USE OF PROTEASOME INHIBITOR AND ALPHAVIRUS IN PREPARATION OF ANTI-TUMOR MEDICAMENT

Abstract

Use of a proteasome inhibitor and an alphavirus in the preparation of an anti-tumor medicament. The proteasome inhibitor can be used to prepare an alphavirus anti-tumor synergist. A pharmaceutical composition comprising a proteasome inhibitor and an alphavirus, including a pharmaceutical kit comprising the proteasome inhibitor and the alphavirus, and use of the proteasome inhibitor and the virus in the treatment of tumors, particularly tumors insensitive to the alphavirus.

Claims

1-10. (canceled)

11. A method for treating a tumor in a subject in need thereof, comprising: administering to the subject in need thereof an effective amount of a proteasome inhibitor, and an effective amount of an alphavirus.

12. The method of claim 11, wherein the alphavirus is selected from the group consisting of Eastern Equine Encephalitis virus, Venezuelan Equine Encephalitis virus, Everglades virus, Mucambo virus, Pixuna virus, Western Encephalitis virus, Sindbis virus, South African arbovirus No. 86, Girdwood S. A. virus, Ockelbo virus, Semliki Forest virus, Middleburg virus, Chikungunya virus, O'Nyong-Nyong virus, Ross River virus, Barmah Forest virus, Sagiyama virus, Bebaru virus, Mayaro virus, Una virus, Aura virus, Whataroa virus, Babanki virus, Kyzlagach virus, Highlands J virus, Fort Morgan virus, Ndumu virus, Buggy Creek virus, M1 virus and Getah virus.

13. The method of claim 11, wherein the alphavirus is selected from at least one of M1 virus and Getah virus.

14. The method of claim 11, wherein the alphavirus is M1 virus.

15. The method of claim 11, wherein the genome sequence of the alphavirus has at least 95% identity to the sequence indicated by Genbank Accession No. EF011023; and/or the genome sequence of the alphavirus has at least 95% identity to the genome sequence of the virus deposited under accession No. CCTCC V201423.

16. The method of claim 11, wherein the proteasome inhibitor is a substance that inhibits proteasome activity, or inhibits the activity or expression of any one subunit of the proteasome, or blocks assembly of proteasome subunits, or degrades the proteasome.

17. The method of claim 11, wherein the proteasome inhibitor is selected from a group consisting of: Bortezomib, Carfilzomib, MG-132, ONX-0914, ONX-0912, CEP-18770, MLN-9708, Epoxomicin, VR23, MLN-2238, Celastrol and P1-18400; or derivatives thereof having proteasome inhibitory effect, or pharmaceutically acceptable salts, solvates, tautomers, isomers thereof.

18. The method of claim 11, wherein the proteasome inhibitor is selected from a group consisting of: Bortezomib, Carfilzomib, MG-132, ONX-0914, ONX-0912, CEP-18770 and MLN-9708; or derivatives thereof having proteasome inhibitory effect, or pharmaceutically acceptable salts, solvates, tautomers, isomers thereof.

19. The method of claim 11, wherein the proteasome inhibitor is selected from gene interference, gene editing, gene silencing or gene knockout materials.

20. The method of claim 11, wherein the proteasome inhibitor is selected from one or more of DNA, RNA, PNA and DNA-RNA hybrids.

21. The method of claim 11, wherein the proteasome inhibitor is selected from one or more of siRNA, dsRNA, miRNA, shRNA and ribozyme.

22. The method of claim 11, wherein the proteasome inhibitor is a tumor targeting proteasome inhibitor.

23. The method of claim 11, wherein the tumor is a solid tumor or a hematological tumor.

24. The method of claim 11, wherein the tumor is selected from a group consisting of: adrenocortical carcinoma, pararenocortical carcinoma, anal carcinoma, appendiceal carcinoma, astrocytoma, atypical teratoma, rhabdomyoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain tumor, bronchial tumor, Burkett's lymphoma, carcinoid tumor, heart tumor, bile duct epithelial carcinoma, chordoma, colorectal cancer, craniopharyngioma, ductal carcinoma in situ, embryonal tumor, endometrial carcinoma, ependymoma, esophageal carcinoma, olfactory neuroblastoma, intracranial embryonic cell tumor, extragonadal germ cell tumor, eye cancer, carcinoma of the fallopian tube, gallbladder carcinoma, head and neck cancer, hypopharyngeal carcinoma, Kaposi's sarcoma, renal carcinoma, Langerhans cell histiocytosis, laryngeal carcinoma, lip cancer, oral cancer, Meckel cell carcinoma, malignant mesothelioma, multiple endocrine neoplasia syndrome, mycosis fungoides, nasal sinus carcinoma, neuroblastoma, non-small cell lung cancer, ovarian cancer, pancreatic neuroendocrine tumor, islet cell tumor, papillomatosis, paraganglioma, carcinoma of nasal sinus and nasal cavity, parathyroid carcinoma, carcinoma of penis, carcinoma of pharynx and larynx, pituitary tumor, pleuropulmonary blastoma, primary peritoneal carcinoma, retinoblastoma, salivary gland tumor, sarcoma, Sézary syndrome, skin cancer, small cell lung cancer, small intestinal carcinoma, soft tissue sarcoma, squamous cell carcinoma, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, endometrium and uterine sarcoma, vaginal carcinoma, vascular tumor, vulvar carcinoma, solitary myeloma, liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer and gastric cancer.

25. The method of claim 11, wherein the tumor is selected from a group consisting of: liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer and gastric cancer.

26. The method of claim 11, wherein the tumor is selected from a group consisting of: acute lymphoblastic leukemia, acute myelocytic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, lymphoma or multiple myeloma.

27. The method of claim 11, wherein the tumor is insensitive to alphavirus.

28. The method of claim 11, wherein the ratio of said proteasome inhibitor and said alphavirus is 0.01 to 200 mg: 10.sup.3 to 10.sup.9 PFU.

29. The method of claim 11, wherein 0.01 mg/kg to 200 mg/kg of said proteasome inhibitor is administered; and a titer at MOI from 10.sup.3 to 10.sup.9 PFU/kg of said alphavirus is administered.

30. The method of claim 11, wherein said proteasome inhibitor is administered intraperitoneally, intravenously, intra-arterially, intramuscularly, intradermally, intratumorally, subcutaneously or intranasally; and/or said alphavirus is administered intraperitoneally, intravenously, intra-arterially, intramuscularly, intradermally, intratumorally, subcutaneously or intranasally.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0087] FIG. 1 Bortezomib and M1 virus significantly increased morphological lesions of human hepatocellular carcinoma strains.

[0088] FIG. 2 Treatment of Bortezomib combined with M1 virus significantly reduced the viability of human hepatocellular carcinoma strains.

[0089] FIG. 3 Carfilzomib or treatment combined with M1 virus significantly inhibited the growth of human hepatocellular carcinoma strain xenograft tumor; wherein FIG. 3A is a flow chart of medicament treatment time; FIG. 3B shows that the treatment of Carfilzomib combined with M1 virus significantly inhibited the growth of human hepatocellular carcinoma strain Hep3B xenograft tumor; FIG. 3C shows that the treatment of Carfilzomib combined with M1 virus significantly inhibited the growth of human hepatocellular carcinoma strain Huh 7 xenograft tumor.

[0090] FIG. 4 Treatment of proteasome inhibitors combined with M1 virus significantly reduced the viability of human hepatocellular carcinoma strains; FIG. 4A shows that the treatment of CEP-18770 or combined with M1 virus significantly inhibits the viability of human hepatocellular carcinoma strains; FIG. 4B shows that MLN-9708 or treatment combined with M1 virus significantly inhibits the viability of human hepatocellular carcinoma strains; FIG. 4C shows that ONX-0912 or treatment combined with M1 virus significantly inhibited human hepatocellular carcinoma viability.

DETAILED DESCRIPTION

[0091] The following embodiments further illustrate the present invention, but the embodiments of the present invention are not limited to the following examples. Any equivalent changes or modifications made in accordance with the principles or concepts of the present invention should be regarded as the scope of protection of the present invention.

[0092] Without being specifically indicated, the materials and experimental methods employed in the present invention are conventional materials and methods.

[0093] The term “selected from” in the specification is used in connection with a selected object and is to be understood as, for example: “X is selected from: A, B, C, . . . , E” or “X is selected from one or more of A, B, C, . . . and E”, and the like, are understood to mean that X includes one of A, B, C, E, or any combination of both, or any combination of more. At this time, it is not excluded that X also includes some other types of substances.

[0094] In the present invention, the singular forms “a, an”, and “the” include plural form unless the context clearly dictates otherwise.

[0095] In the present invention, “treat” refers to alleviation of symptoms, temporary or permanent elimination of the cause of symptoms, or prevention or alleviation of the symptoms of a given disease or disorder.

[0096] In the present invention, “pharmaceutically acceptable carrier” refers to molecular entities and compositions that do not produce an allergic or similar adverse reaction when administered to a human. Including any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, the use thereof in the therapeutic compositions is contemplated.

[0097] In the present invention, a “pharmaceutically acceptable salt” is prepared by reacting the free acid or base form of a compound with a suitable base or acid in water or in an organic solvent or in a mixture of the two. Including acid addition salts or base addition salts. Examples of acid addition salts include inorganic acid addition salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, and organic acid addition salts such as acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate, and p-toluene sulfonate. Examples of base addition salts include inorganic salts such as sodium salt, potassium salt, calcium salt and ammonium salt, and organic base salts.

[0098] The term “effective amount” includes an amount of an alphavirus or proteasome inhibitor used in the present invention sufficient to provide the desired therapeutic effect. The exact amount required will vary from subject to subject, depending on factors such as: the species being treated, the age and general condition of the subject, the severity of the condition being treated, the particular agent being administered and the mode of administration, and the like. However, for a given situation, the dosage of the pharmaceutical compositions of the present invention may be adjusted by an ordinary skill in the art according to the severity of the symptoms, the frequency of recurrence, and the physiological response of the treatment regimen.

[0099] In addition to the above-mentioned proteasome inhibitors, the proteasome inhibitors of the present invention may be selected from proteasome inhibitors already known in the art, or substances found to have proteasome inhibition effect through subsequent studies. Examples of proteasome inhibitors include, but are not limited to, the following groups: Bortezomib, Carfilzomib, MG-132, ONX-0914, ONX-0912 (Oprozomib), CEP-18770 (Delanzomib), MLN-9708 (Ixazomib), Epoxomicin, VR23, MLN-2238, Celastrol, PI-1840, [(1R)-1-({[(2, 3-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2, 3-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(5-chloro-2-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3, 5-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2, 5-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2-bromobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2-chloro-5-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(4-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3, 4-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3-chlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2, 5-dichlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3, 4-dichlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2-chloro-4-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2, 3-dichlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(2-chlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boric acid, [(1R)-1-({[(2, 4-difluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boric acid, [(1R)-1-({[(4-chloro-2-fluorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boric acid, [(1R)-1-({[(4-chlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boric acid, [(1R)-1-({[(2, 4-dichlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, [(1R)-1-({[(3, 5-dichlorobenzoyl) amino] acetyl} amino)-3-methylbutyl] boronic acid, and mannitol esters, salts thereof or boronic acid anhydrides: which can also be found in patents WO07017440, EP11195107, U.S. 60/683,385, U.S. Ser. No. 09/300,779A, U.S. 60/815,218, WO04026407, and U.S. 60/495,764, all of which are incorporated herein by reference.

[0100] Depending on the type of tumor and the stage of disease progression, the effects of the tumor treatment methods of the present invention include, but are not limited to, inhibiting tumor growth, delaying tumor growth, tumor regression, tumor contraction, and increasing regeneration time of tumor when the treatment stops, slowing down the progression of the disease and preventing metastasis.

Example 1 Bortezomib and M1 Viruses Significantly Increased Morphological Lesions in Human Hepatoma Cell Strains

[0101] Materials:

[0102] Human hepatocellular carcinoma Hep3B (purchased from ATCC) and Huh 7 (purchased from ATCC), M1 virus (Accession No. CCTCC V201423), high glucose DMEM medium (purchased from Corning), inverted phase contrast microscope.

[0103] Method:

[0104] a) Cell culture: human hepatocellular carcinoma Hep3B and Huh 7 were grown in DMEM complete medium containing 10% FBS, 100 U/ml penicillin and 0.1 mg/ml streptomycin; all cell strains were cultured and passaged in a closed incubator at a constant temperature of 37° C. (95% relative humidity) with 5% CO.sub.2, and the growth was observed under inverted microscope. The cells were passaged about every 2 to 3 days, and the cells in the logarithmic phase were used in the formal experiment.

[0105] b) Cell treatment and morphological observation: cells in the logarithmic growth phase and the DMEM complete culture medium (containing 10% fetal bovine serum and 1% double antibody) were selected to prepare cell suspension, and the cells were inoculated in a 24-well culture plate at a density of 2.5×10.sup.4/well. The cells were treated with Bortezomib (5 nM) alone, infected with M1 virus (Hep3B: 0.001 moi, Huh 7: 0.1 moi) and treated with M1 virus combined with Bortezomib, with neither M1 virus nor Bortezomib as control, the morphological changes of cells were observed under inverted phase contrast microscope after 48 h.

[0106] Results:

[0107] As shown in FIG. 1, the cell morphology was observed under a phase contrast microscope, the Hep3B cells and Huh 7 cells in the control group grew in a monolayer with close arrangement and consistent phenotype, and there was no significant change in cell morphology after 48 h of treatment with Bortezomib (5 nM) or M1 virus (Hep3B: 0.001 moi, Huh 7: 0.1 moi), respectively. However, after 48 h of treatment by Bortezomib combined with M1 virus, compared with the control group and each single treatment group, the number of cells in the combined treatment group decreased significantly, and the morphology of the cells changed significantly, the cell body contracted into a ball, and the refractive index increased significantly, showing a death lesion.

Example 2 Treatment of Bortezomib Combined with M1 Virus Significantly Reduced the Viability of Human Hepatoma Cell Strains

[0108] Materials:

[0109] Human hepatocellular carcinoma Huh 7 (purchased from ATCC), M1 virus (Accession No. CCTCC V201423), high glucose DMEM medium (purchased from Corning), automatic enzyme-linked detection microplate reader.

[0110] Method:

[0111] a) Inoculating cells, and drug administering treatment: cells in the logarithmic growth phase and the DMEM complete culture medium (containing 10% fetal bovine serum and 1% double antibody) were selected to prepare cell suspension, and were inoculated in a 96-well culture plate at a density of 4×10.sup.3/well. 12 h later, the cells were completely adhered to the wall. The cells were divided into control group without medicament nor virus treatment, Bortezomib treatment group, M1 infection group and Bortezomib/M1 combined group. The doses used were as follows: M1 virus (MOI=0.001, 0.01, 0.1, 1, 10) infected cells, Bortezomib was 5 nM.

[0112] b) MTT reacted with intracellular succinate dehydrogenase: after culturing for 48 h, 20 μl (5 mg/ml) of MTT was added to each well and incubated for 4 h, at this time, the granular bluish violet formazan crystals formed in living cells could be observed under microscope.

[0113] c) Dissolving formazan granules: the supernatant was carefully removed, DMSO 100 μl/well was added to dissolve the crystal, shaked in a microoscillator for 5 min, and then detect the optical density (OD value) of each well with wavelength of 570 nm on the enzyme-linked detector. The experiment was repeated for 3 times in each group. Cell viability=OD value of medicament treatment group/OD value of control group×100%.

[0114] d) Origin 8 was used for nonlinear curve fitting, and two dose-response curves were drawn with drug dose as abscissa and relative cell viability as ordinate, namely, the dose-response curve of M1 virus alone and the dose-response curve of Bortezomib combined with M1 virus. The EC50 shift of the two curves was calculated, that is, the EC50 shift in FIG. 2, the larger the difference value was, the more significant the medicament synergized.

[0115] Results:

[0116] As shown in FIG. 2, treatment with Bortezomib (5 nM) alone had a small inhibitory effect on the viability of tumor cells Huh 7, and the relative cell viability of tumor cells reached 99.7%, the relative cell viability of the group treated with M1 virus (MOI=0.1) was still as high as 78.7%. However, when the same 5 nM Bortezomib was combined with M1 virus (MOI=0.1) (Eeyarestatin I+M1), the relative cell viability of tumor cells decreased significantly to 35.7%. Compared with single treatment, different doses of M1 virus (MOI=0.001, 0.01, 0.1, 1, 10) combined respectively with Bortezomib (5 nM) significantly decreased the viability of tumor cell Huh 7.

Example 3 Carfilzomib Combined with M1 Virus Significantly Inhibited the Growth of Human Hepatoma Cell Strain Xenograft Tumor

[0117] Materials:

[0118] M1 virus (accession number CCTCC V201423), human hepatoma cell strain Hep3B (purchased from ATCC), human hepatoma cell strain Huh 7 (purchased from ATCC), 4-week-old female BALB/c nude mice.

[0119] Method:

[0120] This experiment adopts a random, single-blind design. 5×10.sup.6 Hep 3B or Huh 7 cells were injected subcutaneously into the dorsal side of 4-week-old BALB/c nude mice. When the tumor size reached 50 mm.sup.3, the mice were grouped including untreated control group, Carfilzomib group (intraperitoneal injection 0.5 mg/kg/d), M1 infected group (tail vein injection of M1 virus 5×10.sup.5 PFU per time) and Carfilzomib/M1 combined group (the same dose of Carfilzomib and M1 virus was given in the same way), which were injected consecutively for 4 times in four days (see FIG. 3A). The length, width and body weight of the tumor were measured every two days, and the volume of the tumor was according to the formula (length×width 2)/2.

[0121] Results:

[0122] In two kinds of tumor cells (human hepatoma cell strain Hep3B and human hepatoma cell strain Huh 7) xenograft tumor animals, pathological anatomic measurements of tumor volume showed that, compared with the control group, Carfilzomib group and M1 infected group could only cause a slight reduction in tumor volume, while Carfilzomib/M1 combined group could cause a significant reduction in tumor volume (FIGS. 3B and 3C). At the end of the experiment, in the human hepatoma cell strain Hep3B model, the tumor volume of the control group was 2772.5 mm.sup.2, the tumor volume of Carfilzomib group and M1 infected group were 1668.5 mm.sup.2 and 1940 mm.sup.2, while the tumor volume of Carfilzomib/M1 combined group was 499 mm.sup.2. In the human hepatoma cell strain Huh 7 model, the tumor volume of the control group was 983.5 mm.sup.2, the tumor volume of the Carfilzomib group and the M1 infected group were 830.5 mm.sup.2 and 667.0 mm.sup.2, while the tumor volume of the Carfilzomib/M1 combined group was 313.7 mm.sup.2. One way ANOVA statistics showed that the difference was statistically significant (FIGS. 3B and 3C).

Example 4 Treatment of Various Proteasome Inhibitors Combined with M1 Virus Significantly Reduced the Viability of Human Hepatoma Cell Strains

[0123] Materials:

[0124] Human hepatocellular carcinoma Huh 7 (purchased from ATCC), M1 virus (Accession No. CCTCC V201423), high glucose DMEM medium (purchased from Corning), automatic enzyme-linked detection microplate reader.

[0125] Method:

[0126] a) Inoculating cells, and drug administering treatment: cells in the logarithmic growth phase and the DMEM complete culture medium (containing 10% fetal bovine serum and 1% double antibody) were selected to prepare cell suspension, and were inoculated in a 96-well culture plate at a density of 4×10.sup.3/well. 12 h later, the cells were completely adhered to the wall. The cells were divided into control group without medicament nor virus treatment, proteasome inhibitor group (including CEP-18770, MLN-9708, ONX-0912), M1 infected group and proteasome inhibitor/M1 combined group. The doses used were as follows: the doses used were as follows: M1 virus (MOI=0.1) infected cells; Proteasome inhibitor doses are as follows: CEP-18770 (5 nM), MLN-9708 (5 nM), ONX-0912 (50 nM).

[0127] b) MTT reacted with intracellular succinate dehydrogenase: after culturing for 72 h, 20 μl (5 mg/ml) of MTT was added to each well and incubated for 4 h, at this time, the granular bluish violet formazan crystals formed in living cells could be observed under microscope.

[0128] c) Dissolving formazan granules: the supernatant was carefully removed, DMSO 100 μl/well was added to dissolve the crystal, vibrate in a microoscillator for 5 min, and then detect the optical density (OD value) of each well with wavelength of 570 nm on the enzyme-linked detector. Cell viability=OD value of medicament treatment group/OD value of control group x 100%.

[0129] Results:

[0130] As shown in FIG. 4A, treatment with CEP-18770 alone had a small effect on the viability of tumor cells Huh 7, and the relative cell viability of tumor cells reached 105.4%, the relative cell viability of the group treated with M1 virus (MOI=0.1) was still as high as 84.6%. However, when the same CEP-18770 was combined with M1 virus (MOI=0.1), the relative cell viability of tumor cells decreased significantly to 42.2%; as shown in FIG. 4B, treatment with MLN-9708 alone had a small effect on the viability of tumor cells Huh 7, and the relative cell viability of tumor cells reached 77.7%, the relative cell viability of the group treated with M1 virus (MOI=0.1) was still as high as 84.6%. However, when the same CEP-18770 was combined with M1 virus (MOI=0.1), the relative cell viability of tumor cells decreased significantly to 45.3%; as shown in FIG. 4C, treatment with ONX-0912 alone had a small effect on the viability of tumor cells Huh 7, and the relative cell viability of tumor cells reached 70.0%, the relative cell viability of the group treated with M1 virus (MOI=0.1) was still as high as 84.6%. However, when the same CEP-18770 was combined with M1 virus (MOI=0.1), the relative cell viability of tumor cells decreased significantly to 37.8%.

[0131] The described embodiments of the present invention are merely illustrative examples, and the embodiments of the present invention are not limited to the above, and any other changes, modifications, substitutions, combinations, and simplifications that may be made without departing from the spirit and principles of the present invention shall be equivalent replacement and shall be included in the protection scope of the present invention.