Composition containing oncolytic virus and application of composition in preparation of medicine with anti-tumor effect

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a composition containing oncolytic viruses and application of the composition in preparation of a medicine with an anti-tumor effect. The composition containing the oncolytic virus comprises a phloroglucinol compound and the oncolytic virus. The phloroglucinol compound is Gammultinone A, and the structural formula of the phloroglucinol compound is shown in the description. The oncolytic virus is selected from a VSV [delta] 51 oncolytic virus. The research shows that after the phloroglucinol compound Garmultinone A is combined with the oncolytic virus, the antitumor activity of the oncolytic virus can be greatly improved; the antitumor activity of the compound is greatly higher than that of an oncolytic virus independently used, and is also greatly higher than that of a Garmultinone A independently used.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a composition containing an oncolytic virus and an application thereof in the preparation of a drug with an anti-tumor effect. Background Art

[0002] Oncolytic viruses are a type of replicable virus that selectively infects and kills tumor cells without damaging normal cells. Oncolytic virotherapy is an innovative tumor-targeted treatment strategy that uses natural or genetically engineered viruses to selectively infect tumor cells and replicate in tumor cells, achieving targeted oncolysis and tumor cell killing without damaging normal cells.

[0003] Vesicular stomatitis virus (VSV) belongs to the Rhabdoviridae family and is a negative-strand RNA virus. VSVΔ51 is a recombinant oncolytic virus that lacks the 51st methionine residue of the M protein of the VSV virus. This amino acid deletion can block the impaired interferon response caused by viral infection in normal cells and increase the safety of VSV in normal tissues. VSVΔ51 has a good application effect in the preparation of anti-tumor drugs. However, different tumors have different sensitivities to the VSVΔ51 oncolytic virus. For some tumors, the oncolytic effect of the VSVΔ51 oncolytic virus is not ideal when used alone.

[0004] Therefore, how to further improve the anti-tumor effect of VSVΔ51 oncolytic virus is a technical problem that technicians in this field need to further solve.

[0005] Garmultinone A is a known compound, which has been disclosed in the document "Chen Y, Ma Z, Teng H, et al. Fourintricately caged polycyclic polyprenylated acylphloroglucinols from Garciniamultiflora fruits [J]. Organic Chemistry Frontiers, 2019, 6 (17): 3085-3092."; however, the prior art does not disclose that Garmultinone A has the ability to enhance the anti-tumor effect of VSVΔ51 oncolytic virus. Summary of the invention

[0006] In order to overcome at least one technical problem existing in the prior art, the present invention provides an application of a composition containing an oncolytic virus in the preparation of a drug with anti-tumor effect.

[0007] The technical solution of the present invention is as follows:

[0008] The present invention first provides an oncolytic virus-containing composition, which comprises a phloroglucinol compound and an oncolytic virus.

[0011] Preferably, the phloroglucinol compound is Garmultinone A.

[0012] The present invention provides a novel composition containing oncolytic viruses. The inventors have found in experimental screening that not all compounds can enhance the anti-tumor activity of oncolytic viruses. The inventors have surprisingly found in a large number of experiments that the combination of phloroglucinol compound Garmultinone A and oncolytic viruses can significantly enhance the anti-tumor activity of oncolytic viruses. The anti-tumor activity is significantly higher than that of oncolytic viruses alone, and is also significantly higher than that of Garmultinone A alone.

[0013] Preferably, the dosage ratio of the mesoglucosidol compound to the oncolytic virus in the composition is 25-100 mg: 1×10 7 -10 8 PFU.

[0014] Most preferably, the ratio of the amount of the mesotriol compound to the oncolytic virus in the composition is 75 mg:1×10 7 PFU.

[0015] Preferably, the oncolytic virus is selected from VSVΔ51 oncolytic virus.

[0016] Further preferably, the VSVΔ51 oncolytic virus is selected from the Indian strain, and its matrix protein (M)

[0017] No.

[0018] The amino acid methionine at position 51 was knocked out.

[0019] The present invention also provides a use of the oncolytic virus composition in the preparation of a drug with anti-tumor effect.

[0020] Preferably, the tumor is a solid tumor.

[0021] More preferably, the solid tumor is glioma, breast cancer, liver cancer, colorectal cancer, bladder cancer,

[0022] cervix

[0023] cancer, prostate cancer, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, or stomach cancer.

[0024] Preferably, the drug is in the form of an injection, tablet, capsule, patch or kit.

[0025] Preferably, the dosage form of the drug is intravenous injection.

[0026] Beneficial effects: The present invention provides a new composition; the combination of the phloroglucinol compound Garmultinone A and oncolytic virus can greatly improve the anti-tumor activity of the oncolytic virus; its anti-tumor activity is much higher than that of the oncolytic virus alone, and is also much higher than that of Garmultinone A alone.

[0027] The specific experimental research conclusions of the present invention are as follows:

[0028] Garmultinone A itself has extremely low toxicity to cells, and its combined use with oncolytic viruses can significantly increase the anti-tumor effect of oncolytic viruses, thereby improving the therapeutic effectiveness of oncolytic viruses as anti-tumor drugs. Cytological experiments have shown that the combined use of VSVΔ51 oncolytic virus and Garmultinone A can significantly cause morphological changes in tumor cells and induce apoptosis of tumor cells, thereby significantly enhancing the inhibitory effect on tumor cells.

[0029] We combined Garmultinone A and VSVΔ51 oncolytic virus to act on human glioma cells LN229, and found that when the small molecule compound Garmultinone A and VSVΔ51 oncolytic virus were used in combination, the morphological changes of tumor cells were significantly increased and the survival rate of tumor cells was significantly reduced. For example, in one embodiment of the present invention, when Garmultinone A (8μM) and VSVΔ51 (MOI = 0.001) were used alone to treat glioma cells, the survival rates of tumor cells were 96.1% and 82.8%, respectively, and when 8μM Garmultinone A was used in combination with 0.001MOI VSVΔ51 virus, the survival rate of tumor cells dropped significantly to 60.1%. Compared with the anti-tumor effect of VSVΔ51 virus alone, the oncolytic effect was significantly improved when Garmultinone A was used in combination with VSVΔ51.

[0030] In addition, we found that among the more than 80 compounds we screened, not all of them could enhance the anti-tumor effect of VSVΔ51 oncolytic virus. Only Garmultinone A could significantly enhance the anti-tumor effect of VSVΔ51 oncolytic virus. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1This is a graph showing the experimental results showing that Garmultinone A and VSVΔ51 viruses significantly increased the morphological lesions of human glioma cell lines.

[0032] Figure 2 This is the experimental result of the combined treatment of Garmultinone A and VSVΔ51 virus significantly reducing the survival rate of tumor cell cancer lines, among which, Figure 2 a The combined treatment of Garmultinone A and VSVΔ51 virus significantly reduced the survival rate of human glioma cell line LN229. Figure 2 b Combined treatment of Garmultinone A and VSVΔ51 virus significantly reduced the survival rate of human breast cancer cell line MDA-MB-231.

[0033] Figure 3 This is the experimental result of promoting apoptosis of human glioma cell line LN229 and human breast cancer cell line MDA-MB-231 by combined treatment of Garmultinone A and VSVΔ51 virus. Figure 3 a, Annexin V / PI apoptosis kit to detect cell apoptosis, Figure 3 b, Detection of the protein level of apoptosis protein Cleaved-caspase3.

[0034] Figure 4 This is the experimental result showing that the combined treatment of Garmultinone A and VSVΔ51 virus significantly inhibited the growth of transplanted tumors in glioma cell lines. Figure 4 a, Garmultinone A combined with VSVΔ51 virus treatment significantly inhibited the growth of glioma cell line transplanted tumors; Figure 4 b, There was no significant change in the body weight of mice treated with Garmultinone A and VSVΔ51 virus, proving the safety of this treatment method; Figure 4 c, Immunohistochemistry analysis (IHC) demonstrated that the combined treatment of Garmultinone A and VSVΔ51 virus significantly inhibited the growth of glioma cell line transplanted tumors.

[0035] Figures 1 to 4 Explanation of the icons: GA: Garmultinone A treatment group; VSVΔ51: VSVΔ51 oncolytic virus treatment group; VSVΔ51+GA: VSVΔ51 and Garmultinone A combined treatment group. DETAILED DESCRIPTION

[0036] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

[0037] Example 1 Preparation of Garmultinone A

[0038] (1) 30 kg of dried fruit peel of Lingnan Mountain Bamboo was crushed into coarse powder, passed through an 80-mesh sieve, and extracted by dichloromethane percolation. The extraction was repeated three times for 24 hours each time. The extracts were combined and concentrated under reduced pressure to obtain 2.3 kg of dichloromethane total extract.

[0039] (2) After the total dichloromethane extract was dissolved in dichloromethane, it was mixed with 2.3 kg of silica gel (80-100 mesh) and evaporated to dryness under reduced pressure to obtain a sample. Another 15 kg of silica gel (200-300 mesh) was wet-packed into a column (inner diameter × length = 10 × 75 cm), the sample was loaded onto the column, and then covered with a layer of silica gel, first eluted with dichloromethane, and then gradient eluted with dichloromethane and methanol (100:0→0:100); according to the TLC analysis results, the eluate was heated and concentrated under reduced pressure on a rotary evaporator to obtain 6 fractions.

[0040] (3) The first fraction was dissolved in methanol, mixed with 200 g of silica gel (80-100 mesh), and evaporated to dryness under reduced pressure to obtain a sample. Another 2 kg of silica gel (200-300 mesh) was wet-packed into a column (inner diameter × length = 10 × 50 cm), the sample was loaded onto the column, and then covered with a layer of silica gel, first eluted with petroleum ether, and then gradient eluted with petroleum ether and ethyl acetate (100:0→0:100); according to the TLC analysis results, the eluate was heated and concentrated under reduced pressure on a rotary evaporator to obtain 5 fractions.

[0041] (4) The third fraction was dissolved in methanol, mixed with 50 g of silica gel (80-100 mesh), and evaporated to dryness under reduced pressure to obtain a sample. Another 1.5 kg of silica gel (200-300 mesh) was wet-packed into a column (inner diameter × length = 15 × 35 cm), the sample was loaded onto the column, and then covered with a layer of silica gel, and gradient eluted with methanol and water (30:70 → 100:0) to obtain a mixed solution of phloroglucinol compounds.

[0042] (5) The mixed solution was dissolved in methanol and loaded onto a Sephadex LH-20 gel column with a column volume of 50 ml. The column was followed and analyzed by an analytical high performance liquid chromatography (HPLC). The column was eluted isocratically with methanol: water (90:10). The fractions with a retention time of 15-30 min were collected and concentrated under reduced pressure to obtain 1.4 g of a crude extract of phloroglucinol compounds.

[0043] (6) The crude extract of phloroglucinol compounds was prepared by preparative HPLC to obtain Garmultinone A; the specific conditions of the preparative HPLC were: 90% methanol water, collecting the fraction corresponding to the chromatographic peak at 5.6 min, concentrating and drying to obtain the Garmultinone A.

[0044] Analysis of Garmultinone A: White powder. The color reaction is green (vanillin-ethanol-concentrated sulfuric acid). The UV spectrum (CH3OH) shows maximum absorption peaks at 203nm, 263nm, and 325nm. IR (KBr) shows that the structure contains hydroxyl groups (3436cm -1 ), double bond (2971cm -1 ,2912cm -1 ), carbonyl (1729cm -1 ), benzene ring (1613cm -1 ). HR-ESI-MS m / z541.2938[M+Na] + (ca lcd for C 33 H 42 O5Na,541.2924), further analysis of its 13 C NMR signal, the molecular formula is inferred to be C 33 H 42 O5,Ω=13. 1 H NMR spectrum (400MHz, CDCl3) sorted out the hydrogen spectrum signals of compound Garmultinone A: [δ H 1.63 (3H, s), 1.57 (3H, s), 1.56 (3H, s), 1.28 (3H, s), 1.06 (3H, s), 0.95 (3H, s), 0.94 (3H, s)] are 7 methyl proton signals, [δ H 4.99 (1H, t, J = 6.4 Hz), 4.93 (1H, t, J = 6.2 Hz)] are two olefin proton signals, [δ H 7.51 (1H, d, J = 7.6 Hz), 7.42 (1H, d, J = 7.2 Hz), 7.31 (1H, t, J = 7.3 Hz)] are 5 benzene ring proton signals. Comprehensive analysis of the DEPT-135 spectrum shows that the 33 carbon signals of Garmultinone A include 7 primary carbons, 5 secondary carbons, 9 tertiary carbons and 12 quaternary carbons. Two carbonyl groups, one benzene ring and three pairs of double bonds were identified based on the chemical shift characteristics. After retrieval and signal comparison, it was determined to be the known compound Garmultinone A; the chemical structure of Garmultinone A is shown in Formula I:

[0045]

[0046] The mass spectrum and NMR data of Garmultinone A are as follows: HR-ESI-MS m / z 541.2938[M+Na] + (calcdfor C 33 H 42O5Na,541.2924)

[0047] Table 1NMR data of Garmultinone A(400MHz)a

[0048]

[0049]

[0050] Example 2 Garmultinone A and VSVΔ51 virus significantly increased the morphological changes of human glioma cell lines

[0051] Materials: Human glioma cells LN229, high-glucose DMEM medium, inverted phase contrast microscope.

[0052] method:

[0053] a) Cell culture: Human glioma cells LN229 were grown in DMEM complete medium containing 10% FBS, 100U / ml penicillin and 0.1mg / ml streptomycin; all cell lines were cultured and passaged in a 5% CO2, 37°C constant temperature sealed incubator (relative humidity 95%), and the growth was observed under an inverted microscope. The cells were passaged approximately every 2-3 days, and cells in the logarithmic growth phase were used for formal experiments.

[0054] b) Cell treatment and morphological observation: Cells in logarithmic growth phase were selected and cell suspension was prepared with DMEM complete culture medium (containing 10% fetal bovine serum and 1% double antibody). 4 The cells were inoculated at a density of 100 μg / well in a 24-well culture plate. Cells were treated with Garmultinone A (8 μM) alone, infected with VSVΔ51 virus (MOI=0.001), and treated with virus (MOI=0.001) combined with Garmultinone A (8 μM). Cells without VSVΔ51 virus and Garmultinone A were used as controls. After 48 hours, the changes in cell morphology were observed under an inverted phase contrast microscope.

[0055] result:

[0056] like Figure 1As shown, the cell morphology was observed under a phase contrast microscope. LN229 cells grew in a monolayer and were closely arranged. The cell morphology of the control group was consistent with that of Garmultinone A (8μM), and there was no significant change. However, after 48 hours of treatment with the VSVΔ51 virus (MOI = 0.001), the cell morphology changed significantly. Compared with the cells in the control group and the VSVΔ51 alone treatment group, the number of cells in the combined treatment group was significantly reduced, the cell bodies shrank into a spherical shape, the refractive index was significantly enhanced, and it was a dead lesion. This also shows that the phloroglucinol compound Garmultinone A can significantly improve the anti-tumor activity of the oncolytic virus after combined use with the oncolytic virus; its anti-tumor activity is much higher than that of the oncolytic virus alone, and also much higher than that of Garmultinone A alone.

[0057] Example 3 Garmultinone A combined with VSVΔ51 virus treatment significantly reduces the survival rate of human glioma cell lines and human breast cancer cell lines

[0058] Materials: Human glioma cells LN229, human breast cancer cells MDA-MB-231, VSVΔ51 virus, high-glucose DMEM medium, automatic enzyme-linked detection microplate reader

[0059] method:

[0060] a) Cell inoculation and drug treatment: cells in logarithmic growth phase were selected and cell suspension was prepared with DMEM complete culture medium (containing 10% fetal bovine serum and 1% double antibody). 3 / well were inoculated in a 96-well culture plate. After 12 hours, the cells were completely attached to the wall. The experiment was divided into a control group without treatment, a Garmultinone A alone group, a VSVΔ51 alone infection group, and a Garmultinone A and VSVΔ51 combined group. The doses used were: VSVΔ51 virus (MOI = 0.001) infected cells; Garmultinone A set different dose gradients.

[0061] b) MTT reacts with succinate dehydrogenase in cells: After 48 hours of culture, add 20 μl (5 mg / ml) of MTT to each well and continue incubation for 4 hours. At this time, granular blue-purple formazan crystals can be observed in living cells under a microscope.

[0062] c) Dissolve the formazan particles: Carefully remove the supernatant, add DMSO 100μl / well to dissolve the formed crystals, shake on a micro-oscillator for 5 minutes, and then measure the optical density (OD value) of each well on an enzyme-linked detector at a wavelength of 570nm. Repeat each group of experiments 3 times. Cell survival rate = OD value of drug-treated group / OD value of control group × 100%.

[0063] d) Graphpad software was used for nonlinear curve fitting. Two dose-effect curves were drawn with drug dose as the horizontal axis and relative cell survival rate as the vertical axis, namely the dose-effect curve of Garmultinone A alone and the dose-effect curve of Garmultinone A combined with VSVΔ51 virus. The difference in the area under the curve of the two curves, namely DAUC (Differnence in Area Under the Curve, DAUC) in the figure, was calculated. The larger the difference, the more significant the drug combination effect. The survival rate of cells after various treatments was divided by the survival rate of the control group without drug and virus treatment as the relative cell survival rate, and expressed as a multiple. One-way ANOVA statistics indicated that there was a statistical difference. * indicates P < 0.05 and ** indicates P < 0.01.

[0064] result:

[0065] like Figure 2 As shown in a, treatment with VSVΔ51 virus (MOI=0.001) alone had a small inhibitory effect on the survival rate of tumor cells LN229, and the relative cell survival rate of tumor cells was 82.8%. The relative cell survival rate of tumor cells in the group treated with 8μM Garmultinone A was still as high as 96.1%. However, when the same 8μM Garmultinone A was used in combination with VSVΔ51 virus (MOI=0.001) (Garmultinone A+VSVΔ51), the relative cell survival rate of tumor cells dropped significantly to 60.1%. Figure 2 As shown in b, the relative cell survival rate of tumor cells MDA-MB-231 treated with VSVΔ51 virus (MOI=0.001) alone was 72.3%, and the relative cell survival rate of tumor cells in the group treated with 8μM Garmultinone A was still as high as 96.9%. However, when the same 8μM Garmultinone A was used in combination with VSVΔ51 virus (MOI=0.001) (Garmultinone A+VSVΔ51), the relative cell survival rate of tumor cells dropped significantly to 50.9%. Compared with the single treatment, the combined use of GarmultinoneA and VSVΔ51 virus (MOI=0.001) significantly reduced the survival rate of tumor cells LN229 and MDA-MB-231, indicating that Garmultinone A can enhance the oncolytic effect of VSVΔ51 virus at the cellular level. This also shows that the combination of the phloroglucinol compound Garmultinone A and oncolytic virus can greatly enhance the anti-tumor activity of oncolytic virus; its anti-tumor activity is much higher than that of oncolytic virus alone, and also much higher than that of Garmultinone A alone.

[0066] Example 4: Combined treatment of Garmultinone A and VSVΔ51 virus induces apoptosis of tumor cells

[0067] Materials: Garmultinone A, VSVΔ51 virus, human glioma cells LN229, human breast cancer cells MDA-MB-231, Annexin V / PI apoptosis kit (Biolegend, USA), trypsin, PBS, Western blot: cell IP lysis buffer (Beyotime, China), α-Tubulin antibody (Proteintech, China), Caspase3 antibody (Proteintech, China), Cleaved-capase3 antibody (Proteintech, China)

[0068] method:

[0069] a) Annexin V / PI apoptosis flow cytometry: cells in logarithmic growth phase were selected and cell suspension was prepared with high-glucose DMEM medium (containing 10% fetal bovine serum and 1% double antibody). 5 / well were inoculated in a 6-well plate. After 24 hours, the cells were completely attached to the wall. The experiment was divided into blank tubes, single-stained tubes, negative tubes and experimental groups (control group, Garmultinone A group, VSVΔ51 group and Garmultinone A / VSVΔ51 combination group), among which VSVΔ51 did not carry GFP. The dosage used was: VSVΔ51 virus (MOI = 0.001) infected cells; Garmultinone A (8 μM) treated cells. After 48 hours, the cell supernatants were collected, the cells were digested with EDTA-free trypsin, and collected in centrifuge tubes, centrifuged at 1000 rpm for 3 minutes, the supernatant was discarded, and the cells were washed twice with pre-cooled PBS, each time at 4°C 1000 rpm for 3 minutes, the supernatant was discarded, and Annexin V Binding Buffer was added to adjust the cells to the same concentration (generally 1×10 6 / mL), add 2.5ul FITC Annexin V and 5ul Propidiumlodide solution respectively, vortex the cells gently, incubate at room temperature in the dark for 15 minutes, add 300ul Annexin V BindingBuffer to each tube, and detect on the instrument.

[0070] b) Protein samples were extracted and the expression of Caspase 3 and Cleaved-capase3 proteins was detected by Western blot, with α-Tubulin as the internal reference.

[0071] c) The experiment was repeated three times, and the data are expressed as mean ± SD. The data were compared with the respective control groups using the student's t test, and *** indicates P < 0.001.

[0072] result:

[0073] like Figure 3 As shown in Figures 3a and 3b, in human glioma LN229 cells and human breast cancer MDA-MB-231 cells, the rate of cell apoptosis in the Garmultinone A / VSVΔ51 combination group was significantly increased compared with the Garmultinone A group and the VSVΔ51 group alone. Figure 3 As shown in c, the bottom of the Western blot image shows that in human glioma cells LN229 and human breast cancer MDA-MB-231 cells, the expression of Cleaved-capase 3 protein in the Garmultinone A / VSVΔ51 combination group was significantly higher than that in the Garmultinone A group and the VSVΔ51 group. This also shows that the combination of phloroglucinol compound Garmultinone A and oncolytic virus can greatly improve the anti-tumor activity of oncolytic virus; its anti-tumor activity is much higher than that of oncolytic virus alone, and also much higher than that of Garmultinone A alone.

[0074] Example 5 The combined use of Garmultinone A and VSVΔ51 virus significantly inhibits the growth of transplanted tumors of glioma cell lines

[0075] Materials: Garmultinone A, VSVΔ51 virus, glioma cell line GL261, and 4-week-old female BALB / c nude mice.

[0076] method:

[0077] a) This experiment adopts a randomized, single-blind design. 7 GL261 cells were injected subcutaneously into the dorsal flank of 4-week-old BALB / c nude mice. When the tumor size reached 100 mm 3 The rats were divided into groups according to the time of treatment, including the control group without treatment, the group with Garmultinone A alone (intraperitoneal injection of 75 mg / kg / day), the group with VSVΔ51 alone (injection of VSVΔ51 virus 1×10 7 PFU / time) and Garmultinone A / VSVΔ51 combination group (the same doses of Garmultinone A and VSVΔ51 were given in the same manner, Garmultinone A was injected continuously for 7 times, and VSVΔ51 was injected continuously for the last 3 days).

[0078] b) The length, width and body weight of the tumor were measured every two days, and the tumor volume was calculated according to the formula (length × width × 2) / 2. One-way ANOVA statistics were performed after measuring the tumor volume, *** indicates P < 0.001, ** indicates P < 0.01, and * indicates P < 0.05.

[0079] c) Immunohistochemical analysis (IHC): On the second day after the end of drug administration, mice were anesthetized and killed, tumor tissues were separated, paraffin sections were made, and the sections were baked at 60°C for 30 minutes, dewaxed and hydrated; 3% methanol-H2O2 was used for 20 minutes to inactivate endogenous peroxidase; the sections were immersed in boiling 0.01M citrate buffer, treated at low heat for 6 minutes, stopped for 6 minutes, repeated four times, and then naturally cooled to room temperature, antigen repair, and blocked with normal non-immune animal serum for 20 minutes; the blocking solution was discarded, and primary antibodies (Ki67, Cleaved-capase3) were added to completely cover the specimens, and kept at 4°C overnight; the sections were rewarmed at 37°C for 45 minutes, and the DAB ready-to-use detection system was used for color development, hematoxylin counterstaining, water washing, and differentiation followed by thorough water washing; dehydration, transparency, neutral glue sealing, and drying at 50°C for 1-2 hours.

[0080] result:

[0081] In tumor cell transplanted tumor animals, tumor volume was measured by vernier caliper. The results showed that compared with the control group, the group infected with Garmultinone A and VSVΔ51 alone could only cause a slight reduction in tumor volume, while the group infected with GarmultinoneA / VSVΔ51 combined could cause a significant reduction in tumor volume ( Figure 4 a), and One way ANOVA statistics showed that there was a statistical difference ( Figure 4 a), this synergistic effect was particularly amplified in tumors that were less sensitive to oncolytic virus alone, with a dramatic reduction in tumor volume ( Figure 4 a), and the body weight of mice did not change significantly ( Figure 4 b). IHC analysis showed that the VSVΔ51 infection group alone could only cause a slight decrease in Ki67 and a slight increase in Cleaved-capase3 expression, while the Garmultinone A / VSVΔ51 combination group could cause a significant decrease in Ki67 and a significant increase in Cleaved-capase3 ( Figure 4 c). This also shows that: the combination of phloroglucinol compound Garmultinone A and oncolytic virus can significantly improve the anti-tumor activity of oncolytic virus; its anti-tumor activity is significantly higher than that of oncolytic virus alone, and also significantly higher than that of Garmultinone A alone.

Claims

1. A composition containing an oncolytic virus, characterized in that: Contains phloroglucinol compounds and oncolytic viruses.

2. The composition containing oncolytic virus according to claim 1, characterized in that The phloroglucinol compound is GarmultinoneA.

3. The composition containing oncolytic virus according to claim 1, characterized in that The dosage ratio of the mesotriol compound to the oncolytic virus in the composition is 25-100 mg: 1×10 7 -10 8 PFU; Most preferably, the ratio of the amount of the mesotriol compound to the oncolytic virus in the composition is 75 mg:1×10 7 PFU.

4. The composition containing oncolytic virus according to claim 1, characterized in that The oncolytic virus is selected from VSVΔ51 oncolytic virus.

5. The oncolytic virus composition according to claim 1, characterized in that The VSVΔ51 oncolytic virus is selected from the Indian strain, and the 51st amino acid methionine of its matrix protein (M) is knocked out.

6. Use of the oncolytic virus composition according to any one of claims 1 to 5 in the preparation of a drug with anti-tumor effect.

7. The use according to claim 6, characterized in that: The tumor is a solid tumor.

8. The use according to claim 7, characterized in that: The solid tumor is glioma, breast cancer, liver cancer, colorectal cancer, bladder cancer, cervical cancer, prostate cancer, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer or gastric cancer.

9. The use according to claim 7, characterized in that: The dosage form of the medicine is injection, tablet, capsule, patch or kit.

10. The use according to claim 9, characterized in that: The dosage form of the drug is intravenous injection.

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