Medicament for treatment and / or prevention of cancer

Abstract

A combined medicament that comprises particles containing an amphiphilic polymer, in which the hydrophobic segment is a poly(hydroxy acid) and the hydrophilic segment is a polysaccharide, and a cancer antigen with an immune-activating factor. By using the combined medicament in such a manner that the particles and the immune-activating factor are administered separately to a subject, the effectiveness of cancer treatment and / or prevention can be enhanced.

Description

Medicines for the treatment and / or prevention of cancer

[0001] The present invention relates to a pharmaceutical for treating and / or preventing cancer, which comprises administering a cancer antigen and an immune activator separately.

[0002] As a method of treating or preventing cancer, attempts have been made to develop "cancer vaccines" that induce an immune response against cancer cells in the body by administering cancer antigens (proteins, their peptide fragments, etc.) that are specifically expressed in cancer cells. Unlike typical low-molecular-weight anticancer drugs, only cancer cells are attacked by immune cells, resulting in fewer side effects. Furthermore, by establishing immune memory in the body, they are expected to be effective in preventing recurrence. However, due to the high difficulty of technological development, only a few cancer vaccines have been launched on the market, such as "Provenge" (a dendritic cell vaccine for prostate cancer) and "Imygic" (an oncolytic virus preparation for metastatic melanoma).

[0003] The development of powerful adjuvants is needed to improve immune responses to antigens. However, due to safety concerns, only a limited number of adjuvants are currently available for human use, and no substances with reported high adjuvant effects have been approved. For example, aluminum hydroxide adjuvants have limited immune activation potential, requiring repeated administration to achieve immunity.

[0004] As a new adjuvant aimed at achieving high immune activation, methods of encapsulating antigens in particles have been attempted. This method is expected to improve safety and delivery efficiency to target cells by encapsulating antigens through particle formation. Particulate adjuvants have been proposed using a wide variety of materials, including fatty acids, biodegradable polymers, and viral proteins (Non-Patent Documents 1 and 2). Recently, technology has been reported relating to antigen-adjuvant particle complexes in which antigens are encapsulated in adjuvant particles composed of amphiphilic polymers whose hydrophobic segments are poly(hydroxy acid)s and whose hydrophilic segments are polysaccharides (Patent Documents 1 and 2). In particular, Patent Document 2 describes the use of complexes with antigens expressed in cancer, demonstrating antitumor effects, and the application of particulate adjuvants as cancer therapeutics is anticipated.

[0005] In clinical practice, a treatment that combines multiple cancer therapeutic drugs is used as a standard treatment to enhance the effectiveness of cancer therapeutic drugs. Regarding the above-mentioned particulate adjuvant, Patent Document 1 describes that its immune activation ability is enhanced by mixing it with an immune activating factor.

[0006] WO2010 / 098432WO2015 / 053354

[0007] Immunology, 2006, No. 117, pp. 78-88 Nature Materials, 2011, No. 10, pp. 243-251

[0008] The combined use of particulate adjuvants and immune activators is promising as a cancer treatment or prevention method, but there has been insufficient research into effective combinations. Therefore, an objective of the present invention is to establish a technology that enhances the cancer treatment and prevention effects of pharmaceuticals that combine particulate adjuvants and immune activators.

[0009] In order to overcome the above-mentioned problems, the present inventors have conducted extensive research focusing on particles (hereinafter, in this specification, often referred to as "particles") containing an amphiphilic polymer, in which the hydrophobic segment is poly(hydroxy acid) and the hydrophilic segment is polysaccharide, and a cancer antigen, which are promising particulate adjuvants for the treatment or prevention of cancer. As a result, they have found that a significant antitumor effect can be achieved by administering the particles and an immune activator separately, and have completed the present invention.

[0010] That is, the present invention has the following configurations (1) to (15). (1) A pharmaceutical for treating and / or preventing cancer, characterized in that particles comprising an amphiphilic polymer whose hydrophobic segment is a poly(hydroxy acid) and a hydrophilic segment is a polysaccharide, a cancer antigen, and an immune activator are administered separately. (1-1) A combination pharmaceutical for treating and / or preventing cancer, comprising particles comprising an amphiphilic polymer whose hydrophobic segment is a poly(hydroxy acid) and a hydrophilic segment is a polysaccharide, a cancer antigen, and an immune activator, characterized in that the particles and the immune activator are administered separately. (1-2) The pharmaceutical according to (1-1), characterized in that the particles and the immune activator are administered separately using different administration methods. (2) The pharmaceutical according to any one of (1) to (1-2), wherein the immune activator is a ligand or agonist that binds to a Toll-like receptor (TLR), a NOD-like receptor (NLR), a RIG-like receptor, a C-type lectin receptor (CLR), or a stimulator of interferon genes (STING). (3) The pharmaceutical according to (2), wherein the ligand or agonist that binds to a Toll-like receptor (TLR) is a ligand or agonist that binds to TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, or TLR11. (4) The pharmaceutical according to (3), wherein the ligand or agonist that binds to a Toll-like receptor (TLR) is any one of the following (i) to (vii):(i) a ligand or agonist that binds to TLR2 selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide, and zymosan; (ii) a ligand or agonist that binds to TLR3 selected from the group consisting of Poly(I:C) and poly(A:U); (iii) a ligand or agonist that binds to TLR4 selected from the group consisting of lipopolysaccharide (LPS), HSP60, RS09, and MPLA; (iv) flagellin, which is a ligand or agonist that binds to TLR5; (v) a ligand or agonist that binds to TLR7 or 8 selected from the group consisting of imidazoquinoline compounds and single-stranded RNA; and (vi) a ligand or agonist that binds to TLR9 selected from the group consisting of bacterial DNA, unmethylated CpG DNA, hemozorin, ODN1585, ODN1668, and ODN1826. (vii) A ligand or agonist that binds to TLR11 selected from the group consisting of profilin and uropathogenic bacteria. (5) The pharmaceutical according to (4), wherein the imidazoquinoline compound is imiquimod. (6) The pharmaceutical according to any of (1) to (5), wherein the immune activator is administered locally to a tumor. (6-1) The pharmaceutical according to any of (1) to (5), wherein the immune activator is administered locally to a tumor. (7) The pharmaceutical according to any of (1) to (6-1), wherein the polysaccharide is dextran, β-glucan, mannan, chitin, chitosan, gellan gum, alginic acid, hyaluronic acid, or pullulan. (8) The pharmaceutical according to (7), wherein the β-glucan is a polymer of glucose linked by one or more β-1,3 bonds and / or one or more β-1,6 bonds. (9) The pharmaceutical according to (7) or (8), wherein the β-glucan is black yeast glucan, curdlan, pachyman, laminaran, lichenan, schizophyllan, lentinan, scleroglucan, or pachymaran. (10) The pharmaceutical according to any one of (1) to (9), wherein the poly(hydroxy acid) is poly(lactic acid-glycolic acid), polylactic acid, or polyglycolic acid. (11) The pharmaceutical according to any one of (1) to (10), wherein the number-average molecular weight of the amphiphilic polymer is 500 to 100,000.(12) The pharmaceutical according to any one of (1) to (11), wherein the particles have an average particle size of 0.1 to 50 μm. (13) The pharmaceutical according to any one of (1) to (12), wherein the cancer is basal cell carcinoma, Paget's disease, skin cancer, breast cancer, kidney cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cell tumor, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, or head and neck cancer. (14) A method for treating and / or preventing cancer, comprising separately administering particles comprising an amphiphilic polymer, the hydrophobic segment of which is a poly(hydroxy acid) and the hydrophilic segment is a polysaccharide, and a cancer antigen, and an immune activator. (15) The method according to (14), wherein the particles and the immunoactivator are administered separately using different administration methods.

[0011] This specification includes the disclosure of Japanese Patent Application No. 2021-150931, from which this application claims priority.

[0012] The use of the amphiphilic polymer of the present invention, in which the hydrophobic segment is a poly(hydroxy acid) and the hydrophilic segment is a polysaccharide, particles containing a cancer antigen, and an immune activator, administered separately, exhibits a significant antitumor effect and is effective in the treatment and prevention of cancer.

[0013] Dextran-NH ２ The results of GPC measurement of dextran-PLGA are shown below. １ The results of H-NMR measurement are shown below. ２ The results of GPC measurement of black yeast glucan-PLGA are shown. １ 1 shows the results of H-NMR measurement. 2 shows the results of DLS measurement of OVA-containing dextran particles. 3 shows the results of DLS measurement of OVA-containing black yeast glucan particles.

[0014] The present invention is characterized by inducing strong immune activation by using particles comprising an amphiphilic polymer, the hydrophobic segment of which is a poly(hydroxy acid) and the hydrophilic segment of which is a polysaccharide, a cancer antigen, and an immune activator, which are administered separately. The type of immune response induced by the pharmaceutical of the present invention is not limited. Types of immune responses include Th1-type immune responses and Th2-type immune responses, and it is known that one type of immune response is predominantly induced depending on the antigen, the administration site, the administration method, and the type of immune activator combined with the particles. However, the present invention can induce both Th1-type and Th2-type immune responses.

[0015] The antitumor activity of the particles containing an amphiphilic polymer, the hydrophobic segment of which is a poly(hydroxy acid) and the hydrophilic segment of which is a polysaccharide, and a cancer antigen used in the present invention in combination with an immune activator can be evaluated by examining the inhibition of tumor growth in tumor-bearing animals in vivo, as described below.

[0016] Among the pharmaceuticals of the present invention, the amphiphilic polymer that constitutes the particles together with the cancer antigen will be described below. "Amphiphilic" means having both hydrophilic and hydrophobic properties. When a certain portion (segment) has higher solubility in water than other portions (segments), that portion is said to be hydrophilic. The hydrophilic portion is preferably soluble in water, but even if it is sparingly soluble, it is acceptable as long as it has higher solubility in water than other portions. Furthermore, when the solubility in water of a certain portion (segment) is lower than other portions, that portion (segment) is said to be hydrophobic. The hydrophobic portion is preferably insoluble in water, but even if it is soluble, it is acceptable as long as it has lower solubility in water than other portions.

[0017] An "amphiphilic polymer" is a polymer that has the above-described amphiphilic properties as a whole molecule. An amphiphilic "polymer" refers to a molecular structure in which the hydrophilic segment, the hydrophobic segment, or both in the amphiphilic molecule are composed of repeating structures of minimum units (monomers). The structure of the amphiphilic polymer in the present invention is not particularly limited, and specific examples include linear block polymers in which polysaccharides and poly(hydroxy acids) are linked together, branched polymers having multiple branches containing polysaccharides or poly(hydroxy acids), graft polymers consisting of a polysaccharide main chain and a poly(hydroxy acid) side chain, and graft polymers consisting of a poly(hydroxy acid) main chain and a polysaccharide side chain. A linear block polymer in which polysaccharides and poly(hydroxy acids) are linked together is preferred.

[0018] The present invention is characterized in that the hydrophilic segment of the amphiphilic polymer is a polysaccharide. While there are no particular limitations on the polysaccharide, specific examples include dextran, β-glucan, mannan, chitin, chitosan, gellan gum, alginic acid, hyaluronic acid, and pullulan, and dextran or β-glucan is preferred.

[0019] Glucan is a glucose-containing polysaccharide, and β-glucan contains one or more β-bonds between glucose subunits. That is, the β-glucan used in the present invention contains β-bonds, or may contain only β-bonds. Furthermore, the β-glucan used in the present invention may be branched or linear. Preferred β-glucans include those containing one or more β-1,3 bonds and / or one or more β-1,6 bonds, and those containing one or more β-1,2 bonds and / or β-1,4 bonds. However, those containing one or more β-1,3 bonds and / or one or more β-1,6 bonds are more preferred, and those containing one or more β-1,3 bonds are even more preferred. Specific examples of β-glucans containing one or more β-1,3 bonds include curdlan, pachyman, laminaran, lichenan, schizophyllan, lentinan, scleroglucan, black yeast glucan (preferably, β-1,3 glucan or β-1,6 glucan derived from black yeast), or pachymaran, and preferred examples include curdlan, pachyman, laminaran, schizophyllan, scleroglucan, black yeast glucan, or pachymaran.

[0020] Examples of linear β-glucans containing one or more β-1,3 bonds include β-glucans consisting mainly of β-1,3 bonds (e.g., curdlan and pachyman) and β-glucans consisting of β-1,3 bonds and β-bonds other than β-1,3 bonds (e.g., laminaran and lichenan).

[0021] Examples of branched β-glucans containing one or more β-1,3 bonds include β-glucans consisting of β-1,3 and β-1,6 bonds (for example, schizophyllan, lentinan, scleroglucan, and black yeast glucan).

[0022] The β-glucan used in the present invention may be a derivatized β-glucan. Examples of derivatization include an addition reaction of a carboxymethyl group and an oxidative cleavage reaction. Examples of derivatized β-glucans include carboxymethylcurdlan, in which a carboxymethyl group is added to curdlan, and pachymaran, in which pachyman is cleaved.

[0023] The number average molecular weight of the polysaccharide is not particularly limited, but is preferably 500 to 100,000, more preferably 500 to 50,000, and even more preferably 1,000 to 10,000, for example, 1,000 to 9,000, 1,000 to 8,000, 1,000 to 7,000, 1,000 to 6,000, 1,000 to 5,000, 1,000 to 4,000, or 1,000 to 3,000. The number average molecular weight is an average molecular weight calculated by a method that does not take into account weighting of molecular size (simple average), and the number average molecular weight of the polysaccharide can be determined by gel permeation chromatography (GPC).

[0024] In the present invention, the hydrophobic segment of the amphiphilic polymer is characterized by being a poly(hydroxy acid). While the poly(hydroxy acid) is not particularly limited, it is preferably a biocompatible polymer that does not cause significant adverse effects when administered to a living body. Biocompatibility here refers to a polymer with an LD50 of 2,000 mg / kg or greater when orally administered to rats. The poly(hydroxy acid) may also be a copolymer of multiple types of hydroxy acids, but is preferably a polymer of two or fewer types of hydroxy acids.

[0025] Specific preferred examples of poly(hydroxy acids) include polyglycolic acid, polylactic acid, poly(2-hydroxybutyric acid), poly(2-hydroxyvaleric acid), poly(2-hydroxycaproic acid), poly(2-hydroxycapric acid), poly(malic acid), and derivatives and copolymers of these polymer compounds, with poly(lactic acid-glycolic acid), polylactic acid, or polyglycolic acid being preferred, and poly(lactic acid-glycolic acid) being more preferred. Furthermore, when the poly(hydroxy acid) is poly(lactic acid-glycolic acid), the composition ratio (lactic acid / glycolic acid) (mol / mol) of the poly(lactic acid-glycolic acid) is not particularly limited as long as the object of the present invention is achieved, but is preferably 99 / 1 to 1 / 99, and more preferably 80 / 20 to 20 / 80, e.g., 60 / 40 to 40 / 60, or 50 / 50.

[0026] The number average molecular weight of the poly(hydroxy acid) moiety in the amphiphilic polymer of the present invention is not particularly limited, but is preferably 500 to 1,000,000, more preferably 500 to 100,000, and even more preferably 7,000 to 50,000, 7,000 to 50,000, 7,000 to 40,000, 7,000 to 30,000, 7,000 to 20,000, 7,000 to 15,000, 7,000 to 13,000, or 7,000 to 12,000. The number average molecular weight of the poly(hydroxy acid) is determined from the difference between the number average molecular weight of an amphiphilic polymer in which the hydrophobic segment is a poly(hydroxy acid) and the hydrophilic segment is a polysaccharide and the number average molecular weight of the polysaccharide.

[0027] The number average molecular weight of the amphiphilic polymer constituting the particles is not particularly limited, but is preferably 1,000 to 1,000,000, more preferably 1,000 to 100,000, and even more preferably 9,000 to 50,000, 9,000 to 50,000, 9,000 to 40,000, 9,000 to 30,000, 9,000 to 20,000, or 9,000 to 15,000. The number average molecular weight of the amphiphilic polymer is determined by gel permeation chromatography (GPC).

[0028] The amphiphilic polymer may be produced by a known method, specifically, a method of adding a poly(hydroxy acid) to a polysaccharide and carrying out a condensation reaction, or a method of adding a hydroxy acid-activated monomer to a polysaccharide and carrying out a polymerization reaction, can be mentioned as examples.

[0029] Furthermore, when the amphiphilic polymer is a linear block polymer in which a polysaccharide and a poly(hydroxy acid) are linked together, it may be produced by a known method, specifically, a production method in which a poly(hydroxy acid) copolymer is subjected to a condensation reaction with the reducing end of the polysaccharide of the amphiphilic polymer using an activator for the terminal functional group [Macromol. Rapid Commun., 31, pp. 1664-1684 (2010)] can be mentioned as an example.

[0030] When the amphiphilic polymer is a graft polymer having a polysaccharide main chain and a poly(hydroxy acid) side chain, it can be produced as in the following (1), (2), or (3).

[0031] (1) A method for producing a graft polymer by adding a hydroxy acid-activated monomer to a polysaccharide in the presence of a tin catalyst to carry out a polymerization reaction and introducing poly(hydroxy acid) [Macromolecules, 31, pp. 1032-1039 (1998)]. (2) A method for producing a graft polymer by activating, with a base, the partially unprotected hydroxy groups of a polysaccharide, most of whose hydroxy groups are protected by substituents, followed by adding a hydroxy acid-activated monomer to introduce a graft chain consisting of poly(hydroxy acid), and finally removing the protecting groups [Polymer, 44, pp. 3927-3933 (2003)]. (3) A method for producing a graft polymer by condensing a poly(hydroxy acid) copolymer with a polysaccharide using a dehydrating agent and / or a functional group activator [Macromolecules, 33, pp. 3680-3685 (2000)].

[0032] In order to maintain the immunostimulatory effect over a long period of time, the amphiphilic polymer is preferably water-insoluble as a whole so that it is not immediately excreted from the body. "Water-insoluble" here means that the solubility in water is 1 g (amphiphilic polymer) / 100 ml (water) or less.

[0033] The amphiphilic polymer may be a modified amphiphilic polymer that has been modified with a functional compound to impart a desired function, such as a modified amphiphilic polymer that has been conjugated to an immunostimulatory factor, such as those described below, by a known method in order to enhance the immunostimulatory effect.

[0034] The cancer antigen that constitutes the particles together with the amphiphilic polymer will now be described. The "cancer antigen" of the present invention is an antigen that can be used as a medicine or vaccine for the treatment and / or prevention of cancer by eliciting an immune response in the body. By using particles containing the amphiphilic polymer and cancer antigen of the present invention as active ingredients, the immune response elicited by the cancer antigen can be enhanced.

[0035] The cancer antigen contained in the particles of the present invention is not particularly limited as long as the object of the present invention is achieved. Examples of cancer antigens include peptides, proteins, glycoproteins, glycolipids, lipids, carbohydrates, nucleic acids, polysaccharides, and viruses, bacteria, tissues, cells, etc. containing these. Specific examples include cancer antigens described in WO2017 / 181128, such as WT1, MUC1, LMP2, HPV E6, HPV E7, EGFRvIII, Her-2 / neu, idiotype, MAGE A3, p53, NY-ESO-1 (CTAG1), PSMA, CEA, MelanA / Mart1, Ras, gp100, proteinase 3, bcr-able, tyrosinase, survivin, PSA, hTERT, and sarcoma translocation breakpoints. breakpoints), EphA2, PAP, MP-IAP, AFP, EpCAM, ERG, NA17-A, PAX3, ALK, androgen receptor, cyclin B1, MYCN, PhoC, TRP-2, mesothelin, PSCA, MAGE A1, CYP1B1, PLAC1, BORIS, ETV6-AML, NY-BR-1, RGS5, SART3, carbonic anhydrase IX, PAX5, OY-TES1, sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE1, B7-H3, legumain, Tie2, Page4, VEGFR2, MAD-CT-1, FAP, PAP, PD Examples of such antigens include GFR-beta, MAD-CT-2, CEA, TRP-1 (gp75), BAGE1, BAGE2, BAGE3, BAGE4, ​​BAGE5, CAMEL, MAGE-A2, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, and Fos-related antigen 1. Furthermore, as a model experimental system for evaluating the enhancement of the antitumor effect of a cancer antigen by a particulate adjuvant, an experimental system in which a particulate model cancer antigen such as ovalbumin (OVA) is allowed to act on cancer cells in which the model cancer antigen has been forcibly expressed is sometimes used; the term "cancer antigen" as used herein also encompasses such model cancer antigens. The number of cancer antigens contained in the particles of the present invention is not particularly limited, and may be one type or two or more types.

[0036] The particles of the present invention, which are one of the active ingredients and are composed of an amphiphilic polymer and an antigen, may contain additional components other than the cancer antigen and amphiphilic polymer. Specific examples of additional components include lipids, which are well-known components of liposomes and lipid nanoparticles, which are particles that can encapsulate antigens, such as cancer antigens. However, as is clear from the examples, the present invention can achieve the expected effects without using lipids as components other than the cancer antigen that constitute the particles, so it is preferable that the particles do not contain lipids as additional components.

[0037] The structure of the particles composed of an amphiphilic polymer and a cancer antigen, which is one of the active ingredients of the present invention, is not particularly limited. However, since the particles are complexes of an amphiphilic polymer and a cancer antigen, it is preferable to adopt a structure in which the hydrophilic segment of the amphiphilic polymer constituting the particles is on the inside of the particle and the hydrophobic segment is on the outer layer of the particle, because this allows the cancer antigen to be encapsulated in the particle and be more stably retained.

[0038] The method for producing the particles is not particularly limited, and examples thereof include a submerged drying method, a spray drying method, and a pulverization method. The particles of the present invention are preferably produced by a submerged drying method.

[0039] Examples of methods for producing particles by the submerged drying method include an O / W emulsion method, a W / O / W emulsion method, and an S / O / W emulsion method.

[0040] For example, when particles are produced by the O / W emulsion method, they can be produced by mixing a water-immiscible organic solvent in which a powder of an amphiphilic polymer constituting the particles has been dissolved with an aqueous solution in which a surface modifier and a cancer antigen have been dissolved to prepare an O / W emulsion solution, and then removing the water-immiscible organic solvent from the O / W emulsion solution to obtain particles.

[0041] An example of producing particles by the W / O / W emulsion method includes the steps of: mixing an aqueous solvent in which a cancer antigen has been dissolved with a water-immiscible organic solvent in which an amphiphilic polymer powder constituting the particles has been dissolved to prepare a W / O emulsion solution; mixing the W / O emulsion solution with an aqueous solution of a surface modifier to prepare a W / O / W emulsion solution; and removing the water-immiscible organic solvent from the W / O / W emulsion solution to obtain particles.

[0042] An example of producing particles by the S / O / W emulsion method includes the steps of: mixing an aqueous solvent in which a cancer antigen has been dissolved with a water-immiscible organic solvent in which an amphiphilic polymer powder constituting the particles has been dissolved to prepare a W / O emulsion solution; removing the solvent from the W / O emulsion solution to obtain a solid; dispersing the solid in a water-immiscible organic solvent to obtain an S / O suspension solution; mixing the S / O suspension solution with an aqueous solution of a surface modifier to prepare an S / O / W emulsion solution; and removing the water-immiscible organic solvent from the S / O / W emulsion solution to obtain particles.

[0043] The antigen content in the particles (antigen / particle) is preferably 0.01 to 20% by weight, more preferably 0.1 to 10% by weight. The antigen content can be determined by extracting the antigen from the particles using an organic solvent and quantifying the antigen by gel electrophoresis or liquid chromatography.

[0044] The surface modifier used for particle preparation is preferably a water-soluble polymer or a surfactant. The water-soluble polymer here refers to a polymer compound having a solubility in water of 1 g (water-soluble polymer) / 100 ml (water) or more.

[0045] Examples of water-soluble polymers that can be used as surface modifiers include polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, polyethyleneimine, polyacrylic acid, polymethacrylic acid, poly-1,3-dioxolane, 2-methacryloyloxyethyl phosphorylcholine polymer, poly-1,3,6-trioxane, polyamino acids, peptides, proteins, and sugars (monosaccharides, oligosaccharides, polysaccharides, etc.), with polyvinyl alcohol being preferred.

[0046] Examples of surfactants that can be used as surface modifiers include nonionic surfactants such as polyoxyethylene polyoxypropylene glycol, sucrose fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan mono-fatty acid esters, polyoxyethylene sorbitan di-fatty acid esters, polyoxyethylene glycerin mono-fatty acid esters, polyoxyethylene glycerin di-fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene castor oil, and polyoxyethylene hydrogenated castor oil; alkyl sulfates such as sodium lauryl sulfate, ammonium lauryl sulfate, and sodium stearyl sulfate; and lecithin, with polyoxyethylene polyoxypropylene glycol being preferred.

[0047] The water-immiscible organic solvent used for particle preparation is preferably one in which the amphiphilic polymer is soluble and the polysaccharide is poorly soluble or insoluble. The solubility of the water-immiscible organic solvent in water is preferably 30 g (water-immiscible organic solvent) / 100 ml (water) or less. Specific examples of water-immiscible organic solvents include ethyl acetate, isopropyl acetate, butyl acetate, dimethyl carbonate, diethyl carbonate, methylene chloride, and chloroform.

[0048] The aqueous solvent used for particle preparation is an aqueous solution containing water and, optionally, water-soluble components, such as inorganic salts, sugars, organic salts, amino acids, peptides, proteins, and nucleic acids.

[0049] The particle surface may be bonded to a surface modifier used in the manufacturing process. The bond here may be a non-covalent bond or a covalent bond. The non-covalent bond is preferably a hydrophobic interaction, but may also be an ionic bond (electrostatic interaction), a hydrogen bond, a coordinate bond, a van der Waals bond, a physical adsorption, or a combination thereof.

[0050] The average particle size of the particles is preferably 0.1 μm to 50 μm, more preferably 0.1 μm to 25 μm, even more preferably 0.1 μm to 10 μm, and particularly preferably 0.1 μm to 1 μm, for example, 0.2 μm to 0.9 μm, 0.3 μm to 0.8 μm, or 0.4 μm to 0.7 μm. The average particle size here can be determined by the cumulant method using a dynamic light scattering device (DLS: for example, ELS-Z, manufactured by Otsuka Electronics Co., Ltd.).

[0051] As used herein, "treatment" refers to the treatment of cancer based on the antitumor effect described above. Furthermore, as used herein, "prevention" refers not only to the prevention of cancer onset, but also to the prevention of cancer metastasis or recurrence.

[0052] As used herein, the terms "tumor" and "cancer" refer to malignant neoplasms and are used interchangeably.

[0053] There are no particular limitations on the method of administering the particles of the pharmaceutical of the present invention to a living body (subject), but examples include subcutaneous administration, intradermal administration, intramuscular administration, nasal administration, pulmonary administration, oral administration, transdermal administration, sublingual administration, intravaginal administration, intraperitoneal administration, and lymph node administration, with intradermal or subcutaneous administration being preferred.

[0054] The method of administering the particles is preferably selected to be different from the method of administering the immunoactivator described below. For example, when the immunoactivator is administered locally to a tumor, the particles can be administered subcutaneously.

[0055] The dosage of the particles is determined appropriately depending on the administration method and frequency of administration. For example, when the particles of the present invention are administered subcutaneously to humans, the dosage can be selected from the range of 0.0001 mg to 1,000 mg per kg of body weight per administration. Alternatively, the dosage can be selected from the range of 0.001 mg / body to 100,000 mg / body per patient, or from 1 mg to 30 mg per kg of patient body weight, but is not necessarily limited to these values. The dosage and administration method vary depending on the patient's body weight, age, sex, symptoms, etc., but can be appropriately selected by one skilled in the art.

[0056] The immune activator, which is one of the active ingredients of the present invention, will now be described. The term "immune activator" as used herein refers to a factor that activates one or more types of immune cells and is a substance that can maintain or enhance the immune function of the cells. The term "immune cells" as used herein refers to, for example, T lymphocytes, B lymphocytes, NK cells, monocytes, dendritic cells, granulocytes, macrophages, myeloid-derived suppressor cells, Langerhans cells and their precursor cells, and the immune cell groups present in tumors.

[0057] The immune activator used in the present invention is not particularly limited, and specific examples include ligands or agonists that bind to Toll-like receptors (TLRs), NOD-like receptors (NLRs), RIG-like receptors, C-type lectin receptors (CLRs), or stimulatory factor for interferon genes (STINGs), with preference given to ligands or agonists that bind to TLRs.

[0058] Specific examples of TLRs include TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, and TLR11, and specific examples of ligands or agonists that bind to them include the following (i) to (vii): (i) a ligand or agonist that binds to TLR2 selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide, and zymosan; (ii) a ligand or agonist that binds to TLR3 selected from the group consisting of poly(I:C) and poly(A:U); (iii) a ligand or agonist that binds to TLR4 selected from the group consisting of lipopolysaccharide (LPS), HSP60, RS09, and MPLA; and (iv) flagellin, which is a ligand or agonist that binds to TLR5. (v) a ligand or agonist that binds to TLR7 or 8 selected from the group consisting of imidazoquinoline compounds and single-stranded RNA, (vi) a ligand or agonist that binds to TLR9 selected from the group consisting of bacterial DNA, unmethylated CpG DNA, hemozorin, ODN1585, ODN1668, and ODN1826, and (vii) a ligand or agonist that binds to TLR11 selected from the group consisting of profilin and uropathogenic bacteria.

[0059] In the present invention, the immunoactivator is preferably a ligand or agonist that binds to TLR3, or a ligand or agonist that binds to TLR7 and / or TLR8, and more preferably a ligand or agonist of (ii) or (v) below.

[0060] The (ii) TLR3-binding ligand or agonist selected from the group consisting of Poly(I:C) and Poly(A:U) is preferably Poly(I:C). Poly(I:C) is a double-stranded RNA formed by an RNA strand consisting only of inosine bases and an RNA strand consisting only of cytidine bases. The chain length of Poly(I:C) used in the present invention is not particularly limited.

[0061] The imidazoquinoline compound (v) is not particularly limited as long as it binds to TLR7 or TLR8. Preferred specific examples include the compounds described in U.S. Patent No. 8,951,528 and WO2015 / 103989, such as 4-amino-2-(ethoxymethyl)-a,a-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol (Resiquimod (R848)), 1-(2-methylpropyl)-1H- imidazo[4,5-c]quinoline-4-amine (Imiquimod), 1-(4-amino-2-eth ylaminomethylimidazo-[4,5-c]quinolin-1-yl)-2-methylpropan-2 -ol(Gardiquimod), N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]qui nolin-1-yl)butyl-]methanesulfonamide (PF-4878691), 4-amino-aa -dimethyl-2-methoxyethyl-1H-imidazo[4,5-c]quinoline-1-ethan ol, 1-(2-(3-(benzyloxy)propoxy)ethyl)-2-(ethoxymethyl)-1H-im idazo[4,5-c]quinolin-4-amine, 4-amino-2-ethoxymethyl-aa-dime thyl-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinoline-1-ethano l, N-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quin olin-l-yl]eyhoxy}ethyl)-n'-phenylurea, 1-2-amino-2-methylpro pyl)-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-4-amine, 1-{ 4-[(3,5-dichlorophenyl)sulfonyl]butyl}-2-ethyl-1H-imidazo[4,5-c] quinolin-4-amine, N-(2-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}eth yl)-n'-cyclohexylurea, N-{3-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]propyl}-n'-( Examples of the imiquimod are N-[3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-l-yl)-2,2-dimethylpropyl]benzamide, 2-butyl-1-[3-(methylsulfonyl)propyl]-1H-imidazo[4,5-c]quinolin-4-amine, and derivatives thereof. Imiquimod is more preferred, and imiquimod is a compound with a molecular weight of approximately 240.3 and represented by CAS number 99011-02-6. The IUPAC name for imiquimod is 4-amino-1-(2-methylpropyl)-1H-imidazo[4,5-c]quinoline, and other names include R837, S-26308, 1-(2-METHYLPROPYL)-1H-IMIDAZO[4,5-C]QUINOLIN-4-AMINE; 4-am It is represented by ino-1-isobutyl-1h-imidazo[4,5-c]quinolinone, 1-isobutyl-1H-inudazole[4.5-c]quinolin-4-amine; 1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine.

[0062] Specific examples of ligands or agonists that bind to NOD-like receptors (NLRs) include M-TriDAP and PGN. Other examples include ligands or agonists for NOD1, such as Tri-DAP, iE-DAP, and C12-iE. Furthermore, other examples include ligands or agonists for NOD2, such as MDP, N-glycosyl-MDP, Murabutide, M-TriLyS-D-ASN, M-TriLYS, and L18-MDP.

[0063] Specific examples of ligands or agonists that bind to RIG-like receptors include 5'ppp-dsRNA, poly(dA:dT), poly(dG:dC), and poly(I:C).

[0064] Specific examples of ligands or agonists that bind to C-type lectin receptors (CLR) include trehalose 6,6-dibehenate, zymosan, WGP, HKSC, HKCA, and curdlan AL.

[0065] Ligands or agonists that bind to stimulator of interferon genes (STING) include c-di-GMP, c-di-AMP, 2'3'-cGAMP, 3'3'-cGAMP, or 2'2'-cGAMP.

[0066] Imiquimod may be obtained by chemical synthesis using methods known to those skilled in the art, or commercially available pharmaceuticals may be used. The dosage form of imiquimod when administered to a patient is preferably a transdermal administration type, preferably a cream-type preparation. Pharmaceuticals containing imiquimod as an active ingredient include "Beselna Cream 5%" in Japan and "Aldara (registered trademark) Cream, 5%" in Europe and the United States. When using imiquimod in the present invention, these pharmaceuticals can be administered according to the dosage and administration instructions described in the package insert.

[0067] As used herein, the terms "coadministration" and "combination" refer to the administration of the particles and the immunoactivator to the same organism simultaneously or at a predetermined interval, each as an independent formulation (i.e., separately). The interval between the administration of one and the other is not particularly limited. The interval may be simultaneous with the administration of the other, or may be 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, 2 weeks, 3 weeks, or 4 weeks after the administration of the other. It is preferable that at least one of the particles or the immunoactivator is administered when it can demonstrate its activity in vivo. It is also preferable that the other is administered before the activity of the previously administered particles or immunoactivator in vivo is lost. Alternatively, the particles or the immunoactivator may be administered first.

[0068] As used herein, the term "combination drug" refers to a drug containing multiple independent preparations for use in combination as described above. The combination drug of the present invention is a drug for use in treatment and / or prevention, and contains the particles and the immune activator as independent preparations. Furthermore, the use of the combination drug of the present invention is preferably characterized in that the particles and the immune activator are administered separately using different administration methods. The treatment and / or prevention using the combination drug of the present invention is similar to the method for treating and / or preventing cancer described below.

[0069] The method of administration of the immune activator is not particularly limited as long as the object of the present invention is achieved, but preferably is a route different from that of the particles, and is preferably administered locally to a tumor. The administration route may be oral or parenteral, and specific dosage forms include injections, intranasal administrations, pulmonary administrations, and transdermal administrations. Injections can be administered systemically or locally by, for example, intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, or intratumoral injection, but are preferably administered by intratumoral injection. Examples of transdermal administrations include liniments or topical medications. Topical medications include solids, liquids, sprays, ointments, creams, and gels, but are preferably administered transdermally as creams.

[0070] The administration method used for local administration to a tumor is not particularly limited as long as the active ingredient can reach the tumor of the subject without going through the systemic circulation. For example, it is sufficient if the active ingredient can be retained within the tumor or in the peritumoral area. Specifically, administration by injection, such as intratumoral injection or subcutaneous injection into the skin surrounding the tumor, or transdermal administration, such as application of a topical agent to the surface of the skin surrounding the tumor, can be used as local administration to a tumor.

[0071] The active ingredient of the pharmaceutical of the present invention may contain, in addition to the particles and the immune activator, an antitumor agent known in the literature, etc., within the scope of not inhibiting the effects of the pharmaceutical of the present invention. There are no particular limitations on the known antitumor agent, but specific examples include paclitaxel, doxorubicin, daunorubicin, cyclophosphamide, methotrexate, 5-fluorouracil, thiotepa, busulfan, improsulfan, piposulfan, benzodopa, carboquone, meturedopa, uredopa, altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophene, methylparaben ... Triethylenethiophosphoramide, trimethylolomelamine, bullatacin, bullatacinone, camptothecin, bryostatin, calystatin, cryptophycin 1, cryptophycin 8, dolastatin, duocarmycin, eleutherobin, pancratistatin, sarcodictyin, spongistatin, chlorambucil, chlornaphadine (chloR NAphazine), colofosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard, carmustine, chlorozotocin, fotemustine (fotemustine), lomustine, nimustine, ranimustine, calicheamicin, dynemycin, clodronate, esperamicin, aclacinomycin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin,Detorbicin, 6-diazo-5-oxo-L-norleucine, adriamycin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin C, mycophenolic acid acid), nogalamycin, olivomycin, peplomycin, potfilomycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, denopterin, pteropterin, trimetrexate, fludarabine (f ludarabine), 6-mercaptopurine, thiamiprine, thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, aminoglutethimide, mitotane, trilostane, frolinic acid acid), aceglatone, aldophosphamide glycoside, aminolevulinic acid, eniluracil, amsacrine, bestrabucil, bisantrene, edatraxate, defofamine, demecolcine, diaziquone, elfornithine, elliptinium acetate, epothilone, etoglucid, lentinan, lonidamine,Maytansine, ansamitocine, mitoguazone, mitoxantrone, mopidanmol, nitraerine, pentostatin, phenamet, pirarubicin, losoxantrone, podophyllinic acid, 2-ethylhydrazide, procarbazine, razoxane, rhizoxin, sizofiran, spirogermanium, tenuazonic acid acid), triaziquone, roridin A, anguidine, urethane, vindesine, dacarbazine, mannomustine, mitobronitol, mitolactol, pipobroman, gacytosine, doxetaxel, gemcitabine, 6-thioguanine, mercaptopurine, cisplatin, oxaliplatin, carboplatin, vinbrain, Examples of anti-inflammatory drugs include benzodiazepine, etoposide, mitoxantrone, vincristine, vinorelbine, novantrone, teniposide, edatrexate, daunomycin, aminopterin, xeloda, ibandronate, irinotecan, topoisomerase inhibitors, difluoromethylolnitine (DMFO), retinoic acid, capecitabine, and pharmaceutically acceptable (known) salts or (known) derivatives thereof.

[0072] The administration method of the antitumor agent used in combination is not particularly limited. For example, it may be the same as or different from the administration method of the particles or immune activators. The administration method is oral or parenteral, and specific dosage forms include injections, intranasal administrations, pulmonary administrations, and transdermal administrations. Injections can be administered systemically or locally by, for example, intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, or intratumoral injection. Examples of transdermal administrations include liniments or topical medications. Topical medications include solids, liquids, sprays, ointments, creams, and gels.

[0073] The treatment and / or prevention of cancer according to the present invention includes various forms in addition to administration as a pharmaceutical agent as described above.

[0074] Specifically, the method for treating and / or preventing cancer of the present invention is not particularly limited as long as the object of the present invention is achieved, but preferably is characterized in that the particles (particles comprising an amphiphilic polymer, the hydrophobic segment of which is poly(hydroxy acid) and the hydrophilic segment of which is polysaccharide, and a cancer antigen) and the immune activator are administered separately by different administration methods. Preferably, the method for treating and / or preventing cancer of the present invention includes a step of administering the particles to a subject and a step of administering the immune activator to a subject, but as described above in relation to "combination use," either step may be performed first, or both steps may be performed simultaneously. Furthermore, if necessary, surgical procedures or administration of other pharmaceuticals (e.g., the known antitumor agents described above) may also be performed.

[0075] For example, the active ingredients of the combination pharmaceutical of the present invention can be administered simultaneously or separately in a sequential order. Specifically, the second active ingredient can be administered within a time interval of up to about three weeks, i.e., immediately after the administration of the first active ingredient and up to about three weeks after. For example, administration can be performed following a surgical procedure, or a surgical procedure can be performed between the administration of the first and second active ingredients. The pharmaceutical of the present invention can also be administered in multiple administration cycles. For example, when the active ingredients of the pharmaceutical of the present invention are administered simultaneously, both pharmaceuticals containing the active ingredients can be administered in a cycle lasting from about two days to about three weeks. Thereafter, the cycle can be repeated as needed, according to the judgment of the physician in charge of the treatment cycle. Even if the active ingredients are initially administered simultaneously, subsequent administrations of the pharmaceutical containing each ingredient can be administered in different administration cycles. In this case, the administration periods of the individual active ingredients other than the first one are adjusted so that they cover the same period. Similarly, when a sequential regimen is planned, the administration periods of the individual active ingredients are adjusted so that they cover the same period. The interval between cycles can vary from 0 to 2 months. The dosage of each active ingredient of the agent for treating and / or preventing cancer of the present invention can be set in the same manner as the dosage of each active ingredient in a pharmaceutical product.

[0076] The administration period of the active ingredient is not particularly limited. Specifically, it can be, for example, 3 days or more, 4 days or more, 5 days or more, 1 week or more, 10 days or more, or 2 weeks or more. Furthermore, the number of administrations of each ingredient during the administration period is also not particularly limited. As described above, it is preferable that each active ingredient is administered so that its activity is obtained over the same administration period. Therefore, the number of administrations or frequency of administration can be appropriately determined depending on the period during which each active ingredient exhibits activity in the body. Specifically, it may be determined, for example, depending on the administration method, dosage, type of active ingredient, etc. Although not particularly limited, for example, in the case of an injection, it can be administered at a frequency of once a month or more, once every 3 weeks or more, once every 20 days or more, once every 10 days or more, once a week or more, or twice a week or more. Furthermore, for example, in the case of a topical agent, it can be administered at a frequency of once every 10 days or more, once a week or more, twice a week or more, once every 3 days or more, once every 2 days or more, or once a day or more.

[0077] The cancers targeted by the present invention are not particularly limited as long as they express the cancer antigen contained in the particles. Preferred are the aforementioned basal cell carcinoma, Paget's disease, skin cancer, breast cancer, kidney cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cell tumor, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, and head and neck cancer. Palpable cancers, subcutaneous cancers, intracutaneous cancers, superficial cancers, dermal cancers, and non-parenchymal organ cancers are more preferred. Furthermore, these cancers may be primary cancers, metastatic cancers, metastatic cancers, or recurrent cancers.

[0078] More specifically, the cancers include, for example, Bowen's disease, melanoma, squamous cell carcinoma, extramammary Paget's disease, mycosis fungoides, Sezary's syndrome, cutaneous T / NK cell lymphoma, T-cell leukemia / lymphoma with lesions only in the skin, cutaneous B-cell lymphoma (indolent group), cutaneous T-cell lymphoma of the breast, combined breast adenocarcinoma, malignant mixed tumor of the breast, intraductal papillary adenocarcinoma, lung adenocarcinoma, and malignant mixed tumor of the breast. Squamous cell carcinoma, small cell carcinoma, large cell carcinoma, neuroepithelial tissue tumors such as glioma, glioblastoma, neuroblastoma, ependymoma, neuronal cell tumor, embryonal neuroectodermal tumor, schwannoma, neurofibroma, meningioma, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, digestive lymphoma, small to medium cell lymphoma, cecal cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, sigmoid colon cancer, and rectal cancer , ovarian epithelial carcinoma, germ cell tumor, stromal cell tumor, ductal carcinoma of the pancreas, invasive ductal carcinoma of the pancreas, adenocarcinoma of the pancreas, acinar cell carcinoma, adenosquamous carcinoma, giant cell tumor, intraductal papillary mucinous tumor of the pancreas, mucinous cystadenocarcinoma, pancreatoblastoma, head cell tumor of the pancreas, Frantz tumor, serous cystadenocarcinoma, solid papillary carcinoma, gastrinoma, glucagonoma, insulinoma, multiple endocrine neoplasia 1 (Wermer syndrome), non-functioning islet Cell carcinoma, somatostatinoma, VIP-producing tumor, cervical cancer, endometrial cancer, fibrosarcoma, osteo-articular sarcoma, Ewing's sarcoma, Wilms' tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, malignant soft tissue tumor, teratoma group tumor, head and neck cancer includes, but is not limited to, hypopharyngeal cancer, oropharyngeal cancer, tongue cancer, nasopharyngeal cancer, oral cancer, lip cancer, paranasal sinus cancer, laryngeal cancer, etc. Also included are palpable cancers, subcutaneous cancer, intracutaneous cancer, superficial cancer, dermal cancer, and non-parenchymal cancers that originate from the above cancers. Also included are palpable cancers, subcutaneous cancer, intracutaneous cancer, superficial cancer, dermal cancer, and non-parenchymal cancers that metastasize or recur from the above cancers.

[0079] The combination of the particles and the immune activator, which are pharmaceuticals of the present invention, has cytotoxic activity in vivo. Therefore, the antitumor effect of the present invention can be determined by examining the cytotoxic activity against cancer. The cytotoxic activity can be evaluated by administering the particles and the immune activator in combination to a living body (subject) with cancer, measuring the tumor size after administration, and examining the cancer size over time. The antitumor effect of the present invention can also be evaluated by examining the survival rate of the subject. It can also be evaluated by examining the ability to produce cytokines or chemokines. The antitumor effect of the combination of the particles and the immune activator of the present invention can also be determined by examining the prevention of cancer, metastasis, or recurrence.

[0080] Furthermore, preferred subjects (patients) are mammals, including, for example, primates, pet animals, livestock, sport animals, etc., with humans, dogs, and cats being particularly preferred.

[0081] The pharmaceuticals of the present invention can be formulated by methods known to those skilled in the art. For example, they can be used parenterally in the form of injections, such as sterile solutions or suspensions mixed with water or other pharmaceutically acceptable liquids. For example, they can be formulated by appropriately combining them with pharmacologically acceptable carriers or vehicles, specifically, sterilized water, physiological saline, vegetable oils, emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, excipients, binders, etc., and mixing them in a unit dosage form required for generally accepted pharmaceutical practice. The amount of active ingredient in these formulations is such that an appropriate dose within the indicated range can be obtained.

[0082] Sterile compositions for injection can be formulated according to standard pharmaceutical practice using a vehicle such as distilled water for injection. Examples of aqueous solutions for injection include isotonic solutions containing physiological saline, glucose, or other adjuvants, and aqueous solutions containing D-sorbitol, D-mannose, D-mannitol, or sodium chloride. These solutions may be used in combination with suitable solubilizers, such as alcohols, specifically ethanol, polyalcohols such as propylene glycol and polyethylene glycol, or nonionic surfactants such as polysorbate 80™ and HCO-60. Examples of oily solutions include sesame oil and soybean oil, which may be used in combination with benzyl benzoate or benzyl alcohol as a solubilizer. Buffers such as phosphate buffers and sodium acetate buffers, soothing agents such as procaine hydrochloride, stabilizers such as benzyl alcohol, phenol, and antioxidants may also be added. The prepared injection solutions are usually filled into appropriate ampoules.

[0083] Further aspects are described below. The present invention also relates to the use of each active ingredient in combination with other active ingredients as described above. Specifically, the present invention relates to the use of the particles (particles comprising an amphiphilic polymer, the hydrophobic segment of which is a poly(hydroxy acid) and the hydrophilic segment of which is a polysaccharide, and a cancer antigen) in the treatment and / or prevention of cancer. The use herein comprises administering the particles in combination with an immune activator to a subject, preferably wherein the administration of the particles and the administration of the immune activator are carried out separately by different administration methods.

[0084] Similarly, the present invention relates to the use of an immune activator in the treatment and / or prevention of cancer, which comprises administering the immune activator to a subject in combination with the particles, preferably characterized in that the administration of the particles and the administration of the immune activator are carried out separately by different administration methods.

[0085] The present invention further relates to a composition containing each active ingredient for use in the above-mentioned method. Specifically, the present invention relates to a composition containing the above-mentioned particles for use in the treatment and / or prevention of cancer. The use herein comprises administering the composition in combination with an immune activator to a subject, preferably characterized in that the administration of the composition and the administration of the immune activator are carried out separately by different administration methods. This use can include, for example, the steps of administering the composition to a subject and administering the immune activator to a subject.

[0086] Similarly, the present invention relates to a composition for use in the treatment and / or prevention of cancer, comprising an immune activator. The use herein comprises administering the composition to a subject in combination with the particles, preferably characterized in that the administration of the particles and the administration of the composition are carried out separately by different administration methods. This use may, for example, comprise a step of administering the particles to a subject and a step of administering the composition to a subject. Here, the administration of these compositions may be carried out in the same dosage, administration method, and administration frequency as described above for the administration of the active ingredient.

[0087] The use in these aspects is similar to the content described above in relation to the method for treating and / or preventing cancer and the use of the pharmaceutical of the present invention. For example, these uses may include a step of administering the particles to a subject and a step of administering the immune activator to a subject, and the administration of each active ingredient may be performed using the dosage, administration method, administration interval, administration time, administration number, etc. described above for each active ingredient.

[0088] Examples are shown below, but the present invention is not limited to these Examples. (Example 1) Synthesis of dextran-PLGA In this example, a linear block polymer was synthesized by adding poly(lactic acid-glycolic acid) (PLGA) to dextran and carrying out a condensation reaction.

[0089] <Dextran having a primary amino group at the reducing end (dextran-NH ２Synthesis of Dextran-NHBoc> Sodium triacetoxyborohydride (203.5 mg) and N-Boc-ethylenediamine (76.1 μl) were added to a dimethyl sulfoxide solution of dextran (number average molecular weight 2,100, PHARMACOSMOS) at a concentration of 500 mg / 2 ml, and the reaction was carried out with stirring at 60°C for 91 hours. Next, unreacted N-Boc-ethylenediamine was removed by dialysis using water as the external solution, and the mixture was freeze-dried to synthesize dextran-NHBoc. Next, the Boc terminal group of dextran-NHBoc (450 mg) was deprotected (35% aqueous hydrochloric acid solution (5 ml) / dimethyl sulfoxide (5 ml), stirred at room temperature for 35 hours), and then purified by water dialysis and freeze-dried to obtain a polymer powder (dextran-NH ２ ) was recovered.

[0090] Dextran-NH ２ The number average molecular weight of dextran-NH was determined by GPC measurement (column: TSK-gel α-5000 × 2 manufactured by Tosoh Corporation, DMF solvent, detector: RI, standard: pullulan) (Figure 1: Dextran-NH ２ ). １ H-NMR measurement confirmed that a primary amino group had been introduced into the reducing end of dextran.

[0091] <Synthesis of dextran-PLGA> Poly(lactic acid-glycolic acid) having heterobifunctional terminal groups (COOH-PLGA-OH, Fujifilm Wako Pure Chemical Industries, Ltd., PLGA-5020, number average molecular weight 8,900) (350 mg) was mixed with a solution of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (35.5 mg) and N-hydroxysuccinimide (NHS) (21.3 mg) in dimethyl sulfoxide (0.58 ml), and the mixture was reacted at 50°C for 2 hours to convert the carboxyl group at the α-terminus to an active ester (NHS). A dextran having a primary amino group at the reducing end (dextran-NH ２ , number average molecular weight 2,700) (100 mg) was added, and the active ester of NHS-PLGA-OH (NHS) and dextran-NH ２The condensation reaction with the primary amino group of the dextran-NH was carried out for 110.5 hours. After the reaction, the unreacted dextran-NH was removed by water dialysis. ２ The unreacted NHS-PLGA-OH was removed by ultracentrifugation purification to obtain dextran-PLGA polymer 1.

[0092] In addition, dextran-NH ２ Dextran-PLGA polymer 2 was obtained using PLGA-5020 (number average molecular weight 1,800) and PLGA-5020 (number average molecular weight 8,900).

[0093] The number-average molecular weight of dextran-PLGA was determined by GPC measurement (column: Toso TSK-gel α-5000 × 2, DMF solvent, detector: RI, standard: pullulan) (Table 1, Figure 1: dextran-PLGA). １ H-NMR measurement confirmed that the condensation reaction had progressed (Figure 2: dextran-PLGA).

[0094]

[0095] Example 2: Synthesis of black yeast glucan-PLGA In this example, a linear block polymer was synthesized by adding PLGA to black yeast glucan and carrying out a condensation reaction.

[0096] <Black yeast glucan having a primary amino group at the reducing end (Black yeast glucan-NH ２ 5 g of black yeast glucan (Daiso Co., Ltd.) was dissolved in 150 ml of dimethyl sulfoxide, and then 5 ml of 35% aqueous hydrochloric acid solution was added and stirred for 20 minutes at 105° C. The reaction solution was transferred to a dialysis membrane and dialyzed in water, followed by freeze-drying to obtain a black yeast glucan hydrolysate (number average molecular weight 2,400) as a powder.

[0097] Sodium triacetoxyborohydride (413.3 mg) and N-Boc-ethylenediamine (246.9 μl) were added to a 2.5 g / 8 ml dimethyl sulfoxide solution of black yeast glucan hydrolysate (number average molecular weight 2,400), and the reaction was carried out at 55°C for 169.5 hours with stirring. Next, unreacted N-Boc-ethylenediamine was removed by dialysis against water as the external solution, yielding black yeast glucan-NHBoc. The Boc terminal group of black yeast glucan-NHBoc (1.19 g) was then deprotected (35% aqueous hydrochloric acid solution (14.3 ml) / dimethyl sulfoxide (24 ml), stirred at room temperature for 2 hours), purified by water dialysis, and freeze-dried to obtain a polymer powder (black yeast glucan-NHBoc). ２ ) was recovered.

[0098] Black yeast glucan-NH ２ The number average molecular weight of the black yeast glucan-NH was determined by GPC measurement (column: TSK-gel α-5000 × 2 manufactured by Tosoh Corporation, DMF solvent, detector: RI, standard: pullulan) (Figure 3: Black yeast glucan-NH ２ ). １ H-NMR measurement confirmed that a primary amino group had been introduced into the reducing end of the black yeast glucan.

[0099] <Synthesis of Black Yeast Glucan-PLGA> Poly(lactic acid-glycolic acid) having heterobifunctional terminal groups (COOH-PLGA-OH, Fujifilm Wako Pure Chemical Industries, Ltd., PLGA-5020, number average molecular weight 8,900) (4.462 g) was mixed with a DMSO (2.5 ml) solution of EDC (479.25 mg) and NHS (287.73 mg), and the mixture was allowed to react at 50°C for approximately 18 hours to convert the α-terminal carboxyl group into an active ester (NHS). Black yeast glucan having a primary amino group at the ω-terminal (Black Yeast Glucan-NH ２ , number average molecular weight 1,500) (749 mg) was added, and the active ester of NHS-PLGA-OH (NHS) and black yeast glucan-NH ２ After the reaction, unreacted black yeast glucan-NH was removed by water dialysis. ２After removing the residue, unreacted NHS-PLGA-OH was removed by ultracentrifugation purification to obtain black yeast glucan-PLGA polymer 3.

[0100] In addition, black yeast glucan-NH was synthesized in the same manner as in black yeast glucan-PLGA polymer 3. ２ (number average molecular weight 1,200) and PLGA-5020 (number average molecular weight 8,900) were used to obtain black yeast glucan-PLGA polymer 4, and black yeast glucan-NH ２ (number average molecular weight 2,300) and PLGA-5020 (number average molecular weight 8,900) were used to obtain black yeast glucan-PLGA polymer 5.

[0101] The number-average molecular weight of black yeast glucan-PLGA was determined by GPC measurement (column: TSK-gel α-5000 x 2 manufactured by Tosoh Corporation, DMF solvent, detector: RI, standard: pullulan) (Table 2, Figure 3: black yeast glucan-PLGA). １ H-NMR measurement confirmed that the condensation reaction had progressed (Figure 4: Black yeast glucan-PLGA).

[0102]

[0103] Example 3: Preparation of particles using the S / O / W emulsion method (OVA-containing dextran particles (1), OVA-containing black yeast glucan particles (2)). A polymer solution was prepared by dissolving 30 mg of dextran-PLGA polymer powder (Table 1, amphiphilic polymer 1) in a mixture of 1.2 ml of dimethyl carbonate and 133 μl of tert-butanol. 0.3 ml of a 0.5% (w / v) aqueous solution of OVA (ovalbumin, Sigma) was added dropwise to the polymer solution, and a W / O emulsion solution was produced by stirring for 1 minute at 11,000 rpm using a mixer (Polytron, PT2100S). The W / O emulsion solution was pre-frozen with liquid nitrogen and then freeze-dried for 4 hours using a freeze dryer (Tokyo Rikakikai Co., Ltd., FD-1000) at a trap cooling temperature of −45°C and a vacuum of 20 Pa. The resulting solid was dispersed in 3 ml of ethyl acetate to prepare an S / O suspension. The S / O suspension was added dropwise to 12 ml of 1% (w / v) aqueous polyvinyl alcohol solution and stirred for 3 minutes at 5,000 rpm using a Silverson L5M-A mixer to prepare an S / O / W emulsion. The ethyl acetate was removed from the S / O / W emulsion by submerged drying to yield a particle suspension. The suspension was transferred to a 50 ml tube and centrifuged at 3,000 g for 30 minutes to precipitate the particles. After removing the supernatant, the particles were washed by resuspending them in 50 ml of distilled water and centrifuging under the same conditions as above to precipitate the particles again. This washing procedure was repeated once more, and the supernatant was removed. The resulting particles were then suspended in 2.4 ml of an aqueous solution containing 5% (w / v) mannitol and 0.1% (w / v) polysorbate 80. The suspension was filtered through a mesh filter (1 μm), pre-frozen with liquid nitrogen, and then freeze-dried using a freeze dryer at a trap cooling temperature of −45° C. and a vacuum of 20 Pa for 12 hours to obtain OVA-containing dextran particles (1).

[0104] In addition, OVA-containing black yeast glucan particles (2) were obtained using a black yeast glucan-PLGA polymer (Table 2, amphiphilic polymer 3) in the same manner as in the above-mentioned OVA-containing dextran particles.

[0105] The evaluation results for each particle are shown in Table 3. The average particle size of the particles was calculated by the cumulant method using a dynamic light scattering device (ELS-Z, Otsuka Electronics Co., Ltd.) (Table 3, Figure 5: OVA-containing dextran particles, Figure 6: OVA-containing black yeast glucan particles). The OVA antigen content (w / w) was determined by extracting the antigen from the particles using an organic solvent, subjecting the extracted antigen to gel electrophoresis using a gel electrophoresis device (TEFCO), and then staining using a colloidal CBB staining kit (TEFCO).

[0106]

[0107] (Example 4) Antitumor effect of combined use of OVA-containing dextran particles (1) and imiquimod The OVA-containing dextran particles (1) prepared in Example 3 and imiquimod were administered separately, and the antitumor effect in vivo in tumor-bearing mice was evaluated.

[0108] Specifically, the antitumor effect of the combined use of OVA-containing dextran particles (1) and imiquimod was investigated using C57BL / 6NCR mice subcutaneously implanted with OVA-expressing cancer cells. ６ Each mouse lymphoma cell line, EG7, was subcutaneously transplanted, and starting on the seventh day after transplantation, 0.5 mg of the OVA-containing dextran particles (1) prepared in Example 3 (excluding mannitol) was suspended in PBS and water for injection to achieve isotonicity and administered subcutaneously to the right armpit of each mouse twice a week, once a week. Imiquimod application was initiated simultaneously with the first administration of the OVA-containing dextran particles (1) to the same mice. Veselna Cream 5% (Mochida Pharmaceutical Co., Ltd., hereafter referred to as "Imiquimod Cream"), containing imiquimod as an active ingredient, was applied to the surface of the skin where the cancer cells had been transplanted, for five consecutive days per week. After two days without application, imiquimod cream was applied for five consecutive days starting on the eighth day after the start of administration, as described above.

[0109] As a comparative control group, only the same OVA-containing dextran particles (1) as above were administered to tumor-bearing mice at the same dose and administration interval. Furthermore, as a comparative control group, only imiquimod cream was applied to the surface of the skin where cancer cells had been transplanted at the same administration interval to other tumor-bearing mice. Furthermore, tumor-bearing mice in the untreated group served as a negative control. After the start of administration, the size of the tumor in the tumor-bearing mice was measured over time using a vernier caliper. Tumor volume was calculated according to the standard method using the formula: (length of the longest axis of the tumor) x (length of the shortest axis of the tumor). ２ The calculation was made by multiplying the ratio by 0.5.

[0110] As a result of the evaluation, the tumors of the mice administered with the OVA-containing dextran particles (1) prepared in Example 3 and the application of imiquimod cream regressed compared to the start of administration. In contrast, the tumors of the mice administered with only the OVA-containing dextran particles (1) prepared in Example 3 showed delayed growth compared to the negative control, but no regression was observed, and the tumors of the cancer-bearing mice administered with only imiquimod cream showed no change compared to the negative control.

[0111] The results of this evaluation showed that administering OVA-containing dextran particles and imiquimod in combination separately had a significant antitumor effect compared to administering OVA-containing dextran particles alone or administering imiquimod alone.

[0112] (Example 5) Antitumor effect of combined use of OVA-containing black yeast glucan particles (2) and imiquimod OVA-containing black yeast glucan particles (2) prepared in Example 3 and imiquimod were administered separately, and the antitumor effect in vivo in tumor-bearing mice was evaluated.

[0113] Specifically, the antitumor effect of the combined use of OVA-containing black yeast glucan particles (2) and imiquimod was investigated using C57BL / 6NCR mice subcutaneously implanted with OVA-expressing cancer cells. ６Each mouse lymphoma cell line, EG7, was subcutaneously transplanted, and starting on the seventh day after transplantation, 0.5 mg of the OVA-containing black yeast glucan particles (2) prepared in Example 3 (excluding mannitol) was suspended in PBS and water for injection to an isotonicity and administered subcutaneously to the right armpit of each mouse twice a week, once a week. Imiquimod application was initiated simultaneously with the first administration of the OVA-containing black yeast glucan particles (2). Imiquimod cream was applied to the surface of the skin where the cancer cells had been transplanted for five consecutive days per week. No administration was performed for the next two days, and then, starting on the eighth day after the start of administration, imiquimod cream was applied for five consecutive days in the same manner as above.

[0114] As a comparative control group, only the same OVA-containing black yeast glucan particles (2) as above were administered to tumor-bearing mice at the same dose and administration interval. Furthermore, as a comparative control group, only imiquimod cream was applied to the surface of the skin where cancer cells had been transplanted at the same administration interval to other tumor-bearing mice. Furthermore, tumor-bearing mice in the untreated group served as a negative control. After the start of administration, the size of the tumor in the tumor-bearing mice was measured over time using a vernier caliper. Tumor volume was calculated according to the standard method using the formula: (length of the longest axis of the tumor) x (length of the shortest axis of the tumor). ２ The calculation was made by multiplying the ratio by 0.5.

[0115] As a result of the evaluation, the tumors of the mice administered with the OVA-containing black yeast glucan particles (2) prepared in Example 3 and the application of imiquimod cream regressed compared to the start of administration. In contrast, the tumors of the mice administered with only the OVA-containing black yeast glucan particles (2) prepared in Example 3 showed delayed growth compared to the negative control, but no regression was observed, and the tumors of the cancer-bearing mice administered with only imiquimod cream showed no change compared to the negative control.

[0116] The results of this evaluation showed that administering OVA-containing black yeast glucan particles and imiquimod in combination separately had a significant antitumor effect compared to administering OVA-containing black yeast glucan particles alone or administering imiquimod alone.

[0117] (Example 6) Antitumor effect of combined use of OVA-containing black yeast glucan particles (2) and Poly(I:C) The OVA-containing black yeast glucan particles (2) prepared in Example 3 and Poly(I:C) were administered separately, and the antitumor effect in vivo in tumor-bearing mice was evaluated.

[0118] Specifically, the antitumor effect of the combined use of OVA-containing black yeast glucan particles and Poly(I:C) (Sigma, catalog number: P1530) was investigated using C57BL / 6NCR mice subcutaneously implanted with OVA-expressing cancer cells. ６ Each mouse lymphoma cell line, EG7, was subcutaneously transplanted, and starting on the seventh day after transplantation, 0.5 mg of the OVA-containing black yeast glucan particles (2) prepared in Example 3 (excluding mannitol) was suspended in a solvent of PBS and water for injection to an isotonicity and administered subcutaneously to the right armpit of each mouse, once a week for a total of two doses. Poly(I:C) was administered intratumorally to the same mice simultaneously with the initial administration of the OVA-containing black yeast glucan particles (2). Poly(I:C) was administered at a dose of 50 μg twice a week for a total of four doses.

[0119] As a comparative control group, only the same OVA-containing black yeast glucan particles (2) as above were administered to tumor-bearing mice at the same dose and administration interval. Furthermore, as a comparative control group, only Poly(I:C) was administered intratumorally to another tumor-bearing mouse at the same dose and administration interval. Furthermore, a mixture of OVA-containing black yeast glucan particles (2) and Poly(I:C) was administered subcutaneously to the right armpit of another tumor-bearing mouse at the same dose and administration interval. Untreated tumor-bearing mice served as negative controls. After the start of administration, the size of the tumor in the tumor-bearing mice was measured over time using calipers. Tumor volume was calculated according to the standard method using the formula: (length of the longest axis of the tumor) x (length of the shortest axis of the tumor). ２ The calculation was made by multiplying the ratio by 0.5.

[0120] As a result of the evaluation, the tumors of mice that had been administered intratumorally with OVA-containing black yeast glucan particles (2) prepared in Example 3 and Poly(I:C) separately showed regression compared to the start of administration. In contrast, the tumors of mice that had been administered only with OVA-containing black yeast glucan particles (2) prepared in Example 3 showed delayed growth compared to the negative control, but no regression was observed, and the tumors of the cancer-bearing mice that had been administered only with Poly(I:C) showed no change compared to the negative control. Furthermore, the tumors of mice that had been administered a mixture of OVA-containing black yeast glucan particles (2) and Poly(I:C) showed delayed growth, but no regression was observed.

[0121] The results of this evaluation showed that administering OVA-containing black yeast glucan particles and Poly(I:C) in combination separately had a significant antitumor effect compared to administering OVA-containing black yeast glucan particles alone, administering Poly(I:C) alone, or administering a mixture of OVA-containing black yeast glucan particles and Poly(I:C).

[0122] The combination drug of the present invention for use in the treatment and / or prevention of cancer, which comprises particles containing an amphiphilic polymer in which the hydrophobic segment is a poly(hydroxy acid) and the hydrophilic segment is a polysaccharide, a cancer antigen, and an immune activator, and is characterized in that the particles and the immune activator are administered separately, can be used particularly for the treatment and / or prevention of infectious diseases, cancer, etc.

[0123] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A combination pharmaceutical for the treatment and / or prevention of cancer, comprising a particle containing an amphiphilic polymer in which a hydrophobic segment is a poly(hydroxy acid) and a hydrophilic segment is a polysaccharide and a cancer antigen, and an immunostimulatory factor, wherein the use is characterized in that the administration of the particle and the administration of the immunostimulatory factor are performed separately.

2. The pharmaceutical according to claim 1, wherein the use is characterized in that the administration of the particle and the administration of the immunostimulatory factor are performed separately by different administration methods.

3. The pharmaceutical according to claim 1 or 2, wherein the immunostimulatory factor is a ligand or agonist that binds to a Toll-like receptor (TLR), NOD-like receptor (NLR), RIG-like receptor, C-type lectin receptor (CLR) or stimulator of interferon genes (STING).

4. The pharmaceutical according to claim 3, wherein the ligand or agonist that binds to the Toll-like receptor (TLR) is a ligand or agonist that binds to TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9 or TLR11.

5. The pharmaceutical according to claim 4, wherein the ligand or agonist that binds to the Toll-like receptor (TLR) is any one of the following (i) to (vii). (i) A ligand or agonist that binds to TLR2 selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide and zymosan (ii) A ligand or agonist that binds to TLR3 selected from the group consisting of Poly(I:C) and poly(A:U) (iii) A ligand or agonist that binds to TLR4 selected from the group consisting of lipopolysaccharide (LPS), HSP60, RS09 and MPLA (iv) Flagellin, which is a ligand or agonist that binds to TLR5 (v) A ligand or agonist that binds to TLR7 or 8 selected from the group consisting of imidazoquinoline compounds and single-stranded RNA (vi) A ligand or agonist that binds to TLR9 selected from the group consisting of bacterial DNA, unmethylated CpG DNA, hemozoin, ODN1585, ODN1668 and ODN1826 (vii) A ligand or agonist that binds to TLR11 selected from the group consisting of profilin and uropathogenic bacteria

6. The pharmaceutical product according to claim 5, wherein the imidazoquinoline compound is imiquimod.

7. The pharmaceutical product according to claim 1 or 2, wherein the administration of the immunostimulatory factor is administration to the tumor site.

8. The pharmaceutical product according to claim 1 or 2, wherein the polysaccharide is dextran, β-glucan, mannan, chitin, chitosan, gellan gum, alginic acid, hyaluronic acid or pullulan.

9. The pharmaceutical product according to claim 8, wherein the β-glucan is a polymer of glucose linked by one or more β-1,3 linkages and / or one or more β-1,6 linkages.

10. The pharmaceutical product according to claim 8, wherein the β-glucan is Aureobasidium pullulans glucan, curdlan, pachyman, laminaran, lichenan, schizophyllan, lentinan, scleroglucan or pachymaran.

11. The pharmaceutical product according to claim 1 or 2, wherein the poly(hydroxy acid) is poly(lactic-co-glycolic acid), polylactic acid or polyglycolic acid.

12. The pharmaceutical product according to claim 1 or 2, wherein the number average molecular weight of the amphiphilic polymer is 500 to 100,000.

13. The pharmaceutical product according to claim 1 or 2, wherein the average particle size of the particles is 0.1 to 50 μm.

14. The pharmaceutical product according to claim 1 or 2, wherein the cancer is basal cell carcinoma, Bowen's disease, skin cancer, breast cancer, renal cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or head and neck cancer.