) avoids the occurrence of accelerated blood clearance (ABC).

[0483] [Corrected according to Rule 26 25.11.2025] In some embodiments, repeated administration of the PEG lipid of Formula (I) or LNPs containing the PEG lipid of Formula (I) also does not cause the occurrence of accelerated blood clearance (ABC).

[0484] [Corrected according to Rule 26 25.11.2025] The PEG lipid of Formula (I) or LNPs containing the PEG lipid of Formula (I) reduces the extent of accelerated blood clearance (ABC).

[0485] [Corrected according to Rule 26 25.11.2025] In some embodiments, the clearance of the active ingredient is less, e.g., at least 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 100% less, when the active ingredient is delivered a second or more times using LNPs containing the PEG lipid of Formula (I) as compared to control LNPs, e.g., LNPs in which the PEG lipid is DMG-PEG.

[0486] [Corrected according to Rule 26 25.11.2025] In some embodiments, the clearance of the active ingredient is less, e.g., at least 1% to 5%, 10% to 100%, 10% to 50%, 20% to 100%, 20% to 50%, 30% to 100%, 30% to 50%, 40% to 100%, 40% to 80%, 50% to 90%, or 50% to 100% less, when the active ingredient is delivered a second or more times using LNPs containing the PEG lipid of Formula (I) as compared to control LNPs, e.g., LNPs in which the PEG lipid is DMG-PEG.

[0487] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a composition comprising the lipid nanoparticle of any of the above embodiments, and a biologically acceptable carrier. The term “biologically acceptable carrier” as used herein refers to carriers that do not have significant stimulatory effects on natural or artificial organisms, including humans, animals, plants, or tissues or cells thereof in vivo or in vitro; single-cell organisms, such as bacteria and fungi; and artificial cells, and do not impair the activity and performance of the active ingredient in the lipid nanoparticle.

[0488] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a pharmaceutical composition comprising the lipid nanoparticle of any of the above embodiments, and a pharmaceutically acceptable carrier.

[0489] [Corrected according to Rule 26 25.11.2025] The above pharmaceutical composition comprises the above lipid nanoparticle, and has the corresponding advantages thereof.

[0490] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides the use of the lipid nanoparticle of any of the above embodiments or the pharmaceutical composition of any of the above embodiments in the preparation of a medicament.

[0491] [Corrected according to Rule 26 25.11.2025] In some embodiments, the above medicament is used for the treatment and / or prevention of a disease.

[0492] [Amended according to Rule 26 25.11.2025] In some embodiments, the above drug is used for gene therapy, gene vaccination, protein replacement therapy, antisense therapy or treatment by interfering RNA.

[0493] [Amended according to Rule 26 25.11.2025] In some embodiments, the above lipid particle is used as a carrier for transferring or delivering an active ingredient. In some embodiments, the active ingredient is a pharmaceutically active ingredient, such as a nucleic acid.

[0494] [Amended according to Rule 26 25.11.2025] In some embodiments, the above drug is used for treating a disease corresponding to the above polypeptide or protein of interest.

[0495] [Amended according to Rule 26 25.11.2025] In some embodiments, the above drug is used for treating and / or preventing one or more of the following diseases: rare diseases, cancer, infectious diseases, autoimmune diseases, metabolic diseases, neurological diseases, cardiovascular diseases, transplant rejection, inflammatory reactions, genetic diseases and musculoskeletal diseases.

[0496] [Corrected according to Rule 26 25.11.2025] In some embodiments, the rare disease described above comprises one or more of the following: Osteogenesis Imperfecta (Brittle Bone Disease), Wilson Disease, Spinal Muscular Atrophy (SMA), Huntingdon’s Disease, Rett Syndrome, Amyotrophic Lateral Sclerosis (ALS), Duchenne Type Muscular dystrophy, Friedrichs Ataxia, methylmalonic acidemia (MMA), Cystic Fibrosis (CF), glycogen storage disease 1a (GSD1a), glycogen storage disease III (GSD III), Crigler-Najjar syndrome, ornithine transcarbamylase deficiency (OTCD), propionic acidemia (PA), phenylketonuria (PKU), hemophilia A, hemophilia B, β-thalassemia, Lafora disease, Dravet syndrome (DS), Alexander disease, Leber’s congenital amaurosis (LCA), Myelodysplastic Syndrome (MDS), Homocystinuria due to CBS deficiency. In addition, the rare diseases in www.orpha.net / consor / cgi-bin / Disease_Search_List.php and rarediseases.info.nih.gov / diseases are incorporated herein.

[0497] [Corrected according to Rule 26 25.11.2025] In some embodiments, the cancer described above comprises one or more of: hematological malignancies, lung cancer, liver cancer, kidney cancer, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, pancreatic cancer, brain cancer, prostate cancer, gallbladder cancer, ovarian cancer, breast cancer, cervical cancer, endometrial cancer, bladder cancer, and melanoma.

[0498] [Corrected according to Rule 26 25.11.2025] In some embodiments, the infectious disease described above comprises one or more of infections by viruses, fungi, and bacteria.

[0499] [Corrected according to Rule 26 25.11.2025] In some embodiments, the autoimmune disease described above comprises one or more of: acute idiopathic thrombocytopenic purpura, chronic idiopathic thrombocytopenic purpura, systemic lupus erythematosus, rheumatoid arthritis, psoriasis, inflammatory bowel disease, multiple sclerosis, celiac disease, type 1 diabetes, and diffuse toxic goiter.

[0500] [Corrected according to Rule 26 25.11.2025] In some embodiments, the genetic disease described above comprises one or more of: hemophilia, thalassemia, and Gaucher’s disease.

[0501] [Corrected according to Rule 26 25.11.2025] In some embodiments, the neurological disease described above comprises one or more of: amyotrophic lateral sclerosis, Alzheimer’s disease, and glioma.

[0502] [Corrected according to Rule 26 25.11.2025] In some embodiments, the drug described above is a vaccine.

[0503] [Corrected according to Rule 26 25.11.2025] In some embodiments, the drug described above is a nucleic acid drug, wherein the nucleic acid comprises at least one of: RNA and DNA. In an alternative specific example, the DNA is a plasmid. In an alternative specific example, the RNA is one or more of: mRNA, antisense oligonucleotide, rRNA, miRNA, tRNA, siRNA, snRNA, shRNA, sgRNA, and Cas9 mRNA.

[0504] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure provides use of the lipid nanoparticle of any of the embodiments described above in the manufacture of a carrier for the transfer or delivery of an active ingredient. In some embodiments, the active ingredient is a pharmaceutically active ingredient, such as a nucleic acid.

[0505] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a method of delivering the active ingredient described above to a mammalian cell, the method comprising: administering to a mammal the lipid nanoparticle comprising the active ingredient in any of the embodiments described above, the administering comprising contacting a cell of the mammal with the lipid nanoparticle, thereby delivering the active ingredient to the cell of the mammal. In some embodiments, the active ingredient is a pharmaceutically active ingredient, e.g., a nucleic acid.

[0506] [Corrected according to Rule 26 25.11.2025] In some embodiments, the nucleic acid comprises at least one of: RNA, messenger RNA (mRNA), antisense oligonucleotide, DNA, plasmid, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA), and Cas9 mRNA.

[0507] [Corrected according to Rule 26 25.11.2025] In other embodiments, the active ingredient is a transfection reagent or a detection reagent.

[0508] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a medicament comprising the PEG lipid of any of the embodiments described above, the lipid nanoparticle of any of the embodiments described above, or the pharmaceutical composition of any of the embodiments described above.

[0509] [Corrected according to Rule 26 25.11.2025] In some embodiments, the disease or condition that can be prevented or treated with the medicament is as described above.

[0510] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a medicament comprising the medicament made from the pharmaceutical composition of any of the embodiments described above.

[0511] [Corrected according to Rule 26 25.11.2025] In some embodiments, the disease or condition that can be prevented or treated with the medicament is as described above.

[0512] [Corrected according to Rule 26 25.11.2025] In addition, the present disclosure also provides a method of preventing or treating a disease or condition, comprising the step of administering to a subject in need thereof the lipid nanoparticle, the pharmaceutical composition, or the medicament of the present disclosure.

[0513] [Corrected according to Rule 26 25.11.2025] In some embodiments, the disease or condition that is prevented or treated is as described above.

[0514] [Amended according to Rule 26 on 25.11.2025] The above-mentioned methods for preventing or treating diseases have their corresponding advantages due to the use of the above-mentioned lipid nanoparticles, pharmaceutical compositions or drugs.

[0515] [Amended according to Rule 26 on 25.11.2025] In addition, the present disclosure also provides a method for reducing accelerated blood clearance (ABC) in a subject receiving treatment, the method comprising delivering a pharmaceutically active ingredient using a lipid nanoparticle (LNPs) containing the PEG lipid of any of the above-mentioned embodiments.

[0516] [Amended according to Rule 26 on 25.11.2025] In addition, the present disclosure also provides a method for delivering lipid nanoparticles (LNPs) encapsulating a pharmaceutically active ingredient to a subject without affecting the immune response promoting accelerated blood clearance (ABC) of subsequent doses of LNPs, the method comprising:

[0517] [Amended according to Rule 26 on 25.11.2025] The LNPs encapsulating a pharmaceutically active ingredient are provided, which comprise the PEG lipid of any of the above-mentioned embodiments;

[0518] [Amended according to Rule 26 on 25.11.2025] A first dose of LNPs is administered to a subject, wherein the first dose of LNPs does not induce an immune response promoting ABC when a second dose of LNPs is administered; and

[0519] [Amended according to Rule 26 on 25.11.2025] A second dose of LNPs is administered to a subject, wherein the subject does not have an ABC response to the second dose of LNPs. [Amended according to Rule 26 on 25.11.2025] Embodiments

[0520] [Amended according to Rule 26 on 25.11.2025] In order to make the purposes and technical solutions of the present disclosure clearer, the following will be described in detail in combination with specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present disclosure. In the embodiments, reagents and instruments are used as non-specifically described, which are conventional choices in the art. The experimental methods not specified in the embodiments are implemented according to conventional conditions, such as conditions described in the literature, books or methods recommended by manufacturers.

[0521] [Amended according to Rule 26 on 25.11.2025] In the examples, "Hep" refers to heptane, "EA" refers to ethyl acetate, "DMF" refers to N,N-dimethylformamide, "TEA" refers to triethylamine, "EDCI" refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, "DMAP" refers to 4-dimethylaminopyridine, and "DCM" refers to dichloromethane.

[0522] [Amended according to Rule 26 on 25.11.2025] Synthesis process of compound 001 of Example 1

[0523] [Amended according to Rule 26 on 25.11.2025] First step Synthesis of compound 001-A1

[0524] [Amended according to Rule 26 on 25.11.2025] Experimental process

[0525] [Amended according to Rule 26 on 25.11.2025] Into a 250 mL single-necked flask, 1-hexadecanol (10.0 g, 41.25 mmol) was dissolved in DMF (100 mL) and cooled to below 10 °C. NaH (4.95 g, 123.75 mmol) was slowly added in portions, and after the addition was completed, the mixture was stirred for 30 min. Bromoacetic acid (6.88 g, 49.5 mmol) was slowly added to the reaction, and after the addition was completed, the reaction was continued for 30 min, and then the temperature was raised to room temperature (25 °C) and the reaction was continued for 15 h. Water (200 mL) was added to the reaction, the pH was adjusted to 6.5-7.5 with dilute hydrochloric acid, and then EA (200 mL) was added twice, the EA combined phase was washed with saturated brine (200 mL), and the EA phase was collected and distilled under reduced pressure. The concentrate was subjected to silica gel column chromatography and eluted with Hep:EA = 3:1 to obtain compound 001-A1 as a white solid, 4.5 g, in a yield of 36.3%.

[0526] [Amended according to Rule 26 on 25.11.2025] Second step Synthesis of compound 001

[0527] [Amended according to Rule 26 on 25.11.2025] Experimental process

[0528] [Amended according to Rule 26 on 25.11.2025] Into a 250 mL single-necked flask, PEG2000 (19.96 g, 9.98 mmol) was added, DCM (100 mL) was added, and it was dissolved under ultrasonic, then EDCI (1.91 g, 9.98 mmol), TEA (1.05 g, 9.98 mmol), DMAP (0.24 g, 2.0 mmol) were added, and after stirring for 5 min, compound 001-A1 (2 g, 6.66 mmol) was added, and the reaction was carried out at room temperature for 15 h. Water (200 mL) was added to the reaction, stirred for 10 min, and the lower organic phase was collected, and the aqueous phase was extracted with DCM (100 mL) once, and the combined DCM phase was washed with saturated brine (100 mL) once, and finally the DCM phase was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography, eluted with Hep:EA = 15:1, and the obtained product was further purified by reverse phase column chromatography, eluted with EtOH:H2O gradient (20%~70%), to obtain compound 001 (800 mg), purity 99.3%, yield 5.3%.

[0529] [Amended according to Rule 26 on 25.11.2025] 1 H NMR (400 MHz, CDCl3) δ 4.31 (m, 2H), 4.10 (s, 2H), 3.76-3.57 (m, 184H), 3.52 (t, J = 6.7 Hz, 2H), 1.67-1.55 (m, 2H), 1.40-1.20 (m, 26H), 0.88 (t, J = 6.8 Hz, 3H).

[0530] [Amended according to Rule 26 on 25.11.2025] LC-MS (ESI): Mn = 2277.

[0531] [Amended according to Rule 26 on 25.11.2025] Synthesis process of compound 003 of Example 2

[0532] [Amended according to Rule 26 on 25.11.2025] First step Synthesis of compound 003-A1

[0533] [Amended according to Rule 26 on 25.11.2025] Experimental process

[0534] [Amended according to Rule 26 on 25.11.2025] Into a 250 mL single-necked flask, 1-octadecylamine (5 g, 18.55 mmol) was added, and then it was dissolved in DCM (100 mL), and then saturated aqueous sodium bicarbonate solution (50 mL) was added, and the temperature was lowered to below 10 °C, and then triphosgene (2.2 g, 7.42 mmol) was added to the reaction, and the reaction was carried out for 1 h, and then the organic phase was collected by separation, and the aqueous phase was extracted once more with DCM (100 mL), and then the DCM phase was washed once with saturated brine (100 mL), and then the DCM phase was concentrated under reduced pressure. The concentrate was subjected to silica gel column chromatography, and then eluted with Hep:EA = 5:1 to obtain compound 003-A1 as a solid 4.64 g, with a yield of 84.6%.

[0535] [Amended according to Rule 26 on 25.11.2025] Synthesis of compound 003

[0536] [Amended according to Rule 26 on 25.11.2025] Experimental process

[0537] [Amended according to Rule 26 on 25.11.2025] Into a 250 mL single-necked flask, PEG2000 (30.46 g, 15.23 mmol) was added, and then it was dissolved in DMF (60 mL), and then cuprous iodide (0.39 g, 2.03 mmol) was added, and then compound 003-A1 (3 g, 10.15 mmol) was added, and the reaction was carried out at room temperature for 3 h. The solid was removed by filtration, and then water (200 mL) was added to the filtrate, and then it was extracted twice with EA (100 mL), and then ammonia water was added dropwise under stirring to remove copper ions until the EA phase was colorless. The EA phases were combined, and then the EA phase was washed once more with saturated brine (100 mL), and then the EA phase was concentrated under reduced pressure. The concentrate was subjected to silica gel column chromatography, and then eluted with Hep:EA = 5:1, and then the obtained product was purified again using reverse phase column chromatography, and then eluted with an EtOH:H2O gradient (20% to 70%) to obtain compound 003 (1.4 g) with a purity of 98.9% and a yield of 6.2%.

[0538] [Amended according to Rule 26 on 25.11.2025] 1 H NMR (400 MHz, CDCl3) δ 4.25-4.16 (m, 2H), 3.86-3.78 (m, 1H), 3.77-3.53 (m, 184H), 3.15 (dd, J = 13.4, 6.8 Hz, 2H), 1.47 (m, 2H), 1.25 (m, 30H), 0.88 (t, J = 6.8 Hz, 3H).

[0539] [Amended according to Rule 26 on 25.11.2025] LC-MS (ESI): Mn = 2230.

[0540] [Corrected according to Rule 26 on 25.11.2025] Synthesis process of compound 006 of Example 3

[0541] [Corrected according to Rule 26 on 25.11.2025] First step Synthesis of compound 006-A1

[0542] [Corrected according to Rule 26 on 25.11.2025] Experimental process

[0543] [Corrected according to Rule 26 on 25.11.2025] Into a 100 mL single-necked flask, 1-hexadecanol (5 g, 20.62 mmol) was dissolved in DCM (50 mL), pyridine (2.45 g, 30.39 mmol), DMAP (0.76 g, 6.19 mmol) were added, and the mixture was stirred at room temperature. Then p-nitrophenyl chloroformate (4.99 g, 24.74 mmol) was added portionwise slowly. The mixture was stirred at room temperature for 3 h. Water (100 mL) was added, and the organic phase was collected by stirring and liquid-liquid separation. The aqueous phase was extracted with DCM (100 mL), and the combined DCM phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The concentrate was purified by column chromatography on silica gel with Hep:EA = 20:1 as the eluent to obtain compound 006-A1, a solid, 8.1 g, in a yield of 96.9%.

[0544] [Corrected according to Rule 26 on 25.11.2025] Second step Synthesis of compound 006

[0545] [Corrected according to Rule 26 on 25.11.2025] Experimental process

[0546] [Corrected according to Rule 26 on 25.11.2025] Into a 500 mL single-necked flask, PEG2000 (49.16 g, 24.58 mmol) was dissolved in DCM (150 mL), pyridine (4.9 g, 61.45 mmol), DMAP (0.45 g, 3.69 mmol) were added, and finally compound 006-A1 (5 g, 12.29 mmol) was added. The mixture was stirred at room temperature for 3 days. Water (200 mL) was added to the reaction solution, and the organic phase was collected by stirring and liquid-liquid separation. The aqueous phase was extracted with DCM (100 mL), and the combined organic phases were washed with saturated brine (100 mL) once, and the organic phase was collected and concentrated under reduced pressure. The concentrate was purified by column chromatography on silica gel with Hep:EA = 15:1 as the eluent. The obtained product was further purified by column chromatography on a reversed phase with EtOH:H2O gradient elution (20% to 70%) to obtain compound 006 (9.3 g) with a purity of 99.1% and a yield of 31.7%.

[0547] [Corrected according to Rule 26 on 25.11.2025] 1H NMR (400 MHz, CDC13) δ 4.27 (m, 2H), 4.12 (t, J = 6.8 Hz, 2H), 3.65 (d, J = 7.7 Hz, 184H), 1.70 - 1.59 (m, 2H), 1.38 - 1.21 (m, 30H), 0.88 (t, J = 6.8 Hz, 3H).

[0548] [Amended according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2156.

[0549] [Amended according to Rule 26 25.11.2025] Synthesis of compound 002 of Example 4

[0550] [Amended according to Rule 26 25.11.2025] Synthesis of compound 002-A1 by reference to the first step of Example 3, then synthesis of compound 002 using compound 002-A1 and by reference to the second step of Example 3, wherein compound 002-A1 (2 g, 4.59 mmol), PEG2000 (13.8 g, 6.89 mmol) gave compound 002 (1.67 g) with purity 98.3% and yield 15.6%.

[0551] [Amended according to Rule 26 25.11.2025] 1 H NMR (400 MHz, CDC13) δ 4.27 (m, 2H), 4.12 (t, J = 6.8 Hz, 2H), 3.65 (d, J = 7.7 Hz, 184H), 1.70 - 1.59 (m, 2H), 1.38 - 1.21 (m, 30H), 0.88 (t, J = 6.8 Hz, 3H).

[0552] [Amended according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2240

[0553] [Amended according to Rule 26 25.11.2025] Synthesis of compound 004 of Example 5

[0554] [Amended according to Rule 26 25.11.2025] Synthesis of compound 004-A1 by reference to the first step of Example 3, then synthesis of compound 004 using compound 004-A1 and by reference to the second step of Example 3, wherein compound 004-A1 (1.5 g, 3.34 mmol), PEG2000 (10.0 g, 5.00 mmol) gave compound 004 (2.3 g) with purity 98.8% and yield 30.14%.

[0555] [Corrected according to Rule 26 25.11.2025] 1 H NMR (400 MHz, CDC13) δ 4.27 (m, 2H), 4.12 (t, J = 6.8 Hz, 2H), 3.65 (m, 184H), 1.63 (m, 2H), 1.28 (m, 34H), 0.88 (t, J = 6.8 Hz, 3H).

[0556] [Corrected according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2243.

[0557] [Corrected according to Rule 26 25.11.2025] Synthesis of compound 005 of example 6

[0558] [Corrected according to Rule 26 25.11.2025] Compound 005-A1 was synthesized by referring to the first step of example 3, then compound 005 was synthesized by referring to the second step of example 3 using compound 005-A1 (1.2 g, 2.59 mmol), PEG2000 (7.76 g, 3.88 mmol), compound 005 (2.12 g) was obtained with purity 98.1% and yield 36.49%.

[0559] [Corrected according to Rule 26 25.11.2025] 1 H NMR (400 MHz, CDC13) δ 4.27 (m, 2H), 4.12 (t, J = 6.8 Hz, 2H), 3.65 (m, 184H), 1.63 (m, 2H), 1.28 (m, 34H), 0.88 (t, J = 6.8 Hz, 3H).

[0560] [Corrected according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2243.

[0561] [Corrected according to Rule 26 25.11.2025] Synthesis of compound 013 of example 7

[0562] [Corrected according to Rule 26 25.11.2025] Compound 013-A1 was synthesized by referring to the first step of example 3, then compound 013 was synthesized by referring to the second step of example 3 using compound 013-A1 (1.6 g, 3.35 mmol), PEG2000 (10.05 g, 5.02 mmol), compound 013 (2.9 g) was obtained with purity 98.5% and yield 39.7%.

[0563] [Rule 26 Correction 25.11.2025] 1 H NMR (400 MHz, CDC13) δ 4.31 4.25 (m, 2H), 4.12 (t, J = 6.8 Hz, 2H), 3.65 (m, 184H), 1.63 (m, 2H), 1.25 (m, 36H), 0.88 (t, J = 6.8 Hz, 3H).

[0564] [Rule 26 Correction 25.11.2025] LC-MS (ESI): Mn = 2180.

[0565] [Rule 26 Correction 25.11.2025] Synthesis of compound 014

[0566] [Rule 26 Correction 25.11.2025] Compound 014-A1 was synthesized by following the first step of Reference Example 3, then compound 014 was synthesized by using compound 014-A1 and following the procedure of the second step of Reference Example 3, wherein compound 014-A1 (2 g, 4.07 mmol), PEG2000 (12.2 g, 6.11 mmol) to give compound 014 (2.58 g) with purity 99.2% and yield 29.5%.

[0567] [Rule 26 Correction 25.11.2025] 1 H NMR (600 MHz, CDC13) δ 4.26-4.19 (m, 2H), 4.15-4.09 (t, J = 6.8 Hz, 2H), 3.77-3.42 (m, 184H), 1.65-1.55 (m, 2H), 1.34-1.15 (m, 38H), 0.83 (t, J = 6.8 Hz, 3H).

[0568] [Rule 26 Correction 25.11.2025] LC-MS (ESI): Mn = 2150.

[0569] [Rule 26 Correction 25.11.2025] Synthesis of compound 015

[0570] [Rule 26 Correction 25.11.2025] Compound 015-A1 was synthesized by following the first step of Reference Example 3, then compound 015 was synthesized by using compound 015-A1 and following the procedure of the second step of Reference Example 3, wherein compound 015-A1 (2.1 g, 5.98 mmol), PEG2000 (17.94 g, 8.96 mmol) to give compound 015 (2.68 g) with purity 98.2% and yield 20.1%.

[0571] [Corrected according to Rule 26 25.11.2025] 1 H NMR (600 MHz, CDC13) δ 4.33 - 4.26 (m, 2H), 4.14 (t, J = 6.8 Hz, 2H), 3.87 - 3.58 (m, 184H), 1.72 - 1.62 (m, 2H), 1.41 - 1.24 (m, 18H), 0.90 (t, J = 6.8 Hz, 3H).

[0572] [Corrected according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2230.

[0573] [Corrected according to Rule 26 25.11.2025] Synthesis of compound 016 of Example 10

[0574] [Corrected according to Rule 26 25.11.2025] Compound 016-A1 was synthesized by following the first step of Reference Example 3, then compound 016-A1 (2.3 g, 6.06 mmol) was used and compound 016 was synthesized by following the second step of Reference Example 3, wherein compound 016-A1 (2.3 g, 6.06 mmol), PEG2000 (18.2 g, 9.09 mmol) to give compound 016 (3.0 g) with purity 98.9% and yield 22.1%.

[0575] [Corrected according to Rule 26 25.11.2025] 1 H NMR (600 MHz, CDC13) δ 4.33 - 4.26 (m, 2H), 4.14 (t, J = 6.8 Hz, 2H), 3.87 - 3.58 (m, 184H), 1.72 - 1.62 (m, 2H), 1.41 - 1.24 (m, 18H), 0.90 (t, J = 6.8 Hz, 3H).

[0576] [Corrected according to Rule 26 25.11.2025] LC-MS (ESI): Mn = 2240.

[0577] [Corrected according to Rule 26 25.11.2025] Synthesis of compound 007 of Example 11

[0578] [Amended according to Rule 26 on 25.11.2025] The compound 007-A1 was synthesized according to the first step of Example 3, then the compound 007-A1 was used and the compound 007 was synthesized according to the second step of Example 3, wherein compound 007-A1 (1.5 g, 3.56 mmol), PEG2000 (28.48 g, 14.24 mmol), to obtain compound 007 (2.61 g), purity 99.1%, yield 33.2%.

[0579] [Amended according to Rule 26 on 25.11.2025] 1 H NMR (600 MHz, CDCl3) δ 4.29-4.23 (t, J = 4.8 Hz, 2H), 4.13-4.07 (t, J = 6.8 Hz, 2H), 3.62 (m, 184H), 1.65-1.59 (m, 2H), 1.35-1.29 (m, 2H), 1.29-1.19 (m, 28H), 0.87 (t, J = 6.8 Hz, 3H).

[0580] [Amended according to Rule 26 on 25.11.2025] LC-MS (ESI): Mn = 2211.

[0581] [Amended according to Rule 26 on 25.11.2025] Test Example 1. Preparation of lipid nanoparticles encapsulating human erythropoietin mRNA (hEPO mRNA)

[0582] [Amended according to Rule 26 on 25.11.2025] (1) Ingredients:

[0583] [Amended according to Rule 26 on 25.11.2025] The specified amount of hEPO mRNA stock solution, 0.2M sodium acetate buffer, DEPC water were added to the container, mixed to prepare the aqueous phase.

[0584] [Amended according to Rule 26 on 25.11.2025] The cationic lipid (XI-1), the helper lipid (DSPC), the structural lipid (cholesterol), and the PEG lipid of the present disclosure or the control PEG lipid (DMG-PEG2000) were respectively dissolved in anhydrous ethanol to prepare respective solutions, with concentrations of 20 mg / mL, 10 mg / mL, 20 mg / mL and 25 mg / mL, respectively. According to the molar ratio of cationic lipid (XI-1): DSPC: CHO-HP: PEG lipid is 48:10:40.5:1.5, the above four solutions were respectively removed and mixed to prepare the alcohol phase.

[0585] [Corrected according to Rule 26 25.11.2025] (2) Encapsulation: using a microfluidic device (MPE-L2) to suck the aqueous phase and the alcohol phase into the aqueous phase and alcohol phase syringes at flow rates of 9 mL / min: 3 mL / min, respectively, and to encapsulate at a flow rate of aqueous phase: alcohol phase = 12 mL / min: 4 mL / min to obtain mRNA-encapsulated lipid nanoparticles (mRNA-LNP).

[0586] [Corrected according to Rule 26 25.11.2025] (3) Dialysis: load the product of step (2) into a dialysis bag and place it in a Tris Buffer-8% (m / V) sucrose solution for replacement to remove residual ethanol, un-assembled lipids, and other components. Dialyze for 2 hours at room temperature in the dark with magnetic stirring (replace the dialysate every 1 hour).

[0587] [Corrected according to Rule 26 25.11.2025] (4) Pass the product of step (3) through a 0.22 μm microporous filter to sterilize, then aliquot to prepare a plurality of hEPO mRNA-encapsulated lipid nanoparticle preparations (LNP preparations), wherein the PEG lipid contained in each LNP preparation is different, the concentration of hEPO mRNA in each LNP preparation is 0.2 μg / μL, the molar ratio of hEPO mRNA: Lipid (cationic lipid (XI-1)) is 1:4, the particle size is 80 nm-130 nm, and the encapsulation efficiency is more than 85%.

[0588] [Corrected according to Rule 26 25.11.2025] Test Example 2. Detection of hEPO concentration in serum of rats after multiple dosing

[0589] [Corrected according to Rule 26 25.11.2025] Animal preparation: select 5-7 week old SD rats, half male and half female, and raise them in a SPF level feeding room. Animal testing is strictly in accordance with the guidelines of the national health agency and the requirements of animal ethics.

[0590] [Corrected according to Rule 26 25.11.2025] In vivo delivery: before injection of the test LNP preparation, gently invert the LNP preparation repeatedly to mix the preparation sample completely. Use a 2 mL syringe to draw the corresponding amount of LNP preparation, and inject the LNP preparation by tail vein injection (IV), repeat 4-6 rats for each LNP preparation, half male and half female, and inject each rat with the corresponding LNP preparation encapsulating human erythropoietin mRNA (hEPO mRNA) prepared in Test Example 1 at a dose of 0.5 mg / kg mRNA; administer once a week for 4-6 times.

[0591] [According to the rules 26 correction 25.11.2025] The whole blood of rats was collected after the first injection of LNP preparation 3h / 8h / 24h, and the serum was obtained after the second, third, fourth, fifth and sixth injection of LNP preparation 8h, respectively. Among them, the steps of obtaining serum include: collecting rat blood samples and placing them in test tubes without anticoagulant; after the blood coagulates at room temperature for 30-60 min, centrifuge at 3500 rpm for 10 min to obtain the supernatant, which is the serum.

[0592] [According to the rules 26 correction 25.11.2025] The specific method for detecting hEPO concentration in serum is as follows:

[0593] [According to the rules 26 correction 25.11.2025] 1. Solution preparation

[0594] [According to the rules 26 correction 25.11.2025] 0.05% PBST washing solution: take 250 mL 20x PBS, add water to 5 L, add 2.5 mL Tween20 and mix well for standby; 3% BSA blocking solution: electronic balance 3.0g BSA powder, add 0.05% PBST to 100mL and mix well for standby; 1% BSA dilution: electronic balance 1.0g BSA powder, add 0.05% PBST to 100mL and mix well for standby; standard curve reagent: 1) stock solution dilution: take stock solution (item number: EPO-H4214 manufacturer: Acro) 2μL, add 998μL diluent and mix well, the concentration of the standard is 1.2μg / mL; 2) standard curve dilution: 3-fold gradient dilution.

[0595] [According to the rules 26 correction 25.11.2025] 2. ELISA detection:

[0596] [Rule 26 Correction 25.11.2025] (1) Coating: dilute the coating antibody (item number: ab272358, manufacturer: Abeam) to 1 pg / mL with PBS, 100 pL / well, coat overnight at 4°C; (2) Wash plate: wash the plate with 0.05% PBST wash solution, 250 pL / well, wash 3 times; (3) Blocking: prepare 3% BSA with 0.05% PBST wash solution, 250 pL / well, incubate at 37°C for 1 h; (4) Wash plate: wash the plate with 0.05% PBST wash solution, 250 pL / well, wash 3 times; (5) Incubate the sample to be tested: add the diluted standard curve and test sample according to the plate layout, 100 pL / well, incubate at 37°C for 2 h; (6) Wash plate: wash the plate with 0.05% PBST wash solution, 250 pL / well, wash 3 times; (7) Incubate the detector antibody: prepare 0.2 pg / mL detector antibody (biotin-labeled) with 1% BSA diluent, 100 pL / well, incubate at 37°C for 1 h; (item number: ab272359, manufacturer: Abeam); (8) Wash plate: wash the plate with 0.05% PBST wash solution, 250 pL / well, wash 3 times; (9) SA-HRP incubation: dilute the detection secondary antibody (item number: 21126, manufacturer: Invitrogen) with diluent 1:5000, 100 pL / well, incubate at 37°C for 45 min; (10) Wash plate: wash the plate with 0.05% PBST diluent, 250 pL / well, wash 3 times; (11) Color development: add TMB (item number: PR1200, manufacturer: Solarbio) for color development, 100 pL / well, incubate at 37°C for 5 min; (12) Stop: add stop solution, 50 pL / well; (13) Read plate: enzyme label OD450 and OD630; (14) Data analysis: OD450-OD630.

[0597] [Rule 26 Correction 25.11.2025] The results of the detection of hEPO concentration in serum are shown in Figure 1. As can be seen from Figure 1, when multiple doses are administered, the serum hEPO concentration of rats administered with LNP formulations containing DMG-PEG2000 gradually decreases with the increase in the number of doses, which suggests that the LNP formulation containing DMG-PEG2000 has an accelerated blood clearance (ABC) phenomenon. When multiple doses are administered, the serum hEPO concentration of rats administered with LNP formulations containing the PEG lipid of the present disclosure does not gradually decrease with the increase in the number of doses, indicating that the LNP formulation containing the PEG lipid of the present disclosure can significantly alleviate the accelerated blood clearance (ABC) phenomenon.

[0598] [Rule 26 Correction 25.11.2025] Test Example 3. Detection of anti-PEG IgG in serum of rats after multiple doses

[0599] [Corrected according to Rule 26 25.11.2025] Serum collection: The rats in Test Example 2 were collected whole blood at 96h or 120h after each administration, and serum was obtained, the method of obtaining serum is shown in Test Example 2.

[0600] [Corrected according to Rule 26 25.11.2025] Serum Anti-PEG IgG detection specific method as follows:

[0601] [Corrected according to Rule 26 25.11.2025] 1. Solution preparation

[0602] [Corrected according to Rule 26 25.11.2025] 0.05% CHAPS washing solution: take 200mL 20xPBS, add water to constant volume to 4L, add 2g CHAPS, mix well, ready for use;

[0603] [Corrected according to Rule 26 25.11.2025] 2.3% BSA blocking solution: weigh 9.0g BSA powder, add 0.05% PBS-CHAPS to constant volume to 300mL, mix well for standby; 3.1% BSA diluent: weigh 1.0g BSA powder, add 0.05% PBS-CHAPS to constant volume to 100mL, mix well for standby;

[0604] [Corrected according to Rule 26 25.11.2025] 2. ELISA detection

[0605] [Amended according to Rule 26 on 25.11.2025](1) Coating reagent: 0.1M sodium carbonate-bicarbonate solution (pH = 9.51) Dilute the coating reagent (coating the corresponding PEG) to 10 pg / mL, 100 pL / well, 4°C overnight; (2) Washing plate: use 0.05% CHAPS wash solution (item number: ST1146-5g, manufacturer: Biyun Tian) to wash the plate, 250 pL / well, wash 3 times; (3) Blocking: use 0.05% CHAPS wash solution with 3% BSA, 250 pL / well, 37°C incubate for 1.5h; (4) Washing plate: use 0.05% CHAPS wash solution to wash the plate, 250 pL / well, wash 3 times; (5) Incubate the sample to be tested: add the diluted test sample according to the plate layout, 100 pL / well, 37°C incubate for 2h; (6) Washing plate: use 0.05% CHAPS wash solution to wash the plate, 250 pL / well, wash 3 times; (7) Rat-HRP incubation: dilute the detection secondary antibody (item number: 62-9520, manufacturer: Invitrogen) to 1:4000 in 0.05% CHAPS wash solution with 1% BSA, 100 pL / well, 37°C incubate for 1h; (8) Washing plate: use 0.05% PBST diluent to wash the plate, 250 pL / well, wash 3 times; (9) Color development: add TMB color developing agent, 100 pL / well, 37°C incubate for 10min; (10) Termination: add termination solution, 50 pL / well; (11) Plate reading: enzyme marker OD450 and OD630; (12) Data analysis: OD450-OD630.

[0606] [Amended according to Rule 26 on 25.11.2025] The detection results of anti-PEG IgG in the serum of rats 96h after the third administration are shown in Figure 2. As can be seen from Figure 2, the anti-PEG IgG antibody titer in the serum of rats administered with LNP formulations containing PEG lipids of the present disclosure is extremely low, especially the LNP formulations containing 001, the LNP formulations containing 002, the LNP formulations containing 006 and the LNP formulations containing 007, the detection results of anti-PEG IgG antibodies in the serum of rats are basically negative, which indicates that LNPs containing PEG lipids of the present disclosure can significantly reduce the production of anti-PEG IgG antibodies or do not produce anti-PEG IgG antibodies.

[0607] [Amended according to Rule 26 on 25.11.2025] Detection of cytokines IL-6 and IFN-γ in the serum of rats after administration in Test Example 4

[0608] [Amended according to Rule 26 on 25.11.2025] Serum acquisition: rats in Test Example 2, collect whole blood 5h after the first administration, obtain serum, use kit (LEGENDplex TMRat Inflammation Panel (13-plex) w / VbP V02 (BIOLEGEND, cat# 741396) was used to detect the content of IL-6 and IFN-γ in serum, and the specific steps included:

[0609] [According to Rule 26 Correction 25.11.2025] 1. Prepare 1x Wash Buffer, standards and Beads according to the method provided in the kit.

[0610] [According to Rule 26 Correction 25.11.2025] 2. Sample addition and incubation: Add standards and 4-fold diluted rat serum samples to the V-bottom Plate, respectively, then add Beads to each well and shake the V-bottom Plate on the microplate shaker at 500 rpm, incubate at room temperature for 2 h in the dark; after incubation, wash the plate with 1x Wash Buffer.

[0611] [According to Rule 26 Correction 25.11.2025] 3. Detection antibody addition: Add 25 μL of Detection Antibodies to each well of the V-bottom Plate, shake the V-bottom Plate on the microplate shaker at 500 rpm, incubate at room temperature for 1 h in the dark.

[0612] [According to Rule 26 Correction 25.11.2025] 4. After incubation, directly add 25 μL of SA-PE to each well, shake the V-bottom Plate on the microplate shaker at 500 rpm, incubate at room temperature for 30 min in the dark.

[0613] [According to Rule 26 Correction 25.11.2025] 5. After incubation, wash the plate once, then add 200 μL of 1x Wash Buffer to resuspend, and detect by flow cytometry.

[0614] [According to Rule 26 Correction 25.11.2025] The results are shown in Figures 3 and 4. There are obvious differences in the cytokine responses induced by LNP formulations prepared from PEG lipids of different structures. When the linking groups connecting the polyethylene glycol moiety and the alkyl tail chain in the PEG lipid are different, the content of IL-6 or IFN-γ induced by the LNP formulations prepared from the corresponding PEG lipids is significantly different. For example, the content of IFN-γ induced by the LNP formulation prepared from the PEG lipid shown in 006 is significantly lower than that induced by the LNP formulation prepared from the PEG lipid shown in 001.

Claims

1. [Corrected pursuant to Rule 26 25.11.2025] A PEG lipid as shown in formula (I): or a salt thereof, or a stereoisomer thereof, wherein R1is H, HO-Z1-Q1-, or H-Z1-Q1-, Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond, Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond, Q1is -C(=O)-, -OC(=O)-, or a bond, Q2is -C(=O)-, or a bond, Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond, L1is C1to C 20 alkylene or a bond, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond, R2is C4to C 24 hydrocarbyl, the total number of carbon atoms of L1and R2is 10 to 35, L2is C1to C4alkylene, L3is C1to C4alkylene, R3is H or C1to C3alkyl, n is any integer from 30 to 60.

2. [Corrected pursuant to Rule 26 25.11.2025] A PEG lipid as shown in formula (IX): or a salt thereof, or a stereoisomer thereof, wherein R1is H, HO-Z1-Q1-, or H-Z1-Q1-, Y is -C(=O)-, -C(=O)O-, -C(=O)NH-, -Q2-Z2-Q3-, or a bond, Z1and Z2are each independently -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond, Q1is -C(=O)-, -OC(=O)-, or a bond, Q2is -C(=O)-, or a bond, Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond, L1is C1to C 20 alkylene, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond, R2is C4to C 24 hydrocarbyl, the total number of carbon atoms of L1and R2is 10 to 35, L2is C1to C4alkylene, L3is C1to C4alkylene, R3is H or C1to C3alkyl, n is any integer from 30 to 60. L1and A1and R2form a four-, five-, or six-membered ring.

3. A PEG lipid as shown in formula (X): ###0002### (X) [Corrected according to Rule 26 25.11.2025] or a salt thereof, or a stereoisomer thereof, wherein R1is H, HO-Z1-Q1-, or H-Z1-Q1-, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, -L2-, or a bond, Z2is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3-, or -L2-, Q1is -C(=O)-, -OC(=O)-, or a bond, Q2is -C(=O)-, or a bond, Q3is -C(=O)O-, -OC(=O)-, -O-, -S-, or a bond, L1is C1to C 20 alkylene or a bond, A1is -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -S-, or a bond, R2is C4to C 24 hydrocarbyl, the total number of carbon atoms of L1and R2is 10 to 35, L2is C1to C4alkylene, L3is C1to C4alkylene, R3is H or C1to C3alkyl, n is any integer from 30 to 60. Z2forms a four-, five- or six-membered ring with Q3, L1, A1and R2.

4. [Amended according to Rule 26 25.11.2025] The PEG lipid according to any one of claims 1 to 3, wherein R1is H, Preferably, R1 is H, 5. The PEG lipid of claim 1 or 4, wherein the structure of the PEG lipid is according to Formula (II): ###0002### (II) [Amended according to Rule 26 25.11.2025] wherein R1is not H and not unsubstituted methyl.

6. [Amended according to Rule 26 25.11.2025] The PEG lipid according to claim 5, wherein (1) R1is HO-Z1-Q1-, A1is a bond, and Z1and Q1are not simultaneously a bond; Preferably, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3- or -L2-; More preferably, Z1is -L2-CH(OR3)-L3-, -L2-C(=O)-L3-, -L2-CH(R3)-L3- or -L2-, and Q1is -C(=O)-, -OC(=O)- or a bond, wherein L2is preferably C1-C3alkylene, L3is preferably C1-C3alkylene, and R3is preferably H or C1-C3alkyl; Preferably, R1 is or (2) R1is H-Z1-Q1-, wherein Z1is not a bond and not unsubstituted methylene when Q1is a bond; Preferably, R1is H-Z1-Q1-, Z1is -L2-, Q1is -C(=O)-, and A1is a bond, wherein L2is preferably C1-C3alkylene, more preferably C1-C3alkanediyl, More preferably, R1is 7. The PEG lipid of claim 1, wherein the PEG lipid is according to Formula (II-1): ###0002### (II-1) [Amended according to Rule 26 25.11.2025] wherein A1is -OC(=O)-, -OC(=O)O-, -O- or -S-; Preferably, the PEG lipid is of formula (II-1-1):

8. The PEG lipid of claim 1 or 4, wherein the PEG lipid has a structure as shown in Formula (III), Formula (IV), Formula (V), Formula (VI), or Formula (VII):

9. The PEG lipid of any one of claims 1 and 4-8, wherein the PEG lipid has the structure: [RULE 26] 25.11.2025 wherein in said formula (II-2-3), formula (II-2-4), formula (VII-1) or formula (VII-2), R3is preferably H or C1-C3alkyl, more preferably H, C1alkyl or C2alkyl.

10. The PEG lipid of claim 1 or 4, wherein the PEG lipid has the structure:

11. The PEG lipid of claim 10, wherein the PEG lipid has the structure: [Amended under Rule 26 25.11.2025] 12. [Amended according to Rule 26 25.11.2025] The PEG lipid according to any one of claims 1 to 11, wherein the total number of carbon atoms of L1and R2is 12 to 25.

13. [Amended according to Rule 26 25.11.2025] The PEG lipid according to claim 12, wherein the total number of carbon atoms of L1and R2is 16 to 22.

14. [Amended according to Rule 26 25.11.2025] The PEG lipid according to any one of claims 1 to 13, wherein L1is C1-C8alkylene or a bond.

15. [Amended according to Rule 26 25.11.2025] The PEG lipid according to claim 14, wherein L1is C1-C8alkanediyl, C2-C8alkenediyl or C2-C8alkynediyl.

16. The PEG lipid according to any one of claims 1 to 15, wherein R2 is C7 to C 22 hydrocarbyl.

17. The PEG lipid of claim 16, wherein R2 is C7-C 22 alkyl, C7-C 22 alkenyl or C7-C 22 alkynyl.

18. [Amended according to Rule 26 25.11.2025] The PEG lipid according to any one of claims 1 to 17, wherein L1is a straight-chain C1-C8alkylene group, R2is a straight-chain C7-C 22 alkyl group.

19. The PEG lipid according to any one of claims 10-18, wherein -L1-R2 is C 16 ~C 22 alkyl.

20. The PEG lipid of claim 19, wherein -L1-R2 is a straight chain C 16 ~C 22 alkyl.

21. The PEG lipid of claim 19, wherein -L1-R2 is C 16 ~C 18 alkyl.

22. The PEG lipid of claim 21, wherein -L1-R2 is a straight chain C 16 ~C 18 alkyl.

23. [Amended according to Rule 26 25.11.2025] The PEG lipid according to any one of claims 1 to 6, 8 to 10, 12 to 22, wherein L2is C1-C3alkylene.

24. [Amended according to Rule 26 25.11.2025] The PEG lipid according to claim 23, wherein L2is C1-C3alkanediyl, C2-C3alkenediyl or C2-C3alkynediyl.

25. The PEG lipid according to claim 24, wherein L2 is a straight-chain C1-C3 alkylene, preferably C1 alkylene or C2 alkylene.

26. The PEG lipid according to any one of claims 1-6, 8-10, 12-26, wherein L3 is a C1-C2 alkylene.

27. The PEG lipid according to claim 26, wherein L3 is a C1-C2 alkylene, C2 alkenylene or C2 alkynylene.

28. The PEG lipid according to any one of claims 1-27, wherein n is any integer from 30 to 55.

29. The PEG lipid according to claim 28, wherein n is any integer from 35 to 50.

30. The PEG lipid according to claim 29, wherein n is any integer from 40 to 50.

31. The PEG lipid according to claim 30, wherein n is 45.

32. The PEG lipid of claim 1, wherein the PEG lipid is:

33. Use of a PEG lipid according to any one of claims 1-32 for the manufacture of a lipid nanoparticle.

34. A lipid nanoparticle comprising a PEG lipid according to any one of claims 1-32.

35. The lipid nanoparticle according to claim 34, wherein the lipid nanoparticle further comprises an active ingredient; wherein the active ingredient is preferably a pharmaceutically active ingredient, more preferably a nucleic acid. Preferably, the lipid nanoparticle further comprises one or more of a cationic lipid, a helper lipid, a structural lipid and a polymeric lipid.

36. A pharmaceutical composition comprising a lipid nanoparticle according to claim 34 or 35, and a pharmaceutically acceptable carrier.

37. Use of a lipid nanoparticle according to claim 34 or 35 or a pharmaceutical composition according to claim 36 for the manufacture of a medicament; Preferably, the medicament is for gene therapy, gene vaccination, protein replacement therapy, antisense therapy or therapy by interfering RNA; Preferably, the lipid particle is used as a carrier for the transfer or delivery of a pharmaceutically active ingredient; more preferably, the pharmaceutically active ingredient is a nucleic acid; Preferably, the medicament is for the treatment and / or prevention of a disease; Preferably, the medicament is for the treatment and / or prevention of a disease; Preferably, the medicament is for the treatment and / or prevention of one or more of the following: rare diseases, cancer, infectious diseases, autoimmune diseases, metabolic diseases, neurological diseases, cardiovascular diseases, transplant rejection, inflammatory reactions, genetic diseases and musculoskeletal diseases; Preferably, the medicament is a nucleic acid medicament, wherein the nucleic acid comprises at least one of the following: RNA, messenger RNA (mRNA), antisense oligonucleotide, DNA, plasmid, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA) and Cas9 mRNA.

38. [Amended according to Rule 26 25.11.2025] Use of the lipid nanoparticle of claim 34 or 35 for the manufacture of a carrier for the transfer or delivery of an active ingredient; Preferably, the active ingredient is a pharmaceutically active ingredient; Preferably, the pharmaceutically active ingredient is a nucleic acid; Preferably, the nucleic acid comprises at least one of the following: RNA, messenger RNA (mRNA), antisense oligonucleotide, DNA, plasmid, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA) and Cas9 mRNA.

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