CN114432439A - Anti-tumor composition and application thereof - Google Patents
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- CN114432439A CN114432439A CN202111487574.2A CN202111487574A CN114432439A CN 114432439 A CN114432439 A CN 114432439A CN 202111487574 A CN202111487574 A CN 202111487574A CN 114432439 A CN114432439 A CN 114432439A
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Abstract
The invention relates to the technical field of medicines, and particularly relates to an anti-tumor composition, application thereof and an anti-tumor medicine. The anti-tumor composition comprises a PD-1 antagonist and a child coprophila capsule. Experiments show that the PD-1 antagonist with different action mechanisms and action ways is combined with the children coprophila capsule, so that the immunity of the organism can be synergistically improved, the curative effect on cancers expressing PD-I (such as liver cancer) is obvious, and the effect is obviously superior to that of single use.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to an anti-tumor composition and application thereof.
Background
Fecal transplantation (FMT), also known as intestinal microbial transplantation and fecal therapy, is the transplantation of fecal bacteria by various means of transplanting fecal flora from healthy donors into the digestive tract of patients in order to reconstruct intestinal flora of patients and thus achieve the purpose of treating intestinal tract internal and external diseases. FMT is currently recommended by clinical medical guidelines and consensus for the treatment of recurrent or refractory Clostridium Difficile Infection (CDI) and is increasingly being used for the treatment of other intra-and extra-intestinal diseases. However, the establishment of FMT methodology is complex, and no unified standard exists at home and abroad, so that the heterogeneity of various research curative effects is large, and the clinical popularization and application of FMT are greatly limited.
The intestinal tract of children contains many of the same flora as adults, but age-related flora varies in classification and function. The intestinal flora of pre-pubertal children is enriched compared to adults with bifidobacteria, faecalis (Faecalibacterium) and the family pilospiraceae (Lachnospiraceae), which are involved in vitamin synthesis, degradation of amino acids, acidification of phosphorus oxide and triggering of mucosal inflammation, and with significant differences in the relative abundance of these flora. In addition, the intestinal flora structure of children is influenced by geography and dietary culture, researches show that the intestinal flora of children in developed countries is rich in bacteroides and firmicutes, and the intestinal tracts of children in developing countries mainly comprise Prevotella. Bacterial flora structures mainly of the genus bacteroides and bifidobacterium are very common in east asia, while bacterial flora structures mainly of the genus prevotella dominate in central asia and southeast asia.
At present, in the prior art, coprinus comatus capsules are mainly used for treating gastrointestinal diseases, and the coprinus comatus capsules are not reported to be used for treating tumors, and the coprinus comatus capsules and PD-1 antagonists are not reported to be combined for treating tumors.
Disclosure of Invention
In view of the above, the present invention provides an anti-tumor composition and an application thereof. The PD-1 antagonist and the coprinus comatus capsule are combined for treating the liver cancer, and the synergistic and enhanced inhibition effect on the growth of the tumor volume of the liver cancer is found.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an anti-tumor composition, which comprises a PD-1 antagonist and a children coprophila capsule; the children fecal strain capsule is prepared from feces of healthy children of 7-16 years old.
In some embodiments, the PD-1 antagonist is in mg by mass and the pediatric fecal bacteria capsule is in CFU by colony count, and the ratio of the mass of the PD-1 antagonist to the colony count of the pediatric fecal bacteria capsule is (0.09-0.1): (2 x 10)6～2×108)。
In some embodiments, the ratio of the mass of the PD-1 antagonist to the colony count of the pediatric fecal bacteria capsule is 0.1: 2X 10 in mg/CFU8。
In some embodiments, the coprophila bassiana capsule is prepared by the following method:
2, screening the qualified donors by utilizing a high-throughput sequencing method;
and 5, mixing the fecal bacteria liquid prepared in the step 4 with a freeze-drying protective agent, cooling, freezing and vacuum-drying to obtain fecal bacteria freeze-dried powder, and filling the obtained fecal bacteria freeze-dried powder into a capsule shell to obtain the children fecal bacteria capsule.
In some embodiments, in step 1, the health questionnaire comprises: gastrointestinal discomfort does not occur in nearly 2 weeks, antibiotics, acid suppressants, immunosuppressive agents, chemotherapeutic drugs and the like are not used in nearly 3 months, no chronic pain symptom, no digestive system operation history, no infectious disease history and infectious disease contact history, no allergic disease, autoimmune disease, metabolic disease, cardiovascular and cerebrovascular diseases, nervous system or psychiatric disease history, no malignant tumor history and no intravenous injection of growth hormone, insulin, blood coagulation factors and the like are received; the medicine has the advantages of regular work and rest, healthy diet, harmony at home, no addiction to smoking, drinking or drug addiction and the like, and no vaccination or medicine test, no tattoo acceptance or skin damage, no contact with affected areas and tropical areas in nearly 6 months; no family history of gastrointestinal lesions, no family history of malignant tumors and no family history of infectious diseases; the psychologist or psychological consultant interview believes that the patient is currently in good mental state; normal scores were obtained on the depression self-rating scale (SDS), the anxiety self-rating scale (SAS), and the Pittsburgh Sleep Quality Index (PSQI).
In the invention, whether the donor contains pathogenic bacteria is identified by adopting high-throughput sequencing, the donor without pathogenic bacteria is screened out, the donor with potential risk is eliminated, and the quality monitoring is facilitated.
In some embodiments, the method for screening donors using high throughput sequencing comprises the steps of:
(1) performing second-generation sequencing on the DNA and/or RNA extracted from the donor to obtain original sequencing data;
(2) removing host genes of the original sequencing data, comparing the host genes with a microbial database, and performing strain identification and abundance detection;
(3) comparing with a pathogen database to confirm that no pathogen exists in the donor;
preferably, the microbial database of step (2) comprises any one of or a combination of at least two of a bacterial genome, a fungal genome or a viral genome derived from a public database.
Preferably, the pathogen database of step (3) comprises pathogen genomes derived from a public database.
According to the invention, public databases such as NCBI, KEGG and the like are utilized to construct a microorganism database and a pathogenic bacteria database, sequencing data of a fecal bacteria donor is compared to the databases, microbial identification at a strain level is realized, the fecal bacteria donor carrying pathogenic bacteria is eliminated, and health risks are reduced.
In the invention, the fecal bacteria donor is screened by using high-throughput sequencing according to the biomarkers expressed by the fecal bacteria donor and the diversity index of the biomarkers in the step 2.
Preferably, the biomarker includes any one of Escherichia coli (Escherichia coli), Clostridium polybrevum (Clostridium ramosus), Eubacterium cylindroides (Eubacterium cylindroides), human Roseburia (Roseburia hominis), Clostridium pratensis (faecalibacterium prausnitzii), Bacteroides fragilis (Bacteroides fragilis) or Bacteroides vulgatus (Bacteroides vulgatus) or a combination of at least two thereof.
Preferably, the diversity index of the biomarker comprises the alpha diversity index of the biomarker.
In the invention, the high-throughput sequencing is combined with a machine learning model to screen out biomarkers of Escherichia coli (Escherichia coli), Clostridium polybotrys (Clostridium ramosum), Eubacterium cylindracea (Eubacterium cylindroids), human Ralstonia rosenbergii (Roseburia hominis), Clostridium pratensis (Faecalibacterium prausnitzii), Bacteroides fragilis (Bacteroides fragilis) and Bacteroides vulgatus (Bacteroides vulgatus) and alpha diversity index of the biomarkers as an index for screening the fecal strain donor.
Preferably, in the step 3, the clinical examination includes: the medical surgery physical examination is negative; a Body Mass Index (BMI) of 18.5-23.9 kg/m 2; normal blood, liver and kidney function, electrolytes and C-reactive protein; hepatitis E-P-butyl hepatitis, HIV1 and HIV2 antibodies, HTLV1 and HTLV2 antibodies, treponema pallidum antibodies (TPHA, VDRL), EBV lgM and lgG, cytomegalovirus lgM and lgG, strongyloides stercorallum serological examination negative; the routine inspection of the excrement is normal; negative in occult blood test; detection negatives of clostridium difficile, campylobacter, vibrio, salmonella, shigella, shiga toxin-producing escherichia coli, yersinia and plesiomonas; the egg, vesicle, parasite, microsporidian, giardia and cryptosporidium fecal antigen, cyclosporins and heterophyllies are acid-resistant staining detection negative; negative detection of norovirus, rotavirus, adenovirus and enterovirus; multiple drug-resistant bacteria: carbapenem-resistant enterobacteriaceae (CRE), broad-spectrum beta-lactamase bacteria (ESBL), methicillin-resistant staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE) test negative; helicobacter pylori fecal antigen detection is negative; negative for serum monogenic hereditary diseases; fecal neocoronavirus (COVID-19) negative.
And after the target donor is screened out, soaking excrement of the target donor in sterile normal saline, filtering, centrifuging, taking a precipitate, and mixing the precipitate with the sterile normal saline to obtain an excrement liquid.
In some embodiments, the temperature of the sterile physiological saline is 3-5 ℃, specifically 3 ℃, 4 ℃ or 5 ℃.
In some embodiments, the filtering is performed using a screen. Wherein, the aperture of the filter screen is 0.25-2 mm, and specifically can be 0.25mm, 0.5mm, 1.0mm or 2.0 mm.
In some embodiments, the filtering comprises removing large particles by using 2.0mm, 1.0mm, 0.5mm and 0.25mm sieves in sequence, and then filtering for 2-3 times by using 0.25mm sieves, so as to obtain the liquid phase of the fecal filtrate.
And (3) after obtaining the fecal filtrate, centrifuging the fecal filtrate, taking the precipitate, and mixing the precipitate with sterile normal saline to obtain fecal bacteria liquid.
In some embodiments, the rotational speed of the centrifugation treatment is 1500-3000 r/min, specifically 1500r/min, 1600r/min, 1700r/min, 1800r/min, 1900r/min, 2000r/min, 2100r/min, 2200r/min, 2300r/min, 2400r/min, 2500r/min, 2600r/min, 2700r/min, 2800r/min, 2900r/min or 3000 r/min. The time of the centrifugal treatment is 10-20 min, specifically 10min, 11min, 12min, 13min, 14min, 15min, 16min, 17min, 18min, 19min or 20 min.
In some embodiments, the lyoprotectant includes skim milk powder, trehalose, sucrose, vitamin C, and sterile saline. In some embodiments, the lyoprotectant comprises, in mass percent: 10-20% of skim milk powder, 10-15% of trehalose, 1-10% of sucrose, 1-5% of vitamin C and the balance of normal saline.
According to the invention, the freeze-drying protective agent can effectively prolong the survival time of the flora and improve the planting effect of the flora.
The volume ratio of the fecal bacteria liquid to the freeze-drying protective agent is (2-5) to 1, specifically 2:1, 3:1, 4:1 or 5:1, and preferably 3: 1.
Preferably, the temperature reduction and freezing conditions are that the temperature is reduced from room temperature to 3-6 ℃ in 10-20 s, the temperature is reduced from 3-6 ℃ to-30-50 ℃ at 1-2 ℃/min, and the temperature is reduced from-30 ℃ to-50 ℃ at 4-5 ℃/min to-75-80 ℃.
In some embodiments, the time for cooling and freezing is 12-24 h, and specifically may be 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h or 24 h.
In some embodiments, the vacuum degree of the vacuum drying is 5 to 15pa, for example, 5pa, 6pa, 7pa, 8pa, 9pa, 10pa, 11pa, 12pa, 13pa, 14pa or 15 pa.
In some embodiments, the temperature of the vacuum drying is from-50 to-60 ℃, specifically can be-50 ℃, -51 ℃, -52 ℃, -53 ℃, -54 ℃, -55 ℃, -56 ℃, -57 ℃, -58 ℃, -59 ℃, or-60 ℃.
In some embodiments, the vacuum drying time is 24-48 h, and specifically, 24h, 30h, 36h, 42h or 48 h.
In the present invention, the capsule shell includes an enteric capsule shell. The enteric capsule shell can effectively resist the decomposition of gastric juice, can release effective flora in the capsule under the condition of the pH value of the intestinal tract, can effectively protect the flora from reaching the intestinal tract, prevents the flora from being released prematurely, reduces the activity loss of the flora, prolongs the release duration of the flora, achieves the specific field planting effect, and is favorable for realizing oral administration.
In the invention, the coprinus comatus capsule for children is preserved at the temperature of-75 to-80 ℃.
The invention also provides application of the anti-tumor composition in preparing a medicament for inhibiting tumor volume growth.
Specifically, said inhibiting tumor volume growth comprises inhibiting growth and/or proliferation of tumor cells.
The invention also provides application of the anti-tumor composition in preparing anti-tumor medicines.
Wherein the anti-tumor comprises inhibiting tumor volume growth and/or inhibiting migration of tumor cells.
The invention combines the children coprinus comatus capsule and the PD-1 antagonist for treating tumors, wherein the children coprinus comatus capsule and the PD-1 antagonist can be simultaneously applied or sequentially applied according to any sequence. The specific dose administered may be determined according to the specific condition of the tumor patient in combination with the clinical experience of the physician. In some embodiments of the invention, the effective dose of the coprinus comatus capsule is 2 × 106～2×108CFU/mouse, wherein the effective dose of the PD-1 antagonist is 0.09-0.1 mg/mouse. In some embodiments of the invention, the effective dose of the coprophila faecalis capsule is 2 × 108CFU/mouse, wherein the effective dose of the PD-1 antagonist is 5mg/kg of the body weight of the mouse, namely 0.09-0.1 mg/mouse. Wherein the dosage of the drug converted from the weight of 5mg/kg of mouse to each mouse is 0.09-0.1 mg/mouse calculated according to 18-20 g of the drug per mouse.
The invention also provides an anti-tumor medicament comprising the anti-tumor composition.
In the invention, the PD-1 antagonist is a PD-1 monoclonal antibody or an antigen binding fragment thereof. Wherein the PD-1 monoclonal antibody or an antigen-binding fragment thereof specifically binds to PD-1 and blocks the binding of PD-1 and PD-L1. Preferably, the PD-1 monoclonal antibody or antigen-binding fragment thereof specifically binds to human PD-1, blocking the binding of human PD-1 and human PD-L1.
The source of the PD-1 antagonist is not particularly limited in the present invention, and the antagonist can be purchased from any manufacturer. The dosage form is not particularly limited, and includes, but is not limited to, tablets, capsules, pills, granules, decoctions, pastes, lotions, oral liquids, injections, drop pills or syrups, preferably injections, such as injectable powders and injectable liquids.
The tumor in the composition, the application or the anti-tumor medicine is a solid tumor, and particularly, the tumor is a cancer expressing PD-L, including but not limited to liver cancer, colon cancer, lung cancer and melanoma. In some embodiments, the tumor is liver cancer.
Experiments show that the PD-1 antagonist and the coprinus comatus capsule are combined to treat the tumor, the curative effect is obviously higher than the effect of the two when the two are used independently, and the combination of the two produces obvious synergistic effect in the aspect of resisting the tumor.
Drawings
FIG. 1 shows the difference flora and relationship with donor before and after oral administration of fecal bacteria capsule for irritable bowel syndrome patients;
FIG. 2 is a graph of the differential PCA between healthy populations of children and adults;
FIG. 3 is a random forest modeling model of child and adult samples;
FIG. 4 is a graph showing the body weight change of mice after administration to the treatment group and the control group;
fig. 5 shows the relative tumor suppression rates of the treatment groups.
Detailed Description
The invention provides an anti-tumor composition and application thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The test materials adopted by the invention are all common commercial products and can be purchased in the market.
The invention is further illustrated by the following examples:
example 1 screening of healthy donors
extracting DNA from excrement of an excrement donor, constructing a library, and performing second-generation sequencing to obtain original sequencing data; removing host genes of original sequencing data, comparing with NCBI microbial database (bacterial genome, fungal genome, and virus genome), and performing strain identification and abundance detection; comparison with KEGG pathogen database confirmed no pathogen in the donor.
Example 2 preparation of healthy Children fecal bacteria capsules
Feces of the healthy children screened in the example 1 were collected on site, and sent to a laboratory within 1h for information registration, feces identification, weighing, evaluation and treatment, and feces bacteria liquid was prepared in an anaerobic environment, and the steps were as follows:
And 2, centrifuging the fecal filtrate at 3000r/min for 10min, and mixing the precipitate with sterile normal saline to obtain fecal bacteria liquid.
And 3, mixing the fecal strain liquid and a freeze-drying protective agent (15% of skim milk powder, 15% of trehalose, 5% of sucrose, 5% of vitamin C and the balance of normal saline) according to a ratio of 3:1(v/v), then cooling to 4 ℃ from room temperature within 10s, further cooling to-40 ℃ from 4 ℃ at a cooling speed of 2 ℃/min, cooling to-80 ℃ from-40 ℃ at a cooling speed of 5 ℃/min, cooling, freezing, then vacuum-drying for 48h under the conditions of a vacuum degree of 10pa and a vacuum degree of-50 ℃, filling the obtained fecal strain freeze-dried powder into an enteric capsule shell to obtain the fecal strain capsule, and storing at-80 ℃.
EXAMPLE 3 therapeutic Effect of fecal bacteria Capsule
Patients with irritable bowel syndrome take the fecal bacteria capsule prepared in example 2 three times a week for three weeks.
As shown in fig. 1, 16S rRNA assay was performed on a fecal sample from the patient, with samples with a significant increase in firmicutes abundance at the family classification level (p <0.01), consistent with healthy donors.
As shown in fig. 2, the results of PCA analysis of stool samples of healthy children and adults show that the stool samples of children and adults tend to have different aggregation and the flora is significantly different.
As shown in fig. 3, random forest modeling was performed on 904 samples of children and 85 samples of adults, and the results show that the model specificity is 0.91, the sensitivity is 0.89, and the modeling effect is good. Furthermore, 10 important biomarkers (biomarkers) including Bifidobacterium breve, Clostridium difficile, Bifidobacterium catenulatum, Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium kashiwanogense, Paenicilium sordelii, Bacillus steroides, Terrisporabacterium petrielareus, Blautia xlerae were selected and analyzed to be Actinomycetes and Firmicutes.
From the above results, it is clear that there is a significant difference in intestinal flora between children and adults.
Example 4 antitumor Effect of Children coprinus comatus Capsule in murine liver cancer H22 model
Experimental groups, number of animals per group and detailed routes, doses and regimens of administration are given in table 1.
Table 1 route of administration, dose and regimen in H22 animal model
Note: n: the number of animals; PD-1 is prepared into 0.5mg/ml PD-1 solution, the administration volume is 10ul/g, the administration volume of PD-1 in each mouse (the weight is calculated by 20 g) is 200ul, and the administration dosage is 0.1mg/mouse in a conversion mode, namely the administration dosage is 5 mg/kg; the dosage of GZ is 2X 108CFU/mouse。
Experimental observation indexes and calculation:
relative tumor inhibition ratio TGI (%): TGI% (1-T/C) × 100%. T/C% is the relative tumor proliferation rate, the percentage value of the relative tumor volume or tumor weight of the treated and control groups at a certain time point. T and C are the Relative Tumor Volume (RTV) or Tumor Weight (TW) at a particular time point for the treated group and the control group, respectively.
The calculation formula is as follows: T/C%RTV/CRTV×100％(TRTV: treatment group mean RTV; cRTV: control group mean RTV; RTV ═ Vt/V0，V0Is the tumor volume of the animal in the group, VtTumor volume in the animal after treatment). Or T/C%TW/CTW×100％(TTW: mean tumor weights at the end of treatment group experiments; cTW: mean tumor weights at the end of the control experiment).
H22 cells are inoculated subcutaneously in BALB/c mice to establish a subcutaneous transplantation tumor model of the mouse colon cancer. The test is divided into a test medicine of the children coprostasis capsule (2 multiplied by 10)8CFU) group, adult fecal bacteria capsule (10)8CFU), positive control anti-PD-1(5mg/kg) and solvent control, wherein each group contains 8 tested drugs, the tested drugs are orally administered for 18 days every day, and the positive control drugs are intraperitoneally administered for 8 times every week.
As shown in Table 1, the 17 th balance mean tumor volume of vehicle control mice after administration was 1148mm3. Test medicine of Children coprophila fungus capsule (2X 10)8CFU) treatment group 17 th day mean tumor volume after dosing was 1382mm3There were no statistically significant differences compared to the control group. The 17 th balance mean tumor volume of the positive control anti-PD-1(5mg/kg) treated group after administration was 476mm3Compared with the control group, the tumor inhibition ratio TGI (%) is 59 percent, and the difference is statistically significant (p is less than 0.001). The relative tumor inhibition rate of the combination group of the children fecal bacteria capsule and the anti-PD-1 is 71.4 percent, which is obviously superior to that of the single group of the children fecal bacteria capsule and the single group of the anti-PD-1. The results show that the PD-1 antagonist and the coprinus comatus capsule for children have the synergistic and enhanced effect of inhibiting the growth of tumors and have obvious curative effect on liver cancer.
TABLE 2 drug effect analysis table of each group in H22 mouse liver cancer model
As shown in fig. 3, none of the treatment groups died of animals, showed significant drug toxicity, and were well tolerated during the treatment period.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (14)
1. An anti-tumor composition comprising a PD-1 antagonist and a coprophila bassiana capsule; the children fecal bacteria capsule is prepared from feces of healthy children of 7-16 years old.
2. The anti-tumor composition according to claim 1, wherein the ratio of the mass of the PD-1 antagonist to the colony count of the fecal bacteria capsule for children is (0.09-0.1) mg (2 x 10)6～2×108)CFU。
3. The anti-tumor composition according to claim 1, wherein the ratio of the mass of the PD-1 antagonist to the colony count of the microcapsule is 0.1:2 x 10 in mg/CFU8。
4. The anti-tumor composition according to claim 1, wherein the coprophila bassiana capsule is prepared by the following method:
step 1, selecting healthy children of 7-16 years old, and carrying out health questionnaire investigation to obtain qualified donors after primary screening;
2, screening the qualified donors by utilizing a high-throughput sequencing method;
step 3, carrying out clinical examination on the donor screened in the step 2 to obtain a target donor;
step 4, soaking the feces of the target donor screened in the step 3 in sterile physiological saline, filtering, centrifuging, and mixing the precipitate with the sterile physiological saline to obtain a fecal bacteria liquid;
and 5, mixing the fecal bacteria liquid prepared in the step 4 with a freeze-drying protective agent, cooling, freezing and vacuum-drying to obtain fecal bacteria freeze-dried powder, and filling the obtained fecal bacteria freeze-dried powder into a capsule shell to obtain the children fecal bacteria capsule.
5. The anti-tumor composition according to claim 4, wherein the method for screening donors by high throughput sequencing in step 2 comprises the following steps:
(1) performing second-generation sequencing on the DNA and/or RNA extracted from the donor to obtain original sequencing data;
(2) removing host genes of the original sequencing data, comparing the host genes with a microbial database, and performing strain identification and abundance detection;
(3) comparing with a pathogen database to confirm that no pathogen exists in the donor;
2, screening the fecal bacteria donor by using high-throughput sequencing according to the biomarkers expressed by the fecal bacteria donor and the alpha diversity index of the biomarkers;
the biomarker includes at least one of escherichia coli, clostridium polybotrys, eubacterium columnare, human rhoebergia, clostridium praecox, bacteroides fragilis or bacteroides vulgatus.
6. Use of the anti-tumor composition of any one of claims 1 to 5 for the preparation of a medicament for inhibiting tumor volume growth.
7. The use of claim 6, wherein the inhibiting tumor volume growth comprises inhibiting growth and/or proliferation of tumor cells.
8. Use of the composition of any one of claims 1 to 5 in the preparation of an anti-tumor medicament.
9. The use of claim 6, wherein the anti-tumor comprises inhibiting tumor volume growth and/or inhibiting tumor cell migration.
10. The use according to any one of claims 4 to 9, wherein the effective dose of the coprophila bassiana capsule is 2 x 106～2×108CFU/mouse, wherein the effective dose of the PD-1 antagonist is 0.09-0.1 mg/mouse.
11. An antitumor agent comprising the antitumor composition according to any one of claims 1 to 5.
12. The antitumor drug as claimed in claim 11, wherein the PD-1 antagonist and pharmaceutically acceptable excipients are formulated into a dosage form selected from the group consisting of tablets, capsules, pills, granules, decoctions, pastes, lotions, oral liquids, injections, drop pills and syrups.
13. The composition of any one of claims 1 to 5 or the use of any one of claims 6 to 10 or the anti-neoplastic agent of any one of claims 11 to 12, wherein the PD-1 antagonist is a PD-1 monoclonal antibody or an antigen-binding fragment thereof.
14. The composition according to any one of claims 1 to 5, the use according to any one of claims 6 to 10, or the antitumor agent according to any one of claims 11 to 12, wherein the tumor is liver cancer, intestinal cancer, lung cancer, or melanoma.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110582291A (en) * | 2016-12-22 | 2019-12-17 | 古斯达威罗斯研究所 | Reactivity marker and efficacy-improving modulator for anti-PD 1/PD-L1/PD-L2 antibody |
| CN111249314A (en) * | 2020-03-03 | 2020-06-09 | 深圳未知君生物科技有限公司 | Role of human symbiotic flora in improving tumor immunotherapy response |
| CN112089736A (en) * | 2020-09-22 | 2020-12-18 | 苏州普瑞森基因科技有限公司 | Whole fungus capsule and preparation method and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105193858A (en) * | 2015-09-25 | 2015-12-30 | 南昌大学 | Application of natural faeces flora transplanting in intestinal disease treatment |
| US20210361721A1 (en) * | 2018-03-28 | 2021-11-25 | Seres Therapeutics, Inc. | Treatment of a cancer by microbiome modulation |
| WO2020205927A2 (en) * | 2019-04-01 | 2020-10-08 | Board Of Regents, The University Of Texas System | Tumor microbiome signature and therapeutic use of fecal microbiota transplantation on pancreatic cancer patients |
| CN114432439A (en) * | 2021-12-07 | 2022-05-06 | 苏州普瑞森生物科技有限公司 | Anti-tumor composition and application thereof |
-
2021
- 2021-12-07 CN CN202111487574.2A patent/CN114432439A/en active Pending
-
2022
- 2022-08-17 WO PCT/CN2022/113005 patent/WO2023103441A1/en unknown
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110582291A (en) * | 2016-12-22 | 2019-12-17 | 古斯达威罗斯研究所 | Reactivity marker and efficacy-improving modulator for anti-PD 1/PD-L1/PD-L2 antibody |
| US20210346438A1 (en) * | 2016-12-22 | 2021-11-11 | Institut Gustave Roussy | Microbiota composition, as a marker of responsiveness to anti-pd1/pd-l1/pd-l2 antibodies and use of microbial modulators for improving the efficacy of an anti-pd1/pd-l1/pd-l2 ab-based treatment |
| CN111249314A (en) * | 2020-03-03 | 2020-06-09 | 深圳未知君生物科技有限公司 | Role of human symbiotic flora in improving tumor immunotherapy response |
| CN112089736A (en) * | 2020-09-22 | 2020-12-18 | 苏州普瑞森基因科技有限公司 | Whole fungus capsule and preparation method and application thereof |
Non-Patent Citations (7)
| Title |
|---|
| GAOCHEN LU等: "Diversified aspects of donor screening for washed microbiota transplantation at china microbiota transplantation system", GUT, vol. 70, no. 2, pages 112 * |
| 吴震: "脊索瘤", 31 August 2020, 科学技术出版社, pages: 56 - 57 * |
| 张毅: "肿瘤生物治疗临床应用", 30 April 2020, 河南科学技术出版社, pages: 107 * |
| 杨梦雪;严雪冰;王颖;童建东;: "肠道菌群在肿瘤免疫治疗中的作用研究进展", 生命的化学, no. 08 * |
| 池肇春等: "肠道微生物与消化***疾病", 30 June 2020, 上海科学技术出版社, pages: 338 - 341 * |
| 谷野等: "肝癌的多学科综合诊断与治疗", 30 September 2020, 辽宁科学技术出版社, pages: 156 * |
| 赵家军等: "***内分泌学", 31 July 2020, 科学普及出版社, pages: 1505 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023103441A1 (en) * | 2021-12-07 | 2023-06-15 | 苏州普瑞森生物科技有限公司 | Anti-tumor composition and use thereof |
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| WO2023103441A1 (en) | 2023-06-15 |
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