Vaccine carrier composition, nano vaccine pharmaceutical composition, vaccine preparation and preparation method thereof

Abstract

The invention relates to a vaccine carrier composition, a nano vaccine pharmaceutical composition, a vaccine preparation and a preparation method thereof. The vaccine carrier composition is prepared from the following components in percentage by mass: 30 to 60 percent of lipid derivative DOPCO of targeted dendritic cells, 1 to 55 percent of auxiliary lipid, 0 to 15 percent of adjuvant component and 0 to 15 percent of other pharmaceutic adjuvants. The nano vaccine pharmaceutical composition comprises an antigen component and a vaccine carrier composition. The vaccine preparation comprises the nano vaccine pharmaceutical composition. The vaccine carrier composition, the nano vaccine pharmaceutical composition and the vaccine preparation have the characteristic of targeting dendritic cells, and the obtained nano vaccine has the capability of inducing generation of wide cross reaction antibodies and cellular immunity, can target lymph node dendritic cells and is efficiently taken by the dendritic cells; therefore, the antigen-specific immunoreaction is induced, and the method has a wide application prospect.

Description

Technical Field

[0001] This application relates to the field of biopharmaceutical technologies, and particularly relates to vaccine vector compositions, nano-vaccine pharmaceutical compositions, vaccine preparations and their preparation methods. Background Art

[0002] Given the high incidence and intractability of cancer, and the single effect of immunoprophylaxis against infectious diseases, there is an urgent need to develop new vaccines that can activate effective and persistent immune responses. Currently, the design and preparation of vaccines have evolved from initially only setting simple antigen components to using antigens as the main components, and also adding various immunoadjuvants and delivery materials. More and more studies have shown that even if antigens activate humoral immunity and produce neutralizing antibodies against foreign pathogens, the ability of single-component antigens to activate cellular immunity is still weak. Compared with the neutralizing antibodies produced by humoral immunity, the activation of cellular immunity plays a crucial role in immune defense and treatment. Therefore, the design of delivery materials and the addition of adjuvants are playing a vital role in the activation of cellular immunity.

[0003] Vaccines with a single antigen component, such as inactivated virus vaccines, are pathogens (such as viruses) that have been killed by physical or chemical methods. The genetic material (nucleic acid) of the virus used in inactivated vaccines has been destroyed, so they cannot replicate, but they can still stimulate the immune response of the human body. In such vaccines, the single antigen component results in low protection and a time-limited immune protection effect. Aluminum-containing adjuvants have been used for more than 90 years to enhance the immune response to vaccines, and the commonly used one is Al(OH)3 adjuvant. Aluminum salt adjuvants are called the "gold standard" against which all new adjuvants can be compared. However, vaccines containing aluminum salt adjuvants are prone to cause injection site reactions, and their ability to induce cellular immunity is weak. Factors such as efficacy and biodegradability also limit their application to a certain extent.

[0004] Currently, adding new adjuvants to vaccines has become a research and development hotspot, and new vaccine adjuvants are developing in diversified directions such as Toll-like receptor activators, composite adjuvants and nanoparticles. Common new adjuvants include adjuvant system AS03, CpG1018, etc. Currently, some new adjuvants have been widely used in the preparation of clinical vaccines, such as the influenza vaccine Pandemrix and the hepatitis B vaccine Heplisav-B. With the increasingly mature research and development of new adjuvants, they can also be prepared in the form of liposomes to improve their delivery effect. The new adjuvant Matrix-M has also been added to the COVID-19 vaccine NVX-CoV2373 developed by Nanovax. TM, which consists of two particles sized 40 nanometers, is formulated with purified saponin QS-21 from Gleditsia sinensis together with cholesterol and phospholipids, and has currently been put into clinical phase 1 / 2 trials with excellent data. Moreover, the carrier design of the novel vaccine carrier is also crucial for vaccine delivery. Patent document CN111658780B discloses a novel vaccine adjuvant in the form of nanoparticles, whose carrier material design is based on aluminum salts of anionic polymers and their derivatives, and can generate an effective immune response by promoting the uptake of the vaccine by immune cells. The antigens used can be various components, such as protein antigens, viral or bacterial lysates, and Toll-like receptor agonists, NOD-like receptor agonists, STING agonists, saponins, etc. can also be added to the adjuvant components. Based on the above vaccine research and development, although it can activate certain immune responses in the body, it lacks targeting and is difficult to achieve precise delivery, which limits the exertion of the maximum immune effect. Summary of the Invention

[0005] Based on this, the present application provides at least a vaccine carrier composition, a nano-vaccine pharmaceutical composition, a vaccine preparation and their preparation methods. The vaccine carrier composition, the nano-vaccine pharmaceutical composition containing the vaccine carrier composition and the vaccine preparation have the property of targeting dendritic cells. The obtained nano-vaccine has the ability to induce the production of broad cross-reactive antibodies and cellular immunity, can target lymph node dendritic cells and be efficiently taken up by dendritic cells, thereby inducing antigen-specific immune responses, and has broad application prospects.

[0006] In the first aspect of the present application, there is provided a vaccine carrier composition, which comprises the following components in mass percentages: 30% - 60% of a lipid derivative DOPCO targeting dendritic cells, 1% - 55% of auxiliary lipids, 0% - 15% of adjuvant components and 0% - 15% of other pharmaceutical excipients;

[0007] Among them, the structure of the lipid derivative DOPCO targeting dendritic cells is as follows:

[0008] Lipid structural unit - L 10 - Polyethylene glycol structural unit - L 20 - Chitosan oligosaccharide structural unit; where L 10 and L 20 are each independently a divalent linking group;

[0009] The lipid structural unit has the structure shown by formula U 1a as shown:

[0010] Among them, R 11 and R 12 are each independently a hydrocarbon group having 13 to 17 carbon atoms, and R 11 and R12 each independently contains 0, 1 or 2 carbon-carbon double bonds;

[0011] the number-average molecular weight of the polyethylene glycol structural unit is 1000 Da to 8800 Da;

[0012] the number-average molecular weight of the chitosan oligosaccharide structural unit is 340 Da to 3200 Da.

[0013] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO has the general structural formula shown in Formula Id:

[0014]

[0015] wherein k is an integer selected from 2 to 20; R 3 is a hydrogen atom or an acetyl group.

[0016] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO satisfies one or more of the following characteristics:

[0017] the lipid structural unit and the polyethylene glycol structural unit are connected by an amide bond;

[0018] the polyethylene glycol structural unit is connected to the amino site in the chitosan oligosaccharide structural unit through a carbamate group;

[0019] R 11 and R 12 are each independently a hydrocarbon group having 13 to 17 carbon atoms and having one carbon-carbon double bond;

[0020] the number-average molecular weight of the polyethylene glycol structural unit is 1500 Da to 3000 Da;

[0021] the number-average molecular weight of the chitosan oligosaccharide structural unit is 100 Da to 1700 Da;

[0022] the number-average molecular weight of the dendritic cell-targeting lipid derivative DOPCO is 1842 Da to 11818 Da;

[0023] the mass ratio of the lipid structural unit in the dendritic cell-targeting lipid derivative is 25% to 55%;

[0024] the mass ratio of the polyethylene glycol structural unit in the dendritic cell-targeting lipid derivative is 30% to 60%;

[0025] the mass ratio of the chitosan oligosaccharide structural unit in the dendritic cell-targeting lipid derivative is 25.33% to 55%.

[0026] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO has a general structural formula shown in Formula Ia:

[0027]

[0028] L 11 and L 21 are each independently an alkylene group having 1 to 4 carbon atoms;

[0029] m is an integer selected from 22 to 200;

[0030] COS is a chitosan oligosaccharide structural unit connected through its amino site.

[0031] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO has a general structural formula shown in Formula Ib:

[0032]

[0033] wherein, R 11 and R 12 are each independently a hydrocarbon group having 17 carbon atoms and one carbon-carbon double bond;

[0034] L 11 is methylene;

[0035] n is an integer selected from 23 to 200.

[0036] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO has a general structural formula shown in Formula Id:

[0037]

[0038] wherein, k is an integer selected from 1 to 20; R3 is a hydrogen atom or an acetyl group.

[0039] In some embodiments, the dendritic cell-targeting lipid derivative DOPCO satisfies one or more of the following characteristics:

[0040] n is an integer selected from 34 to 68;

[0041] k is an integer selected from 2 to 10.

[0042] In some embodiments, the vaccine carrier composition satisfies one or more of the following characteristics:

[0043] The vaccine carrier composition comprises the following components in mass percentages: 40% to 60% of a lipid derivative DOPCO targeting dendritic cells, 10% to 50% of an auxiliary lipid, 2.5% to 15% of an adjuvant component, and 4.5% to 15% of other pharmaceutical excipients;

[0044] The number-average molecular weight of the lipid derivative DOPCO targeting dendritic cells is 3008 Da to 6620 Da;

[0045] The auxiliary lipid is selected from one or more of the following lipid components: hydrogenated soy phosphatide, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, PEG-DSPE, and cholesterol; wherein, PEG-DSPE is a polyethylene glycolated derivative of distearoyl phosphatidylethanolamine, and the number-average molecular weight of the PEG part in the PEG-DSPE is 1000 Da to 2000 Da;

[0046] The adjuvant component is selected from one or more of the following components: Toll-like receptor agonists, Sting agonists, saponin adjuvants, aluminum adjuvants, Freund's adjuvants, poly(ethylene glycol)-poly(lactic-co-glycolic acid) copolymers, and squalene; the Toll-like receptor agonists include at least one of single-stranded mRNA, peptidoglycan, monophosphoryl lipid A, imiquimod, CpG oligonucleotides, bacterial flagellin, polyinosinic:polycytidylic acid, and lipoteichoic acid; the Sting agonists include at least one of CDNs and cGAMP; the saponin adjuvants include at least one of QS-21 and Quil-A; the aluminum adjuvants include at least one of aluminum phosphate, aluminum hydroxide, and potassium alum; the concentration of squalene is 0.5 wt% to 2.5 wt%;

[0047] The other pharmaceutical excipients are selected from one or more of the following pharmaceutical excipients: surfactants, emulsifiers, sustained-release agents, solubilizers, buffers, osmotic pressure regulators, antibacterial agents, protein stabilizers, vaccine protectants, and fillers.

[0048] In a second aspect of the present application, a nano-vaccine pharmaceutical composition is provided, which comprises an antigen component and the vaccine carrier composition described in the first aspect of the present application.

[0049] In some embodiments, the nano-vaccine pharmaceutical composition satisfies one or more of the following characteristics:

[0050] The mass ratio of the antigen component to the vaccine carrier is 0.1 to 10;

[0051] The antigen component is a recombinant protein, mRNA, inactivated virus, attenuated virus, or polypeptide;

[0052] The antigen component is at least one of tumor-associated antigens and infectious disease-related antigens; wherein, the sources of the tumor-associated antigens include at least one of the following: whole cell lysates of tumor cells, tumor cell membranes, tumor cell membranes and tumor cell exosomes; the infectious disease-related antigens include at least one of novel coronavirus antigen, influenza virus, hepatitis virus, infectious intestinal diseases, pulmonary tuberculosis, neonatal tetanus, mumps and chickenpox.

[0053] In the third aspect of the present application, a vaccine preparation is provided, which comprises the nano-vaccine pharmaceutical composition described in the second aspect of the present application;

[0054] The vaccine preparation is a solid preparation or a liquid preparation; wherein, the liquid preparation contains vaccine particles, and the average particle size of the vaccine particles is 30nm to 1000nm;

[0055] In the liquid preparation, at least a part of the vaccine carrier composition forms a vesicle structure, and at least a part of the antigen component is encapsulated in the internal cavity of the vesicle structure or hybridized on the surface of the vesicle structure;

[0056] When the vaccine preparation is a solid preparation, the solid preparation can be reconstituted to form the liquid preparation.

[0057] In some embodiments, the vaccine preparation satisfies one or more of the following characteristics:

[0058] The liquid preparation contains vaccine particles, and the average particle size of the vaccine particles is 100nm to 1000nm, and can be 100nm to 500nm;

[0059] The liquid preparation contains a dispersion liquid; the dispersion liquid includes water, and may or may not include a buffer; wherein, the buffer includes one or more of the salt components of phosphate buffer solution, tris(hydroxymethyl)methylaminopropanesulfonic acid, sodium citrate, citric acid, sodium dihydrogen phosphate, potassium dihydrogen phosphate and 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid.

[0060] In the fourth aspect of the present application, a preparation method of a vaccine preparation is provided, which can be used to prepare the vaccine preparation described in the third aspect of the present application, and comprises the following steps:

[0061] The liquid preparation is prepared by any one of the following methods:

[0062] Method 1: The vaccine preparation is prepared by the thin film hydration method using the nano-vaccine pharmaceutical composition and the antigen component; wherein, the antigen component includes non-nucleic acid antigens;

[0063] Method 2: The vaccine preparation is prepared by microfluidic assembly of the nano-vaccine pharmaceutical composition and the antigen component; wherein, the antigen component includes nucleic acid antigens; and

[0064] Method 3: The vaccine preparation is prepared by pressure homogenization and emulsification of the nano-vaccine pharmaceutical composition and the antigen component;

[0065] The preparation method optionally includes the step of drying the liquid preparation into a solid preparation.

[0066] In the fifth aspect of the present application, a lipid derivative targeting dendritic cells is provided, which is the lipid derivative DOPCO targeting dendritic cells described in the first aspect of the present application.

[0067] In the sixth aspect of the present application, there is provided the use of the vaccine carrier composition described in the first aspect of the present application, or the nano-vaccine pharmaceutical composition described in the second aspect of the present application, or the lipid derivative targeting dendritic cells described in the fifth aspect of the present application in the preparation of a nano-vaccine preparation targeting dendritic cells.

[0068] In the seventh aspect of the present application, there is provided the use of the vaccine carrier composition described in the first aspect of the present application, or the nano-vaccine pharmaceutical composition described in the second aspect of the present application, or the vaccine preparation described in the third aspect of the present application, or the vaccine preparation prepared by the preparation method described in the fourth aspect of the present application in the preparation of a drug for preventing and / or treating at least one of tumors and infectious diseases.

[0069] In the vaccine carrier composition provided in the first aspect of the present application, a lipid derivative DOPCO targeting dendritic cells (which may also be abbreviated as DOPCO, DOPCO lipid derivative, etc. in this article) is introduced. DOPCO is a cationic lipid, which includes a lipid structural unit (which may also be called a lipid tail chain), a polyethylene glycol structural unit, and a chitosan oligosaccharide structural unit (also denoted as a chitosan oligosaccharide head group) connected in sequence; the chitosan oligosaccharide structural unit therein can interact with the mannose receptor on the surface of dendritic cells (which may be denoted as DC or DC cells), enabling DOPCO to efficiently target DC cells in the body and exhibit a targeting and retention effect at the draining lymph nodes; the lipid structural unit includes a fatty acid chain and an ionizable amino group, having ionizable properties; the lipid structural unit and the chitosan oligosaccharide structural unit are covalently connected through the polyethylene glycol structural unit as an intermediate connecting unit, enhancing the stability of DOPCO and enabling it to improve the immune activation ability. DOPCO has the characteristics of being ionizable, having a stable structure, being able to target the mannose receptor on dendritic cells, and having immune activation.

[0070] The vaccine preparation containing DOPCO can be prepared using the aforementioned vaccine carrier composition or the nano-vaccine pharmaceutical composition containing the vaccine carrier composition. It can form a nano-vaccine and endow the nano-vaccine with the property of targeting dendritic cells, effectively improving the antigen delivery efficiency of the vaccine, enhancing antigen immunogenicity, and increasing the uptake efficiency of dendritic cells, thereby optimizing the vaccine treatment plan. This nano-vaccine targeting dendritic cells can be combined with a variety of traditional or novel antigen components (such as inactivated virus antigens, single protein / polypeptide antigens of viruses or tumors, whole cell lysate antigens, cancer cell autologous cell membrane antigens, etc.), and can also introduce a variety of novel or less toxic adjuvants to achieve the co-delivery of antigens and adjuvants, significantly stimulating DC maturation, generating a stronger T cell immune response, and effectively increasing the level of neutralizing antibodies in vivo and inducing long-term immune memory.

[0071] The vaccine carrier composition provided in this application and the nano-vaccine pharmaceutical composition containing the vaccine carrier composition and the vaccine preparation have the property of targeting dendritic cells. The obtained nano-vaccine is a targeted nano-vaccine that can target dendritic cells, has the ability to induce the production of a wide range of cross-reactive antibodies and cellular immunity, can target lymph node dendritic cells and be efficiently taken up by dendritic cells, thereby inducing an antigen-specific immune response, and has broad application prospects.

[0072] The targeted nano-vaccine provided in this application can be used for at least one of the prevention and treatment of diseases such as tumors (such as cancer), infectious diseases, etc., for example, it can be used for tumor immunity, prevention and / or treatment of infections, etc. In addition, the design of the nano-vaccine targeting dendritic cells provided in this application is of great significance for the development of universal tumor or infectious disease vaccines. Brief Description of the Drawings

[0073] In order to more clearly illustrate the technical solutions in the embodiments and implementations of this application and to more fully understand this application and its beneficial effects, the drawings required for use in the description of the embodiments or implementations will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of this application. For those skilled in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0074] Figure 1 It is the particle size and potential characterization results of different formulations in some embodiments of this application.

[0075] Figure 2 It is the characterization result of different formulations targeting DC cells in vivo and being taken up in some embodiments of this application.

[0076] Figure 3Visualization characterization results of different formulations being taken up by in vivo targeted DC cells in some embodiments of the present application.

[0077] Figure 4 Visualization characterization results of lysosomal escape of DC cells in vivo with different formulations in some embodiments of the present application.

[0078] Figure 5 Characterization results of the percentages of CD11c+ / CD86+ DC cells and CD11c+ / CD80+ DC cells in vivo with different formulations in some embodiments of the present application.

[0079] Figure 6 Graphs of interleukin-12 secretion levels and tumor necrosis factor secretion levels of different formulations in some embodiments of the present application.

[0080] Figure 7 Graphs of CD8+ T cell activation levels and CD4+ T cell activation levels of different formulations in some embodiments of the present application.

[0081] Figure 8 Tumor growth curves under different formulations in some embodiments of the present application.

[0082] Figure 9 Efficiency of neutralizing pseudovirus with different formulations in some embodiments of the present application. Specific Embodiments

[0083] The present application will be further described in detail below in conjunction with the accompanying drawings, embodiments and examples. It should be understood that these embodiments and examples are only used to illustrate the present application and not to limit the scope of the present application. The purpose of providing these embodiments and examples is to make the understanding of the disclosed content of the present application more thorough and comprehensive. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various changes or modifications without departing from the connotation of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. For example, features described or illustrated as part of one embodiment can be combined in a suitable manner with another embodiment to produce a new embodiment. In addition, in the following description, a large number of details are given to provide a more thorough understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0084] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which this application belongs. The terms used in the description of the present application in this specification are only for the purpose of describing the embodiments and examples and are not intended to limit the present application.

[0085] Terms

[0086] Unless otherwise specified or there are contradictions, the terms or phrases used in this application have the following meanings:

[0087] In this application, the terms "multiple", "diverse", "multiple times", "multiple items", "several", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0088] In this application, unless otherwise stated, "one or more" means any one of the listed items or any combination of the listed items. Similarly, in other cases where "one or more" is expressed in other ways, such as "one or multiple", unless otherwise stated, the same understanding applies.

[0089] In this application, the "suitable combination method", "suitable method", "any suitable method", etc., the "suitable" is based on being able to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0090] In this application, "preferred", "better", "more preferred", "should preferably be", "relatively better", "more preferred" are only used to describe embodiments or examples with better effects. It should be understood that they do not constitute a limitation on the protection scope of this application. If there are multiple "preferred" in a technical solution, unless otherwise specified, and there are no contradictions or mutual restrictions, each "preferred" is independent.

[0091] In this application, "further", "even further", "especially", "for example", "such as", "example", "for illustration" are used for descriptive purposes, indicating that there is a connection in the content covered between the different technical solutions before and after, but it should not be understood as a limitation on the previous technical solution, nor can it be understood as a limitation on the protection scope of this application. In this application, unless otherwise stated, A (such as B) means that B is a non-limiting example of A, and it can be understood that A is not limited to B.

[0092] In this application, "optionally", "optional", "optional" mean that it can be either present or absent, that is, it refers to any one of the two alternative options of "present" or "absent". If there are multiple "optional" in a technical solution, unless otherwise specified, and there are no contradictions or mutual restrictions, each "optional" is independent. Unless otherwise stated, in this application, descriptions such as "optionally include", "optionally contain", etc., taking "optionally include" as an example, mean "can include or not include". "Optional component X" means that component X is either present or absent.

[0093] As used in this application, the terms "comprising", "including" and "containing" are synonyms, which are inclusive or open-ended and do not exclude additional, unrecited members or features. The members or features include, for example, materials or components, structures, elements, instruments, etc.; non-limiting examples of members or features also include actions, conditions under which actions occur, timing, states, etc.

[0094] In this application, among the technical features or technical solutions described in open language, those that include the recited content as a closed technical feature or technical solution are also included, as well as those that include the recited content as an open technical feature or technical solution.

[0095] In this application, the terms "room temperature" and "normal temperature" generally refer to 4°C to 35°C, such as 20°C ± 5°C. In some embodiments of this application, "room temperature" or "normal temperature" refers to 20°C to 30°C.

[0096] In this application, for units related to data ranges, if the unit is only attached after the right endpoint, it means that the units of the left endpoint and the right endpoint are the same. For example, 3 to 5 h means that the units of the left endpoint "3" and the right endpoint "5" are both h (hours), and it has the same meaning as 3 h to 5 h. In addition, the above understanding method also applies to similar descriptions of other parameters such as temperature and size.

[0097] All documents mentioned in this application are incorporated herein by reference as if each document was individually incorporated by reference. Unless it conflicts with the inventive purpose and / or technical solution of this application, the cited documents involved in this application are cited for their entire content and entire purpose. When this application involves cited documents, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also incorporated by reference. When this application involves cited documents, examples and preferred methods of the relevant technical features cited can also be incorporated as references into this application, but only to the extent that this application can be implemented. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or be amended adaptively according to the description in this application.

[0098] In this application, for a method process involving multiple steps, unless there are clear different descriptions in this article, the execution of these steps has no strict order limit, and they can be executed in other orders than those described. Moreover, any one step may include multiple sub-steps or multiple stages. These sub-steps or stages do not necessarily need to be executed at the same time, but can be executed at different times, and their execution order does not necessarily need to be sequential, but can be executed alternately or simultaneously with other steps or a part of the sub-steps or stages of other steps.

[0099] In the present application, the exemplary descriptions such as "in some embodiments" or "in one embodiment" may cover, but are not limited to, the following meanings: These solutions can be combined with other solutions in a suitable manner to form new technical solutions.

[0100] In the present application, in "the first aspect", "the second aspect", "the third aspect", "the fourth aspect", etc., the terms "first", "second", "third", "fourth", etc. are only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumerative description and should be understood not to constitute a closed limitation on quantity.

[0101] In the present application, regarding the numerical interval (i.e., the numerical range), unless otherwise specified, the distribution of the selectable numerical values within this numerical interval is considered continuous and includes the two numerical endpoints of this numerical interval (i.e., the minimum value and the maximum value), as well as each numerical value between these two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within this numerical interval, it includes the two endpoint integers of this numerical range, as well as each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all sub-ranges subsumed therein. The "numerical value" in this numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. The "numerical interval" allows for a broad inclusion of numerical interval types such as percentage intervals, ratio intervals, ratio value intervals, etc.

[0102] In the present application, unless otherwise stated, the "hydrocarbyl group" is a group composed of hydrogen atoms and carbon atoms and can be a monovalent group, a divalent group or a polyvalent group. Unless otherwise specified, the hydrocarbyl group involved refers to a monovalent hydrocarbyl group.

[0103] The present application provides at least one vaccine carrier composition, a nano-vaccine pharmaceutical composition, a vaccine preparation and a preparation method thereof. The nano-vaccine pharmaceutical composition includes an antigen component and a vaccine carrier composition. The vaccine preparation includes the nano-vaccine pharmaceutical composition. The vaccine carrier composition, the nano-vaccine pharmaceutical composition and the vaccine preparation have the property of targeting dendritic cells. The obtained nano-vaccine has the ability to induce the production of a wide range of cross-reactive antibodies and cellular immunity, can target lymph node dendritic cells and be efficiently taken up by dendritic cells, thereby inducing an antigen-specific immune response and having broad application prospects.

[0104] In a first aspect of the present application, there is provided a vaccine carrier composition, which comprises a lipid derivative DOPCO targeting dendritic cells and a co-lipid. Among them, the mass percentage of the lipid derivative DOPCO targeting dendritic cells in the vaccine carrier composition can be 30% to 60%, and the mass percentage of the co-lipid in the vaccine carrier composition can be 1% to 55%.

[0105] In some embodiments, the vaccine carrier composition comprises components in the following mass percentages: 30% to 60% of the lipid derivative DOPCO targeting dendritic cells, 1% to 55% of the co-lipid, 0% to 15% of an adjuvant component, and 0% to 15% of other pharmaceutical excipients.

[0106] In some embodiments, the vaccine carrier composition comprises the following components in the following mass percentages: 40% to 60% of the lipid derivative DOPCO targeting dendritic cells, 10% to 50% of the co-lipid, 2.5% to 15% of the adjuvant component, and 4.5% to 15% of other pharmaceutical excipients.

[0107] In the present application, DOPCO, the co-lipid, and the optional adjuvant component are all used as pharmaceutical excipients.

[0108] In the present application, unless otherwise specified, "vaccine carrier composition" refers to a composition for providing a vaccine carrier.

[0109] In the present application, unless otherwise specified, "other pharmaceutical excipients" refers to pharmaceutical excipients other than DOPCO, the co-lipid, and the optional adjuvant component (if any).

[0110] It can be understood that any pharmaceutical excipient in the present application is a pharmaceutically acceptable pharmaceutical excipient. Among them, the selection range of the co-lipid, the optional adjuvant component, and other pharmaceutical excipients can be appropriately selected within the range permitted by administrative regulations. Those skilled in the art can appropriately select relevant raw materials according to the administrative regulations during implementation, and can also exclude those prohibited components according to the prohibitive regulations.

[0111] In some embodiments, the structure of the lipid derivative DOPCO targeting dendritic cells is as follows:

[0112] Lipid structural unit - L 10 - Polyethylene glycol structural unit - L 20 - Chitosan oligosaccharide structural unit; wherein, L 10 and L 20 are each independently a divalent linking group;

[0113] The lipid structural unit has the structure shown in formula U 1a as shown:

[0114] wherein, R 11 and R 12 are each independently a hydrocarbon group having 13 to 17 carbon atoms, and R 11 and R 12 each independently contain 0, 1 or 2 carbon-carbon double bonds.

[0115] In some embodiments, the number-average molecular weight of the polyethylene glycol structural unit does not exceed 10 kDa, and for example, it can be 1000 Da to 8800 Da.

[0116] In some embodiments, the number-average molecular weight of the chitosan oligosaccharide structural unit can be 340 Da to 3200 Da.

[0117] In the vaccine carrier composition provided in the first aspect of the present application, a lipid derivative DOPCO (which can also be abbreviated as DOPCO, DOPCO lipid derivative, etc. in this article) targeting dendritic cells is introduced. DOPCO is a cationic lipid, which includes a lipid structural unit (which can also be called a lipid tail chain), a polyethylene glycol structural unit, and a chitosan oligosaccharide structural unit (also denoted as a chitosan oligosaccharide head group) connected in sequence; the chitosan oligosaccharide structural unit therein can interact with the mannose receptor on the surface of dendritic cells (dentritic cell, which can be denoted as DC or DC cell), enabling DOPCO to efficiently target DC cells in the body and showing a targeting and retention effect at the draining lymph nodes; the lipid structural unit therein includes a fatty acid chain and an ionizable amino group, having an ionizable property; a polyethylene glycol structural unit is used as an intermediate linking unit between the lipid structural unit and the chitosan oligosaccharide structural unit to achieve covalent bonding, enhancing the stability of DOPCO and improving the immune activation ability. DOPCO has the characteristics of being ionizable, having a stable structure, being able to target the mannose receptor on dendritic cells, and having immune activation.

[0118] In the present application, the chitooligosaccharide (COS) structural unit is a monovalent residue formed by removing one amino hydrogen atom from chitooligosaccharide, which serves as the head group of DOPCO. The chemical name of chitooligosaccharide is β-1,4-2-amino-2-deoxy-D-glucosamine. It is a bioactive cationic polysaccharide obtained by deacetylation of chitin. It is rich in sources and simple to prepare. It is the only basic polysaccharide existing in nature and has physicochemical properties such as good biocompatibility and natural degradability. The chitooligosaccharide structural unit can interact with the mannose receptor on the surface of dendritic cells (DC cells), enabling DOPCO to exhibit the characteristic of efficiently targeting DC cells in vivo and having a targeting and retention effect at the draining lymph nodes. In addition, it endows DOPCO with medical effects such as anti-inflammatory, antibacterial and immune enhancement, and can play an important role in regulating innate immunity and specific immunity. It has an immune activation effect and can also participate in various signal pathways related to inflammatory responses (such as the NF-κB signal pathway).

[0119] If chitosan with a higher molecular weight is used (such as a molecular weight greater than 5000 Da), it will lead to poor water solubility, be difficult to be absorbed by the body, and not have the mannose targeting property.

[0120] In some embodiments, the number-average molecular weight of the chitooligosaccharide structural unit is 340 Da to 3200 Da, and can be optionally 100 Da to 1700 Da. The number of pyranose rings in the chitooligosaccharide structural unit can be 2 to 21, further can be 2 to 20, still further can be 2 to 10, and can also be any one of the following values or an interval composed of any two of the following values: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.

[0121] In some embodiments, the mass percentage of the chitooligosaccharide structural unit in the lipid derivative targeting dendritic cells is 25.33% to 55%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 25.33%, 26%, 28%, 30%, 35%, 40%, 45%, 50%, 55%, etc.

[0122] In some embodiments, the chitooligosaccharide structural unit has the formula U 3a as shown in the structure:

[0123]

[0124] wherein, k is an integer selected from 1 to 20; R3 is a hydrogen atom or an acetyl group (-C(=O)CH3).

[0125] In some embodiments, k is an integer selected from 1 to 20, can be an integer selected from 2 to 20, further can be an integer selected from 2 to 10, and can also be any one of the following integers or an interval composed of any two of the following integers: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.

[0126] In some embodiments, R3 is an acetyl group. In some other embodiments, R3 is a hydrogen atom.

[0127] Non - restrictively, in the vaccine carrier composition, each occurrence of R3 can independently be a hydrogen atom or an acetyl group. In some embodiments, in the vaccine carrier composition, R3 is a hydrogen atom. In some other embodiments, in the vaccine carrier composition, R3 is an acetyl group. In some other embodiments, in the vaccine carrier composition, R3 is a combination of a hydrogen atom and an acetyl group.

[0128] In the present application, the lipid structural unit includes a fatty acid chain and an ionizable amino group (-NH-), has ionizable properties, and is an ionizable lipid structure. The lipid structural unit enables DOPCO to maintain a neutral or low cationic surface charge density under physiological pH conditions, can minimize toxicity, minimize the degradation of plasma proteins, and in addition, can efficiently load nucleic acids under low pH conditions.

[0129] The lipid structural unit can have the structure of formula U 1a as shown:

[0130] wherein, R 11 and R 12 can each independently be a hydrocarbon group having 13 to 17 carbon atoms, and R 11 and R 12 can each independently contain 0, 1 or 2 carbon - carbon double bonds.

[0131] R 11 and R 12 can have the same or different structures. In some embodiments, R 11 and R 12 have the same structure.

[0132] In some embodiments, the molecular weight range of the lipid structural unit can be 502 Da to 618 Da.

[0133] R 11 and R 12 can each independently be a hydrocarbon group having 13 to 17 carbon atoms, so that the lipid structural unit carries a fatty acid chain having 14 to 18 carbon atoms. R 11 and R12 The number of carbon atoms in 11 and 12 can each independently be 13, 14, 15, 16 or 17, and can also be selected from the intervals formed by any two of the above integers. R 11 and 12 can each independently be a saturated hydrocarbon group or an unsaturated hydrocarbon group, and can contain 0, 1 or 2 carbon-carbon double bonds. In some embodiments,

[0134] In some embodiments, 11 and 12 are each independently a hydrocarbon group having 13 to 17 carbon atoms and having one carbon-carbon double bond.

[0135] In some embodiments, the lipid structural unit has the structure shown by formula U 1b as follows:

[0136] wherein, a1, a2, a3 and a4 are each independently an integer selected from 0 to 10.

[0137] In some embodiments, a1 and a3 are each independently an integer selected from 0 to 7. In some examples, a1 and a3 can both be 7. In other examples, a1 and a3 are both 4.

[0138] In some embodiments, a1 and a3 are each independently an integer selected from 0 to 5. In some examples, a1 and a3 can both be 5.

[0139] In some embodiments, the lipid structural unit has the structure shown by formula U 1c as follows:

[0140] At this time, the lipid structural unit is a monovalent residue formed by deaminating hydrogen of DOA (dioleostearin-3-amino-1,2-propanediol).

[0141] A covalent bond connection is achieved between the lipid structural unit and the chitosan oligosaccharide structural unit with a polyethylene glycol (PEG) structural unit as an intermediate linking unit, so that the stability of DOPCO is enhanced, the immune activation ability can be improved, and the immune stimulation effect can be better exerted.

[0142] The number-average molecular weight of the polyethylene glycol structural unit can be 1000 Da to 8800 Da, further can be 1000 Da to 8000 Da, further can be 1500 Da to 3000 Da, and can also be any one of the following molecular weights or an interval composed of any two of the following molecular weights: 1000 Da, 1100 Da, 1200 Da, 1300 Da, 1400 Da, 1500 Da, 1600 Da, 1800 Da, 2000 Da, 2200 Da, 2400 Da, 2500 Da, 2600 Da, 2800 Da, 3000 Da, 3400 Da, 3450 Da, 3500 Da, 3600 Da, 4000 Da, 4500 Da, 5000 Da, 6000 Da, 7000 Da, 8000 Da, 8800 Da, etc.

[0143] In some embodiments, the mass percentage of the lipid structural unit in the lipid derivative targeting dendritic cells is 25% to 55%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 25%, 26%, 28%, 30%, 35%, 40%, 45%, 50%, 55%, etc.

[0144] The polyethylene glycol structural unit includes a polyether chain segment formed by sequentially connecting ethylene oxide units (EO units, -CH2CH2O-), and this polyether chain segment can have the following structure The two * are respectively covalent bond connection sites; m can be an integer selected from 22 to 200, further can be an integer selected from 23 to 200, still further can be an integer selected from 34 to 73, still further can be an integer selected from 34 to 68, and can also be any one of the following integers or an interval composed of any two of the following integers: 22, 23, 24, 25, 26, 28, 30, 32, 33, 34, 35, 40, 44, 45, 50, 60, 68, 70, 80, 90, 91, 100, 110, 113, 114, 120, 130, 136, 137, 140, 150, 159, 160, 170, 180, 181, 182, 190, 199, etc.

[0145] In some embodiments, the mass percentage of the polyethylene glycol structural unit in the lipid derivative targeting dendritic cells is 30% to 60%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 30%, 35%, 40%, 45%, 50%, 55%, 60%, etc.

[0146] In some embodiments, the lipid structural unit and the polyethylene glycol structural unit are connected by an amide bond, and the stability is significantly improved compared with the ester bond connection (-C-O-C(=O)-C), achieving excellent immune activation ability.

[0147] In some embodiments, the polyethylene glycol structural unit is linked to the amino site in the chitosan oligosaccharide structural unit through a carbamate group. This linkage can enhance the stability of DOPCO and is beneficial to improving the immune activation ability.

[0148] In some embodiments, the lipid derivative DOPCO targeting dendritic cells has a general structural formula shown in Formula Ia:

[0149]

[0150] L 11 and L 21 are each independently an alkylene group having 1 to 4 carbon atoms; the value of m can be referred to the definition in the context. For example, m can be an integer selected from 22 to 200; COS is a chitosan oligosaccharide structural unit linked through its amino site.

[0151] L 11 and L 21 can each independently be methylene, ethylene, propylene or butylene, and further can each independently be methylene, 1,2-ethylene, 1,3-propylene or 1,4-butylene. In some embodiments, L 11 is methylene or ethylene. In some embodiments, L 11 is methylene. In other embodiments, L 11 is ethylene. In some embodiments, L 21 is ethylene. At this time, the number of EO units in the polyethylene glycol structural unit can be regarded as m + 1.

[0152] In Formula Ia, COS is a chitosan oligosaccharide structural unit linked through its amino site. At this time, a carbamate group linkage is formed between the chitosan oligosaccharide structural unit and the polyethylene glycol structural unit.

[0153] In some embodiments, COS can have the structure shown in Formula U 3a . In some of these embodiments, R3 in Formula U 3a is an acetyl group. In some other of these embodiments, R3 in Formula U 3a is a hydrogen atom. In other embodiments, R3 in the vaccine carrier composition is a combination of a hydrogen atom and an acetyl group.

[0154] In some embodiments, the lipid derivative DOPCO targeting dendritic cells has a general structural formula shown in Formula Ib:

[0155]

[0156] In Formula Ib, R 11 and R12 Each can independently refer to the definition in the context. Without limitation, R 11 and R 12 are each independently a hydrocarbyl group having 17 carbon atoms and having one carbon-carbon double bond.

[0157] In formula Ib, L 11 can refer to the definition in the context. Without limitation, L 11 can be methylene or ethylene. In some embodiments, L 11 is methylene. In other embodiments, L 11 is ethylene.

[0158] In formula Ib, n is an integer selected from 23 to 200, further can be an integer selected from 34 to 73, still further can be an integer selected from 34 to 68, and can also be any one of the following integers or an interval composed of any two of the following integers: 23, 24, 25, 26, 28, 30, 32, 33, 34, 35, 40, 44, 45, 50, 60, 68, 70, 80, 90, 91, 100, 110, 113, 114, 120, 130, 136, 137, 140, 150, 159, 160, 170, 180, 181, 182, 190, 199, etc.

[0159] In some embodiments, in formula Ib, R 11 and R 12 are each independently a hydrocarbyl group having 17 carbon atoms and having one carbon-carbon double bond; L 11 is methylene; n is an integer selected from 23 to 200.

[0160] When L 21 is 1,2-ethylene and n = m + 1, formula Ia and formula Ib have the same structure.

[0161] In some embodiments, the lipid derivative DOPCO targeting dendritic cells has the general formula structure shown in formula Ic:

[0162] The definition of n can refer to the definition in the context, as defined in formula Ib.

[0163] In some embodiments, the lipid derivative DOPCO targeting dendritic cells has the general formula structure shown in formula Id:

[0164]

[0165] Among them, the definitions of n, k, and R3 can be referred to the context. In some embodiments, n is an integer selected from 23 to 200 (optionally 34 to 68), k is an integer selected from 1 to 20 (optionally 2 to 20, further optionally 2 to 10); R3 is a hydrogen atom or an acetyl group. The definition of R3 can also be referred to the context.

[0166] In some embodiments, the number-average molecular weight of the lipid derivative DOPCO targeting dendritic cells is 1842 Da to 11818 Da, further optionally 2000 Da to 10000 Da, still further optionally 3008 Da to 6620 Da, and can also be any one of the following molecular weights or an interval composed of any two of the following molecular weights: 1842 Da, 2000 Da, 2200 Da, 2400 Da, 2500 Da, 2600 Da, 2800 Da, 3000 Da, 3400 Da, 3450 Da, 3500 Da, 3600 Da, 4000 Da, 4500 Da, 5000 Da, 6000 Da, 7000 Da, 8000 Da, 8800 Da, 10000 Da, 11818 Da, etc.

[0167] The helper lipid can be used to maintain the structural stability of the prepared nano-vaccine. Without limitation, the helper lipid can be selected from one or more of the following lipid components: hydrogenated soy phosphatidylcholine (HSPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), PEG-DSPE, and cholesterol; among them, PEG-DSPE is a polyethylene glycolated derivative of distearoyl phosphatidylethanolamine. Further, the number-average molecular weight of the PEG part in PEG-DSPE can be 1000 Da to 2000 Da. By introducing cholesterol, the fluidity can be adjusted.

[0168] As used herein, the "adjuvant component" refers to a substance used to enhance the potency of a vaccine. The immune-stimulating effect can be enhanced by introducing an adjuvant component. Without limitation, the adjuvant component can be selected from one or more of the following components: Toll-like receptor agonists, Sting agonists, saponin adjuvants, aluminum adjuvants, Freund's adjuvants, poly(ethylene glycol)-poly(lactic-co-glycolic acid) (PEG-PLGA), and squalene. Among them, the Toll-like receptor agonists can include, but are not limited to, at least one of single-stranded mRNA, peptidoglycan, MPLA (monophosphate lipid A), imiquimod, CpG oligonucleotides (CpG-ODN), bacterial flagellin, polyinosinic:polycytidylic acid (Poly I:C), and lipoarabinomannan. The Sting agonists can include, but are not limited to, at least one of CDNs and cGAMP. The saponin adjuvants can include, but are not limited to, at least one of QS-21 and Quil-A. The aluminum adjuvants can include, but are not limited to, at least one of aluminum phosphate, aluminum hydroxide, and potassium alum. The concentration of squalene can be, but is not limited to, 0.5 wt% to 2.5 wt% (such as 0.5 wt%, 1.0 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, etc.), which can improve immunogenicity.

[0169] Without limitation, other pharmaceutical excipients can be selected from one or more of the following pharmaceutical excipients: surfactants, emulsifiers, sustained-release agents, solubilizers, buffers, osmotic pressure regulators, antibacterial agents, protein stabilizers, vaccine protectants, and fillers. For example, other pharmaceutical excipients can include sodium chloride.

[0170] Without limitation, polysorbate 80, Span, etc. can be selected as surfactants; poloxamer can be selected as an emulsifier or a sustained-release agent; cyclodextrin can be selected as a solubilizer or a sustained-release agent; sodium chloride, magnesium chloride, etc. can be selected as buffers and pH regulators; sucrose, etc. can be selected as osmotic pressure regulators; glycerol, etc. can be selected as antibacterial agents; maltose can be selected as a protein stabilizer; mannitol can be selected as a vaccine protectant and a filler.

[0171] In some embodiments, the mass percentage of the lipid derivative DOPCO targeting dendritic cells in the vaccine carrier composition can be 30% to 60%, further can be 40% to 60%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 30%, 35%, 40%, 45%, 50%, 55%, 60%, etc.

[0172] In some embodiments, the mass percentage of the auxiliary lipid in the vaccine carrier composition can be 1% to 55%, further can be 10% to 50%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, etc.

[0173] In some embodiments, the mass percentage of the adjuvant component in the vaccine carrier composition can be 0% to 15%, further can be 2.5% to 15%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 0%, 1%, 1.5%, 2%, 2.5%, 3%, 4%, 5%, 6%, 8%, 10%, 12%, 15%, etc.

[0174] In some embodiments, the mass percentage of other pharmaceutical excipients in the vaccine carrier composition can be 0% to 15%, further can be 4.5% to 15%, and can also be any one of the following percentages or an interval composed of any two of the following percentages: 0%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 8%, 10%, 12%, 15%, etc.

[0175] In some embodiments, the vaccine carrier composition comprises components in the following mass percentages: 30% to 60% (optionally 40% to 60%) of the lipid derivative DOPCO targeting dendritic cells, 1% to 55% (optionally 10% to 50%) of the auxiliary lipid, 0% to 15% (optionally 2.5% to 15%) of the adjuvant component, and 0% to 15% (optionally 4.5% to 15%) of other pharmaceutical excipients. The mass percentages of each component can also refer to the definitions in the context.

[0176] In the second aspect of the present application, a nano-vaccine pharmaceutical composition is provided, which comprises an antigen component and the vaccine carrier composition described in the first aspect of the present application.

[0177] After the nano-vaccine containing DOPCO and the antigen component is ingested into the body, when it reaches the intracellular lysosome site, the low pH condition makes DOPCO exhibit cationic lipid characteristics. After endocytosis, the cationic lipid interacts with the anionic phospholipids naturally present in the endosome membrane to form an ion pair with a non-bilayer structure and disrupt the membrane, so that the encapsulated antigen component undergoes lysosomal escape in the cell and exerts a better delivery effect.

[0178] The aforementioned vaccine carrier composition or the nano-vaccine pharmaceutical composition containing the vaccine carrier composition can be used to prepare a vaccine preparation containing DOPCO, which can form a nano-vaccine and endow the nano-vaccine with the property of targeting dendritic cells, effectively improving the antigen delivery efficiency of the vaccine, enhancing antigen immunogenicity, and increasing the uptake efficiency of dendritic cells, thereby optimizing the vaccine treatment plan. The nano-vaccine targeting dendritic cells can be combined with a variety of traditional or novel antigen components (such as inactivated virus antigens, single protein / polypeptide antigens of viruses or tumors, whole cell lysate antigens, autologous cancer cell membrane antigens, etc.), and can also introduce a variety of novel or less toxic and side-effect adjuvants to achieve the co-delivery of antigens and adjuvants, significantly stimulating DC maturation, generating a stronger T cell immune response, effectively increasing the level of neutralizing antibodies in vivo and inducing long-term immune memory.

[0179] In some embodiments, the mass ratio of the antigen component to the vaccine carrier is 0.1 to 10, and can also be any one of the following ratios or an interval composed of any two of the following ratios: 0.1, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 2 / 3, 0.75, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, 20 / 3, 7, 7.5, 8, 9, 9.9, 10, etc.

[0180] Non-limitingly, the antigen component can be a recombinant protein, mRNA, inactivated virus, attenuated virus or polypeptide.

[0181] The antigen component can be, but is not limited to, at least one of tumor-associated antigens and infectious disease-related antigens.

[0182] In some embodiments, the source of the tumor-associated antigen can include, but is not limited to, at least one of the following: tumor cell whole cell lysate, tumor cell membrane, tumor cell membrane and tumor cell exosomes. The tumor can include, but is not limited to, at least one of triple-negative breast cancer, pancreatic cancer and colorectal cancer.

[0183] In some embodiments, the infectious diseases include at least one of novel coronavirus infection, influenza and hepatitis, infectious intestinal diseases, pulmonary tuberculosis, neonatal tetanus, mumps, and chickenpox. The infectious disease-related antigens can include, but are not limited to, at least one of new coronavirus antigens, influenza viruses, hepatitis viruses, infectious intestinal diseases, pulmonary tuberculosis, neonatal tetanus, mumps, and chickenpox. Non-limiting examples of new coronavirus antigens can be, for example, RBD domain recombinant proteins, S proteins, mRNA, polypeptides, etc.

[0184] In the third aspect of the present application, a vaccine preparation is provided, which includes the nano-vaccine pharmaceutical composition described in the second aspect of the present application. The vaccine preparation can be a solid preparation or a liquid preparation.

[0185] In some embodiments, in the liquid preparation, at least a part of the vaccine carrier composition forms a vesicle structure, and at least a part of the antigen component is encapsulated in the internal cavity of the vesicle structure or hybridized on the surface of the vesicle structure.

[0186] In some embodiments, when the vaccine preparation is a solid preparation, the solid preparation can be reconstituted to form the aforementioned liquid preparation.

[0187] In some embodiments, the liquid preparation contains vaccine particles. The vaccine particles can be on the nanoscale, and accordingly, the corresponding vaccine preparation is also called a nano-vaccine preparation.

[0188] The average particle size of the vaccine particles can be 30 nm to 1000 nm, can also be 100 nm to 1000 nm, can also be 300 nm to 1000 nm, and further can be 300 nm to 500 nm. The average particle size of the vaccine particles can also be any one of the following particle sizes, or a range composed of any two of the following particle sizes: 30 nm, 50 nm, 60 nm, 80 nm, 100 nm, 120 nm, 130 nm, 140 nm, 150 nm, 160 nm, 180 nm, 200 nm, 250 nm, 300 nm, 250 nm, 400 nm, 450 nm, 500 nm, etc. The average particle size of the vaccine particles can be in the ranges of 50 nm to 500 nm, 100 nm to 500 nm, 50 nm to 400 nm, 100 nm to 400 nm, 120 nm to 400 nm, 130 nm to 400 nm, etc.

[0189] It can be understood that the liquid preparation contains a dispersion liquid. The dispersion liquid is used to provide solvent water. The dispersion liquid includes water, and may or may not include a buffer. Non-limitingly, the buffer can include one or more of the salt components of phosphate buffer solution (PBS), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), sodium citrate (SSC), citric acid, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES).

[0190] In the fourth aspect of the present application, a method for preparing a vaccine preparation is provided, which can be used to prepare the vaccine preparation described in the third aspect of the present application.

[0191] In some embodiments, a method for preparing a vaccine preparation is provided, which includes the following steps:

[0192] The liquid preparation is prepared by any one of the following methods:

[0193] Method 1: The nano-vaccine pharmaceutical composition and the antigen component are used to prepare the vaccine preparation by the thin film hydration method; wherein, the antigen component includes non-nucleic acid antigens;

[0194] Method 2: A vaccine preparation is prepared by microfluidic assembly of the nano-vaccine pharmaceutical composition and the antigen component; wherein, the antigen component includes nucleic acid antigens; and

[0195] Method 3: A vaccine preparation is prepared by pressure homogenization and emulsification of the nano-vaccine pharmaceutical composition and the antigen component.

[0196] This preparation method optionally includes (it may or may not include) the step of drying the liquid preparation into a solid preparation.

[0197] In the fifth aspect of the present application, a lipid derivative targeting dendritic cells is provided, which is the lipid derivative DOPCO targeting dendritic cells described in the first aspect of the present application. Details are not repeated here.

[0198] In the sixth aspect of the present application, there is provided the use of the vaccine carrier composition described in the first aspect of the present application, or the nano-vaccine pharmaceutical composition described in the second aspect of the present application, or the lipid derivative targeting dendritic cells described in the fifth aspect of the present application in relation to targeting dendritic cells, including but not limited to the use in the preparation of a nano-vaccine preparation targeting dendritic cells.

[0199] In the seventh aspect of the present application, there is provided the use of the vaccine carrier composition described in the first aspect of the present application, or the nano-vaccine pharmaceutical composition described in the second aspect of the present application, or the vaccine preparation described in the third aspect of the present application, or the vaccine preparation prepared by the preparation method described in the fourth aspect of the present application in the preparation of a drug for preventing and / or treating at least one of tumors and infectious diseases.

[0200] The aforementioned use can be for preventive or therapeutic purposes, or for non-diagnostic and non-therapeutic purposes. For non-diagnostic and non-therapeutic purposes, non-limiting examples include in vitro research, serving as or preparing a vaccine reference preparation, etc.

[0201] The vaccine carrier composition and the nano-vaccine pharmaceutical composition provided by the present application have the characteristic of targeting dendritic cells. The prepared nano-vaccine is a targeted nano-vaccine capable of targeting dendritic cells, having the ability to induce the production of a wide range of cross-reactive antibodies and cellular immunity, and can target lymph node dendritic cells and be efficiently taken up by dendritic cells, thereby inducing an antigen-specific immune response, and having broad application prospects.

[0202] The targeted nano-vaccine provided by the present application can be used for at least one of the prevention and treatment of diseases such as tumors (such as cancers), infectious diseases, etc. For example, it can be used for tumor immunity, prevention and / or treatment of infections, etc. In addition, the design of the nano-vaccine targeting dendritic cells provided by the present application is of great significance for the development of universal tumor or infectious disease vaccines.

[0203] Non-limitingly, the drug targeting dendritic cells is any one of the drugs for preventing and treating tumors and the drugs for preventing and treating infectious diseases.

[0204] In some embodiments, the tumor may include, but is not limited to, at least one of triple-negative breast cancer, pancreatic cancer, and colorectal cancer.

[0205] In some embodiments, the infectious disease may include, but is not limited to, at least one of novel coronavirus infection, influenza, hepatitis, infectious intestinal disease, pulmonary tuberculosis, neonatal tetanus, mumps, and varicella.

[0206] In the eighth aspect of the present application, a method for preventing or treating a disease is provided, which includes administering a therapeutically effective amount of the aforementioned nano-vaccine pharmaceutical composition or vaccine preparation to a subject.

[0207] As used herein, "therapeutically effective amount" refers to the amount required to cause a biological or medical response in an individual, for example, the dosage that brings about a physiologically and / or pharmacologically positive effect to the individual. "Physiologically and / or pharmacologically positive effect" includes, but is not limited to, reducing or inhibiting enzyme or protein activity, or improving symptoms, alleviating the disease condition, slowing down or delaying the disease process, or preventing the disease, etc.

[0208] As used herein, "subject" refers to an animal, preferably a mammal, and further preferably a human. The term "mammal" mainly refers to warm-blooded vertebrate mammals, including, but not limited to: cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs, cows, sheep, horses, and humans.

[0209] In some embodiments, the administration method is injection.

[0210] In one aspect of the present application, a preparation method of a lipid derivative DOPCO targeting dendritic cells is further provided, which can be prepared from M 1a , M 2a and M 3a as raw materials. Among them, the definitions of R 11 , R 12 , L 11 , L 21 , m, k, and R3 can be referred to the context. R N , F 11 , F 21 are each independently a reactive group or its precursor. Among them, R N can react with F 11 to form a covalent bond, and F 21 or activated F 21 or modified F 21 can react with the amino group in M 3a to form a covalent bond and make M 3aConverted to U 3a The residues shown.

[0211]

[0212] In some embodiments, R N and F 11 are capable of reacting to form an amide bond (-NH-C(=O)-), in which case, R N can be -NH2, and F 11 can be a carboxyl group, an acyl halide, or an activated carboxyl group (such as an active ester group, further such as a succinimidyl carboxylate group).

[0213] Taking the example of the connection between the lipid structural unit and the polyethylene glycol structural unit through an amide bond, M 1a and M 2a can be reacted to form the structure shown by Xa, and then F 21 or activated F 21 or modified F 21 is reacted with the amino group in M 3a to form a covalent bond connection. Taking the example of the connection between the polyethylene glycol structural unit and the chitosan oligosaccharide structural unit through a carbamate group, F 21 or modified F 21 can be an active carbonate group (such as a succinimidyl carbonate group), so as to be capable of reacting with an amino group to form a carbamate group linker. In some embodiments, F 21 can be formed by the coupling reaction between the terminal hydroxyl group of the polyethylene glycol chain and N,N'-disuccinimidyl carbonate (DSC).

[0214]

[0215] The embodiments of the present application will be described in detail below in conjunction with some examples. It should be understood that these examples are only used to illustrate the present application and not to limit the scope of the present application. For the experimental methods without specified conditions in the following examples, the guidance given in the present application is preferably referred to, and it can also be carried out according to the experimental manuals or conventional conditions in the art, or according to the conditions recommended by the manufacturer, or by referring to the experimental methods known in the art.

[0216] In the following examples, for the measurement parameters of the raw material components, if there is no special description, there may be slight deviations within the weighing accuracy range. For the temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0217] I. Preparation of lipid derivative DOPCO targeting dendritic cells

[0218] Example 1. Preparation of lipid derivative DOPCO material targeting dendritic cells (also denoted as DOPCO lipid material)

[0219] Using DOA, polyethylene glycol (PEG) derivatives (structure as shown in M 2c shown) and chitosan oligosaccharide as starting materials, the molecular weight and dosage information of each raw material can be referred to Table 1.

[0220]

[0221] Preparation of DOPCO-1 to DOPCO-4:

[0222] According to Table 1, add the corresponding proportion of DOA to 15 mL of chloroform and stir to dissolve, then add the corresponding proportion of PEG derivative raw material and continue to stir and react for 8 hours; after the reaction is completed, spin-dry with a rotary evaporator. Add N,N'-disuccinimidyl carbonate (DSC) and chloroform to the reaction vessel, mix well, and react overnight at room temperature for 12 hours (to make DSC react with the -OH end of PEG to form succinimidyl carbonate groups). After the room temperature reaction is completed, spin-dry with a rotary evaporator. Then add the corresponding proportion of chitosan oligosaccharide dissolved in DMF, and add a small amount of pure water as a co-solvent, and react overnight at room temperature for 12 hours. Finally, remove excess impurities such as chitosan oligosaccharide by dialysis method, place it in a freeze dryer, quickly cool the solution to below -60 °C and keep it warm for 8 hours to obtain a frozen body, perform vacuum treatment to make the vacuum degree less than 10 Pa, then heat the frozen body to 4 °C - 15 °C and keep it warm for 8 hours, and then heat the frozen body to 32 °C and keep it warm for 8 hours, and introduce inert gas nitrogen, and finally obtain DOPCO freeze-dried powder.

[0223] The structure of the prepared DOPCO product was identified by the following method: Weigh the DOPCO lipid material, dissolve it in deuterated chloroform, and perform hydrogen spectrum characterization on the synthesized material by a nuclear magnetic resonance instrument, and record 1 the 1H NMR spectrum; Weigh DOPCO and dissolve it in 0.5 mL of methanol. Characterize it with a matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometer and record the MALDI-TOF spectrum.

[0224] Table 1. Different molecular weights and dosages of raw materials used to prepare DOPCO

[0225]

[0226] II. Preparation of nano-vaccine

[0227] (I) Preparation of nano-vaccine

[0228] According to the prescription shown in Table 2, the nano-vaccine was prepared by the methods of Examples 2 - 4 respectively.

[0229] Table 2. Composition of nano-vaccine containing DOPCO and content ratio of each component

[0230]

[0231]

[0232] Example 2.

[0233] Preparation of Formulations 2-1, 2-2, 2-3, 2-6, 2-7, and 2-8: Weigh the synthesized DOPCO in the corresponding proportions, add the co-lipid, dissolve it with chloroform, place it in a constant temperature water bath at 60 °C, and dry it under reduced pressure to form a film. Prepare liposomes using the thin film hydration method. After evaporating the chloroform on a rotary evaporator, add the whole cell lysate, cell membrane, or exosomes in the corresponding proportions using injection water as the solvent for hydration (see Table 2), and add the corresponding adjuvants and other pharmaceutical excipients. Finally, ultrasonically crush it with an ultrasonic instrument or homogenize it with a high-pressure homogenizer pump, and then filter and sterilize it with a 0.2 μm filter membrane to prepare a nano-vaccine injection with an antigen active ingredient of 1 mg / mL.

[0234] Example 3.

[0235] Preparation of Formulations 2-4 and 2-5: Weigh the synthesized DOPCO in the corresponding proportions, add the co-lipid, dissolve it with ethanol as the oil phase, dissolve the mRNA adjuvant with acetic acid buffer (pH 4) as the water phase, simultaneously inject the water phase and the oil phase into the microfluidic device, pass through the microfluidic chip at a flow rate of 10 mL / min, and collect the resulting solution. Place it in a 500 Da dialysis bag and dialyze it in PBS for 16 h to remove the organic solvent. After dialysis, add the cell membrane antigen component dispersed in injection water to the formulation, and add other pharmaceutical excipients. Finally, ultrasonically crush it with an ultrasonic instrument or homogenize it with a high-pressure homogenizer pump, then filter and sterilize it with a 0.2 μm filter membrane, place it in a freeze dryer, quickly cool the solution to about -65 °C and keep it warm for 8 h to obtain a frozen body, perform vacuum treatment to make the vacuum degree less than 10 Pa, then warm the frozen body to above 5 °C and keep it warm for 8 h, and then warm the frozen body to 32 °C and keep it warm for 8 h, and introduce the inert gas nitrogen to obtain a nano-vaccine injection with an antigen active ingredient of 1 mg / mL.

[0236] Example 4

[0237] Preparation of Prescription 2-9: Weigh the synthesized DOPCO in the corresponding proportion, add auxiliary lipid, dissolve it with ethanol as the oil phase, dissolve the mRNA antigen with acetic acid buffer solution (pH 4) as the aqueous phase, inject the aqueous phase and the oil phase into the microfluidic device simultaneously, pass through the microfluidic chip at a flow rate of 10 mL / min, and collect the obtained solution. Place it in a 500 Da dialysis bag and dialyze it in PBS for 16 h to remove the organic solvent. After dialysis, add other pharmaceutical excipients to the preparation, finally ultrasonically crush it with an ultrasonic instrument or homogenize it with a high-pressure homogenizer pump, then filter and sterilize it with a 0.2 μm filter membrane, place it in a freeze dryer, quickly cool the solution to about -65 °C and keep it warm for 8 h to obtain a frozen body, conduct vacuum treatment to make the vacuum degree less than 10 Pa, then heat the frozen body to above 5 °C and keep it warm for 8 h, and then heat the frozen body to 32 °C and keep it warm for 8 h, and introduce inert gas nitrogen to obtain a nano-vaccine injection with an antigen active ingredient of 1 mg / mL.

[0238] Example 4.

[0239] Preparation of Prescriptions 3-1 to 3-8: Take the prepared DOPCO lipid material in the corresponding proportion, add auxiliary lipid, dissolve it with chloroform, place it in a constant temperature water bath at 60 °C and dry it into a film under reduced pressure, prepare liposomes by the thin film hydration method, after evaporating chloroform on a rotary evaporator, add injection water and antigen components for hydration, and add other pharmaceutical excipients, finally ultrasonically crush it with an ultrasonic instrument or homogenize it with a high-pressure homogenizer pump, then filter and sterilize it with a 0.2 μm filter membrane to prepare a nano-vaccine injection with an antigen active ingredient of 1 mg / mL.

[0240] Preparation of the control group preparation:

[0241] Take a certain amount of lipid materials DPPC and cholesterol in an eggplant-shaped flask, add 1 mL to 2 mL of chloroform to dissolve, under the condition of 37 °C, remove chloroform through a rotary evaporator, after the lipid forms a uniform thin film, add aqueous phase PBS to hydrate and wash the film to form a relatively uniform suspension, and conduct probe ultrasound treatment at 1% power in an ice bath for 5 min to obtain blank liposomes DPPC-LIPS as the control group.

[0242] (II) Characterization of the nano-vaccine

[0243] The particle size and potential of the nano-vaccine injection were tested by the following method. The results can be referred to Table 3 and Figure 1 . It can be seen that the prepared nano-vaccine has a relatively appropriate particle size and is negatively charged.

[0244] Particle size measurement method: Dilute the sample with deionized water. Select an appropriate particle size cell according to the type of measurement, inject the sample into it, and then place it in the measuring instrument. Measure the particle size of the sample at room temperature. Measure three times for each group, take the average value, and record it as "particle size". After the measurement of one sample is completed, put another sample cell in and repeat the above steps for measurement.

[0245] Zeta potential measurement method: Dilute the sample with deionized water. Select an appropriate zeta potential cell according to the type of measurement, inject the sample into it, and then place it in the measuring instrument. Measure the zeta potential of the sample at room temperature. Measure three times for each group, take the average value, and record it as "zeta potential". After the measurement of one sample is completed, put another sample cell in and repeat the above steps for measurement.

[0246] The test results of particle size and zeta potential can be referred to Table 3.

[0247] Table 3. Particle Size and Zeta Potential of the Formulation

[0248] Prescription Particle size Potential Prescription 2-1 200nm -22mV Prescription 2-2 180nm -30mV Prescription 2-3 175nm -28mV Prescription 2-4 367nm -42mV Prescription 2-5 256nm -28mV Prescription 2-6 181nm -38mV Prescription 2-7 176nm -67mV Prescription 2-8 152nm -19mV Prescription 2-9 139nm -19mV Prescription 2-10 201nm -27mV Prescription 3-1 286nm -0.6mV Prescription 3-2 397nm 0.7mV Prescription 3-3 262nm -31mV Prescription 3-4 345nm -36mV Prescription 3-5 263nm -38mV Prescription 3-6 161nm -51mV Prescription 3-7 291nm -30mV Prescription 3-8 181nm -28mV Prescription 3-9 211nm -33mV Prescription 3-10 145nm -41mV

[0249] (III) Biological Effect Test Method and Result Analysis

[0250] 3.1. Uptake of the Preparation by DC

[0251] To evaluate the uptake of the nano-vaccine by DC, prepare a nano-vaccine labeled with fluorescein DiI. Extract and culture mouse bone marrow-derived dendritic cells (BMDC) until the seventh day, add the DiI-labeled nano-vaccine to BMDC respectively, and replace the cell culture medium. After co-incubating with BMDC for 6 h, discard the supernatant, and wash the cells with PBS. Collect BMDC for flow cytometry analysis. In addition, culture BMDC in a 12-well plate containing cover slips, add the above DiI-labeled preparations of each group and co-incubate for 6 h. After the incubation, wash the cell cover slips with PBS, fix the cells with 4% paraformaldehyde, stain the cell nuclei with DAPI, and observe the fluorescence differences of different groups with a confocal microscope.

[0252] The test results can be referred to Figure 2 and Figure 3 . The nano-vaccine prepared according to the technical solution of the present application has a good DC cell uptake effect, which proves that DC targeting can be generated.

[0253] 3.2. Lysosomal Escape Experiment in DC Cells

[0254] Extract and culture BMDCs, and inoculate the obtained BMDC cells into a 24-well plate at an appropriate density. When the cells grow to 60%-70%, discard the cell culture medium, add the preparation diluted with blank medium, and co-incubate with the cells for 0.5 h and 2 h. After the incubation, remove the preparation, wash the cells, incubate overnight with rabbit anti-LAMP1 primary antibody to label lysosomes, remove the primary antibody the next day, wash and then add Cy3-labeled goat anti-rabbit secondary antibody and incubate for 1.5 h. Finally, perform nuclear staining with DAPI. After making the slides, observe the co-localization of RNA and lysosomes under a laser confocal microscope.

[0255] The test results can be referred to Figure 4 . The nano-vaccine prepared according to the technical solution of the present application can stably encapsulate nucleic acids and enable them to escape from lysosomes, so that the encapsulated nucleic acids can be avoided from being degraded by lysosomal enzymes and further play a role.

[0256] 3.3. DC maturation experiment

[0257] Culture BMDCs for 6 to 8 days, add the prepared nano-vaccine into the culture dish, and co-incubate with BMDCs for 48 h. After the incubation, collect the BMDCs, wash them with PBS, resuspend them in FACS buffer, and label with FITC-CD11c, PE-CD86, and APC-CD80 antibodies according to the instructions. After incubating in the dark for 15 min, centrifuge and collect the labeled BMDCs, wash them, and disperse them in 500 μL of flow cytometry buffer. Detect the proportions of CD80+ and CD86+ in CD11c-positive cells by flow cytometry.

[0258] The test results can be referred to Figure 6 . The nano-vaccine prepared according to the technical solution of the present application can effectively activate the maturation of DC cells and has an immune activation effect.

[0259] 3.4. DC cytokine secretion ability

[0260] Culture BMDCs for 6 to 8 days, add the prepared nano-vaccine into the culture dish, and co-incubate with BMDCs for 48 h. After the incubation, collect the cell supernatant, centrifuge and discard the precipitate. Use the IL-12 Elisa kit and TNF-α Elisa kit from Shanghai Jingkang Biotechnology Co., Ltd. to perform quantitative detection through a multi-functional microplate reader (Varioskan LUX, Thermofisher Scientific) to test the secretion levels of interleukin-12 and tumor necrosis factor.

[0261] The test results can be referred to Figure 5, wherein, "Interleukin-12" in the ordinate represents the secretion level of IL-12, and "Tumor Necrosis Factor" represents the secretion level of TNF-α. The nano-vaccine prepared according to the technical solution of the present application can promote DC to secrete pro-inflammatory cytokines and promote the differentiation and maturation of DC, which is an important process for generating anti-tumor immunity and also the first and crucial step in activating T cells.

[0262] 3.5. Activation ability of T cells

[0263] The extracted BMDCs were cultured for 6 to 8 days, and the prepared nano-vaccines were added to the culture dishes respectively and co-incubated with BMDCs for 48 h. After the incubation, splenic lymphocytes were added to the BMDCs and co-cultured for 48 h. After the culture, the cells were collected, and the activation of T cells co-incubated with DC was examined by flow antibody staining, and the cytokine secretion of T cells was examined by extracting the cell supernatant at the same time.

[0264] The test results can be referred to Figure 7 . The nano-vaccine prepared according to the technical solution of the present application has a significant activating effect on T cells.

[0265] 3.6. Tumor treatment effect

[0266] In the animal experiment, 6-week-old female BALB / c mice were used. After anesthesia, the mice were injected with 4T1 cells (5×10 5 cells per mouse, resuspended in 100 μL PBS), and the tumor volume (V) = 1 / 2 × length × width 2 . Seven days later, the modeled mice were randomly divided into 3 experimental groups (n = 10). When the tumor volume was close to 50 mm 3 , a nano-vaccine with an antigen active ingredient concentration of 10 mg / kg and a volume of 100 μL was given by intramuscular injection into the right thigh every seven days. The tumor volume was measured every 2 - 3 days during the treatment. The treatment ended after four administrations.

[0267] The test results can be referred to Figure 8 . The nano-vaccine prepared according to the technical solution of the present application caused large-area necrosis of tumor cells, verifying the successful destruction of tumor cells by the nano-vaccine and having a significant anti-tumor effect.

[0268] 3.7. Pseudovirus neutralization experiment

[0269] Digest and count the 293T cells in good growth state, then dilute them and add them into a 96-well plate. Place the plate in an incubator at 37°C with 5% carbon dioxide for cultivation. Add the virus on the second day. Make gradient dilutions in an EP tube, aspirate the original culture medium in the 96-well plate, and add the diluted virus solution. Then make good marks. Add additional culture medium on the third day, and add 100 μL of complete culture medium to each well to facilitate cell growth. Observe the results and calculate the titer on the fifth day. Observe the results under a fluorescence microscope, count the number of the last two fluorescent cell clones with fluorescence, and calculate the TCID50 of the virus solution.

[0270] The test results can be referred to Figure 9 . The nano-vaccine prepared according to the technical solution of the present application can effectively neutralize the virus and has antiviral efficacy.

[0271] The technical features of the above embodiments and examples can be combined in any suitable manner. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered to be within the scope described in this specification.

[0272] The above embodiments and examples only represent several embodiments of the present application, which are convenient for understanding the technical solution of the present application in detail, but should not be construed as a limitation on the protection scope of the present application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can still be made, and these all belong to the protection scope of the present application. In addition, it should be understood that after reading the above teachings of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments based on the technical solution provided by the present application are all within the protection scope of the appended claims of the present application. Therefore, the protection scope of the patent of the present application should be subject to the content of the appended claims, and the description and drawings can be used to explain the content of the claims.

Claims

1. A vaccine carrier composition, characterized in that, Each component includes the following mass percentages: 30% to 60% of the lipid derivative DOPCO targeting dendritic cells, 1% to 55% of co-lipids, 0% to 15% of adjuvant components, and 0% to 15% of other pharmaceutical excipients; Among them, the structure of the lipid derivative DOPCO targeting dendritic cells is as follows: Lipid structural unit - L 10 - Polyethylene glycol structural unit - L 20 - Chitosan oligosaccharide structural unit; wherein, L 10 and L 20 are each independently a divalent linking group; The lipid structural unit has the formula U 1a The structure shown: Among them, R 11 and R 12 are each independently a hydrocarbon group having 13 to 17 carbon atoms, and R 11 and R 12 each independently contain 0, 1 or 2 carbon-carbon double bonds; The number-average molecular weight of the polyethylene glycol structural unit is 1000 Da to 8800 Da; The number-average molecular weight of the chitosan oligosaccharide structural unit is 340 Da to 3200 Da.

2. The vaccine vector composition according to claim 1, characterized in that, The lipid derivative DOPCO targeting dendritic cells satisfies one or more of the following characteristics: The lipid structural unit and the polyethylene glycol structural unit are connected by an amide bond; The polyethylene glycol structural unit is connected to the amino site in the chitosan oligosaccharide structural unit through a carbamate group; R 11 and R 12 each independently represents a hydrocarbyl group having 13 to 17 carbon atoms and having one carbon-carbon double bond; The number-average molecular weight of the polyethylene glycol structural unit is 1500 Da to 3000 Da; The number-average molecular weight of the chitosan oligosaccharide structural unit is 100 Da to 1700 Da; The number-average molecular weight of the lipid derivative DOPCO targeting dendritic cells is 1842 Da to 11818 Da; The mass ratio of the lipid structural unit in the lipid derivative targeting dendritic cells is 25% to 55%; The mass ratio of the polyethylene glycol structural unit in the lipid derivative targeting dendritic cells is 30% to 60%; The mass ratio of the chitosan oligosaccharide structural unit in the lipid derivative targeting dendritic cells is 25.33% to 55%.

3. The vaccine vector composition according to claim 1, wherein The lipid derivative DOPCO targeting dendritic cells has a general formula structure shown in Formula Ia: L 11 and L 21 each independently represents an alkylene group having 1 to 4 carbon atoms; m is an integer selected from 22 to 200; COS is a chitosan oligosaccharide structural unit connected through its amino site.

4. The vaccine vector composition according to claim 3, wherein The lipid derivative DOPCO targeting dendritic cells has a general formula structure shown in Formula Id: Among them, k is an integer selected from 1 to 20; R3 is a hydrogen atom or an acetyl group.

5. The vaccine vector composition according to any one of claims 1 to 4, characterized in that, Satisfies one or more of the following characteristics: The vaccine carrier composition includes the following components in mass percentages: 40% to 60% of the lipid derivative DOPCO targeting dendritic cells, 10% to 50% of co-lipids, 2.5% to 15% of adjuvant components, and 4.5% to 15% of other pharmaceutical excipients; The number-average molecular weight of the lipid derivative DOPCO targeting dendritic cells is 3008 Da to 6620 Da; The co-lipids are selected from one or more of the following lipid components: hydrogenated soy phosphatidylcholine, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, PEG-DSPE, and cholesterol; among them, PEG-DSPE is a polyethylene glycolylated derivative of distearoyl phosphatidylethanolamine, and the number-average molecular weight of the PEG part in the PEG-DSPE is 1000 Da to 2000 Da; The adjuvant component is selected from one or more of the following components: Toll-like receptor agonists, Sting agonists, saponin adjuvants, aluminum adjuvants, Freund's adjuvants, poly(ethylene glycol)-poly(lactic-co-glycolic acid) copolymer, and squalene; the Toll-like receptor agonists include at least one of single-stranded mRNA, peptidoglycan, monophosphoryl lipid A, imiquimod, CpG oligonucleotides, bacterial flagellin, polyinosinic:polycytidylic acid, and lipoteichoic acid; the Sting agonists include at least one of CDNs and cGAMP; the saponin adjuvants include at least one of QS-21 and Quil-A; the aluminum adjuvants include at least one of aluminum phosphate, aluminum hydroxide, and potassium alum; the concentration of squalene is 0.5 wt% to 2.5 wt%. The other pharmaceutical excipients are selected from one or more of the following pharmaceutical excipients: surfactants, emulsifiers, sustained-release agents, solubilizers, buffers, osmotic pressure regulators, antibacterial agents, protein stabilizers, vaccine protectants, and fillers.

6. A nano-vaccine pharmaceutical composition, characterized in that, It includes an antigen component and the vaccine carrier composition according to any one of claims 1 to 5.

7. The nano-vaccine pharmaceutical composition according to claim 6, wherein It satisfies one or more of the following characteristics: The mass ratio of the antigen component to the vaccine carrier is 0.1 to 10. The antigen component is a recombinant protein, mRNA, inactivated virus, attenuated virus, or polypeptide. The antigen component is at least one of a tumor-associated antigen and an infectious disease-associated antigen; wherein, the source of the tumor-associated antigen includes at least one of the following: whole cell lysate of tumor cells, tumor cell membrane, tumor cell membrane, and tumor cell exosomes; the infectious disease-associated antigens include at least one of novel coronavirus antigen, influenza virus, hepatitis virus, infectious intestinal diseases, pulmonary tuberculosis, neonatal tetanus, mumps, and varicella.

8. A vaccine preparation, characterized in that, It includes the nano-vaccine pharmaceutical composition according to claim 6 or 7. The vaccine preparation is a solid preparation or a liquid preparation; wherein, the liquid preparation contains vaccine particles, and the average particle size of the vaccine particles is 30 nm to 1000 nm. In the liquid preparation, at least a part of the vaccine carrier composition forms a vesicle structure, and at least a part of the antigen component is encapsulated in the internal cavity of the vesicle structure or hybridized on the surface of the vesicle structure. When the vaccine preparation is a solid preparation, the solid preparation can be reconstituted to form the liquid preparation.

9. The vaccine preparation according to claim 8, wherein It satisfies one or more of the following characteristics: The liquid preparation contains vaccine particles, and the average particle size of the vaccine particles is 100 nm to 500 nm. The liquid preparation contains a dispersion liquid; the dispersion liquid includes water, and may or may not include a buffer; wherein, the buffer includes one or more of the salt components of phosphate buffer solution, tris(hydroxymethyl)methylaminopropanesulfonic acid, sodium citrate, citric acid, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid.

10. A method for preparing the vaccine preparation according to claim 8 or 9, characterized in that, It includes the following steps: The liquid preparation is prepared by using any one of the following methods: Method 1: The nano-vaccine pharmaceutical composition and the antigen component are prepared into the vaccine preparation by the thin film hydration method; wherein, the antigen component includes non-nucleic acid antigens. Method 2: The vaccine preparation is obtained by microfluidic assembly of the nano-vaccine pharmaceutical composition and the antigen component; wherein, the antigen component includes nucleic acid antigens; Method 3: The vaccine preparation is obtained by pressure homogenization and emulsification of the nano-vaccine pharmaceutical composition and the antigen component; Optionally, the preparation method includes a step of drying the liquid preparation into a solid preparation.

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