JP2009528267A - Glucan to enhance treatment - Google Patents

Abstract

  Disclosed are therapeutic compositions for treating cancer in mammals. The composition comprises an effective amount of a yeast β-glucan composition suitable for oral administration and absorption by the mammalian gastrointestinal tract. The therapeutic composition may further comprise an anti-tumor antibody or cancer vaccine composition, wherein the anti-tumor activity of the anti-tumor antibody or cancer vaccine composition is enhanced by yeast glucan.

Description

  This application is a continuation-in-part of US patent application Ser. No. 11 / 334,763 filed Jan. 17, 2006, which is hereby incorporated by reference.

  Throughout this application, various references are cited. The disclosures of these publications are hereby incorporated by reference in their entirety in order to more fully describe the state of the art to which this invention pertains.

  Glucans are a heterogeneous group of glucose polymers found in the cell walls of plants, bacteria, and fungi. The basic structure of a branched glucan consists of a β-1,3-glucan side chain having a β-1,6-linked, β-1,3-linked glucose linear backbone. β-1,3-glucan is a component responsible for most of the biological activity of zymosan, a commonly used leukocyte stimulant derived from the cell wall of baker's yeast (Saccharomyces cerevisiae) .

β-glucan has been tested for tumor therapy in mice for nearly 40 years ^{1, 2} . In Japan, several forms of mushroom-derived β-glucan such as PSK (from Cororilus vesicor), lentinan, schizophyllan, etc. are used clinically for the treatment of cancer. In a randomized trial in Japan, PSK has moderate survival in several cancer trials after gastrectomy ³ , ⁴ to remove the primary tumor, rectal surgery ⁵ , ⁶ and esophagectomy ⁷ It is raised to the extent. In breast cancer ⁸ , ⁹ and leukemia ¹⁰ , the results were less promising. Schizophyllan also moderately increases the survival rate of patients with operable gastric cancer ¹¹ , inoperable gastric cancer ¹² , ¹³ , and cervical cancer ¹⁴ . Although β-glucans are not widely used by western oncologists, botanicals containing β-glucans such as Reishi and Maitake ¹⁵ are widely used in US cancer patients as an alternative / supplemental cancer therapy. It is used.

In Europe and the United States, especially β- glucan derived from baker's yeast, as a feed additive for animals, long used as an active ingredient of human dietary supplement as ^{17, 18} in the treatment of wounds ^and skin cream formulations It has been. The basic structural unit of β-glucan is a β (1 → 3) linked glucosyl unit. Depending on the source and separation method, β-glucans have varying degrees of branching and binding in the side chain. The frequency of the side chains and the hinge structure determine its immunomodulatory effect. Β-glucans of fungal and yeast origin are usually insoluble in water but are solubilized by acid hydrolysis or by derivatization that introduces charged groups such as phosphoric acid, sulfuric acid, amine, carboxymethyl, etc. into the molecule Can be ^19-20 .

  Previous studies exploring the therapeutic effects of β-glucan did not incorporate co-administration of therapeutic monoclonal antibodies (MoAb) as part of the study design. When β-glucan is administered without co-administration of MoAb, its tumor cytotoxic effect requires the presence of naturally occurring anti-tumor antibodies, which are anti-tumor antibodies between patients and even in experimental mice. Can vary considerably. Therefore, it is interesting to determine the therapeutic effect of co-administration of β-glucan and therapeutic monoclonal antibody. It is also unclear whether a therapeutic effect can be obtained with orally administered β-glucan.

  The present invention provides a composition comprising an effective amount of yeast β-glucan capable of enhancing an immune response in a subject or host. In one embodiment, the enhanced immune response is an anti-tumor immune response, which can be induced or initiated by various techniques or procedures generally known in the art. For example, an anti-tumor immune response can be elicited by an anti-tumor antibody, a cancer vaccine, or other composition intended to elicit an immune response against cancer.

  In one embodiment, the present invention provides yeast β-glucan compositions and / or derivatives thereof that can enhance the efficacy of anti-tumor antibodies. In another embodiment, the present invention provides a β-glucan composition of yeast that can enhance the effectiveness of cancer vaccines or other compositions intended to elicit an immune response against cancer. Offer things.

  As used herein, antibody refers to any portion of an immunoglobulin molecule (eg, a monoclonal antibody) that has a specific cancer cell binding affinity by which it can exert antitumor activity. Examples are antigen binding fragments or antibody derivatives. Furthermore, the antibody used in the present invention may be a single monoclonal antibody or a combination of antibodies. The antibody may be directed against at least one epitope or multiple epitopes of the antigen, or multiple antigens. Accordingly, the present invention includes at least one antibody.

  Cancers recognized by antibodies include, but are not limited to, neuroblastoma, melanoma, non-Hodgkin lymphoma, Epstein-Barr related lymphoma, Hodgkin lymphoma, retinoblastoma, small cell lung cancer, brain tumor, leukemia, epidermoid carcinoma Prostate cancer, renal cell cancer, transitional cell carcinoma, breast cancer, ovarian cancer, lung cancer rectal cancer, liver cancer, gastric cancer, and other gastrointestinal cancers.

  It will be appreciated that various embodiments of the present invention relating to particular methods of treating tumor and cancer disease states may be relevant within the context for the treatment of numerous other tumors and / or cancers not specifically mentioned herein. Those skilled in the art will understand. Accordingly, the embodiments described herein for the particular cancer referred to should not be construed as not applicable to other cancers.

  The present invention further provides the above composition and a pharmaceutically acceptable carrier, the formation of a pharmaceutical composition therewith.

  The present invention also provides a method for treating a subject having cancer comprising administering to the subject a composition as described above.

  In one embodiment, the present invention provides a method for enhancing the tumoricidal activity of an anti-tumor antibody in a subject with cancer. In general, examples of cancer include, but are not limited to, neuroblastoma, melanoma, non-Hodgkin lymphoma, Epstein-Barr related lymphoma, Hodgkin lymphoma, retinoblastoma, small cell lung cancer, brain tumor, leukemia, epidermoid carcinoma Prostate cancer, renal cell cancer, transitional cell carcinoma, breast cancer, ovarian cancer, lung cancer, rectal cancer, liver cancer, gastric cancer, or other gastrointestinal cancer.

  The method comprises the steps of: (a) administering to the subject an anti-tumor antibody that binds to cancer cells in the subject; and (b) orally administering to the subject a composition comprising an effective amount of yeast glucan. Orally administered glucan enhances the tumoricidal activity of the antibody. In general, the antibody is a monoclonal antibody. More preferably, the antibody can activate complement or can activate an antibody-dependent cell-mediated disorder. Representative examples of anti-tumor antibodies include, but are not limited to, antibodies that recognize CD20, CD22, HER-2 / neu, CD25, epidermal growth factor receptor, or ganglioside.

  In another embodiment, the average molecular weight of the yeast glucan used in the above method is from about 16000 to about 17000 daltons, and the range is from about 6000 to about 30000 daltons, or from about 6000 to about 25000 daltons . Yeast glucan can be administered concurrently with the administration of the anti-tumor antibody or at different times. Preferably, yeast glucan is capable of priming or inducing secretion of cytokines, chemokines, or growth factors by leukocytes.

  The present invention also provides a therapeutic regimen for a subject with cancer, the regimen comprising (i) an anti-tumor antibody that binds to cancer cells in the subject, or a cancer vaccine intended to elicit an anti-tumor immune response A composition, and (ii) a synergistic anticancer effect resulting from administering an effective amount of yeast glucan orally administered to a subject to a subject in need thereof, wherein the orally administered glucan is an antibody Alternatively, it enhances the tumoricidal activity of the cancer vaccine composition. The yeast glucan can be administered at the same time as the anti-tumor antibody or cancer vaccine, or at different times. Examples of anti-tumor antibodies and yeast glucans are described above. In general, examples of cancer vaccines include whole tumor cells, lysates of tumor cells, either clinically synthesized or recombinant products that act as tumor anti-genes, Tumor cell-derived RNA, tumor cell-derived proteins, tumor cell-derived peptides, tumor cell-derived carbohydrates, tumor cell-derived lipids, tumor cell-derived DNA sequences, and genetically modified tumor cells. These tumor cells may be derived from the patient's own tumor or may be derived from a tumor from an unrelated donor.

  The present invention also provides the use of an effective amount of yeast glucan for the preparation of an anti-cancer pharmaceutical composition. Glucan is administered orally to the subject to enhance tumoricidal activity in the subject. The anti-cancer drug composition can further comprise an anti-tumor antibody that binds to cancer cells in the subject, or a cancer vaccine composition intended to elicit an anti-tumor immune response. The yeast glucan can be administered to the subject simultaneously with the administration of the anti-tumor antibody or cancer vaccine, or at different times. Representative examples of anti-tumor antibodies, cancer vaccines, and yeast glucans are described above.

  The following terms are used to describe the present invention. In the absence of a specific definition set forth herein, the terms used to describe the invention will have their ordinary meaning as understood by one of ordinary skill in the art.

  In the present invention, the meaning of the expression “higher conformation” is to define the structure of the molecule by the transformation of a group of free atoms, which is a random spatial conformation. This is the transformation of these same atoms into a more stable non-random pattern, as seen in free atoms. In other words, these atoms bond to each other and become a new molecule, a new totality. The bond in this case is chemical, i.e. by chemical bonds such as hydrogen bonds. Some of these molecules react sequentially with each other and can become other molecules of either the same size, or perhaps a larger size. Both become new wholeness again. The larger the molecule, the higher the overallity.

  In the present invention, the expression “immunostimulatory” means stimulating the immune system by inducing or increasing the activity of any component of the immune system.

  In the present invention, the expression “immunopotentiating” means the activity of one substance that enhances or increases the immunostimulatory effect of another substance.

  The ability of β-glucan to have immunopotentiating activity may be the result of the ability to present multiple epitopes for interaction with receptors on target cells, thereby causing β-glucan receptors To mimic the challenge of pathogenic organisms. Multiple interactions with such specific receptors on cells are thought to depend in part on the ability of glucans to form “higher order” structures that present multiple binding epitopes in close proximity. Yes. Soluble β-glucan formulations that possess durable interchain associations, represented by high viscosity profiles, may thus be candidates for exhibiting “immunopotentiating” capabilities.

  The term “cancer” refers to a pathological process that results in the formation and growth of cancerous or malignant neoplasms.

  The term “effective amount” is used to describe the amount of a compound that, when administered to an animal or human, results in inhibition or eradication of tumor growth or cancer spread, or other desirable immune response.

  The term “animal” is used to describe an animal, preferably a mammal, more preferably a human, for which the treatment or method according to the invention is provided.

  As used herein, the term “pharmaceutically acceptable carrier, additive, or excipient” can be combined with a suitable glucan or derivative, after which the animal, preferably a mammal, most Preferably it means a chemical composition that can be used to administer a glucan suitable for treating a human.

  Previously, in international applications PCT / US02 / 01276 and PCT / US04 / 23099, high molecular weight and high viscosity profile β-1,3 and 1,4-glucan isolated from barley were administered orally. Or in eradication or suppression of tumor cells, i. v. It has been shown to be effective in enhancing the effectiveness of the administered antibody. Compositions of soluble β-glucan in yeast with β-1,3-linked, and specific types of side chains, thereby have a higher order conformation with a higher viscosity profile than previously used Surprisingly, it has also been shown to be equally active as β-glucan from barley (see FIG. 1). The full disclosures of PCT / US02 / 01276 and PCT / US04 / 23099 are hereby incorporated by reference into this application.

  It is generally accepted that β-glucans of microbial origin, such as yeast, are recognized by specific pattern recognition receptors on immune cells as a result of phylogenetic adaptation to the detection of possible pathogens. β-glucan, for example, in the cell walls of fungi, ensures the strength and integrity of cells, which are the main structural elements, and is therefore very important for organisms. β-1,3-glucans are present in almost all fungal cells, which are highly conserved structures, the latter being essential for the so-called pathogen-associated molecular pattern (PAMP) recognized by the immune system It is. Immunologically active β-glucan can bind to a β-glucan receptor known as Dectin-1 when introduced into an organism by the digestive tract.

  Purified β-1,3-glucan having structural elements and conformations that mimic its fungal origin as recognized by immune cells, therefore, activates immunity, particularly when administered orally It is considered preferable for the realization of. β-1,3, and 1,4 glucans are not derived from microorganisms, but may interact with immune cells based on their similarity to the conserved structures on pathogenic organisms. is there. Examples of useful β-glucans include, but are not limited to, particulate and soluble yeast cell wall glucans described in PCT / IB95 / 00265 and EP0759089. Other β-1,3-glucan compositions having the ability to form interchain associations also have a high viscosity profile as described for barley β-1,3-1,4-glucan preparations. As illustrated, it is a good candidate. For example, certain preparations of lentinan, scleroglucan, and schizophyllan that exhibit durable interchain interactions may be effective. Similarly, β-1,3-glucan preparations solubilized by derivatization, such as glucan phosphate, glucan sulfate, carboxymethyl-glucan, and the retention of the immune enhancing activity of natural molecules and interchain associations, Possible active product.

  Β-glucan preparations that do not display pathogenic features can be obtained as described in Herlyn et al. v. When administered directly during systemic distribution, such as when administered ("Monoclonal antibody-dependent cytotoxicity-mediated cytotoxicity humans are stimulated by lentinan." Tumors is stimulated by lentinan.) "Jpn. J. Cancer Res., 76, 37-42 (1985)), or as described in U.S. Application No. 60/261911. p. When administered, it can again be a powerful adjuvant for immunotherapy.

  In one embodiment, the invention provides (i) a viscous and immunopotentiating yeast β-glucan composition comprising a β-1,3-binding backbone as shown in FIG. 4, and (ii) an anti-tumor antibody. A composition for achieving a cooperative therapeutic effect in an animal in need thereof, preferably a mammal, most preferably a human, is disclosed, wherein the cooperative therapeutic effect is eradication or suppression of cancer or tumor cells It is. The ability of yeast β-glucan to have immunopotentiating activity is due to its ability to present multiple epitopes to interact with receptors on target cells, thereby consolidating β-glucan receptors. , May mimic the challenge of pathogenic organisms. Such multiple interactions with specific receptors on cells are due in part to the ability of β-glucans to form “higher order” structures that are in close proximity to present multiple binding epitopes. It is considered to be a thing. Soluble β-glucan formulations that possess tolerable interchain associations may also be candidates for exhibiting “immunopotentiating” ability as represented by a high viscosity profile.

  In one embodiment, the β-glucan composition comprises yeast β-glucan having a β-1,3-linked backbone with a molecular weight (MW)> 6000 Da, attached to them and having strong interchain association. It has a side chain that results in some soluble product. Representative examples include, but are not limited to, β-1,3-glucan isolated from yeast in which β-1,3-linked side chains are tethered to the main chain by β-1,6-linkages. (See FIG. 5). Suitable forms of yeast glucan include, but are not limited to, particulate, soluble, or gel forms.

  An example of a highly active composition of yeast β-glucan is a molecular weight that interacts to give higher order conformations that exhibit immunostimulatory activity when administered orally to induce a synergistic effect with an antibody. (MW) is a mixture of soluble β-glucan chains> 6000 daltons. In one embodiment, as shown in FIG. 5, a mixture of soluble β-glucan has a linear β-1,3-glucan chain with a MW> 6000 Da, or more preferably a MW range of 6000-30000 Da. Together with branched high molecular weight β-1,3-glucan (MW> 15000 Da) with β-1,3 linked side chains extending from within the main chain.

  An example of the glucan described above is SBG (soluble beta glucan) produced by Biotec Pharmacon ASA (Tromso, Norway). SBG, a pure natural product derived from baker's yeast, has been analyzed by NMR and chemical analysis for β-1,3-linked D containing the β-1,3 and β-1,6-linked D-glucose side chains. -Non-derivatized water-soluble β-1,3 / 1,6-glucan characterized as consisting of a polymer of glucose (see FIG. 2). Preferably, the average molecular weight of the yeast glucan is from about 16000 to about 17000 daltons, and the range is from about 6000 to about 30000 daltons, or from about 6000 to about 25000 daltons.

  As shown in FIG. 2, SBG has a high molecular weight having a β-1,3-linked side chain attached to a repeating β-1,3-linked main chain by a β-1,6-linked branch point. Figure 2 shows a complex β-glucan composition having a chain and a linear β-1,3-glucan chain of medium molecular weight in the range of 6-15 kDa. SBG (soluble beta glucan) exhibits a durable interchain association, as shown by its high viscosity profile and gelation behavior (see FIG. 3). SBG is used to activate human leukocytes in vitro, for example to prime and induce cytokine production (Engstad et al. (2002), “Soluble β-1,3-glucans and lipopolysaccharides. Of the production of cytokines and blood coagulation activation in whole blood (The effect of soluble beta-1,3-glucan and lipopolysaccharide on cytokine production and coagulation activation in wol. 15). -1597), and also p. o. To modulate immune function when administered (Breivik et al., (2005), “soluble β-1,3 / 1,6-glucan from yeast inhibits experimental periodontal disease in Wistar rats (Soluble). (beta-1,3 / 1,6-glucan from most yeasts, experiential periodical dissemination in Wistar rats) ”, J. Clinical Periodontology, Vol. 32, pages 347-3) It is shown. It is preferred that the yeast glucan of the present invention has such functional characteristics to prime and induce cytokine production by human leukocytes.

  Other structural and / or structural conformations in the composition of β-1,3-glucan as described above can be readily identified or isolated by those skilled in the art according to the teachings of the present invention. P. o. It is expected to have similar therapeutic effects when administered by various routes other than. Thus, while the above is a guideline for achieving a highly powerful product, it is not a limitation on even more powerful products. Isolated structural elements of complex mixtures as described above are expected to have improved efficacy over this formulation when administered orally.

  Products with desirable structural features that give higher conformations, such as SBG, that facilitate the required interaction with reactive cells in the intestine are preferred products when administered orally. is there. Their action as immunopotentiators in synergy with anti-cancer antibodies has been shown, for example, i. p. , S. c. I. m. Or i. v. May be at least potent when administered. The functional dosage range of glucan can be easily determined by those skilled in the art. For example, when administered orally, the functional dose range is 1-500 mg / kgb. w. (Body weight) / day, more preferably 10-200 mg / kg b. w. / Day, or most preferably in the region of 20-80 mg / kg / day. In another embodiment, when administered parenterally, the functional dosage range is 0.1-10 mg / kg b. w. / Day.

  Typically, the dosage of a compound of the invention administered to an animal, preferably a human, is not limited to the type of animal, as well as the type of cancer and disease state being treated, the age of the animal, the route of administration, and It will vary depending on any number of factors, including the relative therapeutic index.

  The route of administration will be readily apparent to those skilled in the art and will depend on any number of factors including the type and severity of the disease being treated, the type and age of the human patient being treated, and the like.

  Formulations suitable for oral administration of β-glucan include, but are not limited to, aqueous or oily suspensions, aqueous or oily solutions, or emulsions. Such formulations can be administered by any means including, but not limited to, soft gelatin capsules.

  Liquid formulations of the pharmaceutical composition of the present invention suitable for oral administration are prepared either in liquid form or in the form of a dry product intended to be reconstituted with water or other suitable vehicle prior to use. Can be packaged and sold.

  In general, β-glucan can be administered to animals as often as several times a day, or may be administered less frequently, such as once a day. Antibody treatment depends on, for example, the type of antibody, the type of cancer, the severity of the cancer, and the condition of each patient. β-glucan treatment correlates closely with the antibody treatment regimen and may be before, contemporaneously with, or after antibody administration. The frequency of β-glucan and the dosage of the antibody will be readily apparent to those skilled in the art and include, but are not limited to, any number of factors such as, but not limited to, the type and severity of the disease being treated, and the type and age of the patient Depends on. Treatment with the substances of the invention may occur simultaneously or at different times. For example, β-glucan treatment is i. v. May begin several days prior to antibody treatment, and then β-glucan is administered simultaneously with the antibody. In another embodiment, β-glucan treatment may be continued for several days after completion of antibody treatment. Antibody treatment can include a cocktail of antibodies or antibody formulations, modified antibodies and / or derivatives thereof.

  When glucan is administered orally, it is taken up by macrophages and monocytes, which transports this hydrocarbon to the bone marrow and reticuloendothelial system, from which it is properly processed on bone marrow cells containing neutrophils, and natural Released on lymphocytes including killer (NK) cells. Processed glucans bind to CR3 on these neutrophils and NK cells and activate their anti-tumor cytotoxicity in the presence of tumor-specific antibodies.

  The present invention provides compositions comprising an effective amount of orally administered (1 → 3), (1 → 6) β-glucan that can enhance the efficacy of anti-tumor antibodies. Glucans derived from yeast cell walls such as Saccharomyces cerevisiae can be used in the compositions described above. Preferably, a glucan having β (1-3) and β (1-6) linkages, such as SBG (soluble beta glucan) produced by Biotec Pharmamacon ASA (Tromso, Norway) is used in the composition described above. The pharmaceutical composition referred to above may comprise a pharmaceutically acceptable carrier and other ingredients known to enhance and facilitate administration of the drug. The relative amounts of the active ingredient, pharmaceutically acceptable carrier, and any additional ingredients in the pharmaceutical composition of the invention will vary depending on the identity, size, and condition of the subject being treated.

  Such pharmaceutical compositions may include the active ingredient alone, in a form suitable for administration to a subject, or the pharmaceutical composition may comprise the active ingredient and one or more pharmaceutically acceptable substances. A carrier, one or more additional ingredients, or some combination thereof may be included. The active ingredient may be present in the pharmaceutical composition in a form generally well known in the art.

  Formulations of the pharmaceutical compositions described herein can be prepared by any method known or developed below in the pharmacological arts. In general, such preparative methods include the step of bringing into association the active ingredient with the carrier or one or more other auxiliary ingredients and then, when necessary or desirable, the desired single product. Molding or packaging into dose or multi-dose units is included. The sustained-release or sustained-release preparation of the pharmaceutical composition of the present invention can be prepared using conventional techniques.

  In one embodiment, the glucan composition described above can include monoclonal antibodies, or antibodies against cancer or tumor cells, including but not limited to anti-CEA antibodies, anti-CD20 antibodies, anti- CD25 antibody, anti-CD22 antibody, anti-HER2 antibody, anti-tenascin antibody, MoAb M195, dacluzimab, anti-TAG-72 antibody, R24, herceptin, rituximab, 528, IgG, IgM, IgA, C225, epratuzumab, Antibodies to MoAb 3F8 and epidermal growth factor receptor or gangliosides such as GD3 or GD2 are included. In another embodiment, the antibody is a tumor-binding antibody. Preferably, the antibody can activate complement and / or activate cell-mediated cytotoxicity in an antibody-dependent manner. In a further embodiment, the antibody modulates T cell or B cell function.

  In general, antibodies are neuroblastoma, melanoma, non-Hodgkin lymphoma, Epstein-Barr related lymphoma, Hodgkin lymphoma, retinoblastoma, small cell lung cancer, brain tumor, leukemia, squamous cell carcinoma, prostate cancer, renal cell carcinoma, migration It is effective for cancers including epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, colon cancer, liver cancer, gastric cancer, or other gastrointestinal cancers.

  The present invention also provides a method of treating a subject comprising administering to the subject a composition as described above.

  The present invention also provides a composition comprising an effective amount of orally administered (1 → 3), (1 → 6) -β-glucan capable of enhancing the effect of the vaccine. In one embodiment, the vaccine is against cancer or infectious agents such as bacteria, viruses, fungi, or parasites.

  The present invention also provides a composition comprising an effective amount of orally administered (1 → 3), (1 → 6) -β-glucan capable of enhancing host immunity. Host immunity includes, but is not limited to, anti-tumor immune responses.

  In another embodiment, the present invention provides an effective amount of orally administered (1 → 3), (1 → 6) β-glucan that can enhance the action of the substance in preventing tissue rejection. A composition comprising In one embodiment, the tissue is a transplanted tissue, or a transplanted organ, or a graft-versus-host disease host.

  The present invention also provides kits for inhibiting cancer cell growth and / or metastasis. The present invention includes a kit or an administration device comprising a glucan as described herein, and informational material describing the administration of glucan or a composition comprising glucan to a human. The kit or administration device may have a compartment containing the glucan or composition of the invention. As used herein, “information material” is used to convey, but is not limited to, the usefulness of a publication, record, chart, or composition of the invention for its designated use. Any other expression media that can be included.

  The present invention will be further understood by reference to the following experimental description, but those skilled in the art will recognize that the specific detailed experiment is merely exemplary and is as defined by the appended claims. It will be readily understood that it is not meant to limit the invention described in the specification.

Example 1
Yeast β-glucan enhances immune response The entire tumor vaccine can elicit tumor-specific protective immunity in preclinical tumor models. Recent clinical trials using GM-CSF modified, allogeneic or isogenic tumor lines have yielded modest but positive clinical responses. When considering successful vaccines in human medicine, evidence continues to be directed to the importance of antibodies in both inducing and maintaining protective immunity. The continued remission of cancer throughout the completion of the monoclonal antibody strongly suggests that "passive antibody therapy" has induced active immunity. It is hypothesized that when tumor vaccines are opsonized with specific antibodies, their presentation to antigen-presenting cells is enhanced. In the presence of β-glucan, the effectiveness of such a vaccine can be further improved.

  An EL4 isogenic mouse model of lymphoma was used to test the antibody response to the whole tumor vaccine in the presence of β-glucan. When live EL4 tumor cells are transplanted subcutaneously or intravenously into immunocompetent C57B1 / 6 mice, they are rapidly implanted, resulting in death from large tumor masses and metastasis to distant organs. It was. When EL4 tumor cells were transplanted subcutaneously or intravenously in the presence of anti-GD2 antibody 3F8, tumor cell transplantation was reduced. When challenged later with EL4 cells, marginal protective immunity was present. Since β-glucan is known to activate antigen-presenting cells, 3F8 as a tumor vaccine to test whether β-glucan can provide an adjuvant effect to elicit protective immunity. EL4 cells were administered in the presence of

  C57B1 / 6 mice were vaccinated subcutaneously with EL4 lymphoma (as a whole tumor vaccine) in the presence of anti-GD2 antibody 3F8 plus yeast β-glucan. Mouse sera were obtained at 2, 4, and 8 weeks after vaccination. Serum antibodies against surface antigens on EL4 cells were assayed by flow cytometry. Antibodies against whole cell antigen (surface and cytoplasm) were assayed by ELISA using EL4 cells bound to microtiter plates.

  The results from these experiments indicate that (1) 3F8 was necessary to prevent subcutaneous EL4 tumor transplantation, (2) 3F8 enhanced the antibody response against the entire EL4 tumor vaccine , (3) live EL4 tumor vaccine stimulated significantly higher immune response compared to irradiated EL4 tumor vaccine, (4) antibody titer against EL4 tumor increases dose of glucan as adjuvant The optimal dose was 2 mg. (5) The higher the glucan dose, the longer the mice were protected when challenged with intravenous EL4 in a tumor prevention model.

(Example 2)
Phase I study of orally administered yeast β-glucan This phase I study recruited patients with resistant or recurrent metastatic stage 4 neuroblastoma. All patients received 10 mg / m2 / day of anti-GD2 antibody 3F8 for a total of 10 days, while receiving oral yeast β-glucan. The dose of yeast β-glucan was increased in the 3-6 patient age group (10, 20, 40, 80, 100, 120 mg / kg / dose). Eighteen patients were enrolled. There was no dose limiting toxicity (DLT).

  At a dose level of 10 mg / kg, 3 patients were enrolled and treated. These patients completed all four cycles of treatment. Two of these three patients showed a low response. One patient had progressive disease.

  At a dose level of 20 mg / kg, 3 patients were enrolled and treated. One completed all four cycles, had an objective response, and an additional four cycles of treatment were approved by IRB. This patient is currently completing cycle 6. One patient has completed all four cycles and has been assessed for the extent of the disease. One patient has completed 3 cycles of treatment and has developed a human anti-mouse antibody response (HAMA). Assessment of the extent of the disease is undecided.

  At a dose level of 40 mg / kg, 3 patients were enrolled and treated. One completed all four cycles of treatment. Assessment of the extent of disease at the end of 4 cycles revealed disease progression. One patient completed one cycle of treatment. Assessment of the extent of the disease after one cycle revealed disease progression. One patient is currently completing cycle 3.

  At a dose level of 80 mg / kg, 6 patients are registered and treated. One of the patients completed all four cycles of treatment and assessment of the extent of the disease and had a very good partial response (VGPR). One patient completed two cycles. Assessment of the extent of disease after 2 cycles revealed progressive disease. Two patients completed only one cycle of treatment and had progressive disease after one cycle. One patient is undergoing cycle 2 treatment. One patient completed one cycle of treatment. The latter two patients continue the protocol.

  At a dose level of 100 mg / kg, 3 patients were enrolled and treated. There was no dose limiting toxicity. One patient was progressing. One patient achieved a complete remission of bone marrow disease. The latter patient was still too early to assess for response. The latter two patients continue the protocol.

FIG. 5 shows treatment of disseminated human lymphoma in SCID mice using Rituxan and barley or yeast β-glucan. Five million Daudi cells were suspended in 100 μl of normal physiological saline and injected intravenously (iv) into SCID mice. The tumor grew throughout the body, and when the tumor cells infiltrated the spinal cord, the mouse was paralyzed and hindlimb paralyzed. Mice were sacrificed at the start of paralysis or when 10% of the animal's body weight was lost. Treatment started on day 10 after tumor cell injection. 40 μg of rituximab (San Francisco, Calif., Genentech) was intravenously injected twice a week for a total of 8 doses, and 400 μg of glucan was orally administered daily by gavage for 29 days. Mice were weighed weekly and clinical observations were made at least once a day. Mice administered with Rituxan plus barley glucan or Rituxan plus yeast soluble glucan showed a highly significant increase in survival (p <0.05). FIG. 2 shows a ¹ H NMR spectrum of a typical SBG (soluble beta glucan) sample (Biotec Pharmacon ASA, Tromso, Norway). The SBG sample was dissolved in DMSO-d ₆ at a concentration of about 20 mg / ml and a few drops of TFA-d were added. Spectra (excluding 2.7 to 5.5 ppm) were collected at 80 ° C. with a JEOL ECX 400 NMR spectrometer for 2 hours. The chemical shift was based on residual proton resonance from 2.5 ppm DMSO-d ₆ and the spectrum was a corrected baseline. It is a figure which shows the viscosity profile of SBG. Profiles for 2% solutions of SBG at 20 or 30 ° C. at various shear rates were shown. Glycerol (87%) was used as a control solution. It is a figure which shows the (beta) -glucan containing (beta) -1, 3- joint backbone. It is a figure which shows the β-1,3-glucan isolated from the yeast in which β-1,3-linked side chains are tethered to the main chain by β-1,6-linkages.

(References)

Claims (27)

Administering to the subject an anti-tumor antibody that binds to cancer cells in the subject;
Administering to the subject a composition comprising an effective amount of yeast glucan having an average molecular weight of about 16000 to about 17000 daltons and in the range of about 6000 to about 30000 daltons, wherein the glucan exhibits the tumoricidal activity of the antibody. A method for enhancing the tumoricidal activity of an anti-tumor antibody in a subject having cancer.   The method according to claim 1, wherein the antibody is a monoclonal antibody.   3. The method of claim 1 or 2, wherein the antibody is capable of activating complement or activating an antibody dependent cell mediated disorder.   The method according to any one of claims 1 to 3, wherein the antibody recognizes an antigen selected from the group consisting of CD20, CD22, HER-2 / neu, CD25, epidermal growth factor receptor, and ganglioside. .   The method according to claim 4, wherein the ganglioside is GD2 or GD3.   Cancer is neuroblastoma, melanoma, non-Hodgkin lymphoma, Epstein-Barr related lymphoma, Hodgkin lymphoma, retinoblastoma, small cell lung cancer, brain tumor, leukemia, epidermoid cancer, prostate cancer, renal cell cancer, transitional cell carcinoma, 2. The method of claim 1, wherein the method is breast cancer, ovarian cancer, lung cancer, rectal cancer, liver cancer, gastric cancer, or other gastrointestinal cancer.   2. The method of claim 1, wherein the glucan is administered simultaneously with the administration of the antibody or at a different time.   The method of claim 1, wherein the glucan is capable of priming or inducing secretion of cytokines, chemokines, or growth factors by leukocytes. An anti-tumor antibody that binds to cancer cells in a subject and an amount of yeast glucan having an average molecular weight of about 16000 to about 17000 daltons and a range of about 6000 to about 30000 daltons to a subject in need thereof A therapeutic regimen for a subject with cancer that includes a cooperative anti-cancer effect due to the amount of glucan effectively enhances the tumoricidal effect of the antibody.   10. The treatment regimen according to claim 9, wherein the glucan is administered simultaneously with administration of the antibody or at different times.   10. The therapeutic regimen according to claim 9, wherein the antibody is capable of activating complement or activating an antibody dependent cell mediated disorder.   10. The treatment regimen according to claim 9, wherein the antibody is a monoclonal antibody.   13. The therapeutic regimen according to claim 11 or 12, wherein the antibody recognizes an antigen selected from the group consisting of CD20, CD22, HER-2 / neu, CD25, epidermal growth factor receptor, and ganglioside.   10. The treatment regimen of claim 9, wherein the glucan is capable of priming or inducing secretion of cytokines, chemokines, or growth factors by leukocytes. A cancer vaccine composition intended to elicit an anti-tumor immune response in a subject, and an amount of yeast glucan having an average molecular weight of about 16000 to about 17,000 daltons and a range of about 6000 to about 30,000 daltons A therapeutic regimen for a subject with cancer, comprising a cooperative anticancer effect resulting from administration to a subject in need thereof, wherein the amount of glucan effectively enhances the tumoricidal effect of the cancer vaccine composition .   The treatment regimen according to claim 15, wherein the glucan is administered simultaneously with the administration of the cancer vaccine composition or at different times.   16. The treatment regimen of claim 15, wherein the glucan is capable of priming or inducing secretion of cytokines, chemokines, or growth factors by leukocytes.   Use of an effective amount of yeast glucan for preparing an anti-cancer pharmaceutical composition, wherein the average molecular weight of the glucan is about 16000 to about 17000 daltons and the range is about 6000 to about 30000 daltons.   19. The use of claim 18, wherein the anti-cancer agent further comprises a cancer vaccine composition intended to elicit an anti-tumor immune response in the subject.   19. The use according to claim 18, wherein the anti-cancer pharmaceutical composition further comprises an antibody that binds to cancer cells in the subject.   21. Use according to claim 20, wherein the antibody is capable of activating complement or activating an antibody dependent cell mediated disorder.   Use according to claim 20 or 21, wherein the antibody is a monoclonal antibody.   23. Use according to any one of claims 20 to 22, wherein the antibody recognizes an antigen selected from the group consisting of CD20, CD22, HER-2 / neu, CD25, epidermal growth factor receptor, and ganglioside. .   24. Use according to any one of claims 20 to 23, wherein the antibody is administered to the subject simultaneously with glucan administration or at different times.   25. Use according to any one of claims 20 to 24, wherein the antibody binds to cancer cells in a subject with neuroblastoma.   20. Use according to claim 19, wherein the cancer vaccine composition is administered to the subject simultaneously with glucan administration or at different times.   27. Use according to any one of claims 18 to 26, wherein the glucan is capable of priming or inducing secretion of cytokines, chemokines or growth factors by leukocytes.

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