CN113181351A

CN113181351A - Individual tumor therapeutic vaccine and preparation method thereof

Info

Publication number: CN113181351A
Application number: CN202110466511.2A
Authority: CN
Inventors: 王蒲
Original assignee: Guangzhou Saibaiao Biomedical Technology Co ltd
Current assignee: Guangzhou Saibaiao Biomedical Technology Co ltd
Priority date: 2021-04-28
Filing date: 2021-04-28
Publication date: 2021-07-30

Abstract

The invention relates to the field of biological medicine, and provides an individualized tumor therapeutic vaccine and a preparation method thereof for improving the individualized treatment effect of tumor patients. Individual tumor neoantigen groups are obtained by comparing tumor tissues and normal tissues of a subject, tumor neoantigen peptide groups are synthesized, and the tumor neoantigen peptide groups and adjuvants are used for preparing anti-tumor vaccines, wherein the adjuvants comprise Poly (IC: LC) and STING agonists. The adjuvant consisting of Poly (IC: LC) and STING agonist can effectively activate an organism to generate CD4+ cell immune response and CD8+ cell immune response of a tumor neoantigen peptide group which is expressed abnormally and consists of MHC class I restricted epitope peptide and MHCII restricted epitope peptide aiming at tumor, thereby obviously inhibiting the growth of tumor and generating no obvious adverse reaction.

Description

Individual tumor therapeutic vaccine and preparation method thereof

Technical Field

The invention belongs to the field of biological medicine, in particular relates to an individualized tumor therapeutic vaccine and a preparation method thereof, and particularly relates to application of tumor neoantigen epitope peptide, Poly (IC: LC) and interferon gene stimulating protein STING agonist in tumor resistance.

Background

Malignant tumors are serious diseases threatening human health, and the incidence rate thereof is increased year by year, which is a major challenge facing the current medical research field. In recent years, research on the immunotherapy of tumors has been progressing in a breakthrough. The tumor immunotherapy forms are various and comprise tumor targeting antibodies, immune system regulating antibodies, tumor vaccines, adoptive immune cell therapy and the like.

Therapeutic vaccines have been used for over 200 years since birth, and cannot completely replace drugs at present, but have become an auxiliary means for treating diseases. Therapeutic vaccines aim at inducing antigen-specific T cell responses, capable of killing tumor cells or diseased cells infected with pathogens. The use of antigens to enhance specific immune responses, particularly cellular immune responses, is the direction of development of therapeutic vaccines, with the goal of conferring immunogenicity and immunoreactivity to specific antigens to generate protective cellular immune responses. How to successfully take the specific antigen by the antigen presenting cell and how to break through the low reactivity of the organism to the specific antigen becomes a problem to be solved urgently for a therapeutic vaccine.

The differentiation between tumor cells and normal cells can be described in a wide variety of ways, including proliferative capacity, abnormal expression of multiple different genes, and the like. The most fundamental differences, however, are due to mutations in the tumor cells, which lead to a large increase in the proliferation capacity of the tumor cells, evasion of the immune system monitoring, etc. Mutation is a very essential difference between tumor cells and normal cells, i.e., a product expressed by a mutated gene in tumor cells, such as a peptide chain, is definitely different from that without mutation, while some mutations are presented on the surface of tumor cells through MHC and then are specifically recognized by TCR of T cells, thereby directly killing the tumor cells, and such a mutated peptide segment is called neoantigen ". The new antigen represents an effective target for tumor therapy and is developing into a new method for treating cancer. Because T cells capable of recognizing the new antigens in vivo are rare or tumor cells escape from killing of specific T cells through an immune escape mechanism, the mutant peptide fragments can be synthesized in vitro through a polypeptide synthesis technology and then injected into a human body to stimulate an organism to specifically recognize a large number of T cells of the new antigens to proliferate in vivo, thereby achieving the purpose of specifically killing the tumor cells carrying the new antigens.

Because the immunogen adopted by the tumor therapeutic vaccine is derived from tumor cells, the structure of the vaccine is highly similar to that of wild type protein in normal cells, and the immunogenicity is small; furthermore, to avoid the safety risks associated with the introduction of other immunogenic components, immunogens that are used as tumor therapeutic vaccines are generally small molecules that do not form a complete immunogen to the subject. Furthermore, the therapeutic vaccine is inoculated after the antigen induces the generation of diseases, the inoculation object is a chronic disease patient, the immune response of the chronic disease patient is always in a low level, and the core and difficult problems of the vaccine research are how to break through immune tolerance, enhance the immune response of the organism and improve the effectiveness of the vaccine.

In addition, due to the diversity of tumor pathogenesis and clinical manifestations, even though clinically, tumors belonging to the same type have great difference among different patients, the diversity of tumors and the difference among different patients cause the defect that the treatment effect of the existing therapeutic vaccine is unstable, which is specifically shown in the fact that the tumor therapeutic vaccine has good curative effect in a few individuals, but has poor curative effect or even completely ineffective effect in most cases, and the technical development and application of the tumor therapeutic vaccine are seriously hindered.

In order to solve the problems of poor immunogenicity, unstable effect and the like of a tumor therapeutic vaccine, the prior art mainly searches for a new tumor specific polypeptide with strong immunogenicity, modifies the existing tumor related peptide (for example, by means of connecting a carrier, modifying a structure, fusing expression, molecular simulation and the like), combines and uses a plurality of tumor related peptides, adds an adjuvant and the like. However, the above technical route cannot solve the technical problem at all, but dilutes the effect of the effective immunogen due to the introduction of more immunogens, and introduces more ineffective immunogens and adjuvants, which also brings about higher safety risk.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides an idea of tumor individualized treatment and provides a tumor individualized treatment scheme suitable for industrial popularization.

Provides an individual tumor therapeutic vaccine and a preparation method thereof for improving the individual treatment effect of tumor patients. Individual tumor antigen groups are obtained by comparing tumor tissues and normal tissues of a subject, tumor neoantigen peptide groups are synthesized, and the tumor neoantigen peptide groups and an adjuvant are used for preparing the anti-tumor vaccine, wherein the adjuvant comprises Poly (IC: LC) and STING agonist. The adjuvant consisting of Poly (IC: LC) and STING agonist can effectively activate the organism to generate the immune response of CD4+ cells and CD8+ cells of a tumor neoantigen peptide group which is expressed aiming at tumor abnormality and consists of MHC class I restricted epitope peptide and MHCII restricted epitope peptide, thereby obviously inhibiting the tumor growth and generating no obvious adverse reaction.

In particular, in one aspect, the invention provides a method for preparing a personalized tumor therapeutic composition comprising

(1) Obtaining a set of tumor neoantigen peptides, wherein the set of tumor neoantigen peptides comprises 15-35 polypeptide sequences and each of the tumor neoantigen peptides is an MHC I and MHC II restricted polypeptide 8-35 amino acids in length;

(2) co-preparing a composition of the set of tumor neoantigen peptides of step (1) with an adjuvant comprising Poly IC, Poly LC and STING agonists.

Further, the present invention provides a method for preparing a personalized tumor therapeutic composition, wherein the tumor neoantigen peptide in step (1) is derived from a polypeptide fragment that has a variation in tumor tissue and is capable of binding to an MHC I or MHC II molecule of an individual subject.

Further, the present invention provides a method for preparing a personalized tumor therapeutic composition, wherein the step (1) comprises:

a. carrying out high-throughput sequencing on the tumor tissue and the normal tissue, and analyzing a sequencing result to obtain a polypeptide fragment generated by tumor gene mutation;

b. determining the MHC I and MCH II molecules of the individual subject,

c. obtaining a group of polypeptide fragments with high affinity with MHC I or MHC II molecules of an individual subject as tumor neoantigen peptides through bioinformatics analysis,

d. synthesizing a group of tumor neoantigen peptides obtained in the step c.

Further, the present invention provides a method for preparing a personalized tumor therapeutic composition, wherein in step d, each peptide in the group of tumor neoantigen peptides is chemically synthesized separately and then mixed; the chemical synthesis method comprises a solid-phase synthesis method, a liquid-phase synthesis method and a solid-liquid-phase synthesis method, and the synthesis strategy comprises a C-terminal synthesis method, an N-terminal synthesis method, a sectional synthesis method and the like.

In a second aspect, the present invention also provides a composition obtainable by the process for the preparation of one of the aforementioned personalized tumor therapeutic compositions of the invention.

In a third aspect, the present invention also provides a personalized tumor therapeutic composition comprising

(1) A group of tumor neoantigen peptides, which consists of 15-35 subjects with 8-35 amino acids long MHC I restricted or MHC II restricted tumor antigen polypeptides;

(2) an adjuvant consisting of Poly IC, Poly LC and STING agonist;

(3) a solvent;

and

(4) optionally, pharmaceutically acceptable auxiliary or adjuvant

Further, the present invention provides a personalized therapeutic composition for tumor, wherein the STING agonist is selected from the group consisting of c-AMP-GMP (cgamp), c-di-GMP, c-di-AMP, c-GMP-IMP, and c-di-IMP-c-AMP-GMP.

Further, the individualized tumor therapeutic composition is characterized in that the solvent is an isotonic aqueous solvent, and comprises physiological saline, PBS buffer solution, citric acid buffer solution and the like; the pharmaceutically acceptable auxiliary or adjuvant is preferably DMSO.

Further, the individualized tumor therapeutic composition of the invention is characterized in that each peptide in a group of tumor neoantigen peptides is at a concentration of 100-1000. mu.g/mL, preferably each peptide in a group of tumor neoantigen peptides is at a concentration of 200-800. mu.g/mL, the concentrations of adjuvants Poly IC, Poly LC and STING agonist are respectively 0.5-5mg/mL, 0.5-5mg/mL and 10-500. mu.g/mL, and further preferably the concentrations of adjuvants Poly IC, Poly LC and STING agonist are respectively 1-3mg/mL, 1-3mg/mL and 50-200. mu.g/mL.

In a fourth aspect, the present invention also provides the use of any one of the personalized tumor therapeutic compositions described above in the manufacture of a medicament or vaccine for the treatment of a personalized tumor.

Preferably, the tumor includes, but is not limited to, colon cancer, gastric cancer, ovarian cancer, liver cancer, cervical cancer, melanoma, gastrointestinal tumor, breast cancer, cervical cancer, pancreatic cancer, lung cancer, lymph cancer, and the like.

For a better understanding of the present invention, certain terms are first defined. Other definitions are listed throughout the detailed description section.

The term "vaccine" refers to a biological agent that improves immunity to a particular disease, wherein the vaccine includes an antigen against which an immune response is elicited.

The term "therapeutic vaccine" means that the individual receiving the vaccine already has the disease associated with the antigen of the vaccine, at which point the vaccine can either elicit an immune response or enhance the individual's existing immune response to the antigen to increase the individual's ability to protect against the substance or cell carrying the antigen. This includes immune responses to diseased cells, such as cancer cells, as well as immune responses to disease-associated proteins. Therapeutic vaccines are intended to treat a disease or disorder of interest, such as cancer or an infectious disease, in a subject.

The term "neoantigen" or "neoantigenic (neoantigenic)" means a class of tumor antigens that result from one or more tumor-specific mutations that alter the amino acid sequence of the proteins encoded by the genome.

The term "adjuvant" refers to an agent used to stimulate the immune system and increase the response to a vaccine without having any specific antigenic effect on its own.

The term "Poly IC", polyinosinic cell (Poly I: C), is a copolymer of polyinosinic acid and polycytidylic acid, which is chemically a double stranded RNA (dsRNA) that acts as a TLR3 agonist. The polyinosinic-polycytidylic acid is a ligand of a 3-type Toll-like receptor in vivo, and can mediate a series of immune reactions of an organism after the polyinosinic-polycytidylic acid activates TLR-3, such as the secretion of cytokines such as interferon, interleukin, tumor necrosis factor and the like, the proliferation and maturation of monocytes, macrophages, lymphocytes and dendritic cells are promoted, and the generation of antibodies in vivo is promoted. The polyinosinic-polycytidylic acid has good promotion effect on specific immunity and non-specific immunity of an organism.

The term "Poly LC", i.e. polylysine and carboxymethylcellulose, can be used to stabilize polyinosinic-polycytidylic acid (Poly IC) and extend its half-life. Polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose, which is capable of inducing interferon production in cancer patients, is a TLR3 agonist. TLR3 is expressed in early endosomes of myeloid DCs; polyinosinic acid-polycytidylic acid stabilized by polylysine and carboxymethylcellulose therefore preferentially activates myeloid dendritic cells, thereby contributing to Th1 cytotoxic T cell responses. Poly-ICLC activates Natural Killer (NK) cells, induces cytolytic potential, and induces IFN γ from myeloid DC. The anti-tumor effect of Poly (IC: LC) is to enhance T cell infiltration of solid tumors by MDA-5 and IFN-I mediated means.

The term "STING", an interferon gene stimulating protein, is an important adaptor protein localized to the endoplasmic reticulum membrane that is ubiquitously expressed in mammals. It can sense the DNA of microbe invading host cell, recruit and activate intracellular TANK binding kinase 1 and interferon regulatory factor 3 through dimerization activation of itself, and induce the production of type I interferon and proinflammatory cytokine. STING plays an important role not only in the innate immune response induced by microorganisms, but also in tumor immunity.

The term "STING agonist" refers to any compound that upon contact with STING results in one or more of the following, for example: (1) stimulating or activating STING proteins, (2) enhancing, increasing or promoting, inducing or prolonging STING activity, function or presence and/or (3) enhancing, increasing, promoting or inducing STING expression. Agonist activity can be measured in vitro by various assays known in the art, such as, but not limited to, assays for cell signaling, cell proliferation, markers of immune cell activation, cytokine production. Agonist activity can also be measured in vivo by various assays that measure surrogate endpoints, such as, but not limited to, measuring T cell proliferation or cytokine production, particularly type I interferon.

The term "high throughput sequencing technology" refers to second generation or single molecule sequencing technologies. Second generation sequencing platforms include, but are not limited to, Illumina-Solexa (GATM, HiSeq2000TM, etc.), ABI-Solid, and Roche-454 (pyrosequencing) sequencing platforms; single Molecule sequencing Technologies include, but are not limited to, True Single Molecule sequencing technology (True Single Molecule DNAsequencing) by Helicos, Single Molecule real-time Sequencing (SMRTTM) by Pacific Biosciences, and Nanopore sequencing technology (Rusk, Nicol (2009-04-01) by Oxford Nanopore Technologies, Nature Methods 6(4): 244-245). With the continued evolution of sequencing technologies, those skilled in the art will appreciate that other sequencing methods and devices can also be used to perform whole genome sequencing. According to a specific example of the present invention, the nucleic acid tag according to an embodiment of the present invention may be used for sequencing using at least one selected from Illumina-Solexa, ABI-SOLiD, Roche-454, and single molecule sequencing apparatus.

The term "pharmaceutically acceptable" refers to those approved or approvable by a governmental regulatory agency or for use in animals (including humans) listed in the pharmacopeia or other generally recognized pharmacopeia.

The term "pharmaceutically acceptable adjuvant or adjuvant" includes protective agents, excipients, carriers, diluents, or the like, which may be administered to a subject with an agent, and which do not destroy the pharmacological activity of the agent and are non-toxic when administered in a dose effective to deliver a therapeutic amount of the agent.

The term "treating" means reducing or ameliorating a disorder (e.g., a neoplasia or tumor) and/or a symptom associated therewith. "treating" can refer to administering the combination therapy to a subject after an onset or suspected onset of cancer. "treating" includes the concept of "alleviating," which refers to reducing the frequency of occurrence or recurrence, or reducing the severity of any symptoms or other adverse effects associated with cancer and/or side effects associated with cancer therapy. The term "treating" also encompasses the concept of "managing," which refers to reducing the severity of a particular disease or condition in a patient or delaying the recurrence of the disease or condition, e.g., extending the remission period of a patient with the disease. It will be understood that, although not excluded, treating a disorder or condition does not require complete elimination of the disorder, condition, or symptoms associated therewith.

The term "tumor antigen" refers to a molecule (typically a protein, carbohydrate or lipid) that is expressed, either completely or as a fragment (e.g., MHC/peptide), on the surface of a cancer cell and which can be used to preferentially target a pharmacological agent to a cancer cell. In some embodiments, a "cancer-associated antigen" is a marker expressed by both normal and cancer cells, e.g., carcinoembryonic antigen (CEA). In some embodiments, the cancer-associated antigen is a cell surface molecule that is overexpressed in cancer cells compared to normal cells, e.g., 1-fold overexpressed, 2-fold overexpressed, 3-fold overexpressed, or more compared to normal cells. In some embodiments, a tumor neoantigen is a cell surface molecule that is improperly synthesized in cancer cells, e.g., a molecule containing deletions, additions, or mutations compared to molecules expressed on normal cells. In some embodiments, the tumor neoantigen is expressed exclusively on the cell surface of cancer cells, either completely or as a fragment (e.g., MHC/peptide), and is not synthesized or expressed on the surface of normal cells. Typically, peptides derived from endogenous proteins are presented to the cell surface by Major Histocompatibility Complex (MHC) class I molecules and are recognized by T Cell Receptors (TCRs) on CD8+ T lymphocytes. MHC class I complexes are constitutively expressed by all nucleated cells. In cancer, virus-specific and/or tumor-specific peptide/MHC complexes represent a unique class of cell surface targets for immunotherapy.

The term "subject" or "patient" includes any reference to a human or non-human animal that is the subject of treatment, observation or experiment. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, mice, sheep, dogs, cats, horses, cattle, chickens, etc.

The term "composition" or "pharmaceutical composition" refers to a mixture of a compound of the invention with at least one and optionally more than one other pharmaceutically acceptable chemical component, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and/or excipients.

Compared with the prior art, the technical scheme of the invention has the following beneficial technical effects:

first, a personalized tumor treatment protocol is provided, and a tumor neoantigen (neoantigen) peptide group aiming at an individual subject is obtained through tumor combination of the subject and high-throughput sequencing, MHC affinity prediction and bioinformatics analysis of normal tissues, wherein each antigen polypeptide can stimulate specific anti-tumor immune response in the subject.

Secondly, aiming at the problem of insufficient immunogenicity of the tumor neoantigen (neoantigen), an adjuvant consisting of Poly (IC), Poly (LC) and a STING agonist is provided, and experimental results show that the original sequence of the tumor neoantigen peptide is short and does not have complete immunogenicity.

Thirdly, the novel antigen peptide in the composition is prepared by chemical synthesis, and the immunologic adjuvant and the auxiliary material have definite components, so that the high safety and reliability are ensured while the anti-tumor immune response is effectively stimulated. In addition, the strategy of individually preparing the novel antigen peptide is adopted, so the application range is wide, the influence of cancer types, individual differences and the like is avoided, and the treatment effect has higher stability.

Drawings

Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to refer to like parts throughout the drawings. In the drawings:

FIG. 1: the tumor neoantigen, Poly (IC: LC) and STING excitant are combined to prepare the vaccine to obviously inhibit the growth of the tumor.

FIG. 2: the tumor neoantigen is combined with Poly (IC: LC) and STING excitant to prepare vaccine to obviously enhance the cellular immune response.

FIG. 2A: the tumor neoantigen, Poly (IC: LC) and STING agonist are combined to prepare a vaccine for promoting the IFN-gamma expression of CD4+ cells;

FIG. 2B: the tumor neoantigen, Poly (IC: LC) and STING agonist are combined to prepare a vaccine for promoting the IFN-gamma expression of CD8+ cells;

FIG. 3: the safety of vaccine prepared by combining the tumor neoantigen with Poly (IC: LC) and STING agonist.

From left to right, the cases of liver, spleen, kidney and heart are examined by dissecting.

Detailed Description

Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.

Example 1: identification and synthesis of tumor neoantigen polypeptide

Surgical collection of patient tumor and paracancerous tissue or venous blood samples, whole exon and RNA sequencing of the samples by a second generation sequencer, bioinformatics analysis of mutations using algorithms (e.g., NetMHC, NetMHCpan, MHCFlurry) can be performed using the sequencing data, and the mutations can be caused by missense mutations or insertions/deletions. Translating the mutated gene into polypeptide sequences, and finding out 15-35 polypeptide sequences according to the HLA affinity of a patient; the polypeptides vary in length from 8 to 35 amino acids. The polypeptide is synthesized in a GMP laboratory in a solid phase mode, and the purity of the polypeptide is over 98 percent. Each polypeptide is mixed with 800ug of 200 and 15-35 polypeptides, and 1-2mg of adjuvant is added to prepare the personalized tumor neoantigen vaccine.

Example 2: preparation of novel antigen vaccines

15-35 neo-antigen polypeptides were pooled and divided into 4 polypeptide pools, each of which contained:

5-10 polypeptides with each peptide concentration of 200-

5％DMSO

Physiological saline for injection.

Mixing each peptide pool and an adjuvant to prepare the personalized new antigen polypeptide vaccine, wherein the adjuvant comprises the following components:

1-3mg/ml Poly IC

1-3mg/ml Poly LC

50-200ug/ml STING agonists

Example 3: animal experiments with neoantigen vaccines

The highly tumorigenic mouse Panc02 cell line was cultured in 10% CO in DMEM medium supplemented with 10% fetal bovine serum, 1% L-glutamine and 0.5% penicillin/streptomycin₂37 ℃ is carried out. Whole WES and RNA sequencing of Panc02 cells, identification of immunogenic mutations, and prediction of the affinity of epitope neoantigens to H-2Db and H-2Kb MHC I and MHCII molecules.

Predicted epitopes of 8-35 amino acids in length comprising the identified non-synonymous mutations were analyzed for affinity to H-2Db and H-2Kb MHC I molecules.

Synthesizing polypeptide to prepare new antigen vaccine.

Each group consisted of 8C 57BL/6 mice implanted 10 medial to the hind leg⁶Individual Panc02 cells established a tumor model. The mice were injected with neoantigen vaccine in combination with adjuvant Poly (IC: LC) and STING agonist on

days

0, 3, and 10, respectively, after inoculation of the tumor cells. All mice were kept under pathogen-free conditions and tumor diameters were measured in mice of the experimental and control groups. Experiments prove that the new antigen vaccine can obviously inhibit the growth of tumors (figure 1) and enhance cellular immune response (figure 2). The result shows that the original tumor antigen peptide with a shorter sequence and without complete immunogenicity can effectively stimulate anti-tumor immune response and obviously inhibit tumor growth by being combined with the adjuvant to prepare the anti-tumor vaccine.

The combined application of the novel antigen vaccine and the Poly (IC: LC) and the STING agonist has good safety, the injection amount of the mouse is 100 times of the normal injection amount, and various tissues and organs of the mouse do not show any abnormal changes such as swelling, discoloration and the like (figure 3), and the results show that the Poly (IC: LC) and the STING agonist combined adjuvant not only can enable the organism to generate strong immune response to the incomplete immunogen, but also can not observe toxicity to the liver, spleen, kidney, heart and other solid organs under extreme dosage.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims

1. A method for preparing a personalized therapeutic composition for tumor comprises

2. The method for preparing a personalized tumor therapeutic composition according to claim 1, wherein the tumor neoantigen peptide in step (1) is derived from a polypeptide fragment that has undergone variation in tumor tissue and is capable of binding to MHC I or MHC II molecules of an individual subject.

3. The method of preparing a personalized tumor therapeutic composition according to claim 1, wherein step (1) comprises:

b. determining the MHC I and MCH II molecules of the individual subject,

c. obtaining a group of polypeptide fragments which can be combined with MHC I or MHC II molecules of an individual subject as tumor neoantigen peptides through bioinformatics analysis,

d. synthesizing a group of tumor neoantigen peptides obtained in the step c.

4. A method for preparing a personalized tumor therapeutic composition according to claim 3, wherein each peptide in the set of tumor neoantigen peptides is chemically synthesized separately in step d and then mixed; the chemical synthesis method comprises a solid-phase synthesis method, a liquid-phase synthesis method and a solid-liquid-phase synthesis method, and the synthesis strategy comprises a C-terminal synthesis method, an N-terminal synthesis method, a sectional synthesis method and the like.

5. A composition obtained by the process for the preparation of a personalized tumor therapeutic composition according to any one of claims 1 to 4.

6. A personalized tumor therapeutic composition comprising

(2) an adjuvant consisting of Poly IC, Poly LC and STING agonist;

(3) a solvent;

and

(4) optionally, a pharmaceutically acceptable adjuvant or adjuvant.

7. A personalized therapeutic composition for cancer according to claim 6, characterized in that the STING agonist is selected from the group consisting of c-AMP-GMP (cGAMP), c-di-GMP, c-di-AMP, c-GMP-IMP, c-di-IMP-c-AMP-GMP.

8. The individualized tumor therapeutic composition according to claim 6, wherein said solvent is an isotonic aqueous solvent comprising physiological saline, PBS buffer, citrate buffer, etc.; the pharmaceutically acceptable auxiliary or adjuvant is preferably DMSO.

9. The individualized tumor therapeutic composition according to claim 6, wherein each peptide in the group of tumor neoantigen peptides is present at a concentration of 200-800. mu.g/mL, and the concentrations of the adjuvants Poly IC, Poly LC and STING agonist are present at 1-3mg/mL, 1-3mg/mL and 50-200. mu.g/mL, respectively.

10. Use of a composition according to any one of claims 5 to 9 for the manufacture of a medicament or vaccine for the treatment of a personalized tumor, preferably said tumor includes but is not limited to colon cancer, gastric cancer, ovarian cancer, liver cancer, cervical cancer, melanoma, gastrointestinal tumors, breast cancer, cervical cancer, pancreatic cancer, lung cancer, lymphoid cancer and the like.