MDP nanoparticle and use thereof

Abstract

An MDP nanoparticle and use thereof, relating to the technical field of modular drug therapy. The MDP nanoparticle is a nanoparticle technology for drug conjugation and delivery. By conjugating one drug or a reasonable combination of multiple drugs for disease treatment or prevention onto a high molecular polymer, the MDP nanoparticle can achieve more effective disease treatment or prevention, or act on multiple diseases at the same time, thereby improving the effect of treating or preventing multiple diseases. The present invention also relates to a universal drug conjugation and delivery nanoparticle platform, which enables efficient and precise activation, inhibits or regulates the pathogenesis of one or more diseases, and achieves better preventive and therapeutic effects on diseases either directly or by assisting other drugs, solving the problems of poor targeting property, short half-life, limited administration modes, insufficient safety and inadequate efficacy of various existing drugs, especially for some diseases with complex pathogenesis, such as metabolic diseases, cardiovascular system diseases, and Alzheimer's disease.

Description

MDP nanoparticles and their applications Technical Field

[0001] The present invention relates to the technical field of nanoparticle drug treatment, in particular to an MDP nanoparticle and its application. Background Art

[0002] Currently, various drugs (chemical drugs, peptides, oligonucleotides, proteins, antibodies, cell therapy, polysaccharides, bacteria, viral vectors, gene therapy, etc.) lack cell or tissue targeting, suffer from short half-lives, limited delivery methods (such as oral and respiratory administration), low activation efficiency, and significant side effects. Consequently, a highly effective platform for drug combination therapy and disease prevention is lacking. For example, the adjuvant molecule CpG ODN has the ability to activate the innate immune system by activating the TLR9 receptor. CpG ODNs are categorized into A, B, C, and D classes based on their sequence characteristics and the immune responses they activate. Currently, CpG ODNs are widely used in humans and animals, either alone or in combination with vaccines, immunotherapies, or other drugs and therapeutics, for the prevention and treatment of various diseases, including tumors, infectious diseases, and autoimmune diseases. However, due to their small molecular weight, CpG ODNs are easily cleared from the body, resulting in a short half-life and a lack of cell or tissue targeting. This results in limited efficacy and high side effects. Furthermore, most adjuvants are currently administered via intramuscular or subcutaneous injection. Due to the unique physiological structure and environment of the digestive and respiratory tracts, adjuvant molecules cannot maintain stability and efficacy through oral administration or inhalation. Current solutions include local administration, increasing the dosage and frequency of administration, combining drugs, using different dosage forms, and adopting different delivery methods. However, these approaches have not yet achieved satisfactory results. Similarly, many adjuvant molecules, such as peptides, RNA, DNA, antibodies, proteins, cytokines, polysaccharides, and small molecules, suffer from the same problems of short half-lives, lack of tissue or cell targeting, high side effects, and low efficacy. Furthermore, adjuvants are currently primarily used alone or in simple combinations, resulting in weak effects on immune system activation pathways, an imbalance in the type of immune system activated (Th1 vs. Th2, cellular vs. humoral immunity, IgA vs. IgG), and uncontrollable effects. For complex diseases such as tumors, cardiovascular diseases, respiratory diseases, neurodegenerative diseases, infectious diseases, and metabolic diseases, single-drug treatments have limited efficacy. The trend is towards increasingly combined drug approaches that leverage multiple mechanisms of action to enhance therapeutic or preventive efficacy. However, simply mixing multiple drugs together can be ineffective because they cannot deliver the drugs to the same tissue or cells or effectively activate signaling pathways. For example, simply mixing vaccine antigens with adjuvants is unlikely to achieve the desired disease prevention or treatment effect.

[0003] Currently, problems with the nanoparticle conjugation of various drugs include: 1. Lack of high-throughput, modular ability to combine multiple drugs, 2. Lack of large-scale production, 3. Poor biodegradability, 4. Complex steps for combining drugs and scaffold materials, 5. Low conjugation efficiency, and 6. Poor stability. Summary of the Invention

[0004] To address the aforementioned issues, the present invention provides MDP nanoparticles and their applications. Specifically, the invention relates to a nanoparticle platform for universal drug conjugation and delivery, enabling efficient and precise conjugation and delivery of various drugs, directly or in conjunction with other drugs to achieve enhanced disease prevention and treatment. This addresses current issues with single-drug or combination drug therapy, such as a lack of cell or tissue targeting, short half-lives, limited administration methods (unsuitable for oral or respiratory administration, etc.), low activation efficiency, and significant side effects.

[0005] In a first aspect, the present invention provides an MDP nanoparticle, which is a universal drug conjugation and delivery nanoparticle. By conjugating one or more drugs, such as chemical drugs, CpG1018, MPL, mRNA, siRNA, ASO, PADRE, anti-CD28 antibodies, antibodies that inhibit the immune checkpoint PD1, ADCs, radioisotopes, and covalent or non-covalent conjugates of various drugs, multiple drug molecules and their rational combination on a high molecular weight polymer, multiple pathways are simultaneously modulated to enhance the therapeutic or preventive effects of drugs on diseases, such as the response of innate immune cells including DCs, B cells, NK cells, macrophages, or non-specific activation of immune cells.

[0006] Furthermore, the MDP nanoparticles include: (a) at least one or more drugs, such as CpG ODN sequences of any length; (b) high molecular weight polymers; and or (c) linker molecules, such as Streptavidin, Neutravidin, Rhizavidin and biotin, positively or negatively charged molecules, such as polyamines, polypeptides, polysaccharides, etc.

[0007] Furthermore, the high molecular polymer includes dextran of any length, unmodified or modified glucan of any length, other types of polysaccharides, nucleic acids, polymers containing different high molecules such as PLGA, etc.

[0008] Furthermore, the MDP nanoparticles may be polymers or nanoparticles containing dextran as a component, or polymers other than dextran.

[0009] Furthermore, the MDP nanoparticles include the same drug, such as CpG ODN, or a mixture of two or more different drugs, such as CpG ODN and vaccine antigen.

[0010] Furthermore, the MDP nanoparticles include the same drug, such as CpG ODN, or a mixture of two or more different drugs, such as a mixture of CpG ODN of any length or type and a vaccine antigen.

[0011] Furthermore, each CpG molecule is composed of a natural PE backbone or an artificial PS backbone to increase the stability of the CpG sequence.

[0012] Furthermore, they are conjugated to other molecules and polymeric nanoparticles.

[0013] In a second aspect, the present invention provides an application of the MDP nanoparticles according to any one of the first aspects, the application comprising:

[0014] (a) For the treatment or prevention of diseases in humans or other mammals, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies, metabolic diseases, cardiovascular diseases, CNS diseases, aging, or other diseases;

[0015] (b) For in vitro stimulation of cells, such as dendritic cells, or for in vivo adoptive cell therapy of human or animal diseases;

[0016] (c) alone or in combination with other vaccines or drugs for the treatment or prevention of human or animal diseases, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases;

[0017] (d) As a drug carrier, it can be linked to one or more other drugs, such as antigens, adjuvants, immunostimulatory molecules, cell or tissue targeting molecules, immunomodulators, small molecule drugs, peptides, antibodies, proteins, polysaccharides, nucleic acid drugs (DNA, RNA, siRNA, ASO, etc.), radioisotopes, ADCs, whole or partial cells or pathogens, etc., through any means such as covalent bonds, non-covalent bonds, biotin-avidin, etc., to enhance the prevention or treatment of human or animal diseases;

[0018] (e) For the detection of diseases or in vitro diagnosis of diseases.

[0019] In a third aspect, the present invention provides a pharmaceutical composition comprising the MDP nanoparticles according to any one of the first aspects.

[0020] Furthermore, the pharmaceutical composition also includes pharmaceutically acceptable excipients.

[0021] Furthermore, the pharmaceutical composition is a vaccine composition.

[0022] Furthermore, the pharmaceutical composition is water-soluble or freeze-dried.

[0023] Furthermore, the pharmaceutical composition is used to treat or prevent diseases in humans or other mammals, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases.

[0024] Furthermore, the pharmaceutical composition further comprises component (i): one or more additional biologically active molecules or ingredients, including but not limited to the drugs described in part (d) of the second aspect, such as PD1 or PD-L1 blockers, immunogenic proteins, cell lysates, or all or part of pathogens.

[0025] Furthermore, the administration route of the pharmaceutical composition can be any oral or parenteral route, including but not limited to oral, nasal, intramuscular, intravenous, subcutaneous, intradermal, intraperitoneal injection, etc.

[0026] Furthermore, the pharmaceutical composition can be delivered into the body in any dosage or formulation.

[0027] Furthermore, the pharmaceutical composition can be used to stimulate in vitro cells, such as dendritic cells, for in vivo adoptive cell therapy of human or animal diseases.

[0028] The above technical solution provided by the embodiment of the present invention has at least the following advantages compared with the prior art:

[0029] The present invention provides a modular drug coupling and delivery technology for MDP nanoparticles and its applications. Specifically, the MDP nanoparticles provided by the present invention have the following advantages:

[0030] 1) It has high efficacy, long half-life, and can stimulate multiple receptors simultaneously.

[0031] 2) Safe, biodegradable, and tissue or cell-targeted.

[0032] 3) Multifunctional, able to combine with multiple adjuvants or drug molecules simultaneously.

[0033] 4) The production process is simple, easy to scale up and has good stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0036] FIG1 is a schematic diagram of the structure of a dextran directly bound to a dextran functionalized with an amine group (amino-dextran) in an embodiment of the present invention.

[0037] FIG2 is a schematic structural diagram of an indirect binding of dextran functionalized with an amine group (amino-dextran) in an embodiment of the present invention. Modes for Carrying Out the Invention

[0038] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0039] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0040] In a first aspect, the present invention provides an MDP nanoparticle, which is a universal drug coupling and delivery nanoparticle. One or more drugs, such as CpG1018, MPL, mRNA, siRNA, ASO, PADRE, anti-CD28 antibodies, and antibodies that inhibit the immune checkpoint PD1, are coupled directly or indirectly, covalently or non-covalently, such as with biotin and avidin. Multiple drugs and their rational combinations are attached to a polymer to simultaneously modulate multiple receptors or pathways, thereby improving the drug's efficacy, safety, and half-life, or preventing or treating a wide range of diseases. For example, the MDP nanoparticle can enhance the response of innate immune cells such as DCs, B cells, NK cells, and macrophages, or non-specifically activate immune cells, such as PADRE activating most MHCII subtypes on T helper cells.

[0041] The MDP (MDP stands for Universal Modular Drug Conjugation and Delivery Nanoparticle Technology) provided by this invention relates to a universal modular drug conjugation and delivery nanoparticle platform. For example, TLR9 is a pathogen-associated molecular pattern recognition receptor that recognizes unmethylated CpG motifs (CpG motifs) on viral and bacterial DNA that are not present in human DNA. Its recognition of pathogen DNA triggers and activates signaling pathways in antigen-presenting cells (such as DCs and macrophages), recruiting cytokines and chemokines, thereby regulating innate and adaptive immune responses. Previous studies have shown that TLR9 agonist CpG ODNs have potent immunopotentiating effects against cancer and infectious diseases. TLR9 agonist polymers have also been shown to increase TLR9 accessibility and activate pro-immune responses. The present invention can link multiple natural immunostimulatory molecules, such as oligonucleic acid molecules such as CpG ODN (A, B, C, D, CpG1018, CpG2395, CpG1086, etc.), RNA molecules, siRNA, ASO, polypeptide molecules such as PADRE, polysaccharides, MPL molecules, α-Gal, proteins, anti-CD28, OX40, PD1 antibodies, etc., in a non-covalent (biotin and Neutravidin, Avidin, Streptavidin) or covalent manner; it also includes immune cell targeting molecules (peptides, proteins, polysaccharides, etc.); disease tissue microenvironment regulating molecules (such as tumor microenvironment); to form nanopolymers or nanoparticles.

[0042] In the present invention, MDP is composed of a glucan polymer directly or indirectly coupled to several to thousands of CpG ODN molecules. Compared to CpG monomers, it significantly enhances innate and adaptive immune responses. The advantages of MDP are that the amount of drug required in the organism is low, accompanied by higher safety and efficacy, while also increasing production capacity and reducing production costs.

[0043] MDP nanoparticles can be used alone or in combination with other pharmaceutical ingredients to treat and prevent human or animal diseases.

[0044] As an embodiment of the present application, the MDP nanoparticles include: (a) at least one drug, such as a CpG ODN sequence of any length; (b) a high molecular weight polymer; and or (c) a linker molecule, such as SA and biotin.

[0045] In the present invention, (a) any drug for treating or preventing a disease, such as a CpG ODN sequence, acts by binding to the TLR9 receptor of antigen-presenting cells, activating the innate immune system, and ultimately activating the acquired immune system, directly or adjuvanting the preventive and therapeutic effects of vaccines or drugs on the disease, specifically CpG1018, CpG2395, CpG2006, etc.

[0046] In the present invention, the function of (b) the high molecular weight polymer is to provide a scaffold and shape for binding the drug component, and specifically, it can be dextran, glucan, other polysaccharide molecules of any length, DNA or RNA nucleic acid, polymer PLGA of different high molecular weight materials, etc.

[0047] In the present invention, the function of the (c) linker molecule is to link the drug molecule with the high molecular weight polymer, and specifically, SA and biotin can be used.

[0048] For example, as shown in FIG1-2 , it can be seen from FIG1-2 that: 5'-end biotinylated A, B, C type CpG ODN is combined with a biotinylated 500K molecular weight dextran molecule (Dextran) through a linker Streptavidin to form MDP nanoparticles.

[0049] In some embodiments, the high molecular weight polymer comprises dextran of any length.

[0050] In some specific embodiments, the MDP nanoparticles can be polymers or nanoparticles containing dextran as a component, or polymers other than dextran.

[0051] In some embodiments, the MDP nanoparticles include the same CpG ODN or a mixture of two or more different CpG ODNs.

[0052] In some embodiments, the MDP nanoparticles include the same CpG ODN or a mixture of two or more different CpG ODNs, of any length or type.

[0053] In some embodiments, each CpG molecule is composed of a natural PE backbone or an artificial PS backbone to increase the stability of the CpG sequence.

[0054] In some embodiments, they are bound to other molecules and polymeric nanoparticles. Specifically, "they are bound to other molecules and polymeric nanoparticles" refers to other polysaccharide polymer materials, single-stranded or double-stranded nucleic acids, polypeptides, and derivatives or modified materials of these materials.

[0055] In some specific embodiments, the particle size of the MDP nanoparticles is 1 to 100,000 nm, preferably 50 to 100 nm.

[0056] In some specific embodiments, the preparation method of the above-mentioned MDP nanoparticles includes the following steps: 1. mixing a biotinylated drug molecule such as CpG ODN, a linker such as Streptavidin and a biotinylated dextran polymer material in a certain molar ratio, and incubating at room temperature, 37 degrees, or 4 degrees for 30 minutes to 24 hours; 2. purifying to remove unbound monomer molecules or incompletely bound nanoparticles; 3. ultrafiltration and concentration.

[0057] In a second aspect, based on the same inventive concept, the present invention provides an application of the MDP nanoparticles according to any one of the first aspects, the application comprising:

[0058] (a) for the treatment or prevention of diseases in humans or other mammals, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases;

[0059] (b) For in vitro stimulation of cells, such as dendritic cells, or for in vivo adoptive cell therapy of human or animal diseases;

[0060] (c) alone or in combination with other vaccines or drugs for the treatment or prevention of human or animal diseases, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases;

[0061] (d) As a drug carrier, it can be linked to one or more other drugs, such as adjuvants, immunostimulatory molecules, cell or tissue targeting molecules, immunomodulators, small molecule drugs, peptides, antibodies, various antigens, pMHC complexes, proteins, polysaccharides, nucleic acid drugs (DNA, RNA, siRNA, ASO, etc.), radioisotopes, ADCs, whole or partial cells or pathogens, or covalent or non-covalent combinations of multiple types of drugs, through any means such as covalent bonds, non-covalent bonds, biotin-avidin, etc., to enhance the prevention or treatment of human or animal diseases;

[0062] (e) For the detection of diseases or in vitro diagnosis of diseases.

[0063] In the present invention, the "application" includes the use for preparing products in the above-mentioned application fields, such as drugs, detection kits, detection equipment, etc.

[0064] In a third aspect, based on the same inventive concept, the present invention provides a pharmaceutical composition comprising the MDP nanoparticles described in any one of the first aspects.

[0065] In some specific embodiments, the pharmaceutical composition further includes pharmaceutically acceptable excipients. Such excipients may include "drug carriers" (specifically, systems that can change the way a drug enters the human body and its distribution within the body, control the rate of drug release, and deliver the drug to the targeted organ), "excipients" (such as physiological saline, glucose, vitamin C, amino acids, etc., which refer to additives other than the main drug in a pharmaceutical preparation. Examples include adhesives, fillers, disintegrants, and lubricants in tablets; wine, vinegar, and medicinal juice in traditional Chinese medicine pills; the matrix component of semi-solid ointments and creams; and preservatives, antioxidants, flavoring agents, fragrances, cosolvents, emulsifiers, solubilizers, osmotic pressure regulators, and colorants in liquid preparations.).

[0066] In some specific embodiments, the pharmaceutical composition is a vaccine composition.

[0067] In some embodiments, the pharmaceutical composition is aqueous or lyophilized.

[0068] In some embodiments, the pharmaceutical composition is used to treat or prevent diseases in humans or other mammals, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases.

[0069] In some specific embodiments, the administration route of the pharmaceutical composition can be any oral or parenteral route, including but not limited to oral, nasal, intramuscular, intravenous, subcutaneous, intradermal, intraperitoneal injection, etc.

[0070] In some specific embodiments, the pharmaceutical composition can be delivered into the body at any dose or formulation. Preferably, it is delivered into the body at an effective dose or formulation. "Effective amount" may also be referred to as "therapeutically effective amount", which refers to the amount of the agent sufficient to provide the desired biological result. The result may be a reduction and / or alleviation of the signs, symptoms or causes of the disease, or any other desired change in the biological system. For example, an "effective amount" for therapeutic use refers to the amount of the composition comprising the pharmaceutical composition of the present invention required to clinically significantly reduce the disease. In any individual case, the appropriate "effective" amount can be determined by a person of ordinary skill in the art using routine experiments. Therefore, the expression "effective amount" generally refers to the amount of the active substance when it has a therapeutic effect.

[0071] In some embodiments, the pharmaceutical composition can be used to stimulate in vitro cells, such as dendritic cells, for in vivo adoptive cell therapy of human or animal diseases.

[0072] In some specific embodiments, the pharmaceutical composition further comprises component (i): one or more additional biologically active molecules or ingredients, including but not limited to immunomodulators, such as PD1 or PD-L1 blockers, immunogenic proteins, peptides, cell lysates, or all or part of a pathogen.

[0073] In summary, the present invention provides an MDP nanoparticle and its application. Specifically, the MDP nanoparticle provided by the present invention has the following advantages:

[0074] 1) The drug is highly effective in treating or preventing diseases, has a long half-life, and can stimulate multiple receptors simultaneously.

[0075] 2) Safe, biodegradable, and tissue or cell-targeted.

[0076] 3) Multifunctional, able to combine with multiple adjuvants or drug molecules simultaneously.

[0077] 4) The production process is simple, easy to scale up and has good stability.

[0078] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples where specific conditions are not specified are generally measured in accordance with national standards. If there are no corresponding national standards, then the methods are carried out in accordance with general international standards, conventional conditions, or the conditions recommended by the manufacturer.

[0079] Example 1

[0080] This example provides an MDP nanoparticle, which includes: Biotin-CpG1018, Streptavidin, and Biotin-Dextran 500kd.

[0081] The preparation method of the above-mentioned MDP nanoparticles includes the following steps:

[0082] 1. Mix Biotin-CpG1018 and Streptavidin at a molar ratio of 1:0.1-10:1 and incubate at room temperature for 30 minutes to 24 hours.

[0083] 2. Mix Biotin-Dextran 500kd with the solution obtained in step 1 at a molar ratio of Dextran 500:Streptavidin of 1:1–1:1000 and incubate at room temperature for 30 minutes to 24 hours.

[0084] 3. Chromatographic purification.

[0085] 4. Ultrafiltration and concentration.

[0086] Example 2

[0087] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the linker is Neutravidin; the remaining steps and parameters are the same.

[0088] Example 3

[0089] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the linker is Rhizavidin; the remaining steps and parameters are the same.

[0090] Example 4

[0091] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is Dextran 2000Kd; the remaining steps and parameters are the same.

[0092] Example 5

[0093] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is Dextran 200Kd; the remaining steps and parameters are the same.

[0094] Example 6

[0095] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is Dextran 100Kd; the remaining steps and parameters are the same.

[0096] Example 7

[0097] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is Dextran 70Kd; the remaining steps and parameters are the same.

[0098] Example 8

[0099] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is Dextran 10Kd; the remaining steps and parameters are the same.

[0100] Example 9

[0101] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the polymer material is mannose; the remaining steps and parameters are the same.

[0102] Example 10

[0103] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG2395; the remaining steps and parameters are the same.

[0104] Example 11

[0105] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecules are CpG2395 and CpG1018; the remaining steps and parameters are the same.

[0106] Example 12

[0107] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecules are CpG2395 and PADRE; the remaining steps and parameters are the same.

[0108] Example 13

[0109] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecules are CpG2395 and PADRE; and there is an additional DC cell targeting molecule polypeptide VSY, and the remaining steps and parameters are the same.

[0110] Example 14

[0111] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecules are CpG2395 and PADRE; and there is an additional DC cell targeting molecule polysaccharide Mannose, and the remaining steps and parameters are the same.

[0112] Example 15

[0113] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and the vaccine antigen is influenza HA, and the remaining steps and parameters are the same.

[0114] Example 16

[0115] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and contains a vaccine antigen such as the tumor-associated antigen HER2, and the remaining steps and parameters are the same.

[0116] Example 17

[0117] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and the vaccine antigen is a tumor neoantigen KRAS G12D. The remaining steps and parameters are the same.

[0118] Example 18

[0119] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018; and it contains vaccine antigens such as tumor neoantigen KRAS G12D and PDL1 antigen. The remaining steps and parameters are the same.

[0120] Example 19

[0121] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018; and it contains vaccine antigens such as tumor neoantigen KRAS G12D and IDO protein antigen. The remaining steps and parameters are the same.

[0122] Example 20

[0123] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and the vaccine antigen is a beta-amyloid protein, an Alzheimer's antigen. The remaining steps and parameters are the same.

[0124] Example 21

[0125] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and contains a vaccine antigen such as the hyperlipidemia antigen Lipoprotein a protein. The remaining steps and parameters are the same.

[0126] Example 22

[0127] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018; and it contains anti-CD19 monoclonal antibody and cytotoxic drug Topoisomerase Inhibitor. The remaining steps and parameters are the same.

[0128] Example 23

[0129] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and an anti-TNFR monoclonal antibody is present, and the remaining steps and parameters are the same.

[0130] Example 24

[0131] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and an anti-KRAS G12D ASO nucleic acid drug is present, and the remaining steps and parameters are the same.

[0132] Example 25

[0133] This example provides an MDP nanoparticle and a preparation method thereof, which differs from Example 1 only in that the adjuvant molecule is CpG1018 and the antigen is an influenza HA mRNA sequence. The remaining steps and parameters are the same.

[0134] Test Case

[0135] In this example, the MDP nanoparticles obtained in Examples 1 to 23 were tested.

[0136] 1) DNA electrophoresis test: The test results show that the CpG1018 molecular band is located near the loading hole of the gel.

[0137] 2) Heat denaturation SDS-PAGE test: the test results show that the Neutravidin protein band is around 60kd.

[0138] 3) Heat denaturation SDS-PAGE test: The test results showed that the Rhizavidin protein band was around 30 kd.

[0139] 12) Heat denaturing SDS-PAGE test showed that the PADRE peptide band was around 2 kd.

[0140] 13) Heat denaturing SDS-PAGE test showed that the PADRE and VSY peptide bands were around 1–3 kd.

[0141] 15) Heat denaturation SDS-PAGE test: The test results show that the HA protein band is around 60kd.

[0142] 18) Heat denaturation SDS-PAGE test, the test results show that KRAS G12D and PDL1 protein bands are at 20kd

[0143] and about 30kd.

[0144] 25) Heat denatured DNA gel test: The test results show that the mRNA nucleic acid molecule band is near the gel loading hole.

[0145] In summary, the present invention relates to MDP nanoparticles and their applications. MDP nanoparticles are used as a nanoparticle technology for drug conjugation and delivery. By conjugating one or more therapeutic or preventive drugs, or by rationally combining one or more drugs onto a polymer, these nanoparticles achieve more efficient disease treatment or prevention, or simultaneously act on multiple diseases, enhancing their effectiveness. This invention also relates to a universal drug conjugation and delivery nanoparticle platform that efficiently and precisely activates, inhibits, or modulates the pathogenesis of one or more diseases, directly or in conjunction with other drugs to achieve better disease prevention and treatment. This addresses the challenges of current drugs (small molecules, radiotherapy, proteins, antibodies, mRNA, cell therapy, oncolytic viruses, bacterial drugs, etc.) with poor targeting, short half-lives, limited delivery methods, and insufficient safety and efficacy. This is particularly true for diseases with complex pathogenesis, such as tumors, autoimmune diseases, infectious diseases, metabolic diseases such as obesity and diabetes, cardiovascular diseases, Alzheimer's disease, and aging.

[0146] Various embodiments of the present invention may be presented in the form of a range; it should be understood that the description in a range format is only for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention; therefore, the range description should be considered to have specifically disclosed all possible subranges and single numerical values ​​within the range. For example, the description of a range from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. In addition, whenever a numerical range is indicated herein, it is intended to include any cited numeral (fractional or integer) within the indicated range.

[0147] The foregoing description is intended only to provide specific embodiments of the present invention, which will enable those skilled in the art to understand and implement the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not intended to be limited to the embodiments shown herein, but is intended to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A MDP nanoparticle, characterized in that: The MDP nanoparticle is a nanoparticle that can treat or prevent diseases in humans or other mammals. It can improve the targeting, half-life, diversity of administration methods, safety and efficacy of drugs by modularly and rationally coupling and delivering one or more drugs, such as molecules that can activate the natural immune system or non-antigen specific molecules, multiple immune activation molecules and reasonable combinations on high molecular polymers, while regulating multiple receptors or disease pathways.

2. The MDP nanoparticles according to claim 1, characterized in that The MDP nanoparticles include: (a) at least one drug, such as a CpG ODN sequence of any length; (b) a high molecular weight polymer of any length; and or (c) one or more covalently or non-covalently linked molecules; Wherein, the high molecular polymer includes dextran of any length; the connecting molecule includes Neutravidin, Streptavidin, Avidin, or Rhizavidin and biotin.

3. The MDP nanoparticle according to any one of claims 1 to 2, characterized in that: The MDP nanoparticles include the same drug, such as CpG ODN, or two or more different drugs, such as a mixture of CpG ODN.

4. The MDP nanoparticles according to any one of claims 1 to 3, characterized in that: The MDP nanoparticles include the same drug, such as CpG ODN, or a mixture of two or more different drugs, such as CpG ODN, with any length or type of CpG ODN.

5. The MDP nanoparticles according to claims 1 to 4, characterized in that: Each CpG molecule is composed of a natural PE backbone or an artificial PS backbone to increase the stability of the CpG sequence.

6. The MDP nanoparticle according to any one of claims 1 to 5, characterized in that: They are bound to other molecules and polymeric nanoparticles.

7. The use of the MDP nanoparticles according to any one of claims 1 to 6, characterized in that: The applications include: (a) for the treatment or prevention of diseases in humans or other mammals, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies, metabolic diseases, aging, cardiovascular diseases, CNS diseases or other diseases; (b) for in vitro stimulation of in vitro cells, such as dendritic cells, or for in vivo adoptive cell therapy of human or animal diseases; (c) alone or in combination with other vaccines or drugs for the treatment or prevention of human or animal diseases, including but not limited to cancer, infectious diseases, autoimmune diseases, allergies or other immune-mediated diseases; (d) As a drug carrier, it is linked to one or more other drugs, such as antigens, adjuvants, immunostimulatory molecules, cell or tissue targeting molecules, immunomodulators, small molecule drugs, peptides, antibodies, proteins, polysaccharides, nucleic acid drugs (DNA, RNA, siRNA, ASO, etc.), radioisotopes, ADC, CAR-T, oncolytic viruses, bacterial vector drugs, etc., or covalent or non-covalent combinations of different drugs, etc., to enhance the prevention or treatment of human or animal diseases by drugs; (e) For the detection of diseases or in vitro diagnosis of diseases.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the MDP nanoparticles according to any one of claims 1 to 7.

Images